US 2024/0191191 A1 · METHODS FOR INFILTRATING LYMPHOCYTE (TIL) EXPANSION RELATED TO CD39/CD69 SELECTION AND GENE KNOCKOUT IN TILS

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of modified tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from the subject or patient by processing a tumor sample obtained from the subject into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in (a) to obtain a population of CD39/CD69 double negative enriched TILs;
(c) optionally adding the population of CD39/CD69 double negative enriched TILs into a closed system;
(d) performing a first expansion by culturing the population of CD39/CD69 double negative enriched TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-14 days to obtain the second population of TILs, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) harvesting the third population of TILs obtained from step (e), wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (f) to (g) optionally occurs without opening the system;
(h) cryopreserving the infusion bag comprising the harvested third TIL population from step (g) using a cryopreservation process;
(i) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (h) to the subject;
and
(j) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the administering step (i) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

8 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of modified tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from the subject or patient by processing a tumor sample obtained from the subject into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-14 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (e), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(g) cryopreserving the infusion bag comprising the harvested third TIL population from step (f) using a cryopreservation process;
(h) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject;
and
(i) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population TILs at any time prior to the administering step (h) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of modified tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from the subject or patient by processing a tumor sample obtained from the subject into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-14 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (e), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(g) cryopreserving the infusion bag comprising the harvested third TIL population from step (f) using a cryopreservation process;
(h) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject;
and
(i) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population TILs at any time prior to the administering step (h) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of modified tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from the subject or patient by processing a tumor sample obtained from the subject into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-14 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (e), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(g) cryopreserving the infusion bag comprising the harvested third TIL population from step (f) using a cryopreservation process;
(h) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject;
and
(i) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population TILs at any time prior to the administering step (h) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining a first population of TILs from a tumor resected from a subject by processing a tumor sample obtained from the subject into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in (a) to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(c) optionally adding the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs into a closed system;
(d) performing a first expansion by culturing population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) harvesting the third population of TILs obtained from step (e), wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (f) to (g) optionally occurs without opening the system;
(h) cryopreserving the infusion bag comprising the harvested TIL population from step (g) using a cryopreservation process;
(i) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (h) to the subject;
and
(j) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the administering step (i) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

8 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining a first population of TILs from a tumor resected from a subject by processing a tumor sample obtained from the subject into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (e), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(g) cryopreserving the infusion bag comprising the harvested TIL population from step (f) using a cryopreservation process;
(h) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject;
and
(i) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (h) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining a first population of TILs from a tumor resected from a subject by processing a tumor sample obtained from the subject into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (e), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(g) cryopreserving the infusion bag comprising the harvested TIL population from step (f) using a cryopreservation process;
(h) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject;
and
(i) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (h) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining a first population of TILs from a tumor resected from a subject by processing a tumor sample obtained from the subject into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (e), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(g) cryopreserving the infusion bag comprising the harvested TIL population from step (f) using a cryopreservation process;
(h) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject;
and
(i) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (h) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer in the patient or subject,
(b) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in (a) to obtain a population of CD39LO/CD69LO and/or CD39/CD69 enriched TILs;
(c) optionally adding the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs into a closed system;
(d) performing a first expansion by culturing the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) harvesting the third population of TILs obtained from step (e), wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(h) cryopreserving the infusion bag comprising the harvested TIL population from step (g) using a cryopreservation process;
(i) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (h) to the subject;
and
(j) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the administering step (i) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

8 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer in the patient or subject,
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (d), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (e) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(g) cryopreserving the infusion bag comprising the harvested TIL population from step (f) using a cryopreservation process;
(h) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject;
and
(i) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (h) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer in the patient or subject,
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (d), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (e) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(g) cryopreserving the infusion bag comprising the harvested TIL population from step (f) using a cryopreservation process;
(h) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject;
and
(i) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (h) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer in the patient or subject,
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (d), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (e) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(g) cryopreserving the infusion bag comprising the harvested TIL population from step (f) using a cryopreservation process;
(h) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject;
and
(i) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (h) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of modified tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) resecting a tumor from the subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) processing the tumor into multiple tumor fragments;
(c) enzymatically digesting the multiple tumor fragments to obtain the first population of TILs;
(d) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in (c) to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(e) optionally adding the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs into a closed system;
(f) performing a first expansion by culturing the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (f) to step (g) optionally occurs without opening the system;
(h) harvesting the third population of TILs obtained from step (g), wherein the transition from step (g) to step (h) optionally occurs without opening the system;
(i) transferring the harvested third TIL population from step (h) to an infusion bag, wherein the transfer from step (h) to (i) optionally occurs without opening the system;
(j) cryopreserving the infusion bag comprising the harvested TIL population from step (i) using a cryopreservation process;
(k) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject or patient with the cancer;
and
(l) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (k) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

10 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of modified tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) resecting a tumor from the subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) processing the tumor into multiple tumor fragments;
(c) enzymatically digesting the multiple tumor fragments to obtain the first population of TILs;
(d) optionally adding the tumor fragments or tumor digest into a closed system;
(e) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) harvesting the third population of TILs obtained from step (f), wherein the transition from step (f) to step (g) optionally occurs without opening the system;
(h) transferring the harvested third TIL population from step (g) to an infusion bag, wherein the transfer from step (g) to (h) optionally occurs without opening the system;
(i) cryopreserving the infusion bag comprising the harvested TIL population from step (h) using a cryopreservation process;
(l) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (h) to the subject or patient with the cancer;
and
(k) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (1) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

9 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of modified tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) resecting a tumor from the subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) processing the tumor into multiple tumor fragments;
(c) enzymatically digesting the multiple tumor fragments to obtain the first population of TILs;
(d) optionally adding the tumor fragments or tumor digest into a closed system;
(e) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(f) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) harvesting the third population of TILs obtained from step (f), wherein the transition from step (f) to step (g) optionally occurs without opening the system;
(h) transferring the harvested third TIL population from step (g) to an infusion bag, wherein the transfer from step (g) to (h) optionally occurs without opening the system;
(i) cryopreserving the infusion bag comprising the harvested TIL population from step (h) using a cryopreservation process;
(j) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (h) to the subject or patient with the cancer;
and
(k) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (1) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

9 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of modified tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) resecting a tumor from the subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) processing the tumor into multiple tumor fragments;
(c) enzymatically digesting the multiple tumor fragments to obtain the first population of TILs;
(d) optionally adding the tumor fragments or tumor digest into a closed system;
(e) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-14 days to obtain the second population of TILs, and wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) harvesting the third population of TILs obtained from step (f), wherein the transition from step (f) to step (g) optionally occurs without opening the system;
(h) transferring the harvested third TIL population from step (g) to an infusion bag, wherein the transfer from step (g) to (h) optionally occurs without opening the system;
(i) cryopreserving the infusion bag comprising the harvested TIL population from step (h) using a cryopreservation process;
(j) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (h) to the subject or patient with the cancer;
and
(k) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (1) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

9 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the subject or patient;
(b) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in (a) to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(c) contacting the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs with a first cell culture medium;
(d) performing an initial expansion (or priming first expansion) of the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs in the first cell culture medium to obtain a second population of TILs, wherein the first cell culture medium comprises IL-2, optionally OKT-3 (anti-CD3 antibody), and optionally antigen presenting cells (APCs), where the priming first expansion occurs for a period of 1 to 8 days;
(e) performing a rapid second expansion of the second population of TILs in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), and APCs;
and wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(f) harvesting the third population of TILs;
(g) administering a therapeutically effective portion of the third population of TILs to the subject or patient with the cancer;
and
(h) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the administering step (g) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

8 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the subject or patient;
(b) performing an initial expansion (or priming first expansion) by culturing the first population of TILs in a cell culture medium comprising IL-2, optionally OKT-3 (anti-CD3 antibody), optionally antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the priming first expansion is performed for about 1-8 days to obtain the second population of TILs, and wherein the transition from step (a) to step (b) optionally occurs without opening the system;
(c) performing a rapid second expansion of the second population of TILs in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), and APCs;
and wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(d) harvesting the third population of TILs;
(e) administering a therapeutically effective portion of the third population of TILs to the subject or patient with the cancer;
and
(f) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (e) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

6 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the subject or patient;
(b) performing an initial expansion (or priming first expansion) of the first population of TILs in a first cell culture medium to obtain a second population of TILs, wherein the first cell culture medium comprises IL-2, optionally OKT-3 (anti-CD3 antibody), and optionally antigen presenting cells (APCs), where the priming first expansion occurs for a period of 1 to 8 days;
(c) performing a rapid second expansion in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(d) harvesting the third population of TILs;
(e) administering a therapeutically effective portion of the third population of TILs to the subject or patient with the cancer;
and
(f) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (e) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

5 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the subject or patient;
(b) performing an initial expansion (or priming first expansion) by culturing the first population of TILs in a first cell culture medium comprising IL-2, optionally OKT-3 (anti-CD3 antibody), optionally antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the priming first expansion is performed for about 1-8 days to obtain the second population of TILs, and wherein the transition from step (a) to step (b) optionally occurs without opening the system;
(c) performing a rapid second expansion in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(d) harvesting the third population of TILs;
(e) administering a therapeutically effective portion of the third population of TILs to the subject or patient with the cancer;
and
(f) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (e) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

5 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) resecting a tumor from the cancer in the subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) fragmenting the tumor into tumor fragments or processing th tumor into a tumor digest;
(c) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs of the tumor fragments to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(c) contacting the tumor fragments with a first cell culture medium;
(d) performing an initial expansion (or priming first expansion) of the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs in the first cell culture medium to obtain a second population of TILs, wherein the first cell culture medium comprises IL-2, optionally OKT-3 (anti-CD3 antibody), and optionally antigen presenting cells (APCs), where the priming first expansion occurs for a period of 1 to 8 days;
(e) performing a rapid second expansion of the second population of TILs in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), and APCs;
and wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(f) harvesting the third population of TILs;
(g) administering a therapeutically effective portion of the third population of TILs to the subject or patient with the cancer;
and
(h) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the administering step (g) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

9 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) resecting a tumor from the cancer in the subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) fragmenting the tumor into tumor fragments or processing th tumor into a tumor digest;
(c) performing an initial expansion (or priming first expansion) by culturing the first population of TILs in a first cell culture medium comprising IL-2, optionally OKT-3 (anti-CD3 antibody), optionally antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the priming first expansion is performed for about 1-8 days to obtain the second population of TILs, and wherein the transition from step (a) to step (b) optionally occurs without opening the system;
(d) performing a rapid second expansion of the second population of TILs in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), and APCs;
and wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(e) harvesting the third population of TILs;
(f) administering a therapeutically effective portion of the third population of TILs to the subject or patient with the cancer;
and
(g) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (f) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) resecting a tumor from the cancer in the subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) fragmenting the tumor into tumor fragments or processing th tumor into a tumor digest;
(c) performing an initial expansion (or priming first expansion) of the first population of TILs in a first cell culture medium to obtain a second population of TILs, wherein the first cell culture medium comprises IL-2, optionally OKT-3 (anti-CD3 antibody), and optionally antigen presenting cells (APCs), where the priming first expansion occurs for a period of 1 to 8 days;
(d) performing a rapid second expansion in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(e) harvesting the third population of TILs;
(f) administering a therapeutically effective portion of the third population of TILs to the subject or patient with the cancer;
and
(g) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (f) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

6 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) resecting a tumor from the cancer in the subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) fragmenting the tumor into tumor fragments or processing th tumor into a tumor digest;
(c) performing an initial expansion (or priming first expansion) by culturing the first population of TILs in a first cell culture medium comprising IL-2, optionally OKT-3 (anti-CD3 antibody), optionally antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the priming first expansion is performed for about 1-8 days to obtain the second population of TILs, and wherein the transition from step (a) to step (b) optionally occurs without opening the system;
(d) performing a rapid second expansion in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(e) harvesting the third population of TILs;
(f) administering a therapeutically effective portion of the third population of TILs to the subject or patient with the cancer;
and
(g) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the administering step (f) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

6 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer in the patient or subject,
(b) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in (a) to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(c) performing a priming first expansion by culturing the CD39LO/CD69LO and/or CD39/CD69 double negative enriched TIL population in a first cell culture medium comprising IL-2, OKT-3, and antigen presenting cells (APCs) to produce a second population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(d) optionally restimulating the second population of TILs with OKT-3;
(e) genetically modifying the second population of TILs to produce a modified second population of TILs, wherein the modified second population of TILs comprises a genetic modification that reduces the expression of CD39 and CD69 such that the second population comprises CD39LO/CD69LO and/or CD39/CD69 double negative TILs;
(f) performing a rapid second expansion by culturing the modified second population of TILs in a second culture medium comprising IL-2, OKT-3, and APCs, to produce a third population of TILs, wherein the rapid second expansion is performed for a second period of about 14 days or less to obtain the therapeutic population of TILs, wherein the third population of TILs is a therapeutic population of TILs comprising the genetic modification that reduces the expression of CD39 and CD69 such that the third population comprises CD39LO/CD69LO and/or CD39/CD69 double negative TILs;
(g) harvesting the third population of TILs;
and
(h) administering a therapeutically effective portion of the third population of TILs to the subject or patient with the cancer.

6 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer in the patient or subject,
(b) performing a priming first expansion by culturing the first population of TILs in a first cell culture medium comprising IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(c) optionally restimulating the second population of TILs with OKT-3;
(d) genetically modifying the second population of TILs to produce a modified second population of TILs, wherein the modified second population of TILs comprises a genetic modification that reduces the expression of CD39 and CD69 such that the second population comprises CD39LO/CD69LO and/or CD39/CD69 double negative TILs;
(e) performing a rapid second expansion by culturing the modified second population of TILs in a second culture medium comprising IL-2, OKT-3, and APCs, to produce a third population of TILs, wherein the rapid second expansion is performed for a second period of about 14 days or less to obtain the therapeutic population of TILs, wherein the third population of TILs is a therapeutic population of TILs comprising the genetic modification that reduces the expression of CD39 and CD69 such that the third population comprises CD39LO/CD69LO and/or CD39/CD69 double negative TILs;
(f) harvesting the third population of TILs;
and
(g) administering a therapeutically effective portion of the third population of TILs to the subject or patient with the cancer.

5 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer in the patient or subject,
(b) performing a priming first expansion by culturing the first population of TILs in a first cell culture medium comprising IL-2, OKT-3, and antigen presenting cells (APCs) to produce a second population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(c) optionally restimulating the second population of TILs with OKT-3;
(d) genetically modifying the second population of TILs to produce a modified second population of TILs, wherein the modified second population of TILs comprises a genetic modification that reduces the expression of CD39 and CD69 such that the second population comprises CD39LO/CD69LO and/or CD39/CD69 double negative TILs;
(e) performing a rapid second expansion by culturing the modified second population of TILs in a second culture medium comprising IL-2, OKT-3, APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid second expansion is performed for a second period of about 14 days or less to obtain the therapeutic population of TILs, wherein the third population of TILs is a therapeutic population of TILs comprising the genetic modification that reduces the expression of CD39 and CD69 such that the third population comprises CD39LO/CD69LO and/or CD39/CD69 double negative TILs;
(f) harvesting the third population of TILs;
and
(g) administering a therapeutically effective portion of the third population of TILs to the subject or patient with the cancer.

5 more elements

A method of treating a cancer in a patient or subject in need thereof comprising administering a population of tumor infiltrating lymphocytes (TILs), the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer in the patient or subject,
(b) performing a priming first expansion by culturing the first population of TILs in a first cell culture medium comprising IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(c) optionally restimulating the second population of TILs with OKT-3;
(d) genetically modifying the second population of TILs to produce a modified second population of TILs, wherein the modified second population of TILs comprises a genetic modification that reduces the expression of CD39 and CD69 such that the second population comprises CD39LO/CD69LO and/or CD39/CD69 double negative TILs;
(e) performing a rapid second expansion by culturing the modified second population of TILs in a second culture medium comprising IL-2, OKT-3, APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid second expansion is performed for a second period of about 14 days or less to obtain the therapeutic population of TILs, wherein the third population of TILs is a therapeutic population of TILs comprising the genetic modification that reduces the expression of CD39 and CD69 such that the third population comprises CD39LO/CD69LO and/or CD39/CD69 double negative TILs;
(f) harvesting the third population of TILs;
(g) administering a therapeutically effective portion of the third population of TILs to the subject or patient with the cancer.

4 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from a cancer in a subject by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in step (a) to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(c) performing a priming first expansion by culturing the CD39LO/CD69LO and/or CD39/CD69 double negative enriched TIL population in a first cell culture medium comprising IL-2, OKT-3, and antigen presenting cells (APCs) to produce a second population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 7/8 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(d) performing a rapid second expansion by culturing the second population of TILs in a second culture medium comprising IL-2, OKT-3, and APCs, to produce a third population of TILs, wherein the number of APCs added in the rapid second expansion is at least twice the number of APCs added in step (b), wherein the rapid second expansion is performed for a second period of about 1 to 11 days to obtain the therapeutic population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the rapid second expansion is performed in a container comprising a second gas-permeable surface area;
(e) harvesting the therapeutic population of TILs obtained from step (d);
(f) transferring the harvested TIL population from step (e) to an infusion bag;
and
(g) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or second population of TILs and/or third population of TILs at any time prior to the harvesting step (e) such that the therapeutic population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

5 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from a cancer in a subject by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) performing a priming first expansion by culturing the first population of TILs in a first cell culture medium comprising IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 7/8 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(c) performing a rapid second expansion by culturing the second population of TILs in a second culture medium comprising IL-2, OKT-3, and APCs, to produce a third population of TILs, wherein the number of APCs added in the rapid second expansion is at least twice the number of APCs added in step (b), wherein the rapid second expansion is performed for a second period of about 1 to 11 days to obtain the therapeutic population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the rapid second expansion is performed in a container comprising a second gas-permeable surface area;
(d) harvesting the therapeutic population of TILs obtained from step (d);
(e) transferring the harvested TIL population from step (e) to an infusion bag;
and
(f) optionally genetically modifying the first population of TILs, the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (e) such that the therapeutic population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

4 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from a cancer in a subject by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) performing a priming first expansion by culturing the first population of TILs in a first cell culture medium comprising IL-2, OKT-3, and antigen presenting cells (APCs) to produce a second population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 7/8 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(c) performing a rapid second expansion by culturing the second population of TILs in a second culture medium comprising IL-2, OKT-3, APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the number of APCs added in the rapid second expansion is at least twice the number of APCs added in step (b), wherein the rapid second expansion is performed for a second period of about 1 to 11 days to obtain the therapeutic population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the rapid second expansion is performed in a container comprising a second gas-permeable surface area;
(d) harvesting the therapeutic population of TILs obtained from step (d);
(e) transferring the harvested TIL population from step (e) to an infusion bag;
and
(f) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (e) such that the therapeutic population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

4 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from a cancer in a subject by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) performing a priming first expansion by culturing the first population of TILs in a first cell culture medium comprising IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 7/8 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(c) performing a rapid second expansion by culturing the second population of TILs in a second culture medium comprising IL-2, OKT-3, APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the number of APCs added in the rapid second expansion is at least twice the number of APCs added in step (b), wherein the rapid second expansion is performed for a second period of about 1 to 11 days to obtain the therapeutic population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the rapid second expansion is performed in a container comprising a second gas-permeable surface area;
(d) harvesting the therapeutic population of TILs obtained from step (d);
(e) transferring the harvested TIL population from step (e) to an infusion bag;
and
(f) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (e) such that the therapeutic population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

4 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from a cancer in a subject or patient by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in (a) to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(c) optionally adding the population of CD39LO/CD69LO and/or CD39/CD69 double negative TILs into a closed system;
(d) performing a first expansion by culturing the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-14 days to obtain the second population of TILs, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) harvesting the third population of TILs obtained from step (e), wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (f) to (g) optionally occurs without opening the system;
and
(h) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or the second population of TILs and/or the third population of TILS at any time prior to the harvesting step (f) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

6 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from a cancer in a subject or patient by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-14 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (d), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (e) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
and
(g) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (f) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

5 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from a cancer in a subject or patient by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-14 days to obtain the second population of TILs, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (d), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (e) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
and
(g) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (f) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

5 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from a cancer in a subject or patient by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-14 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (d), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (e) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
and
(g) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (f) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

5 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining a first population of TILs from a tumor resected from a cancer in a subject by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
b) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in (a) to obtain a population of (i) CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(c) optionally adding the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs into a closed system;
(d) performing a first expansion by culturing population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) harvesting the third population of TILs obtained from step (e), wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (f) to (g) optionally occurs without opening the system;
and
(h) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (f) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

6 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining a first population of TILs from a tumor resected from a subject by processing a tumor sample obtained from the subject into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (e), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(g) cryopreserving the infusion bag comprising the harvested TIL population from step (f) using a cryopreservation process;
(h) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject;
and
(i) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the administering step (h) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining a first population of TILs from a tumor resected from a subject by processing a tumor sample obtained from the subject into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (e), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(g) cryopreserving the infusion bag comprising the harvested TIL population from step (f) using a cryopreservation process;
(h) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject;
and
(i) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the administering step (h) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining a first population of TILs from a tumor resected from a subject by processing a tumor sample obtained from the subject into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (e), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
(g) cryopreserving the infusion bag comprising the harvested TIL population from step (f) using a cryopreservation process;
(h) administering a therapeutically effective dosage of the third population of TILs from the infusion bag in step (g) to the subject;
and
(i) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the administering step (h) such that the administered third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from a cancer in a patient or subject,
(b) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in (a) to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative, enriched TILs;
(c) optionally adding the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs into a closed system;
(d) performing a first expansion by culturing the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) harvesting the third population of TILs obtained from step (e), wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) transferring the harvested third TIL population from step (f) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
and
(h) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (f) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

6 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer in the patient or subject,
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (d), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (e) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
and
(g) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (e) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

5 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer in the patient or subject,
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (d), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (e) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
and
(g) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (e) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

5 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer in the patient or subject,
(b) optionally adding the tumor fragments or tumor digest into a closed system;
(c) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (b) to step (c) optionally occurs without opening the system;
(d) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (c) to step (d) optionally occurs without opening the system;
(e) harvesting the third population of TILs obtained from step (d), wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) transferring the harvested third TIL population from step (e) to an infusion bag, wherein the transfer from step (e) to (f) optionally occurs without opening the system;
and
(g) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (e) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

5 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) to a therapeutic population of TILs, the method comprising the steps of:

(a) resecting a tumor from a cancer in subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) processing the tumor into multiple tumor fragments or into a tumor digest;
(c) enzymatically digesting the multiple tumor fragments to obtain the first population of TILs;
(d) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in (c) to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(e) optionally adding the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs into a closed system;
(f) performing a first expansion by culturing the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (f) to step (g) optionally occurs without opening the system;
(h) harvesting the third population of TILs obtained from step (g), wherein the transition from step (g) to step (h) optionally occurs without opening the system;
(i) transferring the harvested third TIL population from step (h) to an infusion bag, wherein the transfer from step (h) to (i) optionally occurs without opening the system;
and
(j) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (h) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

8 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) to a therapeutic population of TILs, the method comprising the steps of:

(a) resecting a tumor from a cancer in subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) processing the tumor into multiple tumor fragments or into a tumor digest;
(c) enzymatically digesting the multiple tumor fragments to obtain the first population of TILs;
(d) optionally adding the tumor fragments or tumor digest into a closed system;
(e) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) harvesting the third population of TILs obtained from step (f), wherein the transition from step (f) to step (g) optionally occurs without opening the system;
(h) transferring the harvested third TIL population from step (g) to an infusion bag, wherein the transfer from step (g) to (h) optionally occurs without opening the system;
and
(i) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (g) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) to a therapeutic population of TILs, the method comprising the steps of:

(a) resecting a tumor from a cancer in subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) processing the tumor into multiple tumor fragments or into a tumor digest;
(c) enzymatically digesting the multiple tumor fragments to obtain the first population of TILs;
(d) optionally adding the tumor fragments or tumor digest into a closed system;
(e) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(f) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) harvesting the third population of TILs obtained from step (f), wherein the transition from step (f) to step (g) optionally occurs without opening the system;
(h) transferring the harvested third TIL population from step (g) to an infusion bag, wherein the transfer from step (g) to (h) optionally occurs without opening the system;
and
(i) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (g) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) to a therapeutic population of TILs, the method comprising the steps of:

(a) resecting a tumor from a cancer in subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) processing the tumor into multiple tumor fragments or into a tumor digest;
(c) enzymatically digesting the multiple tumor fragments to obtain the first population of TILs;
(d) optionally adding the tumor fragments or tumor digest into a closed system;
(e) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for about 3-11 days to obtain the second population of TILs, and wherein the transition from step (d) to step (e) optionally occurs without opening the system;
(f) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the second expansion is performed for about 7-11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is optionally performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (e) to step (f) optionally occurs without opening the system;
(g) harvesting the third population of TILs obtained from step (f), wherein the transition from step (f) to step (g) optionally occurs without opening the system;
(h) transferring the harvested third TIL population from step (g) to an infusion bag, wherein the transfer from step (g) to (h) optionally occurs without opening the system;
and
(i) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (g) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from a cancer in the subject or patient;
(b) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in (a) to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(c) contacting the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs with a first cell culture medium;
(d) performing an initial expansion (or priming first expansion) of the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs in the first cell culture medium to obtain a second population of TILs, wherein the first cell culture medium comprises IL-2, optionally OKT-3 (anti-CD3 antibody), and optionally antigen presenting cells (APCs), where the priming first expansion occurs for a period of 1 to 8 days;
(e) performing a rapid second expansion of the second population of TILs in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), and APCs;
and wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(f) harvesting the third population of TILs;
and
(g) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (f) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

7 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from a cancer in the subject or patient;
(b) performing an initial expansion (or priming first expansion) by culturing the first population of TILs in a cell culture medium comprising IL-2, optionally OKT-3 (anti-CD3 antibody), optionally antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the priming first expansion is performed for about 1-8 days to obtain the second population of TILs, and wherein the transition from step (a) to step (b) optionally occurs without opening the system;
(c) performing a rapid second expansion of the second population of TILs in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), and APCs;
and wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(d) harvesting the third population of TILs;
and
(e) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (d) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

5 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from a cancer in the subject or patient;
(b) performing an initial expansion (or priming first expansion) of the first population of TILs in a first cell culture medium to obtain a second population of TILs, wherein the first cell culture medium comprises IL-2, optionally OKT-3 (anti-CD3 antibody), and optionally antigen presenting cells (APCs), where the priming first expansion occurs for a period of 1 to 8 days;
(c) performing a rapid second expansion in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(d) harvesting the third population of TILs;
and
(e) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (d) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

4 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from a cancer in the subject or patient;
(b) performing an initial expansion (or priming first expansion) by culturing the first population of TILs in a cell culture medium comprising IL-2, optionally OKT-3 (anti-CD3 antibody), optionally antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the priming first expansion is performed for about 1-8 days to obtain the second population of TILs, and wherein the transition from step (a) to step (b) optionally occurs without opening the system;
(c) performing a rapid second expansion in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(d) harvesting the third population of TILs;
and
(e) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (d) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

4 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

a) resecting a tumor from the cancer in the subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) fragmenting the tumor into tumor fragments or into a tumor digest;
(c) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in the tumor fragments or tumor digest to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(d) contacting the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs with a first cell culture medium;
(e) performing an initial expansion (or priming first expansion) of the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs in the first cell culture medium to obtain a second population of TILs, wherein the first cell culture medium comprises IL-2, optionally OKT-3 (anti-CD3 antibody), and optionally antigen presenting cells (APCs), where the priming first expansion occurs for a period of 1 to 8 days;
(f) performing a rapid second expansion of the second population of TILs in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), and APCs;
and wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(g) harvesting the third population of TILs;
and
(h) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting (f) such that the harvested third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

8 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

a) resecting a tumor from the cancer in the subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) fragmenting the tumor into tumor fragments or into a tumor digest;
(c) performing an initial expansion (or priming first expansion) by culturing the first population of TILs in a first cell culture medium comprising IL-2, optionally OKT-3 (anti-CD3 antibody), optionally antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the priming first expansion is performed for about 1-8 days to obtain the second population of TILs, and wherein the transition from step (a) to step (b) optionally occurs without opening the system;
(d) performing a rapid second expansion of the second population of TILs in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), and APCs;
and wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(e) harvesting the third population of TILs;
and
(f) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting (e) such that the harvested third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

6 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

a) resecting a tumor from the cancer in the subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) fragmenting the tumor into tumor fragments or into a tumor digest;
(c) performing an initial expansion (or priming first expansion) of the first population of TILs in a first cell culture medium to obtain a second population of TILs, wherein the first cell culture medium comprises IL-2, optionally OKT-3 (anti-CD3 antibody), and optionally antigen presenting cells (APCs), where the priming first expansion occurs for a period of 1 to 8 days;
(d) performing a rapid second expansion in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(e) harvesting the third population of TILs;
and
(f) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting (e) such that the harvested third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

5 more elements

A method of expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs, the method comprising the steps of:

a) resecting a tumor from the cancer in the subject or patient, the tumor comprising a first population of TILs, optionally from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from the cancer;
(b) fragmenting the tumor into tumor fragments or into a tumor digest;
(c) performing an initial expansion (or priming first expansion) by culturing the first population of TILs in a first cell culture medium comprising IL-2, optionally OKT-3 (anti-CD3 antibody), optionally antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the first expansion is optionally performed in a closed container providing a first gas-permeable surface area, wherein the priming first expansion is performed for about 1-8 days to obtain the second population of TILs, and wherein the transition from step (a) to step (b) optionally occurs without opening the system;
(d) performing a rapid second expansion in a second cell culture medium to obtain a third population of TILs;
wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid expansion is performed over a period of 14 days or less, optionally the rapid second expansion can proceed for 1 day, 2 days, 3 days, 4, days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days after initiation of the rapid second expansion;
(e) harvesting the third population of TILs;
and
(f) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting (e) such that the harvested third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

5 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from a cancer in a subject by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in step (a) to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(c) performing a priming first expansion by culturing the population of CD39/CD69 double negative enriched TILs in a cell culture medium comprising IL-2, optionally OKT-3, and optionally comprising antigen presenting cells (APCs), to produce a second population of TILs, wherein the priming first expansion is performed for a first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(d) performing a rapid second expansion by contacting the second population of TILs with a second cell culture medium comprising IL-2, OKT-3, and APCs, to produce a third population of TILs, wherein the rapid second expansion is performed for a second period of about 1 to 11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs;
(e) harvesting the therapeutic population of TILs obtained from step (c);
and
(f) optionally genetically modifying the population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (e) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

4 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from a cancer in a subject by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) performing a priming first expansion by culturing the first population of TILs in a first cell culture medium comprising IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(c) performing a rapid second expansion by contacting the second population of TILs with a second cell culture medium comprising IL-2, OKT-3, and APCs, to produce a third population of TILs, wherein the rapid second expansion is performed for a second period of about 1 to 11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs;
(d) harvesting the therapeutic population of TILs obtained from step (c);
and
(e) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (d) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

3 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from a cancer in a subject by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) performing a priming first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2, optionally OKT-3, and optionally comprising antigen presenting cells (APCs), to produce a second population of TILs, wherein the priming first expansion is performed for a first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(c) performing a rapid second expansion by contacting the second population of TILs with a cell culture medium comprising IL-2, OKT-3, APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid second expansion is performed for a second period of about 1 to 11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs;
(d) harvesting the therapeutic population of TILs obtained from step (c);
and
(e) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (d) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

3 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) obtaining and/or receiving a first population of TILs from a tumor resected from a cancer in a subject by processing a tumor sample obtained from the tumor into multiple tumor fragments or processing a tumor sample obtained from the subject into a tumor digest;
(b) performing a priming first expansion by culturing the first population of TILs in a first cell culture medium comprising IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(c) performing a rapid second expansion by contacting the second population of TILs with a cell culture medium comprising IL-2, OKT-3, APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid second expansion is performed for a second period of about 1 to 11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs;
(d) harvesting the therapeutic population of TILs obtained from step (c);
and
(e) optionally genetically modifying the first population of TILs and/or the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (d) such that the third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

3 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from a cancer in a patient or subject,
(b) selecting CD39LO/CD69LO and/or CD39/CD69 double negative TILs from the first population of TILs in (a) to obtain a population of CD39LO/CD69LO and/or CD39/CD69 double negative enriched TILs;
(c) performing a priming first expansion by culturing the CD39LO/CD69LO and/or CD39/CD69 double negative enriched TIL population in a first cell culture medium comprising IL-2, OKT-3, and antigen presenting cells (APCs) to produce a second population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(d) optionally restimulating the second population of TILs with OKT-3;
(e) genetically modifying the second population of TILs to produce a modified second population of TILs, wherein the modified second population of TILs comprises a genetic modification that reduces the expression of CD39 and CD69;
(f) performing a rapid second expansion by culturing the modified second population of TILs in a second culture medium comprising IL-2, OKT-3, and APCs, to produce a third population of TILs, wherein the rapid second expansion is performed for a second period of about 14 days or less to obtain the therapeutic population of TILs, wherein the third population of TILs is a therapeutic population of TILs comprises the genetic modification that reduces the expression of CD39 and CD69;
and
(g) harvesting the third population of TILs.

5 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from a cancer in a patient or subject,
(b) performing a priming first expansion by culturing the first population of TILs in a first cell culture medium comprising IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(c) optionally restimulating the second population of TILs with OKT-3;
(d) genetically modifying the second population of TILs to produce a modified second population of TILs, wherein the modified second population of TILs comprises a genetic modification that reduces the expression of CD39 and CD69;
(e) performing a rapid second expansion by culturing the modified second population of TILs in a second culture medium comprising IL-2, OKT-3, and APCs, to produce a third population of TILs, wherein the rapid second expansion is performed for a second period of about 14 days or less to obtain the therapeutic population of TILs, wherein the third population of TILs is a therapeutic population of TILs comprises the genetic modification that reduces the expression of CD39 and CD69;
and
(f) harvesting the third population of TILs.

4 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from a cancer in a patient or subject,
(b) performing a priming first expansion by culturing the first population of TILs in a first cell culture medium comprising IL-2, OKT-3, and antigen presenting cells (APCs) to produce a second population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(c) optionally restimulating the second population of TILs with OKT-3;
(d) genetically modifying the second population of TILs to produce a modified second population of TILs, wherein the modified second population of TILs comprises a genetic modification that reduces the expression of CD39 and CD69;
(e) performing a rapid second expansion by culturing the modified second population of TILs in a second culture medium comprising IL-2, OKT-3, APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid second expansion is performed for a second period of about 14 days or less to obtain the therapeutic population of TILs, wherein the third population of TILs is a therapeutic population of TILs comprises the genetic modification that reduces the expression of CD39 and CD69;
and
(f) harvesting the third population of TILs.

4 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) obtaining and/or receiving a first population of TILs from surgical resection, needle biopsy, core biopsy, small biopsy, or other means for obtaining a sample that contains a mixture of tumor and TIL cells from a cancer in a patient or subject,
(b) performing a priming first expansion by culturing the first population of TILs in a first cell culture medium comprising IL-2, OKT-3, antigen presenting cells (APCs), and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a second population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the priming first expansion is performed in a container comprising a first gas-permeable surface area, wherein the priming first expansion is performed for first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(c) optionally restimulating the second population of TILs with OKT-3;
(d) genetically modifying the second population of TILs to produce a modified second population of TILs, wherein the modified second population of TILs comprises a genetic modification that reduces the expression of CD39 and CD69;
(e) performing a rapid second expansion by culturing the modified second population of TILs in a second culture medium comprising IL-2, OKT-3, APCs, and a protein kinase B (AKT) inhibitor, optionally wherein the AKT inhibitor is selected from the group consisting of ipatasertib, GSK690693, GSK2141795, GSK2110183, AZD5363, GDC-0068, AT7867, CCT128930, MK-2206, BAY 1125976, Perifosine, Oridonin, Herbacetin, Tehranolide, Isoliquiritigenin, Scutellarin, and Honokiol, to produce a third population of TILs that is a CD39LO/CD69LO and/or CD39/CD69 double negative enriched population of TILs, wherein the rapid second expansion is performed for a second period of about 14 days or less to obtain the therapeutic population of TILs, wherein the third population of TILs is a therapeutic population of TILs comprises the genetic modification that reduces the expression of CD39 and CD69;
and
(f) harvesting the third population of TILs.

4 more elements

A method for expanding tumor infiltrating lymphocytes (TILs) into a therapeutic population of TILs comprising:

(a) performing a priming first expansion by culturing a first population of CD39/CD69 double negative and/or CD39LO/CD69LO enriched TILs in a cell culture medium comprising IL-2, optionally OKT-3, and optionally comprising antigen presenting cells (APCs), to produce a second population of TILs, wherein the priming first expansion is performed for a first period of about 1 to 11 days to obtain the second population of TILs, wherein the second population of TILs is greater in number than the first population of TILs;
(b) performing a rapid second expansion by contacting the second population of TILs with a second cell culture medium comprising IL-2, OKT-3, and APCs, to produce a third population of TILs, wherein the rapid second expansion is performed for a second period of about 1 to 11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs;
and
(c) harvesting the third population of TILs obtained from step (b).
(d) genetically modifying the population of CD39/CD69 double negative and/or CD39LO/CD69LO enriched TILs, the second population of TILs and/or the third population of TILs at any time prior to the harvesting step (c) such that the harvested third population of TILs comprises genetically modified TILs comprising a genetic modification that reduces the expression of CD39 and CD69.

2 more elements

Methods for infiltrating lymphocyte (til) expansion related to cd39/cd69 selection and gene knockout in tils

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Methods for infiltrating lymphocyte (til) expansion related to cd39/cd69 selection and gene knockout in tils

Life of this patent

Abstract

Claims135