BCMA-targeted chimeric antigen receptor T cell composition, method and use thereof

Abstract

The present invention provides cell compositions for treating subjects with multiple myeloma (MM), as well as related methods. [Solution] A method for treating multiple myeloma (MM) comprising administering to a subject having or having 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 expressing the CAR. + T cells and CD4 expressing the CAR + The composition contains T cells, and the values ​​of both ends are 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~80 × 10⁶ 6 1 or approximately 80 x 10 6 The method comprises CAR-expressing T cells, wherein at least 80% or at least about 80% of the cells in the composition are CD3+ cells, and at least 80% or at least about 80% of the CAR+ T cells in the composition are naive-like or central memory phenotype cells.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority under U.S. Provisional Application No. 62 / 975,731, “BCMA-DIRECTED CHIMERIC ANTIGEN RECEPTOR T CELL COMPOSITIONS AND METHODS AND USES THEREOF,” filed on 12 February 2020, the contents of which are incorporated herein by reference in their entirety.

[0002] Inclusion by referencing sequence listings This application is filed together with an electronic sequence listing. The sequence listing is provided as a file named 735042023540SEQLIST.txt, created on February 10, 2021, and its size is 184 kilobytes. The electronic information of the sequence listing is incorporated in its entirety by reference herein.

[0003] field This disclosure relates, in several aspects, to adoptive cell therapy involving the administration of cell compositions for treating subjects having diseases and conditions such as multiple myeloma (MM), as well as related methods, compositions, uses, and articles of manufacture. [Background technology]

[0004] background Various immunotherapies and / or cell therapies can be utilized to treat diseases and conditions. For example, adoptive cell therapies (including those involving the administration of cells expressing chimeric receptors specific to the disease or disorder of interest, such as chimeric antigen receptors (CARs) and other recombinant antigen receptors, as well as adoptive immunotherapy and adoptive T-cell therapy) may be beneficial in treating cancer or other diseases or disorders. Improved approaches are needed. Methods, uses, and manufactured articles that meet such needs are provided. [Overview of the project]

[0005] overview In one aspect, provided herein is a method of treating multiple myeloma (MM) comprising administering to a subject having or suspected of having 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, and the composition comprises, including the values at both ends, 5×10 6 cells or about 5×10 6 cells of CAR-expressing T cells to 200×10 6 cells or about 200×10 6 cells of CAR-expressing T cells, and at least 80% 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 multiple myeloma (MM) comprising administering to a subject having or suspected of having 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 in a ratio of about 1:2.5 to about 5:1, and the composition comprises, including the values at both ends, 5×10 6 cells or about 5×10 6 cells of CAR-expressing T cells to 200×10 6 cells or about 200×10 6 cells of CAR-expressing T cells, and at least 90% or at least about 90% of the cells in the composition are CD3 + cells.

[0007] In one aspect, this specification provides a method for treating multiple myeloma (MM), comprising the step of administering to a subject having or suspected of having MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets a B cell maturation antigen (BCMA), wherein the composition expresses the CAR. + T cells and CD4 expressing the CAR + The composition contains T cells, and the values ​​of both ends are 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~200 × 10⁶ 6 1 or approximately 200 x 10 6 The composition contains CAR-expressing T cells, and at least 80% or at least about 80% of the cells in the composition are CD3 + Cells and CARs in the composition + A method is provided in which at least 80% or at least about 80% of T cells are naive-like or central memory phenotype cells.

[0008] In one aspect, this specification provides a method for treating multiple myeloma (MM), comprising the step of administering to a subject having or suspected of having MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets a B cell maturation antigen (BCMA), wherein the composition expresses the CAR. + T cells and CD4 expressing the CAR + The composition contains T cells, and the values ​​of both ends are 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~200 × 10⁶ 6 1 or approximately 200 x 10 6 The composition contains CAR-expressing T cells, and at least 80% or at least about 80% of the cells in the composition are CD3 + The cells, and the CD4 in the composition + CAR + At least 50% or at least about 50% of T cells are CD27 + CCR7 + and / or CD8 in the composition + CAR+ At least 50% or at least about 50% of T cells are CD27 + CCR7 + A method is provided that is.

[0009] In one aspect, this specification provides a method for treating multiple myeloma (MM), comprising the step of administering to a subject having or suspected of having MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets a B cell maturation antigen (BCMA), wherein the composition expresses the CAR. + T cells and CD4 expressing the CAR + The composition contains T cells, and the values ​​of both ends are 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~200 × 10⁶ 6 1 or approximately 200 x 10 6 The composition contains CAR-expressing T cells, and at least 80% or at least about 80% of the cells in the composition are CD3 + Cells and CARs in the composition + A method is provided in which the fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, less than 0.9 or approximately less than 0.9.

[0010] In one aspect, this specification provides a method for treating multiple myeloma (MM), comprising the step of administering to a subject having or suspected of having MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets a B cell maturation antigen (BCMA), wherein the composition expresses the CAR. + T cells and CD4 expressing the CAR + The composition contains T cells, and the values ​​of both ends are 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~200 × 10⁶ 6 1 or approximately 200 x 10 6 The composition contains CAR-expressing T cells, and at least 80% or at least about 80% of the cells in the composition are CD3 +Cells and CARs in the composition + A method is provided in which the copy number (iVCN) of the vector into which the T cells are incorporated is, on average, 0.4 copies to 2.0 copies per diploid genome, or approximately 0.4 copies to approximately 2.0 copies per diploid genome, including the values ​​at both ends.

[0011] In some of the arbitrary embodiments, the composition is 50 × 10, including the values ​​at both ends. 6 1 or approximately 50 x 10 6 Individual CAR-expressing T cells ~200 × 10⁶ 6 1 or approximately 200 x 10 6 It contains CAR-expressing T cells. In some of the arbitrary embodiments, the composition has a total of 70 × 10⁶ values, including the values ​​at both ends. 6 1 or approximately 70 x 10 6 Individual CAR-expressing T cells ~200 × 10⁶ 6 1 or approximately 200 x 10 6 It contains CAR-expressing T cells. In some of the arbitrary embodiments, the composition has a total of 80 × 10⁶ values, including the values ​​at both ends. 6 1 or approximately 80 x 10 6 Individual CAR-expressing T cells ~200 × 10⁶ 6 1 or approximately 200 x 10 6 It contains CAR-expressing T cells. In some of the arbitrary embodiments, the composition has a total of 80 × 10⁶ values, including the values ​​at both ends. 6 1 or approximately 80 x 10 6 Individual CAR-expressing T cells ~160 × 10⁶ 6 1 or approximately 160 x 10 6 Contains CAR-expressing T cells.

[0012] In one aspect, this specification provides a method for treating multiple myeloma (MM) in which a subject having or suspected of having MM is administered a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets a B cell maturation antigen (BCMA), wherein the composition is CD4 expressing the CAR. + T cells and CD8 cells expressing the CAR + The composition contains T cells, and the values ​​of both ends are 5 × 10 6one or about 5×10 6 CAR-expressing T cells of ~40×10 6 one or about 40×10 6 CAR-expressing T cells, and at least 80% or at least about 80% of the cells in the composition are CD3 + cells. A method is provided.

[0013] In one aspect, provided herein is a method of treating multiple myeloma (MM) comprising administering to a subject having or suspected of having MM a composition comprising engineered T cells that express 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 in a ratio of about 1:2.5 to about 5:1, and the composition comprises, including the values at both ends, 5×10 6 one or about 5×10 6 CAR-expressing T cells of ~80×10 6 one or about 80×10 6 CAR-expressing T cells, and at least 90% or at least about 90% of the cells in the composition are CD3 + cells. A method is provided.

[0014] In one aspect, provided herein is a method of treating multiple myeloma (MM) comprising administering to a subject having or suspected of having MM a composition comprising engineered T cells that express 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, and the composition comprises, including the values at both ends, 5×10 6 one or about 5×10 6 CAR-expressing T cells of ~10×10 6 one or about 10×10 6 CAR-expressing T cells, and at least 80% or at least about 80% of the cells in the composition are CD3 + cells. A method is provided.

[0015] In one aspect, provided herein is a method of treating multiple myeloma (MM) comprising administering to a subject having or suspected of having 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, and the composition comprises, inclusive of the values at both ends, 5×10 6 cells or about 5×10 6 cells to 80×10 6 cells or about 80×10 6 cells of CAR-expressing T cells, and at least 80% or at least about 80% of the cells in the composition are CD3 + cells, and at least 80% or at least about 80% of the CAR + T cells in the composition are cells of a naive-like or central memory phenotype.

[0016] In one aspect, provided herein is a method of treating multiple myeloma (MM) comprising administering to a subject having or suspected of having 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, and the composition comprises, inclusive of the values at both ends, 5×10 6 cells or about 5×10 6 cells to 100×10 6 cells or about 100×10 6 cells of CAR-expressing T cells, and at least 80% or at least about 80% of the cells in the composition are CD3 + cells, and at least 50% or at least about 50% of the CD4 + CAR + T cells in the composition are CD27 + CCR7 +and / or CD8 in the composition + CAR + At least 50% or at least about 50% of T cells are CD27 + CCR7 + A method is provided that is.

[0017] In one aspect, this specification provides a method for treating multiple myeloma (MM), comprising the step of administering to a subject having or suspected of having MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets a B cell maturation antigen (BCMA), wherein the composition expresses the CAR. + T cells and CD4 expressing the CAR + The composition contains T cells, and the values ​​of both ends are 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~20 × 10⁶ 6 1 or approximately 20 x 10 6 The composition contains CAR-expressing T cells, and at least 80% or at least about 80% of the cells in the composition are CD3 + Cells and CARs in the composition + A method is provided in which at least 80% or at least about 80% of T cells are naive-like or central memory phenotype cells.

[0018] In one aspect, this specification provides a method for treating multiple myeloma (MM), comprising the step of administering to a subject having or suspected of having MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets a B cell maturation antigen (BCMA), wherein the composition expresses the CAR. + T cells and CD4 expressing the CAR + The composition contains T cells, and the values ​​of both ends are 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~80 × 10⁶ 6 1 or approximately 80 x 10 6 The composition contains CAR-expressing T cells, and at least 80% or at least about 80% of the cells in the composition are CD3+ Cells and CARs in the composition + A method is provided in which the fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, less than 0.9 or approximately less than 0.9.

[0019] In one aspect, this specification provides a method for treating multiple myeloma (MM), comprising the step of administering to a subject having or suspected of having MM a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) that targets a B cell maturation antigen (BCMA), wherein the composition expresses the CAR. + T cells and CD4 expressing the CAR + The composition contains T cells, and the values ​​of both ends are 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~80 × 10⁶ 6 1 or approximately 80 x 10 6 The composition contains CAR-expressing T cells, and at least 80% or at least about 80% of the cells in the composition are CD3 + Cells and CARs in the composition + A method is provided in which the copy number (iVCN) of the vector into which the T cells are incorporated is, on average, 0.4 copies to 2.0 copies per diploid genome, or approximately 0.4 copies to approximately 2.0 copies per diploid genome, including the values ​​at both ends.

[0020] In some of the various embodiments, the composition expresses CD4 + T cells and CD8 expressing CAR + The composition contains T cells in a ratio of approximately 1:2.5 to approximately 5:1. In some of the arbitrary embodiments, the composition has a ratio of 5 × 10, including both end values. 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~80 × 10⁶ 6 1 or approximately 80 x 10 6 It contains CAR-expressing T cells. In some of the arbitrary embodiments, the composition has a total of 5 × 10⁶ values, including the values ​​at both ends. 6 pc or approximately 5 x 10 6Individual CAR-expressing T cells ~40 × 10⁶ 6 1 or approximately 40 x 10 6 It contains CAR-expressing T cells. In some of the arbitrary embodiments, the composition has a total of 5 × 10⁶ values, including the values ​​at both ends. 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~20 × 10⁶ 6 1 or approximately 20 x 10 6 Contains CAR-expressing T cells.

[0021] In some of the various embodiments, the composition expresses CD4 + T cells and CD8 expressing CAR + T cells may be present in a ratio of approximately 1:2 to approximately 4:1, approximately 1:1.5 to approximately 2:1, or 1:1 or approximately 1:1. In some of the arbitrary embodiments, the composition may contain CD4 expressing CAR. + T cells and CD8 expressing CAR + T cells may be contained in ratios of approximately 5:1 to approximately 2:1, approximately 4:1 to approximately 2:1, approximately 3:1 to approximately 2:1, 5:1 or approximately 5:1, 4:1 or approximately 4:1, 3:1 or approximately 3:1, or 2:1 or approximately 2:1. In some of the arbitrary embodiments, the composition may contain 5 × 10⁻⁶ values, including both ends. 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~10 × 10 6 1 or approximately 10 x 10 6 It may contain CAR-expressing T cells. In some of the arbitrary embodiments, the composition may contain 10 × 10⁶ values ​​including both ends. 6 1 or approximately 10 x 10 6 Individual CAR-expressing T cells ~20 × 10⁶ 6 1 or approximately 20 x 10 6 It may contain CAR-expressing T cells. In some of the arbitrary embodiments, the composition is 20 × 10 6 1 or approximately 20 x 10 6 It may contain CAR-expressing T cells. In some of the arbitrary embodiments, the composition is 30 × 10 6 1 or approximately 30 x 10 6 It may contain CAR-expressing T cells. In some of the arbitrary embodiments, the composition is 40 × 10 6 1 or approximately 40 x 106 It may contain CAR-expressing T cells. In some of the arbitrary embodiments, the composition is 10 × 10 6 1 or approximately 10 x 10 6 It may contain CAR-expressing T cells. In some of the arbitrary embodiments, the composition is 60 × 10 6 1 or approximately 60 x 10 6 It may contain CAR-expressing T cells. In some of the arbitrary embodiments, the composition is 80 × 10 6 1 or approximately 80 x 10 6 It may contain CAR-expressing T cells. In some of the arbitrary embodiments, the composition is 160 × 10 6 1 or approximately 160 x 10 6 It may contain individual CAR-expressing T cells.

[0022] In some of the various embodiments, at least 90% or at least about 90% of the cells in the composition are CD3 + It is a cell.

[0023] In some of the various embodiments, at least 91% or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, or at least 96% or at least about 96% of the cells in the composition are CD3 + It is a cell. In some of the arbitrary embodiments, CAR in the composition + Of the T cells, 2% or about 2% to 30% or about 30% express apoptosis markers, optionally annexin V or active caspase 3. In some of the arbitrary embodiments, CAR in the composition + Of the T cells, 5% or about 5% to 10% or about 10% express apoptosis markers, optionally annexin V or active caspase 3. In some of the arbitrary embodiments, CAR in the composition +Of the T cells, 10% or about 10%, to 15% or about 15%, express an apoptosis marker, optionally annexin V or active caspase 3. In some of the arbitrary embodiments, CAR in the composition + Of the T cells, 15% or about 15% to 20% or about 20% express apoptosis markers, optionally annexin V or active caspase 3. In some of the arbitrary embodiments, CAR in the composition + Of the T cells, 20% or about 20% to 30% or about 30% express apoptosis markers, optionally annexin V or active caspase 3. In some of the arbitrary embodiments, CAR in the composition + 5% or about 5%, 10% or about 10%, 15% or about 15%, 20% or about 20%, 25% or about 25%, or 30% or about 30% of T cells 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 the arbitrary embodiments, CAR in the composition + At least 80% or at least about 80% of T cells are naive-like or central memory phenotype cells.

[0025] In some of the arbitrary embodiments, CAR in the composition + Of the T cells, 80% or about 80%, to 85% or about 85%, are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, CAR in the composition + Of the T cells, 85% or about 85% to 90% or about 90% are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, CAR in the composition + Of the T cells, 90% or about 90% to 95% or about 95% are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, CAR in the composition+ Of the T cells, 95% or about 95% to 99% or about 99% are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, CAR in the composition + 85% or approximately 85%, 90% or approximately 90%, 95% or approximately 95%, or 99% or approximately 99% of T cells are naive-like or central memory phenotype cells.

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

[0027] In some of the arbitrary embodiments, CAR in a composition which is a naive-like or central memory phenotype cell + At least 80% or at least about 80% of T cells are CCR7 + CD45RA + CD27 + CCR7 + Or CD62L - CCR7 + It has a phenotype selected from the following. In some of the arbitrary embodiments, the composition contains CAR + Of the T cells, 80% or about 80%, 85% or about 85%, 85% or about 85%, 90% or about 90%, 90% or about 90%, 95% or about 95%, 95% or about 95%, and 99% or about 99% are CCR7 + CD45RA + CD27 + CCR7 + Or CD62L - CCR7 +Cells with a naive-like or central memory phenotype selected from the following. In some of the arbitrary embodiments, CAR in the composition + 80% or approximately 80%, 85% or approximately 85%, 90% or approximately 90%, 95% or approximately 95%, or 99% or approximately 99% of T cells are CCR7 + CD45RA + CD27 + CCR7 + Or CD62L - CCR7 + Cells with a naive-like or central memory phenotype selected from the following. In some of the arbitrary embodiments, CAR in the composition + 80% or about 80%, 85% or about 85%, 90% or about 90%, 95% or about 95%, or 99% or about 99% of T cells are CD27 + CCR7 + These are cells with a naive-like or central memory phenotype.

[0028] In some of the embodiments, CD4 in the composition + CAR + At least 50% or at least about 50% of T cells are CCR7 + CD45RA + or CCR7 + CD45RA - These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD4 + CAR + At least 60% or at least about 60% of T cells are CCR7 + CD45RA + or CCR7 + CD45RA - These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD4 + CAR + At least 70% or at least about 70% of T cells are CCR7 + CD45RA + or CCR7 +CD45RA - These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD4 + CAR + At least 80% or at least about 80% of T cells are CCR7 + CD45RA + or CCR7 + CD45RA - These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD4 + CAR + At least 85% or at least about 85% of T cells are CCR7 + CD45RA + or CCR7 + CD45RA - These are cells with a naive-like or central memory phenotype.

[0029] In some of the embodiments, CD4 in the composition + CAR + At least 50% or at least about 50% of T cells are CD27 + CCR7 + These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD4 + CAR + At least 60% or at least about 60% of T cells are CD27 + CCR7 + These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD4 + CAR + At least 70% or at least about 70% of T cells are CD27 + CCR7 + These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD4 + CAR + At least 80% or at least about 80% of T cells are CD27 + CCR7+ These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD4 + CAR + At least 85% or at least about 85% of T cells are CD27 + CCR7 + These are cells with a naive-like or central memory phenotype.

[0030] In some of the embodiments, CD8 in the composition + CAR + At least 50% or at least about 50% of T cells are CCR7 + CD45RA + or CCR7 + CD45RA - These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD8 + CAR + At least 60% or at least about 60% of T cells are CCR7 + CD45RA + or CCR7 + CD45RA - These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD8 + CAR + At least 70% or at least about 70% of T cells are CCR7 + CD45RA + or CCR7 + CD45RA - These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD8 + CAR + At least 80% or at least about 80% of T cells are CCR7 + CD45RA + or CCR7 + CD45RA - These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD8+ CAR + At least 85% or at least about 85% of T cells are CCR7 + CD45RA + or CCR7 + CD45RA - These are cells with a naive-like or central memory phenotype.

[0031] In some of the embodiments, CD8 in the composition + CAR + At least 50% or at least about 50% of T cells are CD27 + CCR7 + These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD8 + CAR + At least 60% or at least about 60% of T cells are CD27 + CCR7 + These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD8 + CAR + At least 70% or at least about 70% of T cells are CD27 + CCR7 + These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD8 + CAR + At least 80% or at least about 80% of T cells are CD27 + CCR7 + These are naive-like or central memory phenotype cells. In some of the arbitrary embodiments, the composition contains CD8 + CAR + At least 85% or at least about 85% of T cells are CD27 + CCR7 + These are cells with a naive-like or central memory phenotype.

[0032] In some of the arbitrary embodiments, CAR in the composition +At least 80% or at least about 80% of the T cells are surface positive for markers expressed on naive-like or central memory T cells. In some of the embodiments, the markers expressed on naive-like or central memory T cells are selected from the group consisting of CD45RA, CD27, CD28, and CCR7. In some of the embodiments, CAR in the composition + At least 80% or at least about 80% of T cells are CCR7 + CD45RA + CD27 + CCR7 + , and / or CD62L - CCR7 + In some of the arbitrary embodiments, CAR in the composition + Of the T cells, 80% or about 80%, 85% or about 85%, 85% or about 85%, 90% or about 90%, 90% or about 90%, 95% or about 95%, 95% or about 95%, and 99% or about 99% are CCR7 + CD45RA + CD27 + CCR7 + , and / or CD62L - CCR7 + In some of the arbitrary embodiments, CAR in the composition + 80% or approximately 80%, 85% or approximately 85%, 90% or approximately 90%, 95% or approximately 95%, or 99% or approximately 99% of T cells are CCR7 + CD45RA + CD27 + CCR7 + , and / or CD62L - CCR7 + In some of the arbitrary embodiments, CAR in the composition + 80% or about 80%, 85% or about 85%, 90% or about 90%, 95% or about 95%, or 99% or about 99% of T cells are CD27 + CCR7 + In some of the various embodiments, CD4 in the composition +CAR + At least 50% or at least about 50% of T cells are CCR7 + CD45RA + or CCR7 + CD45RA - In some of the various embodiments, CD4 in the composition + CAR + At least 60% or at least about 60% of T cells are CCR7 + CD45RA + or CCR7 + CD45RA - That is the case.

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

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

[0035] In some of the arbitrary embodiments, CAR in the composition + The fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, 0.9 or about 0.9 to 0.8 or about 0.8. In some of the arbitrary embodiments, the CAR in the composition + The fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, less than 0.8 or about 0.8. In some of the arbitrary embodiments, the CAR in the composition + The fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, 0.8 or about 0.8 to 0.7 or about 0.7. In some of the arbitrary embodiments, the CAR in the composition + The fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, 0.7 or about 0.7 to 0.6 or about 0.6. In some of the arbitrary embodiments, the CAR in the composition + The fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, 0.6 or about 0.6 to 0.5 or about 0.5. In some of the arbitrary embodiments, the CAR in the composition + The fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, 0.5 or approximately 0.5 to 0.4 or approximately 0.4.

[0036] In some of the arbitrary embodiments, CAR in the composition + The copy number (iVCN) of the vector into which T cells are incorporated is, on average, 0.4 copies to 2.0 copies per diploid genome, or approximately 0.4 copies to approximately 2.0 copies per diploid genome, including the values ​​at both ends.

[0037] In some of the arbitrary embodiments, CAR in the composition +The copy number (iVCN) of the vector into which the T cells are incorporated is, on average, 0.8 copies to 2.0 copies per diploid genome, or approximately 0.8 copies to approximately 2.0 copies per diploid genome, including the values ​​at both ends. In some of the arbitrary embodiments, the CAR in the composition + The copy number (iVCN) of the vector into which the T cells are incorporated is, on average, 0.8 copies to 1.0 copy per diploid genome, or about 0.8 copies to about 1.0 copy per diploid genome, including the values ​​at both ends. In some of the arbitrary embodiments, the CAR in the composition + The copy number (iVCN) of the vector into which the T cells are incorporated is, on average, 1.0 to 1.5 copies per diploid genome, or approximately 1.0 to 1.5 copies per diploid genome, including the values ​​at both ends. In some of the arbitrary embodiments, the CAR in the composition + The copy number (iVCN) of the vector into which T cells are incorporated is, on average, 1.5 to 2.0 copies per diploid genome, or approximately 1.5 to 2.0 copies per diploid genome, including the values ​​at both ends.

[0038] In some of the embodiments, at the time of or prior to administration of the engineered T cell composition, the subject has received at least three prior anti-myeloma treatment regimens. In some of the embodiments, at the time of or prior to administration of the engineered T cell composition, the subject has received three or more treatments selected from autologous stem cell transplantation (ASCT), immunomodulators, proteasome inhibitors, and anti-CD38 agents, optionally four or more prior treatments, unless the subject was not a candidate for one or more of these treatments, or was contraindicated for one or more of these treatments. In some of the embodiments, at the time of or prior to administration of the engineered T cell composition, the subject has optionally received three or more treatments selected from autologous stem cell transplantation (ASCT); immunomodulators and proteasome inhibitors, alone or in combination; and anti-CD38 agents, optionally four or more prior treatments. In any embodiment, at the time of or prior to administration of the engineered T cell composition, the subject is receiving all three of the following therapies: autologous stem cell transplantation (ASCT), a regimen comprising immunomodulators and proteasome inhibitors, and an anti-CD38 agent.

[0039] In some of the embodiments, at the time of or prior to administration of the composition containing the manipulated T cells, the subject has received all three of the following anti-myeloma treatment regimens: autologous stem cell transplantation (ASCT); immunomodulatory agents and / or proteasome inhibitors, alone or in combination; and anti-CD38 agents. In some of the embodiments, for the purpose of determining the number of prior anti-myeloma treatment regimens, induction with or without bone marrow transplantation and induction with or without maintenance therapy are considered one regimen.

[0040] In some of the embodiments, at the time of or prior to administration of the composition containing the manipulated T cells, the subject is refractory to the last anti-myeloma treatment regimen. In some of the embodiments, refractory myeloma is defined as a confirmed progressive disease during treatment with the last anti-myeloma treatment regimen, or within 12 months from the last dose, as measured after completion of such treatment. In some of the embodiments, refractory myeloma is defined as a confirmed progressive disease during treatment with the last anti-myeloma treatment regimen, or within 60 days from the last dose, as measured after completion of such treatment.

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

[0042] In some of the available embodiments, the subject has received at least one complete cycle of treatment with an anti-myeloma treatment regimen including an immunomodulator and / or proteasome inhibitor, unless progression was the best response to the myeloma treatment regimen. In some of the available embodiments, the subject has received at least two consecutive cycles of treatment with an anti-myeloma treatment regimen including an immunomodulator and / or proteasome inhibitor, unless progression was the best response to the myeloma treatment regimen.

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

[0044] In some of the embodiments, at the time of administration of the composition containing the manipulated T cells, the subject has neither active plasma cell leukemia (PCL) nor a history of plasma cell leukemia (PCL). In some of the embodiments, at the time of administration, the subject has relapsed or is refractory after at least three or at least four prior anti-myeloma treatment regimens. In some of the embodiments, at the time of administration, the subject has been diagnosed with multiple myeloma for approximately four years, or 2–15 years, or 2–12 years. In some of the embodiments, at the time of administration, the subject has received approximately 10, or 3–15, or 4–15 prior anti-myeloma treatment regimens. In some of the embodiments, at the time of administration, the subject is refractory or unresponsive to bortezomib, carfilzomib, lenalidomide, pomalidomide, and / or anti-CD38 monoclonal antibodies. In some of the embodiments, at the time of administration, the subject has received prior autologous stem cell transplantation. In some of the embodiments, at the time of administration, the subject has not received prior autologous stem cell transplantation (ASCT) due to being ineligible for ASCT, or optionally ineligible for age or other confirmed reasons. In some of the embodiments, at the time of administration, the subject has IMWG high-risk cytogenetics. In some of the embodiments, the subject does not have central nervous system infiltration of MM, plasma cell leukemia, Waldenström macroglobulinemia, POEMS (polyneuritis, organomegaly, endocrine abnormalities, monoclonal protein, skin symptoms) syndrome, and / or clinically significant amyloidosis. In some of the embodiments, the subject has not received prior CAR T-cell therapy or prior genetically modified T-cell therapy. In some of the embodiments, the subject has not received prior BCMA-targeted therapy such as anti-BCMA monoclonal antibodies or bispecific antibodies.

[0045] In some of the arbitrary embodiments, the method further comprises the step of obtaining a leukocyte apheresis sample from a subject in order to produce a composition comprising engineered T cells.

[0046] In some of the applicable embodiments, subjects have not received therapeutic doses of corticosteroids, optionally within 14 days or approximately 14 days prior to the time of leukocyte apheresis. In some of the applicable embodiments, subjects have not received immunosuppressive therapy within 4 weeks of leukocyte apheresis, and optionally, immunosuppressive therapy includes calcineurin inhibitors, methotrexate or other chemotherapeutic agents, mycophenolate, rapamycin, immunosuppressive antibodies, e.g., anti-TNF, anti-IL6, or anti-IL6R. In some of the applicable embodiments, subjects have not received autologous stem cell transplantation within 6 months or approximately 6 months prior to the time of leukocyte apheresis.

[0047] In some of the embodiments, the subjects have not achieved complete remission (CR) to prior treatment. In some of the embodiments, the subjects have not achieved an objective response (partial response (PR) or better) to prior treatment. In some of the embodiments, the subjects are identified as having an Eastern Cooperative Oncology Group Performance Status (ECOG PS) of 0 or 1.

[0048] In some of the arbitrary embodiments, the CAR is a variable heavy chain (V) containing heavy chain complementarity determination region 1 (CDR-H1), heavy chain complementarity determination region 2 (CDR-H2), and heavy chain complementarity determination region 3 (CDR-H3) within the sequence shown in SEQ ID NO:116. H ), as well as variable light chain (V) containing light chain complementarity determination region 1 (CDR-L1), light chain complementarity determination region 2 (CDR-L2), and light chain complementarity determination region 3 (CDR-L3) within the sequence shown in SEQ ID NO:119. L ); containing the CDR-H1, CDR-H2, and CDR-H3 sequences shown in SEQ ID NO: 97, 101, and 103 respectively. H , and V containing the CDR-L1, CDR-L2, and CDR-L3 sequences shown in SEQ ID NO: 105, 107, and 108, respectively. L ;V containing the CDR-H1, CDR-H2, and CDR-H3 sequences shown in SEQ ID NO: 96, 100, and 103 respectively.H , and V containing the CDR-L1, CDR-L2, and CDR-L3 sequences shown in SEQ ID NO: 105, 107, and 108, respectively. L ;V containing the CDR-H1, CDR-H2, and CDR-H3 sequences shown in SEQ ID NO: 95, 99, and 103 respectively. H , and V containing the CDR-L1, CDR-L2, and CDR-L3 sequences shown in SEQ ID NO: 105, 107, and 108, respectively. L ;V containing the CDR-H1, CDR-H2, and CDR-H3 sequences shown in SEQ ID NO: 94, 98, and 102 respectively. H , and V containing the CDR-L1, CDR-L2, and CDR-L3 sequences shown in SEQ ID NO: 104, 106, and 108, respectively. L ; or V containing the amino acid sequence of SEQ ID NO:116 H , and V containing the amino acid sequence of SEQ ID NO:119 L ; containing extracellular antigen-binding domain; IgG4 / 2 chimeric hinge or modified IgG4 hinge, IgG2 / 4 chimeric C H 2 regions, and IgG4 C H The structure may include a spacer comprising three regions, optionally approximately 228 amino acids long, optionally the spacer shown in SEQ ID NO:174; a transmembrane domain, optionally a transmembrane domain derived from human CD28; and an intracellular signaling region comprising a cytoplasmic signaling domain of a CD3-zeta (CD3ζ) chain and a costimulatory signaling region containing an intracellular signaling domain or signaling portion of a T cell costimulatory molecule. In some embodiments, the spacer is the spacer shown in SEQ ID NO:174.

[0049] In some of the arbitrary aspects, V H This is either the amino acid sequence of SEQ ID NO:116, or may contain the amino acid sequence of SEQ ID NO:116, V Lis the amino acid sequence of SEQ ID NO:119, or contains the amino acid sequence of SEQ ID NO:119. In some of the any embodiments, the extracellular antigen-binding domain may include scFv. In some of the any embodiments, V H and V L and are linked by a flexible linker. In some of the arbitrary embodiments, scFv is an amino acid sequence It may include a linker containing TIFF2026113544000002.tif4128. In some of any embodiments, V H is V L It is located at the carboxyl terminus. In some of the arbitrary embodiments, scFv is in the amino acid sequence Includes a linker containing TIFF2026113544000003.tif4128.

[0050] In some of the various 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.

[0051] In some of the various embodiments, the nucleic acid encoding the extracellular antigen-binding domain may include the nucleotide sequence of SEQ ID NO:113, the nucleotide sequence having at least 90% sequence identity thereto, or a degenerate sequence of either of the above. In some of the various embodiments, the nucleic acid encoding the extracellular antigen-binding domain may include the nucleotide sequence of SEQ ID NO:115. In some of the various embodiments, V H is V L It is located on the amino end side.

[0052] In some of the various embodiments, the cytoplasmic signaling domain may be the sequence shown at SEQ ID NO:143 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:143, or may include the sequence shown at SEQ ID NO:143 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:143. In some of the various embodiments, the co-stimulatory signaling region may include the intracellular signaling domain or signaling portion of CD28, 4-1BB, or ICOS. In some of the various embodiments, the co-stimulatory signaling region may include the intracellular signaling domain of 4-1BB, optionally human 4-1BB. In some of the various embodiments, the co-stimulatory signaling region may be the sequence shown at SEQ ID NO:4 or an amino acid sequence having at least 90% sequence identity to the sequence shown at SEQ ID NO:4, or may include the sequence shown at SEQ ID NO:4 or an amino acid sequence having at least 90% sequence identity to the sequence shown at SEQ ID NO:4. In some of the various embodiments, the co-stimulatory signaling region is located between the transmembrane domain and the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain.

[0053] In some of the various embodiments, the transmembrane domain is or may include a transmembrane domain derived from human CD28. In some of the various embodiments, the transmembrane domain is or may include the sequence shown at SEQ ID NO:138 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:138, or the sequence shown at SEQ ID NO:138 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:138.

[0054] In some of the embodiments, the CAR comprises, in order from its N-terminus to its C-terminus, an extracellular antigen-binding domain, a spacer, a transmembrane domain, and an intracellular signaling domain. In some of the embodiments, the CAR comprises a variable heavy chain (V) containing heavy chain complementarity-determining region 1 (CDR-H1), heavy chain complementarity-determining region 2 (CDR-H2), and heavy chain complementarity-determining region 3 (CDR-H3) within the sequence shown in SEQ ID NO:116. H ), as well as variable light chain (V) containing light chain complementarity determination region 1 (CDR-L1), light chain complementarity determination region 2 (CDR-L2), and light chain complementarity determination region 3 (CDR-L3) within the sequence shown in SEQ ID NO:119. L ) containing extracellular antigen-binding domain; modified IgG4 hinge, IgG2 / 4 chimeric C H 2 regions, and IgG4 C H It may include a spacer with three regions and a length of approximately 228 amino acids; a transmembrane domain derived from human CD28; and an intracellular signaling region that includes a cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain and a 4-1BB intracellular signaling domain.

[0055] In some of the various embodiments, the CAR may include an extracellular antigen-binding domain comprising an amino acid sequence having at least 90% sequence identity to the sequence shown in SEQ ID NO:114 or the amino acid sequence of SEQ ID NO:114; a spacer comprising an amino acid sequence having at least 90% sequence identity to the sequence shown in SEQ ID NO:174 or the amino acid sequence of SEQ ID NO:174; a transmembrane domain comprising an amino acid sequence having at least 90% sequence identity to the sequence shown in SEQ ID NO:138 or the amino acid sequence of SEQ ID NO:138; and an intracellular signaling region comprising a cytoplasmic signaling region comprising a cytoplasmic signaling region comprising an amino acid sequence having at least 90% sequence identity to the sequence shown in SEQ ID NO:143 or the amino acid sequence of SEQ ID NO:143, and a co-stimulatory signaling region comprising an amino acid sequence having at least 90% sequence identity to the sequence shown in SEQ ID NO:4 or the amino acid sequence of SEQ ID NO:4.

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

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

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

[0059] In some of the various embodiments, prior to administration, the subject was given 20-40 or approximately 20-40 mg / m². 2 The target body surface area can be any 30 or approximately 30 mg / m². 2 Daily administration of fludarabine for 2-4 days, and / or 200-400 mg / m² or approximately 200-400 mg / m² 2 The target body surface area is arbitrarily 300 or approximately 300 mg / m². 2 The patient is receiving lymphocyte depletion therapy, including daily administration of cyclophosphamide for 2-4 days. In some of the available forms, prior to administration, the patient has been given 20-40 or approximately 20-40 mg / m². 2 The target body surface area can be any 30 or approximately 30 mg / m². 2 The patient is receiving lymphocyte depletion therapy, which includes daily administration of fludarabine for 2 to 4 days. In some of the available forms, prior to administration, the subject has been given 200 to 400 or approximately 200 to 400 mg / m². 2 The target body surface area is arbitrarily 300 or approximately 300 mg / m². 2 The subjects are receiving lymphocyte depletion therapy, which includes daily administration of cyclophosphamide over 2 to 4 days. In some of the available embodiments, subjects receive 30 or approximately 30 mg / m² over 3 days, respectively. 2Daily administration of fludarabine based on the target body surface area, and 300 or approximately 300 mg / m² 2 The patient is undergoing lymphocyte depletion therapy, which includes daily administration of cyclophosphamide to the target body surface area.

[0060] In some of the arbitrary embodiments, the method can achieve a specified response or outcome at a specified time after the start of administration, optionally in at least one subject in a cohort of subjects having MM, 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%, the response being selected from the group consisting of objective response (OR), complete response (CR), severe complete response (sCR), very good partial response (VGPR), partial response (PR), and minimal response (MR), and the response or outcome is OR or includes OR, and / or the response or outcome is CR or includes CR.

[0061] In some of the embodiments, the cohort of subjects has at least the same number of prior treatments, prognosis or prognostic factors, subtype, secondary involvement, or one or more other designated patient characteristics as the subjects treated by this method. In some of the embodiments, the response or outcome is sustained for more than 3, 6, 9, or 12 months, or approximately more than 3, 6, 9, or 12 months. In some of the embodiments, the response or outcome determined 3, 6, 9, or 12 months after a designated time point, or approximately 3, 6, 9, or 12 months after a designated time point, is equal to or improved compared to the response or outcome determined at the designated time point.

[0062] In some of the any embodiments, the response or outcome is the absence of neurotoxicity, the absence of cytokine release syndrome (CRS), and / or the absence of macrophage activation syndrome / hemophagocytic lymphohistiocytosis (MAS / HLH), or includes or further includes 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 the any embodiments, the method does not result in the specified toxicity outcome at any specified time after the start of administration in at least one subject, or 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 MM.

[0063] In some of the embodiments, the specified toxicity outcome is neurotoxicity, cytokine release syndrome (CRS), and / or macrophage activation syndrome / hemophagocytic lymphohistiocytosis (MAS / HLH). In some of the embodiments, the specified toxicity outcome is neurotoxicity, and at least 60%, 70%, or 80% of subjects in the cohort of subjects with MM do not develop neurotoxicity. In some of the embodiments, the specified toxicity outcome is grade 3 or higher or grade 4 or higher neurotoxicity. In some of the embodiments, the specified toxicity outcome is grade 3 or higher neurotoxicity, and at least 80%, 85%, 90%, or 95% of subjects in the cohort of subjects with MM do not develop grade 3 or higher neurotoxicity. In some of the 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 the embodiments, at least 15%, 20%, 25%, or 30% of subjects in the cohort of subjects with MM do not develop CRS. In some of the various embodiments, the specified time is 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, or 15 days after the start of administration, or approximately 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, or 15 days, or within 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, or 15 days. In some of the various embodiments, the specified time is 1 month, 3 months, 6 months, 9 months, or 12 months after the start of administration, or approximately 1 month, 3 months, 6 months, 9 months, or 12 months after.

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

[0065] In some of the various embodiments, administration of the composition is optionally performed on an outpatient basis until the subject exhibits or exhibits persistent fever or fever that does not decrease by more than 1°C after treatment with an antipyretic. In some of the various embodiments, administration of the composition is optionally performed without hospitalization and / or without the subject spending the night in a hospital, unless the subject exhibits or exhibits persistent fever or fever that does not decrease by more than 1°C after treatment with an antipyretic. In some of the various embodiments, administration of the composition is optionally performed without hospitalization or spending the night in a hospital, unless the subject exhibits or exhibits persistent fever or fever that does not decrease by more than 1°C after treatment with an antipyretic.

[0066] In some of the embodiments, the composition containing the manipulated T cells is administered parenterally, and optionally intravenously. In some of the embodiments, the subjects are human subjects.

[0067] In some of the arbitrary embodiments, a composition comprising engineered T cells is produced by a manufacturing process comprising: (i) exposing an input composition comprising primary T cells to an irritant reagent comprising an oligomeric particle reagent comprising a plurality of streptavidin mutein molecules under conditions for stimulating T cells, thereby generating a stimulated population, wherein the oligomeric particle reagent comprises a first activator comprising an anti-CD3 antibody or an antigen-binding fragment thereof and a second activator comprising an anti-CD28 antibody or an antigen-binding fragment thereof; (ii) introducing heterologous polynucleotides encoding BCMA-targeting CARs into the T cells of the stimulated population, thereby generating a population of transformed cells; (iii) incubating the population of transformed cells for up to 96 hours; and (iv) collecting T cells from the population of transformed cells, thereby producing a composition of engineered cells, wherein the collection is performed at a time between 24 and 120 hours, including values ​​at both ends, after the commencement of exposure to the irritant reagent. In some embodiments, the input composition comprises autologous T cells enriched by immunoaffinity-based selection of CD3 T cells or CD4 and CD8 T cells from, for example, a blood sample or apheresis (e.g., leukocyte apheresis) sample of interest.

[0068] In some of the embodiments, the composition may include engineered T cells produced by a manufacturing process comprising the step of exposing an input composition containing primary T cells to an irritant reagent comprising an oligomeric particle reagent comprising a plurality of avidin, streptavidin, avidin mutein, or streptavidin mutein molecules under conditions for stimulating T cells, thereby generating a stimulated population, wherein the irritant reagent has the ability to activate 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 co-stimulatory molecules. In some of the embodiments, the manufacturing process further comprises the step of introducing heterologous polynucleotides encoding BCMA-targeting CARs into the T cells of the stimulated population, thereby generating a population of transformed cells. In some of the embodiments, the manufacturing process further comprises the step of incubating the population of transformed cells for up to 96 hours. In some of the embodiments, the incubation step is performed in a basal medium lacking one or more recombinant cytokines.

[0069] In some of the embodiments, the oligomeric particle reagent comprises a first activator comprising an anti-CD3 antibody or an antigen-binding fragment thereof and a second activator comprising an anti-CD28 antibody or an antigen-binding fragment thereof. In some of the embodiments, the anti-CD3 antibody or antigen-binding fragment is Fab, and the anti-CD28 antibody or antigen-binding fragment is Fab. In some of the embodiments, the first activator and the second activator each comprise a streptavidin-binding peptide that reversibly binds the first activator and the second activator to the oligomeric particle reagent, and optionally, the streptavidin-binding peptide comprises an amino acid sequence shown in any one of SEQ ID NO: 266-270. In some of the arbitrary embodiments, the streptavidin mutein molecule is a tetramer of streptavidin mutein containing the amino acid residues Val44-Thr45-Ala46-Arg47 or Ile44-Gly45-Ala46-Arg47, and optionally, the streptavidin mutein contains the sequence shown in any one of SEQ ID NO: 257, 272, 275, 277, 279, 273, or 276. In some of the arbitrary embodiments, the oligomeric particle reagent contains 1,000 to 5,000 streptavidin mutein tetramers, including values ​​at both ends. In some of the arbitrary embodiments, the method further includes the step of adding biotin or a biotin analog prior to cell collection, after incubation, or during incubation.

[0070] In some of the arbitrary embodiments, the manufacturing process further includes the step of collecting T cells from a transformed population and thereby preparing a composition of engineered cells. In some of the arbitrary embodiments, collection is performed at a time point of 24 to 120 hours, including both values, after the start of exposure to the irritant reagent. In some of the arbitrary embodiments, collection is performed at a time point of 48 to 120 hours, including both values, after the start of exposure to the irritant reagent. In some of the arbitrary embodiments, collection is performed when the integrated vector is detected in the genome, but before the copy number of the integrated vector per diploid genome (iVCN) stabilizes. In some of the arbitrary embodiments, collection is performed before the total number of viable cells at collection time is more than three times or approximately three times the total number of viable cells in the stimulated population. In some of the arbitrary embodiments, collection is performed when the total number of viable cells at collection time is three times or approximately three times, two times or approximately two times, or equal to or approximately equal to the total number of viable cells in the stimulated population. In some of the arbitrary embodiments, collection is performed when CD27 + CCR7 + The percentage of cells represents the total number of T cells in the population and the total number of CD3 cells in the population. + T cells, all CD4 cells in the population + T cells, or all CD8 cells in a population + The collection is performed when more than 50% or approximately more than 50% of the T cells or their CAR-expressing cells are present. In some of the arbitrary embodiments, collection is performed on CD45RA + CCR7 + Cells and CD45RA - CCR7 + The percentage of cells represents the total number of T cells in the population and the total number of CD3 cells in the population. + T cells, all CD4 cells in the population + T cells, or all CD8 cells in a population + This is performed when there are more than 60% or approximately more than 60% of T cells or their CAR-expressing cells.

[0071] In some of the arbitrary embodiments, the cells in the administered composition are produced by a manufacturing process to produce an output composition exhibiting predetermined characteristics, and if the iteration of the manufacturing process is performed among several different individual subjects, then optionally multiple output compositions are produced from a human biological sample, and the predetermined characteristics of the output composition among the multiple output compositions are selected from the following characteristics: the average percentage of cells with a memory phenotype in the multiple output compositions is approximately 40% to approximately 65%, approximately 40% to approximately 45%, approximately 45% to The percentages are approximately 50%, 50%–55%, 55%–60%, or 60%–65%; the average percentage of cells with a central memory phenotype in multiple output compositions is approximately 40%–65%, 40%–45%, 45%–50%, 50%–55%, 55%–60%, or 60%–65%; and the percentages in multiple output compositions are CD27+, CD28+, CCR7+, CD45RA-, CD45RO+, CD62L+, CD3+, CD95+, granzyme B-, and / or CD127+. The average percentage of cells is approximately 40%–65%, 40%–45%, 45%–50%, 50%–55%, 55%–60%, or 60%–65%; the average percentage of cells that are CCR7+ / CD45RA- or CCR7+ / CD45RO+ in multiple output compositions is approximately 40%–65%, 40%–45%, 45%–50%, 50%–55%, 55%–60%, or 60%–65%; manipulated CD4+ T cells in multiple output compositions, optionally CAR+ CD In 4+ T cells, the mean percentage of central memory CD4+ T cells is approximately 40%–65%, 40%–45%, 45%–50%, 50%–55%, 55%–60%, or 60%–65%; in manipulated CD8+ T cells of multiple output compositions, and optionally in CAR+CD8+ T cells, the mean percentage of central memory CD8+ T cells is approximately 40%–65%, 40%–45%, 45%–50%, 50%–55%, 55%–60%, or 60%–65%;The mean percentages of central memory T cells, optionally CD4+ central memory T cells and CD8+ central memory T cells in manipulated T cells of and / or multiple output compositions, optionally CAR+ T cells, are approximately 40%–65%, 40%–45%, 45%–50%, 50%–55%, 55%–60%, or 60%–65%.

[0072] In some of the arbitrary embodiments, the administered composition is prepared by a manufacturing process for producing an output composition exhibiting predetermined characteristics, optionally a threshold number of cells expressing CAR in the output composition, in at least about 80%, about 90%, about 95%, about 97%, about 99%, about 100%, or 100% of a human biological sample performed among several different individual subjects. In some of the arbitrary embodiments, the composition containing genetically modified cells does not contain residual beads from the manufacturing process.

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

[0074] This specification also provides a manufactured article comprising a composition containing genetically modified cells expressing a BCMA-targeting chimeric antigen receptor (CAR), and instructions for administering the cell composition according to one of the methods provided herein. [Brief explanation of the drawing]

[0075] [Figure 1]Figure 1 illustrates an exemplary quantification of cellular purity of T cell compositions produced from a non-extended proliferation manipulation process using different donor types (reference, patient), as determined by flow cytometry. Cells were manipulated to express anti-BCMA CAR (BCMA) or mock-transfected (mock). The percentage of CD3+ cells among live CD45+ cells (left), the percentage of NK cells among live CD45+ cells (center), and the percentage of CD19+ cells among live CD45+ cells (right) were determined. [Figure 2] Figures 2 and 3 illustrate the quantitative analysis of cellular phenotypes determined by flow cytometry for expanded and non-expanded growth processes using different donor types (reference, patient). Cells were either manipulated to express anti-BCMA CAR (BCMA) or mock-transfected (mock). Figure 2 shows the percentage of CD3+CD8+ and CD3+CD4+ cells among live CD45+ cells (left side), and the percentage of CD8+CAR+ and CD4+CAR+ cells among live CD45+ cells (right side). [Figure 3] Figure 3 shows the ratio of CD4+ cells to CD8+ cells, and the ratio of CD4+CAR+ cells to CD8+CAR+ cells. [Figure 4] Figure 4 shows an exemplary quantification of cell viability of T cell compositions produced from a non-extended proliferation manipulation process using different donor types (reference, patient), as determined by flow cytometry. Cells were manipulated to express anti-BCMA CAR (BCMA) or mock-transfected (mock). The percentage of aCas3+ cells among CD3+ cells was determined. [Figure 5-1]Figure 5A illustrates the exemplary relationship between the copy number per cell in the total cells, as assessed by a standard VCN (with PFGE) and an iVCN (without PFGE), in cell compositions produced using either an expansion process (○) or a non-expansion process (●) from primary T cells derived from different human donors engineered to express CAR. Figures 5B-5C show the relationship between the copy number per cell in the cell compositions, as assessed by a standard VCN (Figure 5B) or an iVCN (Figure 5C), and the surface expression of CAR, indicated by the percentage of CAR-expressing CD3+ cells (%CD3+CAR+) in living CD45+ cells, as assessed by flow cytometry. [Figure 5-2] See the explanation in Figure 5-1. [Figure 6A] Figures 6A–6B show exemplary percentages of cellular phenotypes obtained as a result of expanded and non-expanded growth manipulation processes using different donor types (reference, patient). Cells were manipulated to express anti-BCMA CAR (BCMA) or mock-transfected (mock). Figure 6A shows exemplary percentages of CD45RA+CCR7+ cells (top left) among aCas-CD8+CAR+ cells and aCas-CD4+CAR+ cells, CD45RA-CCR7+ cells (top right) among aCas-CD8+CAR+ cells and aCas-CD4+CAR+ cells, CD45RA-CCR7- cells (bottom left) among aCas-CD8+CAR+ cells and aCas-CD4+CAR+ cells, and CD45RA+CCR7- cells (bottom right) among aCas-CD8+CAR+ cells and aCas-CD4+CAR+ cells. Figure 6B shows the exemplary percentage of CD27+CCR7+ cells among aCas-CD8+CAR+ cells and aCas-CD4+CAR+ cells. [Figure 6B] See the explanation in Figure 6A. [Figure 7]Figure 7 shows exemplary proportions of the T cell memory phenotype, defined by the surface expression of CD45RA and CCR7, in CAR T cells derived from donor-matched non-extended growth process products and extended growth process products. CAR T cells were produced from CD4+ and CD8+ T cells derived from one healthy donor (HD1) or three multiple myeloma patients (MM1, MM2, or MM3). [Figure 8A] Figures 8A–8C illustrate the in vitro proliferative capacity of cells produced by the non-expanding and expanding proliferation processes after prolonged anti-BCMA CAR-dependent stimulation with agonist antibodies. CAR T cells were produced from CD4+ and CD8+ T cells derived from one healthy donor (HD1) or three multiple myeloma patients (MM1, MM2, MM3), and the number of viable cells was determined every two days for 10 days after stimulation with agonist antibody-coated microbeads. Figure 8A shows the expansion factor change calculated by dividing the daily count by the number of starting cells. Figure 8B shows the CAR T cell count converted to AUC for comparison between products (non-expanding process products, expanding process products, mock) or between donors. Statistical significance was determined by the Mann-Whitney U test; *p<0.05. Figure 8C shows the expansion factor for each donor, calculated by dividing the daily expansion factor in the non-expanding process product group by the donor-matched expanding process product value. [Figure 8B] See the explanation in Figure 8A. [Figure 8C] See the explanation in Figure 8A. [Figure 9A]Figures 9A–9E illustrate the quantitative determination of intracellular IL-2, IFNγ, or TNF cytokine production from anti-BCMA CAR T cells derived from donor-matched non-extended growth process products and extended growth process products, as measured by flow cytometry (Figures 9A–9D) and secreted cytokines (Figure 9E). CAR T cells derived from one multiple myeloma patient or two healthy donors after 5 hours of culture with agonist antibodies. Figures 9A–9C represent the frequency of CAR-positive cells expressing a single cytokine (Figures 9A–9C) or Boolean logic-gated triple-positive cells (Figure 9D) within the CD4+CAR+ or CD8+CAR+ population. Cytokine protein secretion was measured by multiplex immunoassay quantification of secreted cytokine concentrations after 24 hours of culture with MM.1S BCMA-positive target cells (Figure 9E, showing total protein secretion measured in the culture supernatant). Statistical significance was assessed by Mann-Whitney. *p<0.05. [Figure 9B] See the explanation in Figure 9A. [Figure 9C] See the explanation in Figure 9A. [Figure 9D] See the explanation in Figure 9A. [Figure 9E] See the explanation in Figure 9A. [Figure 10] Figure 10A shows the exemplary cytolytic activity of anti-BCMA CAR T cells engineered by a non-extended proliferation process or an extended proliferation process at different effector-to-target ratios. Area under the curve (AUC) was compared for each individual arm (left side of Figure 10B) or by the manufacturing process (right side of Figure 10B) (statistical significance by Mann-Whitney test; *p<0.05). [Figure 11A]Figures 11A–11B show exemplary tumor burden and circulating CAR-T cells over time in an OPM-2 myeloma model after treatment with anti-BCMA CAR-T cell compositions generated by matching donor non-extended growth and extended growth processes. Tumor growth from day 1 (pre-treatment) to approximately day 53 post-treatment is shown in BLI (photons / second; y-axis) (Figure 11A) or calculated from the area under the BLI curve (AUC) for each group (Figure 11B). Differences were compared using the Mann-Whitney U test; *p<0.05. [Figure 11B] See the explanation in Figure 11A. [Figure 12A] Figures 12A–12B show exemplary CAR T cell dynamics and circulating CAR-T cell counts over time in an OPM-2 myeloma model after treatment with anti-BCMA CAR-T cell compositions generated by matched donor non-extended growth and extended growth processes. Figure 12A shows the circulating anti-BCMA CAR-T cell count per μl of blood after treatment for each group. Figure 12B shows the circulating anti-BCMA CAR-T cell count per μl of blood at the indicated time point after treatment for each group (non-extended growth, NE; extended growth, E). Differences were compared using the Mann-Whitney U test; *p<0.05. [Figure 12B] See the explanation in Figure 12A. [Modes for carrying out the invention]

[0076] Detailed explanation This specification provides methods and uses of engineered cells expressing anti-BCMA recombinant receptors (e.g., CARs), as well as pharmaceutically active compositions and formulations thereof, in the treatment of diseases, conditions, and disorders in which BCMA is expressed, most specifically in hematological malignancies, such as multiple myeloma (MM). In aspects of the methods provided, a therapeutic T-cell composition containing engineered cells is administered to a subject having MM, for example, by adoptive cell therapy, such as adoptive T-cell therapy. In certain aspects of the methods provided, the T cells are engineered with CARs that target BCMA. In several aspects, the methods and uses provide or achieve, compared to certain alternative methods, improved response and / or more sustained response or efficacy and / or reduced risk of toxicity or other adverse effects, for example, in a particular group of subjects being treated. In some embodiments, the method is advantageous due to the administration of a specified number or relative number of manipulated cells, the administration of a cell composition having predetermined characteristics (e.g., a ratio of specific cell types), the administration of a high specific percentage of poorly differentiated cells (e.g., naive-like or central memory cells or cells in an early differentiated state, e.g., CCR7+CD27+ cells), and the treatment of specific patient populations, such as those having a specific risk profile, disease stage, and / or prior treatment history, and / or combinations thereof.

[0077] In some embodiments, the method and use include administering to a subject T cells expressing a genetically modified (recombinant) cell surface receptor, typically a chimeric receptor such as a chimeric antigen receptor (CAR), which is expressed by, associated with, and / or specific to MM and / or the cell type from which it is derived, BCMA expressed by MM and / or the cell type from which it is derived. The cells are generally administered as a composition formulated for administration, and the method generally involves administering the cells to the subject once or multiple times, with each dose containing a specific number or relative number of cells or modified cells. Optionally, the BCMA-targeted CAR+ modified cells in the composition include two or more subtypes, e.g., CD4 T cells and CD8 T cells, in a predetermined ratio or composition. In a particular embodiment, the cell composition for use or administration in the provided method comprises primary T cells engineered to express a BCMA-targeted CAR, comprising (i) a low percentage (e.g., less than 40%, less than 30%, less than 20%, or less than 10%) of exhausted cells and / or cells exhibiting exhaustion-related markers or phenotypes, and / or (ii) 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, e.g., naive-like T cells, central memory T cells, or long-lived memory T cells. In the provided embodiment, the features of the composition and the provided method result in improved or enhanced survival, growth, persistence, and / or antitumor activity compared to other BCMA-targeted CAR T cell therapy administrations with a high percentage of exhausted cells and / or a high number of cells exhibiting exhaustion-related phenotypes and / or a low percentage of specific T cells such as naive-like T cells, central memory T cells, or long-lived memory T cells.In the provided embodiment, the features of the composition and the provided method result in improved therapeutic efficacy, such as an increased percentage of patients achieving complete response (CR), compared to methods involving the administration of other BCMA-targeted CAR T-cell therapies that have a high percentage of exhausted cells and / or a high number of cells exhibiting exhaustion-related phenotypes and / or a low percentage of specific T cells such as naive-like T cells, central memory T cells, or long-lived memory T cells. In the provided method, the features of the composition and the provided method result in improved clinical persistence of therapeutic responses, such as CR, such as responses that persist for a period of time after the start of treatment, compared to methods involving the administration of other BCMA-targeted CAR T-cell therapies that have a high percentage of exhausted cells and / or a high number of cells exhibiting exhaustion-related phenotypes and / or a low percentage of memory-like T cells such as naive-like T cells, central memory T cells, or long-lived memory T cells. In certain embodiments, the use or administration of the BCMA-targeted CAR T cell composition provided in the provided method can be achieved with a dose of cells less than half, for example, one-fifth or one-tenth, of a dose of a reference BCMA-targeted CAR T cell composition (e.g., one manipulated with the same or a similar CAR, e.g., having the same antigen-binding domain) (wherein the reference BCMA-targeted CAR T cell composition has a high percentage of exhausted cells and / or a high number of cells exhibiting exhaustion-related phenotypes and / or a low percentage of memory-like T cells such as naive-like T cells, central memory T cells, or long-lived memory T cells). In some embodiments, the reference BCMA-targeted CAR T cell composition is a composition produced ex vivo by a process that involves a step of culturing the cells during the process of producing the cells under conditions for expanded proliferation, e.g., cell proliferation or doubling of the cell population (e.g., doubling of the cells in the population 2, 3, 4, 5, 6, 7, 8, 9, 10 or more times compared to the start of the process).

[0078] In some embodiments, the BCMA-directed CAR T cell compositions for use in the methods and uses provided are produced by a relatively short process that does not involve the step of culturing cells under conditions designed for cell expansion or proliferation. Different processes can be utilized to produce compositions containing genetically engineered T cell populations, including the generation of engineered T cells expressing CARs, and these typically involve a step designed to culture cells for cell expansion or increased cell proliferation, or for that purpose. However, in certain aspects, some of these processes may require long or relatively long periods of time to generate the engineered cells. In addition, in various aspects, some existing processes may have varying time required to successfully produce engineered T cells suitable for cell therapy, which makes it difficult to coordinate the administration of cell therapy. In certain aspects, some of these processes may produce cell populations with a relatively high percentage or a relatively large amount of exhausted cells, differentiated cells, or low-titer cells. The BCMA-directed CAR T cell compositions provided for use in the methods provided address one or more of these challenges.

[0079] In certain embodiments, the method provided is used in conjunction with a process for efficiently producing or generating engineered cells suitable for use in cell therapy. In some embodiments, the provided composition containing BCMA-directed CAR engineered T cells is produced by a process that does not require any additional steps for cell expansion and proliferation, for example, without expansion and proliferation unit manipulation and / or without steps intended to cause cell expansion and proliferation. In aspects of the process for producing a BCMA-directed CAR T cell composition, the process includes one or more steps for producing a population of engineered T cells that can be collected or formulated for use as a composition for cell therapy by stimulating and genetically manipulating (e.g., transforming, transducing, or transfecting) T cells. In certain embodiments, the process includes a step of transducing cells using a viral vector (e.g., a lentiviral vector) containing nucleic acids encoding BCMA-directed CARs. In some embodiments, the provided process results in the stable integration of heterologous nucleic acids (expressed from the viral vector) into the cell genome. In several aspects, the provided process generates engineered BCMA-targeted CAR T cells with improved titer compared to engineered T cell compositions produced by alternative processes, such as processes that involve cell expansion and proliferation.

[0080] In certain contexts, the time required for the process to produce the composition to be provided can be measured from the time when the input cell population or the cells of the input composition, such as T cells, are first brought into contact with or exposed to stimulating conditions (as described in Section II-C, for example) (this is referred to herein as the start of the stimulation or stimulating process, and also as exposure to an irritating reagent, for example, when exposure to an irritating reagent is initiated). In some embodiments, the time required to harvest or collect the output population containing the manipulated cells (referred herein as the output composition, or composition of the manipulated cells, such as manipulated T cells) is measured from the start of stimulation. In certain embodiments, the process duration is 120 hours, 108 hours, 96 hours, 84 hours, 72 hours, 60 hours, 48 ​​hours, 36 hours, or 30 hours, or approximately 120 hours, 108 hours, 96 hours, 84 hours, 72 hours, 60 hours, 48 ​​hours, 36 hours, or 30 hours, or less than 120 hours, 108 hours, 96 hours, 84 hours, 72 hours, 60 hours, 48 ​​hours, 36 hours, or 30 hours. In certain embodiments, the process duration is 5 days, 4 days, 3 days, 2 days, or 1 day, or approximately 5 days, 4 days, 3 days, 2 days, or 1 day, or less than 5 days, 4 days, 3 days, 2 days, or 1 day. In certain embodiments, the manipulated cells, e.g., cells in an output composition or output population, are more potent, persistent, or naive-like than cells manipulated in a process requiring a longer duration. In some aspects, the time required for the provided process, for example, the time required to generate or produce the manipulated T cell population, is 2, 3, 4, 5, 6, 7, or 7 days or more shorter than the time required for some existing processes, or by approximately 2, 3, 4, 5, 6, 7, or 7 days or more, or by at least 2, 3, 4, 5, 6, 7, or 7 days or more.In some embodiments, the time required for the provided process is 75%, 60%, 50%, 40%, 30%, 25%, 15%, or 10% of the alternative or existing process, or approximately 75%, 60%, 50%, 40%, 30%, 25%, 15%, or 10%, or less than 75%, 60%, 50%, 40%, 30%, 25%, 15%, or 10%.

[0081] In certain embodiments, the provided process is carried out on a population of cells isolated, enriched, or selected from a biological sample, e.g., CD3+, CD4+, and / or CD8+ T cells. In some aspects, the provided method can produce or generate a composition of engineered T cells within a shorter time compared to other methods or processes from the time the biological sample is collected from the subject. In some embodiments, the provided method can produce or generate engineered T cells within 10, 9, 8, 7, 6, 5, 4, 3, or 2 days, or within approximately 10, 9, 8, 7, 6, 5, 4, 3, or 2 days, or within 120, 96, 72, or 48 hours, or within approximately 120, 96, 72, or 48 hours, whenever the biological sample or enriched, isolated, or selected cells are cryopreserved and stored.

[0082] In certain embodiments, a process for producing or manipulating a T cell population includes a cell stimulation step, for example, prior to transduction by a viral vector. In aspects of the provided process, stimulation is carried out using oligomeric stimulant reagents, such as streptavidin mutein oligomers to which stimulant binding agents, e.g., anti-CD3 / anti-CD28, are immobilized or attached. Existing reagents for stimulating T cells in vitro, for example, in the absence of exogenous growth factors or in small amounts of exogenous growth factors, are known (see, e.g., U.S. Patent No. 6,352,694 B1 and European Patent EP 0 700 430 B1). Generally, such reagents may use beads with a diameter greater than 1 μm, e.g., magnetic beads, to which various binding agents (e.g., anti-CD3 antibodies and / or anti-CD28 antibodies) are immobilized. However, in some cases, such magnetic beads are difficult to incorporate into methods for stimulating cells under conditions required for clinical or therapeutic purposes. This is because it is necessary to ensure that the magnetic beads are completely removed before administering the expanded T cells to the target. In some cases, such removal, such as by exposing cells to a magnetic field, can reduce the yield of viable cells available for cell therapy. In certain cases, the incubation time of such reagents, such as irritating reagents containing magnetic beads, with cells must be kept to a minimum so that a sufficient number of T cells can be detached from the irritating reagent.

[0083] Such potential limitations are overcome in the provided process utilizing oligomeric irritants, such as streptavidin mutein polymer. For example, in some embodiments, the provided process avoids or reduces the risk of irritants, such as those containing magnetic beads, remaining in the output cells generated or produced by the process. In some embodiments, this also means that a GMP-compliant process can be established more easily than in other methods, such as those requiring additional measures to ensure that the final manipulated T cell population does not contain beads. In some embodiments, this can be achieved readily by adding a substance that dissociates the oligomeric irritant from cells, such as a competing reagent, for example, by simply rinsing or washing the cells, for example, by centrifugation. Therefore, in some aspects, the removal or separation of oligomeric irritants from cells by adding a substance or competing reagent results in little to no cell loss compared to the removal or separation of irritants based on beads. In some cases, the timing of removal or separation of oligomeric irritants is unconstrained or less constrained than that of bead-based removal or separation. Therefore, in some cases, oligomeric irritants can be removed or separated from cells at any point or stage of the provided process.

[0084] In some aspects, the use of oligomeric stimulants (e.g., anti-CD3 / anti-CD28 streptavidin mutein oligomers) may result in a generally reduced stimulating signal compared to alternative stimulants such as anti-CD3 / anti-CD28 paramagnetic beads. Processes provided with weaker or reduced stimulation may produce potent, sustained, or effective engineered CAR+ T cells that are as potent, sustained, or even more potent than those produced by processes with stronger stimulant conditions or larger or higher concentrations of stimulants, such as those produced after stimulation with anti-CD3 / anti-CD28 paramagnetic beads. In addition, in some embodiments, stimulating cells with smaller or relatively smaller amounts of oligomeric stimulants may increase the titer, potency, or persistence of the resulting engineered cell population compared to processes using larger amounts of oligomeric stimulants. In such embodiments, such effects are expected to persist even at doses low enough to reduce the expression of activation markers or the percentage of cells positive for activation markers during and after the process.

[0085] In certain embodiments, an output composition or population of T cells containing engineered T cells, such as T cells expressing recombinant receptors such as chimeric antigen receptors produced or generated by the provided process, is particularly effective or potent when used as cells for cell therapy. For example, in several aspects, an output composition containing engineered T cells, such as CAR+ T cells, generated by the provided process has a much higher titer and / or proliferative capacity than engineered T cells produced or generated by alternative existing processes. In several aspects, an output composition containing engineered T cells, such as CAR+ T cells, generated by the provided process has higher antitumor activity or anticancer cell activity than engineered T cells, such as CAR+ T cells, produced by alternative or existing methods.

[0086] In certain embodiments, a process for producing a BCMA-targeted T cell composition that does not involve a step of expanding and growing cells to a threshold volume or concentration offers further advantages. In some aspects, a protocol that does not rely on a step of expanding and growing cells to increase the number or concentration of cells from a starting cell population, e.g., an input population, does not require incubation or culture, which can vary between cell populations. For example, in some embodiments, cell populations obtained from subjects with different diseases or disease subtypes, such as patients with MM including high-risk, aggressive, and / or R / R MM, are thought to divide or expand at different rates. In certain aspects, eliminating a potentially indeterminate step that requires cell expansion and growth allows for tight control over the overall process duration. In certain embodiments, variability in process duration is reduced or eliminated, and in some aspects, this can improve the coordination of appointments and procedures among physicians, patients, and technicians to facilitate autologous cell therapy.

[0087] In some embodiments, the methods provided involve the treatment of a specific group or subset of subjects, e.g., subjects identified as having a high-risk disease, e.g., a high-risk hematological malignancy or high-risk MM. In some aspects, the methods treat subjects with a certain form of poor-prognosis MM, e.g., subjects with relapsed or refractory (R / R) MM and / or poor-prognosis MM. In some aspects, the methods treat subjects with relapsed or refractory (R / R) MM. In certain aspects, the manipulated cells are autologous to the subject and are administered after production by an ex vivo process that is a shortened ex vivo process compared to existing methods, does not include or requires a culture step to expand and proliferate the cells during the method of producing the manipulated cells, and / or allows for administration of CAR-manipulated T cell compositions with a lower degree of differentiation and lower doses. As a result, the methods provided are advantageous compared to existing methods. This is because it can shorten the time until manipulated T-cell therapy becomes available to patients, especially those in need of treatment, such as those who have relapsed after one or more other prior treatments for the disease or condition, or those who are refractory to treatment. In some aspects, the methods, compositions, uses, and manufactured articles provided achieve improved or superior responses compared to available therapies. In some aspects, the comparison for the improved or superior response is the current standard of care (SOC).

[0088] Multiple myeloma (MM) is a hematological malignancy characterized by clonal proliferation and accumulation of malignant plasma cells in the bone marrow and the development of osteolytic lesions (Palumbo et al., N Engl J Med. 2011;364(11):1046-60). It accounts for approximately 10% of all hematological malignancies. In the United States (US), approximately 32,110 new cases were diagnosed in 2019, and an estimated 12,960 deaths were attributed to MM (Siegel et al., CA Cancer J Clin. 2019;69(1):7-34).

[0089] The median age at diagnosis is 69 years, and less than 15% of newly diagnosed patients are under 55 years old (SEER Cancer Stat Facts: Myeloma Web site, https: / / seer.cancer.gov / statfacts / html / mulmy.html, accessed March 8, 2019). The clinical features of the symptomatic disease are summarized in the so-called "CRAB criteria," which consist of elevated calcium, renal dysfunction, 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; some patients progress rapidly despite treatment, while others do not require treatment for several years. The median overall survival (OS) is 5–6 years (Nandakumar et al., JCO 2019;37:15_suppl:8039).

[0090] Novel agents (i.e., immunomodulators, proteasome inhibitors, anti-CD38 or SLAMF7-targeted monoclonal antibodies), used alone or in combination with conventional therapies, have led to significant improvements in clinical outcomes for MM patients. However, despite these recent advances, MM remains an incurable disease characterized by relapses and high mortality rates due to the emergence of drug-resistant clones (Cho et al., Front Immunol. 2018;9:1821, Cornell et al., Bone Marrow Transplant 2016;51(4):479-91). Median overall survival (OS) is short for patients with MM that is relapsed and / or refractory (R / R) to available treatments. Therefore, further novel therapeutic approaches are needed to overcome relapses and improve survival outcomes in MM patients.

[0091] B cell maturation antigen (BCMA), a member of the tumor necrosis factor (TNF) receptor superfamily, is a cell surface protein expressed on plasma cells and is involved in regulating B cell maturation and differentiation into plasma cells. This is induced during plasma cell differentiation in parallel with the loss of expression of the associated B cell activator receptor (BAFF-R). BCMA binding to its ligands, B cell activator (BAFF) and proliferation-inducing ligand (APRIL), leads to plasma cell survival and enhances humoral immunity.

[0092] BCMA is an attractive therapeutic target because it is highly expressed on MM cell lines and cells from MM patients, and its expression appears to increase with disease progression (Tai et al., Immunotherapy 2015;7(11):1187-99). Importantly, the BCMA protein is not expressed in hematopoietic stem cells, naive 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). Therefore, toxicity associated with on-target / off-tumor interactions is reduced with agents targeting BCMA.

[0093] A major challenge in CAR T cell development is ensuring consistent proliferation, sustained growth, and the production of products that mediate a sustained antitumor response after injection. BCMA-targeted CAR T cells have been evaluated for the treatment of multiple myeloma (MM). In preclinical studies, T cells transduced using BCMA-targeted chimeric antigen receptor (CAR) constructs proliferated and produced high levels of cytokines (e.g., interferon-gamma [IFN-γ], TNFα, interleukin-2 [IL-2]) when stimulated with BCMA-expressing target cells. In addition, BCMA-targeted CAR T cells killed BCMA-expressing MM cells and eradicated BCMA-expressing tumors in a mouse xenograft model (Carpenter et al., Clin Cancer Res. 2013;19(8):2048-60). The survival of CAR-manipulated T cells after administration of BCMA-targeted CAR T cells and the persistence of responses in patients with multiple myeloma remain challenges.

[0094] In certain embodiments, the methods provided herein are based on the administration of 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 CD3 zeta-derived signaling domain and also contains a 4-1BB costimulatory domain.

[0095] The methods provided are based on the finding that the poorly differentiated state of adoptively transferred T cells affects the sustained ability of these cells and can promote sustained antitumor immunity. In some embodiments, the BCMA-targeted CAR+ manipulated T cell compositions provided are produced by a method in which the cells are not cultured under conditions for expansion and proliferation, resulting in a limited or reduced population doubling number of the final manipulated output composition and consequently a less differentiated product. Furthermore, the compositions provided are also produced by a process that results in a stably incorporated vector copy number (iVCN) to ensure consistent and reliable expression of CAR, thereby yielding a consistent cell product for administration to the target and reducing dose variability among CAR-expressing cells. In contrast, most protocols for T cell manipulation typically involve the T cells being expanded and proliferated ex vivo for 9–14 days or longer. The data provided, illustrated herein, support a model in which CAR T cell products with an increased composition of less differentiated memory T cells may exhibit enhanced sustained antitumor activity. These findings demonstrate that strategies aimed at minimizing effector differentiation in CAR T cell products may lead to improved clinical efficacy. This specification provides embodiments that can achieve such objectives.

[0096] The findings herein support treating subjects with high-risk diseases with BCMA-targeted CAR T-cell therapy according to the methods presented. For example, subjects with MM, including high-risk MM patients such as those with relapsed / refractory (R / R) MM, can be treated according to the methods provided. In some embodiments, the methods provided can be used to treat subjects with multiple prior treatment histories (e.g., those who have received one, two, three, four or more prior treatments to treat the disease).

[0097] Any publications referenced herein, including patent documents, scientific articles, and databases, are incorporated herein in whole for all purposes to the same extent as if each individual publication were incorporated herein by reference individually. If any definition provided herein conflicts with or otherwise contradicts any definition provided herein in a patent, patent application, published patent application, or other publication incorporated herein by reference, the definition provided herein shall prevail over the definition incorporated herein by reference.

[0098] Section headings used in this specification serve only structural purposes and should not be construed as limiting the subjects described.

[0099] I. Methods and Use of BCMA-Targeted Cell Therapy in Multiple Myeloma Methods of treatment comprising the step of administering engineered cells or compositions containing engineered cells, for example, engineered T cells. Methods and uses of provided BCMA-targeted CAR-manipulated cells (e.g., T cells) and / or compositions, including methods of treatment for subjects having multiple myeloma (MM), including high-risk MM, for example, R / R MM, also provided, methods and uses involving the administration of engineered cells and / or compositions. In some embodiments, methods and uses of provided BCMA-targeted CAR-manipulated cells (e.g., T cells) and / or compositions, including methods of treatment for subjects having R / R MM that have failed at least two or more previous therapies. In certain embodiments, the method comprises the step of administering a dose of T cells, including CD4+ and CD8+ T cells, to a subject, wherein the T cells contain chimeric antigen receptors (CARs) that specifically bind to BCMA.

[0100] In some embodiments, the method and use involve administering cells expressing a genetically engineered (recombinant) cell surface receptor, typically a chimeric receptor such as a chimeric antigen receptor (CAR), which is expressed by MM and / or a cell type from which it originates, is associated with MM and / or a cell type from which it originates, and / or recognizes BCMA specific to MM and / or a cell type from which it originates. The cells are typically administered in a composition formulated for administration. In some embodiments, the cells are collected from a subject prior to the procedure for the purpose of manipulating the cells with a BCMA-targeted recombinant receptor (e.g., CAR). In some embodiments, the cells are collected by leukocyte apheresis. In some aspects, the cells are manipulated by an ex vivo method (hereinafter also referred to as a non-expanding method) that does not involve a step of culturing the cells for expansion and proliferation. Exemplary non-expanding methods for manipulating the CAR-expressing therapeutic composition provided are described in Section II.C.

[0101] In some embodiments, the disease or condition being treated is high-risk multiple myeloma (MM). In some embodiments, the subject has a measurable disease indicated by: serum M protein levels higher than or equal to 0.5 g / dL as determined by serum protein electrophoresis (SPEP); urinary M protein levels higher than or equal to 200 mg / 24 hours as determined by urinary protein electrophoresis (UPEP); serum free light chain (SFLC) levels higher than or equal to 10 mg / dL accompanied by an abnormal kappa / lambda ratio; or any combination of the above. In some embodiments, the subject prior to leukocyte apheresis has a measurable disease indicated by: serum M protein levels lower than 0.5 g / dL; urinary M protein levels lower than 200 mg / 24 hours; and SFLC levels higher than or equal to 10 mg / dL accompanied by an abnormal kappa / lambda ratio.

[0102] In some embodiments, subjects prior to leukocyte apheresis have 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 index may be used to assess or select subjects for treatment, e.g., subjects who have had poor outcomes from previous 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 functionality in terms of their ability to care for themselves, their daily activity level, and their physical capacity (e.g., walking, working). In some embodiments, an ECOG performance status of 0 indicates that the subject is able to perform normal activities. In some cases, a subject with an ECOG performance status of 1 exhibits some limitations in physical activity, but the subject is fully able to walk. In some cases, patients with an ECOG performance status of 2 are able to walk more than 50% of the time. In some cases, subjects with an ECOG performance status of 2 may also be able to care for themselves; see, for example, Sorensen et al., (1993) Br J Cancer 67(4)773-775. The criteria reflecting ECOG performance status are listed in Table 1 below.

[0103] (Table 1) ECOG Performance Status Criteria TIFF2026113544000004.tif47160

[0104] In some embodiments, prior to the administration of the BCMA-targeted CAR T cell composition provided, for example at that time, the subject has relapsed after remission following treatment with one or more lines of prior therapy for MM, or is refractory to said line. In any embodiment, prior to leukocyte apheresis in connection with the manipulation of the BCMA-targeted CAR T cell composition, the subject has relapsed after remission following treatment with one or more lines of prior therapy for treating MM, or is refractory to said line. Thus, in certain embodiments, prior to the time of treatment, for example prior to leukocyte apheresis, the subject has R / R MM. In some embodiments, the subject has been previously treated with a disease or condition, e.g., MM, or a therapy or therapeutic agent targeting it, prior to the administration of cells expressing recombinant receptors. In some embodiments, the subject has been previously treated with hematopoietic stem cell transplantation (HSCT), e.g., allogeneic HSCT or autologous HSCT. In some embodiments, the subject has a poor prognosis after treatment with standard therapy and / or has failed one or more lines of prior therapy. In some embodiments, the subjects have been treated with or previously received at least one, two, three, four or more, or at least about one, two, three, four or more, or about one, two, three, four or more, other therapies to treat MM, e.g., high-risk MM. In some embodiments, the subjects have been treated with or previously received therapies including CD38-targeted agents (e.g., anti-CD38 antibodies). In some aspects, the subjects have relapsed after an initial response of complete response (CR) or partial response (PR) to prior therapy. In some embodiments, the subjects are refractory to treatment with at least one or more prior therapies, and the refractory treatment is the best response of stable disease (SD) or progressive disease (PD) after prior therapy.

[0105] In some embodiments, the subject has relapsed or is refractory to the line of therapy after remission following treatment with one or more lines of prior therapy for the disease or condition. In some embodiments, the subject is considered refractory to the line of therapy if progressive disease is confirmed during or within 60 days of completing the last dose of the line of therapy. In some embodiments, the subject has progressive disease confirmed during or within 60 days of completing the last dose of the line of therapy. In some embodiments, the subject has progressive disease confirmed during or within 12 months of completing the last dose of the line of therapy. In some embodiments, the subject has progressive disease confirmed during or within 6 months prior to administration of the engineered T-cell composition, and the subject is refractory or unresponsive to the most recent line of therapy for treating MM.

[0106] In some embodiments, the subject has relapsed or is refractory to at least two lines of treatment for the disease or condition after achieving remission. In some embodiments, the subject has relapsed or is refractory to at least three lines of treatment for the disease or condition after achieving remission.

[0107] In some embodiments, subjects have relapsed after remission following treatment with autologous stem cell transplantation (ASCT) or are refractory to ASCT. In some embodiments, subjects have not undergone ASCT prior to leukocytosis due to age or other factors.

[0108] In some embodiments, the subjects have relapsed or are refractory to immunomodulatory agents after remission following at least one cycle of treatment with an immunomodulator. Examples of immunomodulatory agents include, but are not limited to, thalidomide, lenalidomide, and pomalidomide. In some embodiments, the subjects have relapsed or are refractory to immunomodulatory agents after remission following at least two consecutive cycles of treatment with an immunomodulator. In some embodiments, the subjects have relapsed or are refractory to immunomodulatory agents after remission following at least one completed cycle of treatment with an immunomodulator.

[0109] In some embodiments, the immunomodulator is an immune checkpoint inhibitor. In some embodiments, the immunomodulator is an immunomodulatory antibody. Examples of immune checkpoint inhibitors include tremelimumab (CTLA-4 blocking antibody, also known as tisilimubab, 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). Examples of immunomodulatory antibodies include daclizumab (Xenapax), bevacizumab (Avastin®), basiliximab, ipilimumab, nivolumab, pembrolizumab, MPDL3280A, pizilizumab (CT-011), MK-3475, BMS-936559, MPDL3280A (atezolizumab), tremelimumab, IMP321, BMS-986016, LAG525, urerumab, PF-05082566, TRX518, MK-4166, dasetuzumab (SGN-40), lucatumumab (HCD122), and SEA-CD40. Examples include, but are not limited to, CP-870, CP-893, MEDI6469, MEDI6383, MOXR0916, AMP-224, MSB0010718C (avelumab), MEDI4736, PDR001, rHIgM12B7, urocuplumab, BKT140, valirumab (CDX-1127), ARGX-110, MGA271, lirirumab (BMS-986015, IPH2101), IPH2201, ARGX-115, emactuzumab, CC-90002, and MNRP1685A or their antibody-binding fragments.Other exemplary immunomodulatory agents include, for example, aftuzumab (available from Roche®), pegfilgrastim (Neulasta®), lenalidomide (CC-5013, Revlimid®), thalidomide (Thalomid®), actimide (CC4047), and IRX-2 (a mixture of human cytokines including interleukin-1, interleukin-2, and interferon-gamma, CAS 951209-71-5, available from IRX Therapeutics).

[0110] In some embodiments, the subjects have relapsed or are refractory to the disease after remission following at least one cycle of treatment with a proteasome inhibitor. Examples of proteasome inhibitors include, but are not limited to, bortezomib, carfilzomib, and ixazomib. In some embodiments, the subjects have relapsed or are refractory to the disease after remission following at least two consecutive cycles of treatment with a proteasome inhibitor.

[0111] In some embodiments, the subjects have relapsed or are refractory to relapse after remission following at least two consecutive cycles of treatment with an immunomodulator alone and at least two consecutive cycles of treatment with a proteasome inhibitor alone. In some embodiments, the subjects have relapsed or are refractory to relapse after remission following at least two consecutive cycles of treatment with a combination of an immunomodulator and a proteasome inhibitor.

[0112] In some embodiments, the subjects are those who have relapsed or are refractory to CD38 antibody treatment after remission. An exemplary anti-CD38 antibody is, but is not limited to, daratumumab. In some embodiments, anti-CD38 antibody treatment is monotherapy. In some embodiments, anti-CD38 antibody treatment is part of a combination therapy.

[0113] In some embodiments, subjects have relapsed or are refractory to each of the following: (1) ASCT if eligible to undergo ASCT, (2) at least two consecutive cycles of treatment with an immunomodulator 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, subjects have relapsed or are refractory to each of the following: (1) ASCT if eligible to undergo ASCT, (2) at least two consecutive cycles of treatment with a combination of an immunomodulator and a proteasome inhibitor, and (3) treatment with an anti-CD38 antibody.

[0114] In some embodiments, the subjects are refractory to the last line of previous therapy administered prior to leukocyte apheresis.

[0115] In some embodiments, subjects prior to leukocyte apheresis do not have a history of MM activity infiltrating the central nervous system (CNS). In some embodiments, subjects prior to leukocyte apheresis do not have a history of MM infiltrating the CNS.

[0116] In some embodiments, subjects prior to leukocyte apheresis do not have active plasma cell leukemia, Waldenström macroglobulinemia, polyneuritis, organomegaly, endocrine disorders, monoclonal protein, cutaneous symptom (POEMS) syndrome, any clinically significant amyloidosis, or any combination of the above. In some embodiments, subjects prior to leukocyte apheresis do not have a history of plasma cell leukemia, Waldenström macroglobulinemia, POEMS syndrome, any clinically significant amyloidosis, or any combination of the above. In some embodiments, subjects prior to or before administration of manipulated cells do not have active plasma cell leukemia, Waldenström macroglobulinemia, POEMS syndrome, any clinically significant amyloidosis, or any combination of the above.

[0117] In some embodiments, the subject has not previously received CAR T-cell therapy prior to the administration of BCMA-targeted modified CAR T cells by the method provided. In some embodiments, the subject has not received genetically modified T-cell therapy prior to leukocyte apheresis. In some embodiments, the subject has not received BCMA-targeted therapy prior to leukocyte apheresis. Exemplary BCMA-targeted therapies include, but are not limited to, bispecific T-cell engagement antibodies or molecules, antibody-drug conjugates (BCMA-ADCs), and BCMA-targeted T-cell therapies (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.

[0118] In some aspects, subjects have not received a therapeutic dose of corticosteroids within 14 days prior to leukocyte apheresis. In some aspects, a therapeutic dose of corticosteroids is defined as prednisone or its equivalent greater than 20 mg / day. In some aspects, subjects have not received anti-MM antibodies within 14 days prior to leukocyte apheresis. In some aspects, subjects have not received any other approved systemic anti-MM therapy within 14 days prior to leukocyte apheresis. In some aspects, subjects have not received any experimental therapy within 14 days (biologics) or 5 half-lives (small molecules) prior to leukocyte apheresis. In some aspects, subjects have not undergone autologous stem cell transplantation (SCT) within 6 months prior to leukocyte apheresis. In some aspects, subjects have not undergone allogeneic SCT within 6 months prior to leukocyte apheresis. In some aspects, subjects have not received donor lymphocyte infusion within 6 weeks prior to leukocyte apheresis. In some embodiments, subjects have not received immunosuppressive therapy within four weeks prior to leukocyte apheresis. Examples of immunosuppressive therapies include, but are not limited to, calcineurin inhibitors, methotrexate or other chemotherapeutic agents, mycophenolates, rapamycin, and immunosuppressive antibodies, such as anti-TNF, anti-IL6, or anti-IL6R. In some embodiments, subjects have not received plasmapheresis within 14 days prior to leukocyte apheresis. In some embodiments, subjects have not received radiotherapy targeting a compartment containing a large bone marrow region (e.g., pelvis or sternum) within six weeks prior to leukocyte apheresis. In some embodiments, subjects have not received radiotherapy for a single lesion within 14 days prior to leukocyte apheresis.

[0119] In some embodiments, the eligibility of a subject for a procedure involving the administration of manipulated cells is determined prior to leukocyte apheresis. In some embodiments, the subject prior to leukocyte apheresis has sufficient vascular access for leukocyte apheresis. In some embodiments, the subject prior to leukocyte apheresis 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 to leukocyte apheresis has recovered from any non-hematological toxicity to a grade less than 1 or equal to 1 after the preceding therapy. In some embodiments, the subject prior to leukocyte apheresis has recovered from any non-hematological toxicity to baseline after the preceding therapy.

[0120] In some aspects, subjects prior to leukocyte apheresis have adequate organ function. In some aspects, adequate organ function is, among other factors, 1.0 × 10⁶ without growth factor support within 7 days of eligibility determination. 9 More than cells / L or 1.0 × 10⁻⁶ 9 Absolute neutrophil count (ANC) equal to cells / L; 1.0 × 10⁶ without growth factor support within 14 days of eligibility determination if pegfilgrastim has been previously administered. 9 More than cells / L or 1.0 × 10⁻⁶ 9 ANC equal to cells / L; hemoglobin level higher than or equal to 8 g / dL without red blood cell (RBC) transfusion within 21 days of eligibility determination; 50 × 10 without transfusion within 7 days of eligibility determination. 9Platelet count greater than cells / L; calculated creatinine clearance rate (serum CrCl, Cockcroft-Gault formula) greater than or equal to 60 mL / min without hydration support within 3 days of eligibility determination; aspartate aminotransferase (AST) level less than or equal to 3.0 times the upper limit of normal (ULN); alanine aminotransferase (ALT) level less than or equal to 3.0 times the ULN; total bilirubin level less than 1.5 times the ULN; direct bilirubin level less than 1.5 times the ULN in cases of Gilbert's syndrome; international normalized ratio (INR) less than or equal to 1.5 times the ULN; partial thromboplastin time (PTT) less than or equal to 1.5 times the ULN; e.g., CTCAE grade 1 Adequate pulmonary function is indicated by dyspnea on room air and / or oxygen saturation (SaO2 higher than 92%) or equal to CTCAE grade 1; adequate cardiac function is indicated by left ventricular ejection fraction (LVEF) higher than or equal to 40% as assessed by echocardiography (ECHO) or multi-gate acquisition (MUGA) scan performed within 8 weeks of eligibility determination; or any combination thereof. Adequate organ function may also be indicated by a calculated creatinine clearance rate (CrCl) higher than or equal to 60 mL / min as measured in 24-hour urine collection without hydration support within 3 days of eligibility determination; and / or prothrombin time (PT) lower than or equal to 1.5 times ULN.

[0121] In certain embodiments, prior to the administration of a dose of BCMA-targeted manipulated CAR T cells, the subject is receiving or has received lymphocyte depletion chemotherapy.

[0122] Lymphocyte depletion can improve CAR T cell engraftment and activity through homeostatic cytokines, reduction of CD4+CD25+ regulatory T cells, increase of SDF-1 in the bone marrow microenvironment, and stimulating 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). Additionally, LD chemotherapy may further reduce the tumor burden on the target patient and potentially lower the risk and severity of cytokine release syndrome (CRS).

[0123] Therefore, in some embodiments, the method includes administering a preconditioning agent, such as a lymphocyte depletion agent, or a chemotherapeutic agent, such as cyclophosphamide, fludarabine, or a combination thereof, to a subject prior to administration of the manipulated cells. For example, the subject may be administered the preconditioning agent at least two days before administration of the manipulated cells, for example, at least three, four, five, six, seven, eight, or nine days prior. In some embodiments, the subject is administered the preconditioning agent nine days or less before administration of the manipulated cells, for example, eight, seven, six, five, four, three, or two days or less prior.

[0124] In some embodiments, subjects are preconditioned with cyclophosphamide at a dose of 20 mg to 100 mg or approximately 20 mg to 100 mg per kg of body weight, for example, 40 mg to 80 mg or approximately 40 mg to 80 mg. In some embodiments, subjects are preconditioned with or administered cyclophosphamide at 60 mg / kg or approximately 60 mg / kg. In some embodiments, cyclophosphamide may be administered in a single dose or in multiple doses, for example, given daily, every other day or every three days. In some embodiments, cyclophosphamide is administered once daily over one or two days. In some embodiments, if the lymphocyte depletion agent contains cyclophosphamide, subjects are measured by the body surface area of ​​the subject, including both values, per m². 2 The patient is administered cyclophosphamide in doses of 100mg to 500mg or approximately 100mg to 500mg per unit area, for example, 200mg to 400mg or approximately 200mg to 400mg, or 250mg to 350mg or approximately 250mg to 350mg. In some cases, the patient was given approximately 100mg / m². 2 The patient is administered cyclophosphamide. In some cases, the patient receives approximately 150 mg / m². 2 The patient is administered cyclophosphamide. In some cases, the patient receives approximately 200 mg / m². 2 The patient is administered cyclophosphamide. In some cases, the patient receives approximately 250 mg / m². 2 The patient is administered cyclophosphamide. In some cases, the patient receives approximately 300 mg / m². 2 The subject is administered cyclophosphamide. In some embodiments, cyclophosphamide may be administered in a single dose or in multiple doses, for example, daily, every other day or every three days. In some embodiments, cyclophosphamide is administered daily, for example for 1 to 5 days, or for example for 3 to 5 days. In some cases, the subject has a body surface area of ​​1 m². 2 Approximately 300 mg of cyclophosphamide is administered daily for 3 days prior to the initiation of cell therapy. In some aspects, the subjects receive a total of 300 mg / m².2 , 400 mg / m² 2 500 mg / m² 2 , 600 mg / m² 2 700 mg / m² 2 , 800 mg / m² 2 900 mg / m² 2 , 1000 mg / m² 2 , 1200 mg / m² 2 , 1500 mg / m² 2 , 1800 mg / m² 2 , 2000 mg / m² 2 , 2500 mg / m² 2 , 2700 mg / m² 2 , 3000 mg / m² 2 , 3300 mg / m² 2 , 3600 mg / m² 2 , 4000 mg / m² 2 Alternatively, 5000 mg / m² 2 , or within the range defined by any of the above, or approximately 300 mg / m² 2 , 400 mg / m² 2 500 mg / m² 2 , 600 mg / m² 2 700 mg / m² 2 , 800 mg / m² 2 900 mg / m² 2 , 1000 mg / m² 2 , 1200 mg / m² 2 , 1500 mg / m² 2 , 1800 mg / m² 2 , 2000 mg / m² 2 , 2500 mg / m² 2 , 2700 mg / m² 2 , 3000 mg / m² 2 , 3300 mg / m² 2 , 3600 mg / m² 2 , 4000 mg / m² 2 Alternatively, 5000 mg / m² 2 or cyclophosphamide within the range defined by any of the above is administered before the initiation of cell therapy.

[0125] In some embodiments, if the lymphocyte depletion agent contains fludarabine, the subject is 1 mg / m², including both end values. 2Or approximately 1 mg / m² 2 From 100 mg / m² 2 Or approximately 100 mg / m² 2 For example, 10 mg / m² 2 Or approximately 10 mg / m² 2 From 75 mg / m² 2 Or approximately 75 mg / m² 2 , 15 mg / m² 2 Or approximately 15 mg / m² 2 From 50 mg / m² 2 Or approximately 50 mg / m² 2 , 20 mg / m² 2 Or approximately 20 mg / m² 2 From 40 mg / m² 2 Or approximately 40 mg / m² 2 , 24 mg / m² 2 Or approximately 24 mg / m² 2 From 35 mg / m² 2 Or approximately 35 mg / m² 2 The patient is administered a dose of fludarabine. In some cases, the patient was given 10 mg / m². 2 or 10 mg / m² 2 Or approximately 10 mg / m² 2 Fludarabine is administered. In some cases, the subjects were given 15 mg / m². 2 Or approximately 15 mg / m² 2 Fludarabine is administered. In some cases, the subject was given 20 mg / m². 2 Or approximately 20 mg / m² 2 Fludarabine is administered. In some cases, the subjects were given 25 mg / m². 2 Or approximately 25 mg / m² 2 Fludarabine is administered. In some cases, the subjects were given 30 mg / m². 2 Or approximately 30 mg / m² 2 Fludarabine is administered. In some embodiments, fludarabine may be administered as a single dose or in multiple doses, for example, daily, every other day or every three days. In some embodiments, fludarabine is administered daily, for example for 1 to 5 days, or for example for 3 to 5 days. In some cases, the subject has a body surface area of ​​1 m². 2Fludarabine is administered at a dose of 30 mg or approximately 30 mg per day for 3 days prior to the initiation of cell therapy. In some embodiments, the subject receives a total dose of 10 mg / m². 2 , 20 mg / m² 2 , 25 mg / m² 2 , 30 mg / m² 2 , 40 mg / m² 2 50 mg / m² 2 , 60 mg / m² 2 70 mg / m² 2 , 80 mg / m² 2 90 mg / m² 2 , 100 mg / m² 2 , 120 mg / m² 2 , 150 mg / m² 2 , 180 mg / m² 2 , 200 mg / m² 2 , 250 mg / m² 2 , 270 mg / m² 2 , 300 mg / m² 2 , 330 mg / m² 2 , 360 mg / m² 2 , 400 mg / m² 2 Alternatively, 500 mg / m² 2 , or the range defined by any of the above, or approximately 10 mg / m² 2 , 20 mg / m² 2 , 25 mg / m² 2 , 30 mg / m² 2 , 40 mg / m² 2 50 mg / m² 2 , 60 mg / m² 2 70 mg / m² 2 , 80 mg / m² 2 90 mg / m² 2 , 100 mg / m² 2 , 120 mg / m² 2 , 150 mg / m² 2 , 180 mg / m² 2 , 200 mg / m² 2 , 250 mg / m² 2 , 270 mg / m² 2 , 300 mg / m² 2 , 330 mg / m² 2 , 360 mg / m² 2 , 400 mg / m² 2 Alternatively, 500 mg / m² 2or cyclophosphamide within the range defined by any of the above is administered before the initiation of cell therapy.

[0126] In some embodiments, the lymphocyte depletion agent comprises a single drug, such as cyclophosphamide or fludarabine. In some embodiments, the subject is administered cyclophosphamide alone, without fludarabine or other lymphocyte depletion agents. In some embodiments, prior to administration, the subject receives 200-400 mg or approximately 200-400 mg / m². 2 The target body surface area is arbitrarily 300 or approximately 300 mg / m². 2 The subject is receiving lymphocyte depletion therapy, which includes daily administration of cyclophosphamide for 2 to 4 days. In some embodiments, the subject is administered fludarabine alone, without cyclophosphamide or other lymphocyte depletion agents. In some embodiments, prior to administration, the subject is given 20 to 40 or approximately 20 to 40 mg / m². 2 The target body surface area can be any 30 or approximately 30 mg / m². 2 They are undergoing lymphocyte depletion therapy, which includes daily administration of fludarabine for 2 to 4 days.

[0127] In some embodiments, the lymphocyte depletion agent includes a combination of drugs, for example, a combination of cyclophosphamide and fludarabine. Therefore, the drug combination may include cyclophosphamide in any dose or administration schedule, such as those described above, and fludarabine in any dose or administration schedule, such as those described above. For example, in some aspects, the target is 60 mg / kg or about 60 mg / kg (about 2 g / m²). 2 ) Cyclophosphamide and 3-5 doses of 25 mg / m² before the first or subsequent doses 2 Fludarabine is administered. In some cases, the subjects are given fludarabine (30 mg / m²). 2 ( / day, 3 days) and cyclophosphamide (300 mg / m²) 2(flu / cy) is administered intravenously in parallel for 3 days, prior to cell administration. In some embodiments, subjects are administered one or more doses of lymphocyte depletion agents at reduced, delayed, or eliminated doses.

[0128] In some embodiments, after cells have been collected from a subject and before lymphocyte depletion (LD) chemotherapy is administered, the subject may receive bridging therapy for disease control. Any of the various therapies may be administered as part of bridging therapy to treat a particular disease or condition, based on the judgment of those skilled in the art, including factors based on, for example, the patient's age, the severity or extent of the disease, the possibility of side effects, the timing of administration before LD chemotherapy, preceding therapies, and other factors. In some embodiments, bridging therapy is administered over a period of four weeks or less. Exemplary therapies that may be given as a bridge before 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 approved daratumumab-containing regimens. In some embodiments, bridging therapy is discontinued at least 14 days before LD therapy. In some embodiments, bridging therapy is discontinued 1, 2, 3, 4, 5, 7, 10, 14, 21, 28, 45, or 60 days before lymphocyte depletion. In some embodiments, subjects must recover from bridging therapy-related toxicity to grade 2 or lower before LD chemotherapy.

[0129] In some embodiments, the subject is pre-medicated, for example, to minimize the risk of infusion reaction. In some aspects, the pre-medication includes the step of administering an analgesic and / or antihistamine. In some embodiments, the pre-medication includes the step of administering acetaminophen and / or diphenhydramine, or another H1 antihistamine. In some embodiments, the patient is administered acetaminophen (e.g., 650 mg, orally) and diphenhydramine (e.g., 25-50 mg, IV or orally), or another H1 antihistamine, at the time of treatment in cell therapy or about 30-60 minutes prior.

[0130] In some embodiments, the subjects are at least 18 years of age. In any embodiment of the provided method, the subjects are human subjects.

[0131] A. Medication In some embodiments, the dose of the manipulated cells is administered to the subject according to the method provided and / or together with the provided product or composition. In some embodiments, the size or timing of the dose is determined as a function of the particular disease or condition in the subject. In some cases, the size or timing of the dose for a particular disease, taking into account the provided description, may be determined empirically.

[0132] 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 the extent that it prevents an 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 different but equivalent treatments. For example, in adoptive cell therapy using cells expressing a CAR containing a provided anti-BCMA antibody, the degree of immunogenicity is reduced in some embodiments compared to CARs containing similar, e.g., different antibodies that bind to overlapping epitopes and / or antibodies, e.g., mouse, monkey, rabbit, or humanized antibodies, competing for binding to BCMA.

[0133] In some embodiments, the method comprises adoptive cell therapy, thereby administering genetically engineered cells expressing a provided recombinant receptor containing a BCMA-binding molecule (e.g., a CAR comprising an anti-BCMA antibody or its antigen-binding fragment) to a target. Such administration can promote cell activation (e.g., T cell activation) in a BCMA-targeted manner, and as a result, diseased or impaired cells are targeted for degradation.

[0134] Therefore, the methods and uses provided include methods and uses for adoptive cell therapy. In some embodiments, the methods include the administration of cells or compositions containing cells to a subject, tissue, or cell, e.g., a subject, tissue, or cell that has, is at risk of having, or is suspected of having, a disease, condition, or disorder. In some embodiments, cells, populations, and compositions are administered to a subject having a specific disease or condition to be treated by adoptive cell therapy, e.g., adoptive T-cell therapy. In some embodiments, cells or compositions are administered to a subject, e.g., a subject having, or at risk of having, a disease or condition. In some aspects, the methods thereby treat one or more symptoms of a disease or condition, e.g., remission, such as reducing the tumor burden in BCMA-expressing cancer.

[0135] Methods for administering cells for adoptive cell therapy are publicly known and can be used in connection with the methods and compositions provided. For example, methods for adoptive T cell therapy are described, for example, in U.S. Patent Application Publication No. 2003 / 0170238 by Gruenberg et al., U.S. Patent No. 4,690,915 by Rosenberg, and Rosenberg (2011) Nat Rev Clin Oncol. 8(10):577-85). See, for example, Themeli et al. (2013) Nat Biotechnol. 31(10):928-933; Tsukahara et al. (2013) Biochem Biophys Res Commun 438(1):84-9; and Davila et al. (2013) PLoS ONE 8(4):e61338.

[0136] In some embodiments, cell therapy, such as adoptive cell therapy, such as adoptive T cell therapy, is performed by autologous transplantation, in which case the cells are isolated and / or otherwise prepared from the subject to receive cell therapy or from a sample derived from such a subject. Thus, in some aspects, the cells originate from the subject requiring treatment, such as a patient, and the cells are administered to the same subject after isolation and processing.

[0137] In some embodiments, cell therapy, e.g., adoptive cell therapy, e.g., adoptive T cell therapy, is performed by allogeneic transplantation, in which case the cells are isolated and / or otherwise prepared from a subject other than the subject that will receive or ultimately receive the cell therapy, e.g., a first subject. In such embodiments, the cells are then administered to a different subject of the same species, e.g., a second subject. In some embodiments, the first and second subjects are genetically identical. In some embodiments, the first and second subjects are genetically similar. In some embodiments, the second subject expresses the same HLA class or supertype as the first subject.

[0138] In some embodiments, the subjects to whom the cells, cell populations, or compositions are administered are primates, e.g., humans. In some embodiments, the subjects to whom the cells, cell populations, or compositions are administered are non-human primates. In some embodiments, the non-human primates are monkeys (e.g., cynomolgus macaques) or great apes. The subjects may be male or female and may be of any preferred age, including infants, juveniles, adolescents, adults, and elderly subjects. In some embodiments, the subjects may be non-primate mammals, e.g., rodents (e.g., mice, rats, etc.). In some examples, the patient or subject is a validated animal model for evaluating disease, adoptive cell therapy, and / or toxicity outcomes, e.g., cytokine release syndrome (CRS).

[0139] BCMA-binding molecules such as recombinant receptors (e.g., CARs) and cells expressing them may be administered by any preferred means, for example, by injection, such as intravenous or subcutaneous injection, intraocular injection, periorbital injection, subretinal injection, intravitreous injection, transseptal injection, subscleral injection, intrachoroidal injection, anterior chamber injection, subconjunctival injection, subconjunctival injection, sub-Tenon's capsule injection, retrobulbar injection, peribulbar injection, or delivery near the posterior sclera. In some embodiments, they are administered parenterally, intrapulmonaryly, and intranasally, and intrafocally if desired for local treatment. Parenteral infusions include intramuscular, intravenous, intra-arterial, intraperitoneal, intracranial, intrathoracic, or subcutaneous administration. Dosage and administration may depend in part on whether the administration is short-term or chronic. Various dosing schedules include, but are not limited to, single or multiple doses, bolus administration, and pulse infusion at various points in time.

[0140] For the prevention or treatment of disease, the appropriate dosage of binding molecules, recombinant receptors, or cells may depend on the type of disease being 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, prior therapies, the patient's clinical history and response to the recombinant receptor or cells, and the discretion of the attending physician. The compositions and molecules and cells are preferably administered to the patient in a single dose or over a series of treatments in several embodiments.

[0141] In some embodiments, the method includes the step of administering a dose of engineered cells or a composition containing a dose of engineered cells. In some embodiments, the engineered cells or a composition containing engineered cells may be used in a treatment regimen, which includes the step of administering a dose of engineered cells or a composition containing a dose of engineered cells. In some embodiments, the dose may contain, for example, a specific number or range of recombinant receptor-expressing T cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), for example, any number of such cells described herein. In some embodiments, a composition containing a dose of cells may be administered. In some aspects, the number, amount, or proportion of CAR-expressing (CAR+) cells in a cell population or cell composition may be assessed by detection of surrogate markers, for example by flow cytometry or other means, or by detection of binding of labeled molecules that can specifically bind to binding molecules or receptors provided herein, for example, labeled antigens.

[0142] In any embodiment of the provided, a dose of T cells, e.g., engineered T cells expressing a BCMA-directed CAR, comprises a cell composition or cell population enriched with or containing CD3+ T cells, CD4+ T cells, CD8+ T cells, or CD4+ T cells and CD8+ T cells. In any embodiment of the provided, more than 70%, 75%, 80%, 85%, 90%, 95%, or 98% of the cells in the dose of T cells, or about 70%, 75%, 80%, 85%, 90%, 95%, or 98%, are CD3+ T cells, CD4+ T cells, CD8+ T cells, or CD4+ T cells and CD8+ T cells. In some of any such embodiments, more than 70%, 75%, 80%, 85%, 90%, 95%, or 98% or about 70%, 75%, 80%, 85%, 90%, 95%, or 98% of the cells in the T cell dose are CD3+ T cells. In some of any provided embodiments, the T cell dose includes both CD4+ cells and CD8+ cells. In some of any such embodiments, more than 70%, 75%, 80%, 85%, 90%, 95%, or 98% or about 70%, 75%, 80%, 85%, 90%, 95%, or 98% of the cells in the T cell dose are CD4+ T cells and CD8+ T cells.

[0143] In some embodiments, the cell dose is 0.1 × 10⁶ per kilogram of body weight of the subject. 5 individual or approximately 0.1 × 10 5 From individual cells (cells / kg) to 2 × 10 6 cells / kg or approximately 2 × 10⁻⁶ 6 cells / kg, e.g., 0.1 × 10⁻⁶ 5 cells / kg or approximately 0.1 × 10⁻⁶ 5 Cells / kg to 0.5 × 10 5 cells / kg or approximately 0.5 × 10⁻⁶ 5 cells / kg, 0.5×10 5 cells / kg or approximately 0.5 × 10⁻⁶ 5 From cells / kg to 1 × 10 5 cells / kg or approximately 1 x 10⁻⁶ 5 cells / kg, 1×10 5cells / kg or approximately 1 x 10⁻⁶ 5 From cells / kg: 1.5 × 10 5 Cells / kg or approximately 1.5 × 10⁻⁶ 5 cells / kg, 1.5×10 5 Cells / kg or approximately 1.5 × 10⁻⁶ 5 From cells / kg to 2 × 10 5 cells / kg or approximately 2 x 10⁻⁶ 5 cells / kg, 2×10 5 cells / kg or approximately 2 x 10⁻⁶ 5 2.5 × 10⁻⁶ cells / kg 5 Cells / kg or approximately 2.5 × 10⁻⁶ 5 cells / kg, 2.5×10 5 Cells / kg or approximately 2.5 × 10⁻⁶ 5 3 × 10 cells / kg 5 cells / kg or approximately 3 x 10⁻⁶ 5 cells / kg, 3×10 5 cells / kg or approximately 3 x 10⁻⁶ 5 3.5 × 10⁻⁶ cells / kg 5 Cells / kg or approximately 3.5 × 10⁻⁶ 5 cells / kg, 3.5×10 5 Cells / kg or approximately 3.5 × 10⁻⁶ 5 4 × 10 cells / kg 5 cells / kg or approximately 4 x 10⁻⁶ 5 cells / kg, 4×10 5 cells / kg or approximately 4 x 10⁻⁶ 5 4.5 × 10⁻⁶ cells / kg 5 cells / kg or approximately 4.5 × 10⁻⁶ 5 cells / kg, 4.5×10 5 cells / kg or approximately 4.5 × 10⁻⁶ 5 5 × 10 cells / kg 5 cells / kg or approximately 5 x 10 5 cells / kg, 5×10 5 cells / kg or approximately 5 x 10 5 5.5 × 10⁻⁶ cells / kg 5 Cells / kg or approximately 5.5 x 10⁻⁶ 5 cells / kg, 5.5×10 5 Cells / kg or approximately 5.5 x 10⁻⁶ 5 6 × 10 cells / kg 5cells / kg or approximately 6 x 10⁻⁶ 5 cells / kg, 6×10 5 cells / kg or approximately 6 x 10⁻⁶ 5 6.5 × 10⁻⁶ cells / kg 5 cells / kg or approximately 6.5 × 10⁻⁶ 5 cells / kg, 6.5×10 5 cells / kg or approximately 6.5 × 10⁻⁶ 5 7 × 10 cells / kg 5 cells / kg or approximately 7 x 10⁻⁶ 5 cells / kg, 7×10 5 cells / kg or approximately 7 x 10⁻⁶ 5 7.5 × 10⁻⁶ cells / kg 5 Cells / kg or approximately 7.5 × 10⁻⁶ 5 cells / kg, 7.5×10 5 Cells / kg or approximately 7.5 × 10⁻⁶ 5 8 x 10 cells / kg 5 cells / kg or approximately 8 x 10⁻⁶ 5 cells / kg, or 8 × 10⁶ 5 cells / kg or approximately 8 x 10⁻⁶ 5 From cells / kg to 10x10 5 cells / kg or approximately 10 x 10 5 The material contains BCMA-targeted CAR-operated cells at a concentration of cells / kg. In some embodiments, the cell dose is 2 × 10⁶ cells per kilogram of body weight of the subject. 5 Less than one cell per kg, for example, 3 × 10⁻⁶ 5 Cells / kg or less, or approximately 3 x 10⁻⁶ 5 Cells / kg or less, 4×10 5 Cells / kg or less or approximately 4 x 10 5 Cells / kg or less, 5×10 5 Cells / kg or less or approximately 5 x 10 5 Cells / kg or less, 6×10 5 Cells / kg or less, or approximately 6 x 10⁻⁶ 5 Cells / kg or less, 7×10 5 Cells / kg or less, or approximately 7 x 10 5 Cells / kg or less, 8×10 5 Cells / kg or less, or approximately 8 x 10 5 Cells / kg or less, 9×10 5 Cells / kg or less, or approximately 9 x 105 Cells / kg or less, 1×10 6 Cells / kg or less, or approximately 1 x 10⁻⁶ 6 Cells / kg or less, or 2 × 10⁻⁶ 6 Cells / kg or less, or approximately 2 x 10⁻⁶ 6 The cells include BCMA-targeted CAR-operated cells at a dose of less than 10⁶ cells / kg. In some embodiments, the cell dose is at least 0.1 × 10⁶ cells per kilogram of body weight of the subject. 5 individual or at least about 0.1 × 10 5 individual or 0.1 × 10 5 individual or approximately 0.1 × 10 5 Individuals (cells / kg), for example, at least 0.2 × 10⁻⁶ 5 cells / kg or at least approximately 0.2 × 10⁻⁶ 5 cells / kg or 0.2 × 10⁻⁶ 5 cells / kg or approximately 0.2 × 10⁻⁶ 5 cells / kg, at least 0.3 × 10⁻⁶ 5 cells / kg or at least approximately 0.3 × 10⁻⁶ 5 cells / kg or 0.3 × 10⁻⁶ 5 cells / kg or approximately 0.3 × 10⁻⁶ 5 cells / kg, at least 0.4 × 10⁻⁶ 5 cells / kg or at least approximately 0.4 × 10⁻⁶ 5 cells / kg or 0.4 × 10⁻⁶ 5 cells / kg or approximately 0.4 × 10⁻⁶ 5 cells / kg, at least 0.5 × 10⁻⁶ 5 cells / kg or at least approximately 0.5 × 10⁻⁶ 5 cells / kg or 0.5 × 10⁻⁶ 5 cells / kg or approximately 0.5 × 10⁻⁶ 5 cells / kg, at least 0.6 × 10⁻⁶ 5 cells / kg or at least approximately 0.6 × 10⁻⁶ 5 cells / kg or 0.6 × 10⁻⁶ 5 cells / kg or approximately 0.6 × 10⁻⁶ 5 cells / kg, at least 0.7 × 10⁻⁶ 5 cells / kg or at least approximately 0.7 × 10⁻⁶ 5 cells / kg or 0.7 × 10⁻⁶ 5 cells / kg or approximately 0.7 × 10⁻⁶5 cells / kg, at least 0.8 × 10⁻⁶ 5 cells / kg or at least approximately 0.8 × 10⁻⁶ 5 cells / kg or 0.8 × 10⁻⁶ 5 cells / kg or approximately 0.8 × 10⁻⁶ 5 cells / kg, at least 0.9 × 10⁻⁶ 5 cells / kg or at least approximately 0.9 × 10⁻⁶ 5 cells / kg or 0.9 × 10⁻⁶ 5 cells / kg or approximately 0.9 × 10⁻⁶ 5 cells / kg, at least 0.1 × 10⁻⁶ 6 cells / kg or at least approximately 0.1 × 10⁻⁶ 6 cells / kg or 0.1 × 10⁻⁶ 6 cells / kg or approximately 0.1 × 10⁻⁶ 6 cells / kg, or at least 0.2 × 10⁶ 6 cells / kg or at least approximately 0.2 × 10⁻⁶ 6 cells / kg or 0.2 × 10⁻⁶ 6 cells / kg or approximately 0.2 × 10⁻⁶ 6 The mixture contains BCMA-targeted CAR-manipulated cells in a quantity of cells / kg. In some embodiments, the number of cells is the number of viable cells expressing BCMA-targeted CARs, such as viable T cells, such as viable CD3+ cells.

[0144] In a particular embodiment, an individual population of cells, or a subtype of cells, is defined as 100,000 or approximately 100,000 to 100 billion or approximately 100 billion cells and / or the amount of those cells per kilogram of body weight of the subject, for example, 100,000 or approximately 100,000 to 50 billion or approximately 50 billion cells (for example, 5 million or approximately 5 million cells, 25 million or approximately 25 million cells, 500 million or approximately 500 million cells, 1 billion or approximately 1 billion cells, 5 billion or approximately 5 billion cells, 20 billion or approximately 20 billion cells). , 30 billion or approximately 30 billion cells, 40 billion or approximately 40 billion cells, or any two of the above values) to 1 million or approximately 1 million to 50 billion or approximately 50 billion cells (for example, 5 million or approximately 5 million cells, 25 million or approximately 25 million cells, 500 million or approximately 500 million cells, 1 billion or approximately 1 billion cells, 5 billion or approximately 5 billion cells, 20 billion or approximately 20 billion cells, 30 billion or approximately 30 billion cells, 40 billion or approximately 40 billion cells) Cells (or a range defined by any two of the above values), for example, 10 million or approximately 10 million to 100 billion or approximately 100 billion cells (for example, 20 million or approximately 20 million cells, 30 million or approximately 30 million cells, 40 million or approximately 40 million cells, 60 million or approximately 60 million cells, 70 million or approximately 70 million cells, 80 million or approximately 80 million cells, 90 million or approximately 90 million cells, 10 billion or approximately 10 billion cells, 25 billion or more (The range is defined by any two of the following values: approximately 25 billion cells, 50 billion or approximately 50 billion cells, 75 billion or approximately 75 billion cells, 90 billion or approximately 90 billion cells), and in some cases, from 100 million or approximately 100 million cells to 50 billion or approximately 50 billion cells (for example, 120 million or approximately 120 million cells, 250 million or approximately 250 million cells, 350 million or approximately 350 million cells, 650 million or approximately 650 million cells,The target is administered a number of cells (800 million or approximately 800 million cells, 900 million or approximately 900 million cells, 3 billion or approximately 3 billion cells, 30 billion or approximately 30 billion cells, 45 billion or approximately 45 billion cells) or between these ranges and / or any value per kilogram of the target's body weight. The dosage may vary depending on the disease or disorder and / or attributes specific to the patient and / or other treatment. In some embodiments, such values ​​refer to the number of recombinant receptor-expressing cells, and in other embodiments, they refer to the number of T cells or total cells in the administered composition. In some embodiments, the number of cells is the number of such cells that are viable cells.

[0145] In some embodiments, the cell dose is a uniform or fixed dose of cells, and consequently, the cell dose is not bound by or based on the subject's body surface area or weight.

[0146] In some embodiments, the dose of genetically engineered cells is 1 × 10⁻⁶ 5 individual or approximately 1 x 10 5 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 1.6 × 10 8 individual or approximately 1.6 x 108 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 1.4 × 10 8 individual or approximately 1.4 x 10 8Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 1.2 × 10 8 individual or approximately 1.2 × 10 8Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 7 individual or approximately 1 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 7 individual or approximately 1 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 7 individual or approximately 1 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 7 individual or approximately 1 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 7 individual or approximately 1 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 7 individual or approximately 1 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 7 individual or approximately 1 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 7 individual or approximately 1 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8Total T cells expressing individual BCMA-directed CARs, 1.5×10 7 cells or approximately 1.5×10 7 cells to 2.4×10 8 cells or approximately 2.4×10 8 cells of total T cells expressing individual BCMA-directed CARs, 1.5×10 7 cells or approximately 1.5×10 7 cells to 2.2×10 8 cells or approximately 2.2×10 8 cells of total T cells expressing individual BCMA-directed CARs, 1.5×10 7 cells or approximately 1.5×10 7 cells to 2.0×10 8 cells or approximately 2.0×10 8 cells of total T cells expressing individual BCMA-directed CARs, 1.5×10 7 cells or approximately 1.5×10 7 cells to 1.8×10 8 cells or approximately 1.8×10 8 cells of total T cells expressing individual BCMA-directed CARs, 1.5×10 7 cells or approximately 1.5×10 7 cells to 1.6×10 8 cells or approximately 1.6×10 8 cells of total T cells expressing individual BCMA-directed CARs, 1.5×10 7 cells or approximately 1.5×10 7 cells to 1.4×10 8 cells or approximately 1.4×10 8 cells of total T cells expressing individual BCMA-directed CARs, 1.5×10 7 cells or approximately 1.5×10 7 cells to 1.2×10 8 cells or approximately 1.2×10 8 cells of total T cells expressing individual BCMA-directed CARs, 1.5×10 7 cells or approximately 1.5×10 7 cells to 1.0×10 8 cells or approximately 1.0×10 8 cells of total T cells expressing individual BCMA-directed CARs, 2×10 7 cells or approximately 2×10 7 cells to 2.4×10 8 cells or approximately 2.4×108 Total T cells expressing individual BCMA-directed CARs, 2 × 10 7 cells or about 2 × 10 7 cells to 2.2 × 10 8 cells or about 2.2 × 10 8 Total T cells expressing individual BCMA-directed CARs, 2 × 10 7 cells or about 2 × 10 7 cells to 2.0 × 10 8 cells or about 2.0 × 10 8 Total T cells expressing individual BCMA-directed CARs, 2 × 10 7 cells or about 2 × 10 7 cells to 1.8 × 10 8 cells or about 1.8 × 10 8 Total T cells expressing individual BCMA-directed CARs, 2 × 10 7 cells or about 2 × 10 7 cells to 1.6 × 10 8 cells or about 1.6 × 10 8 Total T cells expressing individual BCMA-directed CARs, 2 × 10 7 cells or about 2 × 10 7 cells to 1.4 × 10 8 cells or about 1.4 × 10 8 Total T cells expressing individual BCMA-directed CARs, 2 × 10 7 cells or about 2 × 10 7 cells to 1.2 × 10 8 cells or about 1.2 × 10 8 Total T cells expressing individual BCMA-directed CARs, 2 × 10 7 cells or about 2 × 10 7 cells to 1.0 × 10 8 cells or about 1.0 × 10 8 Total T cells expressing individual BCMA-directed CARs, 2.5 × 10 7 cells or about 2.5 × 10 7 cells to 2.4 × 10 8 cells or about 2.4 × 10 8 Total T cells expressing individual BCMA-directed CARs, 2.5 × 10 7 cells or about 2.5 × 10 7 cells to 2.2 × 10 8 cells or about 2.2 × 10 8Total T cells expressing BCMA-targeted CARs, 2.5 × 10⁶ 7 1 or approximately 2.5 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 2.5 × 10⁶ 7 1 or approximately 2.5 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 2.5 × 10⁶ 7 1 or approximately 2.5 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 2.5 × 10⁶ 7 1 or approximately 2.5 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 2.5 × 10⁶ 7 1 or approximately 2.5 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 2.5 × 10⁶ 7 1 or approximately 2.5 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 3 × 10⁶ 7 individual or approximately 3 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 3 × 10⁶ 7 individual or approximately 3 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 3 × 10⁶ 7 individual or approximately 3 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 108 Total T cells expressing BCMA-targeted CARs, 3 × 10⁶ 7 individual or approximately 3 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 3 × 10⁶ 7 individual or approximately 3 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 3 × 10⁶ 7 individual or approximately 3 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 3 × 10⁶ 7 individual or approximately 3 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 3 × 10⁶ 7 individual or approximately 3 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 3.5 × 10⁶ 7 1 or approximately 3.5 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 3.5 × 10⁶ 7 1 or approximately 3.5 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 3.5 × 10⁶ 7 1 or approximately 3.5 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 3.5 × 10⁶ 7 1 or approximately 3.5 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 108 Total T cells expressing individual BCMA-targeted CARs, 3.5 × 10 7 1 or approximately 3.5 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 3.5 × 10⁶ 7 1 or approximately 3.5 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 3.5 × 10⁶ 7 1 or approximately 3.5 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 3.5 × 10⁶ 7 1 or approximately 3.5 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 4 × 10 7 individual or approximately 4 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 4 × 10 7 individual or approximately 4 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 4 × 10 7 pieces or approximately 4 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 4 × 10 7 pieces or approximately 4 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 4 × 10 7 individual or approximately 4 x 10 7 From 1.6 × 10 8individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 4 × 10 7 individual or approximately 4 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 4 × 10 7 individual or approximately 4 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 4 × 10 7 individual or approximately 4 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 4.5 × 10⁶ 7 1 or approximately 4.5 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 4.5 × 10⁶ 7 1 or approximately 4.5 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 4.5 × 10⁶ 7 1 or approximately 4.5 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 4.5 × 10⁶ 7 1 or approximately 4.5 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 4.5 × 10⁶ 7 1 or approximately 4.5 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 4.5 × 10⁶ 7 1 or approximately 4.5 x 10 7 From 1.4 × 108 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 4.5 × 10⁶ 7 1 or approximately 4.5 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 4.5 × 10⁶ 7 1 or approximately 4.5 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 7 pieces or approximately 5 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 7 pieces or approximately 5 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 7 pieces or approximately 5 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 7 pieces or approximately 5 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 7 pieces or approximately 5 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 7 pieces or approximately 5 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 7 pieces or approximately 5 x 10 7 From 1.2 × 10 8individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 7 pieces or approximately 5 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 5.5 × 10⁶ 7 1 or approximately 5.5 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 5.5 × 10⁶ 7 1 or approximately 5.5 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 5.5 × 10⁶ 7 1 or approximately 5.5 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 5.5 × 10⁶ 7 1 or approximately 5.5 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 5.5 × 10⁶ 7 1 or approximately 5.5 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 5.5 × 10⁶ 7 1 or approximately 5.5 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 5.5 × 10⁶ 7 1 or approximately 5.5 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 5.5 × 10⁶ 7 1 or approximately 5.5 x 10 7 From 1.0 × 108 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 6 × 10⁶ 7 pieces or approximately 6 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 6 × 10⁶ 7 pieces or approximately 6 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 6 × 10⁶ 7 pieces or approximately 6 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 6 × 10⁶ 7 pieces or approximately 6 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 6 × 10⁶ 7 pieces or approximately 6 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 6 × 10⁶ 7 pieces or approximately 6 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 6 × 10⁶ 7 pieces or approximately 6 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 6 × 10⁶ 7 pieces or approximately 6 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 6.5 × 10⁶ 7 1 or approximately 6.5 x 10 7 From 2.4 × 10 81 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 6.5 × 10⁶ 7 1 or approximately 6.5 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 6.5 × 10⁶ 7 1 or approximately 6.5 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 6.5 × 10⁶ 7 1 or approximately 6.5 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 6.5 × 10⁶ 7 1 or approximately 6.5 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 6.5 × 10⁶ 7 1 or approximately 6.5 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 6.5 × 10⁶ 7 1 or approximately 6.5 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 6.5 × 10⁶ 7 1 or approximately 6.5 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 7 × 10 7 pieces or approximately 7 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 7 × 10 7 pieces or approximately 7 x 10 7 From 2.2 × 108 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 7 × 10 7 pieces or approximately 7 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 7 × 10 7 pieces or approximately 7 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 7 × 10 7 pieces or approximately 7 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 7 × 10 7 pieces or approximately 7 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 7 × 10 7 pieces or approximately 7 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 7 × 10 7 pieces or approximately 7 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 7.5 × 10⁶ 7 1 or approximately 7.5 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 7.5 × 10⁶ 7 1 or approximately 7.5 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 7.5 × 10⁶ 7 1 or approximately 7.5 x 10 7 From 2.0 × 108 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 7.5 × 10⁶ 7 1 or approximately 7.5 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 7.5 × 10⁶ 7 1 or approximately 7.5 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 7.5 × 10⁶ 7 1 or approximately 7.5 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 7.5 × 10⁶ 7 1 or approximately 7.5 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 7.5 × 10⁶ 7 1 or approximately 7.5 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing individual BCMA-targeted CARs, 8×10 7 pieces or approximately 8 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 8 × 10 7 pieces or approximately 8 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 8 × 10 7 pieces or approximately 8 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 8 × 10 7 pieces or approximately 8 x 10 7From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 8 × 10 7 pieces or approximately 8 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 8 × 10 7 pieces or approximately 8 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 8 × 10 7 pieces or approximately 8 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 8 × 10 7 pieces or approximately 8 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 8.5 × 10⁶ 7 1 or approximately 8.5 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 8.5 × 10⁶ 7 1 or approximately 8.5 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 8.5 × 10⁶ 7 1 or approximately 8.5 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 8.5 × 10⁶ 7 1 or approximately 8.5 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 8.5 × 10⁶ 7 1 or approximately 8.5 x 10 7From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 8.5 × 10⁶ 7 1 or approximately 8.5 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 8.5 × 10⁶ 7 1 or approximately 8.5 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 8.5 × 10⁶ 7 1 or approximately 8.5 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 9 × 10 7 pieces or approximately 9 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 9 × 10 7 pieces or approximately 9 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 9 × 10 7 pieces or approximately 9 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 9 × 10 7 pieces or approximately 9 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 9 × 10 7 pieces or approximately 9 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 9 × 10 7 pieces or approximately 9 x 10 7From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 9 × 10 7 pieces or approximately 9 x 10 7 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 9 × 10 7 pieces or approximately 9 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 9.5 × 10⁶ 7 1 or approximately 9.5 x 10 7 From 2.4 × 10 8 1 or approximately 2.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 9.5 × 10⁶ 7 1 or approximately 9.5 x 10 7 From 2.2 × 10 8 1 or approximately 2.2 x 10 8 Total T cells expressing BCMA-targeted CARs, 9.5 × 10⁶ 7 1 or approximately 9.5 x 10 7 From 2.0 × 10 8 1 or approximately 2.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 9.5 × 10⁶ 7 1 or approximately 9.5 x 10 7 From 1.8 × 10 8 individual or approximately 1.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 9.5 × 10⁶ 7 1 or approximately 9.5 x 10 7 From 1.6 × 10 8 individual or approximately 1.6 x 10 8 Total T cells expressing BCMA-targeted CARs, 9.5 × 10⁶ 7 1 or approximately 9.5 x 10 7 From 1.4 × 10 8 individual or approximately 1.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 9.5 × 10⁶ 7 1 or approximately 9.5 x 107 From 1.2 × 10 8 individual or approximately 1.2 × 10 8 Total T cells expressing BCMA-targeted CARs, or 9.5 × 10⁶ 7 1 or approximately 9.5 x 10 7 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 The total number of T cells expressing BCMA-targeted CARs is included. In some embodiments, the number of cells is the number of viable cells, e.g., 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.

[0147] In some embodiments, the dose of genetically engineered cells is 1 × 10⁻⁶ 5 pieces or approximately 1 x 10 5 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 The total T cells express BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 5 × 10⁶ 5 pc or approximately 5 x 10 5 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 The total T cells express BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 1 × 10⁶ 6 pieces or approximately 1 x 10 6 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 The total T cells express BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 5 × 10⁶ 6 pc or approximately 5 x 10 6 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 The total T cells express BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 1 × 10⁶ 7 pieces or approximately 1 x 10 7 From 2.4 × 108 pc or approximately 2.4 × 10 8 The total T cells express BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 1.5 × 10⁶ 7 pc or approximately 1.5 × 10 7 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 The total T cells express BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 2 × 10⁶ 7 pc or approximately 2 × 10 7 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 It contains total T cells expressing BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 2.5 × 10⁶ 7 pc or approximately 2.5 × 10 7 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 The total T cells express BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 3 × 10⁶ 7 pieces or approximately 3 x 10 7 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 It contains total T cells expressing BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 3.5 × 10⁶ 7 pc or approximately 3.5 × 10 7 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 It contains total T cells expressing BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 4.5 × 10⁶ 7 pc or approximately 4.5 × 10 7 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 It contains total T cells expressing BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 5.5 × 10⁶ 7 pc or approximately 5.5 × 10 7 From 2.4 × 10 8 pc or approximately 2.4 × 108 The total T cells express BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 6 × 10⁶ 7 pc or approximately 6 x 10 7 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 It contains total T cells expressing BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 6.5 × 10⁶ 7 pc or approximately 6.5 × 10 7 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 The total T cells express BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 7 × 10⁶ 7 pcs or approximately 7 x 10 7 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 The total T cells express BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 7.5 × 10⁶ 7 pc or approximately 7.5 × 10 7 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 The total T cells express BCMA-targeted CARs. In some embodiments, the dose of genetically engineered cells is 8 × 10⁶ 7 pc or approximately 8 x 10 7 From 2.4 × 10 8 pc or approximately 2.4 × 10 8 The total number of T cells expressing BCMA-targeted CARs is included. In some embodiments, the number of cells is the number of viable cells, e.g., 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.

[0148] In some embodiments, the dose of genetically engineered cells is 1 × 10⁻⁶ 5 individual or approximately 1 x 10 5 From 1 × 10 8 individual or approximately 1 x 108 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 0.8 × 10 8 individual or approximately 0.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 0.6 × 10 8 individual or approximately 0.6 × 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 0.4 × 10 8 individual or approximately 0.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 0.2 × 10 8 individual or approximately 0.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 1.0 × 10 7 individual or approximately 1.0 x 10 7 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 0.8 × 10 7 individual or approximately 0.8 x 10 7 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 0.6 × 10 7 individual or approximately 0.6 × 10 7 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 0.4 × 10 7 individual or approximately 0.4 x 10 7 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 0.2 × 10 7 individual or approximately 0.2 × 10 7Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 5 individual or approximately 1 x 10 5 From 1.0 × 10 6 individual or approximately 1.0 x 10 6 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 0.8 × 10 8 individual or approximately 0.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 0.6 × 10 8 individual or approximately 0.6 × 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 0.4 × 10 8 individual or approximately 0.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 0.2 × 10 8 individual or approximately 0.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 1.0 × 10 7 individual or approximately 1.0 x 10 7 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 0.8 × 10 7 individual or approximately 0.8 x 10 7 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 0.6 × 10 7 individual or approximately 0.6 × 10 7Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 0.4 × 10 7 individual or approximately 0.4 x 10 7 Total T cells expressing BCMA-targeted CARs, 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 0.2 × 10 7 individual or approximately 0.2 × 10 7 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 0.8 × 10 8 individual or approximately 0.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 0.6 × 10 8 individual or approximately 0.6 × 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 0.4 × 10 8 individual or approximately 0.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 0.2 × 10 8 individual or approximately 0.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 1.0 × 10 7 individual or approximately 1.0 x 10 7 Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 0.8 × 10 7 individual or approximately 0.8 x 10 7Total T cells expressing BCMA-targeted CARs, 5 × 10 6 pieces or approximately 5 x 10 6 From 0.6 × 10 7 individual or approximately 0.6 × 10 7 Total T cells expressing BCMA-targeted CARs, 10 × 10 6 individual or approximately 10 x 10 6 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 10 × 10 6 individual or approximately 10 x 10 6 From 0.9 × 10 8 individual or approximately 0.9 × 10 8 Total T cells expressing BCMA-targeted CARs, 10 × 10 6 individual or approximately 10 x 10 6 From 0.8 × 10 8 individual or approximately 0.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 10 × 10 6 individual or approximately 10 x 10 6 From 0.7 × 10 8 individual or approximately 0.7 × 10 8 Total T cells expressing BCMA-targeted CARs, 10 × 10 6 individual or approximately 10 x 10 6 From 0.6 × 10 8 individual or approximately 0.6 × 10 8 Total T cells expressing BCMA-targeted CARs, 10 × 10 6 individual or approximately 10 x 10 6 From 0.5 × 10 8 individual or approximately 0.5 x 10 8 Total T cells expressing BCMA-targeted CARs, 10 × 10 6 individual or approximately 10 x 10 6 From 0.4 × 10 8 individual or approximately 0.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 10 × 10 6 individual or approximately 10 x 10 6 From 0.3 × 10 8 individual or approximately 0.3 × 10 8Total T cells expressing BCMA-targeted CARs, 10 × 10 6 individual or approximately 10 x 10 6 From 0.2 × 10 8 individual or approximately 0.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 10 × 10 6 individual or approximately 10 x 10 6 From 15 x 10 6 individual or approximately 15 x 10 6 Total T cells expressing BCMA-targeted CARs, 15 × 10⁶ 6 individual or approximately 15 x 10 6 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 15 × 10⁶ 6 individual or approximately 15 x 10 6 From 0.9 × 10 8 individual or approximately 0.9 × 10 8 Total T cells expressing BCMA-targeted CARs, 15 × 10⁶ 6 individual or approximately 15 x 10 6 From 0.8 × 10 8 individual or approximately 0.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 15 × 10⁶ 6 individual or approximately 15 x 10 6 From 0.7 × 10 8 individual or approximately 0.7 × 10 8 Total T cells expressing BCMA-targeted CARs, 15 × 10⁶ 6 individual or approximately 15 x 10 6 From 0.6 × 10 8 individual or approximately 0.6 × 10 8 Total T cells expressing BCMA-targeted CARs, 15 × 10⁶ 6 individual or approximately 15 x 10 6 From 0.5 × 10 8 individual or approximately 0.5 x 10 8 Total T cells expressing BCMA-targeted CARs, 15 × 10⁶ 6 individual or approximately 15 x 10 6 From 0.4 × 10 8 individual or approximately 0.4 x 10 8Total T cells expressing BCMA-targeted CARs, 15 × 10⁶ 6 individual or approximately 15 x 10 6 From 0.3 × 10 8 individual or approximately 0.3 × 10 8 Total T cells expressing BCMA-targeted CARs, 15 × 10⁶ 6 individual or approximately 15 x 10 6 From 0.2 × 10 8 individual or approximately 0.2 × 10 8 Total T cells expressing BCMA-targeted CARs, 20 × 10 6 1 or approximately 20 x 10 6 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 20 × 10 6 1 or approximately 20 x 10 6 From 0.9 × 10 8 individual or approximately 0.9 × 10 8 Total T cells expressing BCMA-targeted CARs, 20 × 10 6 1 or approximately 20 x 10 6 From 0.8 × 10 8 individual or approximately 0.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 20 × 10 6 1 or approximately 20 x 10 6 From 0.7 × 10 8 individual or approximately 0.7 × 10 8 Total T cells expressing BCMA-targeted CARs, 20 × 10 6 1 or approximately 20 x 10 6 From 0.6 × 10 8 individual or approximately 0.6 × 10 8 Total T cells expressing BCMA-targeted CARs, 20 × 10 6 1 or approximately 20 x 10 6 From 0.5 × 10 8 individual or approximately 0.5 x 10 8 Total T cells expressing BCMA-targeted CARs, 20 × 10 6 1 or approximately 20 x 10 6 From 0.4 × 10 8 individual or approximately 0.4 x 10 8Total T cells expressing BCMA-targeted CARs, 20 × 10 6 1 or approximately 20 x 10 6 From 0.3 × 10 8 individual or approximately 0.3 × 10 8 Total T cells expressing BCMA-targeted CARs, 20 × 10 6 1 or approximately 20 x 10 6 From 25 x 10 6 1 or approximately 25 x 10 6 Total T cells expressing BCMA-targeted CARs, 25 × 10⁶ 6 1 or approximately 25 x 10 6 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 25 × 10⁶ 6 1 or approximately 25 x 10 6 From 0.9 × 10 8 individual or approximately 0.9 × 10 8 Total T cells expressing BCMA-targeted CARs, 25 × 10⁶ 6 1 or approximately 25 x 10 6 From 0.8 × 10 8 individual or approximately 0.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 25 × 10⁶ 6 1 or approximately 25 x 10 6 From 0.7 × 10 8 individual or approximately 0.7 × 10 8 Total T cells expressing BCMA-targeted CARs, 25 × 10⁶ 6 1 or approximately 25 x 10 6 From 0.6 × 10 8 individual or approximately 0.6 × 10 8 Total T cells expressing BCMA-targeted CARs, 25 × 10⁶ 6 1 or approximately 25 x 10 6 From 0.5 × 10 8 individual or approximately 0.5 x 10 8 Total T cells expressing BCMA-targeted CARs, 25 × 10⁶ 6 1 or approximately 25 x 10 6 From 0.4 × 10 8 individual or approximately 0.4 x 10 8Total T cells expressing BCMA-targeted CARs, 25 × 10⁶ 6 1 or approximately 25 x 10 6 From 0.3 × 10 8 individual or approximately 0.3 × 10 8 Total T cells expressing BCMA-targeted CARs, 30 × 10 6 individual or approximately 30 x 10 6 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 30 × 10 6 individual or approximately 30 x 10 6 From 0.9 × 10 8 individual or approximately 0.9 × 10 8 Total T cells expressing BCMA-targeted CARs, 30 × 10 6 individual or approximately 30 x 10 6 From 0.8 × 10 8 individual or approximately 0.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 30 × 10 6 individual or approximately 30 x 10 6 From 0.7 × 10 8 individual or approximately 0.7 × 10 8 Total T cells expressing BCMA-targeted CARs, 30 × 10 6 individual or approximately 30 x 10 6 From 0.6 × 10 8 individual or approximately 0.6 × 10 8 Total T cells expressing BCMA-targeted CARs, 30 × 10 6 individual or approximately 30 x 10 6 From 0.5 × 10 8 individual or approximately 0.5 x 10 8 Total T cells expressing BCMA-targeted CARs, 30 × 10 6 individual or approximately 30 x 10 6 From 0.4 × 10 8 individual or approximately 0.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 30 × 10 6 individual or approximately 30 x 10 6 From 35 x 10 6 Individual or approximately 35 x 10 6Total T cells expressing individual BCMA-targeted CARs, 35×10 6 Individual or approximately 35 x 10 6 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 35 × 10⁶ 6 Individual or approximately 35 x 10 6 From 0.9 × 10 8 individual or approximately 0.9 × 10 8 Total T cells expressing BCMA-targeted CARs, 35 × 10⁶ 6 Individual or approximately 35 x 10 6 From 0.8 × 10 8 individual or approximately 0.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 35 × 10⁶ 6 Individual or approximately 35 x 10 6 From 0.7 × 10 8 individual or approximately 0.7 × 10 8 Total T cells expressing BCMA-targeted CARs, 35 × 10⁶ 6 Individual or approximately 35 x 10 6 From 0.6 × 10 8 individual or approximately 0.6 × 10 8 Total T cells expressing BCMA-targeted CARs, 35 × 10⁶ 6 Individual or approximately 35 x 10 6 From 0.5 × 10 8 individual or approximately 0.5 x 10 8 Total T cells expressing BCMA-targeted CARs, 35 × 10⁶ 6 Individual or approximately 35 x 10 6 From 0.4 × 10 8 individual or approximately 0.4 x 10 8 Total T cells expressing BCMA-targeted CARs, 40 × 10 6 1 or approximately 40 x 10 6 From 1.0 × 10 8 individual or approximately 1.0 x 10 8 Total T cells expressing BCMA-targeted CARs, 40 × 10 6 1 or approximately 40 x 10 6 From 0.9 × 10 8 individual or approximately 0.9 × 108 Total T cells expressing BCMA-targeted CARs, 40 × 10 6 1 or approximately 40 x 10 6 From 0.8 × 10 8 individual or approximately 0.8 x 10 8 Total T cells expressing BCMA-targeted CARs, 40 × 10 6 1 or approximately 40 x 10 6 From 0.7 × 10 8 individual or approximately 0.7 × 10 8 Total T cells expressing BCMA-targeted CARs, 40 × 10 6 1 or approximately 40 x 10 6 From 0.6 × 10 8 individual or approximately 0.6 × 10 8 Total T cells expressing BCMA-targeted CARs, 40 × 10 6 1 or approximately 40 x 10 6 From 0.5 × 10 8 individual or approximately 0.5 x 10 8 Total T cells expressing BCMA-targeted CARs, or 40 × 10⁶ 6 1 or approximately 40 x 10 6 From 45 x 10 6 individual or approximately 45 x 10 6 The total number of T cells expressing BCMA-targeted CARs is included. In some embodiments, the number of cells is the number of such cells, which are viable cells, e.g., viable T cells.

[0149] In some embodiments, the dose of genetically engineered cells is at least 1 × 10⁶ 5 individual or at least approximately 1 × 10 5 T cells expressing BCMA-targeted CARs, at least 2.5 × 10⁶ 5 individual or at least approximately 2.5 × 10 5 T cells expressing BCMA-targeted CARs, at least 5 × 10⁶ 5 individual or at least approximately 5 x 10 5 T cells expressing BCMA-targeted CARs, at least 1 × 10⁶ 6 individual or at least approximately 1 × 10 6T cells expressing BCMA-targeted CARs, at least 2.5 × 10⁶ 6 individual or at least approximately 2.5 × 10 6 T cells expressing BCMA-targeted CARs, at least 5 × 10⁶ 6 individual or at least approximately 5 x 10 6 T cells expressing BCMA-targeted CARs, at least 1 × 10⁶ 7 individual or at least approximately 1 × 10 7 T cells expressing BCMA-targeted CARs, at least 2.5 × 10⁶ 7 individual or at least approximately 2.5 × 10 7 T cells expressing BCMA-targeted CARs, or at least 5 × 10⁶ 7 individual or at least approximately 5 x 10 7 The present invention includes T cells expressing BCMA-targeted CARs. In some embodiments, the number of cells is the number of such cells, which are viable cells, for example, viable T cells.

[0150] In some embodiments, the dose of genetically engineered cells is 1 × 10⁻⁶ 5 Less than one or approximately 1 x 10 5 T cells expressing fewer than BCMA-targeted CARs, 2.5 × 10⁶ 5 Less than one or approximately 2.5 × 10 5 T cells expressing fewer than BCMA-targeted CARs, 5 × 10 5 Less than one or approximately 5 x 10 5 T cells expressing fewer than BCMA-targeted CARs, 1 × 10 6 Less than one or approximately 1 x 10 6 T cells expressing fewer than BCMA-targeted CARs, 2.5 × 10⁶ 6 Less than one or approximately 2.5 × 10 6 T cells expressing fewer than BCMA-targeted CARs, 5 × 10 6 Less than one or approximately 5 x 10 6 T cells expressing fewer than BCMA-targeted CARs, 1 × 10 7 Less than one or approximately 1 x 10 7 T cells expressing fewer than BCMA-targeted CARs, 1.5 × 10⁶ 7 Less than one or approximately 1.5 × 107 T cells expressing fewer than BCMA-targeted CARs, 2 × 10 7 Less than 10 or approximately 2 x 10 7 T cells expressing fewer than BCMA-targeted CARs, 2.5 × 10⁶ 7 Less than one or approximately 2.5 × 10 7 T cells expressing fewer than BCMA-targeted CARs, 3 × 10 7 Less than one or approximately 3 x 10 7 T cells expressing fewer than BCMA-targeted CARs, 3.5 × 10⁶ 7 Less than one or approximately 3.5 × 10 7 T cells expressing fewer than BCMA-targeted CARs, 4 × 10 7 Less than one or approximately 4 x 10 7 T cells expressing fewer than BCMA-targeted CARs, 4.5 × 10⁶ 7 Less than one or approximately 4.5 × 10 7 T cells expressing fewer than BCMA-targeted CARs, or 5 × 10⁶ 7 Less than one or approximately 5 x 10 7 The number of cells includes fewer than 1 T cells expressing BCMA-targeted CARs. In some embodiments, the number of cells is the number of such cells, which are viable cells, e.g., viable T cells.

[0151] In some aspects, cell therapy involves 1 × 10⁶ values, each including the values ​​at both ends. 5 individual or approximately 1 x 10 5 From 1 × 10 8 individual or approximately 1 x 10 8 5 × 10¹ total recombinant receptor-expressing cells or total T cells 5 pieces or approximately 5 x 10 5 From 5 x 10 7 pieces or approximately 5 x 10 7 10⁴ total recombinant receptor-expressing cells or total T cells, or 1 × 10⁶ 6 individual or approximately 1 x 10 6 From 1 × 10 7 individual or approximately 1 x 10 7 The administration of a dose containing a large number of cells, including a total recombinant receptor-expressing cell or total T cells. In some embodiments, the cell therapy includes 1 × 10⁶ values, each including the values ​​at both ends.5 individual or approximately 1 x 10 5 From 1 × 10 8 individual or approximately 1 x 10 8 5 × 10¹ total recombinant receptor-expressing cells, or total T cells 5 pieces or approximately 5 x 10 5 From 1 × 10 8 individual or approximately 1 x 10 8 1 x 10⁶ total recombinant receptor-expressing cells, or total T cells 6 individual or approximately 1 x 10 6 From 50 x 10 6 1 or approximately 50 x 10 6 5 × 10¹ total recombinant receptor-expressing cells, or total T cells 6 pieces or approximately 5 x 10 6 From 45 x 10 6 individual or approximately 45 x 10 6 10 x 10 total recombinant receptor-expressing cells or total T cells, or 10 x 10 6 individual or approximately 10 x 10 6 From 25 x 10 6 1 or approximately 25 x 10 6 The administration of a dose containing a large number of total recombinant receptor-expressing cells or total T cells comprises at least 1 × 10⁶ cells. In some embodiments, the cell therapy comprises at least 1 × 10⁶ cells. 5 individual or at least about 1 × 10 5 A total number of recombinant receptor-expressing cells or total T cells, e.g., at least 1 × 10⁶ 6 individual or at least 1 × 10 6 each, at least 1 × 10 7 individual or at least about 1 × 10 7 each, at least 1 × 10 8 individual or at least about 1 × 10 8 The method involves administering a dose of cells containing a large number of such cells. In some embodiments, the number of cells is the number of such cells that are viable cells, for example, viable T cells.

[0152] In some embodiments, for example, if the subject is human, the dose is 5 × 10 6 pc or approximately 5 x 10 6More than 100 total CAR-expressing (CAR+) cells, T cells, or peripheral blood mononuclear cells (PBMCs) and 100 × 10⁶ 6 1 or approximately 100 x 10 6 The total number of CAR-expressing cells, T cells, or PBMCs is less than 10. In some embodiments, the dose of genetically engineered cells is 5 × 10, including the values ​​at both ends. 6 pieces or approximately 5 x 10 6 From 10x10 6 individual or approximately 10 x 10 6 Total CAR-expressing (CAR+) T cells, 10 × 10 6 individual or approximately 10 x 10 6 From 15 x 10 6 individual or approximately 15 x 10 6 15 × 10¹ CAR+ T cells 6 individual or approximately 15 x 10 6 From 20 x 10 6 1 or approximately 20 x 10 6 10 CAR+ T cells, 20 × 10 6 1 or approximately 20 x 10 6 From 25 x 10 6 1 or approximately 25 x 10 6 10 CAR+ T cells, 25 × 10 6 1 or approximately 25 x 10 6 From 30 x 10 6 individual or approximately 30 x 10 6 CAR+ T cells, 30 × 10 6 individual or approximately 30 x 10 6 From 35 x 10 6 Individual or approximately 35 x 10 6 CAR+ T cells, 35 × 10 6 Individual or approximately 35 x 10 6 From 40 x 10 6 1 or approximately 40 x 10 6 CAR+ T cells, 40 × 10 6 1 or approximately 40 x 10 6 From 45 x 10 6 individual or approximately 45 x 10 6 CAR+ T cells, 45 × 10 6 individual or approximately 45 x 10 6 From 50 x 10 61 or approximately 50 x 10 6 50 x 10¹ CAR+ T cells 6 1 or approximately 50 x 10 6 From 55 x 10 6 individual or approximately 55 x 10 6 CAR+ T cells, 55 × 10 6 individual or approximately 55 x 10 6 From 60 x 10 6 individual or approximately 60 x 10 6 CAR+ T cells, 60 × 10 6 individual or approximately 60 x 10 6 From 65 x 10 6 Individual or approximately 65 x 10 6 CAR+ T cells, 65 × 10 6 Individual or approximately 65 x 10 6 From 70 x 10 6 Individual or approximately 70 x 10 6 CAR+ T cells, 70 × 10 6 Individual or approximately 70 x 10 6 From 75 x 10 6 Individual or approximately 75 x 10 6 CAR+ T cells, 75 × 10 6 Individual or approximately 75 x 10 6 From 80 x 10 6 1 or approximately 80 x 10 6 CAR+ T cells, 80 × 10 6 1 or approximately 80 x 10 6 From 85 x 10 6 individual or approximately 85 x 10 6 CAR+ T cells, 85 × 10 6 individual or approximately 85 x 10 6 From 90 x 10 6 1 or approximately 90 x 10 6 CAR+ T cells, 90 × 10 6 1 or approximately 90 x 10 6 From 95 x 10 6 Individual or approximately 95 x 10 6 10 CAR+ T cells, or 95 × 10⁶ 6 Individual or approximately 95 x 10 6 From 100 x 10 6 1 or approximately 100 x 10 6It contains CAR+ T cells. In any of the preceding embodiments, the CAR+ T cells express BCMA-targeted CARs, such as those derived from BCMA-55.

[0153] In some embodiments, for example, if the subject is human, the dose is 5 × 10 6 pc or approximately 5 x 10 6 It comprises a total recombinant receptor (e.g., CAR) expressing cell, T cell, or peripheral blood mononuclear cell (PBMC). In some embodiments, for example, when the subject is human, the dose is 10 × 10 6 1 or approximately 10 x 10 6 It comprises a total recombinant receptor (e.g., CAR) expressing cell, T cell, or peripheral blood mononuclear cell (PBMC). In some embodiments, for example, when the subject is human, the dose is 20 × 10⁻¹⁶ 6 1 or approximately 20 x 10 6 It comprises a total recombinant receptor (e.g., CAR) expressing cell, T cell, or peripheral blood mononuclear cell (PBMC). In some embodiments, for example, when the subject is human, the dose is 30 × 10 6 1 or approximately 30 x 10 6 It comprises a total recombinant receptor (e.g., CAR) expressing cell, T cell, or peripheral blood mononuclear cell (PBMC). In some embodiments, for example, when the subject is human, the dose is 40 × 10⁻¹⁶ 6 1 or approximately 40 x 10 6 It comprises a total recombinant receptor (e.g., CAR) expressing cell, T cell, or peripheral blood mononuclear cell (PBMC). In some embodiments, for example, when the subject is human, the dose is 60 × 10⁻¹⁶ 6 1 or approximately 60 x 10 6 It comprises a total recombinant receptor (e.g., CAR) expressing cell, T cell, or peripheral blood mononuclear cell (PBMC). In some embodiments, for example, when the subject is human, the dose is 80 × 10⁻¹⁶ 6 1 or approximately 80 x 10 6 This includes a total recombinant receptor (e.g., CAR) expressing cell, T cell, or peripheral blood mononuclear cell (PBMC).

[0154] In some embodiments, this number relates to the total number of CD3+ or CD8+, and in some cases further to the total number of CAR-expressing (e.g., CAR+) cells. In some embodiments, the dose of genetically engineered cells is 1 × 10⁻¹⁶, including the values ​​at both ends. 7 individual or approximately 1 x 10 7 From 1.5 × 10 7 1 or approximately 1.5 x 10 7 Individual CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, 1.5 × 10⁶ 7 1 or approximately 1.5 x 10 7 From 2 × 10 7 individual or approximately 2 x 10 7 2 × 10¹ CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells 7 individual or approximately 2 x 10 7 From 2.5 × 10 7 1 or approximately 2.5 x 10 7 Individual CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, 2.5 × 10⁶ 7 1 or approximately 2.5 x 10 7 From 3 × 10 7 individual or approximately 3 x 10 7 3 × 10¹ CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells 7 individual or approximately 3 x 10 7 From 3.5 × 10 7 1 or approximately 3.5 x 10 7 Individual CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, 3.5 × 10⁶ 7 1 or approximately 3.5 x 10 7 From 4 x 10 7 individual or approximately 4 x 10 7 4 × 10¹ CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells 7 individual or approximately 4 x 10 7 From 4.5 × 10 7 1 or approximately 4.5 x 10 7Individual CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, 4.5 × 10⁶ 7 1 or approximately 4.5 x 10 7 From 5 x 10 7 pieces or approximately 5 x 10 7 5 × 10¹ CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells 7 pieces or approximately 5 x 10 7 From 5.5 × 10 7 1 or approximately 5.5 x 10 7 Individual CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, 5.5 × 10⁶ 7 1 or approximately 5.5 x 10 7 From 6 × 10 7 pieces or approximately 6 x 10 7 6 × 10¹ CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells 7 pieces or approximately 6 x 10 7 From 6.5 × 10 7 1 or approximately 6.5 x 10 7 Individual CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, 6.5 × 10⁶ 7 1 or approximately 6.5 x 10 7 From 7.5 × 10 7 1 or approximately 7.5 x 10 7 10 CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells, or 7.5 × 10 7 1 or approximately 7.5 x 10 7 From 8 x 10 7 pieces or approximately 8 x 10 7 Includes individual CD3+ or CD8+ total T cells or CD3+ or CD8+ CAR-expressing cells.

[0155] In some embodiments, the dose of genetically modified cells is related to the total number of CD3+CAR-expressing (CAR+) or CD4+ / CD8+CAR-expressing (CAR+) cells. In some embodiments, the dose of genetically modified cells is 1 × 10⁻¹⁶, including the values ​​at both ends. 7individual or approximately 1 x 10 7 From 1.5 × 10 7 1 or approximately 1.5 x 10 7 Individual CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, 1.5 × 10⁶ 7 1 or approximately 1.5 x 10 7 From 2 × 10 7 individual or approximately 2 x 10 7 2 × 10¹ CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells 7 individual or approximately 2 x 10 7 From 2.5 × 10 7 1 or approximately 2.5 x 10 7 2.5 × 10¹ CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells 7 1 or approximately 2.5 x 10 7 From 3 × 10 7 individual or approximately 3 x 10 7 3 × 10¹ CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells 7 individual or approximately 3 x 10 7 From 3.5 × 10 7 1 or approximately 3.5 x 10 7 Individual CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, 3.5 × 10⁶ 7 1 or approximately 3.5 x 10 7 From 4 x 10 7 individual or approximately 4 x 10 7 4 × 10¹ CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells 7 individual or approximately 4 x 10 7 From 4.5 × 10 7 1 or approximately 4.5 x 10 7 Individual CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, 4.5 × 10⁶ 7 1 or approximately 4.5 x 10 7From 5 x 10 7 pieces or approximately 5 x 10 7 5 × 10¹ CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells 7 pieces or approximately 5 x 10 7 From 5.5 × 10 7 1 or approximately 5.5 x 10 7 5.5 × 10¹ CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells 7 1 or approximately 5.5 x 10 7 From 6 × 10 7 pieces or approximately 6 x 10 7 6 × 10¹ CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells 7 pieces or approximately 6 x 10 7 From 6.5 × 10 7 1 or approximately 6.5 x 10 7 6.5 × 10¹ CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells 7 1 or approximately 6.5 x 10 7 From 7.5 × 10 7 1 or approximately 7.5 x 10 7 10 CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells, or 7.5 × 10⁶ 7 1 or approximately 7.5 x 10 7 From 8 x 10 7 pieces or approximately 8 x 10 7 Includes individual CD3+ or CD4+ / CD8+ total T cells or CD3+ or CD4+ / CD8+ CAR-expressing cells.

[0156] In some embodiments, the dose is 1.0 × 10 7 , 2.0×10 7 , 3.0×10 7 , 4.0×10 7 , 6.0×10 7 , or 8.0 x 10 7 individual or approximately 1.0 × 10 7, 2.0×10 7 , 3.0×10 7 , 4.0×10 7 , 6.0×10 7 , or 8.0 x 10 7 It contains individual CD3+ or CD4+ / CD8+ total T cells or CD3+CAR-expressing or CD4+ / CD8+CAR-expressing cells. In some embodiments, the dose is 1.0 × 10⁻¹⁶ 7 , 2.0×10 7 , 3.0×10 7 , 4.0×10 7 , 6.0×10 7 , or 8.0 x 10 7 individual or approximately 1.0 × 10 7 , 2.0×10 7 , 3.0×10 7 , 4.0×10 7 , 6.0×10 7 , or 8.0 x 10 7 Contains 10 CD3+CAR-expressing cells. In some embodiments, the dose is 1.0 × 10⁶ 7 , 2.0×10 7 , 3.0×10 7 , 4.0×10 7 , 6.0×10 7 , or 8.0 x 10 7 individual or approximately 1.0 × 10 7 , 2.0×10 7 , 3.0×10 7 , 4.0×10 7 , 6.0×10 7 , or 8.0 x 10 7 Contains individual CD4+ / CD8+CAR-expressing cells.

[0157] In some embodiments, the dose is 0.5 × 10 7 pc or approximately 0.5 × 10 7 From individual CD3+CAR-expressing cells, 1.5 × 10⁶ 7 pc or approximately 1.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 1.5 × 10⁶ 7 pc or approximately 1.5 × 10 7 From individual CD3+CAR-expressing cells, 2.5 × 10⁶ 7pc or approximately 2.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 2.5 × 10⁶ 7 pc or approximately 2.5 × 10 7 From individual CD3+CAR-expressing cells, 3.5 × 10⁶ 7 pc or approximately 3.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 3.5 × 10⁶ 7 pc or approximately 3.5 × 10 7 From individual CD3+CAR-expressing cells, 4.5 × 10⁶ 7 pc or approximately 4.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 4.5 × 10⁶ 7 pc or approximately 4.5 × 10 7 From individual CD3+CAR-expressing cells, 5.5 × 10⁶ 7 pc or approximately 5.5 × 10 7 These are CD3+CAR-expressing cells. In some embodiments, the dose is 5.5 × 10⁶ 7 pc or approximately 5.5 × 10 7 From individual CD3+CAR-expressing cells, 6.5 × 10⁶ 7 pc or approximately 6.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 6.5 × 10⁶ 7 pc or approximately 6.5 × 10 7 From individual CD3+CAR-expressing cells, 7.5 × 10⁶ 7 pc or approximately 7.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 7.5 × 10⁶ 7 pc or approximately 7.5 × 10 7 From individual CD3+CAR-expressing cells, 8.5 × 10⁶ 7 pc or approximately 8.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 8.5 × 10⁶ 7 pc or approximately 8.5 × 10 7 From individual CD3+CAR-expressing cells, 9.5 × 10⁶ 7 pc or approximately 9.5 × 10 7 These are CD3+CAR-expressing cells. In some embodiments, the dose is 9.5 × 10⁶7 pc or approximately 9.5 × 10 7 From individual CD3+CAR-expressing cells, 10.5 × 10⁶ 7 1 or approximately 10.5 x 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 10.5 × 10⁶ 7 1 or approximately 10.5 x 10 7 From individual CD3+CAR-expressing cells, 11.5 × 10⁶ 7 pc or approximately 11.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 11.5 × 10⁶ 7 pc or approximately 11.5 × 10 7 From individual CD3+CAR-expressing cells, 12.5 × 10⁶ 7 Individual or approximately 12.5 x 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 12.5 × 10⁶ 7 Individual or approximately 12.5 x 10 7 From individual CD3+CAR-expressing cells, 13.5 × 10⁶ 7 Individual or approximately 13.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 13.5 × 10⁶ 7 Individual or approximately 13.5 × 10 7 From individual CD3+CAR-expressing cells, 14.5 × 10⁶ 7 Individual or approximately 14.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 14.5 × 10⁶ 7 Individual or approximately 14.5 × 10 7 From individual CD3+CAR-expressing cells, 15.5 × 10⁶ 7 pc or approximately 15.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 15.5 × 10⁶ 7 pc or approximately 15.5 × 10 7 From individual CD3+CAR-expressing cells, 16.5 × 10⁶ 7 pc or approximately 16.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 16.5 × 10⁶ 7 pc or approximately 16.5 × 10 7From individual CD3+CAR-expressing cells, 17.5 × 10⁶ 7 pc or approximately 17.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 17.5 × 10⁶ 7 pc or approximately 17.5 × 10 7 From individual CD3+CAR-expressing cells, 18.5 × 10⁶ 7 pc or approximately 18.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 18.5 × 10⁶ 7 pc or approximately 18.5 × 10 7 From individual CD3+CAR-expressing cells, 19.5 × 10⁶ 7 Individual or approximately 19.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 19.5 × 10⁶ 7 Individual or approximately 19.5 × 10 7 From individual CD3+CAR-expressing cells, 20.5 × 10⁶ 7 pc or approximately 20.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 20.5 × 10⁶ 7 pc or approximately 20.5 × 10 7 From individual CD3+CAR-expressing cells, 21.5 × 10⁶ 7 Individual or approximately 21.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 21.5 × 10⁶ 7 Individual or approximately 21.5 × 10 7 From individual CD3+CAR-expressing cells, 22.5 × 10⁶ 7 Individual or approximately 22.5 × 10 7 These are individual CD3+CAR-expressing cells. In some embodiments, the dose is 22.5 × 10⁶ 7 Individual or approximately 22.5 × 10 7 From individual CD3+CAR-expressing cells, 23.5 × 10⁶ 7 Individual or approximately 23.5 × 10 7 These are individual CD3+CAR expressing cells.

[0158] In some embodiments, the dose is 1.0 × 10 7 individual or approximately 1.0 x 107 10⁴ CD3+CAR-expressing cells, 2.0 × 10⁶ 7 1 or approximately 2.0 x 10 7 Individual CD3+CAR-expressing cells, 3.0 × 10⁶ 7 1 or approximately 3.0 x 10 7 Individual CD3+CAR-expressing cells, 4.0 × 10⁶ 7 Individual or approximately 4.0 x 10 7 Individual CD3+CAR-expressing cells, 5.0 × 10⁶ 7 1 or approximately 5.0 x 10 7 Individual CD3+CAR-expressing cells, 6.0 × 10⁶ 7 Individual or approximately 6.0 x 10 7 Individual CD3+CAR-expressing cells, 7.0 × 10⁶ 7 1 or approximately 7.0 x 10 7 Individual CD3+CAR-expressing cells, 8.0 × 10⁶ 7 1 or approximately 8.0 x 10 7 Individual CD3+CAR-expressing cells, 9.0 × 10⁶ 7 1 or approximately 9.0 x 10 7 Individual CD3+CAR-expressing cells, 10.0 × 10⁶ 7 individual or approximately 10.0 x 10 7 Individual CD3+CAR-expressing cells, 11.0 × 10⁶ 7 Individual or approximately 11.0 x 10 7 Individual CD3+CAR-expressing cells, 12.0 × 10⁶ 7 Individual or approximately 12.0 x 10 7 Individual CD3+CAR-expressing cells, 13.0 × 10⁶ 7 Individual or approximately 13.0 x 10 7 Individual CD3+CAR-expressing cells, 14.0 × 10⁶ 7 Individual or approximately 14.0 x 10 7 Individual CD3+CAR-expressing cells, 15.0 × 10⁶ 7 individual or approximately 15.0 x 10 7 Individual CD3+CAR-expressing cells, 16.0 × 10⁶ 7 Individual or approximately 16.0 x 10 7 Individual CD3+CAR-expressing cells, 17.0 × 10⁶ 7 Individual or approximately 17.0 x 10 7 Individual CD3+CAR-expressing cells, 18.0 × 10⁶ 7Individual or approximately 18.0 x 10 7 Individual CD3+CAR-expressing cells, 19.0 × 10⁶ 7 Individual or approximately 19.0 x 10 7 Individual CD3+CAR-expressing cells, 20.0 × 10⁶ 7 individual or approximately 20.0 x 10 7 Individual CD3+CAR-expressing cells, 21.0 × 10⁶ 7 individual or approximately 21.0 × 10 7 Individual CD3+CAR-expressing cells, 22.0 × 10⁶ 7 Individual or approximately 22.0 x 10 7 Individual CD3+CAR-expressing cells, 23.0 × 10⁴ 7 Individual or approximately 23.0 x 10 7 A single CD3+CAR-expressing cell, or 24.0 × 10⁶ cells. 7 Individual or approximately 24.0 x 10 7 These are individual CD3+CAR expressing cells.

[0159] In some embodiments, the dose is 1.0 × 10 7 individual or approximately 1.0 × 10 7 Contains 10 CD3+CAR-expressing cells. In some embodiments, the dose is 2.0 × 10⁶ 7 1 or approximately 2.0 × 10 7 Contains 10 CD3+CAR-expressing cells. In some embodiments, the dose is 3.0 × 10⁶ 7 pc or approximately 3.0 × 10 7 Contains 10 CD3+CAR-expressing cells. In some embodiments, the dose is 4.0 × 10⁶ 7 pc or approximately 4.0 × 10 7 Contains 10 CD3+CAR-expressing cells. In some embodiments, the dose is 6.0 × 10⁶ 7 pc or approximately 6.0 × 10 7 Contains 10 CD3+CAR-expressing cells. In some embodiments, the dose is 8.0 × 10⁶ 8 pc or approximately 8.0 × 10 8 Contains individual CD3+CAR-expressing cells.

[0160] In some embodiments, the dose is 8.0 × 10 7 pc or approximately 8.0 × 10 7Contains 10 CD3+CAR-expressing cells. In some embodiments, the dose is 16.0 × 10⁶ 7 Individual or approximately 16.0 × 10 7 Contains individual CD3+CAR-expressing cells.

[0161] In some embodiments, the dose is 1.0 × 10 7 individual or approximately 1.0 × 10 7 Contains 10 CD4+ / CD8+CAR-expressing cells. In some embodiments, the dose is 2.0 × 10⁶ 7 1 or approximately 2.0 × 10 7 Contains 10 CD4+ / CD8+CAR-expressing cells. In some embodiments, the dose is 3.0 × 10⁶ 7 pc or approximately 3.0 × 10 7 Contains 10 CD4+ / CD8+CAR-expressing cells. In some embodiments, the dose is 4.0 × 10⁶ 7 pc or approximately 4.0 × 10 7 Contains 10 CD4+ / CD8+CAR-expressing cells. In some embodiments, the dose is 6.0 × 10⁶ 7 pc or approximately 6.0 × 10 7 Contains 10 CD4+ / CD8+CAR-expressing cells. In some embodiments, the dose is 8.0 × 10⁶ 8 pc or approximately 8.0 × 10 8 Contains individual CD4+ / CD8+CAR-expressing cells.

[0162] In some embodiments, the dose is 8.0 × 10 7 pc or approximately 8.0 × 10 7 Contains 10 CD4+ / CD8+CAR-expressing cells. In some embodiments, the dose is 16.0 × 10⁶ 7 Individual or approximately 16.0 × 10 7 Contains individual CD4+ / CD8+CAR-expressing cells.

[0163] In some embodiments, the dose of cells, for example recombinant receptor-expressing T cells, is administered to the subject as a single dose or only once within a period of 2 weeks, 1 month, 3 months, 6 months, 1 year, or longer. In some embodiments, the patient is administered multiple doses, and each dose or the total dose may be within any of the above values.

[0164] In some embodiments, engineered cells or compositions of engineered cells for administration exhibit properties that indicate or are consistent with cellular health. In some embodiments, 70, 75, 80, 85, or 90% or about 70, 75, 80, 85, or 90% or at least 70, 75, 80, 85, or 90% or at least about 70, 75, 80, 85, or 90% of such doses of CAR+ cells exhibit one or more properties or phenotypes that indicate cellular health or biologically active CAR cells, such as the absence of expression of apoptotic markers.

[0165] In certain embodiments, the phenotype is the absence of, or includes, the absence of, signs of apoptosis and / or the cell undergoing the apoptotic process. Apoptosis is a programmed cell death process that involves a set of typical morphological and biochemical events leading to characteristic cellular changes and death, including vesicle formation, cell shrinkage, nuclear fragmentation, chromatin condensation, chromosomal DNA fragmentation, and global mRNA degradation. In some aspects, the early stages of apoptosis may be indicated by the activation of certain caspases, e.g., 2, 8, 9, and 10. In some aspects, the mid-to-late stages of apoptosis are characterized by further loss of membrane integrity, chromatin condensation, and DNA fragmentation, which include biochemical events such as the activation of caspases 3, 6, and 7.

[0166] In certain embodiments, the phenotype is negative expression of one or more factors associated with programmed cell death, such as pro-apoptotics known to initiate apoptosis, e.g., members of the cell death receptor pathway, activated members of the mitochondrial (endogenous) pathway, e.g., Bcl-2 family members, e.g., Bax, Bad, and Bid, and caspases. In certain embodiments, the phenotype is the absence of indicators, e.g., annexin V molecules or TUNEL staining, that preferentially bind to apoptotic cells when incubated with or in contact with a cellular composition. In some embodiments, the phenotype is the expression of one or more markers indicating an apoptotic state in cells, or includes them. In some embodiments, the phenotype is the absence of caspase expression and / or activation, e.g., caspase-3. In some aspects, caspase-3 activation indicates increased or restarted apoptosis. In certain embodiments, caspase activation can be detected by known methods. In some embodiments, antibodies that specifically bind to activated caspases (i.e., specifically to cleaved polypeptides) may be used to detect caspase activation. In certain embodiments, the phenotype is or includes active caspase 3-. In some embodiments, the marker of apoptosis is a reagent that detects features in cells associated with apoptosis. In certain embodiments, the reagent is the annexin V molecule.

[0167] In some embodiments, a composition containing engineered cells for administration contains a certain number or quantity of cells exhibiting a phenotype indicating or consistent with cellular health. In some embodiments, less than 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the CAR-expressing T cells in a dose of engineered T cells express an apoptosis marker, 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 a dose of engineered T cells express annexin V or active caspase 3.

[0168] In some embodiments, cells, binding molecules, or recombinant receptors are administered as part of a combination treatment, simultaneously with, for example, another therapeutic intervention, such as another antibody or engineered cells or receptors, or a drug, such as a cytotoxic agent or therapeutic agent, or sequentially in any order.

[0169] Cells, binding molecules, and / or recombinant receptors are administered in combination, in some embodiments, with one or more additional therapeutic agents or in conjunction with another therapeutic intervention, either concurrently or sequentially in any order. In some situations, cells are administered in combination with another therapy in sufficiently close temporal proximity, resulting in the cell population enhancing the effect of one or more additional therapeutic agents, or vice versa. In some embodiments, cells, binding molecules, and / or recombinant receptors are administered before one or more additional therapeutic agents. In some embodiments, cells, binding molecules, and / or recombinant receptors are administered after one or more additional therapeutic agents.

[0170] B. Efficacy, efficacy, and survival 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 assessing the disease state. Exemplary methods for assessing the disease state include measurement of M protein in biological fluids, e.g., blood and / or urine, by electrophoresis and immunofixation, quantification of sFLC(κ and λ) in blood, skeletal examination, and positron emission tomography (PET) / computed tomography (CT) imaging in subjects with extramedullary disease. In some embodiments, the disease state may be assessed by bone marrow examination. In some examples, the dose and / or frequency of administration is determined by the expansion and persistence of recombinant receptors or cells in the blood and / or bone marrow. In some embodiments, the dose and / or frequency of administration is determined based on the antitumor activity of recombinant receptors or engineered cells. In some embodiments, antitumor activity is determined by the overall response rate (ORR) and / or the International Myeloma Working Group (IMWG) Uniform Response Criteria (see Kumar et al. (2016) Lancet Oncol 17(8):e328-346). In some embodiments, response is assessed using minimal residual disease (MRD) assessment. In some embodiments, MRD may be assessed by methods such as flow cytometry and high-throughput sequencing, e.g., deep sequencing. In some aspects, subjects with MRD-negative disease are 10 5 1 nucleated cell in the middle or higher minimum sensitivity (i.e., 10 -5 Examples include subjects exhibiting the absence of abnormal clonal plasma cells in bone marrow aspirate, which can be excluded by assays with high sensitivity, such as flow cytometry (next-generation flow cytometry; NGF) or high-throughput sequencing, such as deep sequencing or next-generation sequencing (NGS).

[0171] In some contexts, sustained MRD negativity includes subjects exhibiting MRD negativity in the spinal cord (NGF or NGS, or both) and by imaging as defined below, confirmed at intervals of at least one year. Subsequent evaluation may be used to further identify the duration of negativity (e.g., MRD negativity at 5 years). In some contexts, flow MRD negativity is 10 5 This includes subjects exhibiting the absence of phenotypically abnormal clonal plasma cells by NGF in bone marrow aspirate using the EuroFlow standard operating procedure (or validated equivalent method) for MRD detection in multiple myeloma with higher minimum sensitivity or nucleated cells. In some aspects, sequencing MRD negativity indicates the presence of clones, 10 5 This includes subjects exhibiting the absence of clonal plasma cells by NGS in bone marrow aspirate, defined as fewer than two identical sequencing reads obtained after DNA sequencing of bone marrow aspirate using one nucleated cell or a LymphoSIGHT platform (or validated equivalent method) with higher minimum sensitivity. In some aspects, imaging plus MRD negativity includes subjects exhibiting MRD negativity, assessed by the disappearance of any compartment of increased tracer uptake found in baseline or preceding PET / CT, or a reduction to a lower mediastinal blood pool SUV or a reduction to less than normal surrounding tissue, in addition to NGF or NGS (see Kumar et al. (2016) Lancet Oncol 17(8):e328-346).

[0172] In some embodiments, response is assessed based on the duration of response after administration of recombinant receptors or cells. In some examples, the dose and / or frequency of administration may be based on toxicity. In some embodiments, the dose and / or frequency may be determined based on the health-related quality of life (HRQoL) of the subject to whom the recombinant receptors and / or cells are administered. In some embodiments, the dose and / or frequency of administration may be modified, i.e., increased or decreased, based on any of the above criteria.

[0173] In some aspects, the survival of the subject, survival within a specific period, degree of survival, presence or duration of event-free or asymptomatic survival, or recurrence-free survival are evaluated. In some embodiments, any symptoms of the disease or condition are evaluated. In some embodiments, the degree of tumor burden is identified. In some embodiments, exemplary parameters for determination include specific clinical outcomes indicating remission or improvement in the tumor. Such parameters include duration of disease control, including objective response (OR), complete response (CR), severe 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 assessed using minimal residual disease (MRD) assessment. Specific thresholds for parameters may be set to determine the effectiveness of the methods provided herein. In some embodiments, the disease or disorder treated is multiple myeloma. In some embodiments, measurable disease criteria for multiple myeloma may include (1) serum M protein of 1 g / dL or higher; (2) urinary M protein of 200 mg or higher per 24 hours; and (3) serum free light chain (sFLC) levels of 10 mg / dL or higher with an abnormal κ-to-λ ratio. In some cases, light chain disease is acceptable only for subjects who do not have measurable disease in serum or urine.

[0174] In some aspects, for example, depending on the embodiment provided, the response to the therapy may be measured at a specified time after the initiation of administration of the cell therapy. In some embodiments, the specified time is within the range defined by 1, 2, 3, 6, 9, 12, 18, 24, 30, or 36 months after the initiation of administration, or approximately 1, 2, 3, 6, 9, 12, 18, 24, 30, or 36 months after the initiation of administration, or any of the above. In some embodiments, the specified time is within the range defined by 4, 8, 12, 16, 20, 24, 28, 32, 36, 48, or 52 weeks after the initiation of administration, or any of the above. In some embodiments, the specified time is 1 month after the initiation of administration or approximately 1 month after. In some embodiments, the specified time is 3 months after the initiation of administration or approximately 3 months after. In some embodiments, the specified time is 6 months after the initiation of administration or approximately 6 months after. In some embodiments, the specified time is 9 months after the initiation of administration or approximately 9 months after. In some embodiments, the specified time is 12 months or approximately 12 months after the start of administration.

[0175] In some embodiments, the response or outcome determined 3, 6, 9, or 12 months after a specified time point, or approximately 3, 6, 9, or 12 months later, is equal to or improved upon the response or outcome determined at the initial specified time point. For example, in some cases, if the response or outcome determined at an initial specified time point is stable disease (SD), progressive disease (PD), or relapse, subjects treated according to the provided embodiment may, at a subsequent time point, 3, 6, 9, or 12 months after the initial specified time point, or approximately 3, 6, 9, or 12 months later, demonstrate a response or outcome equal to, or equal to, or improved to, the response or outcome at the initial specified time point, which is an objective response (OR), complete response (CR), severe complete response (sCR), best partial response (VGPR), or partial response (PR) (e.g., demonstrating a better response outcome according to the International Myeloma Working Group (IMWG) Uniform Response Criteria; see Kumar et al. (2016) Lancet Oncol 17(8):e328-346). In some cases, subjects treated according to the provided embodiment may demonstrate an improved response or outcome between the two time points of determination. In some cases, subjects may exhibit a partial response (PR) or broad-spectrum response (VGPR) at an initial designated time point for evaluation, for example, four weeks after the start of administration, and then an improved response, such as a complete response (CR) or progressive complete response (sCR), at a later time point, for example, 12 weeks after the start of administration. In some cases, progression-free survival (PFS) is described as the length of time during and after treatment for a disease, such as cancer, during which the subject survives with the disease but without disease progression. In some cases, objective response (OR) is described as a measurable response. In some cases, objective response rate (ORR; also known in some cases as overall response rate) is described as the percentage of patients who achieved a complete response (CR) or a partial response (PR).In some contexts, overall survival (OS) is described as the length of time from the date of diagnosis of a disease, such as cancer, or the date of initiation of treatment, during which a subject diagnosed with the disease is still alive. In some contexts, event-free survival (EFS) is described as the length of time after treatment for cancer resolution, during which the subject remains free from certain complications or events that the treatment is intended to prevent or delay. These events may include a recurrence of cancer, the onset of certain symptoms, such as bone pain from cancer spreading to the bone, or death.

[0176] In some embodiments, the degree of duration of response (DOR) may be defined as the time from the recording of tumor response to disease progression. In some embodiments, parameters for evaluating response may include sustained response, e.g., response that persists over a period of time from the initiation of therapy. In some embodiments, sustained response is indicated by the response rate 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, or 24 months after the initiation of therapy, or approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, or 24 months after the initiation of therapy. In some embodiments, the response or outcome persists for more than 3, 6, 9, or 12 months, or approximately 3, 6, 9, or 12 months.

[0177] In some embodiments, the Eastern Cooperative Oncology Group (ECOG) Performance Status Scale can be used to assess or select subjects for treatment, e.g., subjects who have had poor outcomes from previous 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 functionality in terms of their ability to care for themselves, their daily activity level, and their physical abilities (e.g., walking, working). In some embodiments, an ECOG Performance Status of 0 indicates that the subject is able to perform normal activities. In some cases, a subject with an ECOG Performance Status of 1 exhibits some limitations in physical activity, but is fully able to walk. In some cases, a patient with an ECOG Performance Status of 2 is able to walk more than 50% of the time. In some cases, subjects with an ECOG performance status of 2 may also be capable of caring for themselves; see, for example, Sorensen et al., (1993) Br J Cancer 67(4) 773-775. In some embodiments, subjects administered according to the methods or treatment regimens provided herein include subjects with an ECOG performance status of 0 or 1.

[0178] In some embodiments, administration by the method provided effectively treats a subject even if the subject has become resistant to another therapy. In some embodiments, when administered to a subject according to the embodiments described herein, the dose or composition has the ability to achieve an objective response (OR) in at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the administered subjects. In some embodiments, the OR includes subjects that achieve a severe complete response (sCR), a complete response (CR), a best partial response (VGPR), a partial response (PR), or a minimal response (MR). In some embodiments, when administered to a subject according to the embodiments described herein, the dose or composition has the ability to achieve a severe complete response (sCR), a complete response (CR), a best partial response (VGPR), or a partial response (PR) in at least 50%, 60%, 70%, 80%, or 85% of the administered subjects. In some embodiments, when administered to a subject according to the embodiments described herein, the dose or composition has the ability to achieve a severe complete response (sCR) or complete response (CR) in at least 20%, 30%, 40%, 50%, 60%, or 70% of the subjects administered. In some aspects, the specific response to a treatment, e.g., the method provided herein, may be evaluated based on the International Myeloma Working Group (IMWG) Uniform Response Criteria (see Kumar et al. (2016) Lancet Oncol 17(8):e328-346).

[0179] In some embodiments, administration by the provided method effectively treats subjects even if they have become resistant to other therapies. 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 subjects treated according to the method achieve objective response (OR). In some embodiments, at least 50% or at least about 50%, at least 60% or at least about 60%, at least 70% or at least about 70%, at least 80% or at least about 80%, or at least 90% or at least about 90% of subjects treated according to the method achieve CR and / or objective response (OR). In some embodiments, the criteria evaluated for effective treatment include 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).

[0180] 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 3 months, 6 months or 12 months or about 3 months, 6 months or 12 months or about 13 months or about 14 months, and on average, subjects treated according to the methods exhibit median PFS or OS of 6 months, 12 months or 18 months or about 6 months, 12 months or 18 months, and / or subjects exhibit PFS or OS after therapy for at least 6, 12, 18 months or more or at least about 6, 12, 18 months or more or longer.

[0181] In some embodiments, subjects treated according to the provided method exhibit 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 having the best overall response (BOR) up to 12 months, 18 months, 24 months, 36 months, or longer.

[0182] C. Toxicity In some embodiments, the methods provided include, for example, alternative cell therapies, such as alternative CARs. + The administration of T cell compositions and / or alternative dosing of cells, for example, compared to dosing of cells not administered in a defined ratio, is designed to include or includes features that result in lower rates and / or lower degrees of toxicity, toxicity outcomes or symptoms, toxicity-promoting profiles, factors, or characteristics, such as cytokine release syndrome (CRS) or neurotoxicity (NT). Cytokine release syndrome (CRS) and neurotoxicity may be graded according to the American Society for Transplantation and Cellular Therapy (ASTCT) Consensus Grading System (see, for example, Lee et al. Biol Blood Marrow Transplant. 2019 Apr;25(4):625-38).

[0183] In some aspects, the lower differentiation state of the manipulated T cells administered as part of the method provided herein (e.g., a higher percentage of naive or central memory phenotypes, e.g., CCR7) + CD45RA + CD27 + CCR7 + , or CD62L - CCR7 +While engineered T cells (having a selected phenotype) are expected to be more active than more differentiated cells, the findings indicate that the safety of cell therapy can be successfully managed. In some aspects, providing lower doses of the composition compared to cell compositions produced by methods involving more differentiated cells, such as methods involving cell expansion and proliferation, achieves robust efficacy and high safety. In some aspects, it has been found that higher doses of cells of the provided anti-BCMA CAR composition can be administered while maintaining a lower degree of toxicity, such as severe cytokine release syndrome (CRS) or severe neurotoxicity. Therefore, in some aspects, the provided method can be used to provide alternative cell therapies, such as alternative CARs containing more differentiated engineered T cells than those administered in the present invention. + Compared to methods involving the administration of a T cell composition, a higher dose of engineered T cells (e.g., 50 × 10) is used. 6 More than 100 CAR-expressing T cells, for example, 100 × 10⁶ 6 1 or approximately 100 x 10 6 CAR-expressing T cells, 160 × 10⁶ 6 Individual or approximately 160 x 10 6 10 CAR-expressing T cells, or 200 × 10⁶ 6 1 or approximately 200 x 10 6 This includes the administration of individual CAR-expressing T cells.

[0184] In some embodiments, the methods provided do not result in a high rate or high probability of toxicity or toxic outcomes, in other words, they reduce the rate or probability of toxicity or toxic outcomes, such as neurotoxicity (NT) and cytokine release syndrome (CRS), compared to, for example, some other cell therapies. In some embodiments, the methods do not result in, nor increase the risk of, severe NT (sNT), severe CRS (sCRS), macrophage activation syndrome, tumor lysis syndrome, fever of at least 38°C or at least about 38°C for three or more days, and plasma CRP levels of at least 20 mg / dL or at least about 20 mg / dL. In some embodiments, 30%, 35%, 40%, 50%, 55%, 60% or more of subjects treated according to the methods provided do not exhibit any grade of CRS or any grade of neurotoxicity. In some embodiments, less than 50% of the treated subjects (e.g., at least 60%, at least 70%, at least 80%, at least 90%, or more of the treated subjects) exhibit cytokine release syndrome (CRS) of grade 2 or higher and / or neurotoxicity of grade 2 or higher. In some embodiments, at least 50% of the subjects treated according to the method (e.g., at least 60%, at least 70%, at least 80%, at least 90%, or more of the treated subjects) do not exhibit severe toxic outcomes (e.g., severe CRS or severe neurotoxicity); for example, they do not exhibit neurotoxicity of grade 3 or higher and / or severe CRS; or they do not exhibit such outcomes within a certain period after treatment, for example, within one week, two weeks, or one month after cell administration. In some embodiments, parameters evaluated to determine a particular toxicity include adverse events (AEs), dose-limiting toxicity (DLTs), CRS, and NTs.

[0185] The administration of adoptive T-cell therapy, such as treatment using T cells expressing chimeric antigen receptors, may cause toxic effects or outcomes, including cytokine release syndrome and neurotoxicity. In some cases, such effects or outcomes occur concurrently with high levels of circulating cytokines, which may underlie the observed toxicity.

[0186] In some cases, toxic outcomes are cytokine release syndrome (CRS) or severe CRS (sCRS), associated with cytokine release syndrome (CRS) or severe CRS (sCRS), or exhibit cytokine release syndrome (CRS) or severe CRS (sCRS). CRS, such as sCRS, may occur in some cases after adoptive T-cell therapy and after administration of other biological products to the subject. 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); 2709-2720 (2012); see Xu et al., Cancer Letters 343 (2014) 172-78.

[0187] Typically, CRS is caused by an aggravated systemic immune response mediated, for example, by T cells, B cells, NK cells, monocytes, and / or macrophages. Such cells may release large amounts of inflammatory mediators, such as cytokines and chemokines. Cytokines may trigger an acute inflammatory response and / or induce endothelial organ damage that can lead to 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 may include cardiotoxicity, respiratory distress, neurotoxicity, and / or hepatic failure. In some aspects, fever, particularly high fever (≥38.5°C or ≥101.3°F), is associated with CRS or its risk. In some cases, the features or symptoms of CRS mimic infections. In some aspects, infections are also considered in subjects exhibiting CRS symptoms, and culture-based monitoring and empirical antibiotic therapy may be administered. Other symptoms associated with CRS may include cardiac dysfunction, adult respiratory distress syndrome, renal and / or hepatic failure, coagulation disorders, disseminated intravascular coagulation, and capillary leak syndrome.

[0188] CRS may be treated with anti-inflammatory therapies, such as anti-IL-6 therapies, such as anti-IL-6 antibodies, such as tocilizumab, or antibiotics, or other agents as described herein. The outcomes, signs, and symptoms of CRS are publicly known and include those described herein. In some embodiments, if a particular regimen or administration results in a particular CRS-related outcome, sign, or symptom, or if a particular regimen or administration does not result in a particular CRS-related outcome, sign, or symptom, then a particular outcome, sign, and symptom, and / or its quantity or degree, may be specified.

[0189] In the context of CAR-expressing cell administration, CRS typically occurs 6–20 days after infusion of CAR-expressing cells. 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 loading at the time of infusion. Generally, CRS is accompanied by elevated serum levels of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, and / or interleukin (IL)-2. Other cytokines that can be rapidly induced in CRS are IL-1β, IL-6, IL-8, and IL-10.

[0190] Exemplary CRS-related outcomes include fever, rigidity, chills, hypotension, dyspnea, acute respiratory distress syndrome (ARDS), encephalopathy, elevated ALT / AST, renal failure, cardiac events, hypoxia, neuropathy, and death. Neurological complications include delirium, seizure-like activity, confusion, word-finding difficulty, aphasia, and / or stupor. Other CRS-related outcomes include fatigue, nausea, headache, seizures, tachycardia, myalgia, rash, acute vasoleap syndrome, impaired hepatic function, and renal failure. In some aspects, CRS is associated with one or more factors, such as elevated serum ferritin, d-dimer, aminotransferase, lactate dehydrogenase, and triglycerides, or with hypofibrinogenemia or hepatosplenomegaly. Other exemplary signs or symptoms associated with CRS include hemodynamic instability, febrile neutropenia, elevated serum C-reactive protein (CRP), altered coagulation parameters (e.g., international normalized ratio (INR), prothrombin time (PTI), and / or fibrinogen), altered cardiac and other organ function, and / or absolute neutrophil count (ANC).

[0191] In some embodiments, CRS-related outcomes include: persistent fever, e.g., fever for more than 2 days, e.g., more than 3 days, e.g., more than 4 days, or for at least 3 consecutive days, at a specified temperature, e.g., fever above 38°C or about 38°C; fever above 38°C or about 38°C; elevated cytokine levels, e.g., maximum multiplier change of at least two cytokines (e.g., at least two from the group consisting of interferon-gamma (IFNγ), GM-CSF, IL-6, IL-10, Flt-3L, fractalkine, and IL-5, and / or tumor necrosis factor α (TNFα)) compared to pre-treatment levels, e.g., maximum multiplier change of at least 75 or at least about 75, or maximum multiplier change of at least one such cytokine, e.g., maximum multiplier change of at least 250 or at least about 250; and / or at least one clinical sign of toxicity, e.g., hypotension (e.g., at least one intravenous vasoactive vasopressor). This includes hypoxia (e.g., plasma oxygen (PO2) levels below 90% or approximately below 90%) as measured by a pressor; and / or one or more neurological disorders (including altered mental state, numbness, and seizures). In some embodiments, neurotoxicity (NT) may be observed concurrently with CRS.

[0192] Exemplary CRS-related outcomes include elevated or high serum levels of one or more factors, including cytokines and chemokines, as well as other factors associated with CRS. Furthermore, exemplary outcomes include increased synthesis or secretion of one or more such factors. Such synthesis or secretion may be mediated by T cells, or cells interacting with T cells, such as innate immune cells or B cells.

[0193] In some embodiments, CRS-related serofactors or CRS-related outcomes include inflammatory cytokines and / or chemokines, including interferon-γ (IFN-γ), TNF-α, 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 α (TNFα), IL-6, and IL-10, IL-1β, IL-8, IL-2, MIP-1, Flt-3L, fractalkines, and / or IL-5. In some embodiments, the factors or outcomes include C-reactive protein (CRP). CRP is an early and readily measurable risk factor for CRS, as well as a cell proliferation marker. In some embodiments, subjects who are measured to have high levels of CRP, e.g., ≥15 mg / dL, have CRS. In some embodiments, subjects measured to have high levels of CRP do not have CRS. In some embodiments, the measurement of CRS includes the measurement of CRP and another factor indicating CRS.

[0194] 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.

[0195] To predict which patients are likely to develop sCRS, CRS decision criteria have been developed that appear to correlate with the development of CRS (see Davilla et al. Science translational medicine. 2014;6(224):224ra25). Factors include fever, hypoxia, hypotension, neurological changes, and elevated serum levels of seven cytokines (IFNγ, IL-5, IL-6, IL-10, Flt-3L, fractalkine, and GM-CSF), whose elevated levels induced by treatment can correlate well with pre-treatment tumor burden and sCRS symptoms. Other guidelines for the diagnosis and management of CRS are publicly available (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 cases, the criteria for determining the CRS grade are those detailed in Table 2 below.

[0196] (Table 2) Exemplary Grade Classification Criteria for CRS TIFF2026113544000005.tif129164

[0197] In some aspects, the criteria reflecting the CRS grade are detailed in Table 3 below.

[0198] (Table 3) Exemplary Grade Classification Criteria for CRS TIFF2026113544000006.tif74165

[0199] In some embodiments, high-dose vasopressor therapy includes those listed in Table 4 below.

[0200] (Table 4) High-dose vasopressors (all doses require at least 3 hours of administration) TIFF2026113544000007.tif42165 aVASST test vasopressor equivalent formula: Norepinephrine equivalent = [Norepinephrine (μg / min)] + [Dopamine (μg / kg / min) ÷ 2] + [Epinephrine (μg / min)] + [Phenylephrine (μg / min) ÷ 10]

[0201] In some embodiments, the toxicity outcome is severe CRS. In some embodiments, the toxicity outcome is the absence of severe CRS (e.g., moderate or mild CRS). In some embodiments, after administration, the subject (1) has a fever of at least 38°C for at least 3 days, (2) (a) a maximum multiplier change of at least 75 for at least two of the following seven cytokines: interferon-gamma (IFNγ), GM-CSF, IL-6, IL-10, Flt-3L, fracktalkine, and IL-5 compared to the level immediately after administration, and / or (b) a maximum multiplier change of at least 75 for at least two of the following seven cytokines: interferon-gamma (IFNγ), GM-CSF, IL-6 A patient is considered to have developed “severe CRS” (“sCRS”) in response to or secondarily to the administration of a dose of cell therapy or its cells if (c) an elevation of cytokines including a maximum multiplier change of at least 250 for at least one of the groups IL-10, Flt-3L, fractalkine, and IL-5, and (c) at least one clinical sign of toxicity, such as hypotension (requiring at least one intravenous vasopressor) or hypoxia (PO2 < 90%) or one or more neurological disorders (including altered mental status, blunting, and / or seizures). In some embodiments, severe CRS includes CRS having a grade of 3 or higher, such as those listed in Tables 2 and 3.

[0202] In some embodiments, toxic outcomes, such as the level of CRS-related outcomes, such as serum levels of CRS indicators, are measured by ELISA. In some embodiments, fever and / or C-reactive protein (CRP) levels may be measured. In some embodiments, subjects with fever and CRP ≥ 15 mg / dL may be considered at high risk for developing severe CRS. In some embodiments, CRS-related serofactors or CRS-related outcomes include increases in the levels and / or concentrations of inflammatory cytokines and / or chemokines, including Flt-3L, fractalkines, 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 readily measurable risk factor for CRS, CRP is also a marker of cell proliferation. In some embodiments, subjects who are measured to have high levels of CRP, such as ≥15 mg / dL, have CRS. In some embodiments, subjects who are measured to have high levels of CRP do not have CRS. In some embodiments, the degree of CRS includes the degree of CRP and another factor that indicates CRS.

[0203] In some embodiments, outcomes associated with severe CRS or Grade 3 or higher CRS, e.g., Grade 4 or higher, include one or more of the following: persistent fever, e.g., for two or more days, e.g., three or more days, e.g., four or more days, or at least three consecutive days, at a specified temperature, e.g., above 38°C or approximately above 38°C; fever, e.g., above 38°C or approximately above 38°C; increased cytokines, e.g., at least two cytokines (e.g., interferon-gamma (IFNγ), GM-CSF, IL-6, IL-10, Flt-3L, fractalkine, and IL-5), and / or tumor necrosis. A maximum percentage change of, for example, at least 75 or at least about 75 compared to pre-treatment levels of at least two cytokines belonging to the group consisting of factor α (TNFα), or a maximum percentage change of, for example, at least 250 or at least about 250 for at least one of such cytokines; and / or at least one clinical sign of toxicity, e.g., hypotension (e.g., measured using at least one intravenous vasopressor); hypoxia (e.g., plasma oxygen (PO2) levels less than 90% or about 90%); and / or one or more neurological disorders (including altered mental status, blunting, and seizures). In some embodiments, severe CRS includes CRS requiring management or nursing in an intensive care unit (ICU).

[0204] In some embodiments, CRS, e.g., severe CRS, comprises a combination of (1) persistent fever (fever of at least 38°C for at least 3 days) and (2) a serum CRP level of at least 20 mg / dL or at least about 20 mg / dL. In some embodiments, CRS comprises hypotension requiring two or more vasopressors or respiratory failure requiring mechanical ventilation. In some embodiments, the dose of the vasopressor is increased in the second or subsequent administration.

[0205] In some embodiments, severe CRS or Grade 3 CRS includes increased alanine aminotransferase, increased aspartate aminotransferase, chills, febrile neutropenia, headache, left ventricular dysfunction, encephalopathy, hydrocephalus, and / or tremor.

[0206] You can specify methods for measuring or detecting various outcomes.

[0207] In some aspects, toxic outcomes are neurotoxic or related to neurotoxicity. In some embodiments, symptoms associated with the clinical risk of neurotoxicity include confusion, delirium, aphasia, expressive aphasia, hypotonia, myoclonus, lethargy, altered mental status, convulsions, seizure-like activity, seizures (optionally confirmed by electroencephalography (EEG)), high levels of β-amyloid (Aβ), high levels of glutamate, and high 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)).

[0208] In some cases, neurological symptoms may be among the earliest symptoms of sCRS. In some embodiments, neurological symptoms may begin 5–7 days after cell therapy infusion. In some embodiments, the duration of neurological changes may be 3–19 days. In some cases, recovery of neurological changes occurs after the resolution of other symptoms of sCRS. In some embodiments, the time or extent of resolution of neurological changes is not accelerated by treatment with anti-IL-6 and / or steroids.

[0209] In some embodiments, a subject is considered to have developed “severe neurotoxicity” in response to or in association with the administration of cell therapy or a dose of cells if, after administration, the subject exhibits symptoms that limit self-care (e.g., bathing, dressing, eating, using the toilet, taking medication), including: 1) symptoms of peripheral motor neuropathy, including inflammation or degeneration of peripheral motor nerves; 2) symptoms of peripheral sensory neuropathy, including inflammation or degeneration of peripheral sensory nerves; dysesthesia, e.g., distortion of sensory perception causing abnormal and unpleasant sensations; neuralgia, e.g., severe pain along a nerve or group of nerves; and / or parestesia, e.g., dysfunction of sensory neurons causing abnormal skin sensations of stinging, numbness, pressure, cold, and warmth in the absence of stimulation. In some embodiments, severe neurotoxicity includes neurotoxicity of grade 3 or higher, e.g., the neurotoxicities shown in Table 5.

[0210] (Table 5) Exemplary Grade Classification Criteria for Neurotoxicity TIFF2026113544000008.tif72164

[0211] In some embodiments, the method reduces symptoms associated with CRS or neurotoxicity compared to other methods. In some aspects, the method provided reduces symptoms, outcomes, or factors associated with CRS, including symptoms, outcomes, or factors associated with severe CRS or CRS of grade 3 or higher, compared to other methods. For example, subjects treated according to the method of the present invention may not have, and / or may have reduced, detectable symptoms, outcomes, or factors for CRS such as those described, for example, in Tables 2 and 3, e.g., severe CRS or CRS of grade 3 or higher. In some embodiments, subjects treated according to the method of the present invention may have reduced neurotoxic symptoms compared to subjects treated by other methods, such as, for example, limb weakness or paralysis; impaired memory, vision, and / or cognitive function; uncontrollable obsessive-compulsive and / or compulsive behavior; delusions; headaches; cognitive and behavioral problems including impaired motor control, cognitive decline, and autonomic nervous system dysfunction; and sexual dysfunction. In some embodiments, subjects treated according to the methods of the present invention may experience a reduction in symptoms associated with peripheral motor neuropathy, peripheral sensory neuropathy, paresthesia, neuralgia, or paresthesia.

[0212] In some embodiments, the method reduces neurotoxicity-related outcomes, including damage to the nervous system and / or brain, such as neuronal death. In some aspects, the method reduces levels of neurotoxicity-related factors, such as β-amyloid (Aβ), glutamate, and oxygen radicals.

[0213] In some aspects, the toxic outcome is dose-limiting toxicity (DLT). In some aspects, the toxic outcome is dose-limiting toxicity. In some aspects, the toxic outcome is the absence of dose-limiting toxicity. In some aspects, dose-limiting toxicity (DLT) is defined as any toxicity of grade 3 or higher when assessed by any known or published guidelines for assessing a particular toxicity. Such guidelines include, for example, any of the above, and the National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) version 4.0.

[0214] In some embodiments, the rate, risk, or likelihood of developing toxicity, such as CRS or neurotoxicity, or severe CRS or neurotoxicity, such as grade 3 or higher CRS or neurotoxicity, observed when a certain dose of T cells is administered is low, making it possible to administer cell therapy agents on an outpatient basis according to the provided method and / or by the provided manufactured article or composition. In some embodiments, cell therapy agents, such as T cells (e.g., CAR), can be administered according to the provided method and / or by the provided manufactured article or composition. + The administration of the relevant dose of T cells is performed on an outpatient basis or does not require hospitalization, such as an overnight stay.

[0215] In some cases, including subjects treated as outpatients, cell therapy agents, such as T cells (e.g., CAR cells), are used according to the provided method and / or by the provided manufactured articles or compositions. + Subjects receiving such doses of T cells will not receive any intervention to treat toxicity before or with the administration of such cell doses, unless they exhibit or exhibit signs or symptoms of toxicity, such as neurotoxicity or CRS. Exemplary agents for treating, delaying, reducing, or improving toxicity are described in Section II.

[0216] In some embodiments, cell therapy agents, such as T cells (e.g., CAR), include subjects treated as outpatients.+ If a subject receiving a certain dose of T cells develops a fever, the subject is given or instructed to take or administer a therapeutic agent to reduce the fever. In some embodiments, the subject's fever is characterized by a specific threshold temperature or threshold level or higher (or measured as such) of the subject's body temperature. In some aspects, the threshold temperature is associated with at least a low fever, at least a moderate fever, and / or at least a high fever. In some embodiments, the threshold temperature is a specific temperature or range. For example, the threshold temperature may be 38, 39, 40, 41, or 42°C or about 38, 39, 40, 41, or 42°C, or at least 38, 39, 40, 41, or 42°C or at least about 38, 39, 40, 41, or 42°C, and / or in the range from 38°C or about 38°C to 39°C or about 39°C, from 39°C or about 39°C to 40°C or about 40°C, from 40°C or about 40°C to 41°C or about 41°C, or from 41°C or about 41°C to 42°C or about 42°C.

[0217] In some embodiments, a treatment designed to reduce fever includes treatment using an antipyretic. The antipyretic may include any active ingredient that reduces fever, e.g., a compound, composition, or component, e.g., one of any number of active ingredients known to have an antipyretic effect, e.g., NSAIDs (such as ibuprofen, naproxen, ketoprofen, and nimeslide), salicylic acids (e.g., aspirin, choline salicylate, magnesium salicylate, and sodium salicylate), paracetamol, acetaminophen, metamizole, nabumetone, phenaxone, antipyrine, or any other antipyretic. In some embodiments, the antipyretic is acetaminophen. In some embodiments, acetaminophen may be administered orally or intravenously at a dose of 12.5 mg / kg, up to every 4 hours. In some embodiments, the antipyretic is ibuprofen or aspirin, or includes ibuprofen or aspirin.

[0218] In some embodiments, if the fever is persistent, the subject is administered an alternative treatment to address the toxicity, such as one of those described in Section II below. For subjects treated on an outpatient basis, if they have and / or are determined to have a persistent fever, they are instructed to return to the hospital. In some embodiments, a subject has and / or is determined to have or is considered to have a persistent fever if: after a designated treatment, such as treatment with an antipyretic, e.g., an NSAID or salicylic acid, e.g., ibuprofen, acetaminophen, or aspirin, the subject exhibits a fever above a reasonable threshold temperature, and the subject's fever or body temperature does not decrease or does not decrease by a specified amount or more (e.g., not by more than 1°C; generally, not by more than about 0.5°C, 0.4°C, 0.3°C, or 0.2°C, or not fluctuating by more than about 0.5°C, 0.4°C, 0.3°C, or 0.2°C). For example, a subject is considered to have a persistent fever if he or she is found to have a fever of at least 38 or 39°C or at least about 38 or 39°C, and this fever does not decrease by 0.5°C, 0.4°C, 0.3°C, or 0.2°C, or about 0.5°C, 0.4°C, 0.3°C, or 0.2°C, for 6 hours, 8 hours, 12 hours, or 24 hours, even after treatment with an antipyretic such as acetaminophen; or by more than 0.5°C, 0.4°C, 0.3°C, or 0.2°C, or by more than about 0.5°C, 0.4°C, 0.3°C, or 0.2°C, or by 1%, 2%, 3%, 4%, or 5%, or by about 1%, 2%, 3%, 4%, or 5%. In some embodiments, the dosage of the antipyretic is a dosage that is normally effective when the subject is fever or a particular type of fever, such as fever associated with a bacterial or viral infection, such as a localized or systemic infection.

[0219] In some embodiments, a subject is determined to have and / or be deemed to have a persistent fever if the subject exhibits a fever above a reasonable threshold temperature, and the fever or body temperature of the subject does not fluctuate by more than about 1°C or about 1°C, and generally by more than about 0.5°C, 0.4°C, 0.3°C, or 0.2°C, or about 0.5°C, 0.4°C, 0.3°C, or 0.2°C. Typically, such fluctuation above a certain amount or lack of fluctuation above a certain amount is measured over a predetermined period of time (e.g., over a period of 24 hours, 12 hours, 8 hours, 6 hours, 3 hours, or 1 hour; may be measured from the first sign of fever or from the first time the body temperature exceeds a specified threshold). For example, in some embodiments, a subject is considered to have a persistent fever if it exhibits a fever of at least 38 or 39°C or at least about 38 or 39°C or at least about 38 or 39°C, and its body temperature does not fluctuate by more than 0.5°C, 0.4°C, 0.3°C, or 0.2°C, or by more than about 0.5°C, 0.4°C, 0.3°C, or 0.2°C, over a period of 6 hours, 8 hours, 12 hours, or 24 hours.

[0220] In some embodiments, the fever is a persistent fever; in some aspects, at the point in time when the subject is determined to have a persistent fever, for example, from such determination, or from initial therapies that may induce toxicity, such as cell therapy, for example, CAR + The procedure is performed within 1, 2, 3, 4, 5, 6, or less of the time elapsed from the first such determination following the administration of T cells, such as T cells.

[0221] In some embodiments, one or more interventions or agents for treating toxicity, such as a toxicity-targeting therapeutic agent, are administered at or immediately after the time when the subject is determined or confirmed (e.g., initially determined or confirmed) to be exhibiting persistent fever as measured, for example, according to any of the embodiments described above. In some embodiments, one or more toxicity-targeting therapeutic agents are administered within a certain period from such confirmation or determination, for example, within 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, or 8 hours thereafter.

[0222] II. Cell therapy and cell manipulation In some embodiments, the cell therapies disclosed herein (e.g., T-cell therapies) include the step of administering engineered cells that express recombinant receptors (e.g., CARs) designed to recognize and / or specifically bind to antigens associated with a disease or condition such as r / r / MM.

[0223] In some aspects of the methods and uses provided, engineered cells, such as T cells, express a chimeric receptor, such as a chimeric antigen receptor (CAR), which contains one or more domains that combine a ligand-binding domain (e.g., an antibody or antibody fragment) that provides specificity for a desired antigen (e.g., a tumor antigen) with an intracellular signaling domain.

[0224] The embodiments provided include compositions, products, compounds, methods and uses that target or are directed toward BCMA and BCMA-expressing cells and diseases. BCMA is observed to be expressed in certain diseases and conditions, such as malignant tumors or their tissues or cells, for example, in malignant plasma cells from all relapsed or newly diagnosed myeloma patients, for example, and is heterogeneously expressed, for example, and is hardly expressed in normal tissues. The embodiments provided include approaches useful in treating such diseases and conditions and / or targeting such cell types, and these include nucleic acid molecules encoding BCMA-binding receptors, including chimeric antigen receptors (CARs), and encoded receptors such as encoded CARs, and compositions and products containing them. The receptor is generally a BCMA-specific antibody (antigen-binding antibody fragment, e.g., heavy-chain variable (V)). H The antigen-binding domain may include a region, a single-domain antibody fragment, and a single-chain fragment containing scFv. Cells containing such BCMA-binding receptors, e.g., nucleic acids expressing anti-BCMA CAR and / or encoding such receptors, e.g., engineered or recombinant cells, as well as compositions and products and therapeutic doses containing such cells, are also provided. Methods for evaluating, optimizing, manufacturing and using nucleic acid sequences, e.g., nucleic acid sequences encoding recombinant BCMA-binding receptors, are also provided. Methods for manufacturing and using cells expressing or containing recombinant BCMA-binding receptors and recombinant BCMA-binding receptor-coding polynucleotides (e.g., engineered cells) or compositions containing such cells (e.g., for use in the treatment or remission of BCMA-expressing diseases and conditions) are also provided.

[0225] Adoptive cell therapy (including the administration of cells expressing chimeric receptors specific to the disease or disorder of interest, such as chimeric antigen receptors (CARs) and other recombinant antigen receptors, as well as adoptive immunotherapy and adoptive T-cell therapy) may be effective in treating cancer and other diseases and disorders. In certain circumstances, the available adoptive cell therapy approaches may not always be entirely satisfactory. In some aspects, optimal efficacy may depend on the ability of administered cells to recognize and bind to targets, such as target antigens like BCMA; to be transported to appropriate sites within the target, tumor, and its environment, to localize, and to successfully enter them; to become activated and proliferate; to exert various effector functions, including cytotoxic killing and the secretion of various factors such as cytokines; to survive (including the ability to survive for a long period); to differentiate into, transition to, or engage in reprogramming into certain phenotypic states; to provide a viable and robust recall response following clearance and re-exposure to target ligands or target antigens, thereby avoiding or reducing exhaustion, anergy, terminal differentiation, and / or differentiation into suppressive states.

[0226] In some cases, the available approaches for treating diseases or disorders such as multiple myeloma are complex and not always entirely satisfactory. In some cases, the choice of treatment regimen may depend on a number of factors, including drug availability, response to prior therapy, severity of relapse, eligibility for autologous stem cell transplantation (ASCT), and whether relapse occurred with or without therapy. In some cases, MM results in relapse and remission, and existing regimens may, in some cases, result in relapse and / or toxicity from treatment. In some cases, treatment can be particularly challenging for patients with malignant disease, e.g., persistent or relapsing disease after various therapies, high disease burden, e.g., high tumor burden, and / or particularly malignant types of disease, e.g., plasmacytoma, and the response to certain therapies in these patients may be poor or short-lived. In some cases, heavily pre-treated patients, e.g., those who have relapsed after several different prior therapies, may exhibit low response rates and / or high incidence of adverse events. In some aspects, the provided embodiments are based on the observation that treatment by the provided embodiments results in a high response rate, a low incidence of adverse events (e.g., toxicity), a prolonged response, and, in some cases, an improvement in response over time.

[0227] The embodiments provided are based on observations from clinical studies that, in some circumstances, administration of engineered cells expressing specific recombinant receptors, such as those described herein, results in high response rates and low rates of adverse events, such as cytokine release syndrome (CRS) or neurological events (NE; or neurotoxicity; NT). In some aspects, the cells, methods, and uses provided result in cell therapy exhibiting prolonged persistence of cells after administration, along with high response rates and low rates of toxicity (e.g., CRS or NE, e.g., grade 3 or higher CRS or grade 3 or higher neurotoxicity). In some aspects, such high response rates and low rates of toxicity (e.g., grade 3 or higher CRS or grade 3 or higher neurotoxicity) are achieved by using various different cell doses. For example, even with relatively low doses of cells, high rates of objective response and high levels of response (e.g., best partial response, VGPR, or better) are achieved. In some cases, relatively high doses of cells are administered, and such doses have been observed to result in high objective response rates along with low rates of toxicity (e.g., Grade 3 or higher CRS or Grade 3 or higher neurotoxicity). In some cases, the provided embodiments also enable improved expansion and / or persistence of the administered manipulated cells, resulting in prolonged responses and / or responses that improve over time. In some aspects, treatment of subjects with malignant or refractory diseases (e.g., severely pre-treated subjects, subjects with a high tumor burden and / or subjects with malignant disease species) by the provided embodiments has been observed to provide safe, effective, and sustained treatment.

[0228] In some situations, the optimal response to therapy may depend on the ability of engineered recombinant receptors, such as CARs, to be consistently and reliably expressed on the cell surface and / or to bind to the target antigen. For example, in some cases, heterogeneity of RNA transcribed from an introduced transgene (e.g., encoding a recombinant receptor) may affect the expression and / or activity of the recombinant receptor when expressed in cells used in cell therapy, such as human T cells. In some situations, the length and type of spacers in recombinant receptors such as CARs may affect the expression, activity, and / or function of the receptor.

[0229] Furthermore, in some situations, certain recombinant receptors may exhibit antigen-independent activity or signaling (also known as "tonic signaling"), which can lead to undesirable effects, such as increased differentiation and / or exhaustion of T cells expressing the recombinant receptor. In some cases, such activity may limit the activity, effect, or potency of T cells. In some cases, during cell manipulation for recombinant receptor expression and ex vivo expansion, cells may exhibit a phenotype indicating exhaustion, resulting from tonic signaling through the recombinant receptor.

[0230] In some situations, the characteristics of specific target antigens to which recombinant receptors specifically bind, recognize, or target can influence receptor activity. In some situations, B-cell maturation antigens (BCMAs), typically expressed on malignant plasma cells, are attractive therapeutic targets for cell therapy. In some cases, BCMAs can be cleaved by gamma secretases to produce soluble BCMAs (sBCMAs), or "shedded" forms of BCMAs, reducing the amount of BCMA expressed on the surface of target cells. In some cases, the activity of BCMA-binding molecules, such as anti-BCMA chimeric antigen receptors, can be blocked or inhibited by the presence of soluble BCMAs. Improved strategies are needed for optimal efficacy in cell therapy, particularly for recombinant receptors that specifically bind to, recognize, or target BCMAs, such as BCMAs expressed on the surface of target cells.

[0231] The embodiments provided are based on the observation that, in some circumstances, optimization of specific spacers and nucleic acid sequences can lead to consistent and robust expression of recombinant receptors. The BCMA-binding recombinant receptors provided offer advantages over available approaches for cell therapy, particularly BCMA-targeted cell therapy. In some embodiments, the BCMA-binding recombinant receptors provided contain a fully human antigen-binding domain with low affinity for binding to soluble BCMA. In some embodiments, the BCMA-binding recombinant receptors provided contain a modified spacer that results in enhanced binding to BCMA expressed on the surface of target cells. In some embodiments, the BCMA-binding recombinant receptors provided are observed to exhibit reduced antigen-independent tonic signaling, which in some cases may result in reduced cellular exhaustion from antigen-independent signaling and the absence of inhibition by soluble BCMA. In some embodiments, the BCMA-binding recombinant receptors provided exhibit activity or efficacy against target cells expressing low densities or low levels of BCMA.

[0232] In various aspects, the BCMA-binding recombinant receptors, polynucleotides encoding such receptors, engineered cells, and cell compositions provided herein can overcome or counteract certain limitations that may reduce the optimal response to cell therapy, such as cell therapy using engineered cells expressing BCMA-binding recombinant receptors, exhibiting certain desired properties. In some aspects, compositions containing engineered cells expressing exemplary BCMA-binding recombinant receptors provided herein have been observed to exhibit consistency in the cellular health of the engineered cells and have been associated with improved clinical response. In some aspects, compositions containing engineered cells expressing exemplary BCMA-binding recombinant receptors provided herein have been associated in some aspects with increased persistence and durability of the engineered cells, particularly with central memory T cells (T CM ) Enrichment of immune cell subtypes associated with the phenotype, e.g., CD4+ or CD8+ T cell subtypes, has been observed. In some situations, embodiments provided, comprising recombinant receptors, polynucleotides encoding such receptors, engineered cells, and cell compositions, can offer various advantages over available BCMA-targeted therapies to enhance the activity of recombinant receptors and the response to BCMA-targeted cell therapy. In addition, methods and uses provided of engineered cells or compositions comprising engineered cells have been observed to offer advantages in the treatment of subjects, resulting in high response rates, sustained responses, and low rates of adverse events at various different dose levels tested. Furthermore, methods and uses provided of engineered cells or compositions comprising engineered cells have been observed to offer advantages in the treatment of subjects with malignant and / or refractory diseases, or subjects that have relapsed and / or are refractory to a number of different prior treatments for the disease.

[0233] A. Chimeric antigen receptor BCMA conjugates that bind to or recognize the BCMA molecule, such as cell surface proteins, such as recombinant receptors or chimeric antigen receptors, and BCMA-binding cell surface proteins, such as polynucleotides encoding recombinant receptors (e.g., chimeric antigen receptors; CARs), and cells expressing such receptors are provided in some aspects. BCMA-binding cell surface proteins generally contain antibodies (e.g., antigen-binding antibody fragments) and / or other binding peptides that specifically recognize, or specifically bind to, BCMA, such as BCMA proteins, such as human BCMA proteins. In some aspects, these conjugates bind to the extracellular portion of BCMA. Cells containing such polynucleotides or expressing such receptors, such as engineered cells, and compositions containing such engineered cells are also provided. In some aspects, methods using such cells and compositions, as well as their use in therapeutic methods, etc., are also provided.

[0234] In some embodiments, polynucleotides are optimized, or contain certain features designed for optimization, to reduce RNA heterogeneity and / or modify, for example, increased or more consistent, the expression of the encoded receptor, e.g., surface expression, or to make it more consistent between cell product lots, for example, with respect to codon usage frequency. In some embodiments, polynucleotides encoding BCMA-binding cell surface proteins are modified to reduce RNA heterogeneity, for example, by removing latent or hidden splice sites, compared to a reference polynucleotide. In some embodiments, polynucleotides encoding BCMA-binding cell surface proteins are codon-optimized for expression in mammalian cells, e.g., human cells, e.g., human T cells. In some aspects, modified polynucleotides, when expressed in cells, result in improved, e.g., increased or more uniform or more consistent levels of expression, e.g., surface expression. Such polynucleotides can be utilized in constructs for the generation of engineered cells expressing the encoded BCMA-binding cell surface protein. Therefore, cells expressing recombinant receptors encoded by polynucleotides provided herein, as well as adoptive cell therapies, are also provided, for example, for diseases and disorders associated with BCMA expression, such as multiple myeloma.

[0235] Among the polynucleotides provided are those that encode recombinant receptors, such as antigen receptors, that specifically recognize or bind to BCMA, such as human BCMA. In some aspects, the encoded receptors, such as those containing BCMA-binding polypeptides, as well as their compositions, products, and uses, are also provided.

[0236] BCMA-binding polypeptides include antibodies, such as single-chain antibodies (e.g., antigen-binding antibody fragments), or portions thereof. In some examples, the recombinant receptor is a chimeric antigen receptor, such as an anti-BCMA antibody or a chimeric antigen receptor containing an antigen-binding fragment thereof. In any embodiment, the antibody or antigen-binding fragment in the provided CAR, which specifically recognizes an antigen, such as BCMA, specifically binds to the antigen. The provided polynucleotides can be incorporated into constructs, such as deoxyribonucleic acid (DNA) or RNA constructs, which can be introduced into cells for the expression of a recombinant BCMA-binding receptor encoding such a receptor.

[0237] In some cases, the polynucleotide encoding the BCMA-binding receptor contains a signal sequence encoding a signal peptide, which in some cases is encoded upstream of the nucleic acid sequence encoding the BCMA-binding receptor or is ligated at the 5' end of the nucleic acid sequence encoding the antigen-binding domain. In some cases, the polynucleotide containing the nucleic acid sequence encoding the BCMA-binding receptor, e.g., a chimeric antigen receptor (CAR), contains a signal sequence encoding 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 heterogeneous or non-native signal peptide. In some cases, the polynucleotide encoding the BCMA-binding receptor may contain an additional molecule, e.g., a nucleic acid sequence encoding a surrogate marker or other marker, or an additional component, e.g., a promoter, regulatory element, and / or a multicistronic element. In some embodiments, the nucleic acid sequence encoding the BCMA-binding receptor may be functionally ligated to any of the additional components.

[0238] For example, the BCMA-binding receptors provided, expressed in cells used in the methods and uses provided herein, generally contain extracellular binding molecules and intracellular signaling domains. Among the binding molecules provided are antibody-containing polypeptides, such as single-chain cell surface proteins containing such antibodies, for example, recombinant receptors, for example, chimeric antigen receptors.

[0239] The binding molecules provided (e.g., BCMA-binding molecules) include single-chain cell surface proteins, such as recombinant receptors (e.g., antigen receptors), containing one of the provided antibodies or fragments thereof (e.g., BCMA-binding fragments). Recombinant receptors include antigen receptors that specifically bind to or recognize BCMA, such as the provided anti-BCMA antibodies, such as antigen receptors containing antigen-binding fragments. Antigen receptors include functional non-TCR antigen receptors, such as chimeric antigen receptors (CARs). Cells expressing recombinant receptors and their use in adoptive cell therapy, such as in diseases and disorders associated with BCMA expression, are also provided.

[0240] Exemplary antigen receptors, including CARs, and methods for manipulating and introducing such antigen receptors into cells, are, for example, International Patent Application Publications WO200014257, WO2013126726, WO2012 / 129514, WO2014031687, WO2013166321, WO2013071154, WO2013123061, U.S. Patent Application Publications US2002131960, US2013287748, US20130149337, U.S. Patent Publications 6,4 Patent Nos. 51,995, 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 those described in European Patent Application EP2537416, and / or Sadelain Examples include those described in et al., Cancer Discov. 2013 April;3(4):388-398; Davida et al. (2013) PLoS ONE 8(4):e61338; Turtle et al., Curr. Opin. Immunol., 2012 October;24(5):633-39; and Wu et al., Cancer, 2012 March 18(2):160-75. In some aspects, examples of antigen receptors include the CAR described in U.S. Patent No. 7,446,190 and those described in International Patent Application Publication WO2014055668. Examples of CARs include those disclosed in any of the aforementioned publications, e.g., WO2014031687, US 8,339,645, US 7,446,179, US 2013 / 0149337, US 7,446,190, and US 8,389,282, in which the antigen-binding moiety, e.g., scFv, is replaced by an antibody or its antigen-binding fragment provided herein.

[0241] In some embodiments, the provided CAR has an amino acid sequence that exhibits at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with an amino acid sequence selected from SEQ ID NO: 15-20, or an amino acid sequence described in any of SEQ ID NO: 15-20, or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity. In some embodiments, the provided CAR has an amino acid sequence that exhibits at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with respect to the amino acid sequence described in SEQ ID NO:19, or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity.

[0242] In some embodiments, the provided CAR is encoded by a polynucleotide, for example, a nucleic acid sequence described in any of SEQ ID NO: 9-14, or a polynucleotide having a sequence that exhibits at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with a nucleic acid sequence described in any of SEQ ID NO: 9-14, or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity. In some embodiments, the provided CAR is encoded by a polynucleotide, for example, a nucleic acid sequence described in either SEQ ID NO: 13 or 14, or a polynucleotide having a sequence that exhibits at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with respect to the nucleic acid sequence described in either SEQ ID NO: 13 or 14, or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity. In some embodiments, the provided CAR is encoded by a polynucleotide, for example, a nucleic acid sequence described in SEQ ID NO:13, or a polynucleotide having a sequence that exhibits at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, 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, for example, a polynucleotide having a nucleic acid sequence described in SEQ ID NO:13.

[0243] In some embodiments, the nucleic acid encoding the antigen-binding domain includes (a) a sequence of nucleotides listed in any of SEQ ID NO: 30, 31, 50, 51, 59, 60, 82, 84, 113, 115, (b) a sequence of nucleotides having at least 90% sequence identity to any of SEQ ID NO: 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 includes (a) a sequence of nucleotides encoding an amino acid sequence described in any of SEQ ID NO: 29, 49, 58, 83, 114, 127, 128, 129, 130, (b) a sequence of nucleotides having at least 90% sequence identity with respect to a sequence of nucleotides encoding an amino acid sequence described in any of SEQ ID NO: 29, 49, 58, 83, 114, 126, 127, 128, 129, 130, or (c) a degenerate sequence of (a) or (b).

[0244] 1. Antigen-binding domain Among chimeric receptors are chimeric antigen receptors (CARs). Chimeric receptors, such as CARs, generally contain, are, or are contained within one of the provided anti-BCMA antibodies, or contain an extracellular antigen-binding domain that constitutes it. Therefore, chimeric receptors, such as CARs, typically contain in their extracellular component one or more BCMA-binding molecules, such as one or more antigen-binding fragments, domains, or portions, or one or more antibody variable regions, and / or antibody molecules, such as those described herein.

[0245] The term “antibody” is used herein in its broadest sense and includes polyclonal and monoclonal antibodies, including intact antibodies and functional (antigen-binding) antibody fragments, such fragments including antigen-binding fragment (Fab) fragments, F(ab')2 fragments, Fab' fragments, Fv fragments, recombinant IgG (rIgG) fragments, heavy chain variable (VH) regions capable of specifically binding to antigens, single-chain antibody fragments including single-chain variable fragments (scFv), and single-domain antibody (e.g., sdAb, sdFv, nanobody) fragments. The term also includes genetically engineered and / or otherwise modified forms of immunoglobulins, such as intrabodies, peptidebodies, chimeric antibodies, fully human antibodies, humanized antibodies, and heteroconjugate antibodies, multispecific antibodies, such as bispecific or tripspecific antibodies, diabodies, triabodies, and tetrabodies, tandem di-scFv, and tandem tri-scFv. Unless otherwise specified, the term “antibody” should be understood herein to encompass its functional antibody fragment, also referred to herein as “antigen-binding fragment.” This term also encompasses intact or full-length antibodies, including antibodies of any class or subclass, including IgG and its subclasses, IgM, IgE, IgA, and IgD.

[0246] The terms "complementarity-determining region" and "CDR" are synonymous with "hypervariable region" or "HVR," and are known in the art to refer to discontinuous sequences of amino acids within the antibody variable region that confer antigen specificity and / or binding affinity. Generally, each heavy chain variable region has three CDRs (CDR-H1, CDR-H2, CDR-H3), and each light chain region has three CDRs (CDR-L1, CDR-L2, CDR-L3). The terms "framework region" and "FR" are known in the art to refer to the non-CDR portions of the heavy and light chain variable regions. Generally, each full-length heavy chain variable region has four FRs (FR-H1, FR-H2, FR-H3, and FR-H4), and each full-length light chain variable region has four FRs (FR-L1, FR-L2, FR-L3, and FR-L4).

[0247] The precise amino acid sequence boundaries of a given CDR or FR can be easily identified using one of several well-known methods. These methods include: Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (“Kabat” numbering method); Al-Lazikani et al., (1997) JMB 273,927-948 (“Chothia” numbering method); 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 method); 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 This includes the "IMGT" numbering scheme; the "Aho" numbering scheme; and the scheme described by Honegger A and Pluckthun 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; and the "AbM" numbering scheme; and Martin et al., “Modeling antibody hypervariable loops: a combined algorithm,” PNAS, 1989, 86(23):9268-9272.

[0248] The boundaries of a given CDR or FR can vary depending on the specific method used. For example, the Kabat method is based on structural alignment, while the Chothia method is based on structural information. Both Kabat and Chothia numbering are based on the most common antibody region sequence lengths, with insertions corresponding by insertion letters, e.g., "30a," and deletions appearing in some antibodies. These two methods assign different numbers because they place certain insertions and deletions ("indels") in different positions. The Contact method is based on the analysis of the complex crystal structure and is similar in many ways to the Chothia numbering method. The AbM method is a compromise between the Kabat and Chothia definitions, based on what is used by Oxford Molecular's AbM antibody modeling software.

[0249] Table 6 below lists exemplary positional boundaries of CDR-L1, CDR-L2, CDR-L3 and CDR-H1, CDR-H2, CDR-H3 as identified by the Kabat, Chothia, AbM, and Contact schemes, respectively. For CDR-H1, residue numbering is described using both the Kabat and Chothia numbering schemes. FRs are located between CDRs; for example, FR-L1 is located anterior to CDR-L1, FR-L2 is located between CDR-L1 and CDR-L2, FR-L3 is located between CDR-L2 and CDR-L3, and so on. Note that because the Kabat numbering scheme shown places insertions at H35A and H35B, the end of the Chothia CDR-H1 loop will vary between H32 and H34 depending on the loop length, when numbered using the conventions of the Kabat numbering scheme shown.

[0250] (Table 6) Boundaries of CDRs based on various numbering schemes TIFF2026113544000009.tif471611 - 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

[0251] Therefore, unless otherwise specified, a given antibody or region, for example, its variable region's "CDR" or "complementarity-determining region," or individual designated CDRs (e.g., CDR-H1, CDR-H2, CDR-H3), should be understood to encompass a certain (or specific) complementarity-determining region as defined by one of the aforementioned methods or other known methods. For example, a particular CDR (e.g., CDR-H3) may be a given V H Area or V L Where it is stated that a region contains the amino acid sequence of a corresponding CDR, such a CDR is understood to have the sequence of the corresponding CDR (e.g., CDR-H3) within its variable region, as defined by one of the aforementioned methods or other known methods. In some embodiments, specific CDR sequences are specified. Exemplary CDR sequences of the antibodies provided are described using various numbering schemes, but it is understood that the antibodies provided may contain CDRs as described according to one of the other aforementioned numbering schemes or other numbering schemes known to those skilled in the art.

[0252] Similarly, unless otherwise specified, a given antibody or its region, for example, its variable region FR or individual designated FRs (e.g., FR-H1, FR-H2, FR-H3, FR-H4), should be understood to encompass a certain (or specific) framework region defined by one of the known methods. In some cases, a method for identifying individual CDRs, FRs, or multiple FRs or CDRs is specified. For example, a CDR may be defined by the Kabat, Chothia, AbM, IMGT, or Contact method, or by another known method. In other cases, a specific amino acid sequence of a CDR or FR is given.

[0253] The term "variable region" or "variable domain" refers to a domain in the antibody heavy or light chain that is involved in the binding of an antibody to an antigen. The variable regions of the native antibody heavy and light chains (V, respectively) H and V L ) generally have a similar structure in which each domain contains four conserved framework regions (FRs) and three CDRs (see, for example, Kindt et al. Kuby Immunology, 6th ed., WH Freeman and Co., page 91 (2007)). Single V H or V L The domain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind to a specific antigen may each have complementary V L or V H To screen the domain library, V derived from antibodies that bind to the antigen is used. H or V L They may be isolated using domains. See, for example, Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0254] Some of the antibodies contained in the provided CAR include antibody fragments. An "antibody fragment" or "antigen-binding fragment" refers to a molecule other than the intact antibody, containing a portion of the intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibodies, but not limited to these, include Fv, Fab, Fab', Fab'-SH, F(ab')2 diabodies; linear antibodies; and heavy-chain variable (V) antibodies. H ) region, single-chain antibody molecule, e.g., scFv, and V H Examples include single-domain antibodies containing only a region, as well as multispecific antibodies formed from antibody fragments. In some embodiments, the antigen-binding domain in the provided CAR is a variable heavy chain (V H ) and variable light chain (V L The antibody fragment contains or includes the ) region. In certain embodiments, the antibody is heavy chain variable (V H ) region and / or light chain variable (V L This is a single-chain antibody fragment containing the ) region, for example, scFv.

[0255] A single-domain antibody (sdAb) is an antibody fragment that contains all or part of the heavy chain variable region or all or part of the light chain variable region of an antibody. In certain embodiments, a single-domain antibody is a human single-domain antibody.

[0256] Antibody fragments can be produced by a variety of techniques, including, but not limited to, proteolytic digestion of intact antibodies and production by recombinant host cells. In some embodiments, antibodies are fragments produced by recombination, such as fragments containing configurations not occurring in nature, such as fragments in which two or more antibody regions or chains are linked by a synthetic linker, such as a peptide linker, and / or fragments that may not be produced by enzymatic digestion of naturally occurring intact antibodies. In some aspects, antibody fragments are scFv.

[0257] A “humanized” antibody is one in which all or substantially all CDR amino acid residues are derived from non-human CDRs and all or substantially all FR amino acid residues are derived from human FRs. A humanized antibody may optionally contain at least a portion of the antibody constant region der...

Claims

1. A method for treating multiple myeloma (MM) in a subject having or suspected of having MM, comprising the step of administering a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) targeting B cell maturation antigen (BCMA), The composition is a CD8 that expresses the CAR. ＋ T cells and CD4 expressing the CAR ＋ Includes T cells, The composition, including the values ​​at both ends, is 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~80 × 10⁶ 6 1 or approximately 80 x 10 6 Contains CAR-expressing T cells, At least 80% or at least about 80% of the cells in the composition are CD3 ＋ Cells are healthy, and CAR in the composition ＋ At least 80% or at least about 80% of T cells are naive-like or central memory phenotype cells. The aforementioned method.

2. A method for treating multiple myeloma (MM) in a subject having or suspected of having MM, comprising the step of administering a composition comprising engineered T cells expressing a chimeric antigen receptor (CAR) targeting B cell maturation antigen (BCMA), The composition comprises CD8 ＋ T cells expressing the CAR and CD4 ＋ T cells expressing the CAR, and The composition, including the values ​​at both ends, is 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~ 100 × 10⁶ 6 1 or approximately 100 x 10 6 Contains CAR-expressing T cells, At least 80% or at least about 80% of the cells in the composition are CD3 ＋ It is a cell, and CD4 in the composition ＋ CAR ＋ At least 50% or at least about 50% of T cells are CD27 ＋ CCR7 ＋ and / or CD8 in the composition ＋ CAR ＋ At least 50% or at least about 50% of T cells are CD27 ＋ CCR7 ＋ That is, The aforementioned method.

3. The composition is a CD4 that expresses CAR. ＋ T cells and CD8 expressing CAR ＋ The method according to claim 1 or claim 2, comprising T cells in a ratio of approximately 1:2.5 to approximately 5:

1.

4. The composition is a CD4 that expresses CAR. ＋ T cells and CD8 expressing CAR ＋ The method according to any one of claims 1 to 3, comprising T cells in a ratio of approximately 1:2 to approximately 4:1, approximately 1:1.5 to approximately 2:1, or 1:1 or approximately 1:

1.

5. The composition is a CD4 that expresses CAR. ＋ T cells and CD8 expressing CAR ＋ The method according to any one of claims 1 to 3, comprising T cells in a ratio of approximately 5:1 to approximately 2:1, approximately 4:1 to approximately 2:1, approximately 3:1 to approximately 2:1, 5:1 or approximately 5:1, 4:1 or approximately 4:1, 3:1 or approximately 3:1, or 2:1 or approximately 2:

1.

6. The composition, including the values ​​at both ends, is 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~80 × 10⁶ 6 1 or approximately 80 x 10 6 The method according to any one of claims 2 to 5, comprising CAR-expressing T cells.

7. The composition, including the values ​​at both ends, is 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~40 × 10⁶ 6 1 or approximately 40 x 10 6 The method according to any one of claims 1 to 6, comprising CAR-expressing T cells.

8. The composition, including the values ​​at both ends, is 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~20 × 10⁶ 6 1 or approximately 20 x 10 6 The method according to any one of claims 1 to 7, comprising CAR-expressing T cells.

9. The composition, including the values ​​at both ends, is 5 × 10 6 pc or approximately 5 x 10 6 Individual CAR-expressing T cells ~ 10 × 10 6 1 or approximately 10 x 10 6 The method according to any one of claims 1 to 8, comprising CAR-expressing T cells.

10. The composition, including the values ​​at both ends, is 10 × 10 6 1 or approximately 10 x 10 6 Individual CAR-expressing T cells ~20 × 10⁶ 6 1 or approximately 20 x 10 6 The method according to any one of claims 1 to 8, comprising CAR-expressing T cells.

11. The composition is 20 × 10 6 1 or approximately 20 x 10 6 The method according to any one of claims 1 to 8 and 10, comprising CAR-expressing T cells.

12. The composition is 30 × 10 6 1 or approximately 30 x 10 6 The method according to any one of claims 1 to 7, comprising CAR-expressing T cells.

13. The composition is 40 × 10 6 1 or approximately 40 x 10 6 The method according to any one of claims 1 to 7, comprising CAR-expressing T cells.

14. At least 90% or at least about 90% of the cells in the composition are CD3 ＋ The method according to any one of claims 1 to 13, wherein the cell is a cell.

15. At least 91% or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, or at least 96% or at least about 96% of the cells in the composition are CD3 ＋ The method according to any one of claims 1 to 14, wherein the cell is a cell.

16. CAR in the composition ＋ The method according to any one of claims 1 to 15, wherein 2% or about 2% to 30% or about 30% of the T cells express an apoptosis marker, optionally annexin V or active caspase 3.

17. CAR in the composition ＋ The method according to any one of claims 1 to 16, wherein 5% or about 5% to 10% or about 10% of the T cells express an apoptosis marker, optionally annexin V or active caspase 3.

18. CAR in the composition ＋ The method according to any one of claims 1 to 16, wherein 10% or about 10% to 15% or about 15% of the T cells express an apoptosis marker, optionally annexin V or active caspase 3.

19. CAR in the composition ＋ The method according to any one of claims 1 to 16, wherein 15% or about 15% to 20% or about 20% of the T cells express an apoptosis marker, optionally annexin V or active caspase 3.

20. CAR in the composition ＋ The method according to any one of claims 1 to 16, wherein 20% or about 20% to 30% or about 30% of the T cells express an apoptosis marker, optionally annexin V or active caspase 3.

21. CAR in the composition ＋ The method according to any one of claims 1 to 16, wherein 5% or about 5%, 10% or about 10%, 15% or about 15%, 20% or about 20%, 25% or about 25%, or 30% or about 30% of the T cells express an apoptosis marker, optionally annexin V or active caspase 3.

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

23. CAR in the composition ＋ The method according to any one of claims 1 to 22, wherein 80% or about 80% to 85% or about 85% of the T cells are naive-like or central memory phenotype cells.

24. CAR in the composition ＋ The method according to any one of claims 1 to 22, wherein 85% or about 85% to 90% or about 90% of the T cells are naive-like or central memory phenotype cells.

25. CAR in the composition ＋ The method according to any one of claims 1 to 22, wherein 90% or about 90% to 95% or about 95% of the T cells are naive-like or central memory phenotype cells.

26. CAR in the composition ＋ The method according to any one of claims 1 to 22, wherein 95% or about 95% to 99% or about 99% of the T cells are naive-like or central memory phenotype cells.

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

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

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

30. CAR in a composition of cells with a naive-like or central memory phenotype ＋ At least 80% or at least about 80% of T cells are CCR7 ＋ CD45RA ＋ CD27 ＋ CCR7 ＋ Or CD62L - CCR7 ＋ The method according to any one of claims 1 and 3 to 29, having a phenotype selected from among.

31. CAR in the composition ＋ Of the T cells, 80% or about 80%, 85% or about 85%, 85% or about 85%, 90% or about 90%, 90% or about 90%, 95% or about 95%, 95% or about 95%, and 99% or about 99% are CCR7 ＋ CD45RA ＋ CD27 ＋ CCR7 ＋ Or CD62L - CCR7 ＋ The method according to any one of claims 1 to 30, wherein the cells are selected from naive-like or central memory phenotypes.

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

33. CAR in the composition ＋ 80% or approximately 80%, 85% or approximately 85%, 90% or approximately 90%, 95% or approximately 95%, or 99% or approximately 99% of T cells are CD27 ＋ CCR7 ＋ The method according to any one of claims 1 to 32, wherein the cells are naive-like or have a central memory phenotype.

34. CD4 in the composition ＋ CAR ＋ At least 50% or at least about 50% of the T cells are CCR7 ＋ CD45RA ＋ Or CCR7 ＋ CD45RA - The method according to any one of claims 1 to 33, which is a cell of a naive or central memory phenotype that is

35. CD4 in the composition ＋ CAR ＋ At least 60% or at least about 60% of T cells are CCR7 ＋ CD45RA ＋ or CCR7 ＋ CD45RA - The method according to any one of claims 1 to 34, wherein the cells are naive-like or have a central memory phenotype.

36. CD4 in the composition ＋ CAR ＋ At least 70% or at least about 70% of T cells are CCR7 ＋ CD45RA ＋ or CCR7 ＋ CD45RA - The method according to any one of claims 1 to 35, wherein the cells are naive-like or have a central memory phenotype.

37. CD4 in the composition ＋ CAR ＋ At least 80% or at least about 80% of T cells are CCR7 ＋ CD45RA ＋ or CCR7 ＋ CD45RA - The method according to any one of claims 1 to 36, wherein the cells are naive-like or have a central memory phenotype.

38. CD4 in the composition ＋ CAR ＋ At least 85% or at least about 85% of T cells are CCR7 ＋ CD45RA ＋ or CCR7 ＋ CD45RA - The method according to any one of claims 1 to 37, wherein the cells are naive-like or have a central memory phenotype.

39. CD4 in the composition ＋ CAR ＋ At least 50% or at least about 50% of T cells are CD27 ＋ CCR7 ＋ The method according to any one of claims 1 to 38, wherein the cells are naive-like or have a central memory phenotype.

40. CD4 in the composition ＋ CAR ＋ At least 60% or at least about 60% of T cells are CD27 ＋ CCR7 ＋ The method according to any one of claims 1 to 39, wherein the cells are naive-like or have a central memory phenotype.

41. CD4 in the composition ＋ CAR ＋ At least 70% or at least about 70% of T cells are CD27 ＋ CCR7 ＋ The method according to any one of claims 1 to 40, wherein the cells are naive-like or have a central memory phenotype.

42. CD4 in the composition ＋ CAR ＋ At least 80% or at least about 80% of T cells are CD27 ＋ CCR7 ＋ The method according to any one of claims 1 to 41, wherein the cells are naive-like or have a central memory phenotype.

43. CD4 in the composition ＋ CAR ＋ At least 85% or at least about 85% of T cells are CD27 ＋ CCR7 ＋ The method according to any one of claims 1 to 42, wherein the cells are naive-like or have a central memory phenotype.

44. CD8 in the composition ＋ CAR ＋ At least 50% or at least about 50% of T cells are CCR7 ＋ CD45RA ＋ or CCR7 ＋ CD45RA - The method according to any one of claims 1 to 43, wherein the cells are naive-like or have a central memory phenotype.

45. CD8 in the composition ＋ CAR ＋ At least 60% or at least about 60% of T cells are CCR7 ＋ CD45RA ＋ or CCR7 ＋ CD45RA - The method according to any one of claims 1 to 44, wherein the cells are naive-like or have a central memory phenotype.

46. CD8 in the composition ＋ CAR ＋ At least 70% or at least about 70% of T cells are CCR7 ＋ CD45RA ＋ or CCR7 ＋ CD45RA - The method according to any one of claims 1 to 45, wherein the cells are naive-like or have a central memory phenotype.

47. CD8 in the composition ＋ CAR ＋ At least 80% or at least about 80% of T cells are CCR7 ＋ CD45RA ＋ or CCR7 ＋ CD45RA - The method according to any one of claims 1 to 46, wherein the cells are naive-like or have a central memory phenotype.

48. CD8 in the composition ＋ CAR ＋ At least 85% or at least about 85% of T cells are CCR7 ＋ CD45RA ＋ or CCR7 ＋ CD45RA - The method according to any one of claims 1 to 47, wherein the cells are naive-like or have a central memory phenotype.

49. CD8 in the composition ＋ CAR ＋ At least 50% or at least about 50% of T cells are CD27 ＋ CCR7 ＋ The method according to any one of claims 1 to 48, wherein the cells are naive-like or have a central memory phenotype.

50. CD8 in the composition ＋ CAR ＋ At least 60% or at least about 60% of T cells are CD27 ＋ CCR7 ＋ The method according to any one of claims 1 to 49, wherein the cells are naive-like or have a central memory phenotype.

51. CD8 in the composition ＋ CAR ＋ At least 70% or at least about 70% of T cells are CD27 ＋ CCR7 ＋ The method according to any one of claims 1 to 50, wherein the cells are naive-like or have a central memory phenotype.

52. CD8 in the composition ＋ CAR ＋ At least 80% or at least about 80% of T cells are CD27 ＋ CCR7 ＋ The method according to any one of claims 1 to 51, wherein the cells are naive-like or have a central memory phenotype.

53. CD8 in the composition ＋ CAR ＋ At least 85% or at least about 85% of T cells are CD27 ＋ CCR7 ＋ The method according to any one of claims 1 to 52, wherein the cells are naive-like or have a central memory phenotype.

54. CAR in the composition ＋ The method according to any one of claims 1 to 53, wherein the fraction of the number of incorporated vectors (iVCNs) to the total number of vector copies (VCNs) in T cells is, on average, less than 0.9 or about 0.

9.

55. CAR in the composition ＋ The method according to any one of claims 1 to 54, wherein the fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, 0.9 or about 0.9 to 0.8 or about 0.

8.

56. CAR in the composition ＋ The method according to any one of claims 1 to 54, wherein the fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, less than 0.8 or about 0.

8.

57. CAR in the composition ＋ The method according to any one of claims 1 to 54 and 56, wherein the fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, 0.8 or about 0.8 to 0.7 or about 0.

7.

58. CAR in the composition ＋ The method according to any one of claims 1 to 54 and 56, wherein the fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, 0.7 or about 0.7 to 0.6 or about 0.

6.

59. CAR in the composition ＋ The method according to any one of claims 1 to 54 and 56, wherein the fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, 0.6 or about 0.6 to 0.5 or about 0.

5.

60. CAR in the composition ＋ The method according to any one of claims 1 to 54 and 56, wherein the fraction of the copy number of the incorporated vector (iVCN) relative to the total VCN in T cells is, on average, 0.5 or about 0.5 to 0.4 or about 0.

4.

61. CAR in the composition ＋ The method according to any one of claims 1 to 60, wherein the copy number (iVCN) of the vector into which the T cells are incorporated is, on average, 0.4 copies to 2.0 copies per diploid genome, including the values ​​at both ends, or approximately 0.4 copies to approximately 2.0 copies per diploid genome.

62. CAR in the composition ＋ The method according to any one of claims 1 to 61, wherein the copy number (iVCN) of the vector into which the T cells are incorporated is, on average, 0.8 copies to 2.0 copies per diploid genome, including the values ​​at both ends, or approximately 0.8 copies to approximately 2.0 copies per diploid genome.

63. CAR in the composition ＋ The method according to any one of claims 1 to 62, wherein the copy number (iVCN) of the vector into which the T cells are incorporated is, on average, 0.8 copies to 1.0 copy per diploid genome, including the values ​​at both ends, or approximately 0.8 copies to approximately 1.0 copy per diploid genome.

64. CAR in the composition ＋ The method according to any one of claims 1 to 62, wherein the copy number (iVCN) of the vector into which the T cells are incorporated is, on average, 1.0 copy to 1.5 copies per diploid genome, or about 1.0 copy to about 1.5 copies per diploid genome, including the values ​​at both ends.

65. CAR in the composition ＋ The method according to any one of claims 1 to 62, wherein the copy number (iVCN) of the vector into which the T cells are incorporated is, on average, 1.5 copies to 2.0 copies per diploid genome, including the values ​​at both ends, or approximately 1.5 copies to approximately 2.0 copies per diploid genome.

66. The method according to any one of claims 1 to 65, wherein the subject has received at least three prior anti-myeloma treatment regimens at the time of or before administration of the composition containing the manipulated T cells.

67. The method according to any one of claims 1 to 66, wherein, at the time of or prior to administration of a composition containing manipulated T cells, the subject has received all three of the following anti-myeloma treatment regimens: autologous stem cell transplantation (ASCT), a regimen comprising an immunomodulator and a proteasome inhibitor, and an anti-CD38 agent.

68. The method according to claim 66 or 67, wherein the subject is refractory to the last anti-myeloma treatment regimen at the time of or before administration of the composition containing manipulated T cells.

69. The method according to claim 68, wherein refractory myeloma is defined as a confirmed progressive disease during treatment with the last antimyeloma treatment regimen, or within 12 months from the last dose after the completion of such treatment.

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

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

72. The method according to any one of claims 1 to 71, further comprising the step of obtaining a leukocyte apheresis sample from a subject in order to produce a composition containing manipulated T cells.

73. CAR, (a) Variable heavy chain (V) containing heavy chain complementarity determination region 1 (CDR-H1), heavy chain complementarity determination region 2 (CDR-H2), and heavy chain complementarity determination region 3 (CDR-H3) within the sequence shown in SEQ ID NO:116 H ), as well as the variable light chain (V) containing light chain complementarity determination region 1 (CDR-L1), light chain complementarity determination region 2 (CDR-L2), and light chain complementarity determination region 3 (CDR-L3) within the sequence shown in SEQ ID NO:

119. L ); V containing the CDR-H1, CDR-H2, and CDR-H3 sequences shown in SEQ ID NO: 97, 101, and 103, respectively. H , and V containing the CDR-L1, CDR-L2, and CDR-L3 sequences shown in SEQ ID NO: 105, 107, and 108, respectively. L ; V containing the CDR-H1, CDR-H2, and CDR-H3 sequences shown in SEQ ID NO: 96, 100, and 103, respectively. H , and V containing the CDR-L1, CDR-L2, and CDR-L3 sequences shown in SEQ ID NO: 105, 107, and 108, respectively. L ; V containing the CDR-H1, CDR-H2, and CDR-H3 sequences shown in SEQ ID NO: 95, 99, and 103, respectively. H , and V containing the CDR-L1, CDR-L2, and CDR-L3 sequences shown in SEQ ID NO: 105, 107, and 108, respectively. L ; V containing the CDR-H1, CDR-H2, and CDR-H3 sequences shown in SEQ ID NO: 94, 98, and 102, respectively. H , and V containing the CDR-L1, CDR-L2, and CDR-L3 sequences shown in SEQ ID NO: 104, 106, and 108, respectively. L ;or V containing the amino acid sequence of SEQ ID NO:116 H , and V containing the amino acid sequence of SEQ ID NO:119 L Extracellular antigen-binding domain including, (b) IgG4 / 2 chimeric hinge or modified IgG4 hinge, IgG2 / 4 chimeric C H 2 regions, and IgG4 C H A spacer containing 3 regions and optionally approximately 228 amino acid lengths, and optionally the spacer shown in SEQ ID NO:

174. (c) Transmembrane domain, optionally human CD28-derived transmembrane domain, and (d) An intracellular signaling region comprising the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain and a costimulatory signaling region that includes the intracellular signaling domain or signaling portion of a T cell costimulatory molecule. A method according to any one of claims 1 to 72, including the method described in any one of claims 1 to 72.

74. V H is the amino acid sequence of SEQ ID NO:116 or contains the amino acid sequence of SEQ ID NO:116, V L The method according to claim 73, wherein is the amino acid sequence of SEQ ID NO:119 or comprises the amino acid sequence of SEQ ID NO:

119.

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

76. V H and V L The method according to any one of claims 73 to 75, wherein the two are connected by a flexible linker.

77. scFv is an amino acid sequence The method according to claim 75 or claim 76, comprising a linker containing the linker.

78. The method according to any one of claims 73 to 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 with respect to the amino acid sequence of SEQ ID NO:

114.

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

114.

80. The method according to any one of claims 73 to 79, wherein the cytoplasmic signaling domain is the sequence shown in SEQ ID NO:143 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:143, or comprises the sequence shown in SEQ ID NO:143 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:

143.

81. The method according to any one of claims 73 to 80, wherein the co-stimulatory signaling region includes an intracellular signaling domain or signaling portion of CD28, 4-1BB, or ICOS.

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

83. The method according to any one of claims 73 to 82, wherein the co-stimulatory signaling region is the sequence shown in SEQ ID NO:4 or an amino acid sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO:4, or includes the sequence shown in SEQ ID NO:4 or an amino acid sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO:

4.

84. The method according to any one of claims 73 to 83, wherein the co-stimulatory signaling region is located between the transmembrane domain and the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain.

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

86. The method according to any one of claims 73 to 85, wherein the transmembrane domain is the sequence shown in SEQ ID NO:138 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:138, or comprises the sequence shown in SEQ ID NO:138 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:

138.

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

88. CAR, (a) An extracellular antigen-binding domain comprising the sequence shown in SEQ ID NO:114 or an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO:114, (b) A spacer comprising the sequence shown in SEQ ID NO:174 or an amino acid sequence having at least 90% sequence identity with SEQ ID NO:174, (c) A transmembrane domain comprising the sequence shown in SEQ ID NO:138 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:138, and (d) An intracellular signaling region comprising a cytoplasmic signaling region including the sequence shown in SEQ ID NO:143 or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:143, and a co-stimulatory signaling region including the sequence shown in SEQ ID NO:4 or an amino acid sequence having at least 90% sequence identity to the sequence shown in SEQ ID NO:

4. A method according to any one of claims 1 to 87, including the method described in any one of claims 1 to 87.

89. CAR, (a) Extracellular antigen-binding domain containing the sequence shown in SEQ ID NO:114, (b) Spacer containing the sequence shown in SEQ ID NO:174, (c) A transmembrane domain containing the sequence shown in SEQ ID NO:138, and (d) Intracellular signaling region including a cytoplasmic signaling region containing the sequence shown in SEQ ID NO:143 and a co-stimulatory signaling region containing the sequence shown in SEQ ID NO:4 A method according to any one of claims 1 to 88, including the method described in any one of claims 1 to 88.

90. The method according to any one of claims 1 to 89, wherein the CAR comprises the sequence shown in SEQ ID NO:

19.

91. Before the aforementioned administration, the subject, 20-40 mg / m² or approximately 20-40 mg / m² 2 The target body surface area can be any 30 or approximately 30 mg / m². 2 Daily administration of fludarabine for 2 to 4 days, and / or 200 to 400 mg / m² or approximately 200 to 400 mg / m² 2 The target body surface area is arbitrarily 300 or approximately 300 mg / m². 2 Lymphocyte depletion therapy, including daily administration of cyclophosphamide for 2 to 4 days. The method according to any one of claims 1 to 90, wherein the patient receives the treatment.

92. In at least one subject within the cohort of subjects with MM, 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%, the specified response or outcome can be achieved at a specified time after the start of administration, optionally. The response or outcome is selected from a group consisting of objective response (OR), complete response (CR), severe complete response (sCR), very good partial response (VGPR), partial response (PR), and minimal response (MR). The response or outcome is OR, or includes OR, and / or The response or outcome is either a complete response (CR) or includes a complete response (CR). The method according to any one of claims 1 to 91.

93. The method according to claim 92, wherein the response or outcome is sustained for more than 3, 6, 9, or 12 months, or for approximately 3, 6, 9, or 12 months.

94. The method according to claim 92 or 93, wherein the response or outcome determined 3, 6, 9, or 12 months after a specified time point, or approximately 3, 6, 9, or 12 months later, is equal to or better than the response or outcome determined at the specified time point.

95. The method according to any one of claims 1 to 94, which 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 a cohort of subjects having MM.

96. The method according to any one of claims 1 to 95, which does not result in severe cytokine release syndrome (CRS) in at least 80%, at least 90%, or at least 95% of subjects in a cohort of subjects having MM.

97. The method according to any one of claims 1 to 96, which 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 a cohort of subjects having MM.

98. The method according to any one of claims 1 to 97, which does not result in severe neurotoxicity in at least 70%, at least 80%, at least 90%, or at least 95% of subjects in a cohort of subjects having MM.

99. The method according to any one of claims 1 to 98, which does not result in severe CRS and severe neurotoxicity in at least 70%, at least 80%, at least 90%, or at least 95% of subjects in a cohort of subjects having MM.

100. The method according to any one of claims 1 to 99, which does not result in severe CRS and severe neurotoxicity in at least 80%, at least 90%, or at least 95% of subjects in a cohort of subjects having MM.

101. The method according to any one of claims 96, 99, and 100, wherein the severe CRS is Grade 3 or higher, Grade 4 or higher, or Grade 5 CRS.

102. The method according to any one of claims 98, 99, and 100, wherein the severe neurotoxicity is grade 3 or higher, grade 4 or higher, or grade 5 CRS.

103. The method according to any one of claims 1 to 102, wherein the composition is administered on an outpatient basis, and / or without hospitalizing the subject, and / or without requiring hospitalization or hospitalization or overnight stay in a hospital, until the subject exhibits or exhibits a persistent fever or a fever that does not decrease by more than 1°C after treatment with an antipyretic.

104. The method according to any one of claims 1 to 103, wherein the composition comprising the manipulated T cells is administered parenterally, and optionally intravenously.

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

106. The method according to any one of claims 1 to 105, wherein a composition containing manipulated T cells is produced by a manufacturing process comprising the following steps: (i) A step of exposing an input composition containing primary T cells, and an input composition containing autologous T cells optionally selected from a subject, to an irritant reagent containing an oligomeric particle reagent containing multiple streptavidin mutein molecules under conditions for stimulating T cells, thereby generating a stimulated population, The oligomer-type particle reagent comprises a first active substance containing an anti-CD3 antibody or its antigen-binding fragment, and a second active substance containing an anti-CD28 antibody or its antigen-binding fragment, in a step, (ii) A step of introducing a heterologous polynucleotide encoding a BCMA-targeting CAR into a stimulated population of T cells, thereby generating a population of transformed cells. (iii) Incubating a population of transformed cells for up to 96 hours, and (iv) A step of collecting T cells from a population of transformed cells, thereby preparing a composition of engineered cells, wherein the collection is performed at a time between 24 and 120 hours, including both values, after the commencement of exposure to the irritant reagent.

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

108. The method according to claim 106 or claim 107, wherein the first active substance and the second active substance each comprise a streptavidin-binding peptide that reversibly binds the first active substance and the second active substance to an oligomeric particle reagent, and optionally the streptavidin-binding peptide comprises an amino acid sequence shown in any one of SEQ ID NO: 266 to 270.

109. The method according to any one of claims 106 to 108, wherein the streptavidin mutein molecule is a tetramer of streptavidin mutein comprising the amino acid residues Val44-Thr45-Ala46-Arg47 or Ile44-Gly45-Ala46-Arg47, and optionally the streptavidin mutein comprises the sequence shown in any one of SEQ ID NO: 257, 272, 275, 277, 279, 273, or 276.

110. The method according to any one of claims 106 to 109, wherein the oligomeric particle reagent comprises 1,000 to 5,000 streptavidin mutein tetramers including values ​​at both ends.

111. The method according to any one of claims 106 to 110, further comprising the step of adding biotin or a biotin analog prior to, after, or during incubation, cell collection.

112. The method according to any one of claims 106 to 111, wherein collection is performed at 48 to 120 hours after initiating exposure to an irritant reagent, including values ​​at both ends.

113. The method according to any one of claims 106 to 112, wherein collection is performed when the integrated vector is detected in the genome, but before the copy number of the integrated vector per diploid genome (iVCN) stabilizes.

114. The method according to any one of claims 106 to 113, wherein the collection is performed before the total number of viable cells at the time of collection becomes more than three times or approximately three times the total number of viable cells in the stimulated population.

115. The method according to any one of claims 106 to 114, wherein the collection is performed when the total number of viable cells at the time of collection is three times or approximately three times, two times or approximately two times, or equal to or approximately equal to the total number of viable cells in the stimulated population.

116. CD27 ＋ CCR7 ＋ The percentage of cells is the total T cells in the transformed cell population, and the total CD3 cells in the transformed cell population. ＋ T cells, total CD4 in a population of transformed cells ＋ All CD8 in a population of T cells or transformed cells ＋ The method according to any one of claims 106 to 115, wherein collection is performed when the number of T cells or their CAR-expressing cells is greater than 50% or approximately greater than 50%.

117. CD45RA ＋ CCR7 ＋ Cells and CD45RA - CCR7 ＋ The percentage of cells is the total T cells in the transformed cell population, and the total CD3 cells in the transformed cell population. ＋ T cells, total CD4 in a population of transformed cells ＋ All CD8 in a population of T cells or transformed cells ＋ The method according to any one of claims 106 to 116, wherein collection is performed when the number of T cells or their CAR-expressing cells is greater than 60% or approximately greater than 60%.

118. (i) comprising engineered CD4+ T cells and engineered CD8+ T cells, (ii) Exhibiting the specified characteristics If a manufacturing process for producing an output composition produces cells in the composition to be administered, and the iteration of the manufacturing process is performed between several different individual subjects, then optionally, multiple output compositions may be produced from a human biological sample, and the predetermined characteristics of the output composition among the multiple output compositions are selected from the following: The average percentage of memory phenotypic cells in the multiple output compositions is approximately 40% to 65%, approximately 40% to 45%, approximately 45% to 50%, approximately 50% to 55%, approximately 55% to 60%, or approximately 60% to 65%; The average percentage of cells exhibiting the central memory phenotype in the multiple output compositions is approximately 40% to 65%, approximately 40% to 45%, approximately 45% to 50%, approximately 50% to 55%, approximately 55% to 60%, or approximately 60% to 65%; The average percentage of cells in the plurality of output compositions that are CD27+, CD28+, CCR7+, CD45RA-, CD45RO+, CD62L+, CD3+, CD95+, granzyme B-, and / or CD127+ is about 40% to about 65%, about 40% to about 45%, about 45% to about 50%, about 50% to about 55%, about 55% to about 60%, or about 60% to about 65%; The average percentage of cells that are CCR7+ / CD45RA- or CCR7+ / CD45RO+ in the multiple output compositions is approximately 40% to approximately 65%, approximately 40% to approximately 45%, approximately 45% to approximately 50%, approximately 50% to approximately 55%, approximately 55% to approximately 60%, or approximately 60% to approximately 65%; In the manipulated CD4+ T cells, optionally CAR+ CD4+ T cells, of the multiple output compositions, the average percentage of central memory CD4+ T cells is approximately 40% to approximately 65%, approximately 40% to approximately 45%, approximately 45% to approximately 50%, approximately 50% to approximately 55%, approximately 55% to approximately 60%, or approximately 60% to approximately 65%; In the manipulated CD8+ T cells, optionally CAR+CD8+ T cells, of the multiple output compositions, the mean percentage of central memory CD8+ T cells is approximately 40% to approximately 65%, approximately 40% to approximately 45%, approximately 45% to approximately 50%, approximately 50% to approximately 55%, approximately 55% to approximately 60%, or approximately 60% to approximately 65%; and / or In the manipulated T cells, optionally CAR+ T cells, of the multiple output compositions, the average percentage of central memory T cells, optionally CD4+ central memory T cells, and CD8+ central memory T cells is approximately 40% to approximately 65%, approximately 40% to approximately 45%, approximately 45% to approximately 50%, approximately 50% to approximately 55%, approximately 55% to approximately 60%, or approximately 60% to approximately 65%. The method according to any one of claims 1 to 117.

119. The composition to be administered is A manufacturing process for producing an output composition exhibiting predetermined characteristics, optionally a threshold number of cells expressing CAR in the output composition, in at least about 80%, about 90%, about 95%, about 97%, about 99%, about 100%, or 100% of human biological samples, performed among multiple different individual subjects. A method according to any one of claims 1 to 118, which is produced by...

120. The method according to any one of claims 1 to 119, wherein MM is relapsed and / or refractory multiple myeloma (r / r MM).

121. A manufactured article comprising a composition containing genetically modified cells expressing a chimeric antigen receptor (CAR) targeting BCMA, and instructions for administering the cell composition according to any one of claims 1 to 120.

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