Engineered cells for therapy
AssigneeShoreline Biosciences, Inc.
InventorsJohn Anthony ZURIS, Carrie Marie MARGULIES
Life of this patent
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Abstract
Strategies, systems, compositions, and methods for genetically modifying cells to include one or more loss-of-function modifications and/or to include one or more gain-of-function modifications, as well as modified cells (and compositions of such cells) that include one or more loss-of-function modifications and/or that include one or more gain-of-function modifications, are described. In certain aspects, such modified cells include at least one gain-of-function modification within a coding region of an essential gene.
Claims63
A Natural Killer (NK) cell comprising:
- (a) one or more genomic edits that results in loss of function of one or more genes, wherein the one or more genes encode adenosine A2a receptor (ADORA2A), β-2 microglobulin (B2M), class II major histocompatibility complex transactivator (CIITA), cytokine inducible SH2 containing protein (CISH), two or more human leukocyte antigen (HLA) class II histocompatibility antigen alpha chain genes, two or more HLA class II histocompatibility antigen beta chain genes, natural killer group 2 member A receptor (NKG2A), programmed cell death protein 1 (PD-1), T cell immunoreceptor with Ig and ITIM domains (TIGIT), an agonist of the TGF beta signaling pathway (e.g., transforming growth factor beta receptor II (TGFβRII)), or any combination of two or more thereof;
- and
- (b) a genome comprising a first exogenous coding sequence for FcγRIII (CD16) or variant thereof and a second exogenous coding sequence for a membrane bound interleukin 15 (mbIL-15), wherein the first exogenous coding sequence and second exogenous coding sequence are in frame with and downstream (3′) of a coding sequence of an essential gene, and wherein at least part of the essential gene comprises an exogenous coding sequence.
The NK cell of any one of the preceding claims, wherein the first exogenous coding sequence is or comprises SEQ ID NO:
- 166.
The NK cell of any one of the preceding claims, wherein the second exogenous coding sequence is or comprises SEQ ID NO:
- 172.
The NK cell of any one of the preceding claims, wherein the CD16 is or comprises the amino acid sequence of SEQ ID NO:
- 184.
The NK cell of any one of the preceding claims, wherein the mbIL-15 comprises an IL-15, a linker, a sushi domain, and an IL-15Rα.
The NK cell of any one of the preceding claims, wherein the NK cell is an induced pluripotent stem cell (iPSC)-derived NK (INK) cell.
The NK cell of any one of the preceding claims, wherein the essential gene encodes a gene product that is required for survival and/or proliferation of the cell.
The NK cell of any one of the preceding claims, wherein the essential gene is a housekeeping gene, e.g., a gene listed in Table 3.
The NK cell of any one of the preceding claims, wherein the essential gene encodes glyceraldehyde 3-phosphate dehydrogenase (GAPDH).
The NK cell of any one of the preceding claims, wherein the NK cell comprises:
- (i) a genomic edit that results in loss of function of CISH;
- and
- (ii) a genomic edit that results in loss of function of TGFβRII.
The NK cell of any one of the preceding claims, for use as a medicament.
The NK cell of any one of the preceding claims, for use in the treatment of a disease, disorder, or condition, e.g., a tumor and/or a cancer.
A method of increasing tumor killing ability of a NK cell, the method comprising:
- (a) knocking-into the genome of the NK cell a first exogenous coding sequence for FcγRIII (CD16) or variant thereof and a second exogenous coding sequence for a membrane bound interleukin 15 (mbIL-15), wherein the first exogenous coding sequence and the second exogenous coding sequence are knocked-in in frame and downstream (3′) of an essential gene;
- and
- (b) knocking-out one or more genes of the NK cell, wherein the one or more genes encode adenosine A2a receptor (ADORA2A), β-2 microglobulin (B2M), class II major histocompatibility complex transactivator (CIITA), cytokine inducible SH2 containing protein (CISH), two or more human leukocyte antigen (HLA) class II histocompatibility antigen alpha chain genes, two or more HLA class II histocompatibility antigen beta chain genes, natural killer group 2 member A receptor (NKG2A), programmed cell death protein 1 (PD-1), T cell immunoreceptor with Ig and ITIM domains (TIGIT), an agonist of the TGF beta signaling pathway (e.g., transforming growth factor beta receptor II (TGFβRII)), or any combination of two or more thereof;
- thereby increasing a level of tumor killing activity of the NK cell relative to a reference level of tumor killing activity of a reference NK cell.
A method of increasing antibody-dependent cellular cytotoxicity (ADCC) induced by a NK cell, the method comprising:
- (a) knocking-into the genome of the NK cell a first exogenous coding sequence for FcγRIII (CD16) or variant thereof and a second exogenous coding sequence for a membrane bound interleukin 15 (mbIL-15), wherein the first exogenous coding sequence and the second exogenous coding sequence are knocked-in in frame and downstream (3′) of an essential gene;
- and
- (b) knocking-out one or more genes of the NK cell, wherein the one or more genes encode adenosine A2a receptor (ADORA2A), β-2 microglobulin (B2M), class II major histocompatibility complex transactivator (CIITA), cytokine inducible SH2 containing protein (CISH), two or more human leukocyte antigen (HLA) class II histocompatibility antigen alpha chain genes, two or more HLA class II histocompatibility antigen beta chain genes, natural killer group 2 member A receptor (NKG2A), programmed cell death protein 1 (PD-1), T cell immunoreceptor with Ig and ITIM domains (TIGIT), an agonist of the TGF beta signaling pathway (e.g., transforming growth factor beta receptor II (TGFβRII)), or any combination of two or more thereof;
- thereby increasing a level of ADCC induced by the NK cell relative to a reference level of ADCC induced by a reference NK cell.
A method of increasing persistence of a NK cell, the method comprising:
- (a) knocking-into the genome of the NK cell a first exogenous coding sequence for FcγRIII (CD16) or variant thereof and a second exogenous coding sequence for a membrane bound interleukin 15 (mbIL-15), wherein the first exogenous coding sequence and the second exogenous coding sequence are knocked-in in frame and downstream (3′) of an essential gene;
- and
- (b) knocking-out one or more genes of the NK cell, wherein the one or more genes encode adenosine A2a receptor (ADORA2A), β-2 microglobulin (B2M), class II major histocompatibility complex transactivator (CIITA), cytokine inducible SH2 containing protein (CISH), two or more human leukocyte antigen (HLA) class II histocompatibility antigen alpha chain genes, two or more HLA class II histocompatibility antigen beta chain genes, natural killer group 2 member A receptor (NKG2A), programmed cell death protein 1 (PD-1), T cell immunoreceptor with Ig and ITIM domains (TIGIT), an agonist of the TGF beta signaling pathway (e.g., transforming growth factor beta receptor II (TGFβRII)), or any combination of two or more thereof;
- thereby increasing a level of persistence of the NK cell relative to a reference level of persistence of a reference NK cell.
A method of manufacturing a genetically modified NK cell, the method comprising:
- (a) knocking-into the genome of an NK cell a first exogenous coding sequence for FcγRIII (CD16) or variant thereof and a second exogenous coding sequence for a membrane bound interleukin 15 (mbIL-15), wherein the first exogenous coding sequence and the second exogenous coding sequence are knocked-in in frame and downstream (3′) of an essential gene;
- and
- (b) knocking-out one or more genes of the NK cell, wherein the one or more genes encode adenosine A2a receptor (ADORA2A), β-2 microglobulin (B2M), class II major histocompatibility complex transactivator (CIITA), cytokine inducible SH2 containing protein (CISH), two or more human leukocyte antigen (HLA) class II histocompatibility antigen alpha chain genes, two or more HLA class II histocompatibility antigen beta chain genes, natural killer group 2 member A receptor (NKG2A), programmed cell death protein 1 (PD-1), T cell immunoreceptor with Ig and ITIM domains (TIGIT), an agonist of the TGF beta signaling pathway (e.g., transforming growth factor beta receptor II (TGFβRII)), or any combination of two or more thereof.
Dependencies are inferred from the claim text.