Treatment of cancer using humanized anti-cd19 chimeric antigen receptor
AssigneesThe Trustees of the University of Pennsylvania, Novartis AG
InventorsJennifer Brogdon, Carl H. June, Andreas Loew, Marcela Maus, John Scholler
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Abstract
The invention provides compositions and methods for treating diseases associated with expression of CD19. The invention also relates to chimeric antigen receptor (CAR) specific to CD19, vectors encoding the same, and recombinant T cells comprising the CD19 CAR. The invention also includes methods of administering a genetically modified T cell expressing a CAR that comprises a CD19 binding domain.
Claims97
An isolated nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an antibody or antibody fragment which includes a humanized anti-CD19 binding domain, a transmembrane domain, and an intracellular signaling domain comprising a stimulatory domain, and wherein said anti-CD19 binding domain comprises one or more of light chain complementary determining region 1 (LC CDR1), light chain complementary determining region 2 (LC CDR2), and light chain complementary determining region 3 (LC CDR3) of any anti-CD19 light chain binding domain amino acid sequence listed in Table 3, and one or more of heavy chain complementary determining region 1 (HC CDR1), heavy chain complementary determining region 2 (HC CDR2), and heavy chain complementary determining region 3 (HC CDR3) of any anti-CD19 heavy chain binding domain amino acid sequence listed in Table 3.
The isolated nucleic acid molecule of any of the preceding claims, wherein the CD19 binding domain is a scFv.
The isolated nucleic acid molecule of any of the preceding claims, wherein the light chain variable region comprises an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications of an amino acid sequence of a light chain variable region provided in Table 3, or a sequence with 95-99% identity to an amino acid sequence provided in Table 3.
The isolated nucleic acid molecule of any of the preceding claims, wherein the heavy chain variable region comprises an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications of an amino acid sequence of a heavy chain variable region provided in Table 3, or a sequence with 95-99% identity to an amino acid sequence provided in Table 3.
The isolated nucleic acid molecule of any of the preceding claims, wherein the anti-CD19 binding domain comprises a sequence selected from a group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, and SEQ ID NO:12, or a sequence with 95-99% identify thereof.
The isolated nucleic acid molecule of any of the preceding claims, wherein the nucleic acid sequence encoding the anti-CD19 binding domain comprises a sequence selected from a group consisting of SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:70, SEQ ID NO:71 and SEQ ID NO:72, or a sequence with 95-99% identify thereof.
The isolated nucleic acid molecule of any of the preceding claims, wherein the encoded CAR includes a transmembrane domain that comprises a transmembrane domain of a protein selected from the group consisting of the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 and CD154.
The isolated nucleic acid molecule of any of the preceding claims, wherein the encoded transmembrane domain comprises a sequence of SEQ ID NO:
- 15.
The isolated nucleic acid molecule of any of the preceding claims, wherein the encoded transmembrane domain comprises an amino acid sequence comprises at least one, two or three modifications but not more than 20, 10 or 5 modifications of an amino acid sequence of SEQ ID NO:15, or a sequence with 95-99% identity to an amino acid sequence of SEQ ID NO:15.
The isolated nucleic acid molecule of any of the preceding claims, wherein the, the nucleic acid sequence encoding the transmembrane domain comprises a sequence of SEQ ID NO:56, or a sequence with 95-99% identify thereof.
The isolated nucleic acid molecule of any of the preceding claims, wherein the encoded anti-CD19 binding domain is connected to the transmembrane domain by a hinge region.
The isolated nucleic acid molecule of any of the preceding claims, further comprising a sequence encoding a costimulatory domain.
The isolated nucleic acid molecule of any of the preceding claims, further comprising a sequence encoding an intracellular signaling domain.
The isolated nucleic acid molecule of any of the preceding claims, wherein the encoded intracellular signaling domain comprises a functional signaling domain of 4-1BB and/or a functional signaling domain of CD3 zeta.
The isolated nucleic acid molecule of any of the preceding claims, wherein the encoded intracellular signaling domain comprises the sequence of SEQ ID NO:
- 16 and/or the sequence of SEQ ID NO:17 or SEQ ID NO:43.
The isolated nucleic acid molecule of any of the preceding claims, wherein the intracellular signaling domain comprises an amino acid sequence having at least one, two or three modifications but not more than 20, 10 or 5 modifications of an amino acid sequence of SEQ ID NO:16 and/or an amino acid sequence of SEQ ID NO:17 or SEQ ID NO:43, or a sequence with 95-99% identity to an amino acid sequence of SEQ ID NO:16 and/or an amino acid sequence of SEQ ID NO:17 or SEQ ID NO:43.
The isolated nucleic acid molecule of any of the preceding claims, wherein the encoded intracellular signaling domain comprises the sequence of SEQ ID NO:16 and the sequence of SEQ ID NO:17 or SEQ ID NO:43, wherein the sequences comprising the intracellular signaling domain are expressed in the same frame and as a single polypeptide chain.
The isolated nucleic acid molecule of any of the preceding claims, wherein the nucleic acid sequence encoding the intracellular signaling domain comprises a sequence of SEQ ID NO:60, or a sequence with 95-99% identify thereof, and/or a sequence of SEQ ID NO:101 or SEQ ID NO:44, or a sequence with 95-99% identity thereof.
The isolated nucleic acid molecule of any of the preceding claims, further comprising a leader sequence.
An isolated chimeric antigen receptor (CAR) molecule comprising a humanized anti-CD19 binding domain, a transmembrane domain, and an intracellular signaling domain.
A humanized anti-CD19 binding domain comprising one or more light chain complementary determining region 1 (LC CDR1), light chain complementary determining region 2 (LC CDR2), and light chain complementary determining region 3 (LC CDR3) of any anti-CD19 binding domain in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, and SEQ ID NO:12, and one or more heavy chain complementary determining region 1 (HC CDR1), heavy chain complementary determining region 2 (HC CDR2), and heavy chain complementary determining region 3 (HC CDR3) of any humanized anti-CD19 binding domain in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, and SEQ ID NO:12.
A vector comprising a nucleic acid molecule encoding a CAR of any of the preceding claims.
A method of generating a population of RNA-engineered cells comprising introducing an in vitro transcribed RNA or synthetic RNA into a cell, where the RNA comprises a nucleic acid encoding a CAR molecule of any of the preceding claims.
A method of providing an anti-tumor immunity in a mammal comprising administering to the mammal an effective amount of a cell expressing a CAR molecule of any of the preceding claims.
A method of treating a mammal having a disease associated with expression of CD19 comprising administering to the mammal an effective amount of the cell of comprising a CAR molecule of any of the preceding claims.
Dependencies are inferred from the claim text.