Bcma-directed chimeric antigen receptor t cell compositions and methods and uses thereof

Abstract

Provided in some aspects are compositions of cells for treating subjects with disease and conditions such as multiple myeloma (MM), and related methods, compositions, uses and articles of manufacture. In some embodiments, the disease or condition is a relapsed or refractory multiple myeloma (r / r MM). The cells generally express recombinant receptors such as chimeric antigen receptors (CARs) for targeting an antigen, such as BCMA, on cells of the myeloma.

Description

BCMA-DIRECTED CHIMERIC ANTIGEN RECEPTOR T CELL COMPOSITIONS AND METHODS AND USES THEREOF Cross-Reference to Related Applications

[0001] This application claims priority from U.S. provisional application No.62 / 975,731 filed February 12, 2020, entitled “BCMA-DIRECTED CHIMERIC ANTIGEN RECEPTOR T CELL COMPOSITIONS AND METHODS AND USES THEREOF,” the contents of which are incorporated by reference in their entirety. Incorporation by Reference of Sequence Listing

[0002] The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled 735042023540SEQLIST.txt, created February 10, 2021, which is 184 kilobytes in size. The information in the electronic format of the Sequence Listing is incorporated by reference in its entirety. Field

[0003] The present disclosure relates in some aspects to adoptive cell therapy involving the administration of compositions of cells for treating subjects with disease and conditions such as multiple myeloma (MM), and related methods, compositions, uses and articles of manufacture. Background

[0004] Various immunotherapy and / or cell therapy methods are available for treating diseases and conditions. For example, adoptive cell therapies (including those involving the administration of cells expressing chimeric receptors specific for a disease or disorder of interest, such as chimeric antigen receptors (CARs) and / or other recombinant antigen receptors, as well as other adoptive immune cell and adoptive T cell therapies) can be beneficial in the treatment of cancer or other diseases or disorders. Improved approaches are needed. Provided are methods, uses and articles of manufacture that meet such needs. Summary

[0005] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method comprising administering to a subject having or suspected of having a MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen(BCMA), wherein: the composition comprises CD4+T cells expressing the CAR and CD8+T cells expressing the CAR; the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 200 × 106CAR-expressing T cells, inclusive; and at least or at least about 80% of the cells in the composition are CD3+cells.

[0006] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method comprising administering to a subject having or suspected of having a MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein: the composition comprises CD4+T cells expressing the CAR and CD8+T cells expressing the CAR at a ratio between about 1:2.5 and about 5:1; the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 200 × 106CAR-expressing T cells, inclusive; and at least or at least about 90% of the cells in the composition are CD3+cells.

[0007] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method comprising administering to a subject having or suspected of having a MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein: the composition comprises CD8+T cells expressing the CAR and CD4+T cells expressing the CAR; the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 200 × 106CAR-expressing T cells, inclusive; at least or at least about 80% of the cells in the composition are CD3+cells; and at least or at least about 80% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype.

[0008] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method comprising administering to a subject having or suspected of having a MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein: the composition comprises CD8+T cells expressing the CAR and CD4+T cells expressing the CAR; the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 200 × 106CAR-expressing T cells, inclusive; at least or at least about 80% of the cells in the composition are CD3+cells; and at least or at least about 50% of the CD4+CAR+T cells in the composition are CD27+CCR7+and / or at least or at least about 50% of the CD8+CAR+T cells in the composition are CD27+CCR7+.

[0009] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method comprising administering to a subject having or suspected of having a MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein: the composition comprises CD8+T cells expressing the CAR and CD4+T cells expressing the CAR; the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 200 × 106CAR-expressing T cells, inclusive; at least or at least about 80% of the cells in thecomposition are CD3+cells; and the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is less than or less than about 0.9.

[0010] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method comprising administering to a subject having or suspected of having a MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein: the composition comprises CD8+T cells expressing the CAR and CD4+T cells expressing the CAR; the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 200 × 106CAR-expressing T cells, inclusive; at least or at least about 80% of the cells in the composition are CD3+cells; and the integrated vector copy number (iVCN) of the CAR+T cells in the composition, on average, is between or between about 0.4 copies per diploid genome and 2.0 copies per diploid genome, inclusive.

[0011] In some of any embodiments, the composition comprises between at or about 50 × 106CAR- expressing T cells and at or about 200 × 106CAR-expressing T cells, inclusive. In some of any embodiments, the composition comprises between at or about 70 × 106CAR-expressing T cells and at or about 200 × 106CAR-expressing T cells, inclusive. In some of any embodiments, the composition comprises between at or about 80 × 106CAR-expressing T cells and at or about 200 × 106CAR- expressing T cells, inclusive. In some of any embodiments, the composition comprises between at or about 80 × 106CAR-expressing T cells and at or about 160 × 106CAR-expressing T cells, inclusive.

[0012] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method including administering to a subject having or suspected of having a MM a composition including engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein the composition includes CD4+T cells expressing the CAR and CD8+T cells expressing the CAR; the composition includes between at or about 5 × 106CAR-expressing T cells and at or about 40 × 106CAR-expressing T cells, inclusive; and at least or at least about 80% of the cells in the composition are CD3+cells.

[0013] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method including administering to a subject having or suspected of having a MM a composition including engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein the composition includes CD4+T cells expressing the CAR and CD8+T cells expressing the CAR at a ratio between about 1:2.5 and about 5:1; the composition includes between at or about 5 × 106CAR-expressing T cells and at or about 80 × 106CAR-expressing T cells, inclusive; at least or at least about 90% of the cells in the composition are CD3+cells.

[0014] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method including administering to a subject having or suspected of having a MM a composition including engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen(BCMA), wherein the composition includes CD4+T cells expressing the CAR and CD8+T cells expressing the CAR; the composition includes between at or about 5 × 106CAR-expressing T cells and at or about 10 × 106CAR-expressing T cells, inclusive; and at least or at least about 80% of the cells in the composition are CD3+cells.

[0015] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method including administering to a subject having or suspected of having a MM a composition including engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein the composition includes CD8+T cells expressing the CAR and CD4+T cells expressing the CAR; the composition includes between at or about 5 × 106CAR-expressing T cells and at or about 80 × 106CAR-expressing T cells, inclusive; at least or at least about 80% of the cells in the composition are CD3+cells; and at least or at least about 80% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype.

[0016] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method including administering to a subject having or suspected of having a MM a composition including engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein the composition includes CD8+T cells expressing the CAR and CD4+T cells expressing the CAR; the composition includes between at or about 5 × 106CAR-expressing T cells and at or about 100 × 106CAR-expressing T cells, inclusive; at least or at least about 80% of the cells in the composition are CD3+cells; and at least or at least about 50% of the CD4+CAR+T cells in the composition are CD27+CCR7+and / or at least or at least about 50% of the CD8+CAR+T cells in the composition are CD27+CCR7+.

[0017] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method including administering to a subject having or suspected of having a MM a composition including engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein the composition includes CD8+T cells expressing the CAR and CD4+T cells expressing the CAR; the composition includes between at or about 5 × 106CAR-expressing T cells and at or about 20 × 106CAR-expressing T cells, inclusive; at least or at least about 80% of the cells in the composition are CD3+cells; and at least or at least about 80% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype.

[0018] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method including administering to a subject having or suspected of having a MM a composition including engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein the composition includes CD8+T cells expressing the CAR and CD4+T cells expressing the CAR; the composition includes between at or about 5 × 106CAR-expressing T cells and at or about 80 × 106CAR-expressing T cells, inclusive; at least or at least about 80% of the cells in thecomposition are CD3+cells; and the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is less than or less than about 0.9.

[0019] In one aspect, provided herein is a method of treating a multiple myeloma (MM), the method including administering to a subject having or suspected of having a MM a composition including engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein the composition includes CD8+T cells expressing the CAR and CD4+T cells expressing the CAR; the composition includes between at or about 5 × 106CAR-expressing T cells and at or about 80 × 106CAR-expressing T cells, inclusive; at least or at least about 80% of the cells in the composition are CD3+cells; and the integrated vector copy number (iVCN) of the CAR+T cells in the composition, on average, is between or between about 0.4 copies per diploid genome and 2.0 copies per diploid genome, inclusive.

[0020] In some of any embodiments, the composition comprises CD4+T cells expressing the CAR and CD8+T cells expressing the CAR at a ratio between about 1:2.5 and about 5:1. In some of any embodiments, the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 80 × 106CAR-expressing T cells, inclusive. In some of any embodiments, the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 40 × 106CAR-expressing T cells, inclusive. In some of any embodiments, the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 20 × 106CAR-expressing T cells, inclusive.

[0021] In some of any embodiments, the composition may include CD4+T cells expressing the CAR and CD8+T cells expressing the CAR at a ratio between about 1:2 and about 4:1, between about 1:1.5 and about 2:1, or at or at about 1:1. In some of any embodiments, the composition may include CD4+T cells expressing the CAR and CD8+T cells expressing the CAR at a ratio between about 5:1 and about 2:1, between about 4:1 and about 2:1, between about 3:1 and about 2:1, at or at about 5:1, at or at about 4:1, at or at about 3:1, or at or at about 2:1. In some of any embodiments, the composition may include between at or about 5 × 106CAR-expressing T cells and at or about 10 × 106CAR-expressing T cells, inclusive. In some of any embodiments, the composition may include between at or about 10 × 106CAR-expressing T cells and at or about 20 × 106CAR-expressing T cells, inclusive. In some of any embodiments, the composition may include at or about 20 × 106CAR-expressing T cells. In some of any embodiments, the composition may include or about 30 × 106CAR-expressing T cells. In some of any embodiments, the composition may include at or about 40 × 106CAR-expressing T cells.In some of any embodiments, the composition may include at or about 10 × 106CAR-expressing T cells. In some of any embodiments, the composition may include at or about 60 × 106CAR-expressing T cells. In some of any embodiments, the composition may include at or about 80 × 106CAR-expressing T cells. In some of any embodiments, the composition may include at or about 160 × 106CAR-expressing T cells.

[0022] In some of any embodiments, at least or at least about 90% of the cells in the composition are CD3+cells.

[0023] In some of any embodiments, at least or at least about 91%, at least or at least about 92%, at least or at least about 93%, at least or at least about 94%, at least or at least about 95%, or at least or at least about 96% of the cells in the composition are CD3+cells. In some of any embodiments, between at or about 2% and at or about 30% of the CAR+T cells in the composition express a marker of apoptosis, optionally Annexin V or active Caspase 3. In some of any embodiments, between at or about 5% and at or about 10% of the CAR+T cells in the composition express a marker of apoptosis, optionally Annexin V or active Caspase 3. In some of any embodiments, between at or about 10% and at or about 15% of the CAR+T cells in the composition express a marker of apoptosis, optionally Annexin V or active Caspase 3. In some of any embodiments, between at or about 15% and at or about 20% of the CAR+T cells in the composition express a marker of apoptosis, optionally Annexin V or active Caspase 3. In some of any embodiments, between at or about 20% and at or about 30% of the CAR+T cells in the composition express a marker of apoptosis, optionally Annexin V or active Caspase 3. In some of any embodiments, at or about 5%, at or about 10%, at or about 15%, at or about 20%, at or about 25%, or at or about 30% of the CAR+T cells in the composition express a marker of apoptosis, optionally Annexin V or active Caspase 3. In some embodiments, the marker of apoptosis is Annexin V. In some embodiments, the marker of apoptosis is active Caspase 3.

[0024] In some of any embodiments, at least or at least about 80% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype.

[0025] In some of any embodiments, between at or about 80% and at or about 85% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype. In some of any embodiments, between at or about 85% and at or about 90% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype. In some of any embodiments, between at or about 90% and at or about 95% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype. In some of any embodiments, between at or about 95% and at or about 99% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype. In some of any embodiments, at or about 85%, at or about 90%, at or about 95%, or at or about 99% of the CAR+T cells in the composition are of a naïve- like or central memory phenotype.

[0026] In some of any embodiments, the at least or at least about 80% of the CAR+T cells in the composition that are of a naïve-like or central memory phenotype are surface positive for a marker expressed on naïve-like or central memory T cells. In some of any embodiments, the marker expressed on naïve-like or central memory T cell is selected from the group consisting of CD45RA, CD27, CD28, and CCR7.

[0027] In some of any embodiments, the at least or at least about 80% of the CAR+T cells in the composition that are of a naïve-like or central memory phenotype have a phenotype selected from CCR7+CD45RA+, CD27+CCR7+, or CD62L-CCR7+. In some of any embodiments, between at or about 80% and at or about 85%, between at or about 85% and at or about 90%, between at or about 90% and at or about 95%, between at or about 95% and at or about 99% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype selected from CCR7+CD45RA+, CD27+CCR7+, or CD62L- CCR7+. In some of any embodiments, at or about 80%, at or about 85%, at or about 90%, at or about 95%, or at or about 99% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype selected from CCR7+CD45RA+, CD27+CCR7+, or CD62L-CCR7+. In some of any embodiments, at or about 80%, at or about 85%, at or about 90%, at or about 95%, or at or about 99% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

[0028] In some of any embodiments, at least or at least about 50% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 60% of the CD4+CAR+T cells in the composition are of a naïve –like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 70% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 80% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 85% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-.

[0029] In some of any embodiments, at least or at least about 50% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+. In some of any embodiments, at least or at least about 60% of the CD4+CAR+T cells in the composition are of a naïve- like or central memory phenotype that is CD27+CCR7+. In some of any embodiments, at least or at least about 70% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+. In some of any embodiments, at least or at least about 80% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+. In some of any embodiments, at least or at least about 85% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

[0030] In some of any embodiments, at least or at least about 50% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 60% of the CD8+CAR+T cellsin the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 70% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 80% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 85% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-.

[0031] In some of any embodiments, at least or at least about 50% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+. In some of any embodiments, at least or at least about 60% of the CD8+CAR+T cells in the composition are of a naïve- like or central memory phenotype that is CD27+CCR7+. In some of any embodiments, at least or at least about 70% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+. In some of any embodiments, at least or at least about 80% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+. In some of any embodiments, at least or at least about 85% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

[0032] In some of any embodiments, at least or at least about 80% of the CAR+T cells in the composition are surface positive for a marker expressed on naïve-like or central memory T cells. In some of any embodiments, the marker expressed on naïve-like or central memory T cell is selected from the group consisting of CD45RA, CD27, CD28, and CCR7. In some of any embodiments, at least or at least about 80% of the CAR+T cells in the composition are CCR7+CD45RA+, CD27+CCR7+, and / or CD62L- CCR7+. In some of any embodiments, between at or about 80% and at or about 85%, between at or about 85% and at or about 90%, between at or about 90% and at or about 95%, between at or about 95% and at or about 99% of the CAR+T cells in the composition are CCR7+CD45RA+, CD27+CCR7+, and / or CD62L-CCR7+. In some of any embodiments, at or about 80%, at or about 85%, at or about 90%, at or about 95%, or at or about 99% of the CAR+T cells in the composition are CCR7+CD45RA+, CD27+CCR7+, and / or CD62L-CCR7+. In some of any embodiments, at or about 80%, at or about 85%, at or about 90%, at or about 95%, or at or about 99% of the CAR+T cells in the composition are CD27+CCR7+. In some of any embodiments, at least or at least about 50% of the CD4+CAR+T cells in the composition are CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 60% of the CD4+CAR+T cells in the composition are CCR7+CD45RA+or CCR7+CD45RA-.

[0033] In some of any embodiments, at least or at least about 70% of the CD4+CAR+T cells in the composition are CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 80% of the CD4+CAR+T cells in the composition are CCR7+CD45RA+or CCR7+CD45RA-. Insome of any embodiments, at least or at least about 85% of the CD4+CAR+T cells in the composition are CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 50% of the CD4+CAR+T cells in the composition are CD27+CCR7+. In some of any embodiments, at least or at least about 60% of the CD4+CAR+T cells in the composition are CD27+CCR7+. In some of any embodiments, at least or at least about 70% of the CD4+CAR+T cells in the composition are CD27+CCR7+. In some of any embodiments, at least or at least about 80% of the CD4+CAR+T cells in the composition are CD27+CCR7+. In some of any embodiments, at least or at least about 85% of the CD4+CAR+T cells in the composition are CD27+CCR7+. In some of any embodiments, at least or at least about 50% of the CD8+CAR+T cells in the composition are CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 60% of the CD8+CAR+T cells in the composition are CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 70% of the CD8+CAR+T cells in the composition are CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 80% of the CD8+CAR+T cells in the composition are CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 85% of the CD8+CAR+T cells in the composition are CCR7+CD45RA+or CCR7+CD45RA-. In some of any embodiments, at least or at least about 50% of the CD8+CAR+T cells in the composition are CD27+CCR7+. In some of any embodiments, at least or at least about 60% of the CD8+CAR+T cells in the composition are CD27+CCR7+. In some of any embodiments, at least or at least about 70% of the CD8+CAR+T cells in the composition are CD27+CCR7+. In some of any embodiments, at least or at least about 80% of the CD8+CAR+T cells in the composition are CD27+CCR7+. In some of any embodiments, at least or at least about 85% of the CD8+CAR+T cells in the composition are CD27+CCR7+.

[0034] In some of any embodiments, the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is less than or less than about 0.9.

[0035] In some of any embodiments, the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is between at or about 0.9 and at or about 0.8. In some of any embodiments, the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is less than or less than about 0.8. In some of any embodiments, the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is between at or about 0.8 and at or about 0.7. In some of any embodiments, the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is between at or about 0.7 and at or about 0.6. In some of any embodiments, the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is between at or about 0.6 and at or about 0.5. In some of any embodiments, the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is between at or about 0.5 and at or about 0.4.

[0036] In some of any embodiments, the integrated vector copy number (iVCN) of the CAR+T cells in the composition, on average, is between or between about 0.4 copies per diploid genome and 2.0 copies per diploid genome, inclusive.

[0037] In some of any embodiments, the integrated vector copy number (iVCN) of the CAR+T cells in the composition, on average, is between or between about 0.8 copies per diploid genome and 2.0 copies per diploid genome, inclusive. In some of any embodiments, the integrated vector copy number (iVCN) of the CAR+T cells in the composition, on average, is between or between about 0.8 copies per diploid genome and 1.0 copies per diploid genome, inclusive. In some of any embodiments, the integrated vector copy number (iVCN) of the CAR+T cells in the composition, on average, is between or between about 1.0 copies per diploid genome and 1.5 copies per diploid genome, inclusive. In some of any embodiments, the integrated vector copy number (iVCN) of the CAR+T cells in the composition, on average, is between or between about 1.5 copies per diploid genome and 2.0 copies per diploid genome, inclusive.

[0038] In some of any embodiments, at or prior to the administration of the composition of engineered T cells, the subject has received at least 3 prior antimyeloma treatment regimens. In some of any embodiments, at or prior to the administration of the composition of engineered T cells, the subject has received three or more therapies, optionally four or more prior therapies, selected from among an autologous stem cell transplant (ASCT); an immunomodulatory agent; a proteasome inhibitor; and an anti-CD38 agent, unless the subject was not a candidate for or was contraindicated for one or more of the therapies. In some of any embodiments, at or prior to the administration of the composition of engineered T cells, the subject has received three or more therapies, optionally four or more prior therapies, optionally selected from among an autologous stem cell transplant (ASCT); an immunomodulatory agent and a proteasome inhibitor, either alone or in combination; and an anti-CD38 agent. In some of any embodiments, at or prior to the administration of the composition of engineered T cells, the subject has received all three of the following therapies including autologous stem cell transplant (ASCT); a regimen including an immunomodulatory agent and a proteasome inhibitor; and an anti-CD38 agent.

[0039] In some of any embodiments, at or prior to the administration of the composition comprising engineered T cells, the subject has received all three of the following antimyeloma treatment regimens: autologous stem cell transplant (ASCT); an immunomodulatory agent and / or a proteasome inhibitor, either alone or in combination; and an anti-CD38 agent. In some of any embodiments, induction with or without bone marrow transplant and with or without maintenance therapy is considered one regimen for purpose of determining the number of prior antimyeloma treatment regimens.

[0040] In some of any embodiments, at or prior to the administration of the composition comprising engineered T cells, the subject is refractory to the last antimyeloma treatment regimen. In some of any embodiments, refractory myeloma is defined as documented progressive disease during or within 12months, measured from the last dose, of completing treatment with the last anti-myeloma treatment regimen. In some of any embodiments, refractory myeloma is defined as documented progressive disease during or within 60 days, measured from the last dose, of completing treatment with the last anti- myeloma treatment regimen.

[0041] In some of any embodiments, the immunomodulatory agent is selected from among thalidomide, lenalidomide, and pomalidomide, either alone or in combination. In some of any embodiments, the proteasome inhibitor is selected from among bortezomib, carfilzomib, and ixazomib, either alone or in combination.

[0042] In some of any embodiments, the subject has undergone at least one complete cycle of treatment with the antimyeloma treatment regimen comprising the immunomodulatory agent and / or the proteasome inhibitor unless progressive disease was the best response to the antimyeloma treatment regimen. In some of any embodiments, the subject has undergone at least two consecutive cycles of treatment with the antimyeloma treatment regimen comprising the immunomodulatory agent and / or the proteasome inhibitor unless progressive disease was the best response to the antimyeloma treatment regimen.

[0043] In some of any embodiments, the anti-CD38 agent is an anti-CD38 antibody. In some of any embodiments, the anti-CD38 agent is or comprises daratumumab. In some of any embodiments, the anti- CD38 agent is used as part of a combination regimen or as a monotherapy.

[0044] In some of any embodiments, at the time of the administration of the composition comprising engineered T cells, the subject has not had an active or a history of plasma cell leukemia (PCL). In some of any embodiments, at the time of the administration, the subject has relapsed or has been refractory following at least 3 or at least 4 prior antimyeloma treatment regimen. In some of any embodiments, at the time of the administration, the subject has a time from diagnosis of multiple myeloma of approximately 4 years or between 2 and 15 years or between 2 and 12 years. In some of any embodiments, at the time of the administration, the subject has received about 10 or between 3 and 15 or between 4 and 15 prior antimyeloma treatment regimen. In some of any embodiments, at the time of the administration, the subject has been refractory to or not responded to bortezomib, carfilzomib, lenalidomide, pomalidomide, and / or an anti-CD38 monoclonal antibody. In some of any embodiments, at the time of the administration, the subject has had a prior autologous stem cell transplant. In some of any embodiments, at the time of the administration, the subject has not had a prior autologous stem cell transplant (ASCT) due to ineligibility for ASCT, optionally ineligibility due to age or other documented reasons. In some of any embodiments, at the time of the administration, the subject has IMWG high risk cytogenetics. In some of any embodiments, the subject does not have a central nervous system involvement of MM, plasma cell leukemia, Waldenstrom’s macroglobulinemia, POEMS (polyneuropathy, organomegaly, endocrinopathy, monoclonal protein, skin changes) syndrome, and / orclinically significant amyloidosis. In some of any embodiments, the subject has not received a prior CAR T cell or genetically-modified T cell therapy. In some of any embodiments, the subject has not received a prior BCMA-targeted therapy such as an anti-BCMA monoclonal antibody or bispecific antibody.

[0045] In some of any embodiments, the method further comprises obtaining a leukapheresis sample from the subject for manufacturing the composition comprising engineered T cells.

[0046] In some of any embodiments, the subject has not received a therapeutic dose of a corticosteroid, optionally within at or about 14 days prior to the time of leukapheresis. In some of any embodiments, the subject has not received an immunosuppressive therapy within 4 weeks of leukapheresis, optionally wherein the immunosuppressive therapy comprises a a calcineurin inhibitor, methotrexate or other chemotherapeutics, mycophenolate, rapamycin, immunosuppressive antibodies such as anti-TNF, anti-IL6, or anti-IL6R. In some of any embodiments, the subject has not received a autologous stem-cell transplant within at or about 6 months prior to the time of leukapheresis.

[0047] In some of any embodiments, the subject has not achieved complete remission (CR) in response to a prior therapy. In some of any embodiments, the subject has not achieved an objective response (partial response (PR) or better) in response to a prior therapy. In some of any embodiments, the subject is or has been identified as having an Eastern Cooperative Oncology Group Performance Status (ECOG PS) of 0 or 1.

[0048] In some of any embodiments, the CAR may include an extracellular antigen-binding domain, including a variable heavy chain (VH) including a heavy chain complementarity determining region 1 (CDR-H1), a heavy chain complementarity determining region 2 (CDR-H2) and a heavy chain complementarity determining region 3 (CDR-H3) contained within the sequence set forth in SEQ ID NO: 116 and a variable light chain (VL) including a light chain complementarity determining region 1 (CDR- L1), a light chain complementarity determining region 2 (CDR-L2) and a light chain complementarity determining region 3 (CDR-L3) contained within the sequence set forth in SEQ ID NO: 119; a VHincluding a CDR-H1, a CDR-H2 and a CDR-H3 sequences set forth in SEQ ID NOS:97, 101 and 103, respectively, and a VLincluding a CDR-L1, a CDR-L2 and a CDR-L3 sequences set forth in SEQ ID NOS:105, 107 and 108, respectively; a VHincluding a CDR-H1, a CDR-H2 and a CDR-H3 sequences set forth in SEQ ID NOS:96, 100 and 103, respectively, and a VLincluding a CDR-L1, a CDR-L2 and a CDR-L3 sequences set forth in SEQ ID NOS:105, 107 and 108, respectively; a VHincluding a CDR-H1, a CDR-H2 and a CDR-H3 sequences set forth in SEQ ID NOS:95, 99 and 103, respectively, and a VLincluding a CDR-L1, a CDR-L2 and a CDR-L3 sequences set forth in SEQ ID NOS: 105, 107 and 108, respectively; a VHincluding a CDR-H1, a CDR-H2 and a CDR-H3 sequences set forth in SEQ ID NOS:94, 98 and 102, respectively, and a VL including a CDR-L1, a CDR-L2 and a CDR-L3 sequences set forth in SEQ ID NOS: 104, 106 and 108, respectively; or a VHincluding the amino acid sequence of SEQ ID NO: 116 and a VLincluding the amino acid sequence of SEQ ID NO: 119; and a spacerincluding an IgG4 / 2 chimeric hinge or a modified IgG4 hinge; an IgG2 / 4 chimeric CH2 region; and an IgG4 CH3 region, which optionally is about 228 amino acids in length; optionally wherein the spacer is set forth in SEQ ID NO: 174; and a transmembrane domain, optionally a transmembrane domain from a human CD28; and an intracellular signaling region including a cytoplasmic signaling domain of a CD3- zeta (CD3ζ) chain and a costimulatory signaling region including an intracellular signaling domain of a T cell costimulatory molecule or a signaling portion thereof. In some embodiments, the spacer is the spacer set forth in SEQ ID NO:174.

[0049] In some of any embodiments, the VH is or may include the amino acid sequence of SEQ ID NO: 116; and the VL is or includes the amino acid sequence of SEQ ID NO: 119. In some of any embodiments, the extracellular antigen-binding domain may include an scFv. In some of any embodiments, the VH and the VL are joined by a flexible linker. In some of any embodiments, the scFv may include a linker including the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:1). In some of any embodiments, the VH is carboxy-terminal to the VL. In some of any embodiments, the scFv comprises a linker comprising the amino acid sequence SRGGGGSGGGGSGGGGSLEMA (SEQ ID NO:255)

[0050] In some of any embodiments, the extracellular antigen-binding domain may include the amino acid sequence of SEQ ID NO: 114 or 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 SEQ ID NO: 114. In some of any embodiments, the extracellular antigen-binding domain may include the amino acid sequence of SEQ ID NO: 114.

[0051] In some of any embodiments, a nucleic acid encoding the extracellular antigen-binding domain may include the sequence of nucleotides of SEQ ID NO:113; a sequence of nucleotides that has at least 90% sequence identity thereto; or a degenerate sequence of either of the preceding sequences. In some of any embodiments, the nucleic acid encoding the extracellular antigen-binding domain may include the sequence of nucleotides of SEQ ID NO:115. In some of any embodiments, the VHis amino- terminal to the VL.

[0052] In some of any embodiments, the cytoplasmic signaling domain is or may include the sequence set forth in SEQ ID NO:143 or a sequence of amino acids that has at least 90% sequence identity to SEQ ID NO:143. In some of any embodiments, the costimulatory signaling region may include an intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof. In some of any embodiments, the costimulatory signaling region may include an intracellular signaling domain of 4-1BB, optionally human 4-1BB. In some of any embodiments,the costimulatory signaling region is or may include the sequence set forth in SEQ ID NO:4 or a sequence of amino acids that has at least 90% sequence identity to the sequence set forth in SEQ ID NO: 4. In some of any embodiments, thecostimulatory signaling region is between the transmembrane domain and the cytoplasmic signaling domain of a CD3-zeta (CD3ζ) chain.

[0053] In some of any embodiments, the transmembrane domain is or may include a transmembrane domain from human CD28. In some of any embodiments, the transmembrane domain is or may include the sequence set forth in SEQ ID NO:138 or a sequence of amino acids that has at least 90% sequence identity to SEQ ID NO:138.

[0054] In some of any embodiments, the CAR may include from its N to C terminus in order: the extracellular antigen-binding domain, the spacer, the transmembrane domain and the intracellular signaling region. In some of any embodiments, the CAR may include an extracellular antigen-binding domain, including a variable heavy chain (VH) including a heavy chain complementarity determining region 1 (CDR-H1), a heavy chain complementarity determining region 2 (CDR-H2) and a heavy chain complementarity determining region 3 (CDR-H3) contained within the sequence set forth in SEQ ID NO: 116 and a variable light chain (VL) including a light chain complementarity determining region 1 (CDR- L1), a light chain complementarity determining region 2 (CDR-L2) and a light chain complementarity determining region 3 (CDR-L3) contained within the sequence set forth in SEQ ID NO: 119; a spacer including a modified IgG4 hinge; an IgG2 / 4 chimeric CH2 region; and an IgG4 CH3 region, that is about 228 amino acids in length; a transmembrane domain from a human CD28; and an intracellular signaling region including a cytoplasmic signaling domain of a CD3-zeta (CD3ζ) chain and a costimulatory signaling region including an intracellular signaling domain of a 4-1BB.

[0055] In some of any embodiments, the CAR may include an extracellular antigen-binding domain, including the sequence set forth in SEQ ID NO: 114 or a sequence of amino acids having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 114; a spacer including the sequence set forth in SEQ ID NO: 174 or a sequence of amino acids that has at least 90% sequence identity to SEQ ID NO:174; a transmembrane domain including the sequence set forth in SEQ ID NO:138 or a sequence of amino acids that has at least 90% sequence identity to SEQ ID NO:138; and an intracellular signaling region including a cytoplasmic signaling including the sequence set forth in SEQ ID NO:143 or a sequence of amino acids that has at least 90% sequence identity to SEQ ID NO:143 and a costimulatory signaling region including the sequence set forth in SEQ ID NO:4 or a sequence of amino acids that has at least 90% sequence identity to the sequence set forth in SEQ ID NO: 4.

[0056] In some of any embodiments, the CAR may include an extracellular antigen-binding domain, including the sequence set forth in SEQ ID NO: 114; a spacer including the sequence set forth in SEQ ID NO: 174; a transmembrane domain including the sequence set forth in SEQ ID NO:138; and an intracellular signaling region including a cytoplasmic signaling including the sequence set forth in SEQ ID NO:143 and a costimulatory signaling region including the sequence set forth in SEQ ID NO:4. In some of any embodiments, the CAR may include an extracellular antigen-binding domain set forth inSEQ ID NO: 114; a spacer set forth in SEQ ID NO: 174; a transmembrane domain set forth in SEQ ID NO:138; and an intracellular signaling region including a cytoplasmic signaling set forth in SEQ ID NO:143 and a costimulatory signaling region set forth in SEQ ID NO:4. In some of any embodiments, CAR may include the sequence set forth in SEQ ID NO:19. In some of any embodiments, the sequence of the CAR is set forth in SEQ ID NO:19.

[0057] In some of any embodiments, following expression of a polynucleotide encoding the CAR in a human cell, optionally a human T cell, the transcribed RNA, optionally messenger RNA (mRNA), from the polynucleotide, exhibits at least 70%, 75%, 80%, 85%, 90%, or 95% RNA homogeneity. In some of any embodiments, the CAR is encoded by a polynucleotide sequence comprising the sequence set forth in SEQ ID NO: 13 or a sequence that exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In some of any embodiments, the CAR is encoded by a polynucleotide sequence comprising the sequence set forth in SEQ ID NO: 13.

[0058] In some of any embodiments, the binding of the extracellular antigen-binding domain and / or the CAR, or a measure indicative of function or activity of the CAR following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of BCMA. In some of any embodiments, the concentration or amount of the soluble or shed form of the BCMA corresponds to a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient, or on average in a multiple myeloma patient population, or at a concentration or amount of the soluble or shed BCMA at which the binding or measure is reduced or blocked, or is substantially reduced or blocked, for cells expressing a reference anti-BCMA recombinant receptor, optionally a reference anti-BCMA CAR, in the same assay.

[0059] In some of any embodiments, prior to the administration, the subject has received a lymphodepleting therapy comprising the administration of fludarabine at or about 20-40 mg / m2body surface area of the subject, optionally at or about 30 mg / m2, daily, for 2-4 days, and / or cyclophosphamide at or about 200-400 mg / m2body surface area of the subject, optionally at or about 300 mg / m2, daily, for 2-4 days. In some of any embodiments, prior to the administration, the subject has received a lymphodepleting therapy comprising the administration of fludarabine at or about 20-40 mg / m2body surface area of the subject, optionally at or about 30 mg / m2, daily, for 2-4 days. In some of any embodiments, prior to the administration, the subject has received a lymphodepleting therapy comprising the administration of cyclophosphamide at or about 200-400 mg / m2body surface area of the subject, optionally at or about 300 mg / m2, daily, for 2-4 days. In some of any embodiments, the subject has received a lymphodepleting therapy comprising the administration of fludarabine at or about 30 mg / m2body surface area of the subject, daily, and cyclophosphamide at or about 300 mg / m2body surface area of the subject, daily, each for 3 days.

[0060] In some of any embodiments, the method is capable of achieving a specified response or outcome, optionally at a designated timepoint following initiation of the administration, in at least one of or in at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of subjects in a cohort of subjects having the MM, wherein: the response is selected from the group consisting of objective response (OR), complete response (CR), stringent complete response (sCR), very good partial response (VGPR), partial response (PR) and minimal response (MR); the response or outcome is or comprises an OR; and / or the response or outcome is or comprises a CR.

[0061] In some of any embodiments, the cohort of subjects has at least the same number of prior therapies, prognosis or prognostic factor, sub-type, secondary involvement or other specified patient characteristic or characteristics, as the subject treated by the method. In some of any embodiments, the response or outcome is durable for greater than at or about 3, 6, 9 or 12 months. In some of any embodiments, the response or outcome determined at or about 3, 6, 9 or 12 months after the designated timepoint is equal to or improved compared to the response or outcome determined at the designated timepoint.

[0062] In some of any embodiments, the response or outcome is or comprises or further comprises the absence of neurotoxicity, the absence of cytokine release syndrome (CRS), and / or the absence of macrophage activation syndrome / hemophagocytic lymphohistiocytosis (MAS / HLH). In some of any embodiments, the method does not result in a specified toxicity outcome, optionally at a designated timepoint following initiation of the administration, in at least one of or in at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM.

[0063] In some of any embodiments, the specified toxicity outcome is neurotoxicity, cytokine release syndrome (CRS), and / or macrophage activation syndrome / hemophagocytic lymphohistiocytosis (MAS / HLH). In some of any embodiments, the specified toxicity outcome is neurotoxicity, and neurotoxicity does not result in at least 60%, 70% or 80% of the subjects in the cohort of subjects having the MM. In some of any embodiments, the specified toxicity outcome is grade 3 or higher, or grade 4 or higher, neurotoxicity. In some of any embodiments, the specified toxicity outcome is grade 3 or higher neurotoxicity, and grade 3 or higher neurotoxicity does not result in at least 80%, 85%, 90% or 95% of the subjects in the cohort of subjects having the MM. In some of any embodiments, the specified toxicity outcome is cytokine release syndrome (CRS), optionally grade 3 or higher, or grade 4 or higher, cytokine release syndrome (CRS). In some of any embodiments, the CRS does not result in at least 15%, 20%, 25% or 30% of the subjects in the cohort of subjects having the MM. In some of any embodiments, the designatied timepoint is at or about or within 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 12 days, 13 days, 14 days or 15 days following initiation of administration. In some of anyembodiments, the designated timepoint is at or about 1 month, 3 months, 6 months, 9 months, or 12 months following initiation of the administration.

[0064] In some of any embodimetns, the method does not result in any cytokine release syndrome (CRS) in at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM . In some of any embodiemtns, the method does not result in severe cytokine release syndrome (CRS) in at least at least at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM . In some of any embodiments, the method does not result in any neurotoxicity in at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM. In some of any embodimetns, the method does not result in severe neurotoxicity in at least at least at least 70%, at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM . In some of any embodiments, the method does not result in severe CRS and severe neurotoxicity in at least at least at least 70%, at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM. In some of any embodiments, the method does not result in severe CRS and severe neurotoxicity in at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM. In any of such embodiment, the severe CRS is grade 3 or higher, grade 4 or higher or grade 5 CRS. In any of such embodiments, the severe neurotoxicity is grade 3 or higher, grade 4 or higher or grade 5 CRS.

[0065] In some of any embodiemtns, the administration of the composition is carried out on an outpatient basis, optionally unless or until the subject exhibits a sustained fever or a fever that is or has not been reduced or not reduced by more than 1ºC after treatment with an antipyretic. In some of any embodimetns, the administration of the composition is without admitting the subject to a hospital and / or without an overnight stay at a hospital, optionally unless or until the subject exhibits a sustained fever or a fever that is or has not been reduced or not reduced by more than 1ºC after treatment with an antipyretic. In some of any embodiments, the administration of the composition is without requiring admission to or an overnight stay at a hospital, optionally unless or until the subject exhibits a sustained fever or a fever that is or has not been reduced or not reduced by more than 1ºC after treatment with an antipyretic.

[0066] In some of any embodiments, the composition comprising engineered T cells is administered parenterally, optionally intravenously. In some of any embodiments, the subject is a human subject.

[0067] In some of any embodiments, the composition comprising engineered T cells is produced by a manufacturing process comprising: (i) exposing an input composition comprising primary T cells with a stimulatory reagent comprising an oligomeric particle reagent comprising a plurality of streptavidin mutein molecules under conditions to stimulate T cells, thereby generating a stimulated population, wherein: the oligomeric particle reagent comprises a first agent comprising an anti-CD3 antibody or antigen binding fragment thereof and a second agent comprising an anti-CD28 antibody or antigenbinding fragment thereof; (ii) introducing into T cells of the stimulated population, a heterologous polynucleotide encoding the CAR that targets BCMA, thereby generating a population of transformed cells; (iii) incubating the population of transformed cells for up to 96 hours; and (iv) harvesting T cells of the population of transformed cells, thereby producing a composition of engineered cells, wherein the harvesting is carried out at a time between 24 and 120 hours, inclusive, after the exposing to the stimulatory reagent is initiated. In some embodiments, the input composition autologous T cells selected from the subject, such as enriched by immunoaffinity-based selection for CD3 T cells or CD4 and CD8 T cells from a blood or apheresis (e.g. leukarephesis) sample from the subject.

[0068] In some of any embodiments, the composition may include engineered T cells is produced by a manufacturing process including exposing an input composition including primary T cells with a stimulatory reagent including an oligomeric particle reagent including a plurality of avidin, streptavidin, avidin mutein, or streptavidin mutein molecules under conditions to stimulate T cells, thereby generating a stimulated population, wherein the stimulatory reagent is capable of activating one or more intracellular signaling domains of one or more components of a TCR complex and one or more intracellular signaling domains of one or more costimulatory molecules. In some of any embodiments, the manufacturing process may further include introducing into T cells of the stimulated population, a heterologous polynucleotide encoding the CAR that targets BCMA, thereby generating a population of transformed cells. In some of any embodiments, the manufacturing process may further include incubating the population of transformed cells for up to 96 hours. In some of any embodiments, the incubating is carried out in basal media lacking one or more recombinant cytokines.

[0069] In some of any embodiments, the oligomeric particle reagent comprises a first agent comprising an anti-CD3 antibody or antigen binding fragment thereof and a second agent comprising an anti-CD28 antibody or antigen binding fragment thereof. In some of any embodiments, the anti-CD3 antibody or antigen binding fragment is a Fab and the anti-CD28 antibody or antigen binding fragment is a Fab. In some of any embodiments, the first agent and the second agent each comprise a streptavidin- binding peptide that reversibly binds the first agent and the second agent to the oligomeric particle reagent, optionally wherein the streptavidin-binding peptide comprises the sequence of amino acids set forth in any of SEQ ID NOS:266-270. In some of any embodiments, the streptavidin mutein molecule is a tetramer of a streptavidin mutein comprising amino acid residues Val44-Thr45-Ala46-Arg47 or Ile44- Gly45-Ala46-Arg47, optionally wherein the streptavidin mutein comprises the sequence set forth in any of SEQ ID NOS: 257, 272, 275, 277, 279, 273 or 276. In some of any embodiments, the oligomeric particle reagent comprises between 1,000 and 5,000 streptavidin mutein tetramers, inclusive. In some of any embodiments, the method further comprises, prior to harvesting the cells, adding biotin or a biotin analog after or during the incubation.

[0070] In some of any embodiments, the manufacturing process further includes harvesting T cells of the transformed population, thereby producing a composition of engineered cells. In some of any embodiments, the harvesting is carried out at a time between 24 and 120 hours, inclusive, after the exposing to the stimulatory reagent is initiated. In some of any embodiments, the harvesting is carried out at a time between 48 and 120 hours, inclusive, after the exposing to the stimulatory reagent is initiated. In some of any embodiments, the harvesting is carried out at a time when integrated vector is detected in the genome but prior to achieving a stable integrated vector copy number (iVCN) per diploid genome. In some of any embodiments, the harvesting is carried out at a time before the total number of viable cells at the harvesting is more than or more than about three times as the number of total viable cells of the stimulated population. In some of any embodiments, the harvesting is carried out at a time when the total number of viable cells at the harvesting is at or about three times, at or about two times, or the same or about the same as the number of total viable cells of the stimulated population. In some of any embodiments, the harvesting is carried out at a time when the percentage of CD27+CCR7+cells is greater than or greater than about 50% among total T cells in the population, total CD3+T cells in the population, total CD4+T cells in the population, or total CD8+T cells, or of CAR-expressing cells thereof, in the population. In some of any embodiments, the harvesting is carried out at a time when the percentage of CD45RA+CCR7+and CD45RA-CCR7+cells is greater than or greater than about 60% among total T cells in the population, total CD3+T cells in the population, total CD4+T cells in the population, or total CD8+T cells, or of CAR-expressing cells thereof, in the population.

[0071] In some of any embodiments, the cells in the administered composition are produced by a manufacturing process to produce an output composition exhibiting a predetermined feature, wherein iterations of the manufacturing process produce a plurality of the output compositions, optionally from human biological samples, when carried out among a plurality of different individual subjects, in which the predetermined feature of the output composition among the plurality of output compositions is selected from the mean percentage of cells of a memory phenotype in the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%; the mean percentage of cells of a central memory phenotype in the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%; the mean percentage of cells that are CD27+, CD28+, CCR7+, CD45RA-, CD45RO+, CD62L+, CD3+, CD95+, granzyme B-, and / or CD127+ in the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%; the mean percentage of cells that are CCR7+ / CD45RA- orCCR7+ / CD45RO+ in the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%; the mean percentage of central memory CD4+ T cells in the engineered CD4+ T cells, optionally CAR+CD4+ T cells, of the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%; the mean percentage of central memory CD8+ T cells in the engineered CD8+ T cells, optionally CAR+CD8+ T cells, of the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%; and / or the mean percentage of central memory T cells, optionally CD4+ central memory T cells and CD8+ central memory T cells, in the engineered T cells, optionally CAR+ T cells, of the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%.

[0072] In some of any embodiments, the administered composition is produced by a manufacturing process to produce an output composition exhibiting a predetermined feature, optionally a threshold number of cells expressing the CAR in the output composition, in at least about 80%, about 90%, about 95%, about 97%, about 99%, about 100%, or is 100% of the human biological samples in which it is carried out among a plurality of different individual subjects. In some of any embodiments, the composition including genetically engineered cells does not contain residual beads from the manufacturing process.

[0073] In some of any embodiments, the MM is a relapsed and / or refractory multiple myeloma (r / r MM).

[0074] Also provided herein is an article of manufacture including a composition including genetically engineered cells expressing a chimeric antigen receptor (CAR) that targets BCMA, and instructions for administering the composition of the cells in accordance with the method of any of the methods provided herein. Brief Description of the Drawings

[0075] FIG.1 shows exemplary quantifications determined by flow cytometry of cell purity of T cell compositions produced from non-expanded engineering processes using different donor types (Reference, Patient). Cells were engineered to express an anti-BCMA CAR (BCMA) or were mock transduced (mock). Percentages of CD3+ cells of live CD45+ cells (left panel), percentages of NK cellsof live CD45+ cells (middle panel), and percentages of CD19+ cells of live CD45+ cells (right panel) were determined.

[0076] FIG.2- FIG.3 shows exemplary quantifications of cell phenotypes determined by flow cytometry for expanded and non-expanded engineering processes using different donor types (Reference, Patient). Cells were engineered to express an anti-BCMA CAR (BCMA) or were mock transduced (mock). FIG.2 shows percentages of CD3+CD8+ and CD3+CD4+ cells of live CD45+ cells (left panel) and percentages of CD8+CAR+ and CD4+CAR+ cells of live CD45+ cells (right panel). FIG.3 shows ratios of CD4+ cells to CD8+ cells, and CD4+CAR+ cells to CD8+CAR+ cells.

[0077] FIG.4 shows exemplary quantifications determined by flow cytometry of cell viability of T cell compositions produced from non-expanded engineering processes using different donor types (Reference, Patient). Cells were engineered to express an anti-BCMA CAR (BCMA) or were mock transduced (mock). Percentages of aCas3+ cells of CD3+ cells were determined.

[0078] FIG.5A shows exemplary relationship between copy number per cell among total cells as assessed by standard VCN (without PFGE) and iVCN (with PFGE), in cell compositions produced from primary T cells from different human donors that had been engineered to express a CAR using an expanded process (○) or a non-expanded process (●). FIGS.5B-5C show the relationship between the copy number per cell in the cell compositions as assessed by standard VCN (FIG.5B) or iVCN (FIG. 5C) and the surface expression of the CAR, as indicated by the percentage of CAR-expressing CD3+ cells (%CD3+CAR+) among viable CD45+ cells assessed by flow cytometry.

[0079] FIGS.6A-6B show exemplary percentages of cell phenotypes resulting from expanded and non-expanded engineering processes using different donor types (Reference, Patient). Cells were engineered to express an anti-BCMA CAR (BCMA) or were mock transduced (mock). FIG.6A shows exemplary percentages of CD45RA+CCR7+ cells of aCas-CD8+CAR+ and aCas-CD4+CAR+ cells (left top panel), CD45RA-CCR7+ cells of aCas-CD8+CAR+ and aCas-CD4+CAR+ cells (right top panel), CD45RA-CCR7- cells of aCas-CD8+CAR+ and aCas-CD4+CAR+ cells (left bottom panel), and CD45RA+CCR7- cells of aCas-CD8+CAR+ and aCas-CD4+CAR+ cells (right bottom panel). FIG.6B shows exemplary percentages of CD27+CCR7+ cells of aCas-CD8+CAR+ and aCas-CD4+CAR+ cells.

[0080] FIG.7 shows exemplary proportions of T-cell memory phenotypes defined by surface expression of CD45RA and CCR7 on CAR T cells derived from donor-matched non-expanded process products and expanded process products. CAR T cells were produced from CD4+ and CD8+ T cells from one healthy donor (HD1) or 3 patients with multiple myeloma (MM1, MM2, or MM3).

[0081] FIGS.8A-8C show exemplary in vitro proliferative capacity of cells generated from non- expanded and expended processes following long-term anti-BCMA CAR-dependent stimulation with an agonistic antibody. CAR T cells were produced from CD4+ and CD8+ T cells from one healthy donor (HD1) or from 3 patients with multiple myeloma (MM1, MM2, MM3), and the numbers of total livecells were determined every 2 days following stimulation with microbeads coated with an agonistic antibody for 10 days. FIG.8A shows fold change in expansion calculated by dividing the daily counts by the starting number of cells. FIG.8B shows CAR T-cell counts transformed into an AUC for comparison between products (non-expanded process products; expanded process products; mock) or donors. Statistical significance was determined with a Mann-Whitney test; * p < 0.05. FIG.8C shows fold expansion of each donor calculated by dividing the daily fold expansion in the non-expanded process product group by the donor-matched expanded process product values.

[0082] FIGS.9A-9E show exemplary quantifications of intracellular IL-2, IFNγ, or TNF cytokine production measured by flow cytometry (FIGS.9A-9D) and secreted cytokines (FIG.9E) from anti- BCMA CAR T cells derived from donor-matched non-expanded process products and expanded process products. CAR T cells derived from one patient with multiple myeloma or from 2 healthy donors after culturing cells with an agonistic antibody for 5 hours. Frequency of CAR-positive cells expressing single cytokines (FIGS.9A-9C) or Boolean logic gated triple-positive cells (FIG.9D) within the CD4+CAR+ or CD8+CAR+ populations are shown. Cytokine protein secretion was measured by multiplex immunoassay quantitation of secreted cytokine concentrations (FIG.9E, showing the total protein secretion measured in culture supernatants) after culturing cells for 24 hours with MM.1S BCMA- positive target cells. Statistical significance was assessed by Mann-Whitney; * p < 0.05.

[0083] FIG.10A shows exemplary cytolytic potential of anti-BCMA CAR T cells engineered by non-expanded or expanded processes at different effector to target ratios. Area under the curve (AUC) values were compared either for individual arms (left panel of FIG.10B) or by manufacturing process (right panels of FIG.10B, statistical significance with a Mann-Whitney test; *p < 0.05).

[0084] FIGS.11A-11B show exemplary tumor burden and circulating CAR-T cells in the OPM-2 myeloma model over time following treatment with anti-BCMA CAR-T cell compositions generated from non-expanded and expanded matched-donor engineering processes. Tumor growth from Day -1 (before treatment) to about Day 53 post-treatment is shown in BLI (photons / second; y-axis) (FIG.11A) or calculated from area under the curve (AUC) of BLI for each group (FIG. 11B). Differences were compared using Mann-Whitney U test; *p < 0.05.

[0085] FIGS.12A-12B show exemplary CAR T kinetics and circulating CAR-T cell numbers in the OPM-2 myeloma model over time following treatment with anti-BCMA CAR-T cell compositions generated from non-expanded and expanded matched-donor engineering processes. FIG.12A shows circulating anti-BCMA CAR-T cell counts per 1 µl of blood post-treatment for each group. FIG.12B shows circulating anti-BCMA CAR-T cell counts per 1 µl of blood at the indicated time points post- treatment for each group (non-expanded, NE; expanded, E). Differences were compared using Mann- Whitney U test; *p < 0.05.Detailed Description

[0086] Provided herein are methods of using and uses of engineered cells expressing anti-BCMA recombinant receptors (e.g. CARs) and pharmaceutical compositions and formulations thereof, such as in the treatment of diseases, conditions, and disorders in which BCMA is expressed, most particularly a hematological malignancy that is a multiple myeloma (MM). In embodiments of the provided methods, the therapeutic T cell compositions containing the engineered cells are administered to a subject having MM, e.g., via adoptive cell therapy, such as adoptive T cell therapy. In particular embodiments of any of the provided methods, the T cells are engineered with a CAR that is directed against BCMA. In some aspects, the methods and uses provide for or achieve improved response and / or more durable responses or efficacy and / or a reduced risk of toxicity or other side effects, e.g., in particular groups of subjects treated, as compared to certain alternative methods. In some embodiments, the methods are advantageous by virtue of the administration of specified numbers or relative numbers of the engineered cells, the administration of cell compositions with predetermined features (e.g., ratios of particular types of the cells), the administration of cells of a particular high percentage of less differentiated cells (e.g. naïve- like or central memory cells or cells of an early differentiation state, such as CCR7+CD27+ cells), treatment of particular patient populations, such as those having a particular risk profile, staging, and / or prior treatment history, and / or combinations thereof.

[0087] In some embodiments, the methods and uses include administering to the subject T cells expressing genetically engineered (recombinant) cell surface receptors in adoptive cell therapy, which generally are chimeric receptors such as chimeric antigen receptors (CARs), recognizing BCMA expressed by, associated with and / or specific to the MM and / or cell type from which it is derived. The cells are generally administered in a composition formulated for administration; the methods generally involve administering one or more doses of the cells to the subject, which dose(s) may include a particular number or relative number of cells or of the engineered cells. In some cases, the BCMA- directed CAR+ engineered cells in the composition include a defined ratio or compositions of two or more sub-types within the composition, such as CD4 vs. CD8 T cells. In particular embodiments, the compositions of cells for use or administration in the provided methods include primary T cells engineered to express a BCMA-directed CAR that (i) contain a low percentage (e.g. less than 40%, less than 30%, less than 20%, or less than 10%) of exhausted cells and / or cells that display markers or phenotypes associated with exhaustion; and / or (ii) contain a relatively high percentage (e.g. greater than 50%, greater than 60%, greater than 70%, greater than 80% or greater than 90%) of memory-like T cells, such as naïve-like T cells, central memory T cells or long-lived memory T cells. In provided embodiments, the features of the compositions and provided methods result in improved or enhanced survival, expansion, persistence, and / or anti-tumor activity compared to methods involving administration other BCMA-directed CAR T cell therapies that contain a higher percentage of exhaustedcells and / or a higher number of cells that display phenotypes associated with exhaustion and / or that contain a lower percentage of certain T cells, such as naïve-like T cells, central memory T cells or long- lived memory T cells. In provided embodiments, the features of the compositions and provided methods result in improved therapeutic efficacy, e.g. increased percentage of patients achieving a complete response (CR), compared to methods involving administration of other BCMA-directed CAR T cell therapies that contain a higher percentage of exhausted cells and / or a higher number of cells that display phenotypes associated with exhaustion and / or that containa lower percentage of certain T cells, such as naïve-like T cells, central memory T cells or long-lived memory T cells. In provided methods, the features of the compositions and provided methods result in improved clinical durability of therapeutic response, such as CR, e.g., response that persists after a period of time from initiation of therapy, compared to methods involving administration of other BCMA-directed CAR T cell therapies that contain a higher percentage of exhausted cells and / or a higher number of cells that display phenotypes associated with exhaustion and / or that contain a lower percentage of memory-like T cells, such as naïve- like T cells, central memory T cells or long-lived memory T cells. In particular embodiments, the use or administration of the provided BCMA-directed CAR T cell compositions in the provided methods can be achieved with doses of cells that are more than 2-fold lower, such as 5-fold or 10-fold, lower than doses of reference BCMA-directed CAR T cell compositions (e.g. engineered with the same or similar CAR, such as with the same antigen-binding domain) but in which the reference BCMA-directed CAR T cell composition contain a higher percentage of exhausted cells and / or a higher number of cells that display phenotypes associated with exhaustion and / or that contains a lower percentage of memory-like T cells, such as naïve-like T cells, central memory T cells or long-lived memory T cells. In some embodiments, the reference BCMA-directed CAR T cell composition is a composition that is produced ex vivo by processes that involve steps of cultivating the cells under conditions for expansion, such as resulting in proliferation of cells or population doubling of cells (e.g.2, 3, 4, 5, 6, 7, 8, 9, 10 or more doublings of cells in the population compared to the start of the process) during the process for producing the cells.

[0088] In some embodiments, the BCMA-directed CAR T cell compositions for use in the provided methods and uses are produced by a relatively short process that do not include a step for cultivating the cells under conditions for expansion designed for expanding or proliferating the cells. Different processes are available for generating compositions containing genetically engineered T cell populations, including for generating engineered T cells that express a CAR, which typically include a step designed for or for the purpose of cultivating the cells to expand or increase proliferation of the cells. However, in particular aspects, some of these processes may require a long or a relatively long amount of time to generate the engineered cells. In addition, in various aspects, some existing processes may vary in the amount of time required to successfully produce engineered T cells suitable for cell therapy, making it difficult to coordinate that administration of the cell therapy. In certain aspects, some of theseprocesses may produce populations of cells that include a relatively high percentage or amount of exhausted cells, differentiated cells, or cells with a low potency. The provided BCMA-directed CAR T cell compositions for use in the provided methods address one or more of these problems.

[0089] In particular embodiments, the provided methods are used in connection with a process for efficiently producing or generating engineered cells that are suitable for use in a cell therapy. In some embodiments, provided compositions containing BCMA-directed CAR engineered T cells are produced by a process without the need for any additional steps for expanding the cells, e.g. without an expansion unit operation and / or without steps intended to cause expansion of cells. In aspects of processes for producing BCMA-directed CAR T cell composition, the processes include one or more steps for stimulating and genetically engineering (e.g., transforming, transducing or transfecting) T cells to produce a population of engineered T cells that may be collected or formulated for use as a composition for cell therapy. In particular embodiments, the processes include a step of transducing cells with a viral vector (e.g. lentiviral vector) that contains a nucleic acid encoding the BCMA-directed CAR. In some aspects, the provided processes result in the stable integration of the heterologous nucleic acid (expressed from the viral vector) into the genome of the cells. In some aspects, the provided processes generate engineered BCMA-directed CAR T cells with enhanced potency as compared to engineered T cell compositions produced from alternative processes, such as those that involve expanding the cells.

[0090] In particular aspects, the durations of the processes for producing the provided compositions can be measured from when cells, e.g., T cells of an input cell population or input composition, are first contacted or exposed to stimulating conditions (e.g., as described herein such as in Section II-C), referred to herein as the initiation of the stimulation or stimulating and also referred to herein as the exposing to the stimulatory reagent, e.g., as in when the exposing to the stimulatory reagent is initiated. In some embodiments, the duration of time required to harvest or collect an output population (also referred to herein as an output composition or as a composition of engineered cells, e.g., engineered T cells) containing engineered cells is measured from initiation of the stimulation. In particular embodiments, the duration of the process is, is about, or is less than 120 hours, 108 hours, 96 hours, 84 hours, 72 hours, 60 hours, 48 hours, 36 hours, or 30 hours. In particular embodiments, the duration of the process is, is about, or is less than 5 days, 4 days, 3 days, 2 days, or one day. In particular embodiments, the engineered cells, e.g., the cells of the output composition or population, are more potent, persistent or naïve-like than cells that are engineered with processes that require longer amounts of time. In some aspects, the duration, e.g., the amount of time required to generate or produce an engineered population of T cells, of the provided processes are shorter than those of some existing processes by, by about, or by at least 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, or more than 7 days. In some embodiments, the duration of the provided process is, is about, or is less than 75%, 60%, 50%, 40%, 30%, 25%, 15%, or 10% of alternative or existing processes.

[0091] In certain embodiments, the provided processes are performed on a population of cells, e.g., CD3+, CD4+, and / or CD8+ T cells, that are isolated, enriched, or selected from a biological sample. In some aspects, the provided methods can produce or generate a composition of engineered T cells from when a biological sample is collected from a subject within a shortened amount of time as compared to other methods or processes. In some embodiments, the provided methods can produce or generate engineered T cells, including any or all times where biological samples, or enriched, isolated, or selected cells are cryopreserved and stored, within or within about 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, or 2 days, or within or within about 120 hours, 96 hours, 72 hours, or 48 hours, from when a biological sample is collected from a subject to when the engineered T cells are collected, harvested, or formulated (e.g., for cryopreservation or administration).

[0092] In particular embodiments, the processes for producing or engineering T cell populations include a step of stimulating the cells, such as prior to transduction with a viral vector. In aspects of the provided processes, stimulation is carried out with an oligomeric stimulatory reagent, such as a streptavidin mutein oligomer, to which is immobilized or attached a stimulatory binding agent(s), e.g. anti-CD3 / anti-CD28. Existing reagents for use in stimulating T cells in vitro, such as in the absence of exogenous growth factors or low amounts of exogenous growth factors, are known (see e.g. US Patent 6,352,694 B1 and European Patent EP 0700430 B1). In general, such reagents may employ beads, e.g., magnetic beads, of greater than 1 µm in diameter to which various binding agents (e.g. anti-CD3 antibody and / or anti-CD28 antibody) are immobilized. However, in some cases, such magnetic beads are, for example, difficult to integrate into methods for stimulating cells under conditions required for clinical trials or therapeutic purposes since it has to be made sure that these magnetic beads are completely removed before administering the expanded T cells to a subject. In some aspects, such removal, such as by exposing the cells to a magnetic field, may decrease the yield of viable cells available for the cell therapy. In certain cases, such reagents, e.g., stimulatory reagents containing magnetic beads, must be incubated with the cells for a minimal amount of time to allow a sufficient amount of detachment of the T cells from the stimulatory reagent.

[0093] The provided processes utilizing oligomeric stimulatory reagents, e.g. streptavidin mutein polymer, overcome such potential limitations. For example, in some embodiments, the provided processes avoid or reduce risk of residual stimulatory reagent, e.g., reagents containing magnetic beads, in the output cells generated or produced by the processes. In some embodiments, this also means that a process that is compliant with GMP standards can be more easily established compared to other methods, such as those where additional measures have to be taken to ensure that the final engineered T cell population is free of beads. In some embodiments, this may be readily accomplished in the present embodiments by the addition of a substance, e.g., a competition reagent, that dissociates the oligomeric stimulatory reagents from the cells, e.g., by simply rinsing or washing the cells. e.g., by centrifugation.Thus, in some aspects, removal or separation of oligomeric stimulatory reagent from cells, such as by the addition of a substance or competition reagent, results in little or no cell loss as compared to removal or separation of bead based stimulatory reagents. In some aspects, the timing of the oligomeric stimulatory reagent removal or separation is not limited or is less limited than the removal or separation of bead based stimulatory reagents. Thus, in some aspects, the oligomeric stimulatory reagent may be removed or separated from the cells at any time or stage during the provided processes.

[0094] In some aspects, the use of oligomeric stimulatory reagents (e.g. anti-CD3 / anti-CD28 streptavidin mutein oligomers) can result in an overall reduced stimulatory signal compared to alternative stimulatory reagents, such as anti-CD3 / anti-CD28 paramagnetic beads. The provided process, which can involve a weaker or reduced stimulation, can generate engineered CAR+ T cells that are as, or even more, potent, persistent, or efficacious as CAR+ T cells generated by processes that involve stronger stimulatory conditions or higher amounts or concentrations of stimulatory reagent, such as may occur following stimulation with anti-CD3 / anti-CD28 paramagnetic beads. In addition, in some embodiments, stimulating cells with a lower amount or relatively low amount of oligomeric stimulatory reagents may increase the potency, efficacy, or persistency of the resulting engineered cell population, as compared to processes using higher amounts of oligomeric stimulatory reagent. Such embodiments contemplate that such effects may persist even at doses sufficiently low enough to reduce the expression of activation markers or the portion of cells positive for the activation markers during and after the process.

[0095] In certain embodiments, the engineered T cells, e.g., output composition or populations of T cells containing T cells expressing a recombinant receptor, such as a chimeric antigen receptor, produced or generated by the provided processes are particularly effective or potent when utilized as cells for a cell therapy. For example, in some aspects, an output composition containing engineered T cells, e.g., CAR+ T cells, that are generated from the provided processes have a much higher degree of potency and / or proliferative capacity than engineered T cells generated or produced by alternative existing processes. In some aspects, an output composition containing engineered T cells, e.g., CAR+ T cells, produced by the provided processes have enhanced anti-tumor or anti-cancer cell activity than engineered T cells, e.g., CAR+ T cells, produced by alternative or existing methods.

[0096] In particular embodiments, the processes for producing the provided BCMA-directed T cell compositions that do not contain steps where the cells are expanded to a threshold amount or concentration have further advantages. In some aspects, protocols that do not rely on expanding the cells to increase the number or concentration of cells from a starting cell population, e.g., an input population, do not require incubations or cultivations that may vary between cell populations. For example, some embodiments contemplate that cell populations obtained from different subjects, such as subjects having different diseases or disease subtypes, particularly as is the case for patients with MM, including high- risk, aggressive and / or R / R MM, may divide or expand at different rates. In certain aspects, eliminatingpotentially variable steps requiring cell expansion allows for the duration of the whole process to be tightly controlled. In certain embodiments, the variability of the process duration is reduced or eliminated which may, in some aspects, allow for improved coordination for appointments and treatment between doctors, patients, and technicians to facilitate autologous cell therapies.

[0097] In some embodiments, the provided methods involve treating a specific group or subset of subjects, e.g., subjects identified as having high-risk disease, e.g., high-risk hematological malignancy or a high-risk MM. In some aspects, the methods treat subjects having a form of poor prognosis MM, such as MM that has relapsed or is refractory (R / R) to standard therapy and / or has a poor prognosis. In some aspects, the methods treat subjects having a MM that has relapsed or is refractory (R / R) to standard therapy. In particular aspects, the engineered cells are autologous to the subject and are administered following generation by ex vivo processes that are shortened compared to existing methods, that do not include or involve a cultivation step for expanding the cells during the methods of producing the engineered cells, and / or that are able to produce a CAR-engineered T cell composition that is less differentiated permitting administration of lower doses. As a result, the provided methods are advantageous compared to existing methods because they can shorten the time until the engineered T cell therapy is available to the patient, particularly among patients who are in need of treatment, such as subjects that have relapsed to or are refractory to treatment following one or more other prior therapies for treating the disease or condition. In some aspects, the provided methods, compositions, uses and articles of manufacture achieve improved and superior responses to available therapies. In some embodiments, the improved or superior responses are to current standard of care (SOC).

[0098] Multiple myeloma (MM) is a hematologic malignancy characterized by the clonal proliferation and accumulation of malignant plasma cells in the bone marrow and development of osteolytic lesions (Palumbo et al., N Engl J Med.2011; 364(11):1046-60). It accounts for approximately 10% of all hematologic malignancies. It is estimated that there will be approximately 32,110 new cases diagnosed and 12,960 deaths from MM in the United States (US) in 2019 (Siegel et al., CA Cancer J Clin.2019; 69(1):7-34).

[0099] The median age at diagnosis is 69 years, and less than 15% of those newly diagnosed are under the age of 55 years (SEER Cancer Stat Facts: Myeloma Web site, https: / / seer.cancer.gov / statfacts / html / mulmy.html., accessed March 8, 2019). Clinical features of symptomatic disease are summarized by the so-called “CRAB criteria,” which consist of calcium elevation, renal impairment, anemia, and lytic bone lesions or osteoporosis (Palumbo et al., N Engl J Med.2011; 364(11):1046-60). Multiple myeloma is a molecularly, biologically and clinically heterogeneous disease with some patients progressing rapidly despite treatment and others not requiring therapy for several years. Overall survival (OS) (median) is 5 to 6 years (Nandakumar et al., JCO 2019; 37:15_suppl: 8039).

[0100] Novel agents (i.e., immunomodulatory drugs, proteasome inhibitors, anti-CD38 or SLAMF7- directed monoclonal antibodies) alone or in combination with conventional therapies have led to significant improvements in the clinical outcomes of MM patients. However, despite these recent advancements, MM remains an incurable disease with multiple relapses and high mortality rates as drug resistant clones emerge (Cho et al., Front Immunol.2018; 9:1821; Cornell et al., Bone Marrow Transplant 2016; 51(4):479-91). The median overall survival (OS) of patients with MM who are relapsed and / or refractory (R / R) to available therapies is poor. Therefore, newer therapeutic approaches are needed to overcome relapse and improve the survival outcomes in patients with MM.

[0101] B-cell maturation antigen (BCMA), a member of the tumor necrosis factor (TNF) receptor superfamily, is a cell surface protein expressed on plasma cells that is involved in regulating the maturation of B cells and differentiation into plasma cells. It is induced during differentiation of plasma cells in parallel with the loss of expression of a related receptor for B-cell activation factor (BAFF-R). Binding of BCMA to its ligands, B- cell activation factor (BAFF) and a proliferation-inducing ligand (APRIL), leads to survival of plasma cells, resulting in enhanced humoral immunity.

[0102] BCMA is an attractive therapeutic target because BCMA is highly expressed on MM cell lines and on cells from patients with MM, and its expression appears to increase with progression of the disease (Tai et al., Immunotherapy 2015; 7(11):1187-99). Importantly, BCMA protein is not expressed in hematopoietic stem cells, naïve B cells, or normal non-hematopoietic tissues (Carpenter et al., Clin Cancer Res.2013; 19(8):2048-60; Tai et al., Immunotherapy 2015; 7(11):1187-99). Thus, toxicity associated with on-target / off-tumor interactions are reduced with agents targeting BCMA.

[0103] A challenge in CAR T cell development is to generate a product that consistently expands, persists, and mediates durable antitumor responses after infusion. BCMA-targeted CAR T cells are being evaluated for treatment of MM. In preclinical studies, T cells transduced with BCMA-targeted chimeric antigen receptor (CAR) construct produced high level of cytokines (e.g., interferon gamma [IFN-γ], TNFα, interleukin-2 [IL-2] and proliferated upon stimulation with BCMA-expressing target cells. In addition, BCMA-targeted CAR T cells killed BCMA-expressing MM cells and eradicated BCMA- expressing tumors in mouse xenograft models (Carpenter et al., Clin Cancer Res.2013; 19(8):2048-60). The persistence of CAR-engineered T cells and durability of response of patients with multiple myeloma following administration of BCMA-directed CAR T cells is a challenge.

[0104] In particular embodiments, the methods provided herein are based on administration of a BCMA-directed CAR T cell therapy in which the CAR contains a BCMA-directed scFv antigen binding domain. The CAR further contains an intracellular signaling domain containing a signaling domain from CD3zeta, and also incorporates a 4-1BB costimulatory domain.

[0105] The provided methods are based on findings that a lower differentiation state of adoptively transferred T cells can influence the ability of these cells to persist and promote durable antitumorimmunity. In some embodiments, the provided BCMA-directed CAR+ engineered T cell compositions are produced by a method in which the cells are not cultivated under conditions of expansion, thereby limiting or reducing the number of population doublings of the final engineered output composition and resulting in a less differentiated product. Yet, the provided compositions also are produced via processes that result in stably integrated vector copy number (iVCN) to ensure consistent and reliable expression of the CAR, thereby resulting in a consistent cell product for administration to subjects and low variability among CAR-expressing cells in administered doses. In contrast, most protocols for T cell engineering routinely expand T cells ex vivo for 9 to 14 days or more. Provided data exemplified herein support a model in which CAR T cell products with an increased composition of less differentiated memory T cells may exhibit enhanced durable antitumor activity. These findings reveal that strategies aimed at minimizing effector differentiation in CAR T cell products could result in improved clinical efficacy. Provided herein are embodiments that can meet such aims.

[0106] The observations herein support treating subjects with high-risk disease with a BCMA- directed CAR T cell therapy in accord with the provided methods. For example, subjects with MM, including patients with high-risk MM, such as those with relapsed / refractory (R / R) MM, can be treated in accord with the provided methods. In some embodiments, the provided methods can be used to treat subjects that have been heavily pretreated (e.g. with one, two, three, four, or more prior therapies for treating the disease).

[0107] All publications, including patent documents, scientific articles and databases, referred to in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication were individually incorporated by reference. If a definition set forth herein is contrary to or otherwise inconsistent with a definition set forth in the patents, applications, published applications and other publications that are herein incorporated by reference, the definition set forth herein prevails over the definition that is incorporated herein by reference.

[0108] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. I. METHODS AND USES OF BCMA-TARGETED CELL THERAPY IN MULTIPLE MYELOMA

[0109] Provided herein are methods of treatment that involve administering engineered cells or compositions containing engineered cells, such as engineered T cells. Also provided are methods and uses of provided BCMA-directed CAR engineered cells (e.g., T cells) and / or compositions thereof, including methods for the treatment of subjects having a multiple myeloma (MM), including high-risk MM, such as R / R MM, that involves administration of the engineered cells and / or compositions thereof. In some embodiments, the methods and use of provided BCMA-directed CAR engineered cells (e.g., Tcells) and / or compositions thereof, including methods for the treatment of subjects with R / R MM that have failed at least two or more prior therapies. In particular embodiments, the method includes administering to the subject a dose of T cells that includes CD4+ and CD8+ T cells, wherein the T cells comprises a chimeric antigen receptor (CAR) that specifically binds to BCMA.

[0110] In some embodiments, the methods and uses include administering to the subject cells expressing genetically engineered (recombinant) cell surface receptors in adoptive cell therapy, which generally are chimeric receptors such as chimeric antigen receptors (CARs), recognizing BCMA expressed by, associated with and / or specific to the MM and / or cell type from which it is derived. The cells are generally administered in a composition formulated for administration. In some embodiments, cells are collected from the subject prior to treatment for the purpose of engineering the cells with the BCMA-directed recombinant receptor (e.g. CAR). In some embodiments, the cells are collected by leukapheresis. In some aspects, the cells are engineered by ex vivo methods that do not involve cultivating the cells for expansion (hereinafter also called non-expanded process). Exemplary non-expanded processes for engineering the provided CAR-expressing therapeutic compositions are described in Section II.C.

[0111] In some embodiments, the disease or condition to be treated is a high-risk multiple myeloma (MM). In some embodiments, the subject has measurable disease as indicated by a serum M-protein level greater than or equal to 0.5 g / dL, as determined by serum protein electrophoresis (SPEP); a urine M- protein level greater than or equal to 200 mg / 24-hour, as determined by urine protein electrophoresis (UPEP); an involved serum free light chain (SFLC) level greater than or equal to 10 mg / dL accompanied by an abnormal kappa / lambda ratio; or any combination of any of the foregoing. In some embodiments, the subject prior to leukapheresis has measurable disease as indicated by a serum M-protein level less than 0.5 g / dL; a urine M-protein level less than 200 mg / 24-hour; and an SFLC level greater than or equal to 10 mg / dL accompanied by an abnormal kappa / lambda ratio.

[0112] In some embodiments, the subject prior to leukapheresis has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1 (see, e.g., Oken et al., (1982) Am J Clin Oncol. 5:649-655). In some embodiments, the Eastern Cooperative Oncology Group (ECOG) performance status indicator can be used to assess or select subjects for treatment, e.g., subjects who have had poor performance from prior therapies (see, e.g., Oken et al., (1982) Am J Clin Oncol.5:649-655). The ECOG Scale of Performance Status describes a patient’s level of functioning in terms of their ability to care for themselves, daily activity, and physical ability (e.g., walking, working, etc.). In some embodiments, an ECOG performance status of 0 indicates that a subject can perform normal activity. In some aspects, subjects with an ECOG performance status of 1 exhibit some restriction in physical activity but the subject is fully ambulatory. In some aspects, patients with an ECOG performance status of 2 is more than 50% ambulatory. In some cases, the subject with an ECOG performance status of 2may also be capable of self-care; see e.g., Sørensen et al., (1993) Br J Cancer 67(4) 773-775. The criteria reflective of the ECOG performance status are described in Table 1 below:

[0113] In some embodiments, prior to, such as at the time of, administration of the provided BCMA-directed CAR T cell compositions, the subject has relapsed following remission after treatment with, or become refractory to, one or more lines of prior therapy for the MM. In any embodiments, at a time prior to leukapheresis in connection with engineering the BCMA-directed CAR T cell composition, the subject has relapsed following remission after treatment with, or become refractory to, one or more lines of prior therapy for treating the MM. Thus, in particular embodiments, prior to the time of treatment, such as prior to leukapheresis, the subject has a R / R MM. In some embodiments, the subject has been previously treated with a therapy or a therapeutic agent targeting the disease or condition, e.g., a MM or a, prior to administration of the cells expressing the recombinant receptor. In some embodiments, the subject has been previously treated with a hematopoietic stem cell transplantation (HSCT), e.g., allogeneic HSCT or autologous HSCT. In some embodiments, the subject has had poor prognosis after treatment with standard therapy and / or has failed one or more lines of previous therapy. In some embodiments, the subject has been treated or has previously received at least or at least about or about 1, 2, 3, 4, or more other therapies for treating the MM, such as a high-risk MM. In some embodiments, the subject has been treated or has previously received a therapy that includes a CD38 targeted agent (e.g. anti-CD38 antibody). In some aspects, the subject has relapsed after an initial response of complete response (CR) or partial response (PR) to the prior therapy. In some embodiments, the subject is refractory to treatment with the at least one or more prior therapy, and the refractory treatment is a best response of stable disease (SD) or progressive disease (PD) after the prior therapy.

[0114] In some embodiments, the subject has relapsed following remission after treatment with, or become refractory to, one or more lines of prior therapy for the disease or condition. In some embodiments, the subject is considered to be refractory to a line of therapy if, among other things, there is documented progressive disease during or within 60 days of completing the last dose of said line of therapy. In some embodiments, the subject has documented progressive disease during or within 60 days of completing the last dose of said line of therapy. In some embodiments, the subject has documentedprogressive disease during or within 12 months of completing the last dose of said line of therapy. In some embodiments, the subject has documented progressive disease during or within the six months prior to administration of the composition of engineered T cells, and the subject is refractory or non-responsive to their most recent line of therapy for treating MM.

[0115] In some embodiments, the subject has relapsed following remission after treatment with, or become refractory to, at least two lines of prior therapy for the disease or condition. In some embodiments, the subject has relapsed following remission after treatment with, or become refractory to, at least three lines of prior therapy for the disease or condition

[0116] In some embodiments, the subject has relapsed following remission after treatment with, or become refractory to, autologous stem cell transplantation (ASCT). In some embodiments, the subject has not received ASCT prior to leukapheresis due to age or other factors.

[0117] In some embodiments, the subject has relapsed following remission after, or become refractory to, at least one cycle of treatment with an immunomodulatory agent. Exemplary immunomodulatory agents include, but are not limited to, thalidomide, lenalidomide, and pomalidomide. In some embodiments, the subject has relapsed following remission after, or become refractory to, at least two consecutive cycles of treatment with an immunomodulatory agent. In some embodiments, the subject has relapsed following remission after, or become refractory to, at least one complete cycle of treatment with an immunomodulatory agent.

[0118] In some embodiments, the immunomodulatory agent is an immune checkpoint inhibitor. In some embodiments, the immunomodulatory agent is an immunomodulatory antibody. Exemplary immune checkpoint inhibitors include Tremelimumab (CTLA-4 blocking antibody, also known as ticilimumab, CP-675,206), anti-OX40, PD-L1 monoclonal antibody (Anti-B7-H1; MEDI4736), MK- 3475 (PD-1 blocker), nivolumab (anti-PD-1 antibody), CT-011 (anti-PD-1 antibody), BY55 monoclonal antibody, AMP224 (anti-PD-L1 antibody), BMS-936559 (anti-PD-L1 antibody), MPLDL3280A (anti- PD-L1 antibody), MSB0010718C (anti-PD-L1 antibody) and ipilimumab (anti-CTLA-4 antibody, also known as Yervoy®, MDX-010 and MDX-101). Exemplary immunomodulatory antibodies include, but are not limited to, Daclizumab (Zenapax), Bevacizumab (Avastin ®), Basiliximab, Ipilimumab, Nivolumab, pembrolizumab, MPDL3280A, Pidilizumab (CT-011), MK-3475, BMS-936559, MPDL3280A (Atezolizumab), tremelimumab, IMP321, BMS-986016, LAG525, urelumab, PF- 05082566, TRX518, MK-4166, dacetuzumab (SGN-40), lucatumumab (HCD122), SEA-CD40, CP-870, CP-893, MEDI6469, MEDI6383, MOXR0916, AMP-224, MSB0010718C (Avelumab), MEDI4736, PDR001, rHIgM12B7, Ulocuplumab, BKT140, Varlilumab (CDX-1127), ARGX-110, MGA271, lirilumab (BMS-986015, IPH2101), IPH2201, ARGX-115, Emactuzumab, CC-90002 and MNRP1685A or an antibody-binding fragment thereof. Other exemplary immunomodulators include, e.g., afutuzumab (available from Roche®); pegfilgrastim (Neulasta®); lenalidomide (CC-5013, Revlimid®); thalidomide(Thalomid®), actimid (CC4047); and IRX-2 (mixture of human cytokines including interleukin 1, interleukin 2, and interferon gamma, CAS 951209-71-5, available from IRX Therapeutics).

[0119] In some embodiments, the subject has relapsed following remission after, or become refractory to, at least one cycle of treatment with a proteasome inhibitor. Exemplary proteasome inhibitors include, but are not limited to, bortezomib, carfilzomib, and ixazomib. In some embodiments, the subject has relapsed following remission after, or become refractory to, at least two consecutive cycles of treatment with a proteasome inhibitor.

[0120] In some embodiments, the subject has relapsed following remission after, or become refractory to, at least two consecutive cycles of treatment with an immunomodulatory agent alone and at least two consecutive cycles of treatment with a proteasome inhibitor alone. In some embodiments, the subject has relapsed following remission after, or become refractory to, at least two consecutive cycles of treatment with an immunomodulatory agent and a proteasome inhibitor in combination.

[0121] In some embodiments, the subject has relapsed following remission after, or become refractory to, treatment with an anti-CD38 antibody. Exemplary anti-CD38 antibodies include, but are not limited to, daratumumab. In some embodiments, the anti-CD38 antibody treatment is a monotherapy. In some embodiments, the anti-CD38 antibody treatment is part of a combination therapy.

[0122] In some embodiments, the subject has relapsed following remission after, or become refractory to, each of (1) ASCT, if eligible to receive ASCT; (2) at least two consecutive cycles of treatment with an immunomodulatory agent alone and at least two consecutive cycles of treatment with a proteasome inhibitor alone; and (3) treatment with an anti-CD38 antibody. In some embodiments, the subject has relapsed following remission after, or become refractory to, each of (1) ASCT, if eligible to receive ASCT; (2) at least two consecutive cycles of treatment with an immunomodulatory agent and a proteasome inhibitor in combination; and (3) treatment with an anti-CD38 antibody.

[0123] In some embodiments, the subject is refractory to the last line of prior therapy administered prior to leukapheresis.

[0124] In some embodiments, the subject prior to leukapheresis does not have active central nervous system (CNS) involvement of MM. In some embodiments, the subject prior to leukapheresis has no history of CNS involvement of MM.

[0125] In some embodiments, the subject prior to leukapheresis does not have active plasma cell leukemia; Waldenstrom’s macroglobulinemia; polyneuropathy, organomegaly, endocrinopathy, monoclonal protein, skin changes (POEMS) syndrome; any clinically significant amyloidosis; or any combination of any of the foregoing. In some embodiments, the subject prior to leukapheresis has no history of plasma cell leukemia; Waldenstrom’s macroglobulinemia; POEMS syndrome; any clinically significant amyloidosis; or any combination of any of the foregoing. In some embodiments, the subject prior to or up to the administration of engineered cells does not have active plasma cell leukemia;Waldenstrom’s macroglobulinemia; POEMS syndrome; any clinically significant amyloidosis; or any combination of any of the foregoing.

[0126] In some embodiments, the subject has not previously received CAR T cell therapy prior to administration of the BCMA-directed engineered CAR T cells in accord with the provided methods. In some embodiments, prior to leukapheresis, the subject has not received genetically-modified T cell therapy. In some embodiments, prior to leukapheresis, the subject has not received BCMA-targeted therapy. Exemplary BCMA-targeted therapies include, but are not limited to, bispecific T cell-engaging antibodies or molecules, antibody-drug conjugates (BCMA-ADC), and BCMA-directed T cell therapy (e.g., BCMA chimeric antigen receptor T cells). In some embodiments, the subject does not have hypersensitivity to fludarabine and / or cyclophosphamide. In some embodiments, the subject does not have an active autoimmune disease requiring immunosuppressive therapy.

[0127] In some embodiments, the subject has not received therapeutic doses of corticosteroids less than 14 days prior to leukapheresis. In some embodiments, a therapeutic dose of corticosteroids is defined as greater than 20 mg / day of prednisone or equivalent. In some embodiments, the subject has not received an anti-MM antibody less than 14 days prior to leukapheresis. In some embodiments, the subject has not received any other approved systemic anti-MM therapy less than 14 days prior to leukapheresis. In some embodiments, the subject has not received any experimental therapy less than 14 days (for biologics) or 5 half-lives (for small molecules) prior to leukapheresis treatment. In some embodiments, the subject has not received an autologous stem-cell transplant (SCT) less than 6 months prior to leukapheresis. In some embodiments, the subject has not received an allogenic SCT less than 6 months prior to leukapheresis. In some embodiments, the subject has not received donor lymphocyte infusions less than 6 weeks prior to leukapheresis. In some embodiments, the subject has not received an immunosuppressive therapy less than 4 weeks prior to leukapheresis. Exemplary immunosuppressive therapies include, but are not limited to, calcineurin inhibitors; methotrexate or other chemotherapeutics; mycophenolate; rapamycin; and immunosuppressive antibodies such as anti-TNF, anti-IL6, or anti-IL6R. In some embodiments, the subject has not undergone plasmapheresis less than 14 days prior to leukapheresis. In some embodiments, the subject has not received radiation therapy targeting an area including a large bone marrow field (e.g. pelvis or sternum) less than 6 weeks prior to leukapheresis. In some embodiments, the subject has not received radiation therapy for a single lesion less than 14 days prior to leukapheresis.

[0128] In some embodiments, the eligibility of subjects for treatment involving administering engineered cells is determined prior to leukapheresis. In some embodiments, the subject prior to leukapheresis has adequate vascular access for leukapheresis. In some embodiments, the subject prior to leukapheresis has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1 (see, e.g., Oken et al., (1982) Am J Clin Oncol.5:649-655). In some embodiments, the subject prior toleukapheresis has recovered, after previous therapy, to less than or equal to Grade 1 from any non- hematological toxicities. In some embodiments, the subject prior to leukapheresis has recovered, after previous therapy, to baseline from any non-hematological toxicities.

[0129] In some embodiments, the subject prior to leukapheresis has adequate organ function. In some embodiments, adequate organ function is indicated by, among other factors, an absolute neutrophil count (ANC) greater than or equal to 1.0 × 109cells / L without growth factor support within 7 days of determination of eligibility; an ANC greater than or equal to 1.0 × 109cells / L without growth factor support within 14 days of determination of eligibility, if pegfilgrastim was previously administered; hemoglobin levels greater than or equal to 8 g / dL without red blood cell (RBC) transfusions within 21 days of determination of eligibility; a platelet count greater than 50 × 109cells / L without transfusion support within 7 days of determination of eligibility; a calculated creatinine clearance rate (serum CrCl, Cockcroft-Gault formula) greater than or equal to 60 mL / min without the support of hydration within 3 days of determination of eligibility; an aspartate aminotransferase (AST) level less than or equal to 3.0 times the upper limit of normal (ULN); an alanine aminotransferase (ALT) level less than or equal to 3.0 times the ULN; a total bilirubin level less than 1.5 times the ULN; a direct bilirubin level less than 1.5 times the ULN, in the case of Gilbert’s syndrome; an international ratio (INR) less than or equal to 1.5 times the ULN; a partial thromboplastin time (PTT) less than or equal to 1.5 times the ULN; adequate pulmonary function, for instance less than or equal to CTCAE Grade 1 dyspnea and / or saturated oxygen (SaO2 greater than 92%) on room air; adequate cardiac function, for instance left ventricular ejection fraction (LVEF) greater than or equal to 40% as assessed by echocardiogram (ECHO) or a multiple uptake gated acquisition (MUGA) scan performed within 8 weeks of determination of eligibility; or a combination of any of the foregoing. Adequate organ function can also be indicated by, among other factors, a calculated creatinine clearance rate (CrCl) greater than or equal to 60 mL / min as measured in 24-hour urine collection without the support of hydration within 3 days of determination of eligibility; and / or a prothrombin time (PT) less than or equal to 1.5 times the ULN.

[0130] In particular embodiments, prior to administration of the dose of BCMA-directed engineered CAR T cells, the subject is administered or has received a lymphodepleting chemotherapy.

[0131] Lymphodepletion may improve the engraftment and activity of CAR T cells through homeostatic cytokines, reduction of CD4+CD25+ regulatory T cells, increase of SDF-1 within bone marrow microenvironment, and stimulatory effects on antigen presenting cells (Grossman et al., Nat Rev Immunol.2004; 4(5):387-395; Stachel et al., Pediatr Blood Cancer 2004; 43(6):644-50; Pinthus et al., J Clin Invest 2004; 114(12):1774-81; Turk et al., J Exp Med 2004; 200(6):771-82). In addition, LD chemotherapy may further reduce the subject’s tumor burden and potentially lower the risk and severity of cytokine release syndrome (CRS).

[0132] Thus, in some embodiments, the methods include administering a preconditioning agent, such as a lymphodepleting or chemotherapeutic agent, such as cyclophosphamide, fludarabine, or combinations thereof, to a subject prior to the administration of engineered cells. For example, the subject may be administered a preconditioning agent at least 2 days prior, such as at least 3, 4, 5, 6, 7, 8, or 9 days prior, to the administration of engineered cells. In some embodiments, the subject is administered a preconditioning agent no more than 9 days prior, such as no more than 8, 7, 6, 5, 4, 3, or 2 days prior, to the administration of engineered cells.

[0133] In some embodiments, the subject is preconditioned with cyclophosphamide at a dose between or between about 20 mg / kg and 100 mg / kg body weight of the subject, such as between or between about 40 mg / kg and 80 mg / kg. In some aspects, the subject is preconditioned or administered with or with about 60 mg / kg of cyclophosphamide. In some embodiments, the cyclophosphamide can be administered in a single dose or can be administered in a plurality of doses, such as given daily, every other day or every three days. In some embodiments, the cyclophosphamide is administered once daily for one or two days. In some embodiments, where the lymphodepleting agent comprises cyclophosphamide, the subject is administered cyclophosphamide at a dose between or between about 100 mg / m2and 500 mg / m2body surface area of the subject, such as between or between about 200 mg / m2and 400 mg / m2, or 250 mg / m2and 350 mg / m2, inclusive. In some instances, the subject is administered about 100 mg / m2of cyclophosphamide. In some instances, the subject is administered about 150 mg / m2of cyclophosphamide. In some instances, the subject is administered about 200 mg / m2of cyclophosphamide. In some instances, the subject is administered about 250 mg / m2of cyclophosphamide. In some instances, the subject is administered about 300 mg / m2of cyclophosphamide. In some embodiments, the cyclophosphamide can be administered in a single dose or can be administered in a plurality of doses, such as given daily, every other day or every three days. In some embodiments, cyclophosphamide is administered daily, such as for 1-5 days, for example, for 3 to 5 days. In some instances, the subject is administered about 300 mg / m2body surface area of the subject, of cyclophosphamide, daily for 3 days, prior to initiation of the cell therapy. In some embodiments, the subject is administered a total of at or about 300 mg / m2, 400 mg / m2, 500 mg / m2, 600 mg / m2, 700 mg / m2, 800 mg / m2, 900 mg / m2, 1000 mg / m2, 1200 mg / m2, 1500 mg / m2, 1800 mg / m2, 2000 mg / m2, 2500 mg / m2, 2700 mg / m2, 3000 mg / m2, 3300 mg / m2, 3600 mg / m2, 4000 mg / m2or 5000 mg / m2cyclophosphamide, or a range defined by any of the foregoing, prior to initiation of the cell therapy.

[0134] In some embodiments, where the lymphodepleting agent comprises fludarabine, the subject is administered fludarabine at a dose between at or about 1 mg / m2and at or 100 mg / m2, such as between at or about 10 mg / m2and at or about 75 mg / m2, at or about 15 mg / m2and at or about 50 mg / m2, at or about 20 mg / m2and at or about 40 mg / m2, at or about or 24 mg / m2and at or about 35 mg / m2, inclusive. In some instances, the subject is administered at or at or about 10 mg / m2of fludarabine. In someinstances, the subject is administered at or about 15 mg / m2of fludarabine. In some instances, the subject is administered at or about 20 mg / m2of fludarabine. In some instances, the subject is administered at or about 25 mg / m2of fludarabine. In some instances, the subject is administered at or about 30 mg / m2of fludarabine. In some embodiments, the fludarabine can be administered in a single dose or can be administered in a plurality of doses, such as given daily, every other day or every three days. In some embodiments, fludarabine is administered daily, such as for 1-5 days, for example, for 3 to 5 days. In some instances, the subject is administered at or about 30 mg / m2body surface area of the subject, of fludarabine, daily for 3 days, prior to initiation of the cell therapy. In some embodiments, the subject is administered a total of at or about 10 mg / m2, 20 mg / m2, 25 mg / m2, 30 mg / m2, 40 mg / m2, 50 mg / m2, 60 mg / m2, 70 mg / m2, 80 mg / m2, 90 mg / m2, 100 mg / m2, 120 mg / m2, 150 mg / m2, 180 mg / m2, 200 mg / m2, 250 mg / m2, 270 mg / m2, 300 mg / m2, 330 mg / m2, 360 mg / m2, 400 mg / m2or 500 mg / m2cyclophosphamide, or a range defined by any of the foregoing, prior to initiation of the cell therapy.

[0135] In some embodiments, the lymphodepleting agent comprises a single agent, such as cyclophosphamide or fludarabine. In some embodiments, the subject is administered cyclophosphamide only, without fludarabine or other lymphodepleting agents. In some embodiments, prior to the administration, the subject has received a lymphodepleting therapy comprising the administration of cyclophosphamide at or about 200-400 mg / m2body surface area of the subject, optionally at or about 300 mg / m2, daily, for 2-4 days. In some embodiments, the subject is administered fludarabine only, for example, without cyclophosphamide or other lymphodepleting agents. In some embodiments, prior to the administration, the subject has received a lymphodepleting therapy comprising the administration of fludarabine at or about 20-40 mg / m2body surface area of the subject, optionally at or about 30 mg / m2, daily, for 2-4 days.

[0136] In some embodiments, the lymphodepleting agent comprises a combination of agents, such as a combination of cyclophosphamide and fludarabine. Thus, the combination of agents may 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 is administered at or about 60 mg / kg (~2 g / m2) of cyclophosphamide and 3 to 5 doses of 25 mg / m2fludarabine prior to the first or subsequent dose. In some the subject is administered fludarabine (30 mg / m2 / day for 3 days) and cyclophosphamide (300 mg / m2 / day for 3 days) (flu / cy) concurrently, intravenously, prior to administration of the cells. In some embodiments, the subject is administered a reduced, delayed or eliminated dose of one or more doses of the lymphodepleting agent(s).

[0137] In some embodiments, after collecting the cells from the subject and prior to administering lymphodepleting (LD) chemotherapy, the subject can receive bridging therapy for disease control. Any of a variety of therapies can be administered as part of a bridging therapy based on the judgment of askilled practitioner for treating the particular disease or condition, including based on factors such as the age of the patient, severity or extent of the disease, potential for side effects, timing of the administration prior to the LD chemotherapy, previous therapies and other factors. In some embodiments, the bridging therapy is administered for no more than four weeks. Exemplary therapies that can be given as a bridge prior to the LD therapy include, but are not limited to, dexamethasone, cyclophosphamide, etoposide, and cisplatin (DCEP); bortezomib, dexamethasone, cisplatin, doxorubicin, cyclophosphamide, and etoposide (VD-PACE); cyclophosphamide, vincristine, doxorubicin, and dexamethasone (CVAD); pulsed dexamethasone; and an approved daratumumab-containing regimen. In some embodiments, the bridging therapy is discontinued at least 14 days prior to LD therapy.. In some embodiments, bridging therapies are discontinued 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. In some embodiments, subjects must recover to Grade 2 or lower from bridging therapy-related toxicities prior to LD chemotherapy.

[0138] In some embodiments, the subjects are premedicated, e.g., to minimize the risk of infusion reaction. In some aspects, the premedication includes administering pain reliever and / or an antihistamine. In some embodiments, the premedication includes administering an acetaminophen and / or a diphenhydramine, or another H1-antihistamine. In some embodiments, the patient with acetaminophen (e.g., 650 mg orally) and diphenhydramine (e.g., 25-50 mg, IV or orally), or another H1-antihistamine, at or about 30 to 60 minutes prior to treatment with the cell therapy.

[0139] In some embodiments, the subject is at least 18 years of age. In embodiments of any of the provided methods, the subject is a human subject. A. Dosing

[0140] In some embodiments, a dose of engineered cells is administered to subjects in accordance with the provided methods, and / or with the provided articles of manufacture or compositions. In some embodiments, the size or timing of the doses is determined as a function of the particular disease or condition in the subject. In some cases, the size or timing of the doses for a particular disease in view of the provided description may be empirically determined.

[0141] In some embodiments, the treatment does not induce an immune response by the subject to the therapy, and / or does not induce such a response to a degree that prevents effective treatment of the disease or condition. In some aspects, the degree of immunogenicity and / or graft versus host response is less than that observed with a different but comparable treatment. For example, in the case of adoptive cell therapy using cells expressing CARs including the provided anti-BCMA antibodies, the degree of immunogenicity in some embodiments is reduced compared to CARs including a different antibody that binds to a similar, e.g., overlapping epitope and / or that competes for binding to BCMA with the antibody, such as a mouse or monkey or rabbit or humanized antibody.

[0142] In some embodiments, the methods include adoptive cell therapy, whereby genetically engineered cells expressing the provided recombinant receptors comprising a BCMA-binding molecule (e.g., CARs comprising anti-BCMA antibody or antigen-binding fragment thereof) are administered to subjects. Such administration can promote activation of the cells (e.g., T cell activation) in a BCMA- targeted manner, such that the cells of the disease or disorder are targeted for destruction.

[0143] Thus, the provided methods and uses include methods and uses for adoptive cell therapy. In some embodiments, the methods include administration of the cells or a composition containing the cells to a subject, tissue, or cell, such as one having, at risk for, or suspected of having the disease, condition or disorder. In some embodiments, the cells, populations, and compositions are administered to a subject having the particular disease or condition to be treated, e.g., via adoptive cell therapy, such as adoptive T cell therapy. In some embodiments, the cells or compositions are administered to the subject, such as a subject having or at risk for the disease or condition. In some aspects, the methods thereby treat, e.g., ameliorate one or more symptom of the disease or condition, such as by lessening tumor burden in a BCMA-expressing cancer.

[0144] Methods for administration of cells for adoptive cell therapy are known and may be used in connection with the provided methods and compositions. For example, adoptive T cell therapy methods are described, e.g., in US Patent Application Publication No.2003 / 0170238 to Gruenberg et al; US Patent No.4,690,915 to Rosenberg; Rosenberg (2011) Nat Rev Clin Oncol.8(10):577-85). See, e.g., 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.

[0145] In some embodiments, the cell therapy, e.g., adoptive cell therapy, e.g., adoptive T cell therapy, is carried out by autologous transfer, in which the cells are isolated and / or otherwise prepared from the subject who is to receive the cell therapy, or from a sample derived from such a subject. Thus, in some aspects, the cells are derived from a subject, e.g., patient, in need of a treatment and the cells, following isolation and processing are administered to the same subject.

[0146] In some embodiments, the cell therapy, e.g., adoptive cell therapy, e.g., adoptive T cell therapy, is carried out by allogeneic transfer, in which the cells are isolated and / or otherwise prepared from a subject other than a subject who is to receive or who ultimately receives the cell therapy, e.g., a first subject. In such embodiments, the cells then are administered to a different subject, e.g., a second subject, of the same species. 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.

[0147] In some embodiments, the subject, to whom the cells, cell populations, or compositions are administered, is a primate, such as a human. In some embodiments, the subject, to whom the cells, cell populations, or compositions are administered, is a non-human primate. In some embodiments, the non-human primate is a monkey (e.g., cynomolgus monkey) or an ape. The subject can be male or female and can be any suitable age, including infant, juvenile, adolescent, adult, and geriatric subjects. In some embodiments, the subject is a non-primate mammal, such as a rodent (e.g., mouse, rat, etc.). In some examples, the patient or subject is a validated animal model for disease, adoptive cell therapy, and / or for assessing toxic outcomes such as cytokine release syndrome (CRS).

[0148] The BCMA-binding molecules such as recombinant receptors (e.g., CARs) and cells expressing the same, can be administered by any suitable means, for example, by injection, e.g., intravenous or subcutaneous injections, intraocular injection, periocular injection, subretinal injection, intravitreal injection, trans-septal injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjunctival injection, subconjunctival injection, sub-Tenon’s injection, retrobulbar injection, peribulbar injection, or posterior juxtascleral delivery. In some embodiments, they are administered by parenteral, intrapulmonary, and intranasal, and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, intracranial, intrathoracic, or subcutaneous administration. Dosing and administration may depend in part on whether the administration is brief or chronic. Various dosing schedules include but are not limited to single or multiple administrations over various time-points, bolus administration, and pulse infusion.

[0149] For the prevention or treatment of disease, the appropriate dosage of the binding molecule, recombinant receptor or cell may depend on the type of disease to be treated, the type of binding molecule or recombinant receptor, the severity and course of the disease, whether the binding molecule or recombinant receptor is administered for preventive or therapeutic purposes, previous therapy, the patient’s clinical history and response to the recombinant receptor or cell, and the discretion of the attending physician. The compositions and molecules and cells are in some embodiments suitably administered to the patient at one time or over a series of treatments.

[0150] In some embodiments, the methods comprises administering a dose of the engineered cells or a composition comprising a dose of the engineered cells. In some embodiments, the engineered cells or compositions containing engineered cells can be used in a treatment regimen, wherein the treatment regimen comprises administering a dose of the engineered cells or a composition comprising a dose of the engineered cells. In some embodiments, the dose can contain, for example, a particular number or range of recombinant receptor-expressing T cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), such as any number of such cells described herein. In some embodiments, a composition containing a dose of the cells can be administered. In some aspects, the number, amount or proportion of CAR-expressing (CAR+) cells in a cell population or a cell composition can be assessed by detection of a surrogate marker, e.g., by flow cytometry or other means, or by detecting binding of a labelled molecule,such as a labelled antigen, that can specifically bind to the binding molecules or receptors provided herein.

[0151] In some of any of the provided embodiments, the dose of T cells, such as engineered T cells expressing the BCMA-directed CAR, includes is enriched for, or comprises a cell composition or a cell population that is enriched for, CD3+ T cells, CD4+ T cells, CD8+ T cells or CD4+ T cells and CD8+ T cells. In some of any such embodiments, greater than at or about 70%, 75%, 80%, 85%, 90%, 95% or 98% of the cells in the dose of T cells are CD3+ T cells, CD4+ T cells, CD8+ T cells or CD4+ T cells and CD8+ T cells. In some of any such embodiments, greater than at or about 70%, 75%, 80%, 85%, 90%, 95% or 98% of the cells in the dose of T cells are CD3+ T cells. In some of any of the provided embodiments, the dose of T cells comprises both CD4+ cells and CD8+ cells. In some of any such embodiments, greater than at or about 70%, 75%, 80%, 85%, 90%, 95% or 98% of the cells in the dose of T cells are CD4+ T cells and CD8+ T cells.

[0152] In some embodiments, the dose of cells comprises between at or about 0.1 x 105of the BCMA-directed CAR engineered cells per kilogram body weight of the subject (cells / kg) and at or about 2 x 106cells / kg, such as between at or about 0.1 x 105cells / kg and at or about 0.5 x 105cells / kg, between at or about 0.5 x 105cells / kg and at or about 1 x 105cells / kg, between at or about 1 x 105cells / kg and at or about 1.5 x 105cells / kg, between at or about 1.5 x 105cells / kg and at or about 2 x 105cells / kg, between at or about 2 x 105cells / kg and at or about 2.5 x 105cells / kg, between at or about 2.5 x 105cells / kg and at or about 3 x 105cells / kg, between at or about 3 x 105cells / kg and at or about 3.5 x 105cells / kg, between at or about 3.5 x 105cells / kg and at or about 4 x 105cells / kg, between at or about 4 x 105cells / kg and at or about 4.5 x 105cells / kg, between at or about 4.5 x 105cells / kg and at or about 5 x 105cells / kg, between at or about 5 x 105cells / kg and at or about 5.5 x 105cells / kg, between at or about 5.5 x 105cells / kg and at or about 6 x 105cells / kg, between at or about 6 x 105cells / kg and at or about 6.5 x 105cells / kg, between at or about 6.5 x 105cells / kg and at or about 7 x 105cells / kg, between at or about 7 x 105cells / kg and at or about 7.5 x 105cells / kg, between at or about 7.5 x 105cells / kg and at or about 8 x 105cells / kg, or between at or about 8 x 105of the cells / kg and at or about 10 x 105of the cells / kg. In some embodiments, the dose of cells comprises no more than 2 x 105of the BCMA -directed CAR engineered cells per kilogram body weight of the subject (cells / kg), such as no more than at or about 3 x 105cells / kg, no more than at or about 4 x 105cells / kg, no more than at or about 5 x 105cells / kg, no more than at or about 6 x 105cells / kg, no more than at or about 7 x 105cells / kg, no more than at or about 8 x 105cells / kg, no more than at or about 9 x 105cells / kg, no more than at or about 1 x 106cells / kg, or no more than at or about 2 x 106cells / kg. In some embodiments, the dose of cells comprises at least or at least about or at or about 0.1 x 105of the BCMA-directed CAR engineered cells per kilogram body weight of the subject (cells / kg), such as at least or at least about or at or about 0.2 x 105cells / kg, at least or at least about or at or about 0.3 x 105cells / kg, at least or at least about or at or about 0.4 x 105cells / kg,at least or at least about or at or about 0.5 x 105cells / kg, at least or at least about or at or about 0.6 x 105cells / kg, at least or at least about or at or about 0.7 x 105cells / kg, at least or at least about or at or about 0.8 x 105cells / kg, at least or at least about or at or about 0.9 x 105cells / kg, at least or at least about or at or about 0.1 x 106cells / kg, or at least or at least about or at or about 0.2 x 106cells / kg. In some embodiments, the number of cells is the number of such cells that are viable cells, e.g., viable T cells such as viable CD3+ cells expressing the BCMA-directed CAR.

[0153] In certain embodiments, the cells, or individual populations of sub-types of cells, are administered to the subject at a range of at or about 0.1 million to at or about 100 billion cells and / or that amount of cells per kilogram of body weight of the subject, such as, e.g., at or about 0.1 million to at or about 50 billion cells (e.g., at or about 5 million cells, at or about 25 million cells, at or about 500 million cells, at or about 1 billion cells, at or about 5 billion cells, at or about 20 billion cells, at or about 30 billion cells, at or about 40 billion cells, or a range defined by any two of the foregoing values), at or about 1 million to at or about 50 billion cells (e.g., at or about 5 million cells, at or about 25 million cells, at or about 500 million cells, at or about 1 billion cells, at or about 5 billion cells, at or about 20 billion cells, at or about 30 billion cells, at or about 40 billion cells, or a range defined by any two of the foregoing values), such as at or about 10 million to at or about 100 billion cells (e.g., at or about 20 million cells, at or about 30 million cells, at or about 40 million cells, at or about 60 million cells, at or about 70 million cells, at or about 80 million cells, at or about 90 million cells, at or about 10 billion cells, at or about 25 billion cells, at or about 50 billion cells, at or about 75 billion cells, at or about 90 billion cells, or a range defined by any two of the foregoing values), and in some cases at or about 100 million cells to at or about 50 billion cells (e.g., at or about 120 million cells, at or about 250 million cells, at or about 350 million cells, at or about 650 million cells, at or about 800 million cells, at or about 900 million cells, at or about 3 billion cells, at or about 30 billion cells, at or about 45 billion cells) or any value in between these ranges and / or per kilogram of body weight of the subject. Dosages may vary depending on attributes particular to the disease or disorder and / or patient and / or other treatments. In some embodiments, such values refer to numbers of recombinant receptor-expressing cells; in other embodiments, they refer to number of T cells or total cells in the composition administered. In some embodiments, the number of cells is the number of such cells that are viable cells.

[0154] In some embodiments, the dose of cells is a flat dose of cells or fixed dose of cells such that the dose of cells is not tied to or based on the body surface area or weight of a subject.

[0155] In some embodiments, the dose of genetically engineered cells comprises from at or about 1 x 105to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 1.6 x108total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 1.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 1.5 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 1.5 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 1.5 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 1.5 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 1.5 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 1.5 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 1.5 x 107to at or about 1.0 x 108total T cellsexpressing the BCMA-directed CAR, from at or about 2 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 2 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 2 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 2 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 2 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 2 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 2 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 2 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 2.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 2.5 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 2.5 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 2.5 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 2.5 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 2.5 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 2.5 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 2.5 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 3 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 3 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 3 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 3 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 3 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 3 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 3 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 3 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 3.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 3.5 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 3.5 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 3.5 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 3.5 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 3.5 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 3.5 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 3.5 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 4 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 4 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 4 x 107to at or about 2.0 x 108total T cellsexpressing the BCMA-directed CAR, from at or about 4 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 4 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 4 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 4 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 4 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 4.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 4.5 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 4.5 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 4.5 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 4.5 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 4.5 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 4.5 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 4.5 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 5.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 5.5 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 5.5 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 5.5 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 5.5 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 5.5 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 5.5 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 5.5 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 6 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 6 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 6 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 6 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 6 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 6 x 107to at or about 1.4 x 108total T cellsexpressing the BCMA-directed CAR, from at or about 6 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 6 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 6.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 6.5 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 6.5 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 6.5 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 6.5 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 6.5 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 6.5 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 6.5 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 7 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 7 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 7 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 7 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 7 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 7 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 7 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 7 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 7.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 7.5 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 7.5 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 7.5 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 7.5 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 7.5 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 7.5 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 7.5 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 8 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 8 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 8 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 8 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 8 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 8 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 8 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 8 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 8.5 x 107to at or about 2.4 x 108total T cellsexpressing the BCMA-directed CAR, from at or about 8.5 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 8.5 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 8.5 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 8.5 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 8.5 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 8.5 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 8.5 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 9 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 9 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 9 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 9 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 9 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 9 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 9 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 9 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 9.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 9.5 x 107to at or about 2.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 9.5 x 107to at or about 2.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 9.5 x 107to at or about 1.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 9.5 x 107to at or about 1.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 9.5 x 107to at or about 1.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 9.5 x 107to at or about 1.2 x 108total T cells expressing the BCMA-directed CAR, or from at or about 9.5 x 107to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the number of cells is the number of such cells that are viable cells, such as viable T cells. In some embodiments, the number of cells is the number of such cells that are CD3+ cells. In some embodiments, the number of cells is the number of such cells that are CD4+ or CD8+ cells.

[0156] In some embodiments, the dose of genetically engineered cells comprises from at or about 1 x 105to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 5 x 105to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 1 x 106to at or about 2.4 x 108total T cells expressing the BCMA- directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 5 x 106to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 1 x 107to at or about 2.4 x 108total Tcells expressing the BCMA-directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 1.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA- directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 2 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 2.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 3 x 107to at or about 2.4 x 108total T cells expressing the BCMA- directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 3.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 4.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 5.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 6 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 6.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 7 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 7.5 x 107to at or about 2.4 x 108total T cells expressing the BCMA- directed CAR. In some embodiments, the dose of genetically engineered cells comprises from at or about 8 x 107to at or about 2.4 x 108total T cells expressing the BCMA-directed CAR. In some embodiments, the number of cells is the number of such cells that are viable cells, such as viable T cells. In some embodiments, the number of cells is the number of such cells that are CD3+ cells. In some embodiments, the number of cells is the number of such cells that are CD4+ or CD8+ cells.

[0157] In some embodiments, the dose of genetically engineered cells comprises from at or about 1 x 105to at or about 1 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 0.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 0.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 0.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 0.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 1.0 x 107total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 0.8 x 107total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 0.6 x 107total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 0.4 x 107total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 0.2 x 107total T cells expressing the BCMA-directed CAR, from at or about 1 x 105to at or about 1.0 x 106total T cellsexpressing the BCMA-directed CAR, from at or about 1 x 106to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 0.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 0.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 0.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 0.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 1.0 x 107total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 0.8 x 107total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 0.6 x 107total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 0.4 x 107total T cells expressing the BCMA-directed CAR, from at or about 1 x 106to at or about 0.2 x 107total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 0.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 0.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 0.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 0.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 1.0 x 107total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 0.8 x 107total T cells expressing the BCMA-directed CAR, from at or about 5 x 106to at or about 0.6 x 107total T cells expressing the BCMA-directed CAR, from at or about 10 x 106to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 10 x 106to at or about 0.9 x 108total T cells expressing the BCMA-directed CAR, from at or about 10 x 106to at or about 0.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 10 x 106to at or about 0.7 x 108total T cells expressing the BCMA-directed CAR, from at or about 10 x 106to at or about 0.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 10 x 106to at or about 0.5 x 108total T cells expressing the BCMA-directed CAR, from at or about 10 x 106to at or about 0.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 10 x 106to at or about 0.3 x 108total T cells expressing the BCMA-directed CAR, from at or about 10 x 106to at or about 0.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 10 x 106to at or about 15 x 106total T cells expressing the BCMA-directed CAR, from at or about 15 x 106to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 15 x 106to at or about 0.9 x 108total T cells expressing the BCMA-directed CAR, from at or about 15 x 106to at or about 0.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 15 x 106to at or about 0.7 x 108total T cells expressing the BCMA-directed CAR, from at or about 15 x 106to at or about 0.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 15 x 106to at or about 0.5 x 108total T cells expressing the BCMA-directed CAR, from at or about 15 x 106to at or about 0.4 x 108total T cellsexpressing the BCMA-directed CAR, from at or about 15 x 106to at or about 0.3 x 108total T cells expressing the BCMA-directed CAR, from at or about 15 x 106to at or about 0.2 x 108total T cells expressing the BCMA-directed CAR, from at or about 20 x 106to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 20 x 106to at or about 0.9 x 108total T cells expressing the BCMA-directed CAR, from at or about 20 x 106to at or about 0.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 20 x 106to at or about 0.7 x 108total T cells expressing the BCMA-directed CAR, from at or about 20 x 106to at or about 0.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 20 x 106to at or about 0.5 x 108total T cells expressing the BCMA-directed CAR, from at or about 20 x 106to at or about 0.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 20 x 106to at or about 0.3 x 108total T cells expressing the BCMA-directed CAR, from at or about 20 x 106to at or about 25 x 106total T cells expressing the BCMA-directed CAR, from at or about 25 x 106to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 25 x 106to at or about 0.9 x 108total T cells expressing the BCMA-directed CAR, from at or about 25 x 106to at or about 0.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 25 x 106to at or about 0.7 x 108total T cells expressing the BCMA-directed CAR, from at or about 25 x 106to at or about 0.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 25 x 106to at or about 0.5 x 108total T cells expressing the BCMA-directed CAR, from at or about 25 x 106to at or about 0.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 25 x 106to at or about 0.3 x 108total T cells expressing the BCMA-directed CAR, from at or about 30 x 106to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 30 x 106to at or about 0.9 x 108total T cells expressing the BCMA-directed CAR, from at or about 30 x 106to at or about 0.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 30 x 106to at or about 0.7 x 108total T cells expressing the BCMA-directed CAR, from at or about 30 x 106to at or about 0.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 30 x 106to at or about 0.5 x 108total T cells expressing the BCMA-directed CAR, from at or about 30 x 106to at or about 0.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 30 x 106to at or about 35 x 106total T cells expressing the BCMA-directed CAR, from at or about 35 x 106to at or about 1.0 x 108total T cells expressing the BCMA-directed CAR, from at or about 35 x 106to at or about 0.9 x 108total T cells expressing the BCMA-directed CAR, from at or about 35 x 106to at or about 0.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 35 x 106to at or about 0.7 x 108total T cells expressing the BCMA-directed CAR, from at or about 35 x 106to at or about 0.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 35 x 106to at or about 0.5 x 108total T cells expressing the BCMA-directed CAR, from at or about 35 x 106to at or about 0.4 x 108total T cells expressing the BCMA-directed CAR, from at or about 40 x 106to at or about 1.0 x 108total T cellsexpressing the BCMA-directed CAR, from at or about 40 x 106to at or about 0.9 x 108total T cells expressing the BCMA-directed CAR, from at or about 40 x 106to at or about 0.8 x 108total T cells expressing the BCMA-directed CAR, from at or about 40 x 106to at or about 0.7 x 108total T cells expressing the BCMA-directed CAR, from at or about 40 x 106to at or about 0.6 x 108total T cells expressing the BCMA-directed CAR, from at or about 40 x 106to at or about 0.5 x 108total T cells expressing the BCMA-directed CAR, or from at or about 40 x 106to at or about 45 x 106total T cells expressing the BCMA-directed CAR. In some embodiments, the number of cells is the number of such cells that are viable cells, such as viable T cells.

[0158] In some embodiments, the dose of genetically engineered cells comprises at least or at least about 1 x 105T cells expressing the BCMA-directed CAR, at least or at least about 2.5 x 105T cells expressing the BCMA-directed CAR, at least or at least about 5 x 105T cells expressing the BCMA- directed CAR, at least or at least about 1 x 106T cells expressing the BCMA-directed CAR, at least or at least about 2.5 x 106T cells expressing the BCMA-directed CAR, at least or at least about 5 x 106T cells expressing the BCMA-directed CAR, at least or at least about 1 x 107T cells expressing the BCMA- directed CAR, at least or at least about 2.5 x 107T cells expressing the BCMA-directed CAR, or at least or at least about 5 x 107T cells expressing the BCMA-directed CAR. In some embodiments, the number of cells is the number of such cells that are viable cells, such as viable T cells.

[0159] In some embodiments, the dose of genetically engineered cells comprises less than or less than about 1 x 105T cells expressing the BCMA-directed CAR, less than or less than about 2.5 x 105T cells expressing the BCMA-directed CAR, less than or less than about 5 x 105T cells expressing the BCMA-directed CAR, less than or less than about 1 x 106T cells expressing the BCMA-directed CAR, less than or less than about 2.5 x 106T cells expressing the BCMA-directed CAR, less than or less than about 5 x 106T cells expressing the BCMA-directed CAR, less than or less than about 1 x 107T cells expressing the BCMA-directed CAR, less than or less than about 1.5 x 107T cells expressing the BCMA-directed CAR, less than or less than about 2 x 107T cells expressing the BCMA-directed CAR, less than or less than about 2.5 x 107T cells expressing the BCMA-directed CAR, less than or less than about 3 x 107T cells expressing the BCMA-directed CAR, less than or less than about 3.5 x 107T cells expressing the BCMA-directed CAR, less than or less than about 4 x 107T cells expressing the BCMA- directed CAR, less than or less than about 4.5 x 107T cells expressing the BCMA-directed CAR, or less than or less than about 5 x 107T cells expressing the BCMA-directed CAR. In some embodiments, the number of cells is the number of such cells that are viable cells, such as viable T cells.

[0160] In some embodiments, the cell therapy comprises administration of a dose comprising a number of cell from or from about 1 x 105to or to about 1 x 108total recombinant receptor-expressing cells or total T cells, from or from about 5 x 105to or to about 5 x 107total recombinant receptor- expressing cells or total T cells, or from or from about 1 x 106to or to about 1 x 107total recombinantreceptor-expressing cells or total T cells,, each inclusive. In some embodiments, the cell therapy comprises administration of a dose comprising a number of cell from or from about 1 x 105to or to about 1 x 108total recombinant receptor-expressing cells, or total T cells, from or from about 5 x 105to or to about 1 x 108total recombinant receptor-expressing cells, or total T cells, from or from about 1 x 106to or to about 50 x 106total recombinant receptor-expressing cells, or total T cells, from or from about 5 x 106to or to about 45 x 106total recombinant receptor-expressing cells or total T cells, or from or from about 10 x 106to or to about 25 x 106total recombinant receptor-expressing cells or total T cells, each inclusive. In some embodiments, the cell therapy comprises administration of a dose of cells comprising a number of cells at least or at least about 1 x 105total recombinant receptor-expressing cells or total T cells, such at least or at least 1 x 106, at least or at least about 1 x 107, at least or at least about 1 x 108of such cells. In some embodiments, the number of cells is the number of such cells that are viable cells, such as viable T cells.

[0161] In some embodiments, for example, where the subject is a human, the dose includes more than at or about 5 × 106total CAR-expressing (CAR+) cells, T cells, or peripheral blood mononuclear cells (PBMCs) and fewer than at or about 100 × 106total CAR-expressing cells, T cells, or PBMCs. In some embodiments, the dose of genetically engineered cells comprises from at or about 5 × 106to at or about 10 × 106total CAR-expressing (CAR+) T cells, from at or about 10 × 106to at or about 15 × 106CAR+ T cells, from at or about 15 × 106to at or about 20 × 106CAR+ T cells, from at or about 20 × 106to at or about 25 × 106CAR+ T cells, from at or about 25 × 106to at or about 30 × 106CAR+ T cells, from at or about 30 × 106to at or about 35 × 106CAR+ T cells, from at or about 35 × 106to at or about 40 × 106CAR+ T cells, from at or about 40 × 106to at or about 45 × 106CAR+ T cells, from at or about 45 × 106to at or about 50 × 106CAR+ T cells, from at or about 50 × 106to at or about 55 × 106CAR+ T cells, from at or about 55 × 106to at or about 60 × 106CAR+ T cells, from at or about 60 × 106to at or about 65 × 106CAR+ T cells, from at or about 65 × 106to at or about 70 × 106CAR+ T cells, from at or about 70 × 106to at or about 75 × 106CAR+ T cells, from at or about 75 × 106to at or about 80 × 106CAR+ T cells, from at or about 80 × 106to at or about 85 × 106CAR+ T cells, from at or about 85 × 106to at or about 90 × 106CAR+ T cells, from at or about 90 × 106to at or about 95 × 106CAR+ T cells, or from at or about 95 × 106to at or about 100 × 106CAR+ T cells, each inclusive. In any of the preceding embodiments, the CAR+ T cells express a BCMA-targeting CAR such as one derived from BCMA-55.

[0162] In some embodiments, for example, where the subject is a human, the dose includes at or about 5 × 106total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). In some embodiments, for example, where the subject is a human, the dose includes at or about 10 × 106total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). In some embodiments, for example, where the subject is a human, the dose includes at or about 20 × 106total recombinant receptor (e.g., CAR)-expressing cells, Tcells, or peripheral blood mononuclear cells (PBMCs). In some embodiments, for example, where the subject is a human, the dose includes at or about 30 × 106total recombinant receptor (e.g., CAR)- expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). In some embodiments, for example, where the subject is a human, the dose includes at or about 40 × 106total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). In some embodiments, for example, where the subject is a human, the dose includes at or about 60 × 106total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). In some embodiments, for example, where the subject is a human, the dose includes at or about 80 × 106total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs).

[0163] In some embodiments, the number is with reference to the total number of CD3+ or CD8+, in some cases also CAR-expressing (e.g. CAR+) cells. In some embodiments, the dose of genetically engineered cells comprises from at or about 1 x 107to at or about 1.5 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, from at or about 1.5 x 107to at or about 2 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, from at or about 2 x 107to at or about 2.5 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, from at or about 2.5 x 107to at or about 3 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, from at or about 3 x 107to at or about 3.5 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, from at or about 3.5 x 107to at or about 4 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, from at or about 4 x 107to at or about 4.5 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, from at or about 4.5 x 107to at or about 5 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, from at or about 5 x 107to at or about 5.5 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, from at or about 5.5 x 107to at or about 6 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, from at or about 6 x 107to at or about 6.5 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, from at or about 6.5 x 107to at or about 7.5 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, or from at or about 7.5 x 107to at or about 8 x 107CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR- expressing cells, each inclusive.

[0164] In some embodiments, the dose of genetically engineered cells is with reference to the total number of CD3+ CAR-expressing (CAR+) or CD4+ / CD8+ CAR-expressing (CAR+) cells. In some embodiments, the dose of genetically engineered cells comprises from at or about 1 x 107to at or about 1.5 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, from at or about 1.5 x 107to at or about 2 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR- expressing cells, from at or about 2 x 107to at or about 2.5 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, from at or about 2.5 x 107to at or about 3 x 107CD3+ orCD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, from at or about 3 x 107to at or about 3.5 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, from at or about 3.5 x 107to at or about 4 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, from at or about 4 x 107to at or about 4.5 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, from at or about 4.5 x 107to at or about 5 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, from at or about 5 x 107to at or about 5.5 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, from at or about 5.5 x 107to at or about 6 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, from at or about 6 x 107to at or about 6.5 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, from at or about 6.5 x 107to at or about 7.5 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR- expressing cells, or from at or about 7.5 x 107to at or about 8 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, each inclusive.

[0165] In some embodiments, the dose comprises at or about 1.0 x 107, 2.0 x 107, 3.0 x 107, 4.0 x 107, 6.0 x 107, or 8.0 x 107CD3+ or CD4+ / CD8+ total T cells or CD3+ CAR-expressing or CD4+ / CD8+ CAR-expressing cells. In some embodiments, the dose comprises at or about 1.0 x 107, 2.0 x 107, 3.0 x 107, 4.0 x 107, 6.0 x 107, or 8.0 x 107CD3+ CAR-expressing cells. In some embodiments, the dose comprises at or about 1.0 x 107, 2.0 x 107, 3.0 x 107, 4.0 x 107, 6.0 x 107, or 8.0 x 107CD4+ / CD8+ CAR- expressing cells.

[0166] In some embodiments, the dose is between at or about 0.5 x 107CD3+ CAR-expressing cells and at or about 1.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 1.5 x 107CD3+ CAR-expressing cells and at or about 2.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 2.5 x 107CD3+ CAR-expressing cells and at or about 3.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 3.5 x 107CD3+ CAR-expressing cells and at or about 4.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 4.5 x 107CD3+ CAR-expressing cells and at or about 5.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 5.5 x 107CD3+ CAR-expressing cells and at or about 6.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 6.5 x 107CD3+ CAR-expressing cells and at or about 7.5 x 107CD3+ CAR- expressing cells. In some embodiments, the dose is between at or about 7.5 x 107CD3+ CAR-expressing cells and at or about 8.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 8.5 x 107CD3+ CAR-expressing cells and at or about 9.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 9.5 x 107CD3+ CAR-expressing cells and at or about 10.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 10.5 x 107CD3+ CAR-expressing cells and at or about 11.5 x 107CD3+ CAR-expressing cells. In someembodiments, the dose is between at or about 11.5 x 107CD3+ CAR-expressing cells and at or about 12.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 12.5 x 107CD3+ CAR-expressing cells and at or about 13.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 13.5 x 107CD3+ CAR-expressing cells and at or about 14.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 14.5 x 107CD3+ CAR-expressing cells and at or about 15.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 15.5 x 107CD3+ CAR-expressing cells and at or about 16.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 16.5 x 107CD3+ CAR-expressing cells and at or about 17.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 17.5 x 107CD3+ CAR-expressing cells and at or about 18.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 18.5 x 107CD3+ CAR-expressing cells and at or about 19.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 19.5 x 107CD3+ CAR-expressing cells and at or about 20.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 20.5 x 107CD3+ CAR-expressing cells and at or about 21.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 21.5 x 107CD3+ CAR-expressing cells and at or about 22.5 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is between at or about 22.5 x 107CD3+ CAR-expressing cells and at or about 23.5 x 107CD3+ CAR-expressing cells.

[0167] In some embodiments, the dose is at or about 1.0 x 107CD3+ CAR-expressing cells, at or about 2.0 x 107CD3+ CAR-expressing cells, at or about 3.0 x 107CD3+ CAR-expressing cells, at or about 4.0 x 107CD3+ CAR-expressing cells, at or about 5.0 x 107CD3+ CAR-expressing cells, at or about 6.0 x 107CD3+ CAR-expressing cells, at or about 7.0 x 107CD3+ CAR-expressing cells, at or about 8.0 x 107CD3+ CAR-expressing cells, at or about 9.0 x 107CD3+ CAR-expressing cells, at or about 10.0 x 107CD3+ CAR-expressing cells, at or about 11.0 x 107CD3+ CAR-expressing cells, at or about 12.0 x 107CD3+ CAR-expressing cells, at or about 13.0 x 107CD3+ CAR-expressing cells, at or about 14.0 x 107CD3+ CAR-expressing cells, at or about 15.0 x 107CD3+ CAR-expressing cells, at or about 16.0 x 107CD3+ CAR-expressing cells, at or about 17.0 x 107CD3+ CAR-expressing cells, at or about 18.0 x 107CD3+ CAR-expressing cells, at or about 19.0 x 107CD3+ CAR-expressing cells, at or about 20.0 x 107CD3+ CAR-expressing cells, at or about 21.0 x 107CD3+ CAR-expressing cells, at or about 22.0 x 107CD3+ CAR-expressing cells, at or about 23.0 x 107CD3+ CAR-expressing cells, or at or about 24.0 x 107CD3+ CAR-expressing cells.

[0168] In some embodiments, the dose is at or about 1.0 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is at or about 2.0 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is at or about 3.0 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is at or about 4.0 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is at or about 6.0 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is at or about 8.0 x 108CD3+ CAR- expressing cells.

[0169] In some embodiments, the dose is at or about 8.0 x 107CD3+ CAR-expressing cells. In some embodiments, the dose is at or about 16.0 x 107CD3+ CAR-expressing cells.

[0170] In some embodiments, the dose is at or about 1.0 x 107CD4+ / CD8+ CAR-expressing cells. In some embodiments, the dose is at or about 2.0 x 107CD4+ / CD8+ CAR-expressing cells. In some embodiments, the dose is at or about 3.0 x 107CD4+ / CD8+ CAR-expressing cells. In some embodiments, the dose is at or about 4.0 x 107CD4+ / CD8+ CAR-expressing cells. In some embodiments, the dose is at or about 6.0 x 107CD4+ / CD8+ CAR-expressing cells. In some embodiments, the dose is at or about 8.0 x 108CD4+ / CD8+ CAR-expressing cells.

[0171] In some embodiments, the dose is at or about 8.0 x 107CD4+ / CD8+ CAR-expressing cells. In some embodiments, the dose is at or about 16.0 x 107CD4+ / CD8+ CAR-expressing cells.

[0172] In some embodiments, the dose of cells, e.g., recombinant receptor-expressing T cells, is administered to the subject as a single dose or is administered only one time within a period of two weeks, one month, three months, six months, 1 year or more. In some embodiments, the patient is administered multiple doses, and each of the doses or the total dose can be within any of the foregoing values.

[0173] In some embodiments, the engineered cells for administration or composition of engineered cells for administration, exhibits properties indicative of or consistent with cell health. In some embodiments, at or about or at least at or about 70, 75, 80, 85, or 90% CAR+ cells of such dose exhibit one or more properties or phenotypes indicative of cell health or biologically active CAR cell, such as absence expression of an apoptotic marker.

[0174] In particular embodiments, the phenotype is or includes an absence of apoptosis and / or an indication the cell is undergoing the apoptotic process. Apoptosis is a process of programmed cell death that includes a series of stereotyped morphological and biochemical events that lead to characteristic cell changes and death, including blebbing, cell shrinkage, nuclear fragmentation, chromatin condensation, chromosomal DNA fragmentation, and global mRNA decay. In some aspects, early stages of apoptosis can be indicated by activation of certain caspases, e.g., 2, 8, 9, and 10. In some aspects, middle to late stages of apoptosis are characterized by further loss of membrane integrity, chromatin condensation and DNA fragmentation, include biochemical events such as activation of caspases 3, 6, and 7.

[0175] In particular embodiments, the phenotype is negative expression of one or more factors associated with programmed cell death, for example pro-apoptotic factors known to initiate apoptosis, e.g., members of the death receptor pathway, activated members of the mitochondrial (intrinsic) pathway, such as Bcl-2 family members, e.g., Bax, Bad, and Bid, and caspases. In certain embodiments, the phenotype is the absence of an indicator, e.g., an Annexin V molecule or by TUNEL staining, that willpreferentially bind to cells undergoing apoptosis when incubated with or contacted to a cell composition. In some embodiments, the phenotype is or includes the expression of one or more markers that are indicative of an apoptotic state in the cell. In some embodiments, the phenotype is lack of expression and / or activation of a caspase, such as caspase 3. In some aspects, activation of caspase-3 is indicative of an increase or revival of apoptosis. In certain embodiments, caspase activation can be detected by known methods. In some embodiments, an antibody that binds specifically to an activated caspase (i.e., binds specifically to the cleaved polypeptide) can be used to detect caspase activation. In particular embodiments, the phenotype is or includes active caspase 3-. In some embodiments, the marker of apoptosis is a reagent that detects a feature in a cell that is associated with apoptosis. In certain embodiments, the reagent is an annexin V molecule.

[0176] In some embodiments, the compositions containing the engineered cells for administration contain a certain number or amount of cells that exhibit phenotypes indicative of or consistent with cell health. In some 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 the dose of engineered T cells express a marker of apoptosis, optionally Annexin V or active Caspase 3. In some embodiments, less than 5%, 4%, 3%, 2% or 1% of the CAR-expressing T cells in the dose of engineered T cells express Annexin V or active Caspase 3.

[0177] In some embodiments, the cells, binding molecules, or recombinant receptors are administered as part of a combination treatment, such as simultaneously with or sequentially with, in any order, another therapeutic intervention, such as another antibody or engineered cell or receptor or agent, such as a cytotoxic or therapeutic agent.

[0178] The cells, binding molecules and / or recombinant receptors in some embodiments are co- administered with one or more additional therapeutic agents or in connection with another therapeutic intervention, either simultaneously or sequentially in any order. In some contexts, the cells are co- administered with another therapy sufficiently close in time such that the cell populations enhance the effect of one or more additional therapeutic agents, or vice versa. In some embodiments, the cells, binding molecules and / or recombinant receptors are administered prior to the one or more additional therapeutic agents. In some embodiments, the cells, binding molecules and / or recombinant receptors are administered after to the one or more additional therapeutic agents. B. Response, Efficacy, and Survival

[0179] In some embodiments, the dose and / or frequency of administration is determined based on efficacy and / or response. In some embodiments, efficacy is determined by evaluating disease status. Exemplary methods for assessing disease status include: measurement of M protein in biological fluids, such as blood and / or urine, by electrophoresis and immunofixation; quantification of sFLC (κ and λ) in blood; skeletal survey; and imaging by positron emission tomography (PET) / computed tomography (CT)in subjects with extramedullary disease. In some embodiments, disease status can be evaluated by bone marrow examination. In some examples, dose and / or frequency of administration is determined by the expansion and persistence of the recombinant receptor or cell in the blood and / or bone marrow. In some embodiments, dose and / or frequency of administration is determined based on the antitumor activity of the recombinant receptor or engineered cell. In some embodiments antitumor activity is determined by the overall response rate (ORR) and / or International Myeloma Working Group (IMWG) Uniform Response Criteria (see Kumar et al. (2016) Lancet Oncol 17(8):e328-346). In some embodiments, response is evaluated 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 that have a MRD-negative disease include those exhibiting Absence of aberrant clonal plasma cells on bone marrow aspirate, ruled out by an assay with a minimum sensitivity of 1 in 105nucleated cells or higher (i.e., 10-5sensitivity), such as flow cytometry (next- generation flow cytometry; NGF) or high-throughput sequencing, e.g., deep sequencing or next- generation sequencing (NGS).

[0180] In some aspects, sustained MRD-negative includes subjects that exhibit MRD negativity in the marrow (NGF or NGS, or both) and by imaging as defined below, confirmed minimum of 1 year apart. Subsequent evaluations can be used to further specify the duration of negativity (e.g., MRD- negative at 5 years). In some aspects, flow MRD-negative includes subjects that exhibit an absence of phenotypically aberrant clonal plasma cells by NGF on bone marrow aspirates using the EuroFlow standard operation procedure for MRD detection in multiple myeloma (or validated equivalent method) with a minimum sensitivity of 1 in 105nucleated cells or higher. In some aspects, sequencing MRD- negative includes subjects that exhibit an absence of clonal plasma cells by NGS on bone marrow aspirate in which presence of a clone is defined as less than two identical sequencing reads obtained after DNA sequencing of bone marrow aspirates using the LymphoSIGHT platform (or validated equivalent method) with a minimum sensitivity of 1 in 105nucleated cells or higher. In some aspects, imaging plus MRD-negative includes subjects that exhibit MRD negativity as assessed by NGF or NGS plus disappearance of every area of increased tracer uptake found at baseline or a preceding PET / CT or decrease to less mediastinal blood pool SUV or decrease to less than that of surrounding normal tissue (see Kumar et al. (2016) Lancet Oncol 17(8):e328-346).

[0181] In some embodiments, response is evaluated based on the duration of response following administration of the recombinant receptor or cells. In some examples, dose and / or frequency of administration can be based on toxicity. In some embodiments, dose and / or frequency can be determined based on health-related quality of life (HRQoL) of the subject to which the recombinant receptor and / or cells is / are administered. In some embodiments, dose and / or frequency of administration can be changed, i.e., increased or decreased, based on any of the above criteria.

[0182] In some aspects, survival of the subject, survival within a certain time period, extent of survival, presence or duration of event-free or symptom-free survival, or relapse-free survival, is assessed. In some embodiments, any symptom of the disease or condition is assessed. In some embodiments, the measure of tumor burden is specified. In some embodiments, exemplary parameters for determination include particular clinical outcomes indicative of amelioration or improvement in the 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) Uniform 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. Specific thresholds for the parameters can be set to determine the efficacy of the methods provided herein. In some embodiments, the disease or disorder to be treated is multiple myeloma. In some embodiments, measurable disease criteria for multiple myeloma can include (1) serum M-protein 1 g / dL or greater; (2) Urine M-protein 200 mg or greater / 24 hour; (3) involved serum free light chain (sFLC) level 10 mg / dL or greater, with abnormal κ to λ ratio. In some cases, light chain disease is acceptable only for subjects without measurable disease in the serum or urine.

[0183] In some aspects, the response to the therapy, e.g., according to the provided embodiments, can be measured at a designated timepoint after the initiation of administration of the cell therapy. In some embodiments, the designated timepoint is at or about 1, 2, 3, 6, 9, 12, 18, 24, 30 or 36 months following initiation of the administration, or within a range defined by any of the foregoing. In some embodiments, the designated time point is 4, 8, 12, 16, 20, 24, 28, 32, 36, 48 or 52 weeks months following initiation of the administration, or within a range defined by any of the foregoing. In some embodiments, the designated timepoint is at or about 1 month following initiation of the administration. In some embodiments, the designated timepoint is at or about 3 months following initiation of the administration. In some embodiments, the designated timepoint is at or about 6 months following initiation of the administration. In some embodiments, the designated timepoint is at or about 9 months following initiation of the administration. In some embodiments, the designated timepoint is at or about 12 months following initiation of the administration.

[0184] In some embodiments, the response or outcome determined at or about 3, 6, 9 or 12 months after the designated timepoint is equal to or improved compared to the response or outcome determined at the initial designated timepoint. For example, in some aspects, if the response or outcome determined at the initial designated timepoint is stable disease (SD), Progressive disease (PD) or relapse, the subject treated according to the provided embodiments can show an equal or improved response or outcome (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) at a subsequent time point, after at or about 3, 6, 9 or 12 months after the initial designated timepoint, that is equal to the response or outcome at the initial designated timepoint, or a response or outcome that is objective response (OR), complete response (CR), stringent complete response (sCR), very good partial response (VGPR) or partial response (PR). In some aspects, subjects treated according to the provided embodiments can show a response or outcome that is improved between two time point of determination. In some aspects, the subject can exhibit a PR or VGPR in the initial designated timepoint for assessment, e.g., at 4 weeks after the initiation of administration, then exhibit an improved response, such as a CR or an sCR, at a later time point, e.g., at 12 weeks after the initiation of administration. In some respects, progression-free survival (PFS) is described as the length of time during and after the treatment of a disease, such as cancer, that a subject lives with the disease but it does not get worse. In some aspects, objective response (OR) is described as a measurable response. In some aspects, objective response rate (ORR; also known in some cases as overall response rate) is described as the proportion of patients who achieved CR or PR. In some aspects, overall survival (OS) is described as the length of time from either the date of diagnosis or the start of treatment for a disease, such as cancer, that subjects diagnosed with the disease are still alive. In some aspects, event-free survival (EFS) is described as the length of time after treatment for a cancer ends that the subject remains free of certain complications or events that the treatment was intended to prevent or delay. These events may include the return of the cancer or the onset of certain symptoms, such as bone pain from cancer that has spread to the bone, or death.

[0185] In some embodiments, the measure of duration of response (DOR) includes the time from documentation of tumor response to disease progression. In some embodiments, the parameter for assessing response can include durable response, e.g., response that persists after a period of time from initiation of therapy. In some embodiments, durable response is indicated by the response rate at 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 or outcome is durable for greater than at or about 3, 6, 9 or 12 months.

[0186] In some embodiments, the Eastern Cooperative Oncology Group (ECOG) performance status indicator can be used to assess or select subjects for treatment, e.g., subjects who have had poor performance from prior therapies (see, e.g., Oken et al. (1982) Am J Clin Oncol.5:649-655). The ECOG Scale of Performance Status describes a patient’s level of functioning in terms of their ability to care for themselves, daily activity, and physical ability (e.g., walking, working, etc.). In some embodiments, an ECOG performance status of 0 indicates that a subject can perform normal activity. In some aspects, subjects with an ECOG performance status of 1 exhibit some restriction in physical activity but the subject is fully ambulatory. In some aspects, patients with an ECOG performance status of 2 is more than 50% ambulatory. In some cases, the subject with an ECOG performance status of 2 may also be capable of self care; see e.g., Sørensen et al., (1993) Br J Cancer 67(4) 773-775. In some embodiments,the subject that are to be administered according to the methods or treatment regimen provided herein include those with an ECOG performance status of 0 or 1.

[0187] In some embodiments, the administration in accord with the provided methods effectively treats the subject despite the subject having become resistant to another therapy. In some embodiments, when administered to subjects according to the embodiments described herein, the dose or the composition is capable of achieving objective response (OR), in at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of subjects that were administered. In some embodiments, OR includes subjects who achieve stringent complete response (sCR), complete response (CR), very good partial response (VGPR), partial response (PR) and minimal response (MR). In some embodiments, when administered to subjects according to the embodiments described herein, the dose or the composition is capable of achieving stringent complete response (sCR), complete response (CR), very good partial response (VGPR) or partial response (PR), in at least 50%, 60%, 70%, 80%, or 85% of subjects that were administered. In some embodiments, when administered to subjects according to the embodiments described herein, the dose or the composition is capable of achieving stringent complete response (sCR) or complete response (CR) at least 20%, 30%, 40% 50%, 60% or 70% of subjects that were administered. In some aspects, particular response to the treatment, e.g., 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).

[0188] In some embodiments, the administration in accord with the provided methods effectively treats the subject despite the subject having become resistant to another therapy. In some embodiments, at least 30%, at least 35%, at least 40% at least 50%, at least 60%, at least 70%, or at least 80%, of subjects treated according to the method achieve complete remission (CR). In some embodiments, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least 80%, or at least 90% of the subjects treated according to the method achieve an objective response (OR). In some embodiments, at least or at least about 50% of subjects, at least or at least about 60% of the subjects, at least or at least about 70% of the subjects, at least or at least about 80% of the subjects or at least or at least about 90% of the subjects treated according to the method achieve CR and / or achieve an objective response (OR). In some embodiments, criteria assessed for effective treatment includes overall response rate (ORR; also known in some cases as objective response rate), complete response (CR; also known in some cases as complete remission), duration of response (DOR), progression-free survival (PFS), and / or overall survival (OS).

[0189] In some embodiments, at least 40% or at least 50% of subjects treated according to the methods provided herein achieve complete remission (CR; also known in some cases as complete response), exhibit progression-free survival (PFS) and / or overall survival (OS) of greater than at or about 3 months, 6 months or 12 months or greater than 13 months or approximately 14 months; on average,subjects treated according to the method exhibit a median PFS or OS of greater than at or about 6 months, 12 months, or 18 months; and / or the subject exhibits PFS or OS following therapy for at least at or about 6, 12, 18 or more months or longer.

[0190] In some embodiments, the subjects treated according to the provided methods exhibits a CRR of at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In some embodiments, the complete response rate (CRR) is calculated as the percentage of subjects with the best overall response (BOR) up to 12 months, up to 18 months, up to 24 months, up to 36 months or longer. C. Toxicity

[0191] In some embodiments, the provided methods are designed to or include features that result in a lower rate and / or lower degree of toxicity, toxic outcome or symptom, toxicity-promoting profile, factor, or property, such as a symptom or outcome associated with or indicative of cytokine release syndrome (CRS) or neurotoxicity (NT), for example, compared to administration of an alternative cell therapy, such as an alternative CAR+T cell composition and / or an alternative dosing of cells, e.g. a dosing of cells that is not administered at a defined ratio. Cytokine release syndrome (CRS) and neurotoxicity) can be graded according to the American Society for Transplantation and Cellular Therapy (ASTCT) Consensus Grading System (see e.g., Lee et al. Biol Blood Marrow Transplant.2019 Apr;25(4):625-38)).

[0192] In some aspects, although the lower differentiation state of the engineered T cells administered as part of the methods provided herein (e.g., the higher proportion of engineered T cells having a naïve-like or central memory phenotype, such as a phenotype selected from CCR7+CD45RA+, CD27+CCR7+, or CD62L-CCR7+) are expected to be more active than cells that are more differentiated, findings indicate that safety of the cell therapy can be successfully managed. In some aspects, providing a lower dose of the composition, e.g. compared to a cell composition produced by a process in which the cells are more differentiated, such as a process that includes expansion of the cells, achieves robust efficacy and high safety. In some aspects, it is found that even higher doses of cells of the provided anti- BCMA CAR compositions can be administered while maintaining alower degree of toxicity, such as a severe cytokine release syndrome (CRS) or severe neurotoxicity. Thus, the provided methods in some embodiments include the administration of higher doses of engineered T cells (e.g., greater than 50 × 106CAR-expressing T cells, such as at or about 100 × 106CAR-expressing T cells, 160 × 106CAR- expressing T cells, or 200 × 106CAR-expressing T cells), compared to methods that include the administration of an alternative cell therapy, such as an alternative CAR+T cell composition with engineered T cells that are more differentiated than those administered herein.

[0193] In some embodiments, the provided methods do not result in a high rate or likelihood of toxicity or toxic outcomes, or reduces the rate or likelihood of toxicity or toxic outcomes, such asneurotoxicity (NT), cytokine release syndrome (CRS), such as compared to certain other cell therapies. In some embodiments, the methods do not result in, or do not increase the risk of, severe NT (sNT), severe CRS (sCRS), macrophage activation syndrome, tumor lysis syndrome, fever of at least at or about 38 degrees Celsius for three or more days and a plasma level of CRP of at least at or about 20 mg / dL. In some embodiments, greater than or greater than about 30%, 35%, 40%, 50%, 55%, 60% or more of the subjects treated according to the provided methods do not exhibit any grade of CRS or any grade of neurotoxcity. In some embodiments, no more than 50% of subjects treated (e.g. at least 60%, at least 70%, at least 80%, at least 90% or more of the subjects treated) exhibit a cytokine release syndrome (CRS) higher than grade 2 and / or a neurotoxicity higher than grade 2. In some embodiments, at least 50% of subjects treated according to the method (e.g. at least 60%, at least 70%, at least 80%, at least 90% or more of the subjects treated) do not exhibit a severe toxic outcome (e.g. severe CRS or severe neurotoxicity), such as do not exhibit grade 3 or higher neurotoxicity and / or does not exhibit severe CRS, or does not do so within a certain period of time following the treatment, such as within a week, two weeks, or one month of the administration of the cells. In some embodiments, parameters assessed to determine certain toxicities include adverse events (AEs), dose-limiting toxicities (DLTs), CRS and NT.

[0194] Administration of adoptive T cell therapy, such as treatment with T cells expressing chimeric antigen receptors, can induce toxic effects or outcomes such as cytokine release syndrome and neurotoxicity. In some examples, such effects or outcomes parallel high levels of circulating cytokines, which may underlie the observed toxicity.

[0195] In some aspects, the toxic outcome is or is associated with or indicative of cytokine release syndrome (CRS) or severe CRS (sCRS). CRS, e.g., sCRS, can occur in some cases following adoptive T cell therapy and administration to subjects of other biological products. See Davila et al., Sci Transl Med 6, 224ra25 (2014); Brentjens et al., Sci. Transl. Med.5, 177ra38 (2013); Grupp et al., N. Engl. J. Med. 368, 1509–1518 (2013); and Kochenderfer et al., Blood 119, 2709–2720 (2012); Xu et al., Cancer Letters 343 (2014) 172-78.

[0196] Typically, CRS is caused by an exaggerated systemic immune response mediated by, for example, T cells, B cells, NK cells, monocytes, and / or macrophages. Such cells may release a large amount of inflammatory mediators such as cytokines and chemokines. Cytokines may trigger an acute inflammatory response and / or induce endothelial organ damage, which may result in microvascular leakage, heart failure, or death. Severe, life-threatening CRS can lead to pulmonary infiltration and lung injury, renal failure, or disseminated intravascular coagulation. Other severe, life-threatening toxicities can include cardiac toxicity, respiratory distress, neurologic toxicity and / or hepatic failure. In some aspects, fever, especially high fever (≥ 38.5°C or ≥ 101.3°F), is associated with CRS or risk thereof. In some cases, features or symptoms of CRS mimic infection. In some embodiments, infection is also considered in subjects presenting with CRS symptoms, and monitoring by cultures and empiric antibiotictherapy can be administered. Other symptoms associated with CRS can include cardiac dysfunction, adult respiratory distress syndrome, renal and / or hepatic failure, coagulopathies, disseminated intravascular coagulation, and capillary leak syndrome.

[0197] CRS may be treated using anti-inflammatory therapy such as an anti-IL-6 therapy, e.g., anti- IL-6 antibody, e.g., tocilizumab, or antibiotics or other agents as described. Outcomes, signs and symptoms of CRS are known and include those described herein. In some embodiments, where a particular dosage regimen or administration effects or does not effect a given CRS-associated outcome, sign, or symptom, particular outcomes, signs, and symptoms and / or quantities or degrees thereof may be specified.

[0198] In the context of administering CAR-expressing cells, CRS typically occurs 6-20 days after infusion of cells that express a CAR. See Xu et al., Cancer Letters 343 (2014) 172-78. In some cases, CRS occurs less than 6 days or more than 20 days after CAR T cell infusion. The incidence and timing of CRS may be related to baseline cytokine levels or tumor burden at the time of infusion. Commonly, CRS involves elevated serum levels of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, and / or interleukin (IL)-2. Other cytokines that may be rapidly induced in CRS are IL-1β, IL-6, IL-8, and IL-10.

[0199] Exemplary outcomes associated with CRS include fever, rigors, chills, hypotension, dyspnea, acute respiratory distress syndrome (ARDS), encephalopathy, ALT / AST elevation, renal failure, cardiac disorders, hypoxia, neurologic disturbances, and death. Neurological complications include delirium, seizure-like activity, confusion, word-finding difficulty, aphasia, and / or becoming obtunded. Other CRS- related outcomes include fatigue, nausea, headache, seizure, tachycardia, myalgias, rash, acute vascular leak syndrome, liver function impairment, and renal failure. In some aspects, CRS is associated with an increase in one or more factors such as serum-ferritin, d-dimer, aminotransferases, lactate dehydrogenase and triglycerides, or with hypofibrinogenemia or hepatosplenomegaly. Other exemplary signs or symptoms associated with CRS include hemodynamic instability, febrile neutropenia, increase in serum C-reactive protein (CRP), changes in coagulation parameters (for example, international normalized ratio (INR), prothrombin time (PTI) and / or fibrinogen), changes in cardiac and other organ function, and / or absolute neutrophil count (ANC).

[0200] In some embodiments, outcomes associated with CRS include one or more of: persistent fever, e.g., fever of a specified temperature, e.g., greater than at or about 38 degrees Celsius, for two or more, e.g., three or more, e.g., four or more days or for at least three consecutive days; fever greater than at or about 38 degrees Celsius; elevation of cytokines, such as a max fold change, e.g., of at least at or about 75, compared to pre-treatment levels of at least two cytokines (e.g., at least two of the group consisting of interferon gamma (IFNγ), GM-CSF, IL-6, IL-10, Flt-3L, fracktalkine, and IL-5, and / or tumor necrosis factor alpha (TNFα)), or a max fold change, e.g., of at least at or about 250 of at least one of such cytokines; and / or at least one clinical sign of toxicity, such as hypotension (e.g., as measured byat least one intravenous vasoactive pressor); hypoxia (e.g., plasma oxygen (PO2) levels of less than at or about 90%); and / or one or more neurologic disorders (including mental status changes, obtundation, and seizures). In some embodiments, neurotoxicity (NT) can be observed concurrently with CRS.

[0201] Exemplary CRS-related outcomes include increased or high serum levels of one or more factors, including cytokines and chemokines and other factors associated with CRS. Exemplary outcomes further include increases in synthesis or secretion of one or more of such factors. Such synthesis or secretion can be by the T cell or a cell that interacts with the T cell, such as an innate immune cell or B cell.

[0202] In some embodiments, the CRS-associated serum factors or CRS-related outcomes include inflammatory cytokines and / or chemokines, including interferon gamma (IFN-γ), TNF-a, IL-1β, IL-2, IL- 6, IL-7, IL-8, IL-10, IL-12, sIL-2Ra, granulocyte macrophage colony stimulating factor (GM-CSF), macrophage inflammatory protein (MIP)-1, tumor necrosis factor alpha (TNFα), IL-6, and IL-10, IL-1β, IL-8, IL-2, MIP-1, Flt-3L, fracktalkine, and / or IL-5. In some embodiments, the factor or outcome includes C reactive protein (CRP). In addition to being an early and easily measurable risk factor for CRS, CRP also is a marker for cell expansion. In some embodiments, subjects that are measured to have high levels of CRP, such as ≥ 15 mg / dL, have CRS. In some embodiments, subjects that are measured to have high levels of CRP do not have CRS. In some embodiments, a measure of CRS includes a measure of CRP and another factor indicative of CRS.

[0203] In some embodiments, one or more inflammatory cytokines or chemokines are monitored before, during, or after CAR treatment. In some aspects, the one or more cytokines or chemokines include IFN-γ, TNF-α, IL-2, IL-1β, IL-6, IL-7, IL-8, IL-10, IL-12, sIL-2Rα, granulocyte macrophage colony stimulating factor (GM-CSF), or macrophage inflammatory protein (MIP). In some embodiments, IFN-γ, TNF-α, and IL-6 are monitored.

[0204] CRS criteria that appear to correlate with the onset of CRS to predict which patients are more likely to be at risk for developing sCRS have been developed (see Davilla et al. Science translational medicine.2014;6(224):224ra25). Factors include fevers, hypoxia, hypotension, neurologic changes, elevated serum levels of inflammatory cytokines, such as a set of seven cytokines (IFNγ, IL-5, IL-6, IL- 10, Flt-3L, fractalkine, and GM-CSF) whose treatment-induced elevation can correlate well with both pretreatment tumor burden and sCRS symptoms. Other guidelines on the diagnosis and management of CRS are known (see e.g., Lee et al, Blood.2014;124(2):188-95; Lee et al., Biol Blood Marrow Transplant 2019; 25(4):625-38). In some embodiments, the criteria reflective of CRS grade are those detailed in Table 2 below.

[0205] In some embodiments, a criteria reflective of CRS grade are those detailed in Table 3 below.

[0206] In some embodiments, high-dose vasopressor therapy include those described in Table 4 below.

[0207] In some embodiments, the toxic outcome is a severe CRS. In some embodiments, the toxic outcome is the absence of severe CRS (e.g. moderate or mild CRS). In some embodiments, a subject is deemed to develop “severe CRS” (“sCRS”) in response to or secondary to administration of a cell therapy or dose of cells thereof, if, following administration, the subject displays: (1) fever of at least 38 degrees Celsius for at least three days; (2) cytokine elevation that includes either (a) a max fold change of at least 75 for at least two of the following group of seven cytokines compared to the level immediately following the administration: interferon gamma (IFNγ), GM-CSF, IL-6, IL-10, Flt-3L, fracktalkine, and IL-5 and / or (b) a max fold change of at least 250 for at least one of the following group of seven cytokines compared to the level immediately following the administration: interferon gamma (IFNγ), GM-CSF, IL-6, IL-10, Flt-3L, fracktalkine, and IL-5; and (c) at least one clinical sign of toxicity such as hypotension (requiring at least one intravenous vasoactive pressor) or hypoxia (PO2< 90%) or one or more neurologic disorder(s) (including mental status changes, obtundation, and / or seizures). In some embodiments, severe CRS includes CRS with a grade of 3 or greater, such as set forth in Table 2 and Table 3.

[0208] In some embodiments, the level of the toxic outcome, e.g. the CRS-related outcome, e.g. the serum level of an indicator of CRS, is measured by ELISA. In some embodiments, fever and / or levels of C-reactive protein (CRP) can be measured. In some embodiments, subjects with a fever and a CRP ≥ 15 mg / dL may be considered high-risk for developing severe CRS. In some embodiments, the CRS- associated serum factors or CRS-related outcomes include an increase in the level and / or concentration of inflammatory cytokines and / or chemokines, including Flt-3L, fracktalkine, granulocyte macrophage colony stimulating factor (GM-CSF), interleukin-1 beta (IL-1β), IL-2, IL-5, IL-6, IL-7, IL-8, IL-10, IL- 12, interferon gamma (IFN-γ), macrophage inflammatory protein (MIP)-1, MIP-1, sIL-2Rα, or tumor necrosis factor alpha (TNFα). In some embodiments, the factor or outcome includes C reactive protein (CRP). In addition to being an early and easily measurable risk factor for CRS, CRP also is a marker for cell expansion. In some embodiments, subjects that are measured to have high levels of CRP, such as ≥15 mg / dL, have CRS. In some embodiments, subjects that are measured to have high levels of CRP do not have CRS. In some embodiments, a measure of CRS includes a measure of CRP and another factor indicative of CRS.

[0209] In some embodiments, outcomes associated with severe CRS or grade 3 CRS or greater, such as grade 4 or greater, include one or more of: persistent fever, e.g., fever of a specified temperature, e.g., greater than at or about 38 degrees Celsius, for two or more, e.g., three or more, e.g., four or more days or for at least three consecutive days; fever greater than at or about 38 degrees Celsius; elevation of cytokines, such as a max fold change, e.g., of at least at or about 75, compared to pre-treatment levels of at least two cytokines (e.g., at least two of the group consisting of interferon gamma (IFNγ), GM-CSF, IL-6, IL-10, Flt-3L, fracktalkine, and IL-5, and / or tumor necrosis factor alpha (TNFα)), or a max fold change, e.g., of at least at or about 250 of at least one of such cytokines; and / or at least one clinical sign of toxicity, such as hypotension (e.g., as measured by at least one intravenous vasoactive pressor); hypoxia (e.g., plasma oxygen (PO2) levels of less than at or about 90%); and / or one or more neurologic disorders (including mental status changes, obtundation, and seizures). In some embodiments, severe CRS includes CRS that requires management or care in the intensive care unit (ICU).

[0210] In some embodiments, the CRS, such as severe CRS, encompasses a combination of (1) persistent fever (fever of at least 38 degrees Celsius for at least three days) and (2) a serum level of CRP of at least at or about 20 mg / dL. In some embodiments, the CRS encompasses hypotension requiring the use of two or more vasopressors or respiratory failure requiring mechanical ventilation. In some embodiments, the dosage of vasopressors is increased in a second or subsequent administration.

[0211] In some embodiments, severe CRS or grade 3 CRS encompasses an increase in alanine aminotransferase, an increase in aspartate aminotransferase, chills, febrile neutropenia, headache, left ventricular dysfunction, encephalopathy, hydrocephalus, and / or tremor.

[0212] The method of measuring or detecting the various outcomes may be specified.

[0213] In some aspects, the toxic outcome is or is associated with neurotoxicity. In some embodiments, symptoms associated with a clinical risk of neurotoxicity include confusion, delirium, aphasia, expressive aphasia, obtundation, myoclonus, lethargy, altered mental status, convulsions, seizure-like activity, seizures (optionally as confirmed by electroencephalogram (EEG)), elevated levels of beta amyloid (Aβ), elevated levels of glutamate, and elevated levels of oxygen radicals. In some embodiments, neurotoxicity is graded based on severity (e.g., using a Grade 1-5 scale (see, e.g., Guido Cavaletti & Paola Marmiroli Nature Reviews Neurology 6, 657-666 (December 2010); National Cancer Institute—Common Toxicity Criteria version 4.03 (NCI-CTCAE v4.03).

[0214] In some instances, neurologic symptoms may be the earliest symptoms of sCRS. In some embodiments, neurologic symptoms are seen to begin 5 to 7 days after cell therapy infusion. In some embodiments, duration of neurologic changes may range from 3 to 19 days. In some cases, recovery ofneurologic changes occurs after other symptoms of sCRS have resolved. In some embodiments, time or degree of resolution of neurologic changes is not hastened by treatment with anti-IL-6 and / or steroid(s).

[0215] In some embodiments, a subject is deemed to develop “severe neurotoxicity” in response to or secondary to administration of a cell therapy or dose of cells thereof, if, following administration, the subject displays symptoms that limit self-care (e.g. bathing, dressing and undressing, feeding, using the toilet, taking medications) from among: 1) symptoms of peripheral motor neuropathy, including inflammation or degeneration of the peripheral motor nerves; 2) symptoms of peripheral sensory neuropathy, including inflammation or degeneration of the peripheral sensory nerves, dysesthesia, such as distortion of sensory perception, resulting in an abnormal and unpleasant sensation, neuralgia, such as intense painful sensation along a nerve or a group of nerves, and / or paresthesia, such as functional disturbances of sensory neurons resulting in abnormal cutaneous sensations of tingling, numbness, pressure, cold and warmth in the absence of stimulus. In some embodiments, severe neurotoxicity includes neurotoxicity with a grade of 3 or greater, such as set forth in Table 5.

[0216] In some embodiments, the methods reduce symptoms associated with CRS or neurotoxicity compared to other methods. In some aspects, the provided methods reduce symptoms, outcomes or factors associated with CRS, including symptoms, outcomes or factors associated with severe CRS or grade 3 or higher CRS, compared to other methods. For example, subjects treated according to the present methods may lack detectable and / or have reduced symptoms, outcomes or factors of CRS, e.g. severe CRS or grade 3 or higher CRS, such as any described, e.g. set forth in Table 2 and Table 3. In some embodiments, subjects treated according to the present methods may have reduced symptoms of neurotoxicity, such as limb weakness or numbness, loss of memory, vision, and / or intellect, uncontrollable obsessive and / or compulsive behaviors, delusions, headache, cognitive and behavioral problems including loss of motor control, cognitive deterioration, and autonomic nervous systemdysfunction, and sexual dysfunction, compared to subjects treated by other methods. In some embodiments, subjects treated according to the present methods may have reduced symptoms associated with peripheral motor neuropathy, peripheral sensory neuropathy, dysethesia, neuralgia or paresthesia.

[0217] In some embodiments, the methods reduce outcomes associated with neurotoxicity including damages to the nervous system and / or brain, such as the death of neurons. In some aspects, the methods reduce the level of factors associated with neurotoxicity such as beta amyloid (Aβ), glutamate, and oxygen radicals.

[0218] In some embodiments, the toxicity outcome is a dose-limiting toxicity (DLT). In some embodiments, the toxic outcome is a dose-limiting toxicity. In some embodiments, the toxic outcome is the absence of a dose-limiting toxicity. In some embodiments, a dose-limiting toxicity (DLT) is defined as any grade 3 or higher toxicity as assessed by any known or published guidelines for assessing the particular toxicity, such as any described above and including the National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) version 4.0.

[0219] In some embodiments, the low rate, risk or likelihood of developing a toxicity, e.g. CRS or neurotoxicity or severe CRS or neurotoxicity, e.g. grade 3 or higher CRS or neurotoxicity, observed with administering a dose of T cells in accord with the provided methods, and / or with the provided articles of manufacture or compositions, permits administration of the cell therapy on an outpatient basis. In some embodiments, the administration of the cell therapy, e.g. dose of T cells (e.g. CAR+T cells) in accord with the provided methods, and / or with the provided articles of manufacture or compositions, is performed on an outpatient basis or does not require admission to the subject to the hospital, such as admission to the hospital requiring an overnight stay.

[0220] In some aspects, subjects administered the cell therapy, e.g. dose of T cells (e.g. CAR+T cells) in accord with the provided methods, and / or with the provided articles of manufacture or compositions, including subjects treated on an outpatient basis, are not administered an intervention for treating any toxicity prior to or with administration of the cell dose, unless or until the subject exhibits a sign or symptom of a toxicity, such as of a neurotoxicity or CRS. Exemplary agents for treating, delaying, attenuating or ameliorating a toxicity are described in Section II.

[0221] In some embodiments, if a subject administered the cell therapy, e.g. dose of T cells (e.g. CAR+T cells), including subjects treated on an outpatient basis, exhibits a fever the subject is given or is instructed to receive or administer a treatment to reduce the fever. In some embodiments, the fever in the subject is characterized as a body temperature of the subject that is (or is measured at) at or above a certain threshold temperature or level. In some aspects, the threshold temperature is that associated with at least a low-grade fever, with at least a moderate fever, and / or with at least a high-grade fever. In some embodiments, the threshold temperature is a particular temperature or range. For example, the threshold temperature may be at or about or at least at or about 38, 39, 40, 41, or 42 degrees Celsius, and / or may bea range of at or about 38 degrees Celsius to at or about 39 degrees Celsius, a range of at or about 39 degrees Celsius to at or about 40 degrees Celsius, a range of at or about 40 degrees Celsius to at or about 41 degrees, or a range of at or about 41 degrees Celsius to at or about 42 degrees Celsius.

[0222] In some embodiments, the treatment designed to reduce fever includes treatment with an antipyretic. An antipyretic may include any agent, e.g., compound, composition, or ingredient, that reduces fever, such as one of any number of agents known to have antipyretic effects, such as NSAIDs (such as ibuprofen, naproxen, ketoprofen, and nimesulide), salicylates, such as aspirin, choline salicylate, magnesium salicylate, and sodium salicylate, paracetamol, acetaminophen, Metamizole, Nabumetone, Phenaxone, antipyrine, febrifuges. In some embodiments, the antipyretic is acetaminophen. In some embodiments, acetaminophen can be administered at a dose of 12.5 mg / kg orally or intravenously up to every four hours. In some embodiments, it is or comprises ibuprofen or aspirin.

[0223] In some embodiments, if the fever is a sustained fever, the subject is administered an alternative treatment for treating the toxicity, such as any described in Section II below. For subjects treated on an outpatient basis, the subject is instructed to return to the hospital if the subject has and / or is determined to or to have a sustained fever. In some embodiments, the subject has, and / or is determined to or considered to have, a sustained fever if he or she exhibits a fever at or above the relevant threshold temperature, and where the fever or body temperature of the subject is not reduced, or is not reduced by or by more than a specified amount (e.g., by more than 1 °C, and generally does not fluctuate by about, or by more than about, 0.5 °C, 0.4 °C, 0.3 °C, or 0.2 °C), following a specified treatment, such as a treatment designed to reduce fever such as treatment with an antipyreticm, e.g. NSAID or salicylates, e.g. ibuprofen, acetaminophen or aspirin. For example, a subject is considered to have a sustained fever if he or she exhibits or is determined to exhibit a fever of at least at or about 38 or 39 degrees Celsius, which is not reduced by or is not reduced by more than at or about 0.5 °C, 0.4 °C, 0.3 °C, or 0.2 °C, or by at or about 1%, 2%, 3%, 4%, or 5%, over a period of 6 hours, over a period of 8 hours, or over a period of 12 hours, or over a period of 24 hours, even following treatment with the antipyretic such as acetaminophen. In some embodiments, the dosage of the antipyretic is a dosage ordinarily effective in such as subject to reduce fever or fever of a particular type such as fever associated with a bacterial or viral infection, e.g., a localized or systemic infection.

[0224] In some embodiments, the subject has, and / or is determined to or considered to have, a sustained fever if he or she exhibits a fever at or above the relevant threshold temperature, and where the fever or body temperature of the subject does not fluctuate by about, or by more than about, 1 °C, and generally does not fluctuate by about, or by more than about, 0.5 °C, 0.4 °C, 0.3 °C, or 0.2 °C. Such absence of fluctuation above or at a certain amount generally is measured over a given period of time (such as over a 24-hour, 12-hour, 8-hour, 6-hour, 3-hour, or 1-hour period of time, which may be measured from the first sign of fever or the first temperature above the indicated threshold). Forexample, in some embodiments, a subject is considered to or is determined to exhibit sustained fever if he or she exhibits a fever of at least at or about or at least at or about 38 or 39 degrees Celsius, which does not fluctuate in temperature by more than at or about 0.5°C, 0.4 °C, 0.3 °C, or 0.2 °C, over a period of 6 hours, over a period of 8 hours, or over a period of 12 hours, or over a period of 24 hours.

[0225] In some embodiments, the fever is a sustained fever; in some aspects, the subject is treated at a time at which a subject has been determined to have a sustained fever, such as within one, two, three, four, five six, or fewer hours of such determination or of the first such determination following the initial therapy having the potential to induce the toxicity, such as the cell therapy, such as dose of T cells, e.g. CAR+T cells.

[0226] In some embodiments, one or more interventions or agents for treating the toxicity, such as a toxicity-targeting therapies, is administered at a time at which or immediately after which the subject is determined to or confirmed to (such as is first determined or confirmed to) exhibit sustained fever, for example, as measured according to any of the aforementioned embodiments. In some embodiments, the one or more toxicity-targeting therapies is administered within a certain period of time of such confirmation or determination, such as within 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, or 8 hours thereof. II. CELL THERAPY AND ENGINEERING CELLS

[0227] In some embodiments, the cell therapy (e.g., T cell therapy) disclosed herein includes administering engineered cells expressing recombinant receptors (e.g. CAR) designed to recognize and / or specifically bind to antigens associated with the disease or condition, such as r / r / MM.

[0228] In some embodiments of the provided methods and uses, the engineered cells, such as T cells, express a chimeric receptors, such as a chimeric antigen receptors (CAR), that contains one or more domains that combine a ligand-binding domain (e.g. antibody or antibody fragment) that provides specificity for a desired antigen (e.g., tumor antigen) with intracellular signaling domains.

[0229] Among the provided embodiments are compositions, articles of manufacture, compounds, methods and uses including those targeting or directed to BCMA and BCMA-expressing cells and diseases. It is observed that BCMA is expressed, e.g., heterogeneously expressed, on certain diseases and conditions such as malignancies or tissues or cells thereof, e.g., on malignant plasma cells such as from all relapsed or newly diagnosed myeloma patients, for example, with little expression on normal tissues. Among the provided embodiments are approaches useful in the treatment of such diseases and conditions and / or for targeting such cell types, including nucleic acid molecules that encode BCMA-binding receptors, including chimeric antigen receptors (CARs), and the encoded receptors such as the encoded CARs, and compositions and articles of manufacture comprising the same. The receptors generally can contain antigen-binding domains that include antibodies (including antigen-binding antibody fragments,such as heavy chain variable (VH) regions, single domain antibody fragments and single chain fragments, including scFvs) specific for BCMA. Also provided are cells, such as engineered or recombinant cells expressing such BCMA-binding receptors, e.g., anti-BCMA CARs and / or containing nucleic acids encoding such receptors, and compositions and articles of manufacture and therapeutic doses containing such cells. Also provided are methods of evaluating, optimizing, making and using nucleic acid sequence(s), for example, nucleic acid sequences encoding recombinant BCMA-binding receptors. Also provided are methods of making and using (such as in the treatment or amelioration of BCMA- expressing diseases and conditions) cells (e.g., engineered cells) expressing or containing the recombinant BCMA-binding receptors and recombinant BCMA-binding receptor-encoding polynucleotides or compositions containing such cells.

[0230] Adoptive cell therapies (including those involving the administration of cells expressing chimeric receptors specific for a disease or disorder of interest, such as chimeric antigen receptors (CARs) and / or other recombinant antigen receptors, as well as other adoptive immune cell and adoptive T cell therapies) can be effective in the treatment of cancer and other diseases and disorders. In certain contexts, available approaches to adoptive cell therapy may not always be entirely satisfactory. In some aspects, the ability of the administered cells to recognize and bind to a target, e.g., target antigen such as BCMA, to traffic, localize to and successfully enter appropriate sites within the subject, tumors, and environments thereof, to become activated, expand, to exert various effector functions, including cytotoxic killing and secretion of various factors such as cytokines, to persist, including long-term, to differentiate, transition or engage in reprogramming into certain phenotypic states to provide effective and robust recall responses following clearance and re-exposure to target ligand or antigen, and avoid or reduce exhaustion, anergy, terminal differentiation, and / or differentiation into a suppressive state.

[0231] In some aspects, available approaches for treatment of diseases or disorders such as multiple myeloma is complex and may not always be entirely satisfactory. In some aspects, choosing a treatment regimen can depend on numerous factors including drug availability, response to prior therapy, aggressiveness of the relapse, eligibility for autologous stem cell transplantation (ASCT), and whether the relapse occurred on or off therapy. In some aspects, MM results in relapses and remissions, and existing regimen in some cases can result in relapse and / or toxicity from the treatment. In some cases, subjects with particularly aggressive disease, such as subjects that have persistent or relapsed disease after various therapies, subjects with a high disease burden, such as a high tumor burden, and / or subjects with particularly aggressive types of disease, such as plasmacytoma, can be particularly difficult to treat, and responses to certain therapies in these subjects can be poor or have a short duration. In some cases, subjects who have been heavily pre-treated, e.g., subjects who have relapsed after several different prior therapies, can exhibit a low response rate and / or high incidence of adverse events. In some aspects, the provided embodiments are based on an observation that treatment according to the providedembodiments results in a high response rate, low incidences of adverse events (e.g., toxicity), prolonged response, and in some cases, improvement in the response over time.

[0232] The provided embodiments, in some contexts, are based on an observation from a clinical study, that administration of engineered cells expressing a particular recombinant receptor, such as those described herein, results in a high response rate and a low rate of adverse events such as cytokine release syndrome (CRS) or neurological events (NE; or neurotoxicity; NT). In some aspects, the provided cells, methods and uses result in a cell therapy that exhibits prolonged persistence of the cells after administration of the cells, along with a high response rate and a low rate of toxicity (e.g., CRS or NE, such as grade 3 or higher CRS or grade 3 or higher neurotoxicity). In some aspects, such high response and low rate of toxicity (e.g., grade 3 or higher CRS or grade 3 or higher neurotoxicity), is achieved from employing various different doses of cells. For example, even at a relatively low dose of cells, a high rate of objective response and high level of response (e.g., very good partial response, VGPR, or better) is achieved. In some cases, a relatively high dose of cells can be administered, and such doses are observed to result in a high rate of objective response with low rate of toxicity (e.g., grade 3 or higher CRS or grade 3 or higher neurotoxicity). In some cases, the provided embodiments also permit improved expansion and / or persistence of the administered engineered cells, and in some cases result in prolonged response and / or response that is improved over time. In some aspects, treatment of subjects with aggressive or refractory disease (e.g., heavily pre-treated subjects, subjects with a high tumor burden and / or subjects with aggressive disease types) according to the provided embodiments, was observed to provide a safe, effective and durable treatment.

[0233] In some contexts, optimal response to therapy can depend on the ability of the engineered recombinant receptors such as CARs, to be consistently and reliably expressed on the surface of the cells and / or bind the target antigen. For example, in some cases, heterogeneity of the transcribed RNA from an introduced transgene (e.g., encoding the recombinant receptor) can affect the expression and / or activity of the recombinant receptor, in some cases when expressed in a cell, such as a human T cell, used in cell therapy. In some contexts, the length and type of spacer in the recombinant receptor, such as a CAR, can affect the expression, activity and / or function of the receptor.

[0234] Also, in some contexts, certain recombinant receptors can exhibit antigen-independent activity or signaling (also known as “tonic signaling”), which could lead to undesirable effects, such as due to increased differentiation and / or exhaustion of T cells that express the recombinant receptor. In some aspects, such activities may limit the T cell’s activity, effect or potency. In some cases, during engineering and ex vivo expansion of the cells for recombinant receptor expression, the cells may exhibit phenotypes indicative of exhaustion, due to tonic signaling through the recombinant receptor.

[0235] In some contexts, properties of particular target antigens that the recombinant receptors specifically bind, recognize or target, can that affect the activity of the receptor. In some contexts, B-cellmaturation antigen (BCMA), is typically expressed on malignant plasma cells and is an attractive therapeutic target for cell therapy. In some cases, BCMA is can be cleaved by gamma secretase, generating a soluble BCMA (sBCMA), or “shed” form of BCMA, reducing the BCMA expressed on the surface of target cells. In some cases, the activity of the BCMA-binding molecules, such as anti-BCMA chimeric antigen receptors, can be blocked or inhibited by the presence of soluble BCMA. Improved strategies are needed for optimal responses to cell therapies, in particular, for recombinant receptors that specifically bind, recognize or target BCMA, such as BCMA expressed on the surface of the target cells.

[0236] The provided embodiments, in some contexts, are based on the observation that particular spacers and optimization of the nucleic acid sequences can lead to consistent and robust expression of the recombinant receptor. The provided BCMA-binding recombinant receptors offer advantages over available approaches for cell therapies, in particular, BCMA-targeting cell therapy. In some embodiments, provided BCMA-binding recombinant receptors contain fully human antigen-binding domains, with low affinity for binding soluble BCMA. In some embodiments, provided BCMA-binding recombinant receptors contain a modified spacer that result in enhanced binding to BCMA expressed on the surface of target cells. In some embodiments, provided BCMA-binding recombinant receptors are observed to exhibit reduced antigen-independent, tonic signaling, which in some cases can result in reduced exhaustion of the cells from antigen-independent signaling, and lack of inhibition by soluble BCMA. In some embodiments, provided BCMA-binding recombinant receptors exhibit activity or potency against target cells that express a low density or low level of BCMA.

[0237] In various aspects, the provided BCMA-binding recombinant receptors, polynucleotides encoding such receptors, engineered cells and cell compositions, exhibit certain desired properties that can overcome or counteract certain limitations that can reduce optimal responses to cell therapy, for example, cell therapy with engineered cells expressing a BCMA-binding recombinant receptor. In some aspects, compositions containing engineered cells expressing an exemplary BCMA-binding recombinant receptor provided herein was observed to exhibit consistency of cell health of the engineered cells, and was associated with improved clinical response. In some aspects, compositions containing the engineered cells expressing an exemplary BCMA-binding recombinant receptor provided herein was observed to be enriched for immune cell subtypes, e.g., CD4+ or CD8+ T cell subtypes, that were associated with central memory T cell (TCM) phenotype, which, in some aspects is associated with increased persistence and durability of the engineered cells. In some contexts, the provided embodiments, including the recombinant receptors, polynucleotides encoding such receptors, engineered cells and cell compositions, can provide various advantages over available therapies targeting BCMA, to improve the activity of the recombinant receptors and response to BCMA-targeting cell therapies. In addition, the provided methods and uses of the engineered cells or compositions comprising the engineered cells, has been observed to provide an advantage in treating subjects, that results in a highresponse rate, a durable response, and low rate of adverse events, at various different dose levels tested. Further, the provided methods and uses of the engineered cells or compositions comprising the engineered cells, has been observed to provide an advantage in treating subjects with particularly aggressive and / or refractory disease, or subjects who have relapsed and / or are refractory to numerous different prior treatments for the disease. A. Chimeric Antigen Receptors

[0238] Provided in some aspects are BCMA-binding agents, such as cell surface proteins, such as recombinant receptors or chimeric antigen receptors that bind or recognize BCMA molecules and polynucleotides encoding BCMA-binding cell surface proteins, such as recombinant receptors (e.g., chimeric antigen receptors; CARs), and cells expressing such receptors. The BCMA-binding cell surface proteins generally contain antibodies (e.g., antigen-binding antibody fragments), and / or other binding peptides that specifically recognize, such as specifically bind to BCMA, such as to BCMA proteins, such as human BCMA protein. In some aspects, the agents bind to an extracellular portion of BCMA. Also provided are cells, e.g., engineered cells, comprising such polynucleotides or expressing such receptors, and compositions comprising such engineered cells. In some aspects, also provided are methods employing such cells and compositions, and uses thereof, such as in therapeutic methods.

[0239] In some embodiments, the polynucleotides are optimized, or contain certain features designed for optimization, such as for codon usage, to reduce RNA heterogeneity and / or to modify, e.g., increase or render more consistent among cell product lots, expression, such as surface expression, of the encoded receptor. In some embodiments, polynucleotides, encoding BCMA-binding cell surface proteins, are modified as compared to a reference polynucleotide, such as to remove cryptic or hidden splice sites, to reduce RNA heterogeneity. In some embodiments, polynucleotides, encoding BCMA-binding cell surface proteins, are codon optimized, such as for expression in a mammalian, e.g., human, cell such as in a human T cell. In some aspects, the modified polynucleotides result in in improved, e.g., increased or more uniform or more consistent level of, expression, e.g., surface expression, when expressed in a cell. Such polynucleotides can be utilized in constructs for generation of engineered cells that express the encoded BCMA-binding cell surface protein. Thus, also provided are cells expressing the recombinant receptors encoded by the polynucleotides provided herein and uses thereof in adoptive cell therapy, such as treatment of diseases and disorders associated with BCMA expression, such as multiple myeloma.

[0240] Among the provided polynucleotides are those that encode recombinant receptors, such as antigen receptors, that specifically recognize, such as specifically bind, BCMA, such as a human BCMA. In some aspects, the encoded receptors, such as those containing BCMA-binding polypeptides, and compositions and articles of manufacture and uses of the same, also are provided.

[0241] Among the BCMA-binding polypeptides are antibodies, such as single-chain antibodies (e.g., antigen binding antibody fragments), or portions thereof. In some examples, the recombinant receptors are chimeric antigen receptors, such as those containing anti-BCMA antibodies or antigen- binding fragments thereof. In any of the embodiments, an antibody or antigen binding fragment, in the provided CARs, that specifically recognizes an antigen, e.g. BCMA, specifically binds to the antigen. The provided polynucleotides can be incorporated into constructs, such as deoxyribonucleic acid (DNA) or RNA constructs, such as those that can be introduced into cells for expression of the encoded recombinant BCMA-binding receptors.

[0242] In some cases, the polynucleotide encoding the BCMA-binding receptor contains a signal sequence that encodes a signal peptide, in some cases encoded upstream of the nucleic acid sequences encoding the BCMA-binding receptor, or joined at the 5’ terminus of the nucleic acid sequences encoding the antigen-binding domain. In some cases, the polynucleotide containing nucleic acid sequences encoding the BCMA-binding receptor, e.g., chimeric antigen receptor (CAR), contains a signal sequence that encodes a signal peptide. In some aspects, the signal sequence may encode a signal peptide derived from a native polypeptide. In other aspects, the signal sequence may encode a heterologous or non-native signal peptide. In some cases, the polynucleotide encoding the BCMA- binding receptor can contain nucleic acid sequence encoding additional molecules, such as a surrogate marker or other markers, or can contain additional components, such as promoters, regulatory elements and / or multicistronic elements. In some embodiments, the nucleic acid sequence encoding the BCMA- binding receptor can be operably linked to any of the additional components.

[0243] The provided BCMA-binding receptors, e.g., expressed in the cells employed in the methods and uses provided herein, generally contain an extracellular binding molecule and an intracellular signaling domain. Among the provided binding molecules are polypeptides containing antibodies, including single chain cell surface proteins, e.g., recombinant receptors such as chimeric antigen receptors, containing such antibodies.

[0244] Among the provided binding molecules (e.g., BCMA-binding molecules) are single chain cell surface proteins, such as recombinant receptors (e.g., antigen receptors), that include one of the provided antibodies or fragment thereof (e.g., BCMA-binding fragment). The recombinant receptors include antigen receptors that specifically bind to or specifically recognize BCMA, such as antigen receptors containing the provided anti-BCMA antibodies, e.g., antigen-binding fragments. Among the antigen receptors are functional non-TCR antigen receptors, such as chimeric antigen receptors (CARs). Also provided are cells expressing the recombinant receptors and uses thereof in adoptive cell therapy, such as treatment of diseases and disorders associated with BCMA expression.

[0245] Exemplary antigen receptors, including CARs, and methods for engineering and introducing such antigen receptors into cells, include those described, for example, in international patent applicationpublication Nos. WO200014257, WO2013126726, WO2012 / 129514, WO2014031687, WO2013166321, WO2013071154, WO2013123061 U.S. patent application publication Nos. US2002131960, US2013287748, US20130149337, U.S. Patent Nos.6,451,995, 7,446,190, 8,252,592, 8,339,645, 8,398,282, 7,446,179, 6,410,319, 7,070,995, 7,265,209, 7,354,762, 7,446,191, 8,324,353, and 8,479,118, and European patent application No. EP2537416, and / or those described by Sadelain et al., Cancer Discov.2013 April; 3(4): 388–398; Davila et al. (2013) PLoS ONE 8(4): e61338; Turtle et al., Curr. Opin. Immunol., 2012 October; 24(5): 633-39; Wu et al., Cancer, 2012 March 18(2): 160-75. In some aspects, the antigen receptors include a CAR as described in U.S. Patent No.7,446,190, and those described in International Patent Application Publication No. WO2014055668. Exemplary CARs include CARs as disclosed in any of the aforementioned publications, such as WO2014031687, US 8,339,645, US 7,446,179, US 2013 / 0149337, US 7,446,190, and US 8,389,282, and in which the antigen- binding portion, e.g., scFv, is replaced by an antibody or an antigen-binding fragment thereof, as provided herein.

[0246] In some embodiments, the provided CAR has an amino acid sequence selected from among SEQ ID NOs: 15-20, or an amino acid sequence that exhibits at least or about at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence set forth in any of SEQ ID NOs 15-20. In some embodiments, the provided CAR has an amino acid sequence set forth in SEQ ID NO: 19, or an amino acid sequence that exhibits at least or about at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:19.

[0247] In some embodiments, the provided CAR is encoded by a polynucleotide, such as an polynucleotide with the nucleic acid sequence set forth in any of SEQ ID NOs 9-14, or a sequences that exhibits at least or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the nucleic acid sequence set forth in any of SEQ ID NOs: 9-14. In some embodiments, the provided CAR is encoded by a polynucleotide, such as an polynucleotide with the nucleic acid sequence set forth in any of SEQ ID NOs:13 and 14, or a sequences that exhibits at least or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the nucleic acid sequence set forth in any of SEQ ID NOs: 13 and 14. In some embodiments, the provided CAR is encoded by a polynucleotide, such as an polynucleotide with the nucleic acid sequence set forth in SEQ ID NO:13 or a sequences that exhibits at least or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto. In some embodiments, the provided CAR is encoded by a polynucleotide, such as an polynucleotide with the nucleic acid sequence set forth in SEQ ID NO:13.

[0248] In some embodiments, the nucleic acid encoding the antigen-binding domain comprises (a) the sequence of nucleotides set forth in any of SEQ ID NOS: 30, 31, 50, 51, 59, 60, 82, 84, 113, 115; (b) a sequence of nucleotides that has at least 90% sequence identity to any of SEQ ID NOS: 30, 31, 50, 51,59, 60, 82, 84, 113, 115; or (c) a degenerate sequence of (a) or (b). In some embodiments, the nucleic acid encoding the antigen-binding domain comprises (a) a sequence of nucleotides encoding the amino acid sequence set forth in any of SEQ ID NOS: 29, 49, 58, 83, 114, 127, 128, 129, 130; (b) a sequence of nucleotides that has at least 90% sequence identity to a sequence of nucleotides encoding the amino acid sequence set forth in any of SEQ ID NOS: 29, 49, 58, 83, 114, 126, 127, 128, 129, 130; or (c) a degenerate sequence of (a) or (b). 1. Antigen-binding domain

[0249] Among the chimeric receptors are chimeric antigen receptors (CARs). The chimeric receptors, such as CARs, generally include an extracellular antigen binding domain that includes, is, or is comprised within or comprises, one of the provided anti-BCMA antibodies. Thus, the chimeric receptors, e.g., CARs, typically include in their extracellular portions one or more BCMA-binding molecules, such as one or more antigen-binding fragment, domain, or portion, or one or more antibody variable regions, and / or antibody molecules, such as those described herein.

[0250] The term “antibody” herein is used in the broadest sense and includes polyclonal and monoclonal antibodies, including intact antibodies and functional (antigen-binding) antibody fragments, including fragment antigen binding (Fab) fragments, F(ab’)2 fragments, Fab’ fragments, Fv fragments, recombinant IgG (rIgG) fragments, heavy chain variable (VH) regions capable of specifically binding the antigen, single chain antibody fragments, including single chain variable fragments (scFv), and single domain antibodies (e.g., sdAb, sdFv, nanobody) 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 full-length antibodies, including antibodies of any class or sub-class, including IgG and sub-classes thereof, IgM, IgE, IgA, and IgD.

[0251] 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 within antibody variable regions, which confer antigen specificity and / or binding affinity. In general, there are three CDRs in each heavy chain variable region (CDR-H1, CDR-H2, CDR-H3) and three CDRs in each light chain variable region (CDR-L1, CDR-L2, CDR-L3). “Framework regions” and “FR” are known in the art to refer to the non-CDR portions of the variable regions of the heavy and light chains. In general, there are four FRs in each full-length heavy chain variable region (FR-H1, FR-H2, FR-H3, and FR-H4), and four FRs in each full-length light chain variable region (FR-L1, FR-L2, FR-L3, and FR-L4).

[0252] The precise amino acid sequence boundaries of a given CDR or FR can be readily determined using any of a number of well-known schemes, including those described by Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (“Kabat” numbering scheme); 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 (“IMGT” numbering scheme); Honegger A and Plückthun 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, (“Aho” numbering scheme); and Martin et al., “Modeling antibody hypervariable loops: a combined algorithm,” PNAS, 1989, 86(23):9268-9272, (“AbM” numbering scheme).

[0253] 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 alignments, while the Chothia scheme is based on structural information. Numbering for both the Kabat and Chothia schemes is based upon the most common antibody region sequence lengths, with insertions accommodated by insertion letters, for example, “30a,” and deletions appearing in some antibodies. The two schemes place certain insertions and deletions (“indels”) at different positions, resulting in differential numbering. The Contact scheme is based on analysis of complex crystal structures and is similar in many respects to the Chothia numbering scheme. The AbM scheme is a compromise between Kabat and Chothia definitions based on that used by Oxford Molecular’s AbM antibody modeling software.

[0254] Table 6, below, lists exemplary position boundaries of CDR-L1, CDR-L2, CDR-L3 and CDR-H1, CDR-H2, CDR-H3 as identified by Kabat, Chothia, AbM, and Contact schemes, respectively. For CDR-H1, residue numbering is listed using both the Kabat and Chothia numbering schemes. FRs are located between CDRs, for example, with FR-L1 located before CDR-L1, FR-L2 located between CDR- L1 and CDR-L2, FR-L3 located between CDR-L2 and CDR-L3 and so forth. It is noted that because the shown Kabat numbering scheme places insertions at H35A and H35B, the end of the Chothia CDR-H1 loop when numbered using the shown Kabat numbering convention varies between H32 and H34, depending on the length of the loop.1 - Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD 2 - Al-Lazikani et al., (1997) JMB 273,927-948

[0255] Thus, unless otherwise specified, a “CDR” or “complementary determining region,” or individual specified CDRs (e.g., CDR-H1, CDR-H2, CDR-H3), of a given antibody or region thereof, such as a variable region thereof, should be understood to encompass a (or the specific) complementary determining region as defined by any of the aforementioned schemes, or other known schemes. For example, where it is stated that a particular CDR (e.g., a CDR-H3) contains the amino acid sequence of a corresponding CDR in a given VH or VL region amino acid sequence, it is understood that such a CDR has a sequence of the corresponding CDR (e.g., CDR-H3) within the variable region, as defined by any of the aforementioned schemes, or other known schemes. In some embodiments, specific CDR sequences are specified. Exemplary CDR sequences of provided antibodies are described using various numbering schemes, although it is understood that a provided antibody can include CDRs as described according to any of the other aforementioned numbering schemes or other numbering schemes known to a skilled artisan.

[0256] Likewise, unless otherwise specified, a FR or individual specified FR(s) (e.g., FR-H1, FR- H2, FR-H3, FR-H4), of a given antibody or region thereof, such as a variable region thereof, should be understood to encompass a (or the specific) framework region as defined by any of the known schemes. In some instances, the scheme for identification of a particular CDR, FR, or FRs or CDRs is specified, such as the CDR as defined by the Kabat, Chothia, AbM, IMGT or Contact method, or other known schemes. In other cases, the particular amino acid sequence of a CDR or FR is given.

[0257] 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 antigen. The variable regions of the heavy chain and light chain (VHand VL, respectively) of a native antibody generally have similar structures, with each domain comprising four conserved framework regions (FRs) and three CDRs. (See, e.g., Kindt et al. Kuby Immunology, 6th ed., W.H. Freeman and Co., page 91 (2007). A single VHor VLdomain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind a particular antigen may be isolated using a VHor VLdomain from an antibody that binds the antigen to screen a library of complementary VL or VH domains, respectively. See, e.g., Portolano et al., J. Immunol.150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0258] Among the antibodies included in the provided CARs are antibody fragments. An “antibody fragment” or “antigen-binding fragment” refers to a molecule other than an intact antibody thatcomprises a portion of an intact antibody that binds the antigen to which the intact antibody binds. Examples of antibody fragments include but are not limited to Fv, Fab, Fab’, Fab’-SH, F(ab’)2; diabodies; linear antibodies; heavy chain variable (VH) regions, single-chain antibody molecules such as scFvs and single-domain antibodies comprising only the VHregion; and multispecific antibodies formed from antibody fragments. In some embodiments, the antigen-binding domain in the provided CARs is or comprises an antibody fragment comprising a variable heavy chain (VH) and a variable light chain (VL) region. In particular embodiments, the antibodies are single-chain antibody fragments comprising a heavy chain variable (VH) region and / or a light chain variable (VL) region, such as scFvs.

[0259] Single-domain antibodies (sdAbs) are antibody fragments comprising all or a portion of the heavy chain variable region or all or a portion of the light chain variable region of an antibody. In certain embodiments, a single-domain antibody is a human single-domain antibody.

[0260] Antibody fragments can be made by various techniques, including but not limited to proteolytic digestion of an intact antibody as well as production by recombinant host cells. In some embodiments, the antibodies are recombinantly-produced fragments, such as fragments comprising arrangements that do not occur naturally, such as those with two or more antibody regions or chains joined by synthetic linkers, e.g., peptide linkers, and / or that are may not be produced by enzyme digestion of a naturally-occurring intact antibody. In some aspects, the antibody fragments are scFvs.

[0261] 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 optionally may include at least a portion of an antibody constant region derived from a human antibody. A “humanized form” of a non-human antibody, refers to a variant of the non-human antibody that has undergone humanization, typically to reduce immunogenicity to humans, while retaining the specificity and affinity of the parental 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), e.g., to restore or improve antibody specificity or affinity.

[0262] Among the anti-BCMA antibodies included in the provided CARs are human antibodies. A “human antibody” is an antibody with an amino acid sequence corresponding to that of an antibody produced by a human or a human cell, or non-human source that utilizes human antibody repertoires or other human antibody-encoding sequences, including human antibody libraries. The term excludes humanized forms of non-human antibodies comprising non-human antigen-binding regions, such as those in which all or substantially all CDRs are non-human. The term includes antigen-binding fragments of human antibodies.

[0263] Human antibodies may be prepared by administering an immunogen to a transgenic animal that has been modified to produce intact human antibodies or intact antibodies with human variableregions in response to antigenic challenge. Such animals typically contain all or a portion of the human immunoglobulin loci, which replace the endogenous immunoglobulin loci, or which are present extrachromosomally or integrated randomly into the animal’s chromosomes. In such transgenic animals, the endogenous immunoglobulin loci have generally been inactivated. Human antibodies also may be derived from human antibody libraries, including phage display and cell-free libraries, containing antibody-encoding sequences derived from a human repertoire.

[0264] Among the antibodies included in the provided CARs are those that are monoclonal antibodies, including monoclonal antibody fragments. The term “monoclonal antibody” as used herein refers to an antibody obtained from or within a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical, except for possible variants containing naturally occurring mutations or arising during production of a monoclonal antibody preparation, such variants generally being present in minor amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different epitopes, each monoclonal antibody of a monoclonal antibody preparation is directed against a single epitope on an antigen. The term is not to be construed as requiring production of the antibody by any particular method. A monoclonal antibody may be made by a variety of techniques, including but not limited to generation from a hybridoma, recombinant DNA methods, phage-display and other antibody display methods.

[0265] In some embodiments, the CAR includes a BCMA-binding portion or portions of the antibody molecule, such as a heavy chain variable (VH) region and / or light chain variable (VL) region of the 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 an 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 any anti-BCMA antibody described or derived from any anti-BCMA antibody described. See, e.g., Carpenter et al., Clin Cancer Res., 2013, 19(8):2048-2060, WO 2016090320, WO2016090327, WO2010104949 and WO2017173256. 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 an antigen-binding domain that is an scFv containing a variable heavy (VH) and / or a variable light (VL) region derived from an antibody described in WO 2016090320 or WO2016090327.

[0266] In some embodiments, the antibody, e.g., the anti-BCMA antibody or antigen-binding fragment, contains a heavy and / or light chain variable (VHor VL) region sequence as described, or a sufficient antigen-binding portion thereof. In some embodiments, the anti-BCMA antibody, e.g., antigen-binding fragment, contains a VHregion sequence or sufficient antigen-binding portion thereof that contains a CDR-H1, CDR-H2 and / or CDR-H3 as described. In some embodiments, the anti-BCMA antibody, e.g., antigen-binding fragment, contains a VLregion sequence or sufficient antigen-binding portion that contains a CDR-L1, CDR-L2 and / or CDR-L3 as described. In some embodiments, the anti-BCMA antibody, e.g., antigen-binding fragment, contains a VHregion sequence that contains a CDR-H1, CDR-H2 and / or CDR-H3 as described and contains a VLregion sequence that contains a CDR-L1, CDR- L2 and / or CDR-L3 as described. Also among the antibodies are those having sequences at least at 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.

[0267] In some embodiments, the antibody is a single domain antibody (sdAb) comprising only a VH region sequence or a sufficient antigen-binding portion thereof, such as any of the above described VH sequences (e.g., a CDR-H1, a CDR-H2, a CDR-H3 and / or a CDR-H4).

[0268] In some embodiments, an antibody provided herein (e.g., an anti-BCMA antibody) or antigen-binding fragment thereof comprising a VH region further comprises a light chain or a sufficient antigen binding portion thereof. For example, in some embodiments, the antibody or antigen-binding fragment thereof contains a VH region and a VL region, or a sufficient antigen-binding portion of a VH and VL region. In such embodiments, a VH region sequence can be any of the above described VH sequence. In some such embodiments, the antibody is an antigen-binding fragment, such as a Fab or an scFv. In some such embodiments, the antibody is a full-length antibody that also contains a constant region.

[0269] In some embodiments, the antibody, e.g., antigen-binding fragment thereof, in the provided CAR, has a heavy chain variable (VH) region having the amino acid sequence selected from any one of SEQ ID NOs: 32, 52, 61, 85, 116, 125, 131, or an amino acid sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the VHregion amino acid selected from any one of SEQ ID NOs: 32, 52, 61, 85, 116, 125, 131, or contains a CDR-H1, CDR-H2, and / or CDR-H3 present in such a VHsequence. In some embodiments, the antibody or antibody fragment, in the provided CAR, has a VHregion of any of the antibodies or antibody binding fragments described in WO 2016 / 090327, WO 2016 / 090320, WO 2017 / 173256, or WO 2019 / 090003.

[0270] In some embodiments, the antibody, e.g., antigen-binding fragment thereof, in the provided CAR, has a light chain variable (VL) region having the amino acid sequence selected from any one of SEQ ID NOs: 33, 53, 62, 88, 119, 127, 132, or an amino acid sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the VLregion amino acid selected from any one of SEQ ID NOs: 33, 53, 62, 88, 119, 127, 132, or contains a CDR-L1, CDR-L2, and / or CDR-L3 present in such a VLsequence. In some embodiments, the antibody or antibody fragment, in the provided CAR, has a VLregion of any of the antibodies or antibody binding fragments described in WO 2016 / 090327, WO 2016 / 090320, WO 2017 / 173256, or WO 2019 / 090003.

[0271] In some embodiments, the antibody or antibody fragment, in the provided CAR, has a VH region and a VL region of any of the antibodies or antibody binding fragments described in WO 2016 / 090327, WO 2016 / 090320, WO 2017 / 173256, or WO 2019 / 090003. In some embodiments, theantibody or antibody fragment, in the provided CAR, is an scFv of any of the antibodies or antibody binding fragments described in WO 2016 / 090327, WO 2016 / 090320, WO 2017 / 173256, or WO 2019 / 090003.

[0272] In some embodiments, the BCMA-directed CAR is an anti-BCMA CAR as described in any one of WO 2016 / 090320, WO 2017 / 173256, or WO 2019 / 090003.

[0273] In some embodiments, the VHand VLregions of the antibody, e.g., antigen-binding fragment thereof, in the provided CAR, comprises: the amino acid sequence of SEQ ID NOS:32 and 33, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:32 and 33, respectively; the amino acid sequence of SEQ ID NOS:52 and 53, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:52 and 53, respectively; the amino acid sequence of SEQ ID NOS:61 and 62, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:61 and 62, respectively; the amino acid sequence of SEQ ID NOS:85 and 88, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:85 and 88, respectively; the amino acid sequence of SEQ ID NOS:116 and 119, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:116 and 119, respectively; the amino acid sequence of SEQ ID NOS:125 and 127, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:125 and 127, respectively; the amino acid sequence of SEQ ID NOS:131 and 132, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:131 and 132, respectively.

[0274] In some embodiments, the VH and VL regions of the antibody or antigen-binding fragment thereof, in the provided CAR, comprises: the amino acid sequence of SEQ ID NOS:32 and 33, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:32 and 33, respectively. In some embodiments, the VH and VL regions of the antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NOS:52 and 53, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:52 and 53, respectively. In some embodiments, the VH and VL regions of the antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NOS:61 and 62, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:61 and 62, respectively. In some embodiments, the VH and VL regions of the antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NOS:85 and 88, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:85 and 88, respectively. In some embodiments, the VHand VLregions of the antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NOS:116 and 119, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:116 and 119, respectively. In some embodiments, the VHand VLregions of the antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NOS:125 and 127, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:125 and 127, respectively. In some embodiments, the VHand VLregions of the antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NOS:131 and 132, respectively, or a sequence of amino acids having at least 90% identity to SEQ ID NOS:131 and 132, respectively.

[0275] In some embodiments, in the provided CAR, the antibody or antigen-binding fragment thereof comprises a VH and a VL region, and the VH region comprises a heavy chain complementarity determining region 1 (CDR-H1), a heavy chain complementarity determining region 2 (CDR-H2) and a heavy chain complementarity determining region 3 (CDR-H3) contained within the VH region amino acid sequence selected from any one of SEQ ID NOs: 32, 52, 61, 85, 116, 125, 131; and the VL region comprises a light chain complementarity determining region 1 (CDR-L1), a light chain complementarity determining region 2 (CDR-L2) and a light chain complementarity determining region 3 (CDR-L3) contained within the VL region amino acid sequence selected from any one of SEQ ID NOs: 33, 53, 62, 88, 119, 127, 132.

[0276] In some embodiments, in the provided CAR, the antibody or antigen-binding fragment thereof comprises a VH and a VL region, and the VH region comprises a CDR-H1, a CDR-H2 and a CDR- H3 contained within the amino acid sequence of SEQ ID NO:32, and the VL region comprises a CDR-L1, a CDR-L2 and a CDR-L3 contained within the amino acid sequence of SEQ ID NO:33; the VH region comprises a CDR-H1, a CDR-H2 and a CDR-H3 contained within the amino acid sequence of SEQ ID NO:52, and the VL region comprises a CDR-L1, a CDR-L2 and a CDR-L3 contained within the amino acid sequence of SEQ ID NO:53; the VH region comprises a CDR-H1, a CDR-H2 and a CDR-H3 contained within the amino acid sequence of SEQ ID NO:61, and the VL region comprises a CDR-L1, a CDR-L2 and a CDR-L3 contained within the amino acid sequence of SEQ ID NO:62; the VH region comprises a CDR-H1, a CDR-H2 and a CDR-H3 contained within the amino acid sequence of SEQ ID NO:85, and the VL region comprises a CDR-L1, a CDR-L2 and a CDR-L3 contained within the amino acid sequence of SEQ ID NO:88; the VHregion comprises a CDR-H1, a CDR-H2 and a CDR-H3 contained within the amino acid sequence of SEQ ID NO:116, and the VLregion comprises a CDR-L1, a CDR-L2 and a CDR-L3 contained within the amino acid sequence of SEQ ID NO:119; the VHregion comprises a CDR-H1, a CDR-H2 and a CDR-H3 contained within the amino acid sequence of SEQ ID NO:125, and the VLregion comprises a CDR-L1, a CDR-L2 and a CDR-L3 contained within the amino acid sequence of SEQ ID NO:127; the VHregion comprises a CDR-H1, a CDR-H2 and a CDR-H3 contained within the amino acid sequence of SEQ ID NO:131, and the VLregion comprises a CDR-L1, a CDR-L2 and a CDR-L3 contained within the amino acid sequence of SEQ ID NO:132;

[0277] In some embodiments, the VHand VLregions of the antibody or antigen-binding fragment thereof, in the provided CAR, comprises: the amino acid sequence of SEQ ID NOS:32 and 33, respectively. In some embodiments, the VHand VLregions of the antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NOS:52 and 53, respectively. In someembodiments, the VHand VLregions of the antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NOS:61 and 62, respectively. In some embodiments, the VHand VLregions of the antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NOS:85 and 88, respectively. In some embodiments, the VHand VLregions of the antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NOS:116 and 119, respectively. In some embodiments, the VHand VLregions of the antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NOS:125 and 127, respectively. In some embodiments, the VHand VLregions of the antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NOS:131 and 132, respectively.

[0278] In some embodiments, the VH and VL regions of the antibody or antigen-binding fragment thereof provided therein comprise the amino acid sequences selected from: SEQ ID NOS:116 and 119, or any antibody or antigen-binding fragment thereof that has at least 90% sequence identity to any of the above VH and VL, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, or any antibody or antigen-binding fragment thereof that comprises a CDR-H1, CDR-H2 and CDR-H3 contained within the VH region and a CDR-L1, CDR-L2 and CDR-L3 contained within the VL region of any of the above VH and VL.

[0279] 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). In some embodiments, the antibody or antigen-binding fragment is a single domain antibody comprising only the VH region. In some embodiments, the antibody or antigen binding fragment is an scFv comprising a heavy chain variable (VH) region and a light chain variable (VL) region. In some embodiments, the single-chain antibody fragment (e.g. scFv) includes one or more linkers joining two antibody domains or regions, such as a heavy chain variable (VH) region and a light chain variable (VL) region. The linker typically is a peptide linker, e.g., a flexible and / or soluble peptide linker. Among the linkers are those rich in glycine and serine and / or in some cases threonine. In some embodiments, the linkers further include charged residues such as lysine and / or glutamate, which can improve solubility. In some embodiments, the linkers further include one or more proline.

[0280] Accordingly, the provided anti-BCMA antibodies include single-chain antibody fragments, such as scFvs and diabodies, particularly human single-chain antibody fragments, typically comprising linker(s) joining two antibody domains or regions, such VHand VLregions. The linker typically is a peptide linker, e.g., a flexible and / or soluble peptide linker, such as one rich in glycine and serine.

[0281] In some aspects, the linkers rich in glycine and serine (and / or threonine) include at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% such amino acid(s). In some embodiments, they include at least at or about 50%, 55%, 60%, 70%, or 75%, glycine, serine, and / or threonine. In some embodiments, the linker is comprised substantially entirely of glycine, serine, and / orthreonine. The linkers generally are between about 5 and about 50 amino acids in length, typically between at or about 10 and at 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:7) or GGGS (3GS; SEQ ID NO:2), such as between 2, 3, 4, and 5 repeats of such a sequence. Exemplary linkers include those having or consisting of an sequence set forth in SEQ ID NO:1 (GGGGSGGGGSGGGGS). Exemplary linkers further include those having or consisting of the sequence set forth in SEQ ID NO:176 (GSTSGSGKPGSGEGSTKG). Exemplary linkers further include those having or consisting of the sequence set forth in SEQ ID NO:255 (SRGGGGSGGGGSGGGGSLEMA).

[0282] Accordingly, in some embodiments, the provided embodiments include single-chain antibody fragments, e.g., scFvs, comprising one or more of the aforementioned linkers, such as glycine / serine rich linkers, including linkers having repeats of GGGS (SEQ ID NO: 2) or GGGGS (SEQ ID NO: 7), such as the linker set forth in SEQ ID NO:1.

[0283] In some embodiments, the linker has an amino acid sequence containing the sequence set forth in SEQ ID NO:1. The fragment, e.g., scFv, may include a VH region or portion thereof, followed by the linker, followed by a VL region or portion thereof. The fragment, e.g., the scFv, may include the VL region or portion thereof, followed by the linker, followed by the VH region or portion thereof.

[0284] Table 7 provides the SEQ ID NOS: of exemplary antigen-binding domains, such as antibodies or antigen-binding fragments, that can be comprised in the provided BCMA-binding receptors, such as anti-BCMA chimeric antigen receptors (CARs). In some embodiments, the BCMA-binding receptor contains a BCMA-binding antibody or fragment thereof, comprising a VHregion that comprises the CDR-H1, CDR-H2, and CDR-H3 sequence and a VLregion that comprises the CDR-L1, CDR-L2 and CDR-L3 sequence set forth in the SEQ ID NOS: listed in each row of Table 7 below (by Kabat numbering). In some embodiments, the BCMA-binding receptor contains a BCMA-binding antibody or fragment thereof, comprising a VHregion sequence and a VLregion sequence set forth in the SEQ ID NOS: listed in each row of Table 7 below, or an antibody comprising a VHand VLregion amino acid sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the VHregion sequence and the VLregion sequence set forth in the SEQ ID NOS: listed in each row of Table 7 below. In some embodiments, the BCMA-binding receptor contains a BCMA-binding antibody or fragment thereof, comprising a VHregion sequence and a VLregion sequence set forth in the SEQ ID NOS: listed in each row of Table 7 below. In some embodiments, the BCMA-binding receptor contains a BCMA-binding antibody or fragment thereof, comprising an scFv sequence set forth in the SEQ ID NOS: listed in each row of Table 7 below, or an antibody comprising an scFv amino acid sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the scFvsequence set forth in the SEQ ID NOS: listed in each row of Table 7 below. In some embodiments, the BCMA-binding receptor contains a BCMA-binding antibody or fragment thereof, comprising an scFv sequence set forth in SEQ ID NO:114 or an antibody comprising an scFv amino acid sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In some embodiments, the BCMA-binding receptor contains a BCMA-binding antibody or fragment thereof, comprising an scFv sequence set forth in the SEQ ID NOS: listed in each row of Table 7 below. In some embodiments, the BCMA-binding receptor contains a BCMA-binding antibody or fragment thereof, comprising an scFv sequence set forth in SEQ ID NO: 114.

[0285] Among the antibodies, e.g. antigen-binding fragments, in the provided CARs, are human antibodies. In some embodiments of a provided human anti-BCMA antibody, e.g., antigen-binding fragments, the human antibody contains a VH region that comprises a portion having at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence encoded by a germline nucleotide human heavy chain V segment, a portion having at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence encoded by a germline nucleotide human heavy chain D segment, and / or a portion having at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence encoded by a germline nucleotide human heavy chain J segment; and / or contains a VLregion that comprises a portion having at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence encoded by a germline nucleotide human kappa or lambda chain V segment, and / or a portion having at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence encoded by a germline nucleotide human kappa or lambda chain J segment. In some embodiments, the portion of the VHregion corresponds to the CDR-H1, CDR-H2 and / or CDR-H3. In some embodiments, the portion of the VHregion corresponds to the framework region 1 (FR1), FR2, FR2 and / or FR4. In some embodiments, the portion of the VLregion corresponds to the CDR-L1, CDR-L2 and / or CDR-L3. In some embodiments, the portion of the VLregion corresponds to the FR1, FR2, FR2 and / or FR4.

[0286] In some embodiments, the human antibody, e.g., antigen-binding fragment, contains a CDR- H1 having at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence of the corresponding CDR-H1 region within a sequence encoded by a germline nucleotide human heavy chain V segment. For example, the human antibody in some embodiments contains a CDR-H1 having a sequence that is 100% identical or with no more than one, two or three amino acid differences as compared to the corresponding CDR-H1 region within a sequence encoded by a germline nucleotide human heavy chain V segment.

[0287] In some embodiments, the human antibody, e.g., antigen-binding fragment, contains a CDR- H2 having at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence of the corresponding CDR-H2 region within a sequence encoded by a germline nucleotide human heavy chain V segment. For example, the human antibody in some embodiments contains a CDR-H2 having a sequence that is 100% identical or with no more than one, two or three amino acid difference as compared to the corresponding CDR-H2 region within a sequence encoded by a germline nucleotide human heavy chain V segment.

[0288] In some embodiments, the human antibody, e.g., antigen-binding fragment, contains a CDR- H3 having at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence of the corresponding CDR-H3 region within a sequence encoded by a germline nucleotide human heavy chain V segment, D segment and J segment. For example, the human antibody in some embodiments contains a CDR-H3 having a sequence that is 100% identical or with no more than one, two or three amino acid differences as compared to the corresponding CDR-H3 region within a sequence encoded by a germline nucleotide human heavy chain V segment, D segment and J segment.

[0289] In some embodiments, the human antibody, e.g., antigen-binding fragment, contains a CDR- L1 having at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence of the corresponding CDR-L1 region within a sequence encoded by a germline nucleotide human light chain V segment. For example, the human antibody in some embodiments contains a CDR-L1 having a sequence that is 100% identical or with no more than one, two or three amino acid differences as compared to the corresponding CDR-L1 region within a sequence encoded by a germline nucleotide human light chain V segment.

[0290] In some embodiments, the human antibody, e.g., antigen-binding fragment, contains a CDR- L2 having at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence of the corresponding CDR-L2 region within a sequence encoded by a germline nucleotide human light chain V segment. For example, the human antibody in some embodiments contains a CDR-L2 having a sequence that is 100% identical or with no more than one, two or three amino acid difference as compared to the corresponding CDR-L2 region within a sequence encoded by a germline nucleotide human light chain V segment.

[0291] In some embodiments, the human antibody, e.g., antigen-binding fragment, contains a CDR- L3 having at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an amino acid sequence of the corresponding CDR-L3 region within a sequence encoded by a germline nucleotide human light chain V segment and J segment. For example, the human antibody in some embodiments contains a CDR-L3 having a sequence that is 100% identical or with no more than one, two or three amino acid differences as compared to the corresponding CDR-L3 region within a sequence encoded by a germline nucleotide human light chain V segment and J segment.

[0292] In some embodiments, the human antibody, e.g., antigen-binding fragment, contains a framework region that contains human germline gene segment sequences. For example, in some embodiments, the human antibody contains a VH region in which the framework region, e.g. FR1, FR2, FR3 and FR4, has at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a framework region encoded by a human germline antibody segment, such as a V segment and / or J segment. In some embodiments, the human antibody contains a VL region in which the framework region e.g. FR1, FR2, FR3 and FR4, has at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a framework region encoded by a human germline antibody segment, such as a V segment and / or J segment. For example, in some such embodiments, the framework region sequence contained within the VH region and / or VL region differs by no more than 10 amino acids, such as no more than 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid, compared to the framework region sequence encoded by a human germline antibody segment.

[0293] In some embodiments, the reference antibody can be a mouse anti-BCMA scFv described in International Patent App. Pub. No. WO 2010 / 104949.

[0294] The antibody, e.g., antigen-binding fragment, may contain at least a portion of an immunoglobulin constant region, such as one or more constant region domain. In some embodiments, the constant regions include a light chain constant region and / or a heavy chain constant region 1 (CH1). In some embodiments, the antibody includes a CH2 and / or CH3 domain, such as an Fc region. In some embodiments, the Fc region is an Fc region of a human IgG, such as an IgG1 or IgG4. 2. Spacer

[0295] In some embodiments, the recombinant receptor such as a CAR comprising an antibody (e.g., antigen-binding fragment) provided herein, such as those expressed by engineered cells employed in the methods and uses provided herein, further includes a spacer or spacer region. The spacer typically is a polypeptide spacer and in general is located within the CAR between the antigen binding domain and the transmembrane domain of the CAR. In some aspects, the spacer may be or include at least a portion of an immunoglobulin constant region or variant or modified version thereof, such as a hinge region of an immunoglobulin, such as an IgG hinge region, e.g., an IgG4 or IgG4-derived hinge region, and / or a CH1 / CL and / or Fc region. In some embodiments, the constant region or one or more of the portion(s)thereof is of a human IgG, such as of a human IgG4 or IgG1 or IgG2. In general, the spacer, such as the portion of the constant region, serves as a spacer region between the antigen-recognition component (e.g., scFv) and transmembrane domain. In some embodiments, the length and / or composition of the spacer is designed to optimize or promote certain features of the interaction between the CAR and its target; in some aspects, it is designed to optimize the biophysical synapse distance between the CAR-expressing cell and the cell expressing the target of the CAR during or upon or following binding of the CAR to its target on the target-expressing cell; in some aspects, the target expressing cell is a BCMA-expressing tumor cell. In some embodiments, The CAR is expressed by a T-cell, and the length of the spacer is of a length that is compatible for T-cell activation or to optimize CAR T-cell performance. In some embodiments, the spacer is a spacer region, located between the ligand-binding domain and the transmembrane domain, of the recombinant receptor, e.g., CAR. In some embodiments, the spacer region is a region located between the ligand-binding domain and the transmembrane domain, of the recombinant receptor, e.g., CAR.

[0296] In some embodiments, the spacer can be of a length that provides for increased responsiveness of the cell following antigen binding, as compared to in the absence of the spacer and / or in the presence of a different spacer, such as one different only in length. In some embodiments, the spacer is at least 100 amino acids in length, such as at least 110, 125, 130, 135, 140, 145, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, or 250 amino acids in length. In some examples, the spacer is at or about 12 amino acids in length or is no more than 12 amino acids in length. Exemplary spacers include those having at least about 10 to 300 amino acids, about 10 to 200 amino acids, about 50 to 175 amino acids, about 50 to 150 amino acids, about 10 to 125 amino acids, about 50 to 100 amino acids, about 100 to 300 amino acids, about 100 to 250 amino acids, about 125 to 250 amino acids, or about 200 to 250 amino acids, and including any integer between the endpoints of any of the listed ranges. In some embodiments, a spacer or a spacer region is at least about 12 amino acids, at least about 119 amino acids or less, at least about 125 amino acids, at least about 200 amino acids, or at least about 220 amino acids, or at least about 225 amino acids in length.

[0297] In some embodiments, the spacer has a length of 125 to 300 amino acids in length, 125 to 250 amino acids in length, 125 to 230 amino acids in length, 125 to 200 amino acids in length, 125 to 180 amino acids in length, 125 to 150 amino acids in length, 150 to 300 amino acids in length, 150 to 250 amino acids in length, 150 to 230 amino acids in length, 150 to 200 amino acids in length, 150 to 180 amino acids in length, 180 to 300 amino acids in length, 180 to 250 amino acids in length, 180 to 230 amino acids in length, 180 to 200 amino acids in length, 200 to 300 amino acids in length, 200 to 250 amino acids in length, 200 to 230 amino acids in length, 230 to 300 amino acids in length, 230 to 250 amino acids in length or 250 to 300 amino acids in length. In some embodiments, the spacer is at least orat least about or is or is about 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 221, 222, 223, 224, 225, 226, 227, 228 or 229 amino acids in length, or a length between any of the foregoing.

[0298] Exemplary spacers include those containing portion(s) of an immunoglobulin constant region such as those containing an Ig hinge, such as an IgG hinge domain. In some aspects, the spacer includes an IgG hinge alone, an IgG hinge linked to one or more of a CH2 and CH3 domain, or IgG hinge linked to the CH3 domain. In some embodiments, the IgG hinge, CH2 and / or CH3 can be derived all or in part from IgG4 or IgG2. In some embodiments, the spacer can be a chimeric polypeptide containing one or more of a hinge, CH2 and / or CH3 sequence(s) derived from IgG4, IgG2, and / or IgG2 and IgG4. In some embodiments, the hinge region comprises all or a portion of an IgG4 hinge region and / or of an IgG2 hinge region, wherein the IgG4 hinge region is optionally a human IgG4 hinge region and the IgG2 hinge region is optionally a human IgG2 hinge region; the CH2 region comprises all or a portion of an IgG4 CH2 region and / or of an IgG2 CH2 region, wherein the IgG4 CH2 region is optionally a human IgG4 CH2 region and the IgG2 CH2 region is optionally a human IgG2 CH2 region; and / or the CH3 region comprises all or a portion of an IgG4 CH3 region and / or of an IgG2 CH3 region, wherein the IgG4 CH3 region is optionally a human IgG4 CH3 region and the IgG2 CH3 region is optionally a human IgG2 CH3 region. In some embodiments, the hinge, CH2 and CH3 comprises all or a portion of each of a hinge region, CH2 and CH3 from IgG4. In some embodiments, the hinge region is chimeric and comprises a hinge region from human IgG4 and human IgG2; the CH2 region is chimeric and comprises a CH2 region from human IgG4 and human IgG2; and / or the CH3 region is chimeric and comprises a CH3 region from human IgG4 and human IgG2. In some embodiments, the spacer compri...

Claims

CLAIMS 1. A method of treating a multiple myeloma (MM), the method comprising administering to a subject having or suspected of having a MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein: the composition comprises CD8+T cells expressing the CAR and CD4+T cells expressing the CAR; the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 80 × 106CAR-expressing T cells, inclusive; at least or at least about 80% of the cells in the composition are CD3+cells; and at least or at least about 80% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype.

2. A method of treating a multiple myeloma (MM), the method comprising administering to a subject having or suspected of having a MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets B cell maturation antigen (BCMA), wherein: the composition comprises CD8+T cells expressing the CAR and CD4+T cells expressing the CAR; the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 100 × 106CAR-expressing T cells, inclusive; at least or at least about 80% of the cells in the composition are CD3+cells; and at least or at least about 50% of the CD4+CAR+T cells in the composition are CD27+CCR7+and / or at least or at least about 50% of the CD8+CAR+T cells in the composition are CD27+CCR7+.

3. The method of claim 1 or claim 2, wherein the composition comprises CD4+T cells expressing the CAR and CD8+T cells expressing the CAR at a ratio between about 1:2.5 and about 5:

1.

4. The method of any of claims 1-3, wherein the composition comprises CD4+T cells expressing the CAR and CD8+T cells expressing the CAR at a ratio between about 1:2 and about 4:1, between about 1:1.5 and about 2:1, or at or at about 1:

1.

5. The method of any of claims 1-3, wherein the composition comprises CD4+T cells expressing the CAR and CD8+T cells expressing the CAR at a ratio between about 5:1 and about 2:1, between about 4:1 and about 2:1, between about 3:1 and about 2:1, at or at about 5:1, at or at about 4:1, at or at about 3:1, or at or at about 2:1.

6. The method of any of claims 2-5, wherein the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 80 × 106CAR-expressing T cells, inclusive.

7. The method of any of claims 1-6, wherein the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 40 × 106CAR-expressing T cells, inclusive.

8. The method of any of claims 1-7, wherein the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 20 × 106CAR-expressing T cells, inclusive.

9. The method of any of claims 1-8, wherein the composition comprises between at or about 5 × 106CAR-expressing T cells and at or about 10 × 106CAR-expressing T cells, inclusive.

10. The method of any of claims 1-8, wherein the composition comprises between at or about 10 × 106CAR-expressing T cells and at or about 20 × 106CAR-expressing T cells, inclusive.

11. The method of any of claims 1-8 and 10, wherein the composition comprises at or about 20 × 106CAR-expressing T cells.

12. The method of any of claims 1-7, wherein the composition comprises at or about 30 × 106CAR-expressing T cells.

13. The method of any of claims 1-7, wherein the composition comprises at or about 40 × 106CAR-expressing T cells.

14. The method of any of claims 1-13, wherein at least or at least about 90% of the cells in the composition are CD3+cells.

15. The method of any of claims 1-14, wherein at least or at least about 91%, at least or at least about 92%, at least or at least about 93%, at least or at least about 94%, at least or at least about 95%, or at least or at least about 96% of the cells in the composition are CD3+cells.

16. The method of any of claims 1-15, wherein between at or about 2% and at or about 30% of the CAR+T cells in the composition express a marker of apoptosis, optionally Annexin V or active Caspase 3.

17. The method of any of claims 1-16, wherein between at or about 5% and at or about 10% of the CAR+T cells in the composition express a marker of apoptosis, optionally Annexin V or active Caspase 3.

18. The method of any of claims 1-16, wherein between at or about 10% and at or about 15% of the CAR+T cells in the composition express a marker of apoptosis, optionally Annexin V or active Caspase 3.

19. The method of any of claims 1-16, wherein between at or about 15% and at or about 20% of the CAR+T cells in the composition express a marker of apoptosis, optionally Annexin V or active Caspase 3.

20. The method of any of claims 1-16, wherein between at or about 20% and at or about 30% of the CAR+T cells in the composition express a marker of apoptosis, optionally Annexin V or active Caspase 3.

21. The method of any of claims 1-16, wherein at or about 5%, at or about 10%, at or about 15%, at or about 20%, at or about 25%, or at or about 30% of the CAR+T cells in the composition express a marker of apoptosis, optionally Annexin V or active Caspase 3.

22. The method of any of claims 2-21, wherein at least or at least about 80% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype.

23. The method of any of claims 1-22, wherein between at or about 80% and at or about 85% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype.

24. The method of any of claims 1-22, wherein between at or about 85% and at or about 90% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype.

25. The method of any of claims 1-22, wherein between at or about 90% and at or about 95% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype.

26. The method of any of claims 1-22, wherein between at or about 95% and at or about 99% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype.

27. The method of any of claims 1-22, wherein at or about 85%, at or about 90%, at or about 95%, or at or about 99% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype.

28. The method of any of claims 1 and 3-27, wherein the at least or at least about 80% of the CAR+T cells in the composition that are of a naïve-like or central memory phenotype are surface positive for a marker expressed on naïve-like or central memory T cells.

29. The method of claim 28, wherein the marker expressed on naïve-like or central memory T cell is selected from the group consisting of CD45RA, CD27, CD28, and CCR7.

30. The method of any of claims 1 and 3-29, wherein the at least or at least about 80% of the CAR+T cells in the composition that are of a naïve-like or central memory phenotype have a phenotype selected from CCR7+CD45RA+, CD27+CCR7+, or CD62L-CCR7+.

31. The method of any of claims 1-30, wherein between at or about 80% and at or about 85%, between at or about 85% and at or about 90%, between at or about 90% and at or about 95%, between at or about 95% and at or about 99% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype selected from CCR7+CD45RA+, CD27+CCR7+, or CD62L-CCR7+.

32. The method of any of claims 1-31, wherein at or about 80%, at or about 85%, at or about 90%, at or about 95%, or at or about 99% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype selected from CCR7+CD45RA+, CD27+CCR7+, or CD62L-CCR7+.

33. The method of any of claims 1-32, wherein at or about 80%, at or about 85%, at or about 90%, at or about 95%, or at or about 99% of the CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

34. The method of any of claims 1-33, wherein at least or at least about 50% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-.

35. The method of any of claims 1-34, wherein at least or at least about 60% of the CD4+CAR+T cells in the composition are of a naïve –like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-.

36. The method of any of claims 1-35, wherein at least or at least about 70% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-.

37. The method of any of claims 1-36, wherein at least or at least about 80% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-.

38. The method of any of claims 1-37, wherein at least or at least about 85% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-.

39. The method of any of claims 1-38, wherein at least or at least about 50% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

40. The method of any of claims 1-39, wherein at least or at least about 60% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

41. The method of any of claims 1-40, wherein at least or at least about 70% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

42. The method of any of claims 1-41, wherein at least or at least about 80% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

43. The method of any of claims 1-42, wherein at least or at least about 85% of the CD4+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

44. The method of any of claims 1-43, wherein at least or at least about 50% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-.

45. The method of any of claims 1-44, wherein at least or at least about 60% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-.

46. The method of any of claims 1-45, wherein at least or at least about 70% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-.

47. The method of any of claims 1-46, wherein at least or at least about 80% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-.

48. The method of any of claims 1-47, wherein at least or at least about 85% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CCR7+CD45RA+or CCR7+CD45RA-.

49. The method of any of claims 1-48, wherein at least or at least about 50% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

50. The method of any of claims 1-49, wherein at least or at least about 60% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

51. The method of any of claims 1-50, wherein at least or at least about 70% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

52. The method of any of claims 1-51, wherein at least or at least about 80% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

53. The method of any of claims 1-52, wherein at least or at least about 85% of the CD8+CAR+T cells in the composition are of a naïve-like or central memory phenotype that is CD27+CCR7+.

54. The method of any of claims 1-53, wherein the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is less than or less than about 0.

9.

55. The method of any of claims 1-54, wherein the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is between at or about 0.9 and at or about 0.

8.

56. The method of any of claims 1-54, wherein the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is less than or less than about 0.

8.

57. The method of any of claims 1-54 and 56, wherein the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is between at or about 0.8 and at or about 0.

7.

58. The method of any of claims 1-54 and 56, wherein the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is between at or about 0.7 and at or about 0.6.

59. The method of any of claims 1-54 and 56, wherein the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is between at or about 0.6 and at or about 0.

5.

60. The method of any of claims 1-54 and 56, wherein the fraction of integrated vector copy number (iVCN) to total VCN in the CAR+T cells in the composition, on average, is between at or about 0.5 and at or about 0.

4.

61. The method of any of claims 1-60, wherein the integrated vector copy number (iVCN) of the CAR+T cells in the composition, on average, is between or between about 0.4 copies per diploid genome and 2.0 copies per diploid genome, inclusive.

62. The method of any of claims 1-61, wherein the integrated vector copy number (iVCN) of the CAR+T cells in the composition, on average, is between or between about 0.8 copies per diploid genome and 2.0 copies per diploid genome, inclusive.

63. The method of any of claims 1-62, wherein the integrated vector copy number (iVCN) of the CAR+T cells in the composition, on average, is between or between about 0.8 copies per diploid genome and 1.0 copies per diploid genome, inclusive.

64. The method of any of claims 1-62, wherein the integrated vector copy number (iVCN) of the CAR+T cells in the composition, on average, is between or between about 1.0 copies per diploid genome and 1.5 copies per diploid genome, inclusive.

65. The method of any of claims 1-62, wherein the integrated vector copy number (iVCN) of the CAR+T cells in the composition, on average, is between or between about 1.5 copies per diploid genome and 2.0 copies per diploid genome, inclusive.

66. The method of any of claims 1-65, wherein at or prior to the administration of the composition comprising engineered T cells, the subject has received at least 3 prior antimyeloma treatment regimens.

67. The method of any of claims 1-66, wherein at or prior to the administration of the composition comprising engineered T cells, the subject has received all three of the following antimyeloma treatment regimens: autologous stem cell transplant (ASCT); a regimen comprising an immunomodulatory agent and a proteasome inhibitor; and an anti-CD38 agent.

68. The method of claim 66 or claim 67, wherein at or prior to the administration of the composition comprising engineered T cells, the subject is refractory to the last antimyeloma treatment regimen.

69. The method of claim 68, wherein refractory myeloma is defined as documented progressive disease during or within 12 months, measured from the last dose, of completing treatment with the last anti-myeloma treatment regimen.

70. The method of any of claims 1-69, wherein the subject has not received a prior CAR T cell or genetically-modified T cell therapy.

71. The method of any of claims 1-70, wherein the subject has not received a prior BCMA- targeted therapy such as an anti-BCMA monoclonal antibody or bispecific antibody.

72. The method of any of claims 1-71, further comprising obtaining a leukapheresis sample from the subject for manufacturing the composition comprising engineered T cells.

73. The method of any one of claims 1-72, wherein the CAR comprises: (a) an extracellular antigen-binding domain, comprising: a variable heavy chain (VH) comprising a heavy chain complementarity determining region 1 (CDR-H1), a heavy chain complementarity determining region 2 (CDR-H2) and a heavy chain complementarity determining region 3 (CDR-H3) contained within the sequence set forth in SEQ ID NO: 116 and a variable light chain (VL) comprising a light chain complementarity determining region 1 (CDR-L1), a light chain complementarity determining region 2 (CDR-L2) and a light chain complementarity determining region 3 (CDR-L3) contained within the sequence set forth in SEQ ID NO: 119; a VH comprising a CDR-H1, a CDR-H2 and a CDR-H3 sequences set forth in SEQ ID NOS:97, 101 and 103, respectively, and a VL comprising a CDR-L1, a CDR-L2 and a CDR-L3 sequences set forth in SEQ ID NOS:105, 107 and 108, respectively; a VH comprising a CDR-H1, a CDR-H2 and a CDR-H3 sequences set forth in SEQ ID NOS:96, 100 and 103, respectively, and a VL comprising a CDR-L1, a CDR-L2 and a CDR-L3 sequences set forth in SEQ ID NOS:105, 107 and 108, respectively; a VH comprising a CDR-H1, a CDR-H2 and a CDR-H3 sequences set forth in SEQ ID NOS:95, 99 and 103, respectively, and a VLcomprising a CDR-L1, a CDR-L2 and a CDR-L3 sequences set forth in SEQ ID NOS: 105, 107 and 108, respectively; a VHcomprising a CDR-H1, a CDR-H2 and a CDR-H3 sequences set forth in SEQ ID NOS:94, 98 and 102, respectively, and a VLcomprising a CDR-L1, a CDR-L2 and a CDR-L3 sequences set forthin SEQ ID NOS: 104, 106 and 108, respectively; or a VHcomprising the amino acid sequence of SEQ ID NO: 116 and a VLcomprising the amino acid sequence of SEQ ID NO: 119; (b) a spacer comprising an IgG4 / 2 chimeric hinge or a modified IgG4 hinge; an IgG2 / 4 chimeric CH2 region; and an IgG4 CH3 region, which optionally is about 228 amino acids in length; optionally wherein the spacer is set forth in SEQ ID NO: 174; (c) a transmembrane domain, optionally a transmembrane domain from a human CD28; and (d) an intracellular signaling region comprising a cytoplasmic signaling domain of a CD3-zeta (CD3ζ) chain and a costimulatory signaling region comprising an intracellular signaling domain of a T cell costimulatory molecule or a signaling portion thereof.

74. The method of claim 73, wherein the VH is or comprises the amino acid sequence of SEQ ID NO: 116; and the VL is or comprises the amino acid sequence of SEQ ID NO:

119.

75. The method of claim 73 or claim 74, wherein the extracellular antigen-binding domain comprises an scFv.

76. The method of any of claims 73-75, wherein the VH and the VL are joined by a flexible linker.

77. The method of claim 75 or claim 76, wherein the scFv comprises a linker comprising the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:1).

78. The method of any of claims 73-77, wherein the extracellular antigen-binding domain comprises the amino acid sequence of SEQ ID NO: 114 or 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 SEQ ID NO:

114.

79. The method of any of claims 73-78, wherein the extracellular antigen-binding domain comprises the amino acid sequence of SEQ ID NO:

114.

80. The method of any of claims 73-79, wherein the cytoplasmic signaling domain is or comprises the sequence set forth in SEQ ID NO:143 or a sequence of amino acids that has at least 90% sequence identity to SEQ ID NO:

143.

81. The method of any of claims 73-80, wherein the costimulatory signaling region comprises an intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof.

82. The method of any of claims 73-81, wherein the costimulatory signaling region comprises an intracellular signaling domain of 4-1BB, optionally human 4-1BB.

83. The method of any of claims 73-82, wherein the costimulatory signaling region is or comprises the sequence set forth in SEQ ID NO:4 or a sequence of amino acids that has at least 90% sequence identity to the sequence set forth in SEQ ID NO:

4.

84. The method of any of claims 73-83, wherein the costimulatory signaling region is between the transmembrane domain and the cytoplasmic signaling domain of a CD3-zeta (CD3ζ) chain.

85. The method of any of claims 73-84, wherein the transmembrane domain is or comprises a transmembrane domain from human CD28.

86. The method of any of claims 73-85, wherein the transmembrane domain is or comprises the sequence set forth in SEQ ID NO:138 or a sequence of amino acids that has at least 90% sequence identity to SEQ ID NO:

138.

87. The method of any of claims 73-86, wherein the CAR comprises from its N to C terminus in order: the extracellular antigen-binding domain, the spacer, the transmembrane domain, and the intracellular signaling region.

88. The method of any of claims 1-87, wherein the CAR comprises: (a) an extracellular antigen-binding domain comprising the sequence set forth in SEQ ID NO: 114 or a sequence of amino acids having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 114; (b) a spacer comprising the sequence set forth in SEQ ID NO: 174 or a sequence of amino acids that has at least 90% sequence identity to SEQ ID NO:174; (c) a transmembrane domain comprising the sequence set forth in SEQ ID NO:138 or a sequence of amino acids that has at least 90% sequence identity to SEQ ID NO:138; and (d) an intracellular signaling region comprising a cytoplasmic signaling comprising the sequence set forth in SEQ ID NO:143 or a sequence of amino acids that has at least 90% sequence identity to SEQ ID NO:143 and a costimulatory signaling region comprising the sequence set forth in SEQ ID NO:4 or a sequence of amino acids that has at least 90% sequence identity to the sequence set forth in SEQ ID NO:

4.

89. The method of any of claims 1-88, wherein the CAR comprises: (a) an extracellular antigen-binding domain, comprising the sequence set forth in SEQ ID NO: 114;(b) a spacer comprising the sequence set forth in SEQ ID NO: 174; (c) a transmembrane domain comprising the sequence set forth in SEQ ID NO:138; and (d) an intracellular signaling region comprising a cytoplasmic signaling comprising the sequence set forth in SEQ ID NO:143 and a costimulatory signaling region comprising the sequence set forth in SEQ ID NO:

4.

90. The method of any of claims 1-89, wherein the CAR comprises the sequence set forth in SEQ ID NO:

19.

91. The method of any of claims 1-90, wherein prior to the administration, the subject has received a lymphodepleting therapy comprising the administration of fludarabine at or about 20-40 mg / m2body surface area of the subject, optionally at or about 30 mg / m2, daily, for 2-4 days, and / or cyclophosphamide at or about 200-400 mg / m2body surface area of the subject, optionally at or about 300 mg / m2, daily, for 2-4 days.

92. The method of any of claims 1-91, wherein the method is capable of achieving a specified response or outcome, optionally at a designated timepoint following initiation of the administration, in at least one of or in at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of subjects in a cohort of subjects having the MM, wherein: the response or outcome is selected from the group consisting of objective response (OR), complete response (CR), stringent complete response (sCR), very good partial response (VGPR), partial response (PR) and minimal response (MR); the response or outcome is or comprises an OR; and / or the response or outcome is or comprises a CR.

93. The method of claim 92, wherein the response or outcome is durable for greater than at or about 3, 6, 9 or 12 months.

94. The method of claim 92 or claim 93, wherein the response or outcome determined at or about 3, 6, 9 or 12 months after the designated timepoint is equal to or improved compared to the response or outcome determined at the designated timepoint.

95. The method of any of claims 1-94, wherein the method does not result in any cytokine release syndrome (CRS) in at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM .

96. The method of any of claims 1-95, wherein the method does not result in severe cytokinerelease syndrome (CRS) in at least at least at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM.

97. The method of any of claims 1-96, wherein the method does not result in any neurotoxicity in at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM.

98. The method of any of claims 1-97, wherein the method does not result in severe neurotoxicity in at least at least at least 70%, at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM .

99. The method of any of claims 1-98, wherein the method does not result in severe CRS and severe neurotoxicity in at least at least at least 70%, at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM.

100. The method of claim on any of claims 1-99, wherein the method does not result in severe CRS and severe neurotoxicity in at least 80%, at least 90%, or at least 95% of subjects in the cohort of subjects having the MM.

101. The method of claim 96, 99 and 100, wherein the severe CRS is grade 3 or higher, grade 4 or higher or grade 5 CRS.

102. The method of claim 98, 99 or 100 wherein the severe neurotoxicity is grade 3 or higher, grade 4 or higher or grade 5 CRS.

103. The method of any of claims 1-102, wherein the administration of the composition is carried out on an outpatient basis o and / or without admitting the subject to a hospital and / or without an overnight stay at a hospital and / or without requiring admission to or an overnight stay at a hospital, optionally unless or until the subject exhibits a sustained fever or a fever that is or has not been reduced or not reduced by more than 1ºC after treatment with an antipyretic.

104. The method of any of claims 1-103, wherein the composition comprising engineered T cells is administered parenterally, optionally intravenously.

105. The method of any of claims 1-104, wherein the subject is a human subject.

106. The method of any of claims 1-105, wherein the composition comprising engineered T cells is produced by a manufacturing process comprising: (i) exposing an input composition comprising primary T cells, optionally an input composition comprising autologous T cells selected from the subject, with a stimulatory reagent comprising an oligomeric particle reagent comprising a plurality of streptavidin mutein molecules under conditions to stimulate T cells, thereby generating a stimulated population, wherein: the oligomeric particle reagent comprises a first agent comprising an anti-CD3 antibody or antigen binding fragment thereof and a second agent comprising an anti-CD28 antibody or antigen binding fragment thereof; (ii) introducing into T cells of the stimulated population, a heterologous polynucleotide encoding the CAR that targets BCMA, thereby generating a population of transformed cells; (iii) incubating the population of transformed cells for up to 96 hours; and (iv) harvesting T cells of the population of transformed cells, thereby producing a composition of engineered cells, wherein the harvesting is carried out at a time between 24 and 120 hours, inclusive, after the exposing to the stimulatory reagent is initiated.

107. The method of claim 106, wherein the anti-CD3 antibody or antigen binding fragment is a Fab and the anti-CD28 antibody or antigen binding fragment is a Fab.

108. The method of claim 106 or claim 107, wherein the first agent and the second agent each comprise a streptavidin-binding peptide that reversibly binds the first agent and the second agent to the oligomeric particle reagent, optionally wherein the streptavidin-binding peptide comprises the sequence of amino acids set forth in any of SEQ ID NOS:266-270.

109. The method of any of claims 106-108, wherein the streptavidin mutein molecule is a tetramer of a streptavidin mutein comprising amino acid residues Val44-Thr45-Ala46-Arg47 or Ile44- Gly45-Ala46-Arg47, optionally wherein the streptavidin mutein comprises the sequence set forth in any of SEQ ID NOS: 257, 272, 275, 277, 279, 273 or 276.

110. The method of any of claims 106-109, wherein the oligomeric particle reagent comprises between 1,000 and 5,000 streptavidin mutein tetramers, inclusive.

111. The method of any of claims 106-110, wherein the method further comprises, prior to harvesting the cells, adding biotin or a biotin analog after or during the incubation .

112. The method of any of claims 106-111, wherein the harvesting is carried out at a time between 48 and 120 hours, inclusive, after the exposing to the stimulatory reagent is initiated.

113. The method of any of claims 106-112, wherein the harvesting is carried out at a time when integrated vector is detected in the genome but prior to achieving a stable integrated vector copy number (iVCN) per diploid genome.

114. The method of any of claims 106-113, wherein the harvesting is carried out at a time before the total number of viable cells at the harvesting is more than or more than about three times the number of total viable cells of the stimulated population.

115. The method of any of claims 106-114, wherein the harvesting is carried out at a time when the total number of viable cells at the harvesting is at or about three times, at or about two times, or the same or about the same as the number of total viable cells of the stimulated population.

116. The method of any of claims 106-115, wherein the harvesting is carried out at a time when the percentage of CD27+CCR7+cells is greater than or greater than about 50% among total T cells in the population of transformed cells, total CD3+T cells in the population of transformed cells, total CD4+T cells in the population of transformed cells, or total CD8+T cells, or of CAR-expressing cells thereof, in the population of transformed cells.

117. The method of any of claims 106-116, wherein the harvesting is carried out at a time when the percentage of CD45RA+CCR7+and CD45RA-CCR7+cells is greater than or greater than about 60% among total T cells in the population of transformed cells, total CD3+T cells in the population of transformed cells, total CD4+T cells in the population of transformed cells, or total CD8+T cells, or of CAR-expressing cells thereof, in the population of transformed cells.

118. The method of any of claims 1-117, wherein the cells in the administered composition are produced by a manufacturing process to produce an output composition (i) comprising engineered CD4+ T cells and engineered CD8+ T cells and (ii) exhibiting a predetermined feature, wherein iterations of the manufacturing process produce a plurality of the output compositions, optionally from human biological samples, when carried out among a plurality of different individual subjects, in which the predetermined feature of the output composition among the plurality of output compositions is selected from: the mean percentage of cells of a memory phenotype in the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%; the mean percentage of cells of a central memory phenotype in the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%; the mean percentage of cells that are CD27+, CD28+, CCR7+, CD45RA-, CD45RO+, CD62L+, CD3+, CD95+, granzyme B-, and / or CD127+ in the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%; the mean percentage of cells that are CCR7+ / CD45RA- or CCR7+ / CD45RO+ in the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%; the mean percentage of central memory CD4+ T cells in the engineered CD4+ T cells, optionally CAR+CD4+ T cells, of the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%; the mean percentage of central memory CD8+ T cells in the engineered CD8+ T cells, optionally CAR+CD8+ T cells, of the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%; and / or the mean percentage of central memory T cells, optionally CD4+ central memory T cells and CD8+ central memory T cells, in the engineered T cells, optionally CAR+ T cells, of the plurality of the output compositions is between about 40% and about 65%, between about 40% and about 45%, between about 45% and about 50%, between about 50% and about 55%, between about 55% and about 60%, or between about 60% and about 65%.

119. The method of any of claims 1-118, wherein the administered composition is produced by a manufacturing process to produce an output composition exhibiting a predetermined feature, optionally a threshold number of cells expressing the CAR in the output composition, in at least about 80%, about 90%, about 95%, about 97%, about 99%, about 100%, or is 100% of the human biological samples in which it is carried out among a plurality of different individual subjects.

120. The method of any of claims 1-119, wherein the MM is a relapsed and / or refractory multiple myeloma (r / r MM).

121. An article of manufacture comprising a composition comprising genetically engineered cells expressing a chimeric antigen receptor (CAR) that targets BCMA, and instructions foradministering the composition of the cells in accordance with the method of any of claims 1-120.

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