Chimeric antigen receptors specific for B cell maturation antigens for use in treating myeloma - Patent Application 20070123633

Abstract

Provided herein is a method of adoptive cell therapy involving administration of genetically engineered cells to treat diseases and conditions, including certain plasma cell malignancies. The cells generally express a recombinant receptor, such as a chimeric antigen receptor (CAR) specific for B cell maturation antigen (BCMA). In some embodiments, the method is for selecting and treating subjects with multiple myeloma (MM).

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority from U.S. Provisional Application No. 63 / 275,414, filed November 3, 2021, entitled "METHODS FOR TREATMENT USING CHIMERIC ANTIGEN RECEPTORS SPECIFIC FOR B-CELL MATURATION ANTIGEN," and U.S. Provisional Application No. 63 / 287,904, filed December 9, 2021, entitled "METHODS FOR TREATMENT USING CHIMERIC ANTIGEN RECEPTORS SPECIFIC FOR B-CELL MATURATION ANTIGEN," the contents of which are incorporated by reference in their entireties.

[0002] Incorporation by reference of sequence listing This application is submitted with an electronic Sequence Listing. The Sequence Listing is provided as a file entitled 683772001440SeqList.XML, created on November 2, 2022, and is 349,655 bytes in size. The information in the electronic format of the Sequence Listing is incorporated by reference in its entirety.

[0003] The present disclosure, in some aspects, relates to adoptive cell therapy for treating multiple myeloma (MM), which involves the administration of genetically engineered cells. The cells generally express a recombinant receptor, such as a chimeric antigen receptor (CAR) specific for B-cell maturation antigen (BCMA). In some embodiments, the present disclosure also relates to methods for selecting subjects with MM for treatment with CAR-expressing cells and predicting a subject's response to treatment therewith. [Background technology]

[0004] B-cell maturation antigen (BCMA) is a transmembrane type III protein expressed in mature B lymphocytes. After BCMA binds to its ligands, B cell of the TNF family (BAFF) or proliferation-inducing ligand (APRIL), a cell survival-promoting signal is delivered to B cells, which has been shown to be necessary for plasma cell survival. BCMA expression is associated with several diseases, including cancer, autoimmune disorders, and infectious diseases. The role of BCMA in various diseases and conditions, including cancer, has made it a therapeutic target. Various BCMA-binding chimeric antigen receptors (CARs) and cells expressing such CARs are available. However, there remains a need for improved methods of selecting subjects for use with BCMA-binding CARs and engineered BCMA-CAR-expressing target cells, such as in adoptive cell therapy, and for treating patients with them. Provided herein are embodiments that meet such needs. Summary of the Invention

[0005] Provided herein are methods of treating a subject with multiple myeloma (MM), comprising: (a) determining that the subject (i) has a serum soluble B-cell maturation antigen (sBCMA) level below about 600 ng / mL; and / or (ii) does not have the presence of IgG heavy chain disease (HCD); and (b) administering to the subject a T cell therapy containing a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds to human BCMA.

[0006] Also provided herein are methods of treating a subject having or suspected of having multiple myeloma (MM), comprising administering a T cell therapy containing a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds to human B-cell maturation antigen (BCMA) to the subject, who has been predetermined to have (i) a serum soluble B-cell maturation antigen (sBCMA) level below about 600 ng / mL, and / or (ii) no presence of IgG heavy chain disease (HCD).

[0007] Also provided herein are methods of treating a subject with multiple myeloma (MM), the method comprising: (a) selecting a subject for treatment with a T cell therapy containing a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds human BCMA, the subject being predetermined to (i) have a serum soluble B-cell maturation antigen (sBCMA) level below about 600 ng / mL and / or (ii) not have the presence of IgG heavy chain disease (HCD); and (b) administering the T cell therapy to the subject.

[0008] Also provided herein is a method of selecting a subject having multiple myeloma (MM) for treatment with a T cell therapy containing a dose of genetically engineered T cells, comprising determining that the subject (i) has a serum soluble B-cell maturation antigen (sBCMA) level below about 600 ng / mL; and / or (ii) does not have the presence of IgG heavy chain disease (HCD), wherein if the subject is determined to have (i) and / or (ii), the subject is selected for administration with the T cell therapy containing a dose of genetically engineered T cells expressing a chimeric antigen receptor (CAR) that binds to human BCMA.

[0009] Also provided herein is a method of predicting the response of a subject with multiple myeloma (MM) to treatment with a T cell therapy containing a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds to human B cell maturation antigen (BCMA), comprising determining that the subject (i) has a serum soluble B cell maturation antigen (sBCMA) level below about 600 ng / mL, and / or (ii) does not have the presence of IgG heavy chain disease (HCD), wherein if the subject has (i) and / or (ii), the subject is predicted to achieve a complete response (CR) or a stringent complete response (sCR), and wherein treatment comprises administering the dose of the engineered T cells to the subject.

[0010] Also provided herein are methods of treating a subject with multiple myeloma (MM), comprising: determining that the subject (i) has a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL and / or (ii) has IgG heavy chain disease (HCD); (b) debulking the MM; and (c) administering to the subject a T cell therapy containing a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds to human BCMA.

[0011] Also provided herein are methods of treating a subject with multiple myeloma (MM), comprising administering to the subject a T cell therapy containing a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds human B-cell maturation antigen (BCMA), wherein (a) the subject is previously determined to have (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL and / or (ii) the presence of IgG heavy chain disease (HCD), and (b) the MM has been debulked at a time between (1) the subject being determined to have (i) and / or (ii) and (2) the subject being administered the T cell therapy.

[0012] Also provided herein is a method of selecting a subject having multiple myeloma (MM) for debulking, comprising determining that the subject has (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL; and / or (ii) the presence of IgG heavy chain disease (HCD), wherein if the subject is determined to have (i) and / or (ii), the subject is selected for debulking of MM prior to administration to the subject of a T cell therapy containing a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds to human BCMA.

[0013] Also provided herein is a method of predicting the response of a subject with multiple myeloma (MM) to treatment with a T cell therapy containing a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds to human B cell maturation antigen (BCMA), comprising determining that the subject has (i) a serum soluble B cell maturation antigen (sBCMA) level greater than about 600 ng / mL, and / or (ii) the presence of IgG heavy chain disease (HCD), wherein if the subject has (i) and / or (ii), the subject is predicted to not achieve a complete response (CR) or a stringent complete response (sCR), and wherein treatment comprises administering the dose of the engineered T cells to the subject.

[0014] Provided herein are methods of treating a subject with multiple myeloma (MM), the methods including: (a) determining that the subject has a first serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL; (b) debulking the MM; (c) determining that the subject has a post-debulking serum sBCMA level less than about 600 ng / mL; and (d) administering to the subject a T cell therapy containing a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds to human BCMA.

[0015] In some embodiments, the subject is a human.

[0016] In some embodiments, the method further comprises determining that the subject (i) has a serum sBCMA level or a post-debulking serum sBCMA level greater than or less than about 600 ng / mL; and / or (ii) has or does not have the presence of IgG HCD. In some embodiments, the method comprises determining that the subject has a serum sBCMA level or a post-debulking serum sBCMA level greater than or less than about 600 ng / mL.

[0017] In some embodiments, the method includes determining that the subject has a serum soluble B-cell maturation antigen (sBCMA) level below about 600 ng / mL. In some embodiments, the method includes determining that the subject does not have the presence of IgG heavy chain disease (HCD). In some embodiments, the method includes determining that the subject has a serum soluble B-cell maturation antigen (sBCMA) level below about 600 ng / mL and does not have the presence of IgG heavy chain disease (HCD).

[0018] In some embodiments, the subject is determined to have a serum soluble B-cell maturation antigen (sBCMA) level below about 600 ng / mL. In some embodiments, the subject is determined to not have the presence of IgG heavy chain disease (HCD). In some embodiments, the subject is determined to have a serum soluble B-cell maturation antigen (sBCMA) level below about 600 ng / mL and not have the presence of IgG heavy chain disease (HCD).

[0019] In some embodiments, the method includes determining that the subject has a serum sBCMA level that is greater than or less than about 590ng / mL, about 580ng / mL, about 570ng / mL, about 560ng / mL, about 550ng / mL, about 540ng / mL, about 530ng / mL, about 520ng / mL, about 510ng / mL, or about 500ng / mL. In some embodiments, the subject is determined to have a serum sBCMA level that is greater than or less than about 590ng / mL, about 580ng / mL, about 570ng / mL, about 560ng / mL, about 550ng / mL, about 540ng / mL, about 530ng / mL, about 520ng / mL, about 510ng / mL, or about 500ng / mL.

[0020] In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 566 ng / mL. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 566 ng / mL. In some embodiments, determining that the subject has the presence of IgG HCD includes detecting IgG in the subject's serum. In some embodiments, determining that the subject has the presence of IgG HCD includes detecting IgG in the subject's urine.

[0021] In some embodiments, the determination of serum sBCMA levels is performed about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week before (i) administration of T cell therapy to the subject, or (ii) about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week before the dose of engineered CAR T cells is obtained from the subject. In some embodiments, the determination of serum sBCMA levels is performed about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week before the dose of engineered CAR T cells is obtained from the subject.

[0022] In some embodiments, the method includes determining whether the subject has or does not have the presence of IgG HCD. In some embodiments, determining whether the subject has the presence of IgG HCD includes detecting IgG in the subject's serum and / or urine.

[0023] In some embodiments, the determination that the subject has or does not have the presence of an IgG HCD is performed about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week before (i) administration of T cell therapy to the subject, or (ii) the dose of engineered CAR T cells is obtained from the subject. In some embodiments, the determination that the subject has or does not have the presence of an IgG HCD is performed about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week before administration of T cell therapy to the subject. In some embodiments, the determination that the subject has or does not have the presence of an IgG HCD is performed about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week before the dose of engineered CAR T cells is obtained from the subject.

[0024] In some embodiments, the methods include administering to a subject a dose of genetically engineered cells.

[0025] In some embodiments, if the subject is predicted to achieve CR or sCR, the method further comprises administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA.

[0026] In some embodiments, if the subject is predicted not to achieve CR or sCR, the method further comprises selecting the subject for debulking of MM prior to administering to the subject a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA.

[0027] In some embodiments, after administering a dose of the engineered T cells to the subject, the subject achieves a complete response (CR) or a stringent complete response (sCR). In some embodiments, after administering a dose of the engineered T cells to the subject, the subject achieves a CR. In some embodiments, after administering a dose of the engineered T cells to the subject, the subject achieves a sCR.

[0028] In some embodiments, debulking comprises administering chemotherapy, radiation, and / or an immunomodulatory agent to the subject. In some embodiments, debulking comprises administering chemotherapy to the subject. In some embodiments, debulking comprises administering radiation to the subject. In some embodiments, debulking comprises administering an immunomodulatory agent to the subject. In some embodiments, debulking comprises administering chemotherapy and radiation to the subject. In some embodiments, debulking comprises administering chemotherapy and an immunomodulatory agent to the subject. In some embodiments, debulking comprises administering radiation and an immunomodulatory agent to the subject. In some embodiments, debulking comprises administering chemotherapy, radiation, and an immunomodulatory agent to the subject.

[0029] In some embodiments, the chemotherapy is or includes melphalan, doxorubicin, or cyclophosphamide chemotherapy. In some embodiments, the chemotherapy is or includes melphalan. In some embodiments, the chemotherapy is or includes doxorubicin. In some embodiments, the chemotherapy is or includes cyclophosphamide chemotherapy. In some embodiments, the immunomodulatory agent is thalidomide, lenalidomide, or pomalidomide. In some embodiments, the immunomodulatory agent is thalidomide. In some embodiments, the immunomodulatory agent is lenalidomide. In some embodiments, the immunomodulatory agent is pomalidomide. In some embodiments, the immunomodulatory agent is a checkpoint inhibitor.

[0030] In some embodiments, debulking is performed within about 3 months, within about 2 months, within about 1 month, within about 3 weeks, within about 2 weeks, or within about 1 week prior to administration of the T cell therapy to the subject.

[0031] In some embodiments, prior to administration of a dose of engineered T cells to the subject, the subject administers fludarabine to a patient having a body surface area of ​​1 m 2 20-40 mg / m or approximately 20-40 mg / m, or 30 mg / m as appropriate 2 or approximately 30 mg / m2 daily for 2-4 days and / or cyclophosphamide per 1 m of body surface area of ​​the subject. 2 200-400 mg / m or approximately 200-400 mg / m, or 300 mg / m as appropriate 2 or approximately 300 mg / m 2 In some embodiments, prior to administration of the dose of engineered T cells to the subject, the subject is receiving lymphodepletion therapy comprising administering fludarabine to a patient having the disease at least once a day for 2-4 days. 2 30 mg per unit or approximately 30 mg / m 2 daily, cyclophosphamide for 1 m of body surface area of ​​the subject. 2 300 mg per unit or approximately 300 mg / m 2 He is undergoing lymphodepletion therapy, which involves administering 10 mg of ribozyme every day for three days.

[0032] In some embodiments, debulking is performed before lymphodepletion therapy. In some embodiments, debulking is performed after lymphodepletion therapy. In some embodiments, debulking is performed before lymphodepletion therapy and after lymphodepletion therapy. In some embodiments, the subject is treated with a gamma secretase inhibitor before administering a dose of genetically engineered T cells to the subject.

[0033] In some embodiments, the MM is high-risk MM, or relapsed and / or refractory (r / r) MM. In some embodiments, the MM is high-risk MM, or relapsed and refractory (r / r) MM. In some embodiments, the MM is high-risk MM. In some embodiments, the MM is relapsed and refractory (r / r) MM. In some embodiments, the MM is relapsed or refractory (r / r) MM. In some embodiments, the MM is high-risk MM and relapsed and refractory (r / r) MM.

[0034] In some embodiments, the subject is 18 years of age or older. In some embodiments, the subject has previously received three or more prior lines of therapy for MM. In some embodiments, each of the three or more prior lines of therapy included two consecutive cycles, unless progressive disease showed the best response to that line of therapy. In some embodiments, progressive disease is progression within 60 days after the last dose of a line of therapy. In some embodiments, the three or more prior lines of therapy include a proteasome inhibitor, an immunomodulatory agent, and an anti-CD38 antibody. In some embodiments, the subject is refractory to the last of the three or more prior lines of therapy. In some embodiments, the subject has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1. In some embodiments, the subject has an Eastern Cooperative Oncology Group (ECOG) performance status of 0. In some embodiments, the subject has an Eastern Cooperative Oncology Group (ECOG) performance status of 1.

[0035] In some embodiments, the subject has measurable disease at the time of administration of the dose of engineered T cells. In some embodiments, measurable disease includes (i) serum M protein ≧1.0 g / dL; (ii) urinary M protein ≧200 mg / 24 hours; and / or (iii) if serum free light chain (FLC) ratio is abnormal, diseased serum FLC level ≧10 mg / dL. In some embodiments, measurable disease includes serum M protein ≧1.0 g / dL. In some embodiments, measurable disease includes urinary M protein ≧200 mg / 24 hours. In some embodiments, measurable disease includes if serum free light chain (FLC) ratio is abnormal, diseased serum FLC level ≧10 mg / dL. In some embodiments, measurable disease includes (i) serum M protein ≧1.0 g / dL; and (ii) urinary M protein ≧200 mg / 24 hours. In some embodiments, measurable disease includes (i) serum M protein of 1.0 g or more; and (ii) if the serum free light chain (FLC) ratio is abnormal, a diseased serum FLC level of 10 mg / dL or more. In some embodiments, measurable disease includes (i) urinary M protein of 200 mg or more / 24 hours; and (ii) if the serum free light chain (FLC) ratio is abnormal, a diseased serum FLC level of 10 mg / dL or more. In some embodiments, measurable disease includes (i) serum M protein of 1.0 g or more; (ii) urinary M protein of 200 mg or more / 24 hours; and (iii) if the serum free light chain (FLC) ratio is abnormal, a diseased serum FLC level of 10 mg / dL or more.

[0036] In some embodiments, the subject has: (i) a central nervous system (CNS) lesion; and / or (ii) History or presence of clinically relevant CNS pathology In some embodiments, the subject does not have a central nervous system (CNS) lesion. In some embodiments, the subject does not have a history or presence of a clinically relevant CNS condition. In some embodiments, the subject does not have (i) a central nervous system (CNS) lesion; or (ii) History or presence of clinically relevant CNS pathology In some embodiments, the subject does not have or have a history of active plasma cell leukemia (PCL).

[0037] In some embodiments, the CAR contains an extracellular antigen-binding domain that binds to BCMA, a transmembrane domain, and an intracellular signaling region.

[0038] In some embodiments, the extracellular antigen-binding domain comprises a variable heavy chain (V H In some embodiments, the extracellular antigen-binding domain comprises or consists of a single domain antibody (sdAb). In some embodiments, the sdAb comprises a variable heavy chain (V H In some embodiments, the extracellular antigen-binding domain contains or consists of two sdAbs. In some embodiments, each of the two sdAbs is a variable heavy chain (V H ) region. In some embodiments, each of the two sdAbs binds to a different epitope of BCMA. In some embodiments, each of the two sdAbs binds to the same epitope of BCMA.

[0039] In some embodiments, the extracellular antigen-binding domain comprises a variable heavy chain (V H ) region and the variable light chain (V L ) region. In some embodiments, V H The regions contain CDR-H1, CDR-H2 and CDR-H3 containing the amino acid sequences set forth in SEQ ID NOs: 189, 190 and 191, respectively; and V L The region contains CDR-L1, CDR-L2 and CDR-L3 containing the amino acid sequences set forth in SEQ ID NOs: 192, 193 and 194, respectively: or V H The regions contain CDR-H1, CDR-H2 and CDR-H3 containing the amino acid sequences set forth in SEQ ID NOs: 173, 174 and 175, respectively; and V L The V region contains CDR-L1, CDR-L2 and CDR-L3, which contain the amino acid sequences set forth in SEQ ID NOs: 183, 184 and 185, respectively.H The regions contain CDR-H1, CDR-H2 and CDR-H3 containing the amino acid sequences set forth in SEQ ID NOs: 189, 190 and 191, respectively; and V L The V region contains CDR-L1, CDR-L2 and CDR-L3, which contain the amino acid sequences set forth in SEQ ID NOs: 192, 193 and 194, respectively. H The regions contain CDR-H1, CDR-H2 and CDR-H3 containing the amino acid sequences set forth in SEQ ID NOs: 173, 174 and 175, respectively; and V L The V region contains CDR-L1, CDR-L2 and CDR-L3, which contain the amino acid sequences set forth in SEQ ID NOs: 183, 184 and 185, respectively. H The region comprises the amino acid sequence set forth in SEQ ID NO: 18, L The region comprises the amino acid sequence set forth in SEQ ID NO: 19; or V H The region comprises the amino acid sequence set forth in SEQ ID NO: 24, L The region comprises the amino acid sequence set forth in SEQ ID NO: 25. H The region comprises the amino acid sequence set forth in SEQ ID NO: 18, L The region comprises the amino acid sequence set forth in SEQ ID NO: 19. H The region comprises the amino acid sequence set forth in SEQ ID NO: 24, L The region comprises the amino acid sequence set forth in SEQ ID NO:25.

[0040] In some embodiments, the extracellular antigen-binding domain is a single-chain variable fragment (scFv). In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO: 213 or SEQ ID NO: 188. In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO: 213. In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO: 188. In some embodiments, the scFv comprises the amino acid sequence set forth in any one of SEQ ID NOs: 216-247.

[0041] In some embodiments, the intracellular signaling region comprises the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain. In some embodiments, the intracellular signaling region further comprises a costimulatory signaling domain. In some embodiments, the costimulatory signaling domain comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof. In some embodiments, the costimulatory signaling domain comprises the intracellular signaling domain of CD28, or a signaling portion thereof. In some embodiments, the costimulatory signaling domain comprises the intracellular signaling domain of 4-1BB, or a signaling portion thereof. In some embodiments, the costimulatory signaling domain comprises the intracellular signaling domain of ICOS, or a signaling portion thereof.

[0042] In some embodiments, the costimulatory signaling domain is located between the transmembrane domain and the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain. In some embodiments, the transmembrane domain is or contains a transmembrane domain derived from CD28 or CD8. In some embodiments, the transmembrane domain is or contains a transmembrane domain derived from CD28. In some embodiments, the CD28 is human CD28. In some embodiments, the transmembrane domain is or contains a transmembrane domain derived from CD8. In some embodiments, the CD8 is human CD8.

[0043] In some embodiments, the CAR further comprises an extracellular spacer between the extracellular antigen-binding domain and the transmembrane domain. In some embodiments, the spacer is derived from CD8. In some embodiments, the spacer is a CD8α hinge.

[0044] In some embodiments, the CAR comprises the amino acid sequence set forth in any one of SEQ ID NOs: 90-141. In some embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 116 or SEQ ID NO: 124. In some embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 116. In some embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 124. In some embodiments, the CAR is encoded by the polypeptide sequence set forth in SEQ ID NO: 214.

[0045] In some embodiments, the dose of engineered T cells contains: idecabtagene autoleucel cells (e.g., ABECMA® cells); bb21217 cells; orvacabtagene autoleucel cells; CT103A cells; siltacabtagene autoleucel cells; KITE585 cells; CT053 cells; BCMA-CS1 cCAR (BC1cCAR) cells; P-BCMA-101 cells; P-BCMA-ALLO1 cells; C-CAR088 cells; Descartes-08 cells; PBCAR269A cells; ALLO-715 cells; PHE885 cells; AUTO8 cells; CTX120 cells; CB-011 cells; ALLO-605 (TuboCAR / MM) cells; pCDCAR1 (TriCAR-Z136) cells, or GC012F cells. In some embodiments, the dose of genetically engineered T cells contains idecbutagen-mediated ubiquitin-derived T cells (e.g., ABECMA® cells, etc.).

[0046] In some embodiments, the dose of engineered T cells is + In some embodiments, the dose of engineered T cells comprises CD4 + T cells or CD8 + In some embodiments, the dose of engineered T cells comprises a combination of CD4 + T cells and CD8 + In some embodiments, the dose of engineered T cells comprises a combination of CD4 + CAR-expressing T cells or CD8+ In some embodiments, the dose of engineered T cells comprises a combination of CD4 + CAR-expressing T cells and CD8 + In some embodiments, the combination of CAR-expressing T cells comprises CD4 + CAR-expressing T cells vs. CD8 + The ratio of CAR-expressing T cells is 1:1 or approximately 1:1. In some embodiments, CD4 + CAR-expressing T cells vs. CD8 + The ratio of CAR-expressing T cells is between 1:3 or approximately 1:3 and 3:1 or approximately 3:1. In some embodiments, CD4 + T cells vs. CD8 + The ratio of T cells is 1:1 or approximately 1:1. In some embodiments, CD4 + T cells vs. CD8 + The ratio of T cells is between 1:3 or approximately 1:3 and 3:1 or approximately 3:1.

[0047] In some embodiments, the percentage of naive-like T cells is greater than or about 60% of the total engineered T cells in the dose, optionally greater than or about 65%, 70%, 80%, 90%, or 95%. In some embodiments, the percentage of naive-like T cells is greater than or about 40% of the total CD4+ engineered T cells in the dose, optionally greater than or about 50%, 60%, 70%, 80%, 90%, or 95%. In some embodiments, the percentage of naive-like T cells is greater than or about 40% of the total CD8+ engineered T cells in the dose, optionally greater than or about 50%, 60%, 70%, 80%, 90%, or 95%. In some embodiments, the naive-like T cells are CCR7+CD45RA+, CD27+CCR7+, or CD62L-CCR7+. In some embodiments, the naive-like T cells are CCR7+CD45RA+. In some embodiments, the naive-like T cells are CD27+CCR7+. In some embodiments, the naive-like T cells are CD62L-CCR7+.

[0048] In some embodiments, the percentage of central memory T cells is greater than or about 60% of the total engineered T cells in the dose, optionally greater than or about 65%, 70%, 80%, 90%, or 95%. In some embodiments, the percentage of central memory T cells is greater than or about 40% of the total CD4+ engineered T cells in the dose, optionally greater than or about 50%, 60%, 70%, 80%, 90%, or 95%. In some embodiments, the percentage of central memory T cells is greater than or about 40% of the total CD8+ engineered T cells in the dose, optionally greater than or about 50%, 60%, 70%, 80%, 90%, or 95%.

[0049] In some embodiments, the dose of engineered T cells is about 0.5 x 10 6 ~About 600×10 6 In some embodiments, the dose of engineered T cells contains about 0.5 x 10 CAR-positive T cells. 6 ~About 100×10 6 In some embodiments, the dose of engineered T cells contains about 0.5 x 10 CAR-positive T cells. 6 ~About 10×10 6 In some embodiments, the dose of engineered T cells contains about 1 x 10 CAR-positive T cells. 6 ~About 10×10 6 In some embodiments, the dose of engineered T cells contains about 0.5 x 10 CAR-positive T cells. 6 ~Approx. 1×10 6 In some embodiments, the dose of engineered T cells contains about 0.5 x 10 CAR-positive T cells. 6 In some embodiments, the dose of engineered T cells contains about 0.75 x 10 CAR-positive T cells. 6 In some embodiments, the dose of engineered T cells contains about 1.0 x 10 CAR-positive T cells. 6In some embodiments, the dose of engineered T cells contains about 2.5 x 10 CAR-positive T cells. 6 In some embodiments, the dose of engineered T cells contains about 5.0 x 10 CAR-positive T cells. 6 In some embodiments, the dose of engineered T cells contains about 7.5 x 10 CAR-positive T cells. 6 In some embodiments, the dose of engineered T cells contains about 10.0 x 10 CAR-positive T cells. 6 In some embodiments, the dose of engineered T cells contains about 50 x 10 CAR-positive T cells. 6 ~About 1000×10 6 In some embodiments, the dose of engineered T cells contains about 100 x 10 CAR-positive T cells. 6 ~About 600×10 6 In some embodiments, the dose of engineered T cells contains about 150 x 10 CAR-positive T cells. 6 ~Approx. 450×10 6 In some embodiments, the dose of engineered T cells contains about 150 x 10 CAR-positive T cells. 6 , 300×10 6 , or approximately 450 × 10 6 In some embodiments, the dose of engineered T cells contains about 150 x 10 CAR-positive T cells. 6 In some embodiments, the dose of engineered T cells contains about 300 x 10 CAR-positive T cells. 6 In some embodiments, the dose of engineered T cells contains about 450 x 10 CAR-positive T cells. 6 Contains CAR-positive T cells.

[0050] In some embodiments, the dose of engineered CAR T cells is obtained from a subject. In some embodiments, the dose of engineered CAR T cells is obtained from a subject, e.g., for genetic engineering. In some embodiments, the dose of engineered T cells is autologous to the subject. In some embodiments, the dose of engineered T cells is allogeneic to the subject. [Brief explanation of the drawings]

[0051] [Figure 1] Correlations between baseline characteristics and increased likelihood of CR / sCR (right) or non-CR / sCR (left) are shown ranked by importance (each dot represents an individual patient; dark gray: high or present levels; light gray: low or absent levels; CR: complete response; sCR: stringent complete response). a From patient laboratory measurements. [Figure 2A] Figure 2A shows the association between baseline soluble BCMA (sBCMA) levels and clinical response (CR: complete response; sCR: stringent complete response; VGPR: very good partial response; PR: partial response). Boxes represent Q1, Q3, and median values. Whiskers represent minimum values ​​greater than Q1 - 1.5*IQR and maximum values ​​less than Q3 + 1.5*IQR. [Figure 2B] Figure 2B shows the association between baseline levels of beta-2 microglobulin and clinical response (CR: complete response; sCR: stringent complete response; VGPR: very good partial response; PR: partial response). Boxes represent Q1, Q3, and the median. Whiskers represent the minimum value greater than Q1 - 1.5*IQR and the maximum value less than Q3 + 1.5*IQR. [Figure 2C] Figure 2C shows the number of patients with and without IgG heavy chain disease who showed CR / sCR or non-CR / sCR (CR: complete response; sCR: stringent complete response). [Figure 2D]Figure 2D shows the association between baseline levels of D-dimer, ferritin, and sodium and clinical response (CR: complete response; sCR: stringent complete response; VGPR: very good partial response; PR: partial response). Boxes represent Q1, Q3, and median values. Whiskers represent minimum values ​​greater than Q1 - 1.5*IQR and maximum values ​​less than Q3 + 1.5*IQR. DETAILED DESCRIPTION OF THE INVENTION

[0052] Provided herein are methods of treating a subject with multiple myeloma (MM) with a T cell therapy, e.g., a T cell therapy that targets BCMA, such as comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA, wherein the subject has a serum soluble BCMA (sBCMA) level below about 600 ng / ml and / or is determined to not have the presence of IgG heavy chain disease (HCD). In some embodiments, the subject is selected for administration of the T cell therapy.

[0053] Also provided herein are methods of treating a subject with multiple myeloma (MM) with T cell therapy, e.g., T cell therapy that targets BCMA, such as comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein the subject has a serum soluble BCMA (sBCMA) level greater than about 600 ng / ml and / or is determined to have the presence of IgG heavy chain disease (HCD). In some embodiments, the subject is selected for debulking of the MM and administration of T cell therapy. In some embodiments, the subject is selected for debulking of the MM. In some embodiments, the MM is debulked prior to administration of the T cell therapy.

[0054] In some embodiments, methods of selecting patients for administration of T cell therapy are based on determining that the subject has a serum level of sBCMA below a particular threshold, such as below about 600 ng / mL, and / or does not have the presence of IgG heavy chain disease. In some embodiments, methods of selecting patients for debulking of MM are based on determining that the subject has a serum level of sBCMA above a particular threshold, such as above about 600 ng / mL, and / or has the presence of IgG heavy chain disease.

[0055] Also provided herein are methods for predicting the likelihood that a subject will respond to treatment with a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds to BCMA. In some embodiments, the response is a complete response (CR) or a stringent complete response (sCR). In some embodiments, the likelihood that a subject will exhibit a CR or sCR to treatment with a T cell therapy is based on determining that the subject has a serum level of soluble BCMA below a particular threshold, such as below about 600 ng / mL, and / or does not have the presence of IgG heavy chain disease. In some embodiments, the subject is predicted to achieve a CR or sCR in response to treatment with a T cell therapy based on determining that the subject has a serum level of soluble BCMA below a particular threshold, such as below about 600 ng / mL, and / or does not have the presence of IgG heavy chain disease. In some embodiments, based on determining that a subject has a serum level of soluble BCMA above a particular threshold, for example, above about 600 ng / mL, and / or has the presence of IgG heavy chain disease, the subject is predicted not to achieve a CR or sCR in response to treatment with the T cell therapy.

[0056] Among the embodiments provided, methods, compositions, articles of manufacture, methods and uses, including those that target or direct BCMA and BCMA-expressing cells (i.e., multiple myeloma), are particularly mentioned. BCMA is expressed in malignant plasma cells, such as from any relapsed or newly diagnosed myeloma patient, with little or no expression observed in normal tissues. Among the embodiments provided, approaches useful for treating subjects with multiple myeloma and the selection of subjects with multiple myeloma for treatment with T cell therapy, including genetically engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, as well as compositions and articles of manufacture comprising the same, are particularly mentioned. BCMA CAR T cells generally express antibodies specific for BCMA (heavy chain variable (V) H In some embodiments, the antigen-binding antibody fragments, including scFvs, comprise an extracellular antigen-binding domain, such as a V 1 domain, a V 2 domain, a V 3 domain, a V 4 domain, a V 5 domain, a V 6 domain, a V 7 domain, a V 8 domain, a V 9 domain, a V 10 domain, a V 11 domain, a V 12 domain, a V 13 domain, a V 14 domain, a V 15 domain, a V H and light chain variable (V L ) region. Cells, such as engineered or recombinant cells, that express such anti-BCMA CARs and / or contain nucleic acids encoding such anti-BCMA CARs, as well as compositions and articles of manufacture and therapeutic doses containing such cells, are also provided.

[0057] Also provided are methods of selecting subjects with multiple myeloma for debulking of MM prior to administration of T cell therapy comprising BCMA CAR T cells. In some cases, the subject is selected for debulking if the subject has a serum soluble BCMA (sBCMA) level above a certain threshold, for example, above about 600 ng / mL, and / or is determined to have the presence of IgG heavy chain disease (HCD).

[0058] Also provided are methods of predicting whether a subject with multiple myeloma will achieve a complete response (CR) or a stringent complete response (sCR) after administration of a T cell therapy comprising BCMA CAR T cells to the subject. In some cases, the predictive method is based on serum soluble BCMA (sBCMA) levels, such as in a sample obtained from the patient prior to administration of the T cell therapy, and / or the presence or absence of IgG heavy chain disease (HCD).

[0059] Adoptive cell therapy (including the administration of BCMA CAR T cells for the treatment of multiple myeloma, as well as other adoptive immune cell and T cell therapies) can be effective in the treatment of cancer and other diseases and disorders. However, in certain situations, the available approaches to adoptive cell therapy may not always be entirely satisfactory.

[0060] In some embodiments, available approaches to treating multiple myeloma (e.g., relapsed and refractory MM) are complex and may not always be completely satisfactory. Patients with relapsed or refractory MM have poor outcomes with currently available therapies. Relapsed and refractory MM often do not respond to further treatment and typically progress within 2 to 4 months. (Chari et al., N Engl J Med (2019) 381:727-38 and Lonial et al., Lancet Oncol (2020) 21:207-21). In some embodiments, the choice of treatment regimen may depend on numerous factors, including drug availability, response to prior therapy, aggressiveness of relapse, eligibility for autologous stem cell transplantation (ASCT), and whether relapse occurred during or outside of treatment. In some embodiments, because MM relapses and remissions, existing regimens may, in some cases, result in relapse and / or treatment-related toxicity. In some cases, subjects with particularly aggressive disease, such as those whose disease has persisted or recurred after various treatments, those with a high disease burden, such as a high tumor burden, and / or those with high-risk disease (i.e., high-risk cytogenetics), may be particularly difficult to treat, and responses to certain treatments in these subjects may be poor or short-lasting. In some cases, heavily pretreated subjects, for example, those who have relapsed after several different lines of prior therapy, may exhibit low response rates and / or high incidence of adverse events.

[0061] In particular, patients with relapsed and / or refractory multiple myeloma (R / R MM) who have previously been exposed to immunomodulators, proteasome inhibitors (PIs), and anti-CD38 antibodies have poor outcomes (Chari et al., N Engl J Med (2019) 381:727-38; Lonial et al., Poster presentation at the European Hematology Association (EHA) Virtual Meeting 2021: Abstract EP970; and Richardson et al., J Clin Oncol (2021) 39:757-67). Furthermore, it is currently difficult to predict which patients will achieve a deep response to CAR T-cell treatment. In some aspects, provided embodiments are based on the observation that selection of subjects for treatment or therapy according to provided embodiments results in high response rates (i.e., CR or sCR), including subjects who have been exposed to prior lines of therapy (e.g., immunomodulators, proteasome inhibitors (PIs), and anti-CD38 antibodies). In particular, provided embodiments are based on the observation that subjects are more likely to or predicted to achieve CR or sCR in response to treatment with BCMA CAR T cells if the subjects have serum sBCMA levels below a certain threshold (e.g., below 500, 566, or 600 ng / mL) and / or if the subjects do not have the presence of IgG heavy chain disease (HCD). Conversely, it is observed herein that if a subject has serum sBCMA levels above a certain threshold (e.g., above 500, 566, or 600 ng / mL), and / or if the subject has the presence of IgG heavy chain disease (HCD), then the subject is predicted to be less likely or not to achieve a CR or sCR in response to treatment with BCMA CAR T cells.Based on these observations, it is contemplated that subjects with serum sBCMA levels above a certain threshold (e.g., above 500, 566, or 600 ng / mL) and / or with the presence of IgG heavy chain disease (HCD) may be selected for and / or subjected to debulking of MM prior to administration of T cell therapy. In some cases, debulking of MM prior to administration of T cell therapy results in the subject's serum sBCMA level being below a certain threshold (e.g., below 500, 566, or 600 ng / mL). In some cases, after debulking, the subject is administered T cell therapy.

[0062] The provided embodiments are based on observations from clinical studies that, in some circumstances, administration of BCMA CAR T cells such as those described herein results in higher CR or sCR rates in subjects with lower serum sBCMA levels and / or without the presence of IgG HCD, including relapsed and refractory subjects who have received three or more prior lines of therapy. In some aspects, the provided methods, uses, and cells result in high CR or sCR rates. In some aspects, such high responses can be achieved from selecting subjects with serum sBCMA levels below a certain threshold (e.g., below 500, 566, or 600 ng / mL) and / or determined to not have the presence of IgG heavy chain disease (HCD) for administration of T cell therapy. Furthermore, such high responses can be achieved from selecting subjects with serum sBCMA levels above a certain threshold (e.g., above 500, 566, or 600 ng / mL) and / or determined to have the presence of IgG heavy chain disease (HCD) for debulking of MM prior to administration of T cell therapy. In some aspects, treatment of subjects with high-risk and / or relapsed and refractory MM (e.g., heavily pretreated subjects, subjects with high tumor burden, and / or subjects with high cytogenetic risk) according to provided embodiments has been observed to provide effective and durable treatment, resulting in CR or sCR.

[0063] In various embodiments, the provided methods enable the selection and / or treatment of subjects with high-risk and / or R / R MM, overcoming or counteracting certain limitations that may reduce optimal response to cell therapy in such subjects. In some situations, the use of the provided methods and engineered cells or compositions comprising engineered cells has been observed to provide advantages in treating subjects, resulting in high CR or sCR rates at a variety of different dose levels tested. Furthermore, the use of the provided methods and engineered cells or compositions comprising engineered cells has been observed to provide advantages in treating subjects with particularly high-risk and / or R / R disease, including subjects who have relapsed and are refractory to multiple different prior treatments for the disease.

[0064] All publications, including patent documents, scientific articles, and databases, referenced in this application are incorporated herein by reference in their entirety for all purposes to the same extent as if each individual publication was individually incorporated herein by reference. To the extent that a definition set forth herein contradicts or is otherwise inconsistent with a definition set forth in a patent, application, published application, or other publication incorporated herein by reference, the definition set forth herein takes precedence over the definition incorporated herein by reference.

[0065] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described.

[0066] I. Methods of assessing biomarkers to select subjects for and predict response to T cell therapy treatment Among the methods and uses provided are methods for selecting a subject for treatment with T cell therapy, which may include assessing or detecting, e.g., in a sample from the subject, a biomarker or parameter associated with a response outcome to T cell therapy treatment (e.g., complete response (CR) or stringent complete response (sCR)). Also provided are methods for selecting a subject for debulking treatment and methods for predicting a subject's response to T cell therapy treatment, which may include assessing or detecting, e.g., in a sample from the subject, a biomarker or parameter associated with a response outcome to T cell therapy treatment (e.g., complete response (CR) or stringent complete response (sCR)). Among the methods and uses provided are methods for predicting whether a subject will exhibit a response outcome to T cell therapy treatment, which may include assessing or detecting, e.g., in a sample from the subject, a biomarker or parameter associated with a response outcome to T cell therapy treatment (e.g., complete response (CR) or stringent complete response (sCR)).

[0067] Complete response (CR) and stringent complete response (sCR) were defined according to the International Myeloma Working Group (IMWG) standard response criteria (Kumar et al., Lancet Oncol (2016) 17(8):e328-46) as shown in Table 1 below.

[0068] [Table 1] **All recommendations for clinical use regarding serum FLC levels or FLC ratios are based on results obtained in the validated Freelite test (Binding Site, Birmingham, UK). ††The presence / absence of clonal cells in immunohistochemistry is based on the kappa / lambda / L ratio. An abnormal kappa / lambda ratio by immunohistochemistry requires a minimum of 100 plasma cells for analysis. An abnormal ratio reflects the presence of an abnormal clone: ​​kappa / lambda > 4:1 or < 1:2.

[0069] In some of the embodiments, the method includes assessing the level of serum soluble B-cell maturation antigen (sBCMA), such as in a sample obtained from the subject. In some embodiments, the sample is a serum sample. In some of the embodiments, the method includes determining whether the subject has a serum sBCMA level below about 600 ng / mL. In some of the embodiments, the method includes determining whether the subject has a serum sBCMA level above about 600 ng / mL. In some of the embodiments, the method includes determining whether the subject has a serum sBCMA level below about 566 ng / mL. In some of the embodiments, the method includes determining whether the subject has a serum sBCMA level above about 566 ng / mL. In some of the embodiments, the method includes determining whether the subject has a serum sBCMA level below about 500 ng / mL. In some of the embodiments, the method includes determining whether the subject has a serum sBCMA level above about 500 ng / mL. In some embodiments, the method includes individually comparing the level, amount, or concentration of serum sBCMA from a sample obtained from the subject to a threshold level, thereby determining the likelihood that the subject will achieve a CR or sCR to the T cell therapy. In some aspects, the threshold level is determined based on the mean or median serum sBCMA level, amount, or concentration, and values ​​within a range or standard deviation of the mean or median, in biological samples obtained from a group of subjects prior to receiving T cell therapy, wherein each of the subjects in the group either went on to exhibit a CR or sCR or did not exhibit a CR or sCR.

[0070] In some of any of the embodiments, the method includes assessing whether a patient has the presence of IgG heavy chain disease (HCD), such as in a sample obtained from the subject. In some embodiments, the sample is a serum sample or a urine sample. In some of any of the embodiments, the method includes determining whether the subject has the presence of IgG HCD. In some of any of the embodiments, the method includes determining whether the subject does not have the presence of IgG HCD.

[0071] In some of the embodiments, the method includes assessing the copy number of a vector in a dose of engineered cells. In some embodiments, the vector comprises or encodes a chimeric antigen receptor (CAR). In some embodiments, the method includes comparing the copy number of the vector in a dose of engineered cells with a threshold level, thereby determining the likelihood that the subject will achieve CR or sCR to T cell therapy.

[0072] In some of any of the embodiments, the method includes assessing beta-2 microglobulin levels, such as in a sample obtained from the subject. In some embodiments, the sample is a blood sample. In some embodiments, the sample is a urine sample. In some embodiments, the sample is a cerebrospinal fluid (CSF) sample. In some embodiments, the method includes individually comparing the level, amount, or concentration of beta-2 microglobulin from the sample obtained from the subject to a threshold level, thereby determining the likelihood that the subject will achieve CR or sCR to the T cell therapy.

[0073] In some of any of the embodiments, the method includes assessing D-dimer levels, such as in a sample obtained from the subject. In some embodiments, the sample is a blood sample. In some embodiments, the method includes comparing the level, amount, or concentration of D-dimer from the sample obtained from the subject individually to a threshold level, thereby determining the likelihood that the subject will achieve CR or sCR to the T cell therapy.

[0074] In some of any of the embodiments, the method includes assessing ferritin levels, such as in a sample obtained from the subject. In some embodiments, the sample is a blood sample. In some embodiments, the method includes comparing the level, amount, or concentration of ferritin from the sample obtained from the subject individually to a threshold level, thereby determining the likelihood that the subject will achieve CR or sCR to the T cell therapy.

[0075] In some of any of the embodiments, the method includes assessing sodium levels, such as in a sample obtained from the subject. In some embodiments, the sample is a blood sample. In some embodiments, the method includes individually comparing the level, amount, or concentration of sodium from the sample obtained from the subject to a threshold level, thereby determining the likelihood that the subject will achieve CR or sCR to the T cell therapy.

[0076] In some of any of the embodiments, the method includes assessing the prothrombin time-international normalized ratio (PT-INR), such as in a sample obtained from the subject. In some embodiments, the sample is a blood sample. In some embodiments, the method includes comparing the PT-INR from the sample obtained from the subject to a threshold PT-INR, thereby determining the likelihood that the subject will achieve a CR or sCR to the T cell therapy.

[0077] In some embodiments, the T cell therapy comprises a dose of engineered cells expressing a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells). In some embodiments, serum sBCMA levels and / or the presence or absence of IgG HCD are assessed in a sample obtained from or from a subject with multiple myeloma, such as relapsed and refractory (RR) multiple myeloma. In some embodiments, the subject is a candidate for and / or has been treated with T cell therapy. In some embodiments, the provided methods can be used to identify or select subjects likely to respond to T cell therapy; and / or to select subjects for a particular treatment prior to administration of T cell therapy, such as debulking.

[0078] In some aspects, the methods include further monitoring the subject for a likely response based on the likelihood of response determined according to the provided embodiments, e.g., by assessment of serum sBCMA and / or IgG HCD.

[0079] In some embodiments, the methods include assessing or detecting the presence or absence of IgG HCD and / or the concentration, amount, or level of serum sBCMA. In some cases, the methods may include comparing the concentration, amount, or level of serum sBCMA to a particular reference value, such as a threshold level, e.g., a reference value associated with a particular response, such as CR and / or sCR. In some embodiments, the methods also include selecting subjects for treatment with T cell therapy based on assessing the presence or absence of IgG HCD and / or comparing serum sBCMA to a reference or threshold level of serum sBCMA.

[0080] In some embodiments, a biological sample, such as a serum or urine sample from a subject, can be obtained to detect the presence or absence of IgG HCD. In some embodiments, a biological sample, such as a blood or serum sample from a subject, can be obtained to detect the concentration, amount, or level of serum sBCMA.

[0081] In some embodiments, IgG HCD and / or serum sBCMA levels are objectively measurable characteristics or molecules expressed by or in a biological sample containing cells that can indicate or correlate with a particular condition or phenomenon, such as a treatment outcome or a disease state. In some aspects, IgG HCD and / or serum sBCMA can be measured or detected. For example, the presence or absence of IgG HCD can be detected. In some aspects, a parameter such as serum sBCMA concentration, amount, or level can be measured or detected. In some embodiments, the presence or absence of IgG HCD and / or serum sBCMA concentration, amount, or level can be associated with, correlated with, indicative of, and / or predict a particular condition, such as a particular treatment outcome or condition, in a subject. In some aspects, the presence or absence of IgG HCD and / or serum sBCMA concentration, amount, or level can be used to assess the likelihood of a particular outcome or condition, such as a particular treatment outcome, including a response outcome. In some embodiments, the response outcome is a complete response (CR) or a stringent complete response (sCR). In some embodiments, the response outcome is a CR. In some embodiments, the response outcome is sCR. Thus, in some embodiments, the presence or absence of IgG HCD and / or the concentration, amount, or level of serum sBCMA can be used to assess the likelihood that a subject will exhibit CR or sCR following administration of a T cell therapy.

[0082] In some embodiments, the presence or absence of IgG HCD can be used alone or in combination with the concentration, amount, or level of serum sBCMA. In some embodiments, the concentration, amount, or level of serum sBCMA can be used alone or in combination with the presence or absence of IgG HCD.

[0083] In some embodiments, the presence or absence of IgG HCD and / or serum sBCMA levels are determined from a biological sample. In some aspects, the biological sample is a body fluid or tissue. In some such embodiments, the biological sample, e.g., a body fluid, is or contains whole blood, serum, or plasma. In some such embodiments, the biological sample, e.g., a body fluid, is or contains serum. In some such embodiments, the biological sample, e.g., a body fluid, is or contains urine.

[0084] In some embodiments, the presence or absence of IgG HCD and / or serum sBCMA levels are determined prior to administration of the T cell therapy (e.g., pre-infusion), for example, obtained up to 1 day, up to 2 days, up to 7 days, up to 14 days, up to 21 days, up to 28 days, up to 35 days, up to 40 days, up to 2 months, or up to 3 months prior to initiation of administration of the T cell therapy.

[0085] In some embodiments, the presence or absence of an IgG HCD is determined prior to administration of the T cell therapy (e.g., prior to infusion), e.g., up to 1 day, 2 days, 7 days, 14 days, 21 days, 28 days, 35 days, 40 days, 2 months, or 3 months prior to the start of administration of the T cell therapy. In some embodiments, the determination of whether a subject has or does not have the presence of an IgG HCD is performed about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week prior to administration of the T cell therapy to the subject. In some embodiments, the determination of whether a subject has or does not have the presence of an IgG HCD is performed about 3 months prior to administration of the T cell therapy to the subject. In some embodiments, the determination of whether a subject has or does not have the presence of an IgG HCD is performed about 2 months prior to administration of the T cell therapy to the subject. In some embodiments, the determination of whether a subject has or does not have the presence of an IgG HCD is performed about three weeks prior to administration of T cell therapy to the subject. In some embodiments, the determination of whether a subject has or does not have the presence of an IgG HCD is performed about two weeks prior to administration of T cell therapy to the subject. In some embodiments, the determination of whether a subject has or does not have the presence of an IgG HCD is performed about one week prior to administration of T cell therapy to the subject. In some embodiments, the determination of whether a subject has or does not have the presence of an IgG HCD is performed before leukapheresis. In some embodiments, the determination of whether a subject has or does not have the presence of an IgG HCD is performed before administration of lymphodepletion therapy to the subject.

[0086] In some embodiments, the level of serum sBCMA is determined prior to administration of the T cell therapy (e.g., pre-infusion), e.g., obtained up to 1 day, 2 days, 7 days, 14 days, 21 days, 28 days, 35 days, 40 days, 2 months, or 3 months prior to initiation of administration of the T cell therapy. In some embodiments, a first level of serum sBCMA is determined prior to administration of the T cell therapy (e.g., pre-infusion), e.g., obtained up to 1 day, 2 days, 7 days, 14 days, 21 days, 28 days, 35 days, 40 days, 2 months, or 3 months prior to initiation of administration of the T cell therapy. In some embodiments, the debulking level of serum sBCMA is determined prior to administration of the T cell therapy (e.g., pre-infusion), for example, obtained up to 1 day, up to 2 days, up to 7 days, up to 14 days, up to 21 days, up to 28 days, up to 35 days, up to 40 days, up to 2 months, or up to 3 months prior to initiation of administration of the T cell therapy.

[0087] In some embodiments, the determination of serum sBCMA level is performed about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week prior to administration of T cell therapy to the subject. In some embodiments, the determination of serum sBCMA level is performed about 3 months prior to administration of T cell therapy to the subject. In some embodiments, the determination of serum sBCMA level is performed about 2 months prior to administration of T cell therapy to the subject. In some embodiments, the determination of serum sBCMA level is performed about 1 month prior to administration of T cell therapy to the subject. In some embodiments, the determination of serum sBCMA level is performed about 3 weeks prior to administration of T cell therapy to the subject. In some embodiments, the determination of serum sBCMA level is performed about 2 weeks prior to administration of T cell therapy to the subject. In some embodiments, the determination of serum sBCMA level is performed about 1 week prior to administration of T cell therapy to the subject. In some embodiments, the determination of serum sBCMA level is performed prior to leukapheresis. In some embodiments, the determination of serum sBCMA level is performed prior to administration of lymphodepleting therapy to the subject.

[0088] In some embodiments, the biological sample is obtained from the subject prior to administration of the cell therapy (e.g., prior to infusion), for example, up to 1 day, 2 days, 7 days, 14 days, 21 days, 28 days, 35 days, 40 days, 2 months, or 3 months prior to initiation of administration of the T cell therapy. In some embodiments, the biological sample is obtained from the subject prior to leukapheresis. In some embodiments, the biological sample is obtained from the subject prior to administration of lymphodepletion therapy to the subject.

[0089] In some embodiments, the serum sBCMA level is determined in a biological sample. In some embodiments, the biological sample is a blood, serum, or plasma sample. In some embodiments, the biological sample is a blood sample. In some embodiments, the biological sample is an apheresis or leukapheresis sample. In some embodiments, the biological sample is a serum sample. In some embodiments, the serum sBCMA level is determined in a serum sample from a subject.

[0090] In some embodiments, the presence or absence of IgG HCD is determined in a biological sample. In some embodiments, the biological sample is a blood sample, a serum sample, a plasma sample, or a urine sample. In some embodiments, the biological sample is a blood sample. In some embodiments, the biological sample is an apheresis or leukapheresis sample. In some embodiments, the biological sample is a serum sample. In some embodiments, the presence or absence of IgG HCD is determined in a serum sample from a subject. In some embodiments, the biological sample is a urine sample. In some embodiments, the presence or absence of IgG HCD is determined in a urine sample from a subject.

[0091] In some embodiments, the reagents can be used prior to administration of T cell therapy or after administration of T cell therapy for diagnostic purposes, to identify subjects and / or to assess the outcome of treatment.

[0092] In some embodiments, determining the presence or absence of IgG HCD and / or the level of serum sBCMA comprises performing an in vitro assay. In some embodiments, determining the presence or absence of IgG HCD comprises performing an in vitro assay. In some embodiments, determining the level of serum sBCMA comprises performing an in vitro assay. In some aspects, the in vitro assay is an immunoassay, an aptamer-based assay, a histological or cytological assay, or an mRNA expression level assay. In some embodiments, determining comprises measuring by enzyme-linked immunosorbent assay (ELISA), immunoblotting, immunoprecipitation, radioimmunoassay (RIA), immunostaining, flow cytometry assay, surface plasmon resonance (SPR), chemiluminescence assay, lateral flow immunoassay, inhibition assay, or avidity assay. In some cases, determining comprises using a binding reagent that specifically binds to IgG. In some aspects, determining comprises using a binding reagent that specifically binds to BCMA, e.g., sBCMA. In some aspects, the binding reagent is an antibody or antigen-binding fragment thereof, an aptamer, or a nucleic acid probe.

[0093] In some embodiments, tumor-associated BCMA expression is assessed from a bone marrow biopsy. In some embodiments, tumor-associated BCMA expression is assessed immunohistochemically using an antibody, such as a monoclonal antibody, directed against an intracellular BCMA epitope. In some embodiments, tumor cell surface BCMA expression is assessed using Quantibrite (商標)Soluble BCMA is quantified in fresh bone marrow aspirates by flow cytometry, such as using beads. In some embodiments, soluble BCMA is assessed in serum, such as using a Luminex® immunoassay. In some embodiments, soluble BCMA is assessed longitudinally (e.g., on consecutive days or at regular intervals) in serum. Methods for measuring soluble BCMA, including serum BCMA, are known in the art and can include any of the methods described in Munshi et al., N Engl J Med (2021) 384:705-16. sBCMA levels may be determined by an ONCOtracker assay (oncotracker.com / sbcma-biomarker / ).

[0094] In some embodiments, IgG heavy chain disease is determined by any method known in the art, including serum or urine immunofixation.

[0095] A. Serum soluble BCMA i. Choices regarding treatment and / or debulking In some embodiments, the method includes determining a serum soluble B-cell maturation antigen (sBCMA) level in a sample obtained from a subject with MM. In some embodiments, based on the serum sBCMA level, the subject is selected for administration of T cell therapy and / or debulking of MM. In some embodiments, based on the serum sBCMA level, the subject is selected for administration of T cell therapy. In some embodiments, based on the serum sBCMA level, the subject is selected for debulking of MM. In some embodiments, based on the serum sBCMA level, the subject is selected for administration of T cell therapy and debulking of MM.

[0096] In some embodiments, the method includes determining whether the subject has a serum sBCMA level below about 600 ng / mL, e.g., below about 566 ng / mL or 500 ng / mL. In some embodiments, if the subject is determined to have a serum sBCMA level below about 600 ng / mL, e.g., below about 566 ng / mL or 500 ng / mL, the subject is administered a T cell therapy comprising a dose of genetically engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells). In some embodiments, if the subject is determined to have a serum sBCMA level below about 600 ng / mL, e.g., below about 566 ng / mL or 500 ng / mL, the subject is selected for administration of a T cell therapy comprising a dose of genetically engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells).

[0097] In some embodiments, the method includes determining whether the subject has a serum sBCMA level greater than about 600 ng / mL, e.g., greater than about 500 ng / mL or 566 ng / mL. In some embodiments, if the subject is determined to have a serum sBCMA level greater than about 600 ng / mL, e.g., greater than about 500 ng / mL or 566 ng / mL, the subject is administered a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells).

[0098] Thus, in some embodiments, a method comprises: (a) determining that a subject has a serum soluble B-cell maturation antigen (sBCMA) level below about 600 ng / ml; and (b) administering to the subject a T cell therapy comprising a dose of genetically engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method comprises administering to the subject a T cell therapy comprising a dose of genetically engineered T cells that express a chimeric antigen receptor (CAR) that binds B-cell maturation antigen (BCMA), wherein the subject has previously been determined to have a serum soluble B-cell maturation antigen (sBCMA) level below about 600 ng / ml. In some embodiments, the method comprises selecting a subject with multiple myeloma (MM) for treatment with a T cell therapy comprising a dose of genetically engineered T cells, comprising determining that the subject has a serum soluble B-cell maturation antigen (sBCMA) level below about 600 ng / ml, wherein if the subject is determined to have a serum sBCMA level below about 600 ng / ml, the subject is selected for administration with the T cell therapy.

[0099] In some embodiments, the method includes determining whether the subject has a serum sBCMA level greater than about 600 ng / mL, e.g., greater than about 500 ng / mL or 566 ng / mL. In some embodiments, if the subject is determined to have a serum sBCMA level greater than about 600 ng / mL, e.g., greater than about 500 ng / mL or 566 ng / mL, the multiple myeloma is debulked prior to administration of a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells). In some embodiments, the method includes determining whether the subject has a serum sBCMA level greater than about 600 ng / mL, e.g., greater than about 500 ng / mL or 566 ng / mL. In some embodiments, if the subject is determined to have a serum sBCMA level greater than about 600 ng / mL, e.g., greater than about 500 ng / mL or 566 ng / mL, the subject is selected for debulking of the multiple myeloma prior to administration of a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells).

[0100] Thus, in some embodiments, the method comprises: (a) determining that the subject has a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / ml; (b) debulking the MM; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method comprises administering to the subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein: (a) the subject was previously determined to have a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / ml; and (b) the MM was previously debulked between (1) the subject was determined to have a serum sBCMA level greater than about 600 ng / mL and (2) the subject was administered the T cell therapy. In some embodiments, the method includes determining that the subject has a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL, and if the subject is determined to have a serum sBCMA level greater than about 600 ng / mL, the subject is selected for debulking of MM prior to administering to the subject a T cell therapy comprising a dose of genetically engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method includes (a) determining that the subject has a first serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL; (b) debulking the MM; (c) determining that the subject has a post-debulking serum sBCMA level less than about 600 ng / mL; and (d) administering to the subject a T cell therapy comprising a dose of genetically engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA.

[0101] In some embodiments, debulking is performed as described in Section II.B.

[0102] In some embodiments, debulking comprises administering chemotherapy, radiation, or an immunomodulatory agent to the subject. In some embodiments, debulking comprises administering chemotherapy to the subject. In some embodiments, the chemotherapy comprises melphalan, doxorubicin, or cyclophosphamide chemotherapy. In some embodiments, debulking comprises administering radiation to the subject. In some embodiments, the immunomodulatory agent is thalidomide, lenalidomide, or pomalidomide. In some embodiments, it comprises administering an immunomodulatory agent to the subject. In some embodiments, the immunomodulatory agent is a checkpoint inhibitor.

[0103] In some embodiments, debulking is performed within about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week prior to administering T cell therapy to the subject. In some embodiments, debulking is performed within about 3 months prior to administering T cell therapy to the subject. In some embodiments, debulking is performed within about 2 months prior to administering T cell therapy to the subject. In some embodiments, debulking is performed within about 1 month prior to administering T cell therapy to the subject. In some embodiments, debulking is performed within about 3 weeks prior to administering T cell therapy to the subject. In some embodiments, debulking is performed within about 2 weeks prior to administering T cell therapy to the subject. In some embodiments, debulking is performed within about 1 week prior to administering T cell therapy to the subject. In some embodiments, debulking is performed prior to administering lymphodepleting therapy to the subject. In some embodiments, debulking is performed after administering lymphodepleting therapy to the subject. In some embodiments, the subject is treated with a gamma secretase inhibitor prior to administration of the T cell therapy to the subject.

[0104] In some embodiments, the method comprises determining that the subject has a serum sBCMA level greater than or less than about 590 ng / ml, about 580 ng / ml, about 570 ng / ml, about 560 ng / ml, about 550 ng / ml, about 540 ng / ml, about 530 ng / ml, about 520 ng / ml, about 510 ng / ml, or about 500 ng / ml. In some embodiments, the method comprises determining that the subject has a serum sBCMA level greater than or less than about 590 ng / ml. In some embodiments, the method comprises determining that the subject has a serum sBCMA level greater than or less than about 580 ng / ml. In some embodiments, the method comprises determining that the subject has a serum sBCMA level greater than or less than about 570 ng / ml. In some embodiments, the method comprises determining that the subject has a serum sBCMA level greater than or less than about 566 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 560 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 550 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 540 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 530 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 520 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 510 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 500 ng / ml.

[0105] In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 590 ng / ml, about 580 ng / ml, about 570 ng / ml, about 560 ng / ml, about 550 ng / ml, about 540 ng / ml, about 530 ng / ml, about 520 ng / ml, about 510 ng / ml, or about 500 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 590 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 580 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 570 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 566 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 560 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 550 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 540 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 530 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 520 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 510 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 500 ng / ml.

[0106] ii. Response In some embodiments, the method includes determining whether the subject has a serum soluble B-cell maturation antigen (sBCMA) level greater than or less than about 600 ng / mL. In some embodiments, the method includes determining the level of serum sBCMA in a sample obtained from the subject. In some embodiments, the subject is predicted to achieve CR or sCR after administration of the T cell therapy based on the level of serum sBCMA being less than a particular threshold, for example, 566 or 600 ng / mL.

[0107] In some embodiments, the method includes determining whether the subject has a serum sBCMA level below about 600 ng / mL, e.g., below about 566 ng / mL or 500 ng / mL. In some embodiments, if the subject is determined to have a serum sBCMA level below about 600 ng / mL, e.g., below about 566 ng / mL or 500 ng / mL, the subject is predicted to exhibit CR or sCR after administration of a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells). In some embodiments, if the subject is determined to have a serum sBCMA level below about 600 ng / mL, the subject is about 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or about 10-fold more likely to achieve CR or sCR after administration of T cell therapy compared to a subject with a serum sBCMA level above about 600 ng / mL and administered T cell therapy. In some embodiments, if a subject is determined to have a serum sBCMA level below about 600 ng / mL, the subject is about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% more likely to achieve a CR or sCR after administration of T cell therapy compared to a subject with a serum sBCMA level above about 600 ng / mL and who has been administered T cell therapy. In some embodiments, if a subject is determined to have a serum sBCMA level below about 566 ng / mL, the subject is about 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or about 10-fold more likely to achieve a CR or sCR after administration of T cell therapy compared to a subject with a serum sBCMA level above about 566 ng / mL and who has been administered T cell therapy. In some embodiments, if a subject is determined to have a serum sBCMA level below about 566 ng / mL, the subject is about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% more likely to achieve a CR or sCR after administration of the T cell therapy compared to a subject with a serum sBCMA level above about 566 ng / mL who was administered the T cell therapy.In some embodiments, if a subject is determined to have a serum sBCMA level below about 500 ng / mL, the subject is about 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or about 10-fold more likely to achieve a CR or sCR after administration of the T cell therapy compared to a subject with a serum sBCMA level above about 500 ng / mL who was administered the T cell therapy.

[0108] In some embodiments, the method includes determining whether the subject has a serum sBCMA level greater than about 600 ng / mL, e.g., greater than about 500 ng / mL or 566 ng / mL. In some embodiments, if the subject is determined to have a serum sBCMA level greater than about 600 ng / mL, e.g., greater than about 500 ng / mL or 566 ng / mL, the subject is predicted to not exhibit CR or sCR after administration of a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells). In some embodiments, if the subject is determined to have a serum sBCMA level greater than about 600 ng / mL, the subject is about 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or about 10-fold more likely to not achieve CR or sCR after administration of T cell therapy compared to a subject with a serum sBCMA level less than about 600 ng / mL and administered T cell therapy. In some embodiments, if a subject is determined to have a serum sBCMA level greater than about 600 ng / mL, the subject is about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% more likely to not achieve a CR or sCR after administration of T cell therapy compared to a subject with a serum sBCMA level less than about 600 ng / mL and administered T cell therapy. In some embodiments, if a subject is determined to have a serum sBCMA level greater than about 566 ng / mL, the subject is about 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or about 10-fold more likely to not achieve a CR or sCR after administration of T cell therapy compared to a subject with a serum sBCMA level less than about 566 ng / mL and administered T cell therapy. In some embodiments, if a subject is determined to have a serum sBCMA level greater than about 566 ng / mL, the subject is about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% more likely to not achieve a CR or sCR after administration of the T cell therapy compared to a subject with a serum sBCMA level less than about 566 ng / mL who was administered the T cell therapy.In some embodiments, if a subject is determined to have a serum sBCMA level greater than about 500 ng / mL, the subject is about 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or about 10-fold more likely to not achieve a CR or sCR after administration of the T cell therapy compared to a subject with a serum sBCMA level less than about 500 ng / mL and who has been administered the T cell therapy. In some embodiments, if a subject is determined to have a serum sBCMA level greater than about 500 ng / mL, the subject is about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% more likely to not achieve a CR or sCR after administration of the T cell therapy compared to a subject with a serum sBCMA level less than about 500 ng / mL and who has been administered the T cell therapy.

[0109] Thus, in some embodiments, a method includes predicting a response of a subject with multiple myeloma (MM) to treatment with a T cell therapy comprising a dose of engineered T cells, comprising determining that the subject has a serum soluble B cell maturation antigen (sBCMA) level below about 600 ng / ml, wherein if the subject has a serum sBCMA level below about 600 ng / ml, the subject is predicted to achieve a complete response (CR) or a stringent complete response (sCR).

[0110] In some embodiments, the method comprises determining that the subject has a serum sBCMA level greater than or less than about 590 ng / ml, about 580 ng / ml, about 570 ng / ml, about 560 ng / ml, about 550 ng / ml, about 540 ng / ml, about 530 ng / ml, about 520 ng / ml, about 510 ng / ml, or about 500 ng / ml. In some embodiments, the method comprises determining that the subject has a serum sBCMA level greater than or less than about 590 ng / ml. In some embodiments, the method comprises determining that the subject has a serum sBCMA level greater than or less than about 580 ng / ml. In some embodiments, the method comprises determining that the subject has a serum sBCMA level greater than or less than about 570 ng / ml. In some embodiments, the method comprises determining that the subject has a serum sBCMA level greater than or less than about 566 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 560 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 550 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 540 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 530 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 520 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 510 ng / ml. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than or less than about 500 ng / ml.

[0111] In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 590 ng / ml, about 580 ng / ml, about 570 ng / ml, about 560 ng / ml, about 550 ng / ml, about 540 ng / ml, about 530 ng / ml, about 520 ng / ml, about 510 ng / ml, or about 500 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 590 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 580 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 570 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 566 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 560 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 550 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 540 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 530 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 520 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 510 ng / ml. In some embodiments, the subject is determined to have a serum sBCMA level greater than or less than about 500 ng / ml.

[0112] B. Heavy chain disease i. Choices regarding treatment and / or debulking In some embodiments, the method includes determining the presence or absence of IgG heavy chain disease (HCD) in a sample obtained from the subject. In some embodiments, based on the presence or absence of IgG HCD, the subject is selected for administration of T cell therapy and / or debulking of MM. In some embodiments, based on the presence or absence of IgG HCD, the subject is selected for administration of T cell therapy. In some embodiments, based on the presence or absence of IgG HCD, the subject is selected for debulking of MM. In some embodiments, based on the presence or absence of IgG HCD, the subject is selected for administration of T cell therapy and debulking of MM.

[0113] In some embodiments, the method includes determining whether the subject does not have the presence of IgG HCD. In some embodiments, if the subject is determined to not have heavy chain disease, the subject is administered a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells). In some embodiments, if the subject is determined to not have heavy chain disease, the subject is selected for administration of a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells).

[0114] In some embodiments, the methods include determining whether the subject has the presence of IgG HCD. In some embodiments, if the subject is determined to have the presence of IgG HCD, the subject is not administered a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells).

[0115] Thus, in some embodiments, a method comprises: (a) determining that a subject does not have the presence of IgG heavy chain disease (HCD); and (b) administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method comprises administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds B-cell maturation antigen (BCMA), wherein the subject was previously determined to not have the presence of IgG heavy chain disease (HCD). In some embodiments, the method comprises selecting a subject with multiple myeloma (MM) for treatment with a T cell therapy comprising a dose of engineered T cells, wherein the subject is determined to not have the presence of IgG heavy chain disease (HCD), and the subject is selected for administration with the T cell therapy if the subject is determined to not have the presence of IgG HCD.

[0116] In some embodiments, the method includes determining whether the subject has the presence of IgG HCD. In some embodiments, if the subject is determined to have the presence of IgG HCD, the multiple myeloma is debulked prior to administration of T cell therapy comprising a dose of genetically engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells). In some embodiments, if the subject is determined to have the presence of IgG HCD, the subject is selected for debulking of the multiple myeloma prior to administration of T cell therapy comprising a dose of genetically engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells). In some embodiments, debulking is performed as described in the above section or in Section II.B.

[0117] In some embodiments, debulking comprises administering chemotherapy, radiation, or an immunomodulatory agent to the subject. In some embodiments, debulking comprises administering chemotherapy to the subject. In some embodiments, the chemotherapy comprises melphalan, doxorubicin, or cyclophosphamide chemotherapy. In some embodiments, debulking comprises administering radiation to the subject. In some embodiments, the immunomodulatory agent is thalidomide, lenalidomide, or pomalidomide. In some embodiments, it comprises administering an immunomodulatory agent to the subject. In some embodiments, the immunomodulatory agent is a checkpoint inhibitor.

[0118] In some embodiments, debulking is performed within about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week prior to administering T cell therapy to the subject. In some embodiments, debulking is performed within about 3 months prior to administering T cell therapy to the subject. In some embodiments, debulking is performed within about 2 months prior to administering T cell therapy to the subject. In some embodiments, debulking is performed within about 1 month prior to administering T cell therapy to the subject. In some embodiments, debulking is performed within about 3 weeks prior to administering T cell therapy to the subject. In some embodiments, debulking is performed within about 2 weeks prior to administering T cell therapy to the subject. In some embodiments, debulking is performed within about 1 week prior to administering T cell therapy to the subject. In some embodiments, debulking is performed prior to administering lymphodepleting therapy to the subject. In some embodiments, debulking is performed after administering lymphodepleting therapy to the subject. In some embodiments, the subject is treated with a gamma secretase inhibitor prior to administration of the T cell therapy to the subject.

[0119] ii. Response In some embodiments, the method includes determining that the subject does not have the presence of IgG heavy chain disease (HCD). In some embodiments, the method includes determining the presence or absence of IgG heavy chain disease (HCD) in a sample obtained from the subject. In some embodiments, based on the absence of IgG HCD, the subject is predicted to achieve CR or sCR after administration of T cell therapy. Conversely, in some embodiments, based on the presence of IgG HCD, the subject is predicted to not achieve CR or sCR after administration of T cell therapy.

[0120] In some embodiments, the method includes determining whether the subject does not have the presence of an IgG HCD. In some embodiments, if the subject is determined to not have the presence of an IgG HCD, the subject is predicted to exhibit CR or sCR after administration of a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells). In some embodiments, if the subject is determined to not have the presence of an IgG HCD, the subject is about 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or about 10-fold more likely to achieve CR or sCR after administration of the T cell therapy compared to a subject who has the presence of an IgG HCD and has been administered the T cell therapy.

[0121] In some embodiments, if a subject is determined to not have the presence of an IgG HCD, the subject is about 8-fold, 9-fold, 10-fold, 11-fold, or 12-fold more likely to achieve a CR or sCR after administration of T cell therapy compared to a subject who has the presence of an IgG HCD and has been administered T cell therapy. In some embodiments, a method includes determining whether a subject has the presence of an IgG HCD. In some embodiments, if a subject is determined to have the presence of an IgG HCD, the subject is predicted to not exhibit a CR or sCR after administration of T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA (i.e., BCMA CAR T cells). In some embodiments, if a subject is determined to have the presence of an IgG HCD, the subject is about 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or about 10-fold more likely to not achieve a CR or sCR after administration of T cell therapy compared to a subject who does not have the presence of an IgG HCD and has been administered T cell therapy. In some embodiments, if a subject is determined to have the presence of an IgG HCD, the subject is about 8, 9, 10, 11, or 12 times more likely to not achieve a CR or sCR after administration of the T cell therapy compared to a subject who does not have the presence of an IgG HCD and who has been administered the T cell therapy.

[0122] Thus, in some embodiments, a method includes predicting a response of a subject having multiple myeloma (MM) to treatment with a T cell therapy comprising a dose of genetically engineered cells, comprising determining that the subject does not have the presence of IgG heavy chain disease (HCD), where if the subject does not have the presence of IgG HCD, the subject is predicted to achieve a complete response (CR) or a stringent complete response (sCR). In some embodiments, a method includes predicting a response of a subject having multiple myeloma (MM) to treatment with a T cell therapy comprising a dose of genetically engineered cells, comprising determining that the subject has the presence of IgG heavy chain disease (HCD), where if the subject has the presence of IgG HCD, the subject is predicted not to achieve a complete response (CR) or a stringent complete response (sCR).

[0123] C. Other Biomarkers In some of any of the embodiments, the method includes assessing the copy number of the vector in the dose of engineered cells. In some embodiments, based on the copy number of the vector in the dose of engineered cells, the subject is predicted to achieve CR or sCR after administration of the T cell therapy, or is predicted not to achieve CR or sCR after administration of the T cell therapy.

[0124] In some embodiments, the method includes comparing the copy number of the vector in a dose of engineered cells to a threshold level, thereby determining the likelihood that the subject will achieve CR or sCR to T cell therapy. In some aspects, the threshold level is determined based on the mean or median copy number of the vector in a dose of engineered cells provided to a group of subjects, and a value within a range or standard deviation of the mean or median, wherein each subject in the group will or will not exhibit CR or sCR after receiving a dose of engineered cells. In some embodiments, if the copy number of the vector in the dose of engineered cells provided to a subject is lower than the threshold level, the subject is predicted to not exhibit CR or sCR to T cell therapy. In some embodiments, if the copy number of the vector in the dose of engineered cells provided to a subject is higher than the threshold level, the subject is predicted to exhibit CR or sCR to T cell therapy.

[0125] In some of any of the embodiments, the method includes assessing beta-2 microglobulin levels, such as in a sample obtained from the subject. In some embodiments, based on the beta-2 microglobulin level in the sample obtained from the subject, the subject is predicted to achieve CR or sCR after administration of the T cell therapy, or is predicted not to achieve CR or sCR after administration of the T cell therapy.

[0126] In some embodiments, the method includes individually comparing the level, amount, or concentration of beta-2 microglobulin from a sample obtained from the subject to a threshold level, thereby determining the likelihood that the subject will achieve CR or sCR to T cell therapy. In some aspects, the threshold level is determined based on the mean or median level, amount, or concentration of beta-2 microglobulin in biological samples obtained from a group of subjects before receiving T cell therapy, and values ​​within a range or standard deviation of the mean or median, wherein each of the subjects in the group will or will not achieve CR or sCR. In some embodiments, if the level, amount, or concentration of beta-2 microglobulin from a sample obtained from the subject is higher than the threshold level, the subject is predicted to not achieve CR or sCR to T cell therapy. In some embodiments, the threshold level is about 5.5 mg / mL. In some embodiments, if the level, amount, or concentration of beta-2 microglobulin from a sample obtained from the subject is higher than about 5.5 mg / mL, the subject is predicted to not achieve CR or sCR to T cell therapy. In some embodiments, if the level, amount, or concentration of beta-2 microglobulin from a sample obtained from the subject is lower than a threshold level, the subject is predicted to exhibit CR or sCR to T cell therapy. In some embodiments, the threshold level is about 3.5 mg / mL. In some embodiments, if the level, amount, or concentration of beta-2 microglobulin from a sample obtained from the subject is lower than about 3.5 mg / mL, the subject is predicted to exhibit CR or sCR to T cell therapy.

[0127] In some of the embodiments, the method includes assessing D-dimer levels, such as in a sample obtained from the subject. In some embodiments, the sample is a blood sample. In some embodiments, the method includes individually comparing the level, amount, or concentration of D-dimer from the sample obtained from the subject to a threshold level, thereby determining the likelihood that the subject will achieve CR or sCR to T cell therapy. In some aspects, the threshold level is determined based on the mean or median D-dimer level, amount, or concentration in biological samples obtained from a group of subjects before receiving T cell therapy, and values ​​within a range or standard deviation of the mean or median, and each of the subjects in the group will or will not exhibit CR or sCR. In some embodiments, if the level, amount, or concentration of D-dimer from the sample obtained from the subject is higher than the threshold level, it is predicted that the subject will not exhibit CR or sCR to T cell therapy. In some embodiments, the threshold level is about 0.8 mg / L, about 0.9 mg / L, about 1.0 mg / L, about 1.1 mg / L, about 1.2 mg / L, or about 1.3 mg / L. In some embodiments, if the level, amount, or concentration of D-dimer from a sample obtained from a subject is greater than about 0.8 mg / L, about 0.9 mg / L, about 1.0 mg / L, about 1.1 mg / L, about 1.2 mg / L, or about 1.3 mg / L, the subject is predicted not to exhibit CR or sCR to T cell therapy. In some embodiments, if the level, amount, or concentration of D-dimer from a sample obtained from a subject is less than the threshold level, the subject is predicted to exhibit CR or sCR to T cell therapy. In some embodiments, the threshold level is about 0.8 mg / L, about 0.7 mg / L, about 0.6 mg / L, about 0.5 mg / L, about 0.4 mg / L, or about 0.3 mg / L. In some embodiments, a subject is predicted to exhibit CR or sCR to the T cell therapy if the level, amount, or concentration of D-dimer from a sample obtained from the subject is less than about 0.8 mg / L, about 0.7 mg / L, about 0.6 mg / L, about 0.5 mg / L, about 0.4 mg / L, or about 0.3 mg / L.

[0128] In some of any of the embodiments, the method includes assessing ferritin levels, such as in a sample obtained from the subject. In some embodiments, based on the ferritin level in the sample obtained from the subject, the subject is predicted to achieve CR or sCR after administration of the T cell therapy, or is predicted not to achieve CR or sCR after administration of the T cell therapy.

[0129] In some embodiments, the method includes individually comparing the level, amount, or concentration of ferritin from a sample obtained from a subject with a threshold level, thereby determining the likelihood that the subject will achieve CR or sCR to T cell therapy. In some aspects, the threshold level is determined based on the mean or median ferritin level, amount, or concentration in biological samples obtained from a group of subjects before receiving T cell therapy, and values ​​within a range or standard deviation of the mean or median, where each subject in the group will or will not show CR or sCR. In some embodiments, if the level, amount, or concentration of ferritin from a sample obtained from a subject is higher than the threshold level, the subject is predicted not to show CR or sCR to T cell therapy. In some embodiments, the threshold level is about 350 μg / L, about 400 μg / L, about 450 μg / L, about 500 μg / L, about 550 μg / L, or about 600 μg / L. In some embodiments, if the level, amount, or concentration of ferritin from a sample obtained from the subject is greater than about 350 μg / L, about 400 μg / L, about 450 μg / L, about 500 μg / L, about 550 μg / L, or about 600 μg / L, the subject is predicted not to exhibit CR or sCR to T cell therapy. In some embodiments, if the level, amount, or concentration of ferritin from a sample obtained from the subject is less than a threshold level, the subject is predicted to exhibit CR or sCR to T cell therapy. In some embodiments, the threshold level is about 300 μg / L, about 250 μg / L, about 200 μg / L, about 150 μg / L, about 100 μg / L, or about 50 μg / L. In some embodiments, a subject is predicted to exhibit CR or sCR to the T cell therapy if the level, amount, or concentration of ferritin from a sample obtained from the subject is less than about 300 μg / L, about 250 μg / L, about 200 μg / L, about 150 μg / L, about 100 μg / L, or about 50 μg / L.

[0130] In some of any of the embodiments, the method includes assessing sodium levels, such as in a sample obtained from the subject. In some embodiments, based on the sodium level in the sample from the subject, the subject is predicted to achieve CR or sCR after administration of the T cell therapy, or is predicted not to achieve CR or sCR after administration of the T cell therapy.

[0131] In some embodiments, the method includes individually comparing the level, amount, or concentration of sodium from a sample obtained from the subject to a threshold level, thereby determining the likelihood that the subject will achieve CR or sCR to T cell therapy. In some aspects, the threshold level is determined based on the mean or median sodium level, amount, or concentration in biological samples obtained from a group of subjects prior to T cell therapy, and values ​​within a range or standard deviation of the mean or median, wherein each of the subjects in the group will or will not achieve CR or sCR. In some embodiments, if the sodium level, amount, or concentration from the sample obtained from the subject is lower than the threshold level, the subject is predicted not to achieve CR or sCR to T cell therapy. In some embodiments, the threshold level is about 140 mmol / L, about 139 mmol / L, about 138 mmol / L, about 137 mmol / L, about 136 mmol / L, or about 135 mmol / L. In some embodiments, if the level, amount, or concentration of sodium from a sample obtained from the subject is lower than about 140 mmol / L, about 139 mmol / L, about 138 mmol / L, about 137 mmol / L, about 136 mmol / L, or about 135 mmol / L, the subject is predicted not to exhibit CR or sCR to the T cell therapy. In some embodiments, if the level, amount, or concentration of sodium from a sample obtained from the subject is higher than a threshold level, the subject is predicted to exhibit CR or sCR to the T cell therapy. In some embodiments, the threshold level is about 140 mmol / L, about 141 mmol / L, about 142 mmol / L, about 143 mmol / L, about 144 mmol / L, or about 145 mmol / L. In some embodiments, the subject is predicted to exhibit CR or sCR to the T cell therapy if the level, amount, or concentration of sodium from a sample obtained from the subject is greater than about 140 mmol / L, about 141 mmol / L, about 142 mmol / L, about 143 mmol / L, about 144 mmol / L, or about 145 mmol / L. In some of any of the embodiments, the method includes assessing the prothrombin time-international normalized ratio (PT-INR), such as in the sample obtained from the subject. In some embodiments, the sample is a blood sample.In some embodiments, the method includes comparing the PT-INR from a sample obtained from the subject to a threshold PT-INR, thereby determining the likelihood that the subject will achieve CR or sCR to the T cell therapy. In some aspects, the threshold PT-INR is determined based on the mean or median PT-INR and values ​​within a range or standard deviation of the mean or median in biological samples obtained from a group of subjects before receiving T cell therapy, each of whom will or will not experience CR or sCR. In some embodiments, if the PT-INR of the sample obtained from the subject is higher than the threshold PT-INR, it is predicted that the subject will not experience CR or sCR to the T cell therapy. In some embodiments, if the PT-INR of a sample obtained from a subject is higher than the threshold PT-INR, the subject is about 100-fold, 150-fold, 200-fold, 250-fold, or 300-fold less likely to exhibit CR or sCR to T cell therapy compared to a subject whose sample has a PT-INR at the threshold PT-INR or a PT-INR below the threshold PT-INR. In some embodiments, if the PT-INR of a sample obtained from a subject is lower than the threshold PT-INR, the subject is predicted to exhibit CR or sCR to T cell therapy. In some embodiments, if the PT-INR of a sample obtained from a subject is lower than the threshold PT-INR, the subject is about 100-fold, 150-fold, 200-fold, 250-fold, or 300-fold more likely to exhibit CR or sCR to T cell therapy compared to a subject whose sample has a PT-INR at the threshold PT-INR or a PT-INR higher than the threshold PT-INR.

[0132] II. Methods of Treatment and Use Also provided herein are methods of and uses of the BCMA binding molecules, recombinant receptors, engineered cells, and pharmaceutical compositions and formulations thereof, such as in methods of treating, and / or detecting, selecting, diagnosing, and prognosing multiple myeloma. Some such methods, e.g., methods of treatment and use, include administering to a subject an engineered cell, e.g., a plurality of engineered cells (e.g., BCMA CAR T cells), that express a provided anti-BCMA recombinant receptor.

[0133] A. Target and Usage Also provided are methods and uses, e.g., therapeutic uses, of administering an anti-BCMA recombinant receptor (e.g., a CAR), an engineered cell expressing the recombinant receptor (e.g., a CAR), a plurality of engineered cells expressing the receptor, and / or a composition comprising the same. Such methods and uses include therapeutic methods and uses, for example, administering a molecule (e.g., a recombinant receptor), a cell (e.g., an engineered cell), or a composition containing the same to a subject with multiple myeloma (MM). In some embodiments, the molecule, cell, and / or composition is administered in an effective amount to provide treatment for MM. Provided herein are uses of recombinant receptors (e.g., a CAR) and cells (e.g., engineered cells) in such methods and treatments, as well as in the preparation of medicaments for carrying out such therapeutic methods. In some embodiments, the methods are carried out by administering the binding molecule or cell, or a composition comprising the same, to a subject with MM. In some embodiments, the methods thereby treat MM in the subject. Also provided herein are uses of any of the compositions, such as the pharmaceutical compositions provided herein, for the treatment of multiple myeloma (MM), such as for use in a treatment regimen.

[0134] As used herein, "treatment" (and grammatical variations thereof, such as "treat" or "treating") refers to the complete or partial remission or reduction of a disease or condition or disorder, or the symptoms, adverse effects or outcomes, or phenotypes associated therewith. Desirable effects of treatment include, but are not limited to, prevention of disease recurrence, alleviation of symptoms, reduction of any direct or indirect pathological consequences, prevention of metastasis, reduction in the rate of disease progression, remission or palliation of the disease state, and reduction or improvement in prognosis. The term does not imply a complete cure of the disease, or complete elimination of any symptoms, or an effect(s) on all symptoms or outcomes.

[0135] As used herein, "delaying the onset of disease" means to postpone, prevent, slow down, delay, stabilize, inhibit, and / or postpone the onset of a disease (e.g., cancer). This delay can be of varying lengths of time, depending on the disease being treated and / or the subject's medical history. A sufficient or significant delay can, in effect, encompass prevention, in that the subject does not develop the disease. For example, the onset of late-stage cancer, such as metastasis, can be delayed.

[0136] "Preventing," as used herein, includes providing prophylaxis against the occurrence or recurrence of a disease in a subject who may be predisposed to the disease but has not yet been diagnosed with the disease. In some embodiments, the molecules and compositions provided are used to delay the onset of the disease or slow the progression of the disease.

[0137] As used herein, "inhibiting" a function or activity means reducing the function or activity when compared to otherwise identical conditions, except for the condition or parameter of interest, or when compared to another condition. For example, an antibody or composition or cell that inhibits tumor growth reduces the rate of tumor growth compared to the rate of tumor growth in the absence of the antibody or composition or cell.

[0138] An "effective amount" of an agent, e.g., a pharmaceutical formulation, binding molecule, antibody, cell, or composition, refers to an amount effective, in the context of administration, at dosages / amounts and for periods of time necessary to achieve a desired result, e.g., a therapeutic or prophylactic result.

[0139] A "therapeutically effective amount" of an agent, e.g., a pharmaceutical formulation, binding molecule, antibody, cell, or composition, refers to an amount effective, at dosages and for periods of time necessary, to achieve a desired therapeutic result, e.g., treatment of a disease, condition, or disorder, and / or the pharmacokinetic or pharmacodynamic effects of the treatment. A therapeutically effective amount may vary according to factors such as the disease state, age, sex, and weight of the subject, and the population of cells administered. In some embodiments, provided methods involve administering a molecule, antibody, cell, and / or composition in an effective amount, e.g., a therapeutically effective amount.

[0140] A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically effective amount will usually, but not necessarily, be less than the therapeutically effective amount.

[0141] As used herein, a "subject" or "individual" is a human.

[0142] The method for administering cells for adoptive cell therapy is known, and can be used in conjunction with the provided method and composition.For example, the method of adoptive T cell therapy is described in, for example, Gruenberg et al., U.S. Patent Application Publication No. 2003 / 0170238; Rosenberg, U.S. Patent No. 4,690,915; Rosenberg (2011) Nat Rev Clin Oncol.8(10):577-85.For example, see Themeli et al. (2013) Nat Biotechnol.31(10):928-933; Tsukahara et al. (2013) Biochem Biophys Res Commun 438(1):84-9; Davila et al. (2013) PLoS ONE 8(4):e61338.

[0143] Among the diseases to be treated, multiple myeloma (MM) is associated with BCMA expression. For a review of BCMA, see Coquery et al., Crit Rev Immunol., 2012, 32(4):287-305. BCMA has been implicated in mediating tumor cell survival and is therefore a potential target for cancer therapy. Chimeric antigen receptors containing mouse anti-human BCMA antibodies and cells expressing such chimeric receptors have previously been described. See Carpenter et al., Clin Cancer Res., 2013, 19(8):2048-2060.

[0144] In some embodiments, the multiple myeloma (MM) is high-risk MM, or relapsed and / or refractory multiple myeloma. In some embodiments, the multiple myeloma (MM) is high-risk MM. In some embodiments, the high-risk MM includes IMWG high-risk cytogenetics. In some embodiments, at the time of administration of the cell therapy, the subject has IMWG high-risk cytogenetics. In some embodiments, the high-risk cytogenetics include del(17p), t(4:14), and t(14;16). In some embodiments, the multiple myeloma (MM) is relapsed and / or refractory multiple myeloma. In some embodiments, the multiple myeloma (MM) is relapsed and refractory multiple myeloma (r / r MM). In some embodiments, at the time of administration, the subject has R / R MM. In some embodiments, the method can identify subjects who have, are suspected of having, or are at risk of developing multiple myeloma. Accordingly, provided herein are methods for identifying subjects with multiple myeloma, selecting subjects for treatment with and / or administering to a subject any of the BCMA-binding recombinant receptors (e.g., CARs) described herein, or engineered cells expressing same.

[0145] In some embodiments, the subject has a serum soluble B-cell maturation antigen (sBCMA) level below about 600 ng / mL. In some embodiments, the method comprises determining that the subject has a serum sBCMA level below about 600 ng / mL. In some embodiments, the subject has a serum soluble B-cell maturation antigen (sBCMA) level below about 566 ng / mL. In some embodiments, the method comprises determining that the subject has a serum sBCMA level below about 566 ng / mL. In some embodiments, the subject has a serum soluble B-cell maturation antigen (sBCMA) level below about 500 ng / mL. In some embodiments, the method comprises determining that the subject has a serum sBCMA level below about 500 ng / mL. In some embodiments, the serum sBCMA level is determined prior to administration of lymphodepleting therapy to the subject, prior to leukapheresis, and / or prior to administration of T-cell therapy to the subject. In some embodiments, the serum sBCMA level is determined prior to administration of lymphodepleting therapy to the subject. In some embodiments, the determination of serum sBCMA levels is performed prior to leukapheresis. In some embodiments, serum sBCMA is determined prior to administration of T cell therapy to the subject. In some embodiments, methods are provided for selecting a subject for treatment with T cell therapy. In some embodiments, if a subject is determined to have a serum sBCMA level lower than about 600 ng / mL, the subject is selected for treatment. In some embodiments, if a subject is determined to have a serum sBCMA level higher than about 600 ng / mL, the subject is not selected for treatment. In some embodiments, if a subject is determined to have a serum sBCMA level lower than about 566 ng / mL, the subject is not selected for treatment. In some embodiments, if a subject is determined to have a serum sBCMA level higher than about 566 ng / mL, the subject is selected for treatment. In some embodiments, if a subject is determined to have a serum sBCMA level lower than about 500 ng / mL, the subject is selected for treatment. In some embodiments, if a subject is determined to have a serum sBCMA level higher than about 500 ng / mL, the subject is not selected for treatment.In some embodiments, if a subject is determined to have a serum sBCMA level greater than about 600 ng / mL, the subject is selected for debulking of multiple myeloma prior to administering T cell therapy to the subject. In some embodiments, if a subject is determined to have a serum sBCMA level greater than about 566 ng / mL, the subject is selected for debulking of multiple myeloma prior to administering T cell therapy to the subject. In some embodiments, if a subject is determined to have a serum sBCMA level greater than about 500 ng / mL, the subject is selected for debulking of multiple myeloma prior to administering T cell therapy to the subject.

[0146] In some embodiments, the subject does not have the presence of IgG heavy chain disease (HCD). In some embodiments, the method includes determining that the subject does not have the presence of IgG HCD. In some embodiments, the absence of IgG HCD is determined prior to administration of lymphodepleting therapy to the subject, prior to leukapheresis, and / or prior to administration of T cell therapy to the subject. In some embodiments, the absence of IgG HCD is determined prior to administration of lymphodepleting therapy to the subject. In some embodiments, the absence of IgG HCD is determined prior to leukapheresis. In some embodiments, the absence of IgG HCD is determined prior to administration of T cell therapy to the subject. In some embodiments, methods are provided for selecting a subject for treatment with T cell therapy. In some embodiments, if the subject is determined to not have the presence of IgG HCD, the subject is selected for treatment. In some embodiments, if the subject is determined to have the presence of IgG HCD, the subject is not selected for treatment. In some embodiments, if the subject is determined to have the presence of IgG HCD, the subject is selected for debulking of the multiple myeloma prior to administration of T cell therapy to the subject.

[0147] In some embodiments, the subject has persistent or recurrent disease following treatment with a prior line of therapy. In some embodiments, before administering T cell therapy, the subject has undergone one or more prior therapies. In some embodiments, the subject has undergone at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more prior therapies. In some embodiments, the subject has undergone at least 3, 4, 5, 6, 7, 8, 9, 10 or more prior therapies. In some embodiments, the subject has undergone three or more prior therapies.

[0148] In some embodiments, each of the three or more prior lines of therapy included two consecutive cycles. In some embodiments, each of the three or more prior lines of therapy included two consecutive cycles, unless progressive disease was the best response to that line of therapy. In some embodiments, progressive disease is progression within 60 days after the last dose of a line of therapy. In some embodiments, the subject is refractory to the last of the three or more prior lines of therapy. In some embodiments, the three or more prior lines of therapy include a proteasome inhibitor (PI), an immunomodulatory agent, and an anti-CD38 antibody.

[0149] In some of any of the embodiments, the immunomodulatory agent is selected from thalidomide, lenalidomide, or pomalidomide. In some of any of the embodiments, the proteasome inhibitor is selected from bortezomib, carfilzomib, and ixazomib. In some of any of the embodiments, the anti-CD38 antibody is or includes daratumumab.

[0150] In some embodiments, the criteria for measurable disease for multiple myeloma can include (1) serum M protein ≥ 1 g / dL; (2) urinary M protein ≥ 200 mg / 24 hours; and (3) diseased serum free light chain (sFLC) levels ≥ 10 mg / dL and an abnormal Kappa to λ ratio. In some cases, light chain disease is permitted only for subjects without measurable disease in serum or urine. In some embodiments, the subject has measurable disease at the time of administration of T cell therapy. In some embodiments, measurable disease includes (i) serum M protein ≥ 1.0 g / dL; (ii) urinary M protein ≥ 200 mg / 24 hours; and / or (iii) if the serum free light chain (FLC) ratio is abnormal, diseased serum FLC levels ≥ 10 mg / dL. In some embodiments, measurable disease includes serum M protein ≥ 1.0 g / dL. In some embodiments, measurable disease includes urinary M protein ≥ 200 mg / 24 hours. In some embodiments, measurable disease includes, when the serum free light chain (FLC) ratio is abnormal, a diseased serum FLC level of 10 mg / dL or greater. In some embodiments, measurable disease includes (i) serum M protein 1.0 g or greater; (ii) urinary M protein 200 mg or greater / 24 hours; and (iii) when the serum free light chain (FLC) ratio is abnormal, a diseased serum FLC level of 10 mg / dL or greater. See Kumar et al., Lancet Oncol (2016) 17(8):e328-46.

[0151] In some embodiments, the subject has adequate organ function.

[0152] In some embodiments, the subject is 18 years of age or older.

[0153] In some embodiments, the method can involve including or excluding certain subjects for treatment with T cell therapy based on certain criteria, diagnoses, or indicators. In some embodiments, at the time of administration of T cell therapy or pre-treatment lymphodepleting chemotherapy, the subject has no known central nervous system (CNS) involvement with myeloma. In some embodiments, at the time of administration of T cell therapy or pre-treatment lymphodepleting chemotherapy, the subject has no history or presence of clinically relevant CNS pathology. In some embodiments, at the time of administration of T cell therapy or pre-treatment lymphodepleting chemotherapy, the subject has no active or history of plasma cell leukemia (PCL). In some embodiments, at the time of administration of T cell therapy or pre-treatment lymphodepleting chemotherapy, the subject does not have solitary plasmacytoma or non-secretory myeloma without other evidence of measurable disease. In some embodiments, at the time of administration of T cell therapy or pre-treatment lymphodepleting chemotherapy, the subject has no history of allogeneic hematopoietic stem cell transplantation. In some embodiments, at the time of administration of the T cell therapy or pre-treatment lymphodepleting chemotherapy, the subject has not previously been treated with a gene therapy-based therapeutic for cancer. In some embodiments, at the time of administration of the T cell therapy or pre-treatment lymphodepleting chemotherapy, the subject has not previously been treated with an investigational cell therapy for cancer. In some embodiments, at the time of administration of the T cell therapy or pre-treatment lymphodepleting chemotherapy, the subject has not previously been treated with a BCMA-targeted therapy.

[0154] In some embodiments, evaluation of the criteria, diagnostics, or indicators can be performed at the time of screening a subject for eligibility or suitability for treatment according to the provided methods, at various steps of a treatment regimen, at the time of receiving lymphodepletion therapy, and / or at or immediately prior to the initiation of administration of engineered cells or compositions thereof.

[0155] Thus, the provided methods and uses include methods and uses for adoptive cell therapy. In some embodiments, the methods include administering cells or compositions containing cells to a subject, tissue, or cell, e.g., one having, at risk for, or suspected of having multiple myeloma. In some embodiments, the cells, populations, and compositions are administered to a subject having multiple myeloma, e.g., by adoptive cell therapy, e.g., adoptive T cell therapy. In some embodiments, the cells or compositions are administered to a subject, e.g., a subject having or at risk for multiple myeloma. In some aspects, the methods thereby treat, e.g., ameliorate, one or more symptoms of multiple myeloma, such as by reducing tumor burden.

[0156] The method for administering cells for adoptive cell therapy is known, and can be used in conjunction with the provided method and composition.For example, the method of adoptive T cell therapy is described in, for example, Gruenberg et al., U.S. Patent Application Publication No. 2003 / 0170238; Rosenberg, U.S. Patent No. 4,690,915; Rosenberg (2011) Nat Rev Clin Oncol.8(10):577-85.For example, see Themeli et al. (2013) Nat Biotechnol.31(10):928-933; Tsukahara et al. (2013) Biochem Biophys Res Commun 438(1):84-9; Davila et al. (2013) PLoS ONE 8(4):e61338.

[0157] In some embodiments, T cell therapy, e.g., adoptive cell therapy, e.g., adoptive T cell therapy, is carried out by autologous transfer, whereby cells are isolated and / or otherwise prepared from a subject to receive T cell therapy or from a sample derived from such a subject. Thus, in some aspects, the cells are derived from a subject, e.g., a patient, in need of treatment, and the cells, after isolation and processing, are administered to the same subject.

[0158] In some embodiments, T cell therapy, e.g., adoptive cell therapy, e.g., adoptive T cell therapy, is performed by allogeneic transfer, whereby cells are isolated and / or otherwise prepared from a subject other than the subject that is to or will ultimately receive cell therapy, e.g., a first subject. In such embodiments, the cells are then administered to a different subject of the same species, e.g., a second subject. In some embodiments, the first and second subjects are genetically identical. In some embodiments, the first and second subjects are genetically similar. In some embodiments, the second subject expresses the same HLA class or supertype as the first subject.

[0159] The subject may be male or female and of any suitable age, including infant, juvenile, adolescent, adult, and geriatric subjects, hi some embodiments, the subject is an adult (18 years of age or older).

[0160] In some embodiments, the dose and / or administration frequency are determined based on efficacy and / or response. In some embodiments, efficacy is determined by assessing the disease state. Exemplary methods for assessing the disease state include measuring M protein in biological fluids such as blood and / or urine by electrophoresis and immunofixation; quantifying sFLC (κ and λ) in the blood; skeletal examination; and imaging by positron emission tomography (PET) / computed tomography (CT) in subjects with extramedullary disease. In some embodiments, the disease state can be assessed by bone marrow examination. In some examples, the dose and / or administration frequency are determined by the expansion and persistence of the recombinant receptor or cells in the blood and / or bone marrow. In some embodiments, the dose and / or administration frequency are determined based on the anti-tumor activity of the recombinant receptor or engineered cells. In some embodiments, anti-tumor activity is determined by overall response rate (ORR) and / or International Myeloma Working Group (IMWG) Unified Response Criteria (see Kumar et al. (2016) Lancet Oncol 17(8):e328-346). In some embodiments, response is assessed using minimal residual disease (MRD) assessment. In some embodiments, MRD can be assessed by methods such as flow cytometry and high-throughput sequencing, e.g., deep sequencing. In some embodiments, response is assessed based on the duration of response following administration of the recombinant receptor or cells. In some examples, the dose and / or dosing frequency can be based on toxicity. In some embodiments, the dose and / or frequency can be determined based on the health-related quality of life (HRQoL) of the subject to whom the recombinant receptor and / or cells are administered. In some embodiments, the dose and / or dosing frequency can be altered, i.e., increased or decreased, based on any of the above criteria.

[0161] In some embodiments, the Eastern Cooperative Oncology Group (ECOG) performance status index can be used to evaluate or select subjects for treatment, for example, subjects who have had poor results from previous therapy (see, e.g., Oken et al. (1982) Am J Clin Oncol. 5:649-655). The ECOG performance status scale describes a patient's level of function in terms of their ability to manage themselves, their daily activities, and their physical abilities (e.g., walking, working, etc.). In some embodiments, an ECOG performance status of 0 indicates that the subject can perform normal activities. In some aspects, a subject with an ECOG performance status of 1 shows some limitations in physical activity, but the subject is fully ambulatory. In some aspects, a patient with an ECOG performance status of 2 is more than 50% ambulatory. In some cases, a subject with an ECOG performance status of 2 can also be capable of self-care; see, e.g., Sorensen et al., (1993) Br J Cancer 67(4) 773-775. In some embodiments, subjects to be administered according to the methods or treatment regimens provided herein include subjects with an ECOG performance status of 0 or 1. In some embodiments, the subject has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1. In some embodiments, the subject has an Eastern Cooperative Oncology Group (ECOG) performance status of 0. In some embodiments, the subject has an Eastern Cooperative Oncology Group (ECOG) performance status of 1.

[0162] In some embodiments, administration can treat a subject despite the subject becoming refractory to another therapy. In some embodiments, when administered to a subject in accordance with the embodiments described herein, a dose or composition can achieve a stringent complete response (sCR) or complete response (CR) in at least 20%, 30%, 40%, 50%, 60%, or 70% of the subjects to which it is administered. In some embodiments, when administered to a subject in accordance with the embodiments described herein, a dose or composition can achieve a stringent complete response (sCR) in at least 20%, 30%, 40%, 50%, 60%, or 70% of the subjects to which it is administered. In some embodiments, when administered to a subject in accordance with the embodiments described herein, a dose or composition can achieve a complete response (CR) in at least 20%, 30%, 40%, 50%, 60%, or 70% of the subjects to which it is administered. In some embodiments, for example, a particular response to treatment according to the methods provided herein can be assessed based on the International Myeloma Working Group (IMWG) Uniform Response Criteria (see Kumar et al. (2016) Lancet Oncol 17(8):e328-346).

[0163] In some embodiments, toxicity and / or side effects of treatment can be monitored and used to adjust the dose and / or administration frequency of the recombinant receptor, e.g., CAR, cell, and / or composition. For example, adverse events and abnormal laboratory test values ​​can be monitored and used to adjust the dose and / or administration frequency. Adverse events include infusion reactions, cytokine release syndrome (CRS), neurotoxicity, macrophage activation syndrome, and tumor lysis syndrome (TLS). Any of these events may establish dose-limiting toxicity and warrant dose reduction and / or discontinuation of treatment. Other side effects or adverse events that can be used as guidelines for establishing dose and / or dosing frequency include non-hematological adverse events, including fatigue, fever or febrile neutropenia, elevated transaminases for a period of time (e.g., 2 weeks or less or 7 days or less), headache, bone pain, hypotension, hypoxia, chills, diarrhea, nausea / vomiting, neurotoxicity (e.g., confusion, aphasia, seizures, lethargy, and / or abnormal mental status), disseminated intravascular coagulation, other asymptomatic non-hematological laboratory abnormalities, such as electrolyte abnormalities. Other side effects or adverse events that can be used as guidelines for establishing dose and / or dosing frequency include hematological adverse events, including, but not limited to, neutropenia, leukopenia, thrombocytopenia, thyroid and / or B-cell aplasia, and hypogammaglobinemia.

[0164] In some embodiments, treatment in accordance with the provided methods may result in a lower rate of toxicity and / or a lower degree of toxicity, toxic outcomes or symptoms, toxicity-promoting profiles, factors, or characteristics, e.g., symptoms or outcomes associated with or indicative of cytokine release syndrome (CRS) or neurotoxicity, e.g., severe CRS or severe neurotoxicity, e.g., compared to administration of other therapies.

[0165] In some embodiments, subject can receive bridging therapy after leukapheresis and before lymphodepletion chemotherapy.The treating physician can determine whether bridging therapy is necessary, for example, for disease control, during the preparation of the provided composition or cell.In some embodiments, bridging therapy is stopped before lymphodepletion begins.In some embodiments, bridging therapy is stopped 1 day, 2 days, 3 days, 4 days, 5 days, 7 days, 10 days, 14 days, 21 days, 28 days, 45 days or 60 days before lymphodepletion.

[0166] Once the cells are administered to a mammal (e.g., a human), in some aspects, the biological activity of the engineered cell population and / or antibody is measured by any of a number of known methods. Parameters for evaluation include specific binding of engineered or natural T cells or other immune cells to an antigen in vivo, e.g., by imaging, or ex vivo, e.g., by ELISA or flow cytometry. In certain embodiments, the ability of the engineered cells to destroy target cells can be measured using any suitable method known in the art, such as the cytotoxicity assays described in Kochenderfer et al., J. Immunotherapy, 32(7): 689-702 (2009) and Herman et al. J. Immunological Methods, 285(1): 25-40 (2004). In certain embodiments, the biological activity of the cells is measured by assaying the expression and / or secretion of certain cytokines, such as CD107a, IFNγ, IL-2, and TNF. In some aspects, biological activity is measured by assessing a clinical outcome, such as reduction in tumor burden or burden.

[0167] In certain embodiments, engineered cells are modified in several ways to increase their therapeutic or preventive efficacy.For example, in some embodiments, the engineered CAR expressed by cells is directly or indirectly conjugated to targeting moiety via linker.The solid line of compound (for example, CAR) conjugated to targeting moiety is known in the art.For example, see Wadwa et al., J. Drug Targeting, 3(2):111 (1995) and U.S. Patent No. 5,087,616.

[0168] B. Tumor debulking In some embodiments of the methods provided herein, the methods include debulking multiple myeloma (MM) prior to administering T cell therapy to the subject. In some embodiments of the methods provided herein, the methods include selecting a subject for debulking MM prior to administering T cell therapy to the subject.

[0169] In some embodiments, the method comprises determining that the subject has the presence of IgG heavy chain disease (HCD) and / or has a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL. In some embodiments, the method comprises determining that the subject has the presence of IgG heavy chain disease (HCD) and / or has a serum soluble B-cell maturation antigen (sBCMA) level greater than about 566 ng / mL. In some embodiments, the method comprises determining that the subject has the presence of IgG heavy chain disease (HCD) and / or has a serum soluble B-cell maturation antigen (sBCMA) level greater than about 500 ng / mL. In some embodiments, the subject has been determined to have the presence of IgG HCD and / or has a serum sBCMA level greater than about 600 ng / mL. In some embodiments, the subject has been determined to have the presence of IgG HCD and / or has a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the subject has been determined to have the presence of IgG HCD and / or has serum sBCMA levels greater than about 500 ng / mL.

[0170] In some embodiments, the method comprises determining that the subject has a serum sBCMA level greater than or less than about 590 ng / mL, about 580 ng / mL, about 570 ng / mL, about 560 ng / mL, about 550 ng / mL, about 540 ng / mL, about 530 ng / mL, about 520 ng / mL, about 510 ng / mL, or about 500 ng / mL. In some embodiments, the subject is determined to have a serum sBCMA level greater than about 590 ng / mL, about 580 ng / mL, about 570 ng / mL, about 560 ng / mL, about 550 ng / mL, about 540 ng / mL, about 530 ng / mL, about 520 ng / mL, about 510 ng / mL, or about 500 ng / mL. In some embodiments, the method comprises determining that the subject has a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the subject is determined to have a serum sBCMA level greater than about 566 ng / mL.

[0171] In some embodiments, the method includes debulking the MM when the subject is determined to have the presence of IgG HCD and / or have a serum sBCMA level greater than about 600 ng / mL. In some embodiments, the method includes debulking the MM when the subject is determined to have the presence of IgG HCD and / or have a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the method includes debulking the MM when the subject is determined to have the presence of IgG HCD and / or have a serum sBCMA level greater than about 500 ng / mL. In some embodiments, the MM is debulked when the subject is determined to have the presence of IgG HCD and / or have a serum sBCMA level greater than about 600 ng / mL. In some embodiments, the MM is debulked when the subject is determined to have the presence of IgG HCD and / or have a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the MM is debulked if the subject is determined to have the presence of IgG HCD and / or a serum sBCMA level greater than about 500 ng / mL. In some embodiments, the subject is selected for debulking if the subject is determined to have the presence of IgG HCD and / or a serum sBCMA level greater than about 600 ng / mL. In some embodiments, the subject is selected for debulking if the subject is determined to have the presence of IgG HCD and / or a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the subject is selected for debulking if the subject is determined to have the presence of IgG HCD and / or a serum sBCMA level greater than about 500 ng / mL.

[0172] In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than about 600 ng / mL. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the method includes determining that the subject has a serum sBCMA level greater than about 500 ng / mL. In some embodiments, the subject has been determined to have a serum sBCMA level greater than about 600 ng / mL. In some embodiments, the subject has been determined to have a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the subject has been determined to have a serum sBCMA level greater than about 500 ng / mL. In some embodiments, if the subject is determined to have a serum sBCMA level greater than about 600 ng / mL, the method includes debulking the MM. In some embodiments, if the subject is determined to have a serum sBCMA level greater than about 566 ng / mL, the method includes debulking the MM. In some embodiments, the method includes debulking the MM if the subject is determined to have a serum sBCMA level greater than about 500 ng / mL. In some embodiments, the MM is debulked if the subject is determined to have a serum sBCMA level greater than about 600 ng / mL. In some embodiments, the MM is debulked if the subject is determined to have a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the MM is debulked if the subject is determined to have a serum sBCMA level greater than about 500 ng / mL. In some embodiments, the subject is selected for debulking if the subject is determined to have a serum sBCMA level greater than about 600 ng / mL. In some embodiments, the subject is selected for debulking if the subject is determined to have a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the subject is selected for debulking if the subject is determined to have a serum sBCMA level greater than about 500 ng / mL.

[0173] In some embodiments, the method includes determining that the subject has the presence of IgG HCD. In some embodiments, the subject has been determined to have the presence of IgG HCD. In some embodiments, if the subject is determined to have the presence of IgG HCD, the method includes debulking the MM. In some embodiments, if the subject is determined to have the presence of IgG HCD, the MM is debulked. In some embodiments, if the subject is determined to have the presence of IgG HCD, the subject is selected for debulking.

[0174] In some embodiments, the method includes determining that the subject has the presence of IgG HCD and has a serum sBCMA level greater than about 600 ng / mL. In some embodiments, the method includes determining that the subject has the presence of IgG HCD and has a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the method includes determining that the subject has the presence of IgG HCD and has a serum sBCMA level greater than about 500 ng / mL. In some embodiments, the subject is determined to have the presence of IgG HCD and have a serum sBCMA level greater than about 600 ng / mL. In some embodiments, the subject is determined to have the presence of IgG HCD and have a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the subject is determined to have the presence of IgG HCD and have a serum sBCMA level greater than about 500 ng / mL. In some embodiments, if the subject is determined to have the presence of IgG HCD and have a serum sBCMA level greater than about 600 ng / mL, the method includes debulking the MM. In some embodiments, the method includes debulking the MM when the subject is determined to have the presence of IgG HCD and a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the method includes debulking the MM when the subject is determined to have the presence of IgG HCD and a serum sBCMA level greater than about 500 ng / mL. In some embodiments, the MM is debulked when the subject is determined to have the presence of IgG HCD and a serum sBCMA level greater than about 600 ng / mL. In some embodiments, the MM is debulked when the subject is determined to have the presence of IgG HCD and a serum sBCMA level greater than about 566 ng / mL. In some embodiments, the MM is debulked when the subject is determined to have the presence of IgG HCD and a serum sBCMA level greater than about 500 ng / mL.In some embodiments, a subject is selected for debulking if the subject is determined to have the presence of IgG HCD and a serum sBCMA level greater than about 600 ng / mL. In some embodiments, a subject is selected for debulking if the subject is determined to have the presence of IgG HCD and a serum sBCMA level greater than about 566 ng / mL. In some embodiments, a subject is selected for debulking if the subject is determined to have the presence of IgG HCD and a serum sBCMA level greater than about 500 ng / mL.

[0175] In some embodiments, the method comprises: (a) determining that a subject has (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL and / or (ii) the presence of IgG heavy chain disease (HCD); (b) debulking the MM; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method comprises: (a) determining that a subject has a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method comprises: (a) determining that a subject has the presence of IgG heavy chain disease (HCD); (b) debulking the MM; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method includes: (a) determining that the subject has (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL and (ii) the presence of IgG heavy chain disease (HCD); (b) debulking the MM; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA.

[0176] In some embodiments, the method comprises: (a) determining that a subject has (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 566 ng / mL and / or (ii) the presence of IgG heavy chain disease (HCD); (b) debulking the MM; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method comprises: (a) determining that a subject has a serum soluble B-cell maturation antigen (sBCMA) level greater than about 566 ng / mL; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method comprises: (a) determining that a subject has the presence of IgG heavy chain disease (HCD); (b) debulking the MM; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method includes: (a) determining that the subject has (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 566 ng / mL and (ii) the presence of IgG heavy chain disease (HCD); (b) debulking the MM; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA.

[0177] In some embodiments, the method comprises: (a) determining that a subject has (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 500 ng / mL and / or (ii) the presence of IgG heavy chain disease (HCD); (b) debulking the MM; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method comprises: (a) determining that a subject has a serum soluble B-cell maturation antigen (sBCMA) level greater than about 500 ng / mL; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method comprises: (a) determining that a subject has the presence of IgG heavy chain disease (HCD); (b) debulking the MM; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells that express a chimeric antigen receptor (CAR) that binds BCMA. In some embodiments, the method includes: (a) determining that the subject has (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 500 ng / mL and (ii) the presence of IgG heavy chain disease (HCD); (b) debulking the MM; and (c) administering to the subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA.

[0178] In some embodiments, the method comprises administering to the subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein (a) the subject has (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL; and / or (ii) the presence of IgG heavy chain disease (HCD); and (b) the MM has been debulked between (1) the subject's determination of (i) and / or (ii) and (2) the subject's administration of the T cell therapy. In some embodiments, the method comprises administering to the subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein (a) the subject has previously been determined to have a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL; and (b) the MM has been debulked between (1) the subject's determination of (i) and / or (ii) and (2) the subject's administration of the T cell therapy. In some embodiments, the method comprises administering to a subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein (a) the subject has been previously determined to have the presence of IgG heavy chain disease (HCD); and (b) the MM has been debulked between (1) the time the subject was determined to have (i) and / or (ii) and (2) the time the subject was administered the T cell therapy. In some embodiments, the method comprises administering to a subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein (a) the subject has (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL; and (ii) the time the subject was previously determined to have the presence of IgG heavy chain disease (HCD); and (b) the MM has been debulked between (1) the time the subject was determined to have (i) and / or (ii) and (2) the time the subject was administered the T cell therapy.

[0179] In some embodiments, the method comprises administering to the subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein (a) the subject has (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 566 ng / mL; and / or (ii) the presence of IgG heavy chain disease (HCD); and (b) the MM has been debulked between (1) the subject's determination of (i) and / or (ii) and (2) the subject's administration of the T cell therapy. In some embodiments, the method comprises administering to the subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein (a) the subject has previously been determined to have a serum soluble B-cell maturation antigen (sBCMA) level greater than about 566 ng / mL; and (b) the MM has been debulked between (1) the subject's determination of (i) and / or (ii) and (2) the subject's administration of the T cell therapy. In some embodiments, the method comprises administering to a subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein (a) the subject has been previously determined to have the presence of IgG heavy chain disease (HCD); and (b) the MM has been debulked between (1) the time the subject was determined to have (i) and / or (ii) and (2) the time the subject was administered the T cell therapy. In some embodiments, the method comprises administering to a subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein (a) the subject has (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 566 ng / mL; and (ii) the time the subject was previously determined to have the presence of IgG heavy chain disease (HCD); and (b) the MM has been debulked between (1) the time the subject was determined to have (i) and / or (ii) and (2) the time the subject was administered the T cell therapy.

[0180] In some embodiments, the method includes administering to a subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein (a) the subject is previously determined to have (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 500 ng / mL; and / or (ii) the presence of IgG heavy chain disease (HCD); and (b) the MM has been debulked between (1) the time the subject is determined to have (i) and / or (ii) and (2) the time the subject is administered the T cell therapy. In some embodiments, the method comprises administering to the subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein (a) the subject is predetermined to have a serum soluble B-cell maturation antigen (sBCMA) level greater than about 500 ng / ml; and (b) the MM has been debulked between (1) the subject's determination that they have (i) and / or (ii) and (2) the subject's administration of the T cell therapy. In some embodiments, the method comprises administering to the subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein (a) the subject is predetermined to have the presence of IgG heavy chain disease (HCD); and (b) the MM has been debulked between (1) the subject's determination that they have (i) and / or (ii) and (2) the subject's administration of the T cell therapy. In some embodiments, the method includes administering to a subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds BCMA, wherein (a) the subject is previously determined to have (i) a serum soluble B-cell maturation antigen (sBCMA) level greater than about 500 ng / mL; and (ii) the presence of IgG heavy chain disease (HCD); and (b) the MM has been debulked between (1) the time the subject is determined to have (i) and / or (ii) and (2) the time the subject is administered the T cell therapy.

[0181] In some embodiments, the method includes determining that the subject has a first serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL. In some embodiments, the method includes determining that the subject has a first serum soluble B-cell maturation antigen (sBCMA) level greater than about 566 ng / mL. In some embodiments, the method includes determining that the subject has a first serum soluble B-cell maturation antigen (sBCMA) level greater than about 500 ng / mL. In some embodiments, the subject has been determined to have a first serum sBCMA level greater than about 600 ng / mL. In some embodiments, the subject has been determined to have a first serum sBCMA level greater than about 566 ng / mL. In some embodiments, the subject has been determined to have a first serum sBCMA level greater than about 500 ng / mL. In some embodiments, if the subject is determined to have a first serum sBCMA level greater than about 600 ng / mL, the method includes debulking the MM. In some embodiments, the method includes debulking the MM when the subject is determined to have a first serum sBCMA level greater than about 566 ng / mL. In some embodiments, the method includes debulking the MM when the subject is determined to have a first serum sBCMA level greater than about 500 ng / mL. In some embodiments, the MM is debulked when the subject is determined to have a first serum sBCMA level greater than about 600 ng / mL. In some embodiments, the MM is debulked when the subject is determined to have a first serum sBCMA level greater than about 566 ng / mL. In some embodiments, the MM is debulked when the subject is determined to have a first serum sBCMA level greater than about 500 ng / mL. In some embodiments, the subject is selected for debulking when the subject is determined to have a first serum sBCMA level greater than about 600 ng / mL. In some embodiments, if the subject is determined to have a first serum sBCMA level greater than about 566 ng / mL, the subject is selected for debulking.In some embodiments, if the subject is determined to have a first serum sBCMA level greater than about 500 ng / mL, the subject is selected for debulking.

[0182] In some embodiments, after debulking, the method comprises determining that the subject has a post-debulking serum soluble B-cell maturation antigen (sBCMA) level of less than about 600 ng / mL. In some embodiments, after debulking, the method comprises determining that the subject has a post-debulking serum soluble B-cell maturation antigen (sBCMA) level of less than about 566 ng / mL. In some embodiments, after debulking, the method comprises determining that the subject has a post-debulking serum soluble B-cell maturation antigen (sBCMA) level of less than about 500 ng / mL. In some embodiments, after debulking, the subject is determined to have a post-debulking serum sBCMA level of less than about 600 ng / mL. In some embodiments, after debulking, the subject is determined to have a post-debulking serum sBCMA level of less than about 566 ng / mL. In some embodiments, after debulking, the subject is determined to have a post-debulking serum sBCMA level of less than about 500 ng / mL. In some embodiments, if the subject is determined to have a post-debulking serum sBCMA level below about 600 ng / mL, the method comprises administering T cell therapy to the subject. In some embodiments, if the subject is determined to have a post-debulking serum sBCMA level below about 566 ng / mL, the method comprises administering T cell therapy to the subject. In some embodiments, if the subject is determined to have a post-debulking serum sBCMA level below about 500 ng / mL, the method comprises administering T cell therapy to the subject. In some embodiments, if the subject is determined to have a post-debulking serum sBCMA level below about 600 ng / mL, T cell therapy is administered to the subject. In some embodiments, if the subject is determined to have a post-debulking serum sBCMA level below about 566 ng / mL, T cell therapy is administered to the subject. In some embodiments, if the subject is determined to have a post-debulking serum sBCMA level below about 500 ng / mL, T cell therapy is administered to the subject.

[0183] In some embodiments, the method includes: (a) determining that the subject has a first serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL; (b) debulking the MM; (c) determining that the subject has a post-debulking serum sBCMA level less than about 600 ng / mL; and (d) administering to the subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds to BCMA. In some embodiments, if the subject is determined to have a post-debulking serum sBCMA level greater than about 600 ng / mL, the T cell therapy is not administered to the subject.

[0184] In some embodiments, the method includes: (a) determining that the subject has a first serum soluble B-cell maturation antigen (sBCMA) level greater than about 566 ng / mL; (b) debulking the MM; (c) determining that the subject has a post-debulking serum sBCMA level less than about 566 ng / mL; and (d) administering to the subject a T cell therapy comprising a dose of engineered T cells expressing a chimeric antigen receptor (CAR) that binds to BCMA. In some embodiments, if the subject is determined to have a post-debulking serum sBCMA level greater than about 566 ng / mL, the T cell therapy is not administered to the subject.

[0185] In some embodiments, the method includes: (a) determining that the subject has a first serum soluble B-cell maturation antigen (sBCMA) level greater than about 500 ng / mL; (b) debulking the MM; (c) determining that the subject has a post-debulking serum sBCMA level less than about 500 ng / mL; and (d) administering to the subject a T cell therapy comprising a dose of genetically engineered T cells expressing a chimeric antigen receptor (CAR) that binds to BCMA. In some embodiments, if the subject is determined to have a post-debulking serum sBCMA level greater than about 500 ng / mL, the T cell therapy is not administered to the subject.

[0186] In some embodiments, MM is not debulked if the subject (i) has serum sBCMA less than about 590 ng / mL, about 580 ng / mL, about 570 ng / mL, about 560 ng / mL, about 550 ng / mL, about 540 ng / mL, about 530 ng / mL, about 520 ng / mL, about 510 ng / mL, or about 500 ng / mL; and / or (ii) does not have the presence of IgG HCD. In some embodiments, MM is not debulked if the subject has serum sBCMA less than about 590 ng / mL, about 580 ng / mL, about 570 ng / mL, about 560 ng / mL, about 550 ng / mL, about 540 ng / mL, about 530 ng / mL, about 520 ng / mL, about 510 ng / mL, or about 500 ng / mL. In some embodiments, a subject is not selected for debulking if they (i) have a serum sBCMA level of less than about 590 ng / mL, about 580 ng / mL, about 570 ng / mL, about 560 ng / mL, about 550 ng / mL, about 540 ng / mL, about 530 ng / mL, about 520 ng / mL, about 510 ng / mL, or about 500 ng / mL; and / or (ii) do not have the presence of IgG HCD. In some embodiments, a subject is not selected for debulking if they have a serum sBCMA level of less than about 590 ng / mL, about 580 ng / mL, about 570 ng / mL, about 560 ng / mL, about 550 ng / mL, about 540 ng / mL, about 530 ng / mL, about 520 ng / mL, about 510 ng / mL, or about 500 ng / mL. In some embodiments, if a subject has a serum sBCMA level below about 600 ng / mL, the MM is not debulked prior to administration of T cell therapy to the subject. In some embodiments, if a subject has a serum sBCMA level below about 600 ng / mL, the subject is not selected for debulking prior to administration of T cell therapy to the subject. In some embodiments, if a subject has a serum sBCMA level below about 566 ng / mL, the MM is not debulked prior to administration of T cell therapy to the subject. In some embodiments, if a subject has a serum sBCMA level below about 566 ng / mL, the subject is not selected for debulking prior to administration of T cell therapy to the subject.

[0187] In some embodiments, debulking comprises administering chemotherapy, radiation, or an immunomodulatory agent to the subject. In some embodiments, debulking comprises administering chemotherapy to the subject. In some embodiments, the chemotherapy comprises melphalan, doxorubicin, or cyclophosphamide chemotherapy. In some embodiments, debulking comprises administering radiation to the subject. In some embodiments, the immunomodulatory agent is thalidomide, lenalidomide, or pomalidomide. In some embodiments, it comprises administering an immunomodulatory agent to the subject. In some embodiments, the immunomodulatory agent is a checkpoint inhibitor.

[0188] In some of any of the provided embodiments, debulking is performed prior to administration of the T cell therapy to the subject.

[0189] In some embodiments, debulking the MM is performed within about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week prior to administering T cell therapy to the subject. In some embodiments, debulking the MM is performed within about 3 months prior to administering T cell therapy to the subject. In some embodiments, debulking the MM is performed within about 2 months prior to administering T cell therapy to the subject. In some embodiments, debulking the MM is performed within about 1 month prior to administering T cell therapy to the subject. In some embodiments, debulking the MM is performed within about 3 weeks prior to administering T cell therapy to the subject. In some embodiments, debulking the MM is performed within about 2 weeks prior to administering T cell therapy to the subject. In some embodiments, debulking the MM is performed within about 1 week prior to administering T cell therapy to the subject.

[0190] In some embodiments, debulking the MM is performed about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week prior to administering T cell therapy to the subject. In some embodiments, debulking the MM is performed about 3 months prior to administering T cell therapy to the subject. In some embodiments, debulking the MM is performed about 2 months prior to administering T cell therapy to the subject. In some embodiments, debulking the MM is performed about 1 month prior to administering T cell therapy to the subject. In some embodiments, debulking the MM is performed about 3 weeks prior to administering T cell therapy to the subject. In some embodiments, debulking the MM is performed about 2 weeks prior to administering T cell therapy to the subject. In some embodiments, debulking the MM is performed about 1 week prior to administering T cell therapy to the subject. In some embodiments, debulking is performed before administering lymphodepleting therapy to the subject. In some embodiments, debulking is performed after administering lymphodepleting therapy to the subject. In some embodiments, the subject is treated with a gamma secretase inhibitor prior to administration of the T cell therapy to the subject.

[0191] CT cell therapy dosage and administration In some embodiments of the methods, compositions, combinations, kits and uses provided herein, the treatment comprises administering a T cell therapy (e.g., a CAR-expressing T cell) to the subject. For example, the T cell therapy is an anti-BCMA CAR T cell therapy.

[0192] In some embodiments, the cells used in or administered in connection with the provided methods contain or are engineered to contain engineered receptors, such as engineered antigen receptors, for example, chimeric antigen receptors (CARs) or T cell receptors (TCRs). Among the compositions, pharmaceutical compositions and formulations for administration, such as for adoptive cell therapy, are particularly mentioned. Also provided are therapeutic methods for administering cells and compositions to subjects, for example, patients, in accordance with the provided methods and / or using the provided products or compositions.

[0193] In some embodiments, the cell-based therapy is or includes the administration of cells, e.g., immune cells, e.g., T cells or NK cells, that target molecules expressed on the surface of a lesion, such as a tumor or cancer. In some embodiments, the cells express a recombinant receptor, e.g., a CAR, containing an extracellular ligand-binding domain that specifically binds to an antigen. In some embodiments, the recombinant receptor is a CAR containing an extracellular antigen-recognition domain that specifically binds to BCMA. In some embodiments, the immune cells express a recombinant receptor, such as a chimeric antigen receptor (CAR). In some embodiments, the T cell therapy includes administering T cells engineered to express a chimeric antigen receptor (CAR). In certain embodiments, the cell therapy, e.g., anti-BCMA CAR T cell therapy, is for treating multiple myeloma, e.g., relapsed and refractory (R / R multiple myeloma). In some embodiments, the cells are autologous to the subject. In some embodiments, the cells are allogeneic to the subject. Exemplary engineered cells for administration as cell therapy in the provided methods are described in Section IV.

[0194] The method for administering cells for adoptive cell therapy is known, and can be used in connection with the provided methods, compositions, and products and kits.For example, the method of adoptive T cell therapy is described, for example, in U.S. Patent Application Publication No. 2003 / 0170238 to Gruenberg et al.; U.S. Patent No. 4,690,915 to Rosenberg; Rosenberg (2011) Nat Rev Clin Oncol. 8(10):577-85).For example, see Themeli et al. (2013) Nat Biotechnol. 31(10):928-933; Tsukahara et al. (2013) Biochem Biophys Res Commun 438(1):84-9; Davila et al. (2013) PLoS ONE 8(4):e61338.

[0195] In some embodiments, cell therapy, e.g., adoptive T cell therapy, is performed by autologous transfer, whereby cells are isolated and / or otherwise prepared from a subject to receive cell therapy or from a sample derived from such a subject. Thus, in some aspects, the cells are derived from a subject, e.g., a patient, in need of treatment, and the cells, after isolation and processing, are administered to the same subject.

[0196] In some embodiments, cell therapy, e.g., adoptive T cell therapy, is carried out by allogeneic transfer, whereby cells are isolated and / or otherwise prepared from a subject other than the subject that is to receive or will ultimately receive cell therapy, e.g., a first subject. In such embodiments, the cells are then administered to a different subject of the same species, e.g., a second subject. In some embodiments, the first and second subjects are genetically identical. In some embodiments, the first and second subjects are genetically similar. In some embodiments, the second subject expresses the same HLA class or supertype as the first subject.

[0197] The cells of the T cell therapy can be administered in a composition formulated for administration, or alternatively, in two or more compositions (e.g., two compositions) formulated for separate administration. The dose(s) of cells can include a specific or relative number of cells or engineered cells, and / or two or more subtypes in a defined ratio or composition within the composition, such as CD4+ vs. CD8+ T cells.

[0198] The cells can be administered by any suitable means, for example, by bolus injection, injection, e.g., intravenous or subcutaneous injection, intraocular injection, periocular injection, subretinal injection, intravitreal injection, transseptal injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjectval injection, subconjunctival injection, sub-Tenon injection, retrobulbar injection, peribulbar injection, or posterior juxtascleral delivery. In some embodiments, the cells are administered parenterally, intrapulmonary, intranasally, and, if desired for localized treatment, intralesionally. Parenteral injections include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In some embodiments, a given dose is administered by a single bolus of cells. In some embodiments, a given dose is administered by multiple boluses of cells over a period of, for example, three days or less, or by continuous infusion of cells. In some embodiments, administration of the cell dose or any additional therapy, eg, lymphodepleting therapy, interventional therapy, and / or combination therapy, is performed by exogenous delivery.

[0199] For treatment of disease, the appropriate dosage will depend on the type of disease being treated, the type of cells or recombinant receptor, the severity and course of the disease, previous therapy, the subject's clinical history and response to the cells, and the discretion of the treating physician. The compositions and cells, in some embodiments, are suitably administered to the subject at one time or over a series of treatments.

[0200] In certain embodiments, the cells, or individual populations of cell subtypes, are in the range of about 1 million to about 100 billion cells, and / or that amount of cells per kilogram of body weight, e.g., 1 million to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or a range defined by any two of the foregoing values), e.g., about 10 million to about 100 billion cells (e.g., about 20 million cells, about 30 million cells, about 40 million cells, about 60 million cells, about 70 million cells, about 80 million cells, or a range defined by any two of the foregoing values). 0 million cells, about 90 million cells, about 10 billion cells, about 25 billion cells, about 50 billion cells, about 75 billion cells, about 90 billion cells, or a range defined by any two of the foregoing values), and in some cases, about 100 million cells to about 50 billion cells (e.g., about 120 million cells, about 250 million cells, about 350 million cells, about 450 million cells, about 650 million cells, about 800 million cells, about 900 million cells, about 3 billion cells, about 30 billion cells, about 45 billion cells), or any value therebetween, and / or any value per kilogram of body weight, are administered to a subject. Dosages may vary depending on the specific characteristics of the disease or disorder and / or the patient and / or other treatments.

[0201] In some embodiments, for example, when the subject is a human, the dose is about 1 x 10 8 Fewer than 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs), e.g., about 1 x 10 6 ~1×10 8 In the range of 2 x 10 such cells, for example 6 , 5×10 6 , 1×10 7 , 5×10 7 Or 1 x 10 8 In some embodiments, the dose comprises about 5 x 10 such total cells, or a range between any two of the foregoing values. 8Fewer than 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells or peripheral blood mononuclear cells (PBMCs), e.g., about 1 x 10 8 ~5×10 8 In the range of such cells, e.g., 1.5 x 10 8 , 3×10 8 , or 4.5 × 10 8 such total cells, or a range between any two of the foregoing values.

[0202] The cells can be administered by any suitable means. The cells are administered in a dosage regimen to achieve a therapeutic effect, such as reducing tumor burden. Dosage and administration can depend in part on the debulking administration schedule that is carried out before the start of administration of T cell therapy. Various dosage schedules for T cell therapy include, but are not limited to, single or multiple administrations over various time points, bolus administration, and pulse infusion.

[0203] Preconditioning a subject with immunodepleting (eg, lymphodepleting) therapy can, in some aspects, improve the efficacy of adoptive cellular therapy (ACT).

[0204] Thus, in some embodiments, the method includes administering a preconditioning agent, such as a lymphodepleting or chemotherapeutic agent, such as cyclophosphamide, fludarabine, or a combination thereof, to the subject prior to the initiation of cell therapy. For example, the subject can be administered the preconditioning agent at least 2 days, e.g., at least 3, 4, 5, 6, or 7 days, prior to the initiation of cell therapy. In some embodiments, the subject is administered the preconditioning agent 7 days or less, e.g., 6, 5, 4, 3, or 2 days or less, prior to the initiation of cell therapy.

[0205] In some embodiments, the subject is administered a preconditioning agent (lymphocyte depletion treatment) as described in Section II.D.

[0206] After administration of the cells, the biological activity of the engineered cell population is measured, in some embodiments, by any of a number of known methods, for example. Parameters for evaluation include specific binding of engineered or natural T cells or other immune cells to antigens in vivo, for example, by imaging, or ex vivo, for example, by ELISA or flow cytometry. In certain embodiments, the ability of the engineered cells to destroy target cells can be measured using any suitable known method, such as the cytotoxicity assays described in Kochenderfer et al., J. Immunotherapy, 32(7): 689-702 (2009) and Herman et al. J. Immunological Methods, 285(1): 25-40 (2004). In certain embodiments, the biological activity of the cells is measured by assaying the expression and / or secretion of one or more cytokines, such as CD107a, IFNγ, IL-2, and TNF. In some aspects, biological activity is measured by assessing clinical outcomes, such as reduction in tumor burden or burden.

[0207] In some embodiments, the dose of cells is administered to the subject in accordance with the method of T cell therapy provided.In some embodiments, the size or timing of the dose is determined as a function of the specific disease or condition of the subject.In consideration of the explanation provided, the size or timing of the dose for specific disease can be empirically determined.

[0208] In some embodiments, the cells, or individual populations of cell subtypes, range from about 100,000 to about 100 billion cells, and / or that amount of cells per kilogram of subject body weight, e.g., 100,000 to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or is a range defined by any two of the aforementioned values), for example, 10 million to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or a range defined by any two of the aforementioned values), for example, about 10 million to about 100 billion cells (e.g., about 20 million cells, about 30 million cells, about 40 million cells, about 60 million cells, about 70 million cells, about 80 million cells, about 90 million cells, about 10 billion cells, about 25 billion cells, about 50 billion cells, about 75 billion cells, about 90 billion cells, or a range defined by any two of the foregoing values), and in some cases, about 100 million cells to about 50 billion cells The subject is administered about 120 million cells, about 150 million cells, about 250 million cells, about 350 million cells, about 450 million cells, about 650 million cells, about 800 million cells, about 900 million cells, about 3 billion cells, about 30 billion cells, about 45 billion cells) or any value between these ranges, and / or any value per kilogram of body weight. Dosages may vary depending on the specific characteristics of the disease or disorder and / or patient and / or other treatment. In some embodiments, such values ​​refer to the number of recombinant receptor-expressing cells; in other embodiments, it refers to the number of T cells or PBMCs or total cells administered.

[0209] In some embodiments, the cell therapy comprises at least or at least about 0.1 x 10 6 Cells / kg (subject weight), 0.2 x 110 6 cells / kg, 0.3×10 6cells / kg, 0.4×10 6 cells / kg, 0.5×10 6 cells / kg, 1×10 6 cells / kg, 2.0×10 6 cells / kg, 3×10 6 cells / kg or 5 x 10 6 It includes administering a dose containing a number of cells that is or is about cells / kg.

[0210] In some embodiments, the cell therapy comprises 0.1 x 10 6 Cells / kg (subject weight) ~ 1.0 x 10 7 Between or around 0.5 x 10 cells / kg 6 cells / kg~5×10 6 Between or around 0.5 x 10 cells / kg 6 cells / kg~3×10 6 Between or around 0.5 x 10 cells / kg 6 cells / kg~2×10 6 Between or around 0.5 x 10 cells / kg 6 cells / kg~1×10 6 Between or about 1.0 x 10 cells / kg 6 ~5 x 10 cells / kg (subject weight) 6 Between or about 1.0 x 10 cells / kg 6 cells / kg~3×10 6 Between or about 1.0 x 10 cells / kg 6 cells / kg~2×10 6 Between or about 2.0 x 10 cells / kg 6 ~5 x 10 cells / kg (subject weight) 6 Between or about 2.0 x 10 cells / kg 6 cells / kg~3×10 6 cells / kg or 3.0 x 10 6 ~5 x 10 cells / kg (subject weight) 6 This includes administering a dose containing a number of cells between or about (inclusive) cells / kg.

[0211] In some embodiments, the dose of cells is 2×10 5 or approximate number of cells / kg ~ 2 x 10 6 or approximately 4 x 10 cells / kg 5 or approximately 1 x 10 cells / kg 6 Between 6 x 10 or the approximate number of cells / kg 5 or approximately 8 x 10 cells / kg 5 In some embodiments, the dose of cells is between 2 x 10 cells / kg or about 2 x 10 cells / kg of subject body weight. 5 3×10 or less cells (e.g., antigen-expressing, e.g., CAR-expressing cells) (cells / kg), e.g., 3×10 5 Cells / kg or less, 4 x 10 5 Cells / kg or less, 5 x 10 5 Cells / kg or less, 6 x 10 5 7 x 10 cells / kg or less 5 Cells / kg or less, 8 x 10 5 Cells / kg or less, 9 x 10 5 Cells / kg or less, 1 x 10 6 Cells / kg or less, or 2 x 10 6 In some embodiments, the dose of cells comprises at least about 2 x 10 cells / kg or less. 5 or at or about that number of cells (e.g., antigen-expressing, e.g., CAR-expressing cells) (cells / kg), e.g., at least or at least about 3 x 10 5 cells / kg or an approximation thereof, at least or at least about 4 x 10 5 cells / kg or an approximation thereof, at least or at least about 5 x 10 5 cells / kg or an approximation thereof, at least or at least about 6 x 10 5 cells / kg or an approximation thereof, at least or at least about 7 x 10 5cells / kg or an approximation thereof, at least or at least about 8 x 10 5 cells / kg or an approximation thereof, at least or at least about 9 x 10 5 cells / kg or an approximation thereof, at least or at least about 1 x 10 6 cells / kg or at or about that number, or at least about 2 x 10 6 Includes cells / kg or any value or approximation thereof.

[0212] In some embodiments, the dose of cells is a flat dose of cells or a fixed dose of cells such that the dose of cells is not related to or based on the body surface area or weight of the subject.

[0213] In some embodiments, the cell therapy comprises 1 x 10 5 ~2×10 9 5 x 10 or approximately 5 x 10 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs) 5 ~1×10 9 or approximately 1 x 10 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), or 1 x 10 6 ~1×10 9 In some embodiments, the cell therapy comprises administering a dose comprising at or about 1 x 10 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), inclusive. 5 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), for example, at least or about at least 1 x 10 6 , at least or about at least 1 x 10 7 , at least or about at least 1 x 10 8 , at least or about at least 1 x 10 9 It includes administering a dose of cells, including a number of such cells.

[0214] In some embodiments, the dose of genetically engineered cells is at least or at least about 1 x 10 5 CAR-expressing cells, at least or at least about 2.5 x 10 5 CAR-expressing cells, at least or at least about 5 x 10 5 CAR-expressing cells, at least or at least about 1 x 10 6 6 CAR-expressing cells, at least or at least about 2.5 x 10 6 CAR-expressing cells, at least or at least about 5 x 10 6 CAR-expressing cells, at least or at least about 1 x 10 7 CAR-expressing cells, at least or at least about 2.5 x 10 7 CAR-expressing cells, at least or at least about 5 x 10 7 CAR-expressing cells, at least or at least about 1 x 10 8 CAR-expressing cells, at least or at least about 2.5 x 10 8 CAR-expressing cells, or at least about or at least 5 x 10 8 Contains CAR-expressing cells.

[0215] In some embodiments, for example, when the subject is a human, the dose is 1×10 6 or greater than approximately 2 x 10 total recombinant receptor (e.g., CAR)-expressing (CAR+) cells, T cells, or peripheral blood mononuclear cells (PBMCs) 9 or less than the approximate number of total recombinant receptor (e.g., CAR)-expressing cells, T cells or peripheral blood mononuclear cells (PBMCs), e.g., 1.0 x 10 7 Pieces or their approximate number ~ 1.2 x 10 9 or in the range of approximately 1.0 x 10 such cells, e.g., 7 , 1.5×10 7 , 2.0×10 7 , 2.5×10 7 , 5×10 7 , 1.5×10 8 , 3×10 8 , 4.5×10 8 , 6×108 , 8×10 8 or 1.2 x 10 9 In some embodiments, for example, when the subject is a human, the dose is 1 x 10 or about 1 x 10 such total cells. 6 or greater than approximately 2 x 10 total recombinant receptor (e.g., CAR)-expressing (CAR+) cells, T cells, or peripheral blood mononuclear cells (PBMCs) 9 or less than the approximate number of total recombinant receptor (e.g., CAR)-expressing cells, T cells or peripheral blood mononuclear cells (PBMCs), e.g., 2.5 x 10 7 Pieces or their approximate number ~ 1.2 x 10 9 or in the range of approximately 2.5 x 10 such cells 7 , 5×10 7 , 1.5×10 8 , 3×10 8 , 4.5×10 8 , 6×10 8 , 8×10 8 or 1.2 x 10 9 In some embodiments, for example, when the subject is a human, the dose is 1.0 x 10 or about 1.0 x 10 such total cells. 7 In some embodiments, for example, when the subject is a human, the dose comprises 1.5 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 7 In some embodiments, for example, when the subject is a human, the dose comprises 2.0 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 7 In some embodiments, for example, when the subject is a human, the dose comprises 2.5 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 7 In some embodiments, for example, when the subject is a human, the dose comprises 5×10 or approximately 5×10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 7In some embodiments, for example, when the subject is a human, the dose comprises 1.5 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 8 In some embodiments, for example, when the subject is a human, the dose comprises 3 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 8 In some embodiments, for example, when the subject is a human, the dose comprises 4.5 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 8 In some embodiments, for example, when the subject is a human, the dose comprises 6 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 8 In some embodiments, for example, if the subject is a human, the dose comprises 8 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 8 In some embodiments, for example, when the subject is a human, the dose comprises 1.2 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). 9 The cells comprise at or about the total number of recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs).

[0216] In some embodiments, the dose of genetically engineered cells is 1×10 5 pieces or the approximate number ~ 2 x 10 9 Total CAR-expressing (CAR+) T cells, 1 x 10 5 pieces or the approximate number ~ 5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or its approximate number ~ 2.5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or its approximate number ~ 1 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 5pieces or the approximate number ~ 5 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or its approximate number ~ 2.5 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or its approximate number ~ 1 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or the approximate number ~ 5 x 10 6 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or its approximate number ~ 2.5 x 10 6 1 x 10 total CAR-expressing T cells or their approximate number 5 pieces or its approximate number ~ 1 x 10 6 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or the approximate number ~ 5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 2.5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 1 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or the approximate number ~ 5 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 2.5 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 1 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or the approximate number ~ 5 x 10 6 1 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 2.5 x 10 6 Total CAR-expressing T cells, 2.5 x 10 6 pieces or the approximate number ~ 5 x 10 8 Total CAR-expressing T cells, 2.5 x 10 6 pieces or its approximate number ~ 2.5 x 10 8Total CAR-expressing T cells, 2.5 x 10 6 pieces or its approximate number ~ 1 x 10 8 Total CAR-expressing T cells, 2.5 x 10 6 pieces or the approximate number ~ 5 x 10 7 Total CAR-expressing T cells, 2.5 x 10 6 pieces or its approximate number ~ 2.5 x 10 7 Total CAR-expressing T cells, 2.5 x 10 6 pieces or its approximate number ~ 1 x 10 7 Total CAR-expressing T cells, 2.5 x 10 6 pieces or the approximate number ~ 5 x 10 6 5 x 10 total CAR-expressing T cells or their approximate number 6 pieces or the approximate number ~ 5 x 10 8 5 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 2.5 x 10 8 5 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 1 x 10 8 5 x 10 total CAR-expressing T cells or their approximate number 6 pieces or the approximate number ~ 5 x 10 7 5 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 2.5 x 10 7 5 x 10 total CAR-expressing T cells or their approximate number 6 pieces or its approximate number ~ 1 x 10 7 1 x 10 total CAR-expressing T cells or their approximate number 7 pieces or the approximate number ~ 5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 7 pieces or its approximate number ~ 2.5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 7 pieces or its approximate number ~ 1 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 7 pieces or the approximate number ~ 5 x 10 71 x 10 total CAR-expressing T cells or their approximate number 7 pieces or its approximate number ~ 2.5 x 10 7 Total CAR-expressing T cells, 2.5 x 10 7 pieces or the approximate number ~ 5 x 10 8 Total CAR-expressing T cells, 2.5 x 10 7 pieces or its approximate number ~ 2.5 x 10 8 Total CAR-expressing T cells, 2.5 x 10 7 pieces or its approximate number ~ 1 x 10 8 Total CAR-expressing T cells, 2.5 x 10 7 pieces or the approximate number ~ 5 x 10 7 5 x 10 total CAR-expressing T cells or their approximate number 7 pieces or the approximate number ~ 5 x 10 8 5 x 10 total CAR-expressing T cells or their approximate number 7 pieces or its approximate number ~ 2.5 x 10 8 5 x 10 total CAR-expressing T cells or their approximate number 7 pieces or its approximate number ~ 1 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 8 pieces or the approximate number ~ 5 x 10 8 1 x 10 total CAR-expressing T cells or their approximate number 8 pieces or its approximate number ~ 2.5 x 10 8 or approximately 2.5 x 10 total CAR-expressing T cells 8 pieces or the approximate number ~ 5 x 10 8 In some embodiments, the dose of engineered cells comprises at or about 1.0 x 10 total CAR-expressing T cells. 7 pieces or the approximate number ~ 8 x 10 8 Total CAR-expressing (CAR+) T cells, 1.0 x 10 7 pieces or its approximate number ~ 6.5 x 10 8 Total CAR+ T cells, 1.5 x 10 7 pieces or its approximate number ~ 6.5 x 10 8 Total CAR+ T cells, 1.5 x 10 7pieces or its approximate number ~ 6.0 x 10 8 Total CAR+ T cells, 2.5 x 10 7 pieces or its approximate number ~ 6.0 x 10 8 or approximately 5.0 x 10 total CAR+ T cells 7 pieces or its approximate number ~ 6.0 x 10 8 Contains at or about the total number of CAR+ T cells.

[0217] In some embodiments, the dose of genetically engineered cells is 2.5×10 7 Approximately 1.2 x 10 or approximately 1.2 x 10 CAR-expressing (CAR+) T cells, total T cells, or total peripheral blood mononuclear cells (PBMCs) 9 Between 5.0 x 10 or the approximate number of CAR-expressing T cells, total T cells, or total PBMCs 7 or approximate number of CAR-expressing T cells, total T cells, or total peripheral blood mononuclear cells (PBMCs) ~6.0 x 10 8 Between 5.0 x 10 or the approximate number of CAR-expressing T cells, total T cells, or total PBMCs 7 or approximate number of CAR-expressing T cells ~4.5 x 10 8 Between 1.5 x 10 or the approximate number of CAR-expressing T cells, total T cells, or total peripheral blood mononuclear cells (PBMCs) 8 or approximate number of CAR-expressing T cells ~3.0 x 10 8 In some embodiments, the dose comprises between 2.5 x 10 and approximately 2.5 x 10 CAR-expressing T cells, total T cells, or total PBMCs, inclusive. In some embodiments, the number refers to the total number of CD3+ or CD8+, and in some cases, CAR-expressing (e.g., CAR+) cells. In some embodiments, the dose is between 2.5 x 10 and approximately 2.5 x 10 7 Pieces or their approximate number ~ 1.2 x 10 9 5.0 x 10 or the approximate number of CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells 7 Pieces or their approximate number ~ 6.0 x 10 8 5.0 x 10 or the approximate number of CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells 7 Pieces or their approximate number ~ 4.5 x 10 8or approximately 1.5 x 10 CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells 8 Pieces or their approximate number ~ 3.0 x 10 8 The number of cells includes the number of CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells (inclusive) at or near that number.

[0218] In some embodiments, the dose is 1.0 x 10 7 In some embodiments, the dose is 1.5 x 10 CD3+ CAR-expressing cells. 7 In some embodiments, the dose is 2.0 x 10 CD3+ CAR-expressing cells. 7 In some embodiments, the dose is 2.5 x 10 CD3+ CAR-expressing cells. 7 In some embodiments, the dose is 5 x 10 CD3+ CAR-expressing cells. 7 In some embodiments, the dose is 1.5 x 10 CD3+ CAR-expressing cells. 8 In some embodiments, the dose is 3 x 10 CD3+ CAR-expressing cells. 8 In some embodiments, the dose is 4.5 x 10 CD3+ CAR-expressing cells. 8 In some embodiments, the dose is 6 x 10 CD3+ CAR-expressing cells. 8 In some embodiments, the dose is 8 x 10 CD3+ CAR-expressing cells. 8 In some embodiments, the dose is 1.2 x 10 CD3+ CAR-expressing cells. 9 CD3+CAR-expressing cells.

[0219] In some embodiments, the dose of engineered cells refers to the total number of CD3+ CAR-expressing (CAR+) or CD4+ / CD8+ CAR-expressing (CAR+) cells. In some embodiments, the dose is 1.0 x 10 7 Pieces or their approximate number ~ 1.2 x 10 9 1.5 x 10 or the approximate number of CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells 7Pieces or their approximate number ~ 1.2 x 10 9 2.0 x 10 or approximate number of CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells 7 Pieces or their approximate number ~ 1.2 x 10 9 2.5 x 10 or the approximate number of CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells 7 Pieces or their approximate number ~ 1.2 x 10 9 5.0 x 10 or the approximate number of CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells 7 Pieces or their approximate number ~ 6.0 x 10 8 5.0 x 10 or the approximate number of CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells 7 Pieces or their approximate number ~ 4.5 x 10 8 or approximately 1.5 x 10 CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells 8 Pieces or their approximate number ~ 3.0 x 10 8 In some embodiments, the dose comprises 1.0 x 10 or approximately 1.0 x 10 total CD3+ or CD4+ / CD8+ T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells (inclusive) of engineered cells. 7 , 1.5×10 7 , 2.0×10 7 , 2.5×10 7 , 5×10 7 , 1.5×10 8 , 3×10 8 , 4.5×10 8 , 6×10 8 , 8×10 8 or 1.2 x 10 9 In some embodiments, the dose comprises 2.5 x 10 total CD3+ or CD4+ / CD8+ T cells or CD3+ CAR-expressing or CD4+ / CD8+ CAR-expressing cells. 7, 5×10 7 , 1.5×10 8 , 3×10 8 , 4.5×10 8 , 6×10 8 , 8×10 8 or 1.2 x 10 9 In some embodiments, the dose comprises 1.0 x 10 CD3+ CAR-expressing cells or approximately 1.0 x 10 7 , 1.5×10 7 , 2.0×10 7 , 2.5×10 7 , 5×10 7 , 1.5×10 8 , 3×10 8 , 4.5×10 8 , 6×10 8 , 8×10 8 or 1.2 x 10 9 The cells contain at or about that number of CD4+ / CD8+ CAR-expressing cells.

[0220] In some embodiments, the dose is 1.0 x 10 7 In some embodiments, the dose is 1.5 x 10 7 In some embodiments, the dose is 2.0 x 10 7 In some embodiments, the dose is 2.5 x 10 CD4+ / CD8+ CAR-expressing cells or approximately 2.5 x 10 7 In some embodiments, the dose is 5×10 7 In some embodiments, the dose is 1.5 x 10 8 In some embodiments, the dose is 3 x 10 8 In some embodiments, the dose is 4.5 x 10 CD4+ / CD8+ CAR-expressing cells or approximately 4.5 x 10 8 In some embodiments, the dose is 6 x 10 CD4+ / CD8+ CAR-expressing cells or approximately 6 x 10 8In some embodiments, the dose is 8 x 10 CD4+ / CD8+ CAR-expressing cells or approximately 8 x 10 8 In some embodiments, the dose is 1.2 x 10 9 In some embodiments, the dose is 2.5 x 10 CD4+ / CD8+ CAR-expressing cells or approximately 2.5 x 10 7 In some embodiments, the dose is 5 x 10 7 In some embodiments, the dose is 1.5 x 10 8 In some embodiments, the dose is 3 x 10 8 In some embodiments, the dose is 4.5 x 10 8 In some embodiments, the dose is 6 x 10 8 In some embodiments, the dose is 6.5 x 10 8 In some embodiments, the dose is 8 x 10 8 In some embodiments, the dose is 1.2 x 10 9 or approximately that number of CD4+ or CD8+ CAR-expressing cells.

[0221] In some embodiments, the dose of T cells comprises CD4+ T cells, CD8+ T cells, or CD4+ and CD8+ T cells. In some embodiments, the dose of T cells comprises CD4+ T cells. In some embodiments, the dose of T cells comprises CD8+ T cells. In some embodiments, the dose of T cells comprises CD4+ T cells or CD8+ T cells. In some embodiments, the dose of T cells comprises CD4+ and CD8+ T cells.

[0222] In some embodiments, for example, when the subject is a human, the dose of CD4+ T cells and CD8+ T cells totals 1×10 6 Pieces or their approximate number ~ 2 x 10 9 Between or about 2.5 x 10 total CAR-expressing CD4+ cells and CAR-expressing CD8+ cells, for example, 2.5 x 10 7 Pieces or their approximate number ~ 1.2 x 10 9 or in the range of approximately 5 x 10 such cells 7 Pieces or their approximate number ~ 4.5 x 10 8 in the range of or about that number of such cells; e.g., 1.0 x 10 7 Pieces or its approximate number, 2.5 x 10 7 Pieces or its approximate number, 2.0 x 10 7 Pieces or its approximate number, 2.5 x 10 7 Pieces or its approximate number, 5 x 10 7 Pieces or its approximate number, 1.5 x 10 8 Pieces or its approximate number, 3 x 10 8 Pieces or its approximate number, 4.5 x 10 8 Pieces or its approximate number, 6 x 10 8 pieces or its approximate number, 6.5 x 10 8 Pieces or its approximate number, 8 x 10 8 or its approximate number, or 1.2 x 10 9 In some embodiments, for example, when the subject is a human, the dose of CD8+ T cells, including in the dose comprising CD4+ T cells and CD8+ T cells, is 1 x 10 or about 1 x 10 such total cells, or a range between any two of the foregoing values. 6 Pieces or their approximate number ~ 2 x 10 9 Between or about 10 total recombinant receptor (e.g., CAR)-expressing CD8+ cells, e.g., 2.5 x 10 7 Pieces or their approximate number ~ 1.2 x 10 9 or in the range of approximately 5 x 10 such cells 7 Pieces or their approximate number ~ 4.5 x 10 8 in the range of or about that number of such cells; e.g., 2.5 x 10 7 Pieces or its approximate number, 5 x 10 7Pieces or its approximate number, 1.5 x 10 8 Pieces or its approximate number, 3 x 10 8 Pieces or its approximate number, 4.5 x 10 8 Pieces or its approximate number, 6 x 10 8 Pieces or its approximate number, 8 x 10 8 or its approximate number, or 1.2 x 10 9 This includes at or about that number of such total cells, or a range between any two of the foregoing values.

[0223] In some embodiments, a dose of cells, e.g., recombinant receptor-expressing T cells, is administered to a subject as a single dose, or is administered only once within a period of two weeks, one month, three months, six months, one year, or more. In some embodiments, a patient is administered multiple doses, and each or the total dose can be within any of the values ​​set forth above. In some embodiments, the engineered cells or compositions of engineered cells for administration exhibit characteristics indicative of or consistent with cell health. In some embodiments, 70, 75, 80, 85, or 90% of such doses, or at least about 70, 75, 80, 85, or 90% of the CAR+ cells exhibit one or more characteristics or phenotypes indicative of cell health or biologically active CAR cells, such as the absence of expression of apoptotic markers.

[0224] In certain embodiments, the phenotype is or includes the absence of apoptosis and / or the absence of indicators that the cell is undergoing the apoptotic process. Apoptosis is a process of programmed cell death that involves a series of stereotypical morphological and biochemical events that result in characteristic cellular changes and death, including blebbing, cell shrinkage, nuclear fragmentation, chromatin condensation, chromosomal DNA fragmentation, and general mRNA decay. In some aspects, the early stage of apoptosis can be indicated by the activation of certain caspases, such as 2, 8, 9, and 10. In some aspects, the mid- to late-stage of apoptosis is characterized by further loss of membrane integrity, chromatin condensation, and DNA fragmentation, and includes biochemical events such as the activation of caspases 3, 6, and 7.

[0225] In certain embodiments, the phenotype is one or more factors associated with programmed cell death, e.g., pro-apoptotic factors known to initiate apoptosis, e.g., members of the death receptor pathway, activated members of the mitochondrial (intrinsic) pathway, e.g., Bcl-2 family members, e.g., Bax, Bad, and Bid, and negative expression of caspases. In certain embodiments, the phenotype is an indicator, e.g., the absence of annexin V molecules or TUNEL staining, which will preferentially bind to cells undergoing apoptosis when incubated or contacted with the cell composition. In some embodiments, the phenotype is or includes expression of one or more markers indicative of an apoptotic state in cells. In some embodiments, the phenotype is the lack of expression and / or activation of a caspase, e.g., caspase-3. In some aspects, activation of caspase-3 indicates increased or resurrected apoptosis. In certain embodiments, caspase activation can be detected by known methods. In some embodiments, caspase activation can be detected using an antibody that specifically binds to activated caspase (i.e., specifically binds to the cleaved polypeptide). In certain embodiments, the phenotype is or includes active caspase 3. In some embodiments, the marker of apoptosis is a reagent that detects a feature in cells associated with apoptosis. In certain embodiments, the reagent is an annexin V molecule.

[0226] In some embodiments, compositions containing engineered cells for administration contain a certain number or amount of cells that exhibit a phenotype indicative of or consistent with cellular health. In some of any of the embodiments, less than about 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the CAR-expressing T cells in a dose of engineered T cells express a marker of apoptosis, optionally Annexin V or active Caspase 3. In some of any of the embodiments, less than 5%, 4%, 3%, 2%, or 1% of the CAR-expressing T cells in a dose of engineered T cells express Annexin V or active Caspase 3.

[0227] In the context of adoptive cell therapy, administration of a given "dose" of cells encompasses administration of a given amount or number of cells as a single composition and / or a single uninterrupted administration, e.g., as a single injection or continuous infusion, as well as administration of a given amount or number of cells as divided doses provided in multiple individual compositions or infusions over a specified period of time, which is three days or less. Thus, in some situations, a dose is a single or continuous administration of a specified number of cells given or initiated at a single time point. However, in some situations, a dose is administered in multiple injections or infusions over a period of three days or less, such as once daily for three or two days, or by multiple infusions over a one-day period.

[0228] Thus, in some aspects, the dose of cells is administered in a single pharmaceutical composition, hi some embodiments, the dose of cells is administered in multiple compositions that collectively contain the dose of cells.

[0229] The term "split dose" refers to a dose that is divided so as to be administered over more than one day. This type of administration is encompassed by the present method and is considered to be a single dose. In some embodiments, the split dose cells are administered in multiple compositions that collectively comprise the dose cells over a period of 3 days or less.

[0230] Thus, the dose of cells can be administered as a split dose. For example, in some embodiments, the dose can be administered to a subject over two or three days. An exemplary method for split dosing includes administering 25% of the dose on day 1 and the remaining 75% of the dose on day 2. In other embodiments, 33% of the dose can be administered on day 1 and the remaining 67% on day 2. In some aspects, 10% of the dose is administered on day 1, 30% of the dose is administered on day 2, and 60% of the dose is administered on day 3. In some embodiments, the split doses are not spread out over more than three days.

[0231] In some embodiments, the dose of cells is generally large enough to be effective in reducing disease burden.

[0232] In some embodiments, cells are administered at a desired dosage, which in some aspects includes a desired dose or number of cells or cell type(s) and / or a desired ratio of cell types. Thus, the dosage of cells is, in some embodiments, based on the total number of cells (or number per kg body weight) and the desired ratio of individual populations or subtypes, e.g., the ratio of CD4+ to CD8+. In some embodiments, the dosage of cells is based on the desired total number (or number per kg body weight) of cells in each population or of each cell type. In some embodiments, the dosage is based on a combination of such features, such as the desired number of total cells, the desired ratio, and the desired total number of cells in each population.

[0233] In some embodiments, CD8 + and CD4 +A population or subtype of cells, such as T cells, is administered at a desired dose of total cells, such as a desired dose of T cells, or within an acceptable difference. In some embodiments, the desired dose is a desired number of cells, or a desired number of cells per unit of body weight of the subject to whom the cells are administered, e.g., cells / kg. In some embodiments, the desired dose is at or exceeds a minimum number of cells or a minimum number of cells per unit of body weight. In some embodiments, among the total cells administered at a desired dose, individual populations or subtypes are administered at a desired output ratio (e.g., CD4 + Against CD8 + ) within a certain allowable difference or error in such ratio.

[0234] In some embodiments, cells are administered at a desired dose of one or more individual populations or subtypes of cells, such as a desired dose of CD4+ cells and / or a desired dose of CD8+ cells, or within an acceptable difference. In some aspects, the desired dose is the desired number of cells of a subtype or population, or the desired number of such cells per unit of body weight of the subject to whom the cells are administered, e.g., cells / kg. In some aspects, the desired dose is at or exceeds the minimum number of cells of a population or subtype, or the minimum number of cells of a population or subtype per unit of body weight.

[0235] Thus, in some embodiments, dosage is based on a desired fixed dose and desired ratio of total cells and / or based on a desired fixed dose of one or more, e.g., each, of individual subtypes or subpopulations. Thus, in some embodiments, dosage is based on a desired fixed or minimum dose of T cells and CD4 + Against CD8 + Based on the desired ratio of cells and / or CD4 + and / or CD8 + Based on desired fixation or minimum dose of cells.

[0236] In some embodiments, cells are administered at a desired output ratio or within an acceptable range of multiple cell populations or subtypes, such as CD4+ and CD8+ cells or subtypes. In some aspects, the desired ratio can be a specific ratio or a range of ratios. For example, in some embodiments, the desired ratio (e.g., CD4 + Against CD8 + The ratio of cells is between 5:1 or about to 5:1 or about to (or greater than about 1:5 and less than about 5:1), or between 1:3 or about to 3:1 or about to (or greater than about 1:3 and less than about 3:1), for example, between 2:1 or about to 1:5 or about to (or greater than about 1:5 and less than about 2:1), for example, 5:1, 4.5:1, 4:1, 3.5:1, 3:1 , 2.5:1, 2:1, 1.9:1, 1.8:1, 1.7:1, 1.6:1, 1.5:1, 1.4:1, 1.3:1, 1.2:1, 1.1:1, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5 or 1:5 or an approximation thereof. In some embodiments, acceptable differences are within about 1%, about 2%, about 3%, about 4%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50% of the desired ratio, including any value in between these ranges.

[0237] In some embodiments, the cell dose or composition comprises a defined or target ratio of CD4+ cells expressing the recombinant receptor to CD8+ cells expressing the recombinant receptor and / or CD4+ cells to CD8+ cells that is approximately 1:1 or between approximately 1:3 and approximately 3:1, e.g., approximately 1:1. In some embodiments, the cell dose or composition comprises a defined or target ratio of CD4+ cells expressing the recombinant receptor to CD8+ cells expressing the recombinant receptor and / or CD4+ cells to CD8+ cells that is approximately 1:1. In some embodiments, the cell dose or composition comprises a defined or target ratio of CD4+ cells expressing the recombinant receptor to CD8+ cells expressing the recombinant receptor and / or CD4+ cells to CD8+ cells that is between approximately 1:3 and approximately 3:1.

[0238] In certain embodiments, the number and / or concentration of cells refers to the number of recombinant receptor (e.g., CAR)-expressing cells. In other embodiments, the number and / or concentration of cells refers to the number or concentration of administered total cells, T cells, or peripheral blood mononuclear cells (PBMCs).

[0239] In some embodiments, the size of the dose is determined based on one or more criteria such as the subject's response to previous treatment, e.g., chemotherapy; the disease burden in the subject, e.g., tumor burden, volume, size or extent, degree or type of metastasis, stage; and / or the likelihood or incidence that the subject will develop a toxic outcome, e.g., CRS, macrophage activation syndrome, tumor lysis syndrome, neurotoxicity, and / or host immune response, to the cells and / or recombinant receptor being administered.

[0240] In some embodiments, for example, the dose is 5.0 x 10 6 ~2.25×10 7 , 5.0×10 6 ~2.0×10 7 , 5.0×10 6 ~1.5×10 7 , 5.0×10 6 ~1.0×10 7, 5.0×10 6 ~7.5×10 6 , 7.5×10 6 ~2.25×10 7 , 7.5×10 6 ~2.0×10 7 , 7.5×10 6 ~1.5×10 7 , 7.5×10 6 ~1.0×10 7 , 1.0×10 7 ~2.25×10 7 , 1.0×10 7 ~2.0×10 7 , 1.0×10 7 ~1.5×10 7 , 1.5×10 7 ~2.25×10 7 , 1.5×10 7 ~2.0×10 7 , 2.0×10 7 ~2.25×10 7 In some embodiments, the dose of cells contains between about 1.5 x 10 recombinant receptor-expressing cells. 8 recombinant receptor-expressing cells, approximately 3.0 x 10 8 recombinant receptor-expressing cells, or approximately 4.5 x 10 8 In some embodiments, the dose of cells contains at least or at least about 5 x 10 recombinant receptor-expressing cells, e.g., a number of cells that are recombinant receptor-expressing cells that are CD3+. In some embodiments, the dose of cells contains at least about 5 x 10 6 , 6×10 6 , 7×10 6 , 8×10 6 , 9×10 6 , 10×10 6 ~Approx. 15×10 6 In some embodiments, such a dose, e.g., such a target number of cells, refers to the total recombinant receptor-expressing cells in the administered composition.

[0241] In some embodiments, for example, the lower dose is about 5×10 per kilogram of subject body weight. 6Fewer than about 4.5 x 10 cells, recombinant receptor (e.g., CAR)-expressing cells, T cells and / or PBMCs, e.g., about 4.5 x 10 per kilogram of subject body weight 6 , 4×10 6 , 3.5×10 6 , 3×10 6 , 2.5×10 6 , 2 × 10 6 , 1.5×10 6 , 1×10 6 , 5×10 5 , 2.5×10 5 or less than 1 x 105 such cells. In some embodiments, a lower dose is about 1 x 105 per kilogram of subject body weight. 5 , 2 × 10 5 , 5×10 5 or 1×10 6 In some embodiments, such values ​​refer to the number of recombinant receptor-expressing cells; in other embodiments, they refer to the number of T cells or PBMCs or total cells administered.

[0242] In some embodiments, the subject receives multiple doses of cells, for example, two or more doses or multiple sequential doses. In some embodiments, two doses are administered to the subject. In some embodiments, the subject receives sequential doses, for example, the second dose is administered approximately 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 days after the first dose. In some embodiments, the first dose is followed by multiple sequential doses, such that the additional dose(s) are administered following the administration of the sequential doses. In some aspects, the number of cells administered to the subject in the additional doses is the same or similar to the first dose and / or the sequential doses. In some embodiments, the additional dose(s) is / are greater than the previous dose. In some embodiments, one or more subsequent doses of cells can be administered to the subject. In some embodiments, the subsequent dose of cells is administered more than or about 7, 14, 21, 28, or 35 days after the start of administration of the first dose of cells. The subsequent dose of cells can be more than, about the same as, or less than the first dose. In some embodiments, the administration of T cell therapy, such as administering the first and / or second dose of cells, may be repeated.

[0243] D. Lymphocyte Depletion Treatment In some aspects, provided methods can further include administering one or more lymphodepleting therapies, such as prior to initiating administration of T cell therapy. In some embodiments, the lymphodepleting therapy can include administration of a phosphamide, such as cyclophosphamide. In some embodiments, the lymphodepleting therapy can include administration of fludarabine.

[0244] In some embodiments, preconditioning a subject with immunodepleting (e.g., lymphodepleting) therapy can improve the efficacy of adoptive cell therapy (ACT). Preconditioning with lymphodepleting agents, including a combination of cyclosporine and fludarabine, has been effective in improving the efficacy of transferred tumor-infiltrating lymphocyte (TIL) cells in cell therapy, including improving the response and / or persistence of transferred cells. See, for example, Dudley et al., Science, 298, 850-54 (2002); Rosenberg et al., Clin Cancer Res, 17(13):4550-4557 (2011). Similarly, in the context of CAR+T cells, some studies have incorporated lymphodepleting agents, most commonly cyclophosphamide, fludarabine, bendamustine, or a combination thereof, sometimes accompanied by low-dose irradiation. Han et al. Journal of Hematology & Oncology, 6:47 (2013);Kochenderfer et al., Blood, 119: 2709-2720 (2012);Kalos et al., Sci Transl Med, 3(95):95ra73 (2011);Clinical Trial Study Record Nos.: NCT02315612; Please refer to NCT01822652.

[0245] Such preconditioning can be performed with the goal of reducing the risk of one or more of a variety of outcomes that can attenuate the effectiveness of therapy. These include the phenomenon known as "cytokine sink," in which T cells, B cells, and NK cells compete with TILs for homeostatic and activating cytokines such as IL-2, IL-7, and / or IL-15; suppression of TILs by regulatory T cells, NK cells, or other cells of the immune system; and the influence of negative regulators in the tumor microenvironment. Muranski et al., Nat Clin Pract Oncol. December; 3(12): 668-681 (2006).

[0246] Thus, in some embodiments, the provided methods further involve administering a lymphodepleting therapy to the subject. In some embodiments, the methods involve administering the lymphodepleting therapy to the subject prior to administration of the dose of cells. In some embodiments, the lymphodepleting therapy comprises a chemotherapeutic agent, such as fludarabine and / or cyclophosphamide. In some embodiments, administration of the cells and / or lymphodepleting therapy is performed by exogenous delivery.

[0247] In some embodiments, the method includes administering a preconditioning agent, such as a lymphodepleting or chemotherapeutic agent, such as cyclophosphamide, fludarabine, or a combination thereof, to the subject before administering the dose of cells. For example, the subject can be administered the preconditioning agent at least 2 days, for example, at least 3, 4, 5, 6, or 7 days, before the first or subsequent dose. In some embodiments, the subject is administered the preconditioning agent 7 days or less, for example, 6, 5, 4, 3, or 2 days or less, before administering the dose of cells.

[0248] In some embodiments, the subject is preconditioned with cyclophosphamide at a dose of 20 mg / kg to 100 mg / kg or an approximation thereof, for example, 40 mg / kg to 80 mg / kg or an approximation thereof. In some aspects, the subject is preconditioned with cyclophosphamide at 60 mg / kg or about 60 mg / kg. In some embodiments, fludarabine can be administered in a single dose or in multiple doses, for example, daily, every other day, or every three days. In some embodiments, cyclophosphamide is administered once daily for 1 or 2 days.

[0249] In some embodiments, when the lymphodepleting agent comprises fludarabine, the subject receives 1 mg / m 2 ~100mg / m 2 or any approximation thereof, e.g., 10 mg / m 2 ~75mg / m 2 , 15 mg / m2 ~50mg / m 2 20 mg / m 2 ~30mg / m 2 of or 24 mg / m 2 ~26mg / m 2 In some instances, the subject receives fludarabine at a dose of between 25 mg / m 2 In some embodiments, fludarabine is administered daily for 1-5 days, such as for 3-5 days. In some embodiments, fludarabine can be administered in a single dose, or in multiple doses, such as given daily, every other day, or every third day. In some embodiments, fludarabine is administered daily for 1-5 days, such as for 3-5 days.

[0250] In some embodiments, the lymphodepleting agent comprises a combination of drugs, such as a combination of cyclophosphamide and fludarabine. Thus, the combination of drugs can include cyclophosphamide at any dose or administration schedule, such as those described above, and fludarabine at any dose or administration schedule, such as those described above. For example, in some aspects, the subject receives cyclophosphamide at a dose of 60 mg / kg (approximately 2 g / m 2 ) of cyclophosphamide and 3 to 5 doses of 25 mg / m 2 Fludarabine is administered prior to the dose of cells.

[0251] In some embodiments, administration of a preconditioning agent prior to injection of a dose of cells improves the outcome of the treatment. For example, in some aspects, preconditioning improves the efficacy of the dose or increases the persistence of recombinant receptor-expressing cells (e.g., CAR-expressing cells, such as CAR-expressing T cells) in the subject. In some embodiments, preconditioning treatment increases disease-free survival, such as the percentage of subjects who are alive and show no minimal residual or molecularly detectable disease after a given period following a dose of cells. In some embodiments, the time to median disease-free survival is increased.

[0252] Once the cells are administered to a subject (e.g., a human), the biological activity of the engineered cell population is, in some embodiments, measured by any of a number of known methods. Parameters for evaluation include specific binding of engineered or natural T cells or other immune cells to antigens in vivo, e.g., by imaging, or ex vivo, e.g., by ELISA or flow cytometry. In certain embodiments, the ability of the engineered cells to destroy target cells can be measured using any suitable method known in the art, such as the cytotoxicity assays described in Kochenderfer et al., J. Immunotherapy, 32(7): 689-702 (2009) and Herman et al. J. Immunological Methods, 285(1): 25-40 (2004). In certain embodiments, the biological activity of the cells can also be measured by assaying the expression and / or secretion of certain cytokines, such as CD107a, IFNγ, IL-2, and TNF. In some embodiments, biological activity is measured by assessing clinical outcomes such as reduction in tumor burden or burden, hi some embodiments, toxicity outcomes, cell persistence and / or expansion, and / or the presence or absence of a host immune response are assessed.

[0253] In some embodiments, administration of a preconditioning agent prior to injection of a dose of cells improves the outcome of the treatment, such as by improving the efficacy of the dose of treatment, or increases the persistence of the recombinant receptor-expressing cells (e.g., CAR-expressing cells, such as CAR-expressing T cells) in the subject.

[0254] III. Exemplary Treatment Outcomes and Methods for Assessing Them In some embodiments of the methods, uses, kits, and articles of manufacture provided herein, the provided T cell therapy results in a characteristic associated with one or more treatment outcomes, e.g., any one or more of the parameters associated with the therapy or treatment, as described below. In some embodiments, the method includes assessing the cytotoxicity of T cells, e.g., T cells administered for a T cell-based therapy, against cancer cells. In some embodiments, the method includes assessing the exposure, persistence, and proliferation of T cells, e.g., T cells administered for a T cell-based therapy. In some embodiments, the exposure or long-term expansion and / or persistence, and / or changes in cell phenotype or functional activity of cells in the methods provided herein, e.g., cells administered for immunotherapy, e.g., T cell therapy, can be measured by assessing T cell characteristics in vivo or ex vivo. In some embodiments, such assays can be used to determine or confirm T cell, e.g., T cell therapy, function before, during, or after administration of a T cell therapy provided herein.

[0255] In some embodiments, T cell therapy may further comprise one or more screening steps to identify subjects for treatment with T cell therapy and / or continue T cell therapy, and / or to assess the outcome of the treatment and / or to monitor the outcome of the treatment. In some embodiments, the step for assessing the treatment may comprise a step for assessing and / or monitoring the treatment and / or to identify subjects for administration of further or remaining steps of therapy and / or for repeat therapy. In some embodiments, the screening step and / or assessment of the outcome of the treatment can be used to determine the dose, frequency, duration, timing, and / or sequence of the T cell therapies provided herein.

[0256] In some embodiments, any of the screening steps and / or assessments of treatment outcome described herein can be used before, during, during, or following administration of one or more steps of a provided T cell therapy (e.g., anti-BCMA CAR T cells). In some embodiments, the assessment is performed before, during, during, or following performance of any of the methods provided herein. In some embodiments, the assessment is performed before performance of a method provided herein. In some embodiments, the assessment is performed after performance of one or more steps of a method provided herein. In some embodiments, the assessment is performed before administration of one or more steps of a provided T cell therapy, e.g., to screen and identify patients suitable for and / or susceptible to receiving T cell therapy. In some embodiments, the assessment is performed during, during, or following administration of one or more steps of a provided T cell therapy, e.g., to evaluate intermediate or final treatment outcomes, e.g., to determine the effectiveness of treatment, and / or to decide whether to continue or repeat treatment, and / or to decide whether to administer remaining steps of T cell therapy.

[0257] In some embodiments, the outcome of the treatment includes an improvement in immune function, e.g., of T cells administered for cell-based therapy and / or of endogenous T cells in the body. In some embodiments, exemplary treatment outcomes include, but are not limited to, enhanced T cell proliferation, enhanced T cell functional activity, and changes in immune cell phenotypic marker expression, e.g., such characteristics are associated with engineered T cells, e.g., CAR-T cells, administered to a subject. In some embodiments, exemplary treatment outcomes include a reduction in disease burden, e.g., tumor burden, improved clinical outcome, and / or enhanced efficacy of therapy.

[0258] In some embodiments, the screening step and / or evaluation of treatment outcome comprises assessing the survival and / or function of T cells administered for cell-based therapy. In some embodiments, the screening step and / or evaluation of treatment outcome comprises assessing cytokine or growth factor levels. In some embodiments, the screening step and / or evaluation of treatment outcome comprises assessing disease burden and / or improvement, e.g., assessing tumor burden and / or clinical outcome. In some embodiments, any of the screening steps and / or evaluation of treatment outcome may include any of the evaluation methods and / or assays described herein and / or known in the art, and may be performed one or more times, e.g., before, during, during, or following administration of one or more steps of T cell therapy. Exemplary parameter sets associated with treatment outcome that can be assessed in some embodiments of the methods provided herein include peripheral blood immune cell population profiles and / or tumor burden.

[0259] In some embodiments, the method affects the efficacy of cell therapy in a subject. In some embodiments, the cytotoxicity of recombinant receptor-expressing, e.g., CAR-expressing, cells after administration of a dose of cells in a method with debulking is higher than that achieved by a method without debulking. In some embodiments, the cytotoxicity of recombinant receptor-expressing, e.g., CAR-expressing, cells after administration of a dose of cells in a method in which a subject is selected for treatment as having a serum sBCMA level below about 600 ng / mL and / or not having the presence of IgG heavy chain disease (HCD) is higher than that achieved by a method in which the subject is not selected. In some embodiments, the cytotoxicity in a subject administered T cell therapy, e.g., CAR-expressing T cells, is evaluated compared to a method in which T cell therapy is administered to a subject not selected for treatment. In some embodiments, the method results in the administered T cells exhibiting increased or prolonged cytotoxicity in the subject compared to a method in which T cell therapy is administered to a subject not selected for treatment.

[0260] In some embodiments, debulking a tumor before treatment with T cell therapy reduces the disease burden, e.g., tumor burden, in a subject compared to methods in which the tumor is not debulked before treatment. In some embodiments, selecting a subject for treatment reduces the disease burden, e.g., tumor burden, in a subject compared to methods in which the subject is not selected for treatment. In some embodiments, debulking a tumor before treatment with T cell therapy results in improved clinical outcomes, e.g., objective response rate (ORR), progression-free survival (PFS), and overall survival (OS), compared to methods in which the tumor is not debulked before treatment. In some embodiments, selecting a subject for treatment with T cell therapy results in improved clinical outcomes, e.g., objective response rate (ORR), progression-free survival (PFS), and overall survival (OS), compared to methods in which the subject is not selected for treatment.

[0261] In some embodiments, subjects can be screened before administering one or more steps of T cell therapy. For example, subjects can be screened for characteristics of the disease and / or disease burden, such as tumor burden, before administering T cell therapy to determine suitability, responsiveness, and / or sensitivity to the administration of T cell therapy. For example, subjects can be screened for characteristics of the disease before administering T cell therapy to determine suitability, responsiveness, and / or sensitivity to the administration of T cell therapy. In some embodiments, screening steps and / or evaluation of treatment outcomes can be used to determine the dose, frequency, duration, timing, and / or sequence of the T cell therapy provided herein.

[0262] In some embodiments, subjects may be screened after administration of one of the steps of T cell therapy to determine and identify subjects to receive the remaining steps of T cell therapy and / or to monitor the effectiveness of the therapy. In some embodiments, the number, level or amount of administered T cells and / or the proliferation and / or activity of administered T cells is assessed after administration of the engineered T cells.

[0263] In some embodiments, changes and / or alterations, e.g., increases, elevations, decreases, or reductions, in the level, value, or measurement of a parameter or outcome are determined or assessed compared to the level, value, or measurement of the same parameter or outcome at different time points of assessment, under different conditions, at a reference time point, and / or in different subjects. In some embodiments, the levels, values, or measurements of two or more parameters are determined, and the relative levels are compared. In some embodiments, the determined level, value, or measurement of a parameter is compared to the level, value, or measurement from a control sample or an untreated sample. In some embodiments, the determined level, value, or measurement of a parameter is compared to the level from a sample from the same subject but at a different time point. The values ​​obtained by quantifying individual parameters can be combined for the purpose of disease assessment by performing arithmetic or logical operations on the parameter levels, values, or measurements, for example, by using multiparametric analysis. In some embodiments, the ratio of two or more specific parameters can be calculated.

[0264] Assessment and determination of parameters related to T cell health, function, activity, and / or outcome, e.g., response, efficacy, and / or toxicity outcomes, can be assessed at various time points, hi some embodiments, assessment can be performed multiple times, e.g., before, during, and / or after manufacturing of the cells, before, during, and / or after initiation of administration of T cell therapy.

[0265] In some embodiments, functional attributes of the administered cells and / or cell compositions include pharmacokinetic (PK) and pharmacodynamic parameters, cell expansion and persistence, cell function assays (e.g., cytotoxicity assays, cytokine secretion assays, and in vivo assays, any of those described herein), high-dimensional T cell signaling assessment, and assessment of T cell exhaustion phenotype and / or signature.

[0266] In some embodiments, parameters associated with the outcome of a therapy or treatment, including parameters that can be evaluated for screening and / or evaluation of the outcome of a treatment and / or to monitor the outcome of a treatment, include tumor or disease burden.Administering immunotherapy, such as T cell therapy (e.g., CAR-expressing T cells), can reduce or prevent the growth or burden of a disease or condition in a subject.For example, if the disease or condition is a tumor, the method generally reduces tumor size, bulk, metastasis, the percentage of blasts in the bone marrow, or molecularly detectable cancer, and / or improves prognosis or survival or other symptoms associated with tumor burden.

[0267] In some embodiments, the provided methods result in a reduced tumor burden in a treated subject compared to alternative methods in which T cell therapy (e.g., anti-BCMA CAR T cells) is administered without debulking the tumor prior to treatment. In some embodiments, the provided methods result in a reduced tumor burden in a treated subject compared to alternative methods in which T cell therapy (e.g., anti-BCMA CAR T cells) is administered without selecting the subject for treatment.

[0268] Tumor burden need not actually be reduced in all subjects receiving T cell therapy, e.g., based on clinical data, on average in treated subjects, where the tumor burden is such that a majority of subjects treated with such T cell therapy exhibit a reduction in tumor burden, e.g., at least 50%, 60%, 70%, 80%, 90%, 95% or more of subjects treated with T cell therapy exhibit a reduction in tumor burden.

[0269] Disease burden can include the total number of diseased cells in a subject, or in a subject's organs, tissues, or bodily fluids, such as tumors, or organs or tissues in other locations, such as those that represent metastases. For example, tumor cells can be detected and / or quantified in the blood, lymph, or bone marrow in the setting of certain hematological malignancies. Disease burden, in some embodiments, can include tumor mass, the number or extent of metastases, and / or the percentage of blasts present in the bone marrow.

[0270] In the case of MM, exemplary parameters for assessing the degree of disease burden include the number of clonal plasma cells (e.g., greater than 10% in bone marrow biopsy or any amount in biopsies from other tissues; plasmacytoma), the presence of a monoclonal protein (paraprotein) in either serum or urine, evidence of end-organ damage felt to be related to plasma cell dyscrasia (e.g., hypercalcemia (corrected calcium greater than 2.75 mmol / l); renal failure due to myeloma; anemia (hemoglobin less than 10 g / dl); and / or bone lesions (lytic lesions or osteoporosis with compression fractures)).

[0271] Exemplary methods for assessing disease status or disease burden include measuring M protein in biological fluids such as blood and / or urine by electrophoresis and immunofixation; quantifying sFLC (κ and λ) in blood; skeletal examination; and imaging by positron emission tomography (PET) / computed tomography (CT) in subjects with extramedullary disease. In some embodiments, disease status can be assessed by bone marrow examination. In some examples, the efficacy of T cell therapy following its administration to a subject is determined by the expansion and persistence of T cells (e.g., BCMA CAR cells) in the blood and / or bone marrow. In some embodiments, the efficacy of T cell therapy is determined based on the anti-tumor activity of the administered cells (e.g., BCMA CAR T cells). In some embodiments, anti-tumor activity is determined by overall response rate (ORR) and / or the International Myeloma Working Group (IMWG) Unified Response Criteria (see Kumar et al. (2016) Lancet Oncol 17(8):e328-346). In some embodiments, response is assessed using minimal residual disease (MRD) assessment. In some embodiments, MRD can be assessed by methods such as flow cytometry and high-throughput sequencing, e.g., deep sequencing. In some aspects, subjects with MRD-negative disease are assessed by flow cytometry (next-generation flow cytometry; NGF) or high-throughput sequencing, e.g., deep sequencing or next-generation sequencing (NGS), for example. 5 1 minimum sensitivity in ≥ 10 nucleated cells (i.e., 10 -5 Subjects who demonstrate the absence of abnormal clonal plasma cells in the bone marrow aspirate as excluded by an assay with a sensitivity (sensitivity) of 100%.

[0272] In some embodiments, sustained MRD negativity includes subjects who demonstrate MRD negativity in bone marrow (NGF and / or NGS) and by imaging, as defined below, confirmed at a minimum of one year apart. Subsequent evaluations can be used to further specify duration of negativity (e.g., MRD negativity at 5 years). In some embodiments, flow MRD negativity is measured at 10 5 In some embodiments, sequencing MRD negative includes subjects who demonstrate the absence of phenotypically abnormal clonal plasma cells by NGS in bone marrow aspirate using the EuroFlow standard operating procedure for MRD detection in multiple myeloma (or a validated equivalent method) with a minimum sensitivity of 1 in 10 or more nucleated cells. In some embodiments, sequencing MRD negative includes subjects who demonstrate the absence of clonal plasma cells by NGS in bone marrow aspirate, where the presence of a clone is greater than 10 5 This is defined as fewer than two identical sequencing reads obtained after DNA sequencing of bone marrow aspirate using the LymphoSIGHT platform (or a validated equivalent method) with a minimum sensitivity of 1 in nucleated cells or more. In some embodiments, imaging plus MRD negativity includes subjects who exhibit MRD negativity as assessed by NGF or NGS plus a total loss of elevated tracer uptake seen on baseline or prior PET / CT, or a decrease to less than mediastinal blood pool SUV or surrounding normal tissue (see Kumar et al. (2016) Lancet Oncol 17(8):e328-346).

[0273] In some aspects, the subject's survival, survival within a certain period of time, degree of survival, the presence or duration of event-free or symptom-free survival, or recurrence-free survival are evaluated. In some embodiments, any symptom of the disease or condition is evaluated. In some embodiments, a measure of tumor burden is specified. In some embodiments, exemplary parameters for determination include specific clinical outcomes indicating remission or improvement in tumor. Such parameters include duration of disease control, including objective response (OR), complete response (CR), stringent complete response (sCR), very good partial response (VGPR), partial response (PR), minimal response (MR), stable disease (SD), progressive disease (PD), or relapse (see, e.g., International Myeloma Working Group (IMWG) Unified Response Criteria; see Kumar et al. (2016) Lancet Oncol 17(8):e328-346), objective response rate (ORR), progression-free survival (PFS), and overall survival (OS). In some embodiments, response is evaluated using minimal residual disease (MRD) assessment. In some embodiments, response is assessed using a complete response (CR) or stringent complete response (sCR) assessment. In some embodiments, response is assessed using a complete response (CR) assessment. In some embodiments, response is assessed using a stringent CR assessment. Specific thresholds for parameters can be set to determine the effectiveness of the methods provided herein. In some embodiments, the disease or disorder to be treated is multiple myeloma. In some embodiments, the criteria for measurable disease for multiple myeloma can include: (1) serum M protein ≥ 1 g / dL; (2) urinary M protein ≥ 200 mg / 24 hours; (3) diseased serum free light chain (sFLC) levels ≥ 10 mg / dL and an abnormal κ to λ ratio. In some cases, light chain disease is only permitted in subjects without measurable disease in serum or urine.

[0274] In some embodiments, the response is assessed based on the duration of response following administration of T cell therapy, e.g., BCMA CAR T cells. In some aspects, for example, response to therapy according to provided embodiments can be measured at a designated time point after initiation of administration of T cell therapy. In some embodiments, the designated time point is at or about 1, 2, 3, 6, 9, 12, 18, 24, 30, or 36 months after initiation of administration, or within a range defined by any of the foregoing. In some embodiments, the designated time point is at or about 4, 8, 12, 16, 20, 24, 28, 32, 36, 48, or 52 weeks or months after initiation of administration, or within a range defined by any of the foregoing. In some embodiments, the designated time point is at or about 1 month after initiation of administration. In some embodiments, the designated time point is at or about 3 months after initiation of administration. In some embodiments, the designated time point is at or about 6 months after initiation of administration. In some embodiments, the designated time point is at or about 9 months after initiation of administration. In some embodiments, the specified time point is 12 months or approximately 12 months after the start of administration. In some embodiments, the response is CR or sCR. In some embodiments, the response is CR. In some embodiments, the response is sCR.

[0275] In some embodiments, the response or outcome determined at or about 3, 6, 9, or 12 months after a specified time point is equivalent to or improved compared to the response or outcome determined at the earlier specified time point. For example, in some aspects, if the response or outcome determined at the earlier specified time point is stable disease (SD), progressive disease (PD), or relapse, subjects treated according to provided embodiments may exhibit an equivalent or improved response or outcome at a subsequent time point at or about 3, 6, 9, or 12 months after the earlier specified time point, which is equivalent to the response or outcome at the earlier specified time point, or a response or outcome that is an objective response (OR), complete response (CR), stringent complete response (sCR), very good partial response (VGPR), or partial response (PR) (e.g., exhibiting a better response outcome according to the International Myeloma Working Group (IMWG) Uniform Response Criteria; see Kumar et al. (2016) Lancet Oncol 17(8):e328-346). In some embodiments, the response is CR or sCR. In some embodiments, the response is CR. In some embodiments, the response is sCR. In some aspects, subjects treated according to provided embodiments can exhibit an improved response or outcome between two decision time points. In some aspects, a subject can exhibit PR or VGPR at an initial designated time point for evaluation, for example, 4 weeks after initiation of administration, and then exhibit an improved response, such as CR or sCR, at a later time point, for example, 12 weeks after initiation of administration. In some aspects, progression-free survival (PFS) is described as the period during and after treatment of a disease, such as cancer, during which a subject survives with the disease but does not worsen. In some aspects, objective response (OR) is described as a measurable response. In some aspects, objective response rate (ORR; in some cases, also known as overall response rate) is described as the percentage of patients who achieve CR or PR. In some aspects, overall survival (OS) is described as the length of time from the date of diagnosis or initiation of treatment of a disease, such as cancer, during which a subject diagnosed with the disease is still alive.In some embodiments, event-free survival (EFS) is described as the period after completion of treatment for cancer during which a subject remains free of certain complications or events that the treatment was intended to prevent or delay. These events may include cancer recurrence or the onset of certain symptoms, such as bone pain from cancer that has spread to the bone, or death.

[0276] In some embodiments, measuring duration of response (DOR) includes the time from documenting tumor response to disease progression. In some embodiments, parameters for assessing response may include a sustained response, for example, a response that persists after a period of time from the initiation of therapy. In some embodiments, a sustained response is indicated by a response rate approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, or 24 months after the initiation of therapy. In some embodiments, the response or outcome is durable for more than 3, 6, 9, or 12 months or about that number.

[0277] In some embodiments, the Eastern Cooperative Oncology Group (ECOG) performance status index can be used to evaluate or select subjects for treatment, for example, subjects who have had poor results from previous therapy (see, e.g., Oken et al. (1982) Am J Clin Oncol. 5:649-655). The ECOG performance status scale describes a patient's level of function in terms of their ability to manage themselves, their daily activities, and their physical abilities (e.g., walking, working, etc.). In some embodiments, an ECOG performance status of 0 indicates that the subject can perform normal activities. In some aspects, a subject with an ECOG performance status of 1 shows some limitations in physical activity, but the subject is fully ambulatory. In some aspects, a patient with an ECOG performance status of 2 is more than 50% ambulatory. In some cases, a subject with an ECOG performance status of 2 can also be capable of self-care; see, e.g., Sorensen et al., (1993) Br J Cancer 67(4) 773-775. In some embodiments, subjects to be administered according to the methods or treatment regimens provided herein include subjects with an ECOG performance status of 0 or 1.

[0278] In some embodiments, the method and / or administration of the T cell therapy (e.g., BCMA CAR T cells) reduces the disease burden compared to the disease burden at a time point immediately prior to administration of the T cell therapy.

[0279] In some aspects, administration of T cell therapy may prevent an increase in disease burden, which may be manifested by no change in disease burden.

[0280] In some embodiments, the method reduces disease or symptom burden, e.g., tumor cell count, tumor size, duration of patient survival, or event-free survival, to a greater extent and / or for a longer duration when compared to the reduction that would be observed with a comparable method using an alternative therapy, e.g., one in which the subject receives T cell therapy in the absence of tumor debulking prior to treatment and / or when the subject is not selected prior to treatment. In some embodiments, disease burden is reduced to a greater extent or for a longer duration following administration of T cell therapy compared to the reduction that would result without tumor debulking prior to treatment and / or without patient selection for treatment.

[0281] In some embodiments, the burden of a disease or condition in a subject is detected, assessed, or measured. In some aspects, the disease burden can be detected by detecting the total number of disease or disease-related cells, e.g., tumor cells, in a subject or in the subject's organs, tissues, or bodily fluids, e.g., blood or serum. In some embodiments, the disease burden, e.g., tumor burden, is assessed by measuring the mass of a solid tumor and / or the number or extent of metastases. In some aspects, the subject's survival, survival within a certain period of time, degree of survival, presence or duration of event-free or symptom-free survival, or recurrence-free survival is assessed. In some embodiments, any symptom of the disease or condition is assessed. In some embodiments, a measure of the burden of the disease or condition is specified. In some embodiments, exemplary parameters for determination include specific clinical outcomes indicative of regression or improvement of a disease or condition, e.g., a tumor. Such parameters include complete response (CR), partial response (PR), or stable disease (SD) (see, e.g., Response Evaluation Criteria In Solid Tumors (RECIST) guidelines), objective response rate (ORR), duration of disease control, including progression-free survival (PFS), and overall survival (OS). In some embodiments, the parameter is CR or sCR. In some embodiments, the parameter is CR. In some embodiments, the parameter is sCR. Specific thresholds for the parameters can be set to determine the efficacy of the methods of T cell therapy provided herein.

[0282] In some embodiments, subjects treated according to the method achieve a more durable response. In some cases, the measure of duration of response (DOR) includes the time from documentation of tumor response to disease progression. In some embodiments, parameters for assessing response can include a durable response, e.g., a response that persists after a period from initiation of therapy. In some embodiments, a durable response is indicated by a response rate approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, or 24 months after initiation of therapy. In some embodiments, the response is durable for more than 3 months, more than 6 months, or more than 12 months. In some specific embodiments, subjects treated according to the method achieve a more durable response after the subject previously relapsed following remission in response to administration of the genetically engineered cells.

[0283] In some aspects, disease burden is measured or detected prior to administration of debulking, prior to administration of T cell therapy, and / or after debulking but prior to administration of T cell therapy. In the context of multiple administrations of one or more steps of T cell therapy, disease burden may in some embodiments be measured prior to, following, or at a time point between administrations of any of the steps, doses, and / or cycles of administration.

[0284] In some embodiments, the burden is reduced by the provided methods by, or by at least, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 percent, or by at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 percent, compared to immediately prior to administration of the T cell therapy. In some embodiments, the disease burden, tumor size, tumor volume, tumor mass, and / or tumor burden or bulk is reduced following administration of the T cell therapy and debulking by at least, 20, 30, 40, 50, 60, 70, 80, 90, or more percent, or by at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, or more percent, compared to immediately prior to administration of the T cell therapy and / or debulking.

[0285] In some embodiments, reducing disease burden with the method includes, e.g., inducing a complete morphological response when assessed 1 month, 2 months, 3 months, or more than 3 months after administration, e.g., initiation, of T cell therapy.

[0286] In some embodiments, the assay for minimal residual disease is negative, e.g., as measured by multiparametric flow cytometry, or the level of minimal residual disease is less than about 0.3%, less than about 0.2%, less than about 0.1%, or less than 0.05%.

[0287] In some embodiments, the subject's event-free survival or overall survival rate is improved by the method compared to other methods. For example, in some embodiments, the event-free survival rate or probability for subjects treated with the method of T cell therapy provided herein 6 months following the method is greater than about 40%, greater than about 50%, greater than about 60%, greater than about 70%, greater than about 80%, greater than about 90%, or greater than about 95%. In some aspects, the overall survival rate is greater than about 40%, greater than about 50%, greater than about 60%, greater than about 70%, greater than about 80%, greater than about 90%, or greater than about 95%. In some embodiments, subjects treated with the method exhibit event-free survival, relapse-free survival, or survival for at least 6 months, or for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years. In some embodiments, the time to progression is improved, for example, more than 6 months or at least more than or about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years.

[0288] In some embodiments, following treatment with the method, the likelihood of relapse is reduced as compared to other methods, for example, in some embodiments, the likelihood of relapse 6 months after the method of T cell therapy is less than about 80%, less than about 70%, less than about 60%, less than about 50%, less than about 40%, less than about 30%, less than about 20%, or less than about 10%.

[0289] In some embodiments, administration can treat a subject despite the subject becoming refractory to another therapy. In some embodiments, when administered to a subject in accordance with the embodiments described herein, a dose or composition can achieve a complete response (CR) or severe CR (sCR) in at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the subjects to which it is administered. In some embodiments, when administered to a subject in accordance with the embodiments described herein, a dose or composition can achieve a complete response (CR) in at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the subjects to which it is administered. In some embodiments, when administered to a subject in accordance with the embodiments described herein, a dose or composition can achieve a severe complete response (sCR) in at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the subjects to which it is administered. In some embodiments, when administered to a subject according to the embodiments described herein, a dose or composition can achieve an objective response (OR) in at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of administered subjects. In some embodiments, OR includes subjects achieving a stringent complete response (sCR), a complete response (CR), a very good partial response (VGPR), a partial response (PR), and a minimal response (MR). In some embodiments, when administered to a subject according to the embodiments described herein, a dose or composition can achieve a stringent complete response (sCR), a complete response (CR), a very good partial response (VGPR), or a partial response (PR) in at least 50%, 60%, 70%, 80%, or 85% of administered subjects. In some embodiments, when administered to a subject according to the embodiments described herein, a dose or composition can achieve a stringent complete response (sCR) or a complete response (CR) in at least 20%, 30%, 40%, 50%, 60%, or 70% of administered subjects.In some embodiments, when administered to a subject according to the embodiments described herein, the dose or composition is capable of achieving a stringent complete response (sCR) in at least 20%, 30%, 40%, 50%, 60%, or 70% of the subjects to which it is administered. In some embodiments, when administered to a subject according to the embodiments described herein, the dose or composition is capable of achieving a complete response (CR) in at least 20%, 30%, 40%, 50%, 60%, or 70% of the subjects to which it is administered. In some embodiments, an exemplary dose is about 1.0 x 10. 7 , 1.5×10 7 , 2.0×10 7 , 2.5×10 7 , 5.0×10 7 , 1.5×10 8 , 3.0×10 8 , 4.5×10 8 , 6.0×10 8 , or 8 × 10 8 In some embodiments, for example, a specific response to treatment according to the methods provided herein can be assessed based on the International Myeloma Working Group (IMWG) Unified Response Criteria (see Kumar et al. (2016) Lancet Oncol 17(8):e328-346).

[0290] IV. BCMA-Targeted Therapies and Engineered Cells Also provided herein are BCMA-targeted therapies. In some embodiments, the BCMA-targeted therapy is provided to a subject with multiple myeloma. In some embodiments, the BCMA-targeted therapy is an antibody, an antibody-drug conjugate (ADC), or a T cell engager. In some embodiments, the BCMA-targeted therapy is an antibody. In some embodiments, the BCMA-targeted therapy is an antibody-drug conjugate (ADC). In some embodiments, the BCMA-targeted therapy is a T cell engager (TCE). In some embodiments, the BCMA-targeted therapy is a T cell engager capable of stimulating T cell activity. In some embodiments, the BCMA-targeted therapy is a bispecific T cell engager (BiTE) therapy. In some embodiments, the BCMA-targeted therapy is a cell therapy selected from tumor-infiltrating lymphocyte (TIL) therapy, endogenous T cell therapy, natural killer (NK) cell therapy, transgenic TCR therapy, and recombinant receptor-expressing cell therapy, and may be chimeric antigen receptor (CAR)-expressing cell therapy. In some embodiments, the cell therapy is a recombinant receptor-expressing cell therapy. In some embodiments, the cell therapy is a chimeric antigen receptor (CAR)-expressing cell therapy. In any of the embodiments provided herein, a BCMA-targeted therapy is administered to a subject with multiple myeloma.

[0291] Also provided herein are cells, such as engineered cells, containing a recombinant receptor (e.g., a chimeric antigen receptor), such as one that contains an extracellular domain comprising an anti-BCMA binding moiety, such as an antibody or fragment described herein. Also provided are populations of such cells, compositions containing such cells, and / or compositions enriched for such cells, wherein the cells expressing a BCMA binding molecule constitute at least 50, 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or more percent of the total cells in the composition or of cells of a particular type, such as PBMCs, T cells, or CD3+, CD8+, or CD4+ cells.

[0292] Among the compositions, pharmaceutical compositions and formulations for administration, such as for adoptive cell therapy, are particularly mentioned. Methods of therapy for administering the cells and compositions to a subject, e.g., a patient, are also provided.

[0293] Thus, genetically engineered cells expressing recombinant receptors containing antibodies, such as cells containing CARs, are also provided. The cells are generally eukaryotic cells, such as mammalian cells, and usually human cells. In some embodiments, the cells are cells of the immune system, such as myeloid or lymphoid cells, derived from blood, bone marrow, lymph, or lymphoid organs, and include lymphocytes, usually T cells and / or NK cells, including cells of the innate or adaptive immune system. Other exemplary cells include stem cells, such as multipotent and pluripotent stem cells, including induced pluripotent stem cells (iPSCs). The cells are usually primary cells, such as those isolated directly from a subject and / or isolated and frozen from a subject. In some embodiments, the cells include one or more subsets of T cells or other cell types, e.g., the total T cell population, CD4+ cells, CD8+ cells, and subpopulations thereof, e.g., those defined by function, activation state, maturity, differentiation potential, expansion, recirculation, localization, and / or persistence, antigen specificity, antigen receptor type, presence in specific organs or compartments, marker or cytokine secretion profile, and / or degree of differentiation. With respect to the subject to be treated, the cells can be allogeneic and / or autologous. Some methods include commercially available methods. In some aspects, e.g., with commercially available technologies, the cells are pluripotent and / or multipotent, e.g., stem cells, e.g., induced pluripotent stem cells (iPSCs). In some embodiments, the methods include isolating cells from a subject, preparing, processing, culturing, and / or manipulating them, and reintroducing them into the same patient before or after cryopreservation.

[0294] Among the subtypes and subpopulations of T cells and / or CD4+ and / or CD8+ T cells are naive T (TN) cells, effector T cells (TEFF), memory T cells and their subtypes, such as stem cell memory T (TSCM), central memory T (TCM), effector memory T (TEM), or well-differentiated effector memory T cells, cell-infiltrating lymphocytes (TIL), immature T cells, mature T cells, helper T cells, cytotoxic T cells, mucosal-associated invariant T (MAIT) cells, naturally occurring adaptive regulatory T (Treg) cells, helper T cells, such as TH1 cells, TH2 cells, TH3 cells, TH17 cells, TH9 cells, TH22 cells, follicular helper T cells, alpha / beta T cells, and delta / gamma T cells.

[0295] In some embodiments, the cells are natural killer (NK) cells. In some embodiments, the cells are monocytes or granulocytes, such as myeloid cells, macrophages, neutrophils, dendritic cells, mast cells, eosinophils, and / or basophils.

[0296] In some embodiments, the cells contain one or more polynucleotides introduced by genetic engineering, thereby expressing recombinant or genetically engineered products of such polynucleotides. In some embodiments, the polynucleotides are heterologous, i.e., not normally present in the cell or sample obtained from the cell, e.g., obtained from another organism or cell not normally found in the engineered cell and / or the organism from which such cell is derived. In some embodiments, the polynucleotides are non-naturally occurring, e.g., polynucleotides not found in nature, including those that contain chimeric combinations of polynucleotides encoding various domains from multiple different cell types. In some embodiments, the cells (e.g., engineered cells) contain a vector (e.g., a viral vector, an expression vector, etc.) described herein, e.g., a vector comprising a nucleic acid encoding a recombinant receptor described herein.

[0297] In some embodiments, T cell therapy for use in conjunction with the provided methods involves administering engineered T cells expressing a recombinant receptor designed to recognize and / or specifically bind to a molecule (e.g., BCMA) associated with multiple myeloma, e.g., relapsed and refractory (R / R) multiple myeloma (MM). In some embodiments, binding to the antigen results in a response, such as an immune response, to such a molecule after binding to the molecule. In some embodiments, the cells contain or are engineered to contain an engineered receptor, e.g., an engineered antigen receptor, e.g., a chimeric antigen receptor (CAR) or T cell receptor (TCR). Recombinant receptors, such as CARs, generally comprise an extracellular antigen (or ligand) binding domain directed against BCMA, linked, in some aspects, via a linker and / or transmembrane domain(s), to one or more intracellular signaling components. In some aspects, the engineered cells are provided in pharmaceutical compositions and formulations suitable for administration to a subject, such as for adoptive cell therapy. Methods of therapy for administering the cells and compositions to a subject, e.g., a patient, are also provided.

[0298] In some embodiments, the cells contain one or more nucleic acids that have been introduced by genetic engineering, thereby expressing recombinant or engineered products of such nucleic acids. In some embodiments, gene transfer is achieved by first stimulating the cells, e.g., in combination with a stimulus that induces a response such as proliferation, survival, and / or activation, e.g., as measured by expression of cytokines or activation markers, and then transducing and expanding the activated cells in culture to numbers sufficient for clinical application.

[0299] A. Recombinant Receptors, e.g., Chimeric Antigen Receptors (CARs) The cells generally express recombinant receptors, such as antigen receptors, including functional non-TCR antigen receptors, other antigen-binding receptors, such as chimeric antigen receptors (CARs), transgenic T cell receptors (TCRs), and other chimeric receptors.

[0300] In some embodiments of the provided methods and uses, engineered cells, such as T cells, express a chimeric receptor, such as a chimeric antigen receptor (CAR), that contains one or more domains that combine a ligand-binding domain (e.g., an antibody or antibody fragment) that provides specificity for a desired antigen (e.g., a tumor antigen) with an intracellular signaling domain. In some embodiments, the intracellular signaling domain is an activation intracellular domain portion, such as a T cell activation domain, that provides a primary activation signal. In some embodiments, the intracellular signaling domain contains, or additionally contains, a costimulatory signaling domain to facilitate effector function. After specific binding to a molecule, e.g., an antigen, the receptor generally delivers an immunostimulatory signal, such as an ITAM transduction signal, to the cell, thereby promoting an immune response targeted to the disease or condition. In some embodiments, the chimeric receptor, when engineered into an immune cell, can modulate T cell activity, and in some cases, modulate T cell differentiation or homeostasis, resulting in engineered cells with improved longevity, survival, and / or persistence in vivo, such as for use in adoptive cell therapy methods.

[0301] In some embodiments, CARs are constructed with specificity for a particular antigen (or marker or ligand), such as an antigen expressed in a particular cell type to be targeted by adoptive therapy, e.g., a cancer marker, and / or an antigen expressed in a normal or non-diseased cell type, against which an attenuating response is intended to be induced. Thus, a CAR typically comprises, in its extracellular portion, one or more antigen-binding molecules, e.g., one or more antigen-binding fragments, domains, or portions, or one or more antibody variable domains and / or antibody molecules.

[0302] The term "antibody" is used herein in the broadest sense and includes antigen-binding fragments (Fab) fragments, F(ab')2 fragments, Fab' fragments, Fv fragments, recombinant IgG (rIgG) fragments, heavy chain variable (V) fragments capable of specifically binding to an antigen, and H (scFv) regions, single-chain antibody fragments including single-chain variable fragments (scFv), and single-domain antibody (e.g., sdAb, sdFv, nanobody, VHH) fragments. The term encompasses genetically engineered and / or otherwise modified forms of immunoglobulins, such as intrabodies, peptibodies, chimeric antibodies, fully human antibodies, humanized antibodies, and heteroconjugate antibodies, multispecific, e.g., bispecific or trispecific, antibodies, diabodies, triabodies, and tetrabodies, tandem di-scFv, tandem tri-scFv. Unless otherwise stated, the term "antibody" should be understood to encompass functional antibody fragments thereof, also referred to herein as "antigen-binding fragments." The term also encompasses intact or whole antibodies, including antibodies of any class or subclass, including IgG and its subclasses, IgM, IgE, IgA, and IgD.

[0303] The terms "complementarity-determining region" and "CDR," synonymous with "hypervariable region" or "HVR," are known in the art to refer to non-contiguous sequences of amino acids in an antibody variable region that confer antigen specificity and / or binding affinity. Generally, there are three CDRs (CDR-H1, CDR-H2, CDR-H3) in each heavy chain variable region and three CDRs (CDR-L1, CDR-L2, CDR-L3) in each light chain variable region. The terms "framework region" and "FR" are known in the art to refer to the non-CDR portions of the heavy and light chain variable regions. Generally, there are four FRs (FR-H1, FR-H2, FR-H3, and FR-H4) in each full-length heavy chain variable region and four FRs (FR-L1, FR-L2, FR-L3, and FR-L4) in each full-length light chain variable region.

[0304] The precise amino acid sequence boundaries of a given CDR or FR can be determined using the Kabat numbering scheme (Kabat et al., 1991), "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD; Al-Lazikani et al., (1997) JMB 273,927-948 (Chothia numbering scheme); MacCallum et al., J. Mol. Biol. 262:732-745 (1996), "Antibody-antigen interactions: Contact analysis and binding site topography," J. Mol. Biol. 262, 732-745 (Contact numbering scheme); Lefranc MP et al., "IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains," Dev Comp Immunol, 2003. Jan;27(1):55-77 (the "IMGT" numbering scheme); Honegger A and Plueckthun A, "Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool," J Mol Biol, 2001 Jun 8;309(3):657-70 (the "Aho" numbering scheme); and Martin et al., "Modeling antibody hypervariable loops: a combined algorithm," PNAS, 1989, 86(23):9268-9272 (the "AbM" numbering scheme).

[0305] The boundaries of a given CDR or FR may vary depending on the scheme used for identification. For example, the Kabat scheme is based on structural alignment, while the Chothia scheme is based on structural information. The numbering for both the Kabat and Chothia schemes is based on the length of the most common antibody region sequences, with insertions adjusted by an insertion character, e.g., "30a," and deletions occurring in some antibodies. The two schemes place certain insertions and deletions ("indels") in different positions, resulting in differential numbering. The contact scheme is based on the analysis of complex crystal structures and is similar in many respects to the Chothia numbering scheme. The AbM scheme is a compromise between the Kabat and Chothia definitions, based on those used by Oxford Molecular's AbM antibody modeling software.

[0306] Table 2 below lists exemplary boundary positions for CDR-L1, CDR-L2, CDR-L3, and CDR-H1, CDR-H2, and CDR-H3, as identified by the Kabat, Chothia, AbM, and contact schemes, respectively. For CDR-H1, residue numbering is listed using both the Kabat and Chothia numbering schemes. FRs are positioned between the CDRs; for example, FR-L1 precedes CDR-L1, FR-L2 is positioned between CDR-L1 and CDR-L2, FR-L3 is positioned between CDR-L2 and CDR-L3, etc. Note that the Kabat numbering scheme shown places insertions at H35A and H35B, so the end of the Chothia CDR-H1 loop, when numbered using the Kabat numbering convention shown, varies between H32 and H34 depending on the length of the loop.

[0307] [Table 2]

[0308] Thus, unless otherwise specified, the "CDRs" or "complementary determining regions" of a given antibody or region thereof, e.g., its variable region, or each designated CDR (e.g., CDR-H1, CDR-H2, CDR-H3), will be understood to encompass a (or specific) complementary determining region as defined by any of the schemes described above or other known schemes. For example, a particular CDR (e.g., CDR-H3) may be complementary to a given V H or V L When a variable region amino acid sequence is described as containing the amino acid sequence of a corresponding CDR in the variable region, it is understood that such CDR has the sequence of the corresponding CDR (e.g., CDR-H3) in the variable region as defined by any of the schemes described above or other known schemes. In some embodiments, specific CDR sequences are specified. Although exemplary CDR sequences of the provided antibodies are described using various numbering schemes, it is understood that the provided antibodies can include CDRs described according to any of the other numbering schemes described above or other numbering schemes known to those of skill in the art.

[0309] Similarly, unless otherwise specified, a given antibody or region thereof, e.g., a FR of its variable region or individually designated FR(s) (e.g., FR-H1, FR-H2, FR-H3, FR-H4), will be understood to encompass a (or specific) framework region as defined by any known scheme. In some instances, a scheme for identifying a particular CDR, FR or FRs or CDRs is specified, such as CDRs defined by the Kabat, Chothia, AbM, IMGT, or contact method or other known scheme. In other cases, the specific amino acid sequence of a CDR or FR is given.

[0310] The term "variable region" or "variable domain" refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to an antigen. The variable regions of the heavy and light chains (V, respectively) of a native antibody H and V L ) generally have a similar structure, with each domain containing four conserved framework regions (FR) and three CDRs (see, e.g., Kindt et al., Kuby Immunology, 6th ed., W.H. Freeman and Co., page 91 (2007)). H or V L Each complementary V domain may be sufficient to confer antigen-binding specificity. L or V H To screen a library of domains, we used V from an antibody that binds to an antigen. H or V L Domains can be used to isolate antibodies that bind to specific antigens. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0311] Among the antigen-binding domains contained in CARs, antibody fragments are particularly mentioned. "Antibody fragment" or "antigen-binding fragment" refers to a molecule other than an intact antibody that contains a portion of the intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; heavy chain variable (V); H ) regions, single chain antibody molecules, e.g., scFv, and V H In certain embodiments, the antibody comprises a heavy chain variable (V), such as an scFv. H ) region and / or light chain variable (V L ) region.

[0312] A single domain antibody (sdAb) is an antibody fragment that contains all or part of the heavy chain variable region or all or part of the light chain variable region of an antibody. In certain embodiments, the single domain antibody is a human single domain antibody. In certain embodiments, the single domain antibody is a human single domain antibody. In some embodiments, the CAR comprises an antibody heavy chain domain that specifically binds to BCMA.

[0313] Antibody fragments can be produced by a variety of techniques, including, but not limited to, proteolytic digestion of intact antibodies or production by recombinant host cells. In some embodiments, the antibody is a recombinantly produced fragment, such as a fragment having two or more antibody regions or chains joined by a synthetic linker, e.g., a peptide linker, and / or a fragment containing a non-naturally occurring configuration, such as a fragment that cannot be produced by enzymatic digestion of a naturally occurring intact antibody. In some aspects, the antibody fragment is an scFv.

[0314] In some embodiments, the CAR is an sdFv, nanobody, V H H and V NAR etc., the variable weight (V) of a monoclonal antibody (mAb) or single domain antibody (sdAb) H ) and variable light (V L In some embodiments, the antigen-binding fragment comprises an antigen-binding portion or portions of an antibody molecule, such as a single-chain antibody fragment (scFv) derived from an antibody variable region (V) linked by a flexible linker.

[0315] In some embodiments, the antibody or antigen-binding fragment thereof is a single-chain antibody fragment, such as a single-chain variable fragment (scFv) or a diabody or a single-domain antibody (sdAb). H In some embodiments, the CAR is a single domain antibody comprising only the V domain. In some embodiments, the CAR comprises an sdAb. In some embodiments, the CAR comprises two sdAbs. In some embodiments, each of the two sdAbs is a V HIn some embodiments, each of the two sdAbs binds to a different epitope of BCMA. In some embodiments, each of the two sdAbs binds to the same epitope of BCMA. In some embodiments, the antibody or antigen-binding domain comprises a heavy chain variable (V H ) region and the light chain variable (V L ) region.

[0316] A "humanized" antibody is an antibody in which all or substantially all CDR amino acid residues are derived from non-human CDRs and all or substantially all FR amino acid residues are derived from human FRs. A humanized antibody may comprise at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of a non-human antibody typically refers to a variant of a non-human antibody that has undergone humanization to reduce immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. In some embodiments, some FR residues in a humanized antibody are substituted with corresponding residues from a non-human antibody (e.g., the antibody from which the CDR residues are derived) to, for example, restore or improve antibody specificity or affinity.

[0317] Among the anti-BCMA antibodies included in the provided CARs, murine antibodies are particularly mentioned. A "murine antibody" is an antibody having an amino acid sequence corresponding to the sequence of an antibody produced by a mouse or mouse cell, or a murine antibody repertoire, including a murine antibody library, or other non-murine source that utilizes murine antibody coding sequences.

[0318] Among the anti-BCMA antibodies contained in the provided CARs, human antibodies are particularly preferred. A "human antibody" is an antibody having an amino acid sequence corresponding to that of an antibody produced by a human or human cell, or a non-human source utilizing a human antibody repertoire, including a human antibody library, or other human antibody coding sequence. This term excludes humanized forms of non-human antibodies containing non-human antigen-binding regions, such as those in which all or substantially all CDRs are non-human. This term includes antigen-binding fragments of human antibodies.

[0319] Human antibodies can be prepared by administering immunogens to transgenic animals that have been engineered to produce intact human antibodies or intact antibodies with human variable regions in response to antigen challenge. Such animals typically contain all or part of the human immunoglobulin loci, which replace the endogenous immunoglobulin loci or are extrachromosomally present or randomly integrated into the animal's chromosomes. In such transgenic animals, the endogenous immunoglobulin loci are generally inactivated. Human antibodies can also be derived from human antibody libraries, including phage display and cell-free libraries, containing antibody coding sequences derived from the human repertoire.

[0320] Among the antibodies contained in the provided CAR, particular mention is made of monoclonal antibodies, including monoclonal antibody fragments. As used herein, the term "monoclonal antibody" refers to an antibody obtained from or contained within a substantially homogeneous antibody population, i.e., the individual antibodies constituting the population are identical except for possible variants that contain naturally occurring mutations or arise during the production of the monoclonal antibody preparation, and such variants are generally present in minor amounts. In contrast to polyclonal antibody preparations, which typically contain different antibodies directed against different epitopes, each monoclonal antibody in a monoclonal antibody preparation is directed against a single epitope in an antigen. This term should not be interpreted as requiring the production of antibodies by any particular method. Monoclonal antibodies can be produced by a variety of techniques, including, but not limited to, hybridoma generation, recombinant DNA methods, phage display, and other antibody display methods.

[0321] Thus, in some embodiments, the chimeric antigen receptor comprising a TCR-like CAR comprises an extracellular portion comprising an antibody or antibody fragment. In some embodiments, the antibody or fragment comprises an scFv. In some embodiments, the antibody or antigen-binding fragment is a single-chain antibody fragment, such as a single-chain variable fragment (scFv) or a diabody or single-domain antibody (sdAb). In some embodiments, the antibody or antigen-binding fragment comprises a V H In some embodiments, the antibody or antigen-binding domain is a single domain antibody comprising only the heavy chain variable (V H ) region and the light chain variable (V L ) region.

[0322] In some embodiments, the antibody comprises a heavy chain variable (V H ) region and the light chain variable (V Land antigen-binding fragments, such as scFvs, that include one or more linkers connecting two antibody domains or regions, such as the α- and β-domain domains. The linker is typically a peptide linker, e.g., a flexible and / or soluble peptide linker. Among linkers, linkers rich in glycine and serine and / or, in some cases, threonine are particularly preferred. In some embodiments, the linker further comprises charged residues, such as lysine and / or glutamic acid, which can improve solubility. In some embodiments, the linker further comprises one or more prolines. In some aspects, a glycine and serine (and / or threonine)-rich linker comprises at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of such amino acid(s). In some embodiments, it comprises at least or about 50%, 55%, 60%, 70%, or 75% glycine, serine, and / or threonine. In some embodiments, the linker is composed substantially entirely of glycine, serine, and / or threonine. Linkers are generally between about 5 and about 50 amino acids in length, typically between 10 or about 10 and 30 or about 30, e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30, and in some examples, between 10 and 25 amino acids in length. Exemplary linkers include linkers having various numbers of repeats of the sequence GGGGS (4GS; SEQ ID NO: 26) or GGGS (3GS; SEQ ID NO: 27), e.g., between 2, 3, 4, or 5 repeats of such a sequence. Exemplary linkers include linkers having or consisting of the sequence set forth in SEQ ID NO: 28 (GGGGSGGGGSGGGGS), SEQ ID NO: 29 (GSTSGSGKPGSGEGSTKG), SEQ ID NO: 30 (SRGGGGSGGGGSGGGGSLEMA), or SEQ ID NO: 38 (ASGGGGSGGRASGGGGS). In some embodiments, the linker is or comprises the sequence set forth in SEQ ID NO: 29.

[0323] In some embodiments, the CAR comprises the heavy chain variable (VH ) region and / or light chain variable (V L) region, e.g., the BCMA-binding portion(s) of an antibody molecule, such as an scFv antibody fragment. Chimeric receptors such as CARs generally comprise a portion of an antibody molecule, generally an extracellular antigen-binding domain, such as the variable heavy (VH) chain region and / or variable light (VL) chain region of an antibody, e.g., an scFv antibody fragment. In some embodiments, the provided BCMA-binding CARs contain an antibody, such as an anti-BCMA antibody or antigen-binding fragment thereof, that confers the BCMA-binding properties of the provided CAR. In some embodiments, the antibody or antigen-binding domain can be or can be derived from any of the anti-BCMA antibodies described. See, e.g., Carpenter et al., Clin. Cancer Res., 2013, 19(8):2048-2060; Feng et al., Scand. J. Immunol. (2020) 92:e12910; U.S. Patent No. 9,034,324, U.S. Patent No. 9,765,342; U.S. Patent Publication Nos. US2016 / 0046724, US20170183418; and International Published PCT Application Nos. WO2016090320, WO2016090327, WO2016094304, WO2016014565, WO106014789, WO2010104949, WO2017 / 025038 or WO2017173256, WO2018085690 or WO2021091978. Any of such anti-BCMA antibodies or antigen-binding fragments can be used in the provided CARs. In some embodiments, the anti-BCMA CAR contains one or more single-domain anti-BCMA antibodies. In some embodiments, the one or more single domain anti-BCMA antibodies are derived from an antibody described in WO2017025038 or WO2018028647. In some embodiments, the anti-BCMA CAR contains two single domain anti-BCMA antibodies. In some embodiments, the two single domain anti-BCMA antibodies are derived from one or more antibodies (with or without signal peptides) described in WO2017025038 or WO2018028647.In some embodiments, the BCMA binding domain comprises or consists of A37353-G4S-A37917 (G4S is a linker between the two binding domains), as described in WO2017025038 or WO2018028647, and as provided, for example, in SEQ ID NOs: 300, 301 and 302 of WO2017025038 or WO2018028647. In some embodiments, the anti-BCMA CAR comprises a variable weight (V H ) area and / or variable light (V L In some embodiments, the antigen-binding domain is an scFv containing a variable heavy (V) region. H ) area and / or variable light (V L In some embodiments, the scFv containing the variable heavy (V) region is derived from an antibody described in WO2016090320 or WO2016090327. H ) area and / or variable light (V L In some embodiments, the scFv containing the variable-heavy (V) region is derived from an antibody described in WO2019 / 090003. H ) area and / or variable light (V L In some embodiments, the scFv containing the variable heavy (V) region is derived from an antibody described in WO2016094304 or WO2021091978. H ) area and / or variable light (V L In some embodiments, the scFv containing the variable heavy (V) region is derived from an antibody described in WO2018133877. H ) area and / or variable light (V L ) region is derived from an antibody described in WO2019149269. In some embodiments, the anti-BCMA CAR is any of those described in WO2019173636 or WO2020051374A. In some embodiments, the anti-BCMA CAR is any of those described in WO2018102752. In some embodiments, the anti-BCMA CAR is any of those described in WO2020112796 or WO2021173630.

[0324] In some embodiments, the antibody, e.g., anti-BCMA antibody or antigen-binding fragment, comprises a heavy and / or light chain variable (V) as described. H or V L In some embodiments, the anti-BCMA antibody, e.g., antigen-binding fragment, contains a V domain sequence, VH1, VH2, and / or VH3, as described. H In some embodiments, the anti-BCMA antibody, e.g., antigen-binding fragment, comprises a V domain sequence containing CDR-L1, CDR-L2, and / or CDR-L3 as described. L In some embodiments, the anti-BCMA antibody, e.g., antigen-binding fragment, comprises a V domain sequence or a sufficient antigen-binding portion thereof containing CDR-H1, CDR-H2, and / or CDR-H3 as described. H V containing the region sequences and containing CDR-L1, CDR-L2 and / or CDR-L3 as described L Particularly included are antibodies having a sequence that is at least or about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to such a sequence.

[0325] In some embodiments, the antibody is a V H V such as any of the sequences (e.g., CDR-H1, CDR-H2, CDR-H3 and / or CDR-H4) H and single domain antibodies (sdAbs) that contain only the domain sequence or a sufficient antigen-binding portion thereof.

[0326] In some embodiments, V H The antibodies (e.g., anti-BCMA antibodies) or antigen-binding fragments thereof provided herein that comprise a V region further comprise a light chain or a sufficient antigen-binding portion thereof. For example, in some embodiments, the antibodies or antigen-binding fragments thereof comprise a V region. H Area and V L Area, or V H and VL In such embodiments, the V H The domain sequence is V H In some such embodiments, the antibody is an antigen-binding fragment, such as a Fab or scFv. In some such embodiments, the antibody is a full-length antibody that also contains a constant region.

[0327] In some embodiments, the CAR is an anti-BCMA CAR specific for BCMA, for example, human BCMA. Chimeric antigen receptors containing anti-BCMA antibodies, including mouse anti-human BCMA antibodies and human anti-human BCMA antibodies, and cells expressing such chimeric receptors have been previously described. See Carpenter et al., Clin Cancer Res., 2013, 19(8):2048-2060, US9,765,342, WO2016 / 090320, WO2016090327, WO2010104949A2, WO2016 / 0046724, WO2016 / 014789, WO2016 / 094304, WO2017 / 025038 and WO2017173256.

[0328] In some embodiments, the anti-BCMA CAR comprises a variable-heavy (V) antibody derived from an antibody described in WO2016094304 or WO2021091978. H ) area and / or variable light (V L In some embodiments, the antigen-binding domain is an antibody fragment containing a variable heavy (VH) region and a variable light (VL) region. In some embodiments, the anti-BCMA CAR contains an antigen-binding domain such as an scFv containing a variable heavy (VH) region and a variable light (VL) region from an antibody described in WO2016 / 090320 or WO2016090327. H ) area and / or variable light (V L ) region.

[0329] In some embodiments, the antigen-binding domain comprises a variable heavy chain (V H ) region and the variable light chain (VL In some embodiments, the antibody fragment contains a V H The region may be any of the V sequences set forth in any of SEQ ID NOs: 18, 20, 22, 24, 32, 34, 36, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 145, 147, 149, and 151. H is or comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of the V L The region may be any of the V sequences set forth in any of SEQ ID NOs: 19, 21, 23, 25, 33, 35, 37, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 146, 148, 150, and 152. L It is or comprises an amino acid sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of the region.

[0330] In some embodiments, the antigen binding domain, such as an scFv, comprises the V H and V as set forth in SEQ ID NO: 19 L In some embodiments, the antigen binding domain, such as an scFv, contains the V H and V as set forth in SEQ ID NO: 21 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 22. H and V as set forth in SEQ ID NO: 23 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 24. H and V set forth in SEQ ID NO: 25 L In some embodiments, the antigen binding domain, such as an scFv, comprises the V H and V set forth in SEQ ID NO: 33 LIn some embodiments, the antigen binding domain, such as an scFv, comprises the V H and V set forth in SEQ ID NO: 35 L In some embodiments, the antigen binding domain, such as an scFv, comprises the V H and V set forth in SEQ ID NO: 37 L In some embodiments, the antigen binding domain, such as an scFv, comprises the V set forth in SEQ ID NO: 41. H and V set forth in SEQ ID NO: 42 L In some embodiments, the antigen binding domain, such as an scFv, comprises the V H and V set forth in SEQ ID NO: 44 L In some embodiments, the antigen binding domain, such as an scFv, comprises the V H and V set forth in SEQ ID NO: 46 L In some embodiments, the antigen binding domain, such as an scFv, comprises the V set forth in SEQ ID NO: 47. H and V set forth in SEQ ID NO: 48 L In some embodiments, the antigen binding domain, such as an scFv, comprises the V set forth in SEQ ID NO:49. H and V as set forth in SEQ ID NO: 50 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 51. H and V set forth in SEQ ID NO: 52 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 53. H and V set forth in SEQ ID NO: 54 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 55. H and V set forth in SEQ ID NO: 56 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 57. H and V set forth in SEQ ID NO: 58 LIn some embodiments, the antigen binding domain, such as an scFv, comprises the V set forth in SEQ ID NO: 59. H and V set forth in SEQ ID NO: 60 L In some embodiments, the antigen binding domain, such as an scFv, comprises the V H and V set forth in SEQ ID NO: 62 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 63. H and V set forth in SEQ ID NO: 64 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 65. H and V set forth in SEQ ID NO: 66 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 67. H and V set forth in SEQ ID NO: 68 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 69. H and V set forth in SEQ ID NO: 70 L In some embodiments, the antigen binding domain, such as an scFv, contains the V H and V set forth in SEQ ID NO: 72 L In some embodiments, the antigen binding domain, such as an scFv, contains the V set forth in SEQ ID NO: 73. H and V set forth in SEQ ID NO: 74 L In some embodiments, the antigen binding domain, such as an scFv, contains the V H and V set forth in SEQ ID NO: 76 L In some embodiments, the antigen binding domain, such as an scFv, contains the V H and V set forth in SEQ ID NO: 146 L In some embodiments, the antigen binding domain, such as an scFv, contains the V H and V set forth in SEQ ID NO: 148 LIn some embodiments, the antigen binding domain, such as an scFv, contains the V H and V set forth in SEQ ID NO: 150 L In some embodiments, the antigen binding domain, such as an scFv, contains the V H and V set forth in SEQ ID NO: 152 L In some embodiments, V H or V L is the aforementioned V H or V L In some embodiments, the V has a sequence of amino acids that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to any of the V sequences and retains binding to BCMA. H The region is V L In some embodiments, the V H The region is V L In some embodiments, the variable heavy chain and the variable light chain are connected by a linker. In some embodiments, the linker is set forth in SEQ ID NO: 28, 29, 30, or 38.

[0331] Among the anti-BCMA CARs provided are antibodies or antigen-binding fragments comprising a V that comprises the sequence set forth in SEQ ID NO: 18, or an amino acid sequence having at least or about 90%, 91% or about 91%, 92% or about 92%, 93% or about 93%, 94% or about 94%, 95% or about 95%, 96% or about 96%, 97% or about 97%, 98% or about 98%, or 99% or about 99% identity to SEQ ID NO: 18. Hand a V comprising a sequence set forth in SEQ ID NO:19 or an amino acid sequence having at least or about 90%, 91% or about 91%, 92% or about 92%, 93% or about 93%, 94% or about 94%, 95% or about 95%, 96% or about 96%, 97% or about 97%, 98% or about 98%, or 99% or about 99% identity to SEQ ID NO:19. L In some embodiments, the antibody or antigen-binding fragment of the CAR provided has a V domain having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 189, 190, and 191, respectively. H and a V region having CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 192, 193, and 194, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V domain having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 195, 196, and 197, respectively. H and a V region having CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 198, 199, and 200, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V domain having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 201, 20...

Claims

1. 1. A method of treating a subject with multiple myeloma (MM), comprising: a dose of genetically engineered T cells, the T cells expressing a chimeric antigen receptor (CAR) that binds to human BCMA; (a) the subject is (i) having a serum soluble B-cell maturation antigen (sBCMA) level of less than about 600 ng / mL; and / or (ii) Absence of IgG heavy chain disease (HCD) and determining (b) administering to the subject a dose of genetically engineered T cells, including T cells expressing a chimeric antigen receptor (CAR) that binds to human BCMA; and A pharmaceutical agent for use in a method comprising:

2. 1. A method of treating a subject with multiple myeloma (MM), comprising administering a dose of genetically engineered T cells comprising T cells expressing a chimeric antigen receptor (CAR) that binds to human B-cell maturation antigen (BCMA) to a subject with multiple myeloma (MM), the method comprising administering a dose of genetically engineered T cells comprising T cells expressing a chimeric antigen receptor (CAR) that binds to human B-cell maturation antigen (BCMA) to a subject with multiple myeloma (MM). (i) having a serum soluble B-cell maturation antigen (sBCMA) level of less than about 600 ng / mL; and / or (ii) Absence of IgG heavy chain disease (HCD) A pharmaceutical agent for use in a method comprising administering to a subject a predetermined amount of a compound.

3. 1. A method of treating a subject with multiple myeloma (MM), comprising: a dose of genetically engineered T cells, the T cells expressing a chimeric antigen receptor (CAR) that binds to human BCMA; (a) for treatment with a dose of genetically engineered T cells, including T cells expressing a chimeric antigen receptor (CAR) that binds to human BCMA; (i) having a serum soluble B-cell maturation antigen (sBCMA) level of less than about 600 ng / mL; and / or (ii) Absence of IgG heavy chain disease (HCD) selecting a target whose value is predetermined; (b) administering a dose of the genetically engineered T cells to the subject; A pharmaceutical agent for use in a method comprising:

4. The method of claim 1, wherein the subject is predicted to achieve a complete response (CR) or a stringent complete response (sCR) if the subject has (i) and / or (ii).

5. 1. A method of treating a subject with multiple myeloma (MM), comprising: a dose of genetically engineered T cells, the T cells expressing a chimeric antigen receptor (CAR) that binds to human BCMA; (a) the subject is (i) having a serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL; and / or (ii) determining the presence of IgG heavy chain disease (HCD); (b) debulking the MM; (c) administering to the subject a dose of genetically engineered T cells, including T cells expressing a chimeric antigen receptor (CAR) that binds to human BCMA; and A pharmaceutical agent for use in a method comprising:

6. The method of claim 5, wherein the multiple myeloma (MM) was debulked between (1) the subject's determination that they have (i) and / or (ii) and (2) the subject's administration of a dose of the engineered T cells.

7. 6. The method of claim 5, wherein if the subject is determined to have (i) and / or (ii), the subject is selected for debulking of MM prior to administration to the subject of a dose of genetically engineered T cells comprising T cells expressing a chimeric antigen receptor (CAR) that binds to human BCMA.

8. If the subject has (i) and / or (ii), the subject is predicted not to achieve a complete response (CR) or stringent complete response (sCR); The method of claim 5, wherein the treatment comprises administering to the subject a dose of genetically engineered T cells.

9. 1. A method of treating a subject with multiple myeloma (MM), comprising: a dose of genetically engineered T cells, the T cells expressing a chimeric antigen receptor (CAR) that binds to human BCMA; (a) determining that the subject has a first serum soluble B-cell maturation antigen (sBCMA) level greater than about 600 ng / mL; (b) debulking the MM; (c) determining that the subject has a post-debulking serum sBCMA level less than about 600 ng / mL; (d) administering to the subject a dose of genetically engineered T cells, including T cells expressing a chimeric antigen receptor (CAR) that binds to human BCMA; and A pharmaceutical agent for use in a method comprising:

10. The method is (i) a serum sBCMA level or a post-debulking serum sBCMA level greater than or less than about 600 ng / mL; and / or (ii) the presence or absence of IgG HCD The method of claim 9, further comprising determining:

11. The method includes determining that the subject has a serum sBCMA level greater than or less than about 590 ng / ml, about 580 ng / ml, about 570 ng / ml, about 566 ng / ml, about 560 ng / ml, about 550 ng / ml, about 540 ng / ml, about 530 ng / ml, about 520 ng / ml, about 510 ng / ml, or about 500 ng / ml; and / or 10. The method of any one of claims 1 to 9, wherein the subject is determined to have a serum sBCMA level greater than or less than about 590ng / ml, about 580ng / ml, about 570ng / ml, about 566ng / ml, about 560ng / ml, about 550ng / ml, about 540ng / ml, about 530ng / ml, about 520ng / ml, about 510ng / ml, or about 500ng / ml.

12. The method of any one of claims 1 to 9, wherein determining that the subject has the presence of IgG HCD comprises detecting IgG in the serum and / or urine of the subject.

13. 10. The method of any one of claims 1 to 9, wherein the determination of the serum sBCMA level is performed (i) about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week prior to administration of the dose of engineered T cells to the subject, or (ii) about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week prior to obtaining the dose of engineered T cells from the subject.

14. 10. The method of any one of claims 1-9, wherein the determination that the subject has or does not have the presence of IgG HCD is performed about 3 months, about 2 months, about 1 month, about 3 weeks, about 2 weeks, or about 1 week prior to (i) administering the dose of engineered T cells to the subject, or (ii) obtaining the dose of engineered T cells from the subject.

15. The method of any one of claims 1 to 9, wherein the subject achieves a complete response (CR) or stringent complete response (sCR) following administration of a dose of the genetically engineered T cells to the subject.

16. The method of claim 5 , wherein debulking comprises administering chemotherapy, radiation, and / or an immunomodulatory agent to the subject.

17. 17. The method of claim 16, wherein the chemotherapy comprises melphalan, doxorubicin, or cyclophosphamide chemotherapy.

18. 17. The method of claim 16, wherein the immunomodulatory agent is thalidomide, lenalidomide, or pomalidomide.

19. The method of claim 16 , wherein the immunomodulatory agent is a checkpoint inhibitor.

20. 10. The agent of any one of claims 5, 7, and 9, wherein the debulking is performed within about 3 months, within about 2 months, within about 1 month, within about 3 weeks, within about 2 weeks, or within about 1 week prior to administration of the dose of engineered T cells to the subject.

21. Prior to administration of a dose of engineered T cells to the subject, the subject administers fludarabine to a patient having a body surface area of ​​1 m ２ and / or administering cyclophosphamide at or about 20-40 mg / m² of body surface area to the subject daily for 2-4 days. ２ 10. The method of claim 1, wherein the patient is undergoing lymphodepletion therapy comprising administering at or about 200-400 mg per day for 2-4 days.

22. Prior to administration of a dose of engineered T cells to the subject, the subject administers fludarabine to a patient having a body surface area of ​​1 m ２ Cyclophosphamide was administered daily at 30 mg or approximately 30 mg per m² of body surface area. ２ 10. The method of any one of claims 1 to 9, wherein the patient is undergoing lymphodepletion therapy comprising administering at or about 300 mg per day for three days.

23. 22. The method of claim 21, wherein debulking is performed before lymphodepletion therapy or wherein debulking is performed after lymphodepletion therapy.

24. The method of any one of claims 1 to 9, wherein the MM is high-risk MM and / or relapsed and / or refractory (r / r) MM.

25. The method according to any one of claims 1 to 9, wherein the subject is 18 years of age or older.

26. The subject has previously received 3 or more prior lines of therapy for MM, where: Three or more prior lines of therapy each included two consecutive cycles unless progressive disease (PD; progression within 60 days after the last dose) was the best response to that line of therapy; three or more prior lines of therapy include a proteasome inhibitor, an immunomodulatory agent, and an anti-CD38 antibody; and / or The subject is refractory to the last of three or more prior lines of therapy. The drug according to any one of claims 1 to 9.

27. The method of any one of claims 1 to 9, wherein the subject has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

28. the subject has measurable disease upon administration of the dose of engineered T cells, wherein the measurable disease is (i) serum M protein 1.0 g / dL or more; (ii) urinary M protein ≥ 200 mg / 24 hours; and / or (iii) if the serum free light chain (FLC) ratio is abnormal, a diseased serum FLC level of 10 mg / dL or more; The drug according to any one of claims 1 to 9, comprising:

29. The target is, (i) central nervous system (CNS) lesions; (ii) a history or presence of a clinically relevant CNS condition; and / or (iii) active plasma cell leukemia (PCL) or a history of PCL The drug according to any one of claims 1 to 9, which does not have

30. wherein the CAR comprises an extracellular antigen-binding domain that binds to BCMA, a transmembrane domain, and an intracellular signaling region; (i) the extracellular antigen-binding domain is a variable heavy chain (V Ｈ ) region, and optionally a variable light chain (V Ｌ ) area; (ii) V Ｈ the regions comprising CDR-H1, CDR-H2 and CDR-H3 comprising the amino acid sequences set forth in SEQ ID NOs: 189, 190 and 191, respectively; Ｌ the regions comprise CDR-L1, CDR-L2 and CDR-L3 comprising the amino acid sequences set forth in SEQ ID NOs: 192, 193 and 194, respectively; or V Ｈ the regions comprising CDR-H1, CDR-H2 and CDR-H3 comprising the amino acid sequences set forth in SEQ ID NOs: 173, 174 and 175, respectively; Ｌ the regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences set forth in SEQ ID NOs: 183, 184, and 185, respectively; (iii) V Ｈ The region comprises the amino acid sequence set forth in SEQ ID NO: 18, Ｌ the region comprises the amino acid sequence set forth in SEQ ID NO: 19; or V Ｈ The region comprises the amino acid sequence set forth in SEQ ID NO: 24, Ｌ the region comprises the amino acid sequence set forth in SEQ ID NO: 25; and / or (iv) the extracellular antigen-binding domain is a single-chain variable fragment (scFv), and the scFv comprises the amino acid sequence set forth in SEQ ID NO: 213 or SEQ ID NO: 188; The drug according to any one of claims 1 to 9.

31. wherein the CAR comprises an extracellular antigen-binding domain that binds to BCMA, a transmembrane domain, and an intracellular signaling region; (i) the intracellular signaling region further comprises a costimulatory signaling domain, wherein the costimulatory signaling domain comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof, and wherein the costimulatory signaling domain is between the transmembrane domain and the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain; and / or (ii) the transmembrane domain is or comprises a transmembrane domain derived from CD28 or CD8; The drug of claim 30.

32. The agent of claim 31, wherein the CD28 or CD8 is human CD28 or CD8.

33. The agent of any one of claims 1 to 9, wherein the CAR further comprises an extracellular spacer between the antigen-binding domain and the transmembrane domain, wherein the spacer is derived from CD8, and / or the transmembrane domain and the spacer are derived from CD8.

34. The agent of claim 33, wherein the extracellular spacer is a CD8α hinge.

35. The agent of any one of claims 1 to 9, wherein the CAR comprises the amino acid sequence set forth in SEQ ID NO: 116 or SEQ ID NO: 124, and / or the CAR is encoded by the polynucleotide sequence set forth in SEQ ID NO:

214.

36. The dose of genetically engineered T cells was determined by the following: idecbutagen biculeucel cells; bb21217 cells; orbacabutagen autreucel cells; CT103A cells; siltacbutagen autreucel cells; KITE585 cells; CT053 cells; BCMA-CS1 The agent of any one of claims 1 to 9, comprising cCAR (BC1cCAR) cells; P-BCMA-101 cells; P-BCMA-ALLO1 cells; C-CAR088 cells; Descartes-08 cells; PBCAR269A cells; ALLO-715 cells; PHE885 cells; AUTO8 cells; CTX120 cells; CB-011 cells; ALLO-605 (TuboCAR / MM) cells; pCDCAR1 (TriCAR-Z136) cells, or GC012F cells.

37. The dose of engineered T cells is ＋ The agent according to any one of claims 1 to 9, comprising CAR-expressing T cells.

38. The dose of engineered T cells is ＋ T cells and CD8 ＋ Combination of T cells and / or CD4 ＋ CAR-expressing T cells and CD8 ＋ a combination of CAR-expressing T cells, wherein CD4 ＋ CAR-expressing T cells vs. CD8 ＋ CAR-expressing T cells and / or CD4 ＋ T cells vs. CD8 ＋ The method of any one of claims 1 to 9, wherein the ratio of T cells is at or about 1:1 or between 1:3 or about 1:3 and 3:1 or about 3:

1.

39. The dose of engineered T cells is naive-like T cells and / or central memory T cells at a percentage greater than or equal to about 60% of the total engineered T cells in the dose; naive-like T cells and / or central memory T cells at a percentage greater than or equal to 40% of the total CD4+ engineered T cells in the dose; or comprising naive-like T cells and / or central memory T cells at a percentage greater than or equal to about 40% of the total CD8+ engineered T cells in the dose; and / or naive-like T cells that are CCR7+CD45RA+, CD27+CCR7+, or CD62L-CCR7+; The drug according to any one of claims 1 to 9.

40. The dose of engineered T cells is approximately 0.5 x 10 ６ ~Approx. 600×10 ６ CAR-positive T cells, approximately 100 x 10 ６ ~Approx. 600×10 ６ CAR-positive T cells, or approximately 150 x 10 ６ ~Approx. 450×10 ６ The agent according to any one of claims 1 to 9, comprising CAR-positive T cells.

41. The dose of genetically engineered T cells was approximately 150 x 10 ６ , 300 x 10 ６ , or about 450 × 10 ６ The agent according to any one of claims 1 to 9, comprising CAR-positive T cells.

42. The dose of engineered T cells is approximately 0.5 x 10 ６ ~About 10×10 ６ The agent according to any one of claims 1 to 9, comprising CAR-positive T cells.

43. The method of any one of claims 1 to 9, wherein the dose of genetically engineered T cells is obtained from a subject.

44. The method of any one of claims 1 to 9, wherein the cells of the dose of genetically engineered T cells are autologous to the subject.

45. The method of any one of claims 1 to 9, wherein the cells of the dose of genetically engineered T cells are allogeneic to the subject.

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