Methods for administration and treatment of B cell malignancies in adoptive cell therapy - Patents.com

Abstract

Methods of adoptive cell therapy are provided that involve the administration of a dose of cells to treat diseases and conditions, including certain B-cell malignancies. The cells generally express a recombinant receptor, such as a chimeric antigen receptor (CAR). In some embodiments, the methods are for treating subjects with chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL). In some embodiments, the methods are for treating subjects with relapsed or refractory CLL and SLL. Articles of manufacture and prophylactic treatments related to the adoptive therapy methods are also provided. TIFF2022513689000047.tif90128

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. Provisional Patent Application No. 62 / 774,168, entitled "METHODS FOR DOSING AND TREATMENT OF B CELL MALIGNANCIES IN ADOPTIVE CELL THERAPY", filed November 30, 2018, U.S. Provisional Patent Application No. 62 / 774,858, entitled "METHODS FOR DOSING AND TREATMENT OF B CELL MALIGNANCIES IN ADOPTIVE CELL THERAPY", filed December 3, 2018, U.S. Provisional Patent Application No. 62 / 847,897, entitled "METHODS FOR DOSING AND TREATMENT OF B CELL MALIGNANCIES IN ADOPTIVE CELL THERAPY", filed May 14, 2019, and U.S. Provisional Patent Application No. 62 / 847,897, entitled "METHODS FOR DOSING AND TREATMENT OF B CELL MALIGNANCIES IN ADOPTIVE CELL THERAPY", filed May 30, 2019. No. 62 / 854,957, filed Nov. 5, 2019, entitled "METHODS FOR DOSING AND TREATMENT OF B CELL MALIGNANCIES IN ADOPTIVE CELL THERAPY," and U.S. Provisional Application No. 62 / 931,143, filed Nov. 5, 2019, entitled "METHODS FOR DOSING AND TREATMENT OF B CELL MALIGNANCIES IN ADOPTIVE CELL THERAPY," the contents of which are incorporated by reference in their entireties.

[0002] INCORPORATION BY REFERENCE This application is filed together with an electronic sequence listing, which is provided in a file entitled 735042019740SeqList.txt, created on November 21, 2019, and having a size of 34.1 kilobytes. The information in the electronic format of the sequence listing is incorporated by reference in its entirety.

[0003] Field The present disclosure relates in some aspects to adoptive cell therapy, which involves administering a dose of cells to treat subjects with diseases and conditions, such as certain B-cell malignancies, and related methods, compositions, uses, and articles of manufacture. The cells generally express a recombinant receptor, such as a chimeric antigen receptor (CAR). In some embodiments, the disease or condition is chronic lymphocytic leukemia (CLL), such as relapsed or resistant CLL. In some embodiments, the disease or condition is small lymphocytic lymphoma (SLL). In some embodiments, the subject is a particular group or subset of subjects with CLL or SLL, such as heavily pretreated subjects or subjects with poor prognosis. [Background technology]

[0004] background Chronic lymphocytic leukemia (and small lymphocytic lymphoma) are indolent cancers in which immature lymphocytes are found in the blood and bone marrow and / or lymph nodes. Chronic lymphocytic leukemia and small lymphocytic lymphoma are the same disease, but in CLL, the cancer cells are found mostly in the blood and bone marrow. In SLL, the cancer cells are found mostly in the lymph nodes. CLL is considered incurable. Patients eventually relapse or become resistant to available treatments. Although B-cell receptor antagonists have shown improved outcomes for patients with relapsed / resistant CLL, complete remission rates remain low and patients who deteriorate during or after treatment have poor outcomes. Patients with CLL and SLL who have failed B-cell receptor antagonist therapy also need an effective therapy. Methods and uses are provided that meet such needs. Summary of the Invention

[0005] overview Provided herein are methods, uses, compositions, formulations, and articles of manufacture for treating subjects who have or are suspected of having a disease or condition, such as CLL, SLL, or subtypes thereof.The methods and other aspects generally relate to administering T cells, generally engineered T cells, such as those that express or contain recombinant receptors, such as chimeric antigen receptors (CARs) or TCRs, to subjects.

[0006] In some embodiments, the dose of cells or cells administered in connection with any of the provided methods, compositions, articles of manufacture and uses are CD4 + T cells or subtypes or phenotypes thereof (e.g., engineered or recombinant receptor-expressing CD4 + T cells) and / or CD8 + T cells or subtypes thereof (e.g., engineered or recombinant receptor-expressing CD4 + In some embodiments, the CD8 + The cells or subtypes or phenotypes are present in a particular dose or amount or number; in some embodiments, CD4 + The cells or subtypes or phenotypes are present in a particular dose or amount or number. + Cells or their subtypes or phenotypes and CD4 + The cells or subtypes or phenotypes thereof are present in the article or composition or combination or administered in the method in a defined ratio, for example, at or about 1:1, or at or about 1:3 to at or about 3:1. In some embodiments, a dose or administration comprises or is a specific amount or number of one type of population of cells, the ratio being a defined ratio or a ratio that occurs naturally, such as in the blood of the subject from which the cells are derived, or that occurs without selection or control at a specific ratio.

[0007] In some embodiments, CD4 + T cells (or subsets) and CD8 +T cells (or a subset thereof) individually comprise a receptor that specifically binds to a target antigen expressed by the disease or condition, or a cell or tissue thereof, and / or a receptor associated with the disease or condition.

[0008] In some embodiments, the method of treating a subject with chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL) comprises administering to a subject a CD4 + and CD8 + administering to a subject a dose of engineered T cells comprising T cells, + and CD8 + The T cell comprises a chimeric antigen receptor (CAR) that specifically binds to CD19, and the subject relapses after treatment with a Bruton's tyrosine kinase inhibitor (BTKi), venetoclax, or BTKi and venetoclax, becomes resistant to failed treatment, and / or is intolerant to treatment.In certain embodiments, the subject relapses after treatment with a BTKi and venetoclax, becomes resistant to failed treatment, and / or is intolerant to treatment.In some embodiments, the BTKi is ibrutinib.

[0009] In some embodiments, a method of treating a subject with chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL) comprises administering to the subject a dose of engineered T cells comprising CD4+ and CD8+ T cells, wherein the CD4+ and CD8+ T cells comprise a chimeric antigen receptor (CAR) that specifically binds to CD19, and the subject has relapsed following remission after treatment with, has become resistant to failed treatment with, and / or is intolerant to ibrutinib and / or venetoclax. In certain embodiments, the subject has relapsed following remission after treatment with, has become resistant to failed treatment with, and / or is intolerant to treatment with ibrutinib and venetoclax.

[0010] In some embodiments, a dose of engineered T cells is enriched for CD4+ and CD8+ primary human T cells.

[0011] In some embodiments, a dose of engineered T cells comprises a defined ratio of CD4+ cells expressing a CAR:CD8+ cells expressing a CAR, optionally, the ratio is about 1:3 to about 3:1. In some embodiments, a dose of engineered T cells comprises 2.5×10 7 or about 2.5 x 10 7 1.5 x 10 out of 10 total CAR-expressing cells 8 or about 1.5 x 10 8 Contains total CAR-expressing cells.

[0012] In some embodiments, the administering comprises administering a plurality of separate compositions, the plurality of separate compositions comprising CD4 + T cells and CD8 + A first composition comprising one of CD4 T cells and + T cells and CD8 + and a second composition comprising another of the CD4 T cells. + Receptors contained by T cells and / or CD8 + The receptors carried by the T cells include T cells with recombinant receptors and / or CD4 + T cells and / or CD8 + The T cells are genetically engineered to express the receptor.

[0013] In some aspects, a method includes treating a subject having or suspected of having chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL), the method includes administering to the subject a dose of engineered T cells comprising CD4+ and CD8+ T cells, the CD4+ and CD8+ T cells comprising a CAR that specifically binds to CD19, the administering includes administering a plurality of separate compositions, the plurality of separate compositions including a first composition comprising one of the CD4+ T cells and the CD8+ T cells, and a second composition comprising the other of the CD4+ T cells and the CD8+ T cells.

[0014] In some embodiments, a dose of engineered CD4 + and CD8 + T cells are fused to CD4 T cells expressing CAR at a defined ratio of approximately 1:3 to approximately 3:1. + Cells: CAR-expressing CD8 + In some embodiments, a dose of engineered T cells comprises 2.5×10 7 or about 2.5 x 10 7 1.5 x 10 out of 10 total CAR-expressing cells 8 or about 1.5 x 10 8 In some embodiments, a dose of engineered T cells is enriched for CD4+ and CD8+ primary human T cells.

[0015] In some embodiments, a dose of cells enriched for engineered CD4+ and CD8+ T cells comprises greater than or about 70%, greater than or about 75%, greater than or about 80%, greater than or about 85%, greater than or about 85%, greater than or about 90%, greater than or about 95%, or greater than or about 98% CD4+ and CD8+ primary human T cells.

[0016] In some embodiments, a dose of engineered CD4 + and CD8 + T cells are fused to CD4 T cells expressing CAR at a defined ratio of 1:1 or approximately 1:1. + Cells: CAR-expressing CD8 + In some embodiments, a dose of engineered T cells comprises 2.5×10 7 or about 2.5 x 10 7 1.0 x 10 out of 10 total CAR-expressing cells 8 or about 1.0 x 10 8 In some embodiments, a dose of engineered T cells comprises 2.5×10 total CAR-expressing cells. 7 or about 2.5 x 10 7In some embodiments, a dose of engineered T cells comprises 5×10 total CAR-expressing cells. 7 or about 5 x 10 7 In some embodiments, the engineered T cells comprise 1×10 total cells or total CAR-expressing cells. 8 or about 1 x 10 8 Total cells or total CAR-expressing cells.

[0017] In some embodiments, CD4 + CAR and / or CD8 contained by T cells + The CAR contained by the T cell may contain the same CAR and / or may be CD4 + T cells and / or CD8 + The T cells are genetically engineered to express the same CAR.

[0018] In certain embodiments, the first composition is CD8 + In some embodiments, the first composition comprises CD4+ T cells and the second composition comprises CD4+ T cells. In some embodiments, the initiation of administration of the first composition occurs before the initiation of administration of the second composition.

[0019] In some embodiments, the administration of the first composition and the administration of the second composition occur on the same day and are spaced 36 hours or less apart, 24 hours or less apart, 12 hours or less apart, 6 hours or less apart, 4 hours or less apart, 2 hours or less apart, or 1 hour or less apart, or 30 minutes or less apart. In some embodiments, the administration of the first composition and the administration of the second composition occur between 0 or about 0 and 48 or about 48 hours, between 0 or about 0 and 36 or about 36 hours, between 0 or about 0 and 24 or about 24 hours, between 0 or about 0 and 12 or about 12 hours, between 0 or about 0 and 6 or about 6 hours, between 0 or about 0 and 2 or about 2 hours, between 0 or about 0 and 1 or about 1 hour, between 0 or about 0 and 30 or about 1 hour, or ... is between about 30 minutes, between 30 or about 30 minutes and 48 or about 48 hours, between 30 or about 30 minutes and 36 or about 36 hours, between 30 or about 30 minutes and 24 or about 24 hours, between 30 or about 30 minutes and 12 or about 12 hours, between 30 or about 30 minutes and 6 or about 6 hours, between 30 or about 30 minutes and 4 or about 4 hours, between 30 or about 30 minutes and 2 or about 2 hours, between 30 or about 30 minutes and 1 or within about 1 hour, between 1 or about 1 hour and 48 or about 48 hours, between 1 or about 1 hour and 36 or about 36 hours, between 1 or about 1 hour and 24 or about 24 hours, between 1 or about 1 hour and 12 or about 12 hours, between 1 or about 1 hour and 6 or about 6 hours, between 1 or about 1 hour and 4 or about 4 hours, between 1 or about 1 hour and 2 or about 2 hours, between 2 or about 2 hours and 48 or about 48 hours between 2 or about 2 hours and 36 or about 36 hours; between 2 or about 2 hours and 24 or about 24 hours; between 2 or about 2 hours and 12 or about 12 hours; between 2 or about 2 hours and 6 or about 6 hours; between 2 or about 2 hours and 4 or about 4 hours; between 4 or about 4 hours and 48 or about 48 hours; between 4 or about 4 hours and 36 or about 36 hours; between 4 or about 4 hours and 24 or about 24 hours;Between 4 or about 4 hours and 12 or about 12 hours, between 4 or about 4 hours and 6 or about 6 hours, between 6 or about 6 hours and 48 or about 48 hours, between 6 or about 6 hours and 36 or about 36 hours, between 6 or about 6 hours and 24 or about 24 hours, between 6 or about 6 hours and 12 or about 12 hours, between 12 or about 12 hours and 48 or about 48 hours, between 12 or about 12 hours and 36 or about 36 hours, between 12 or about 12 hours and 24 or about 24 hours, between 24 or about 24 hours and 48 or about 48 hours, between 24 or about 24 hours and 36 or about 36 hours, or between 36 or about 36 hours and 48 or about 48 hours.

[0020] In some embodiments, the administration of the first composition and the administration of the second composition occur on the same day, and occur about 0 to about 12 hours apart, about 0 to about 6 hours apart, or about 0 to 2 hours apart; or the start of administration of the first composition and the start of administration of the second composition occur about 1 minute to about 1 hour apart, or about 5 minutes to about 30 minutes apart. In some embodiments, the first composition and the second composition are administered 2 hours or less, 1 hour or less, 30 minutes or less, 15 minutes or less, 10 minutes or less, or 5 minutes or less apart.

[0021] In some embodiments, the first composition and the second composition are administered no more than 2 hours, no more than 1 hour, no more than 30 minutes, no more than 15 minutes, no more than 10 minutes, or no more than 5 minutes apart.

[0022] In some embodiments of any of the provided methods, the subject has or is suspected of having CLL; or the subject is identified or selected as having CLL. In some embodiments, the CLL is relapsed or refractory CLL.

[0023] In some embodiments, the subject has or is suspected of having SLL; or the subject is identified or selected as having SLL. In some embodiments, the SLL is recurrent or refractory SLL.

[0024] In some embodiments, prior to administration of a dose of engineered T cells, the subject has been treated with one or more prior therapies for CLL or SLL other than another dose of cells expressing a CAR or lymphodepleting therapy. In some embodiments, the one or more prior therapies include at least two prior therapies, optionally, three, four, five, six, seven, eight, nine or more prior therapies.

[0025] In some embodiments, at the time of administration of the dose of cells or immediately before administration, the subject has relapsed following remission after treatment with one or more previous therapies for CLL or SLL, or has become resistant to the treatment, has failed the treatment, and / or has been intolerant to the treatment. In some embodiments, the subject has relapsed following remission after treatment with two or more previous therapies, or has become resistant to the treatment, has failed the treatment, and / or has been intolerant to the treatment. In some embodiments, at the time of administration of the dose of cells or immediately before administration, the subject has relapsed following remission after treatment with three or more previous therapies, or has become resistant to the treatment, has failed the treatment, and / or has been intolerant to the treatment. In some embodiments, the previous therapy is selected from a kinase inhibitor, optionally an inhibitor of Bruton's tyrosine kinase (BTK), optionally ibrutinib; venetoclax; combination therapy including fludarabine and rituximab; radiation therapy; and hematopoietic stem cell transplantation (HSCT). In some embodiments, the prior therapy includes ibrutinib and / or venetoclax. In some embodiments, the prior therapy includes ibrutinib and venetoclax.

[0026] In some embodiments, the subject has relapsed following remission after treatment with ibrutinib and / or venetoclax, has become resistant to failed treatment with them, and / or is intolerant to them. In some embodiments, the subject has relapsed following remission after treatment with ibrutinib and venetoclax, has become resistant to failed treatment with them, and / or is intolerant to them.

[0027] In some embodiments, at or before administration of a dose of cells, The target is: Optionally, one or more cytogenetic abnormalities associated with high-risk CLL selected from among complex karyotype or cytogenetic abnormalities, del 17p, unmutated IGVH gene, and TP53 mutations Identified or identified as having; The subject is identified or has been identified as having high-risk CLL.

[0028] In some embodiments, the subject is identified or has been identified as having an ECOG status of 0 or 1; and / or the subject does not have an ECOG status of > 1. In some embodiments, the subject does not have Richter's transformation of CLL or SLL at or shortly before administration of a dose of engineered cells or at or shortly before administration of lymphodepletion therapy.

[0029] In some embodiments, the subject is an adult and / or is greater than or greater than about 50, 60, or 70 years of age.

[0030] In some embodiments, the T cells are primary T cells obtained from a subject. In some embodiments, the T cells are autologous to the subject. In some embodiments, a dose of engineered cells are live cells.

[0031] In some embodiments, lymphodepletion therapy is administered to the subject prior to administration of a dose of engineered cells. In some embodiments, the subject is preconditioned with lymphodepletion therapy. In some embodiments, the lymphodepletion therapy includes administration of fludarabine and / or cyclophosphamide. In some embodiments, the lymphodepletion therapy includes administration of about 200-400 mg / m 2 (inclusive), optionally 300 or about 300 mg / m 2 of cyclophosphamide, and / or approximately 20-40 mg / m 2 , optionally 30 mg / m 2 of fludarabine daily for 2-4 days, optionally 3 days. In some embodiments, lymphodepletion therapy comprises administration of at or about 300 mg / m 2 of cyclophosphamide and approximately 30 mg / m 2 of fludarabine daily for three days, optionally wherein the dose of cells is administered at least or at least about 2-7 days after lymphodepletion therapy or at least or at least about 2-7 days after the initiation of lymphodepletion therapy.

[0032] In some embodiments, at or before administration of a dose of cells, The subject may have one or more cytogenetic abnormalities associated with high-risk CLL or SLL, optionally selected from among complex karyotype or cytogenetic abnormalities, del 17p, unmutated IGVH gene, and TP53 mutations. and / or The subject is identified or has been identified as having high-risk CLL or SLL. In some embodiments, at or immediately prior to administration of a dose of cells, the subject has relapsed following remission after treatment with two or more previous therapies, or has become resistant to the treatment, has failed the treatment, and / or has been intolerant to the treatment. In some embodiments, at or immediately prior to administration of a dose of cells, the subject is identified or identified as having standard risk CLL or SLL. In some embodiments, at or immediately prior to administration of a dose of cells, the subject has relapsed following remission after treatment with three or more previous therapies, or has become resistant to the treatment, has failed the treatment, and / or has been intolerant to the treatment. In some embodiments, at or immediately prior to administration of a dose of cells, the subject is identified or identified as being intolerant to an inhibitor of Bruton's tyrosine kinase (BTK) and has received an inhibitor of BTK for less than or less than about six months, and / or is ineligible for treatment with an inhibitor of BTK.

[0033] In some embodiments, (i) the subject is identified or has been identified as having high-risk CLL or SLL, and at the time of or immediately prior to administration of a dose of the cells, the subject has relapsed following remission after treatment with one or more prior therapies other than an inhibitor of BTK, or has become resistant to, failed, and / or been intolerant to the treatment; or (ii) the subject is identified or has been identified as having standard-risk CLL or SLL, and at the time of or immediately prior to administration of a dose of the cells, the subject has relapsed following remission after treatment with two or more prior therapies other than an inhibitor of BTK, or has become resistant to, failed, and / or been intolerant to the treatment.

[0034] In some embodiments, administration of the cell dose and / or administration of the lymphodepletion therapy is via outpatient delivery, hi some embodiments, a dose of cells is administered parenterally, optionally intravenously.

[0035] In some embodiments, the response in at least 35%, at least 40%, at least 50%, at least 60%, or at least 70% of treated subjects is a complete response (CR) in subjects treated by the method. In some embodiments, the duration of response before progression lasts for more than 3 months or more than 6 months. In some embodiments, more than 50%, more than 60%, or more than 70% of subjects treated by the method have undetectable minimal residual disease (MRD) for at least 1 month, at least 2 months, at least 3 months, or at least 6 months after administration of a dose of cells.

[0036] In some embodiments, 10% or less of the subjects treated by the method exhibit cytokine release syndrome (CRS) greater than Grade 2. In some embodiments, 10% or less, 20% or less, 30% or less, or 40% or less of the subjects treated by the method exhibit neurotoxicity greater than Grade 2. In some embodiments, the subjects treated by the method include subjects who have relapsed following remission following treatment with, become resistant to, failed treatment with, and / or are intolerant to ibrutinib and venetoclax.

[0037] In some aspects, the method includes: assaying a biological sample for a level, amount, or concentration of TNF-α, wherein the biological sample is optionally derived from a subject who is a candidate for treatment with a cell therapy, and the cell therapy comprises a dose of engineered cells, including T cells expressing a CAR for treating a disease or condition, and wherein the biological sample was obtained from the subject prior to administering the cell therapy and / or the biological sample does not comprise a CAR and / or the engineered cells; and Comparing the level, amount, or concentration of TNF-α to a threshold level, (1) identifying the subject as at risk for developing grade 3 or higher neurotoxicity after administration of the cell therapy if the level, amount, or concentration of TNF-α is at or above the threshold level; and (2) identifying the subject as not at risk for developing grade 3 or higher neurotoxicity after administration of the cell therapy if the level, amount, or concentration of TNF-α is below the threshold level.

[0038] In some embodiments, the subject is identified as being at risk for developing grade 3 or higher neurotoxicity, and the method further includes: (i) administering cell therapy to the subject, optionally at a reduced dose, where optionally (a) the method further includes administering to the subject an agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset or risk of onset of neurotoxicity, and / or (b) administration of cell therapy to the subject is performed or designated to be performed in an inpatient setting and / or with one or more days of hospitalization; or (ii) administering to the subject an alternative treatment other than cell therapy to treat the disease or condition.

[0039] In some embodiments, a subject is identified as not at risk for developing Grade 3 or higher neurotoxicity following administration of the cell therapy, and (i) optionally, the subject is not administered an agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset of, or the risk of onset of, toxicity, unless or until the subject exhibits signs or symptoms of toxicity, at or after the subject exhibits persistent fever or fever that has or has not or does not decrease by more than 1° C. following treatment with an antipyretic; and / or (ii) optionally, unless or until the subject exhibits persistent fever or fever that has or has not or does not decrease by more than 1° C. following treatment with an antipyretic, the administration and all follow-up is performed on an outpatient basis and / or without admitting the subject to a hospital and / or without an overnight stay in a hospital and / or without the need for hospitalization or overnight stay in a hospital.

[0040] In some embodiments, the assaying step includes: (a) contacting the biological sample with one or more reagents capable of detecting TNF-α or specific for TNF-α, optionally wherein the one or more reagents include an antibody that specifically recognizes TNF-α; and (b) detecting the presence or absence of a complex comprising the one or more reagents and TNF-α.

[0041] In some aspects, the method includes administering to the subject a cell therapy for treating a disease or condition, the cell therapy comprising a dose of engineered cells comprising T cells expressing a CAR, wherein (1) if the subject has a level, amount, or concentration of TNF-α in a biological sample from the subject that is at or above a threshold level, the subject is identified as at risk for developing grade 3 or higher neurotoxicity following administration of the cell therapy, and the method includes: (i) administering the cell therapy at a reduced dose to the subject, (ii) further administering to the subject an agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset or risk of onset of toxicity; and / or (iii) administering to the subject the cell therapy is administered or specified to be administered in an inpatient setting and / or with one or more days of hospitalization; or (2) if the subject is selected or identified as having a level, amount, or concentration of TNF-α in a biological sample from the subject that is below a threshold level, the subject is identified as at risk for developing grade 3 or higher neurotoxicity following administration of the cell therapy. and (i) optionally, at or after the subject exhibits persistent fever or a fever that has or has not or will not decrease by more than 1° C. after treatment with an antipyretic agent, said administration and all follow-up is performed on an outpatient basis and / or without the subject being admitted to a hospital and / or without an overnight stay in a hospital and / or without the need for hospitalization or overnight stay in a hospital, unless or until the subject exhibits persistent fever or a fever that has or has not or will not decrease by more than 1° C. after treatment with an antipyretic agent, said administration and all follow-up is performed on an outpatient basis and / or without the subject being admitted to a hospital and / or without an overnight stay in a hospital, unless or until the subject exhibits persistent fever or a fever that has or has not or will not decrease by more than 1° C. after treatment with an antipyretic agent, said administration and all follow-up is performed on an outpatient basis and / or without the subject being admitted to a hospital and / or without an overnight stay in a hospital, and / or without the need for hospitalization or overnight stay in a hospital, and the subject is a candidate for treatment with a cell therapy, and said biological sample and / or said biological sample obtained from the subject prior to administration of the cell therapy does not contain a CAR and / or said engineered cells.

[0042] In some embodiments, the subject is identified as at risk for developing grade 3 or greater neurotoxicity following administration of the cell therapy, and the method includes administering an agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset or risk of onset of the toxicity, wherein the agent is administered to the subject simultaneously with the cell therapy or within 3 days of administration of the cell therapy to the subject.

[0043] In some embodiments, the threshold level is within 25%, 20%, 15%, 10%, or 5% above the median or mean of a level, amount, or concentration of TNF-α in biological samples obtained from a group of subjects prior to receiving cell therapy, and / or is within a standard deviation above said median or mean, or is the median or mean of said level, amount, or concentration, or is about the median or mean of said level, amount, or concentration, and each of the subjects in the group does not exhibit any grade of neurotoxicity following administration of a dose of engineered cells expressing a CAR to treat the same disease or condition; the threshold level is within 25%, 20%, 15%, 10%, or 5% above the median or mean of a level, amount, or concentration of TNF-α in biological samples obtained from a group of subjects prior to receiving cell therapy, and / or is within a standard deviation above said median or mean, or is about the median or mean of said level, amount, or concentration, and each of the subjects in the group does not exhibit any grade of neurotoxicity following administration of a dose of engineered cells expressing a CAR to treat the same disease or condition; or greater than 1.25-fold, 1.3-fold, 1.4-fold, or 1.4-fold higher than the median or mean level, amount, or concentration of TNF-α in the samples, and each of the subjects in the group does not exhibit any grade of neurotoxicity following administration of a dose of engineered cells expressing a CAR to treat the same disease or condition; the threshold level is 1.25-fold or greater than 1.25-fold, 1.3-fold or greater than 1.3-fold, 1.4-fold or greater than 1.4-fold, or 1.5-fold or greater than 1.5-fold higher than the level, amount, or concentration of TNF-α in biological samples obtained from a group of normal or healthy subjects who are not candidates for cell therapy treatment.

[0044] In some embodiments, the threshold level is at or above 1000 pg / mL of the biological sample, at or above 1100 pg / mL of the biological sample, at or above 1200 pg / mL of the biological sample, at or above 1300 pg / mL of the biological sample, at or above 1400 pg / mL of the biological sample, or at or above 1500 pg / mL of the biological sample.

[0045] In some aspects, the method includes: (a) assaying a biological sample from a subject receiving a cell therapy comprising a dose of engineered cells comprising T cells expressing a CAR to treat a disease or condition for a level, amount, or concentration of IL-16, wherein the biological sample is obtained from the subject within 1, 2, or 3 days after initiation of administration of the cell therapy; and (b) comparing the level, amount, or concentration of IL-16 to a threshold level, wherein (1) if the level, amount, or concentration of IL-16 is at or above the threshold level, the subject is identified as at risk for developing grade 3 or higher neurotoxicity; and (2) if the level, amount, or concentration of IL-16 is below the threshold level, the subject is identified as not at risk for developing grade 3 or higher neurotoxicity.

[0046] In some embodiments, if a subject is identified as being at risk for developing grade 3 or higher neurotoxicity, an agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset or risk of onset of toxicity is administered.

[0047] In some embodiments, the assaying step comprises: (a) contacting the biological sample with one or more reagents capable of detecting or specific for IL-16, optionally wherein the one or more reagents comprise an antibody that specifically recognizes IL-16; (b) detecting the presence or absence of a complex comprising the reagent and IL-16. In some embodiments, the method further comprises administering a cell therapy to the subject prior to the assaying step.

[0048] In some aspects, the method includes administering to a subject identified as at risk for developing grade 3 or higher neurotoxicity an agent or other treatment capable of treating, preventing, delaying, reducing, or attenuating the onset or risk of onset of toxicity, wherein the subject has previously received a cell therapy to treat a disease or condition, and the subject is selected or identified as at risk for developing grade 3 or higher neurotoxicity at or shortly before administration of the agent if the level or amount or concentration of IL-16 in a biological sample obtained from the subject within 1, 2, or 3 days of initiating administration of the cell therapy is above a threshold level.

[0049] In some embodiments, the step of administering the agent occurs when the subject exhibits persistent fever or a fever that has or has not decreased by more than 1° C. following treatment with an antipyretic. In some embodiments, administration of the cell therapy to the subject occurs on an outpatient basis, and if the level, amount, or concentration of IL-16 is above a threshold level, the method includes hospitalizing the patient in a hospital for one or more days.

[0050] In some embodiments, the threshold level is within 25%, 20%, 15%, 10%, or 5% above the median or mean value of a level, amount, or concentration of IL-16 in a biological sample obtained from a group of subjects within 1, 2, or 3 days after receiving a cell therapy comprising administration of a dose of engineered cells expressing a CAR to treat the same disease or condition, and / or is within a standard deviation above said median or mean value, or is at or about said median or mean value of said level, amount, or concentration, wherein each of the subjects in the group did not exhibit any grade of neurotoxicity following administration of the cell therapy.

[0051] In some embodiments, the threshold level is 1.3-fold or greater, 1.4-fold or greater, 1.5-fold or greater, 1.6-fold or greater, 1.7-fold or greater, 1.8-fold or greater, 1.9-fold or greater, or 2.0-fold or greater than 2.0-fold greater than the median or mean level, amount, or concentration of IL-16 in biological samples obtained from a group of subjects prior to receiving cell therapy, where each of the subjects in the group did not exhibit any grade of neurotoxicity following administration of a dose of engineered cells expressing a CAR to treat the same disease or condition.

[0052] In some embodiments, the threshold level is 1.3-fold or greater, 1.4-fold or greater, 1.5-fold or greater, 1.6-fold or greater, 1.7-fold or greater, 1.8-fold or greater, 1.9-fold or greater, or 2.0-fold or greater than 2.0-fold higher than the level, amount, or concentration of IL-16 in a biological sample obtained from a group of normal or healthy subjects who are not candidates for cell therapy treatment.

[0053] In some embodiments, the threshold level is at or above 1000 pg / mL of the biological sample, at or above 1500 pg / mL of the biological sample, at or above 2000 pg / mL of the biological sample, at or above 2500 pg / mL of the biological sample, or at or above 3000 pg / mL of the biological sample.

[0054] In some aspects, the biological sample is or is obtained from a blood, plasma, or serum sample. In some aspects, the biological sample is a serum sample. In some aspects, assaying or evaluating the cellular analyte comprises an immunoassay. In some aspects, the disease or condition is cancer. In some aspects, the disease or condition is myeloma, leukemia, or lymphoma.

[0055] In some embodiments, the antigen is ROR1, B cell maturation antigen (BCMA), carbonic anhydrase 9 (CAIX), tEGFR, Her2 / neu (receptor tyrosine kinase erbB2), L1-CAM, CD19, CD20, CD22, mesothelin, CEA, and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimer, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kinase insert domain receptor (kdr), kappa light chain, Lewis Y, L1 cell adhesion molecule, (L1-CAM), melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, preferentially expressed antigen in melanoma (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250 / CAIX, HLA-AI MAGE Al, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7 / 8, avb6 integrin, 8H9, NCAM, VEGF receptor, 5T4, fetal AchR, NKG2D ligand, CD44v6, dual antigen, cancer testis antigen, mesothelin, mouse CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, oncofetal antigen, ROR1, TA G72, VEGF-R2, carcinoembryonic antigen (CEA), Her2 / neu, estrogen receptor, progesterone receptor, ephrin B2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms' tumor 1 (WT-1), cyclin, cyclin A2, CCL-1, CD138, G protein-coupled receptor 5D (GPCR5D), or pathogen-specific antigen.

[0056] In some embodiments, the antigen is CD19.

[0057] In some embodiments, the disease or condition is a B-cell malignancy and / or is acute lymphoblastic leukemia (ALL), adult ALL, chronic lymphoblastic leukemia (CLL), small lymphocytic lymphoma (SLL), non-Hodgkin's lymphoma (NHL), and diffuse large B-cell lymphoma (DLBCL). In some embodiments, the disease or condition is CLL or SLL.

[0058] In some aspects, the agent is or includes tocilizumab, siltuximab, or dexamethasone.

[0059] In some embodiments, the CAR comprises an extracellular antigen-binding domain specific for CD19, a transmembrane domain, a cytoplasmic signaling domain derived from a costimulatory molecule, optionally 4-1BB, and a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule, optionally CD3ζ; the CAR, in turn, comprises an extracellular antigen-binding domain specific for CD19, a transmembrane domain, a cytoplasmic signaling domain derived from a costimulatory molecule, and a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule.

[0060] In some embodiments, the antigen binding domain is an scFv.

[0061] In some embodiments, the scFv comprises a CDRL1 sequence of RASQDISKYLN (SEQ ID NO: 35), a CDRL2 sequence of SRLHSGV (SEQ ID NO: 36), and / or a CDRL3 sequence of GNTLPYTFG (SEQ ID NO: 37), and / or a CDRH1 sequence of DYGVS (SEQ ID NO: 38), a CDRH2 sequence of VIWGSETTYYNSALKS (SEQ ID NO: 39), and / or a CDRL3 sequence of YAMDYWG (SEQ ID NO: the scFv comprises the variable heavy chain region of FMC63 and the variable light chain region of FMC63, and / or the CDRL1 sequence of FMC63, the CDRL2 sequence of FMC63, the CDRL3 sequence of FMC63, the CDRH1 sequence of FMC63, the CDRH2 sequence of FMC63, and the CDRH3 sequence of FMC63, or binds to the same epitope as or competes for binding with any of the foregoing; the scFv comprises a VH set forth in SEQ ID NO:41 and a VL set forth in SEQ ID NO:42, optionally wherein the VH and VL are separated by a flexible linker, optionally wherein the flexible linker is or comprises the sequence set forth in SEQ ID NO:24; and / or the scFv is or comprises the sequence set forth in SEQ ID NO:43.

[0062] In some embodiments, the costimulatory signaling region is the signaling domain of CD28 or 4-1BB. In some embodiments, the costimulatory signaling region is the signaling domain of 4-1BB.

[0063] In some embodiments, the costimulatory domain comprises SEQ ID NO: 12, or a variant thereof having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto. In some embodiments, the primary signaling domain is a CD3 zeta signaling domain. In some embodiments, the primary signaling domain comprises SEQ ID NO: 13 or 14 or 15, having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto.

[0064] In some embodiments, the CAR further comprises a spacer between the transmembrane domain and the scFv. In some embodiments, the spacer is a polypeptide spacer that includes or consists of all or a portion of an immunoglobulin hinge or a modified version thereof, optionally an IgG4 hinge or a modified version thereof. In some embodiments, the spacer is about 15 amino acids or less and does not include the CD28 extracellular region and the CD8 extracellular region. In some embodiments, the spacer is 12 amino acids in length or about 12 amino acids in length.

[0065] In some embodiments, the spacer has or consists of a sequence encoded by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, or a variant of any of the foregoing having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto; and / or X 1 is glycine, cysteine, or arginine and X 2 is cysteine ​​or threonine; 1 PPX 2It comprises or consists of P.

[0066] In some of any such embodiments, the subject is a human subject.

[0067] In some of the provided articles of manufacture, the article of manufacture includes a cell therapy composition or one of a plurality of cell therapy compositions comprising T cells expressing an anti-CD19 chimeric antigen receptor (CAR) and instructions for administering the cell therapy, the instructions specifying the step of administering the T cell composition according to any of the provided methods. [The present invention 1001] 1. A method of treating a subject with chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL), comprising administering to a subject a CD4 + and CD8 + administering to a subject a dose of engineered T cells comprising T cells, + and CD8 + wherein the T cells comprise a chimeric antigen receptor (CAR) that specifically binds to CD19, and the dose of engineered T cells (i) enriched for CD4+ and CD8+ primary human T cells; (ii) a defined ratio of CD4+ cells expressing a CAR:CD8+ cells expressing a CAR, optionally wherein the ratio is from about 1:3 to about 3:1; and (iii) 2.5 × 10 7 or about 2.5 x 10 7 1.5 x 10 out of 10 total CAR-expressing cells 8 or about 1.5 x 10 8 containing total CAR-expressing cells, The method. [The present invention 1002] 1. A method of treating a subject with chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL), comprising administering to a subject a CD4 + and CD8 + administering to a subject a dose of engineered T cells comprising T cells, + and CD8 + The method, wherein the T cells comprise a chimeric antigen receptor (CAR) that specifically binds CD19, and the subject has relapsed following remission after treatment with, has become resistant to failed treatment with, and / or is intolerant to, a Bruton's tyrosine kinase inhibitor (BTKi) and venetoclax. [The present invention 1003] The method of claim 1002, wherein the BTKi is ibrutinib. [The present invention 1004] 1. A method of treating a subject with chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL), comprising administering to a subject a CD4 + and CD8 + administering to a subject a dose of engineered T cells comprising T cells, + and CD8 + The method, wherein the T cells comprise a chimeric antigen receptor (CAR) that specifically binds to CD19, and the subject has relapsed following remission after treatment with, has become resistant to failed treatment with, and / or is intolerant to ibrutinib and venetoclax. [The present invention 1005] The method of any of claims 1002 to 1004, wherein said dose of engineered T cells is enriched for CD4+ and CD8+ primary human T cells. [The present invention 1006] Any of the methods of claims 1002-1005, wherein said dose of engineered T cells comprises a defined ratio of CD4+ cells expressing a CAR:CD8+ cells expressing a CAR, optionally wherein the ratio is from about 1:3 to about 3:1. [The present invention 1007] The administering comprises administering a plurality of separate compositions, the plurality of separate compositions comprising CD4 + T cells and CD8 + A first composition comprising one of CD4 T cells and + T cells and CD8 + and a second composition comprising the other of the T cells. [The present invention 1008] A method of treating a subject having or suspected of having chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL), comprising administering to the subject a CD4 + and CD8 + administering to a subject a dose of engineered T cells comprising T cells, + and CD8 + The T cells comprise a CAR that specifically binds to CD19, and the administering comprises administering a plurality of separate compositions, the plurality of separate compositions comprising a CAR that specifically binds to CD4 + T cells and CD8 + A first composition comprising one of T cells and CD4 + T cells and CD8 + and a second composition comprising another of the T cells. [The present invention 1009] The dose of engineered T cells comprises a defined ratio of about 1:3 to about 3:1 of CD4 T cells expressing a CAR. + Cells: CAR-expressing CD8 + The method of any one of claims 1001 to 1008, comprising a cell. [The present invention 1010] The dose of engineered T cells is 2.5×10 7 or about 2.5 x 10 7 1.5 x 10 out of 10 total CAR-expressing cells 8 or about 1.5 x 10 8 Any of the methods of 1002 to 1009, comprising a total of CAR-expressing cells. [The present invention 1011] The method of any of claims 1008 to 1010, wherein said dose of engineered T cells is enriched for CD4+ and CD8+ primary human T cells. [The present invention 1012] Any of the methods of claims 1001-1011, wherein said dose of engineered T cells enriched for primary human CD4+ and CD8+ T cells comprises greater than or about 70%, greater than or about 75%, greater than or about 80%, greater than or about 85%, greater than or about 90%, greater than or about 95%, or greater than or about 98% CD4+ and CD8+ primary human T cells. [The present invention 1013] The dose of engineered T cells comprises a defined ratio of 1:1 or approximately 1:1 CD4 + Cells: CAR-expressing CD8 + The method of any one of claims 1001 to 1012, comprising a cell. [The present invention 1014] The dose of engineered T cells is 2.5×10 7 or about 2.5 x 10 7 1.0 x 10 out of 10 total CAR-expressing cells 8 or about 1.0 x 10 8 Any of the methods of claims 1001 to 1013, comprising a total of CAR-expressing cells. [The present invention 1015] The dose of engineered T cells is 2.5×10 7 or about 2.5 x 10 7 Any of the methods of claims 1001 to 1014, comprising a total of CAR-expressing cells. [The present invention 1016] The dose of engineered T cells is 5×10 7 or about 5 x 10 7 The method of any of claims 1001 to 1014, comprising a total number of cells or a total number of CAR-expressing cells. [The present invention 1017] The dose of engineered T cells is 1×10 8 or about 1 x 10 8 The method of any of claims 1001 to 1014, comprising a total number of cells or a total number of CAR-expressing cells. [The present invention 1018] CD4 + CAR and / or CD8 contained by T cells + The CAR contained by the T cell contains the same CAR and / or is a CD4 + T cells and / or CD8 + The method of any of claims 1001 to 1017, wherein the T cells are genetically engineered to express the same CAR. [The present invention 1019] The first composition is a CD8 + The method of any of claims 1007 to 1018, wherein the first composition comprises CD4+ T cells and the second composition comprises CD4+ T cells. [The present invention 1020] The method of claim 1019, wherein initiation of administration of the first composition occurs before initiation of administration of the second composition. [The present invention 1021] The method according to any one of claims 1007 to 1020, wherein the administration of the first composition and the administration of the second composition are performed at an interval of 48 hours or less. [The present invention 1022] Any of the methods of claims 1007 to 1021, wherein the administration of the first composition and the administration of the second composition are performed at intervals of 36 hours or less, 24 hours or less, 12 hours or less, 6 hours or less, 4 hours or less, 2 hours or less, 1 hour or less, or 30 minutes or less. [The present invention 1023] the administration of the first composition and the administration of the second composition occur on the same day and occur about 0 to about 12 hours apart, about 0 to about 6 hours apart, or about 0 to about 2 hours apart; or The initiation of administration of the first composition and the initiation of administration of the second composition are performed at intervals of about 1 minute to about 1 hour or about 5 minutes to about 30 minutes. Any of the methods of 1007 to 1022 of the present invention. [The present invention 1024] The method of any of claims 1007 to 1023, wherein the first composition and the second composition are administered at an interval of 2 hours or less, 1 hour or less, 30 minutes or less, 15 minutes or less, 10 minutes or less, or 5 minutes or less. [The present invention 1025] the subject has or is suspected of having CLL; or The subject is identified or selected as having CLL. Any of the methods according to the present invention 1001 to 1024. [The present invention 1026] The method of any of claims 1001 to 1025, wherein the CLL is relapsed or resistant CLL. [The present invention 1027] the subject has or is suspected of having SLL; or The subject is identified or selected as having SLL, Any of the methods of 1001 to 1026 of the present invention. [The present invention 1028] Any of the methods of 1001 to 1024 and 1027, wherein the SLL is recurrent or resistant SLL. [The present invention 1029] Any of the methods of claims 1001-1028, wherein prior to administration of said dose of engineered T cells, the subject has been treated with one or more previous therapies for CLL or SLL other than another dose of cells expressing a CAR or lymphodepleting therapy. [The present invention 1030] The method of claim 1029, wherein the one or more prior therapies includes at least two prior therapies, optionally three, four, five, six, seven, eight, nine or more prior therapies. [The present invention 1031] Any of the methods of claims 1001-1030, wherein at the time of administration of said dose of engineered T cells or immediately prior to administration, the subject has relapsed following remission after treatment with one or more prior therapies for CLL or SLL, or has become resistant to said treatment, failed said treatment, and / or was intolerant to said treatment. [The present invention 1032] Any of the methods of claims 1001 to 1031, wherein at the time of administration of said dose of engineered T cells or immediately prior to administration, the subject has relapsed following remission after treatment with two or more prior therapies or has become resistant to said treatment, failed said treatment, and / or was intolerant to said treatment. [The present invention 1033] Any of the methods of claims 1001 to 1032, wherein at the time of administration of said dose of engineered T cells or immediately prior to administration, the subject has relapsed following remission after treatment with three or more prior therapies or has become resistant to said treatment, failed said treatment, and / or was intolerant to said treatment. [The present invention 1034] Any of the methods of claims 1001 to 1033, wherein the one or more prior therapies are selected from a kinase inhibitor, optionally an inhibitor of Bruton's tyrosine kinase (BTK), optionally ibrutinib; venetoclax; combination therapy including fludarabine and rituximab; radiation therapy; and hematopoietic stem cell transplantation (HSCT). [The present invention 1035] The method of any of claims 1001-1035, wherein the one or more prior therapies include an inhibitor of Bruton's tyrosine kinase (BTK) and / or venetoclax. [The present invention 1036] Any of the methods of claims 1001 to 1035, wherein the one or more prior therapies include ibrutinib and / or venetoclax. [The present invention 1037] Any of the methods of claims 1001-1036, wherein the one or more prior therapies include ibrutinib and venetoclax. [The present invention 1038] Any of the methods of claims 1001 to 1037, wherein the subject has relapsed following remission after treatment with, has become resistant to, and / or is intolerant to, an inhibitor of Bruton's tyrosine kinase (BTK) and / or venetoclax. [The present invention 1039] Any of the methods of claims 1001 to 1038, wherein the subject has relapsed following remission after treatment with ibrutinib and venetoclax, has become resistant to them, has failed treatment with them, and / or is intolerant to them. [The present invention 1040] At or prior to administration of said dose of cells, The target is, Optionally, one or more cytogenetic abnormalities associated with high-risk CLL or SLL selected from among complex karyotype or cytogenetic abnormalities, del 17p, unmutated IGVH gene, and TP53 mutations identified or identified as having; and / or The subject is identified or has been identified as having high-risk CLL or SLL, Any of the methods of 1001 to 1039 of the present invention. [The present invention 1041] At or prior to administration of said dose of cells, The target is, Optionally, one or more cytogenetic abnormalities associated with high-risk CLL selected from among complex karyotype or cytogenetic abnormalities, del 17p, unmutated IGVH gene, and TP53 mutation identified or identified as having; and / or The subject is identified or has been identified as having high-risk CLL. Any of the methods according to claims 1001 to 1040. [The present invention 1042] The method of claim 1041, wherein at the time of administration of said dose of cells or immediately prior to administration, the subject has relapsed following remission after treatment with two or more previous therapies or has become resistant to said treatment, has failed said treatment, and / or has been intolerant to said treatment. [The present invention 1043] Any of the methods of claims 1001 to 1040, wherein the subject is identified or has been identified as having standard risk CLL or SLL at the time of or prior to administration of said dose of cells. [The present invention 1044] The method of claim 1043, wherein at the time of administration of said dose of cells or immediately prior to administration, the subject has relapsed following remission after treatment with three or more prior therapies or has become resistant to said treatment, failed said treatment, and / or been intolerant to said treatment. [The present invention 1045] Any of the methods of claims 1001 to 1044, wherein at the time of or prior to administration of said dose of cells, the subject is identified or has been identified as being intolerant to an inhibitor of Bruton's tyrosine kinase (BTK) and has received an inhibitor of BTK for a period of less than or less than about 6 months, and / or is ineligible for treatment with an inhibitor of BTK. [The present invention 1046] (i) the subject is identified or has been identified as having high-risk CLL or SLL, and at the time of or immediately prior to administration of said dose of cells, the subject has relapsed following remission following treatment with one or more prior therapies other than an inhibitor of BTK, or has become resistant to, failed, and / or been intolerant to said treatment; or (ii) the subject is or has been identified as having standard risk CLL or SLL, and at the time of or immediately prior to administration of said dose of cells, the subject has relapsed following remission following treatment with two or more prior therapies other than an inhibitor of BTK, or has become resistant to, failed, and / or been intolerant to said treatment. The method of the present invention 1045. [The present invention 1047] The subject is identified or has been identified as having an ECOG status of 0 or 1; and / or Subjects do not have an ECOG status of >1, Any of the methods of 1001 to 1046 of the present invention. [The present invention 1048] Any of the methods of claims 1001 to 1047, wherein the subject does not have Richter's transformation of CLL or SLL at or immediately prior to administration of said dose of engineered cells or at or immediately prior to administration of lymphodepletion therapy. [The present invention 1049] Any of the methods of claims 1001-1048, wherein the subject is an adult and / or is greater than or about greater than 50, 60, or 70 years of age. [The present invention 1050] The method of any of claims 1001 to 1049, wherein the engineered T cells are primary T cells obtained from the subject. [The present invention 1051] The method of any of claims 1001 to 1050, wherein the engineered T cells are autologous to the subject. [The present invention 1052] The method of any one of claims 1001 to 1051, wherein said dose of engineered cells are viable cells. [The present invention 1053] The method of any of claims 1001 to 1052, further comprising administering to the subject a lymphodepletion therapy prior to administration of said dose of engineered T cells. [The present invention 1054] The method of any one of claims 1001 to 1053, further comprising preconditioning the subject with lymphodepletion therapy. [The present invention 1055] The method of claim 1053 or 1054, wherein the lymphodepleting therapy comprises administration of fludarabine and / or cyclophosphamide. [The present invention 1056] Lymphocyte depletion therapy is performed at approximately 200-400 mg / m 2 (inclusive), optionally 300 or about 300 mg / m 2 of cyclophosphamide, and / or approximately 20-40 mg / m 2 , optionally 30 mg / m 2 6. The method of any of claims 1053 to 1055, comprising daily administration of fludarabine for 2 to 4 days, optionally 3 days. [The present invention 1057] Lymphocyte depletion therapy is at or near 300 mg / m 2 of cyclophosphamide and approximately 30 mg / m 2 of fludarabine daily for three days, optionally wherein said dose of cells is administered at least 2-7 days or at least about 2-7 days after lymphodepletion therapy or at least 2-7 days or at least about 2-7 days after the initiation of lymphodepletion therapy. [The present invention 1058] The method of any of claims 1001 to 1057, wherein administering said dose of engineered T cells and / or administering lymphodepletion therapy is performed via outpatient delivery. [The present invention 1059] The method of any of claims 1001 to 1058, wherein said dose of engineered T cells is administered parenterally, optionally intravenously. [The present invention 1060] Any of the methods of claims 1001 to 1059, wherein the objective response rate is an objective response in at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the subjects treated by the method. [The present invention 1061] Any of the methods of claims 1001 to 1060, wherein the response in at least 35%, at least 40%, at least 50%, at least 60%, or at least 70% of the subjects treated by the method is a complete response (CR). [The present invention 1062] The method of claim 1060 or 1061, wherein the duration of response until progression lasts for more than 3 months or more than 6 months. [The present invention 1063] Any of the methods of claims 1001-1062, wherein more than 50%, more than 60%, or more than 70% of subjects treated by said method had undetectable minimal residual disease (MRD) for at least 1 month, at least 2 months, at least 3 months, or at least 6 months after administration of said dose of engineered T cells. [The present invention 1064] The method of any of claims 1001 to 1063, wherein among a plurality of subjects treated by said method, 10% or less of the subjects exhibit cytokine release syndrome (CRS) greater than grade 2. [The present invention 1065] Any of the methods of claims 1001 to 1064, wherein among a plurality of subjects treated by said method, 10% or less, 20% or less, 30% or less, or 40% or less of the subjects exhibit neurotoxicity greater than grade 2. [The present invention 1066] A plurality of subjects treated by the method include Multiple subjects who have relapsed following remission after treatment with ibrutinib and venetoclax, have become resistant to treatment with them, have failed treatment with them, and / or are intolerant to them Any of the methods of the present invention 1060 to 1064, comprising: [The present invention 1067] assaying a biological sample for a level, amount, or concentration of TNF-α, wherein the biological sample is optionally derived from a subject who is a candidate for treatment with a cell therapy, and the cell therapy comprises a dose of engineered cells, including T cells expressing a CAR for treating a disease or condition, and wherein the biological sample was obtained from the subject prior to administering the cell therapy and / or the biological sample does not comprise a CAR and / or the engineered cells; and comparing the level, amount, or concentration of TNF-α to a threshold level, (1) identifying the subject as at risk for developing grade 3 or greater neurotoxicity following administration of the cell therapy if the level, amount, or concentration of TNF-α is at or above the threshold level; and (2) identifying the subject as not at risk for developing grade 3 or greater neurotoxicity following administration of the cell therapy if the level, amount, or concentration of TNF-α is below the threshold level. A method of treatment comprising: [The present invention 1068] If a subject is identified as being at risk for developing grade 3 or greater neurotoxicity: (i) administering to a subject a cell therapy, optionally at a reduced dose; and optionally (a) the method further comprises administering to the subject an agent or other treatment capable of treating, preventing, delaying, reducing or attenuating the onset or risk of onset of neurotoxicity; and / or (b) administration of the cellular therapy to the subject is performed or designated to be performed in an inpatient setting and / or with a one or more day hospital stay; stage; or (ii) administering to the subject an alternative treatment other than cell therapy to treat the disease or condition. The method of claim 1067, further comprising: [The present invention 1069] If a subject is identified as not at risk for developing grade 3 or greater neurotoxicity following administration of the cell therapy, (i) optionally, at or after the subject exhibits persistent fever or a fever that has or has not decreased by more than 1° C. following treatment with an antipyretic, unless or until the subject exhibits signs or symptoms of toxicity, the subject is not administered an agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset of, or the risk of the onset of, toxicity; and / or (ii) optionally, the administration and all follow-up is performed on an outpatient basis and / or without admitting the subject to a hospital and / or without an overnight stay in a hospital and / or without the need for hospitalization or overnight stay in a hospital, unless or until the subject exhibits persistent fever or a fever that has or has not decreased by more than 1° C. following treatment with antipyretics; The method of the present invention 1067. [The present invention 1070] The assaying step comprises: (a) contacting the biological sample with one or more reagents capable of detecting TNF-α or specific for TNF-α, optionally wherein the one or more reagents comprise an antibody that specifically recognizes TNF-α; and (b) detecting the presence or absence of a complex comprising one or more reagents and TNF-α. Any of the methods of claims 1067 to 1069, comprising: [The present invention 1071] A method of treatment comprising administering to a subject a cell therapy for treating a disease or condition, the cell therapy comprising a dose of engineered cells comprising T cells expressing a CAR; (1) a subject is identified as being at risk for developing grade 3 or greater neurotoxicity following administration of a cell therapy if the subject has a level, amount, or concentration of TNF-α in a biological sample from the subject that is at or above a threshold level; (i) administering the cell therapy at a reduced dose to the subject; (ii) further administering to the subject an agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset or risk of the onset of toxicity; and / or (iii) administering to the subject the cell therapy is or is designated to be administered in an inpatient setting and / or with a one or more day hospitalization; or (2) if a subject is selected or identified as having a level, amount, or concentration of TNF-α in a biological sample from the subject that is below a threshold level, the subject is identified as not at risk for developing Grade 3 or greater neurotoxicity following administration of the cell therapy, wherein: (i) optionally, at or after the subject exhibits persistent fever or fever that falls or does not fall or does not fall by more than 1° C. following treatment with antipyretics, the subject is not administered any agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset of, or the risk of the onset of, toxicity, unless or until the subject exhibits signs or symptoms of toxicity; and / or (ii) optionally, unless or until the subject exhibits persistent fever or fever that falls or does not fall or does not fall by more than 1° C. following treatment with antipyretics, the administration and all follow-up is performed on an outpatient basis and / or without the subject being admitted to a hospital and / or without an overnight stay in a hospital and / or without the need for hospitalization or overnight stay in a hospital, the subject is a candidate for treatment with a cell therapy and the biological sample obtained from the subject prior to administration of the cell therapy and / or the biological sample does not contain a CAR and / or the engineered cells; The method. [The present invention 1072] Any of the methods of inventions 1068, 1070 and 1071, wherein if the subject is identified as at risk for developing grade 3 or greater neurotoxicity following administration of the cell therapy, an agent or other treatment capable of treating, preventing, delaying, reducing or attenuating the onset or risk of onset of toxicity is administered, wherein the agent is administered to the subject simultaneously with the cell therapy or within 3 days of administration of the cell therapy to the subject. [The present invention 1073] the threshold level is within 25%, 20%, 15%, 10%, or 5% above the median or mean value of a level, amount, or concentration of TNF-α in a biological sample obtained from a group of subjects prior to receiving cell therapy, and / or is within a standard deviation above said median or mean value, or is at or about the median or mean value of said level, amount, or concentration, and each of the subjects in the group did not exhibit any grade of neurotoxicity following administration of a dose of engineered cells expressing a CAR to treat the same disease or condition; the threshold level is 1.25-fold or greater than 1.25-fold greater than the median or mean level, amount, or concentration of TNF-α in biological samples obtained from a group of subjects prior to receiving cell therapy, and each of the subjects in the group did not exhibit any grade of neurotoxicity following administration of a dose of engineered cells expressing a CAR to treat the same disease or condition; the threshold level is 1.25-fold or more than 1.25-fold higher than the level, amount, or concentration of TNF-α in a biological sample obtained from a group of normal or healthy subjects who are not candidates for cell therapy treatment; Any of the methods of 1067 to 1072 of the present invention. [The present invention 1074] (a) assaying a biological sample from a subject receiving a cell therapy comprising a dose of engineered cells comprising T cells expressing a CAR to treat a disease or condition for the level, amount, or concentration of IL-16, wherein the biological sample is obtained from the subject within 1, 2, or 3 days after initiation of administration of the cell therapy; and (b) comparing the level, amount, or concentration of IL-16 to a threshold level, (1) identifying the subject as at risk for developing grade 3 or higher neurotoxicity if the level, amount, or concentration of IL-16 is at or above the threshold level; and (2) identifying the subject as not at risk for developing grade 3 or higher neurotoxicity if the level, amount, or concentration of IL-16 is below the threshold level. A method of treatment comprising: [The present invention 1075] The method of the present invention 1074, wherein if the subject is identified as being at risk for developing grade 3 or higher neurotoxicity, an agent or other treatment capable of treating, preventing, delaying, reducing, or attenuating the onset or risk of onset of toxicity is administered. [The present invention 1076] The assaying step comprises: (a) contacting the biological sample with one or more reagents capable of detecting IL-16 or specific for IL-16, optionally wherein the one or more reagents comprise an antibody that specifically recognizes IL-16; and (b) detecting the presence or absence of a complex comprising one or more reagents and IL-16. The method of the present invention 1074 or 1075, [The present invention 1077] The method of any one of claims 1074 to 1076, further comprising administering a cell therapy to the subject prior to the assaying step. [The present invention 1078] administering to a subject identified as being at risk for developing grade 3 or greater neurotoxicity an agent or other treatment capable of treating, preventing, delaying, reducing, or attenuating the onset or risk of the onset of toxicity; the subject has previously received a cell therapy to treat a disease or condition; the subject is selected or identified as being at risk for developing grade 3 or greater neurotoxicity at or shortly before administration of the agent if the level or amount or concentration of IL-16 in a biological sample obtained from the subject within 1, 2, or 3 days of initiating administration of the cell therapy is above a threshold level; step A method of treatment comprising: [The present invention 1079] Any of the methods of claims 1075 to 1078, wherein the step of administering the agent is performed when the subject exhibits persistent fever or a fever that has decreased by more than 1°C or has not decreased or does not decrease after treatment with an antipyretic agent. [The present invention 1080] Any of the methods of claims 1075 to 1079, wherein administration of the cell therapy to the subject is performed on an outpatient basis and, if the level, amount, or concentration of IL-16 exceeds a threshold level, the method includes a step of hospitalizing the patient in a hospital for one or more days. [The present invention 1081] Any of the methods of inventions 1074-1080, wherein the threshold level is within 25% above the median or mean value of the level, amount, or concentration of IL-16 in biological samples obtained from a group of subjects within 1, 2, or 3 days after receiving a cell therapy comprising administration of a dose of engineered cells expressing a CAR to treat the same disease or condition, and / or is within a standard deviation above said median or mean value, or is at or about said median or mean value of said level, amount, or concentration, and each of the subjects in the group did not exhibit any grade of neurotoxicity after administration of the cell therapy. [The present invention 1082] Any of the methods of claims 1074-1080, wherein the threshold level is 1.3-fold or greater than 1.3-fold greater than the median or mean level, amount, or concentration of IL-16 in biological samples obtained from a group of subjects prior to receiving cell therapy, and each of the subjects in the group did not exhibit any grade of neurotoxicity following administration of a dose of engineered cells expressing a CAR to treat the same disease or condition. [The present invention 1083] Any of the methods of claims 1074 to 1080, wherein the threshold level is 1.3-fold or more than 1.3-fold higher than the level, amount, or concentration of IL-16 in a biological sample obtained from a group of normal or healthy subjects who are not candidates for cell therapy treatment. [The present invention 1084] The method of any of claims 1067 to 1083, wherein the biological sample is or is obtained from a blood, plasma, or serum sample. [The present invention 1085] The method of any one of claims 1067 to 1084, wherein the assaying step comprises an immunoassay. [The present invention 1086] The method of any one of claims 1067 to 1085, wherein the disease or condition is cancer. [The present invention 1087] The method of any one of claims 1067 to 1086, wherein the disease or condition is myeloma, leukemia, or lymphoma. [The present invention 1088] Biological samples were analyzed for ROR1, B-cell maturation antigen (BCMA), carbonic anhydrase 9 (CAIX), tEGFR, Her2 / neu (receptor tyrosine kinase erbB2), L1-CAM, CD19, CD20, CD22, mesothelin, CEA, and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimer, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-α, IL-13R-α2, kinase insert domain receptor (kdr), kappa light chain, Lewis Y, L1 cell adhesion molecule, (L1-CAM), melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, preferentially expressed antigen in melanoma (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250 / CAIX, HLA-AI MAGE Al, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7 / 8, avb6 integrin, 8H9, NCAM, VEGF receptor, 5T4, fetal AchR, NKG2D ligand, CD44v6, bispecific antigen, cancer testis antigen, mesothelin, mouse CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, oncofetal antigen, ROR1, TAG72, VEGF-R2, Any of the methods of claims 1067 to 1087, comprising an antigen that is carcinoembryonic antigen (CEA), Her2 / neu, estrogen receptor, progesterone receptor, ephrin B2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms tumor 1 (WT-1), cyclin, cyclin A2, CCL-1, CD138, G protein-coupled receptor 5D (GPCR5D), or a pathogen-specific antigen. [The present invention 1089] The method of any of claims 1067 to 1088, wherein the biological sample comprises an antigen which is CD19. [The present invention 1090] Any of the methods of claims 1067 to 1089, wherein the disease or condition is a B-cell malignancy and / or is acute lymphoblastic leukemia (ALL), adult ALL, chronic lymphoblastic leukemia (CLL), small lymphocytic lymphoma (SLL), non-Hodgkin's lymphoma (NHL), and diffuse large B-cell lymphoma (DLBCL). [The present invention 1091] The method of any one of claims 1067 to 1090, wherein the disease or condition is CLL or SLL. [The present invention 1092] The method of any of claims 1068 to 1073 and 1075 to 1091, wherein the agent or other treatment is or comprises an anti-IL-6 antibody, an anti-IL-6R antibody, or a steroid. [The present invention 1093] The method of any of claims 1068 to 1073 and 1075 to 1092, wherein the agent is or comprises tocilizumab, siltuximab, or dexamethasone. [The present invention 1094] CAR, an extracellular antigen-binding domain specific for CD19; A transmembrane domain; a cytoplasmic signaling domain derived from a costimulatory molecule, optionally 4-1BB; a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule, optionally CD3ζ; Includes; the CAR, in turn, comprises an extracellular antigen-binding domain specific for CD19, a transmembrane domain, a cytoplasmic signaling domain derived from a costimulatory molecule, and a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule; Any of the methods of 1001 to 1093 of the present invention. [The present invention 1095] The method of the present invention, wherein the antigen-binding domain is an scFv. [The present invention 1096] the scFv comprises a CDRL1 sequence of RASQDISKYLN (SEQ ID NO: 35), a CDRL2 sequence of SRLHSGV (SEQ ID NO: 36), and / or a CDRL3 sequence of GNTLPYTFG (SEQ ID NO: 37), and / or a CDRH1 sequence of DYGVS (SEQ ID NO: 38), a CDRH2 sequence of VIWGSETTYYNSALKS (SEQ ID NO: 39), and / or a CDRH3 sequence of YAMDYWG (SEQ ID NO: 40); the scFv comprises the variable heavy chain region of FMC63 and the variable light chain region of FMC63, and / or the CDRL1 sequence of FMC63, the CDRL2 sequence of FMC63, the CDRL3 sequence of FMC63, the CDRH1 sequence of FMC63, the CDRH2 sequence of FMC63, and the CDRH3 sequence of FMC63, or binds to the same epitope as any of the foregoing or competes for binding with any of the foregoing; the scFv comprises a VH as set forth in SEQ ID NO:41 and a VL as set forth in SEQ ID NO:42, optionally separated by a flexible linker, optionally the flexible linker being or comprising the sequence as set forth in SEQ ID NO:24; and / or The scFv is or comprises the sequence shown in SEQ ID NO:43. The method of the present invention 1095. [The present invention 1097] The method of any of claims 1094 to 1096, wherein the costimulatory signaling region is a signaling domain of CD28 or 4-1BB. [The present invention 1098] The method of any one of claims 1094 to 1097, wherein the costimulatory signaling region is a signaling domain of 4-1BB. [This invention 1099] Any of the methods of claims 1094 to 1098, wherein the costimulatory domain comprises SEQ ID NO: 12, or a variant thereof having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto. [The present invention 1100] The method of any of claims 1094 to 1099, wherein the primary signaling domain is the CD3 zeta signaling domain. [The present invention 1101] The method of any of claims 1094-1100, wherein the primary signaling domain comprises SEQ ID NO: 13 or 14 or 15 having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto. [The present invention 1102] The method of any of claims 1094 to 1101, wherein the CAR further comprises a spacer between the transmembrane domain and the scFv. [The present invention 1103] 1103. The method of claim 1102, wherein the spacer is a polypeptide spacer comprising or consisting of all or part of an immunoglobulin hinge or a modified version thereof, optionally an IgG4 hinge or a modified version thereof. [The present invention 1104] The method of claim 1102 or 1103, wherein the spacer is about 15 amino acids or less and does not include the CD28 extracellular region and the CD8 extracellular region. [The present invention 1105] The method of any of claims 1102 to 1104, wherein the spacer is 12 amino acids or about 12 amino acids in length. [The present invention 1106] The spacer is having or consisting of a sequence encoded by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, or a variant of any of the foregoing having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto; and / or X 1 is glycine, cysteine, or arginine and X 2 is cysteine ​​or threonine; 1 PPX 2 comprising or consisting of P, Any of the methods 1102 to 1105 of the present invention. [The present invention 1107] The method of any one of claims 1001 to 1106, wherein the subject is a human subject. [The present invention 1108] An article of manufacture comprising a cell therapy composition or one of a plurality of cell therapy compositions comprising T cells expressing an anti-CD19 chimeric antigen receptor (CAR) and instructions for administering the cell therapy, the instructions specifying a step of administering the T cell composition according to any of the methods of the present invention 1001-1106. [Brief description of the drawings]

[0068] [Figure 1] Figure 1 shows a box plot of CAR T cell frequencies in whole blood measured by flow cytometry. Boxes span the first through third quartiles, with the median shown as a horizontal line. Whiskers span 1.5 times the interquartile range, with measurements outside this range plotted as individual points. [Diagram 2] A graph of median cells / μl over time by dose level is shown. Patients, N=16; Samples, n. Upper error bars represent 3rd quartile; lower error bars represent 1st quartile. Doses were given on day 1. [Diagram 3] Figures 3A and 3B show graphs of biomarker analysis of IL-16 and TNFα over time in patients. Patients, N=13. 39 cytokines were analyzed in 13 patients within the first 30 days of treatment. [Figure 4]Figure 4A shows best overall response results in subjects (N=22) with R / R CLL administered anti-CD19 CAR+T cells at DL1 (5×107 CAR-expressing T cells) or DL2 (1×108 CAR-expressing T cells). Figure 4B shows minimal residual disease undetectable (uMRD) results in blood by flow cytometry or uMRD results in bone marrow by next generation sequencing (NGS) at any time point following administration of anti-CD19 CAR-expressing T cells at DL1 (5×107 CAR-expressing T cells) or DL2 (1×108 CAR-expressing T cells) to subjects with R / R CLL. [Diagram 5] Shown is a swimmer plot of duration of response over time in subjects (N=22) with R / R CLL administered anti-CD19 CAR+ T cells in DL1 (5×107 CAR-expressing T cells) or DL2 (1×108 CAR-expressing T cells). [Figure 6] Figure 1 shows a graph of median cells / μl over time by dose level in subjects with R / R CLL who received anti-CD19 CAR+T cells at DL1 (5×107 CAR-expressing T cells) or DL2 (1×108 CAR-expressing T cells). The upper error bar represents the third quartile and the lower error bar represents the first quartile. The dose of anti-CD19 CAR+T cells was given on day 1. [Figure 7]Figure 7A shows the results of the best overall response in subjects with R / R CLL (N = 22) or subjects (N = 9) who had failed previous treatment with both a Bruton's tyrosine kinase inhibitor (BTKi) and venetoclax. Figure 7B shows the results of undetectable minimal residual disease (uMRD) in blood by flow cytometry or uMRD in bone marrow by next-generation sequencing (NGS) at any time point after dosing in subjects with R / R CLL (N = 20) or subjects (N = 8) who had failed previous treatment with both BTKi and venetoclax. a Evaluable for response was defined as receiving a pre-treatment assessment and ≥1 post-baseline assessment; evaluable for MRD was defined as patients with detectable MRD at baseline. One subject was not evaluable for response. b Failure of venetoclax was defined as PD after ≥3 months of treatment or discontinuation due to <PR. c Two subjects were not evaluable for MRD. d One subject was not evaluable for MRD. CI, confidence interval; CRi, complete response with incomplete blood count recovery; NGS, next-generation sequencing; nPR, nodular partial response; PD, progression; PR, partial response; SD, stable; uMRD, undetectable minimal residual disease. [Figure 8] Swimmer plots of duration of response over time in individual subjects with R / R CLL who had failed previous treatment with both BTKi and venetoclax, and other treated subjects. * MRD evaluation not available. Seven deaths occurred during the trial: five subjects died due to disease progression; one subject had grade 5 respiratory failure (DL1) unrelated to CAR + T cell therapy treatment; one subject had septic shock, acute kidney injury, and pneumonia (DL2) unrelated to CAR + T cell therapy treatment. No deaths occurred within the first 30-day period. ND, not performed; RT, Richter transformation. [Figure 9]Graphs of median cells / μl over time by dose level in evaluable treated subjects and in subjects who failed prior treatment with both BTKi and venetoclax. Upper error bars represent 3rd quartile and lower error bars represent 1st quartile. CAR+T cell therapy was given on day 1. AUC0-29, area under the curve from days 0 to 29; Cmax, maximum concentration; PK / PD, pharmacokinetics / pharmacodynamics; Q, quartiles; Tmax, time to maximum concentration. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0069] Detailed Description Provided herein are methods and uses of engineered cells (e.g., T cells) and / or compositions thereof for the treatment of subjects with a disease or condition that is or includes, generally, cancer or tumor, e.g., leukemia or lymphoma, most particularly chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL). In some aspects, the methods and uses result in or achieve improved response and / or sustained response or efficacy and / or reduced risk of toxicity or other side effects, for example, in a particular group of subjects being treated, compared to certain alternative methods. In some embodiments, the methods are advantageous by administering a specified number or relative number of engineered cells, administering a defined ratio of certain types of cells, treating a particular patient population, for example, a patient population with a particular risk profile, stage classification, and / or previous therapy history, and / or combinations thereof.

[0070] Also provided are methods that include evaluating certain parameters, such as the expression or analytes of specific biomarkers, that can be correlated with the onset of toxicity, and methods for treatment, such as interventional therapy, to prevent and / or ameliorate toxicity.Also provided are methods that involve evaluating certain parameters, such as the expression or analytes of specific biomarkers, that can be correlated with outcomes, such as treatment outcomes, including responses, such as complete response (CR) or partial response (PR); or safety outcomes, such as toxicity, such as the onset of neurotoxicity or CRS, after administration of immunotherapy and / or cell therapy.Also provided are methods for assessing the likelihood of response and / or the likelihood of risk of toxicity based on the evaluation of parameters, such as the expression or analytes of biomarkers.Also provided are compositions for use in cell therapy.Also provided are articles of manufacture and kits, for example, for use in the methods provided herein.In some embodiments, the articles of manufacture and kits optionally include instructions for use in the methods provided herein.

[0071] Specifically, among the embodiments provided are methods for treating subjects with CLL or SLL. In some aspects, CLL is considered an incurable disease, and subjects eventually relapse or become resistant to available treatments or treatments. In some of any of the embodiments, the subject has high-risk disease. In some embodiments of the methods provided, the subject has high-risk CLL or SLL. In some cases, existing treatment strategies for high-risk and very high-risk subjects may include fludarabine, cyclophosphamide, and rituximab (FCR), Bruton's tyrosine kinase (BTK) inhibitors (e.g., ibrutinib), and / or allogeneic stem cell transplantation (Puiggros et al., BioMed Research International, Volume 2014 (2014), Article ID 435983). Many of the existing therapies include oral targeted drugs, which have improved treatment outcomes in some patients with CLL. Nevertheless, some patients are intolerant or resistant to therapy and / or do not achieve complete response with undetectable MRD (uMRD). In some aspects, subjects with progressive disease after treatment with available therapy have poor outcomes. For example, in some aspects, subjects treated for CLL show poor long-term outcomes. For example, in some cases, resistant (R / R) high-risk CLL subjects have poor survival rates after stopping ibrutinib (Jain et al. (2015) Blood 125(13):2062-2067). There is a need for improved methods of treating CLL, in some aspects suitable for treating high-risk and / or very high-risk CLL and / or subjects who have relapsed or become resistant to multiple previous therapies.

[0072] In some embodiments, the method comprises administering cells to a subject selected or identified as having a certain prognosis or risk of CLL. Chronic lymphocytic leukemia (CLL) is generally a variable disease. Some subjects with CLL survive without treatment, while others require immediate intervention. In some cases, subjects with CLL may be classified into groups that may characterize disease prognosis and / or recommended treatment strategies. In some cases, these groups may be "low risk", "intermediate risk", "high risk", and / or "very high risk", and patients may be classified as such depending on multiple factors, including but not limited to genetic abnormalities and / or morphological or physical characteristics. In some embodiments, the subject treated by the method is classified or identified based on the risk of CLL. In some embodiments, the subject is a subject with high-risk CLL.

[0073] In some embodiments, the provided methods and uses provide or achieve improved or more durable response or efficacy compared to certain alternative methods, for example, in certain groups of subjects treated, for example, in patients with leukemia such as CLL or SLL, including subjects with high-risk disease.In some embodiments, the method is advantageous by administering a composition that includes cells for adoptive cell therapy, such as T cell therapy, for example, CAR-expressing T cells, for example, anti-CD19 CAR+ T cells.In some embodiments, the method also includes lymphodepletion therapy, for example, cyclophosphamide, fludarabine, or a combination thereof, before T cell therapy.

[0074] In some aspects, the provided embodiments are based on the observation that the provided methods can be used to achieve high response rates with high durability without increased risk of toxicity, as compared to certain available methods for cell therapy, for example, the observation described in the examples provided herein.In some embodiments, the provided methods allow for extended survival of adoptively transferred cells for cell therapy, and / or low toxicity incidence in subjects.In some embodiments, the methods can be used to select subjects for treatment with cell therapy who are likely or more likely to respond to cell therapy, and / or to determine appropriate doses or administration regimens for higher response rates and / or more durable responses, while minimizing the risk of toxicity.Such methods can provide information for rational strategies to facilitate safe and effective clinical application of adoptive cell therapy, such as CAR-T cell therapy.

[0075] In some embodiments, the provided methods achieve high response rates in a highly treated population of subjects with high-risk CLL (or SLL), all of whom have received one or more previous therapies, including ibrutinib. In some embodiments, the subjects treated include those who have relapsed after first remission with ibrutinib, or those who are resistant or intolerant to treatment with ibrutinib. In certain embodiments, the subjects treated include those who have relapsed after remission, or those who are resistant or intolerant to one or more additional previous therapies, such as 1, 2, 3, 4, 5 or more previous therapies in addition to ibrutinib. In some embodiments, the subjects relapsed or are resistant to both ibrutinib and venetoclax pretreatment. In some embodiments, the subjects who are resistant to such treatment have progressed after one or more previous therapies. In some embodiments, subjects to be treated, including those treated with one or more previous therapies (e.g., ibrutinib and / or venetoclax), include those with high-risk cytogenetic abnormalities, including TP53 mutations, complex karyotypes (i.e., at least three chromosomal alterations), and del17(p). In some embodiments, subjects for treatment according to the embodiments provided herein include those who have failed both BTK inhibitors (e.g., ibrutinib) and venetoclax. As disclosed herein, results from ongoing clinical trials show high overall response rates (ORR) of more than 65% of treated subjects across dose levels, including complete remission (CR) with incomplete blood count recovery (CRi) in more than 35% of treated subjects. Of these subjects, all had been previously treated with ibrutinib, and approximately half had been previously treated with ibrutinib and venetoclax. In some aspects, outcome is associated with achievement of undetectable MRD (uMRD); achievement of uMRD has been reported to correlate with improved outcomes (Kovacs et al. (2016) J. Clin. Oncol., 34:3758-3765; Thompson and Wierda (2016) Blood, 127:279-286).In some embodiments, the methods provided result in a high percentage of durable responses that last for more than 1 month, more than 3 months, more than 6 months, or more, without progression.

[0076] Such results achieved in high-risk subjects are superior compared to certain other alternative therapies.In particular, CLL is generally considered to be incurable, and patients often eventually relapse or become resistant to available treatments (Dighiero and Hamblin (2008) The Lancet, 371:1017-1029).In some cases, CR and uMRD are insufficient, and / or subjects progress or have poor outcomes after treatment with certain other agents, such as ibrutinib as a single agent, venetoclax-rituximab, bendamustine-rituximab, or both ibrutinib and venetoclax.Moreover, multiple reports suggest that certain other CAR T cell therapies cannot achieve such sustained response rates.

[0077] In some embodiments, the method and use comprise administering to a subject cells expressing engineered (recombinant) cell surface receptors in adoptive cell therapy, the receptors are generally chimeric receptors, such as chimeric antigen receptors (CARs), that recognize antigens expressed by, associated with, and / or specific to leukemia or lymphoma and / or cell types derived therefrom.In certain embodiments, the antigen targeted is CLL.The cells are generally administered in a composition formulated for administration; the method generally involves administering one or more doses of cells to a subject, and the doses may comprise a specific or relative number of cells or engineered cells, and / or a defined ratio or composition of two or more subtypes, such as CD4 T cells:CD8 T cells, in the composition.

[0078] In certain embodiments, the method is carried out by a therapeutic T cell product with separate administration of CD4+ and CD8+ CAR T cell compositions, where CD4+ and CD8+ CAR T cells are administered in specific or exact numbers as a flat dose and / or as a defined ratio. In some cases, the method includes creating or manipulating the CAR T cell composition by a process that includes separate isolation, selection, or enrichment of CD4+ and CD8+ T cells from a biological sample. In some cases, the method for creating a CAR-T cell composition that includes enrichment of CD4+ and CD8+ T cells avoids the risk of including tumor cells in the CAR-T cell product or during the manufacture of the CAR-T cell product. Compared to other diseases, CLL is a cancer in which tumor cells are found in the periphery, which in some circumstances may prevent and / or affect the efficacy of a CAR-T product that may include such cells or may be derived from an initial composition containing such cells.

[0079] All publications, including patent documents, scientific literature, and databases, referred to in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication was individually incorporated by reference. To the extent that the definitions set forth herein differ or are otherwise inconsistent with those set forth in the patents, applications, published applications, and other publications incorporated herein by reference, the definitions set forth herein take precedence over the definitions incorporated herein by reference.

[0080] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described.

[0081] I. Methods and Uses of Cellular Therapy with Genetically Engineered Cells In some embodiments, the methods and uses provided herein include administering to a subject cells expressing engineered (recombinant) cell surface receptors in adoptive cell therapy, which generally are chimeric receptors, such as chimeric antigen receptors (CARs), that recognize antigens expressed by, associated with, and / or specific to leukemia or lymphoma and / or cell types derived therefrom.The cells are generally administered in a composition formulated for administration; the method generally involves administering one or more doses of cells to a subject, which may include a specific or relative number of cells or engineered cells, and / or a defined ratio or composition of two or more subtypes, such as CD4 T cells:CD8 T cells, in the composition.

[0082] In some embodiments, the cells, populations, and compositions are administered to a subject having a particular disease or condition to be treated, for example, via adoptive cell therapy, such as adoptive T cell therapy. In some embodiments, the methods involve treating a subject having a lymphoma or leukemia, such as chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL), with a dose of antigen receptor-expressing cells (e.g., CAR-expressing cells).

[0083] In some embodiments, the methods provided involve treating a particular group or subset of subjects, such as subjects identified as having high-risk disease, such as high-risk CLL. In some aspects, the methods treat subjects with a form of aggressive and / or poor prognosis CLL, such as CLL that has relapsed or is resistant to standard treatment (R / R) and has a poor prognosis. In some embodiments, the subject has failed one or more previous therapies. In some embodiments, the subject is ineligible for other previous therapies. In some embodiments, the subject has failed previous therapy with a Bruton's tyrosine kinase inhibitor (BTKi), such as ibrutinib. In some embodiments, the subject has failed ibrutinib and venetoclax. In some cases, the overall response rate (ORR; also known in some cases as objective response rate) is less than 40% and / or the complete response (CR; also known in some cases as complete remission) is less than 20% relative to available treatments, to a standard treatment, or to a reference treatment for the disease and / or patient population for which the treatment is indicated.

[0084] In some embodiments, the methods, uses, and articles of manufacture involve or are used for treating subjects with, selecting, or identifying a particular group or subset of subjects based on, for example, a particular disease type, diagnostic criteria, previous treatment, and / or response to previous treatment. In some embodiments, the methods involve treating subjects who have relapsed or become resistant to treatment following remission after treatment with one or more previous therapies; or subjects who have relapsed or become resistant to one or more previous therapies, e.g., one or more lines of standard treatment (R / R). In some embodiments, the methods involve treating subjects with chronic lymphocytic leukemia. In some embodiments, the methods involve treating subjects with small lymphocytic lymphoma. In some embodiments, the methods involve treating subjects with an Eastern Cooperative Oncology Group Performance Status (ECOG) of 0-1. In some embodiments, the methods treat poor prognosis populations of CLL patients or subjects who have an inadequate response to treatments in general or to specific reference treatments, such as populations with high-risk cytogenetic abnormalities (i.e., Del(17p), TP53 mutations, mutant IGHV, and complex karyotypes).

[0085] In some embodiments, the antigen receptor (e.g., CAR) specifically binds to a target antigen associated with a disease or condition, for example, associated with CLL. In some embodiments, the antigen receptor binds to a target antigen associated with SLL. In some embodiments, the antigen associated with a disease or disorder is CD19.

[0086] In some embodiments, the method comprises administering a cell or a composition containing a cell to a subject, tissue, or cell, for example, having, at risk of, or suspected of having a disease, condition, or disorder.In some embodiments, the subject is an adult subject.In some embodiments, the subject is more than 50, 60, or 70 years old, or about 50, 60, or 70 years old.

[0087] In some embodiments, the subject has been previously treated with a therapy or therapeutic agent targeting a disease or condition, such as CLL or SLL, prior to administration of the cells expressing the recombinant receptor. In some embodiments, the subject has been previously treated with hematopoietic stem cell transplantation (HSCT), such as allogeneic HSCT or autologous HSCT. In some embodiments, the subject has had a poor prognosis after treatment with standard therapy and / or has failed one or more lines of previous therapy, such as at least 1, 2, 3, 4 or more lines, or at least about 1, 2, 3, 4 or more lines of previous therapy. In some embodiments, the subject is treated with or has previously received at least 1, 2, 3, or 4, or at least about 1, 2, 3, or 4, or about 1, 2, 3, or 4 other therapies for treating CLL, other than lymphodepleting therapy and / or a dose of cells expressing the antigen receptor. In some embodiments, the subject has been previously treated with chemotherapy or radiation therapy. In some aspects, the subject is resistant or non-responsive to other therapies or therapeutic agents.In some embodiments, the subject has persistent or recurrent disease after treatment with another therapy or therapeutic intervention, including, for example, chemotherapy or radiation.In some embodiments, the subject has relapsed or resistant (R / R) chronic lymphocytic leukemia (CLL) and has failed or is ineligible for Bruton's tyrosine kinase inhibitor (BTKi) treatment.

[0088] In some embodiments, the subject has been previously treated with a therapy or therapeutic agent targeted to a disease or condition, e.g., CLL, prior to administration of the cells expressing a recombinant antigen receptor. In some embodiments, the therapeutic agent is a kinase inhibitor, such as an inhibitor of Bruton's tyrosine kinase (Btk), e.g., ibrutinib. In some embodiments, the therapeutic agent is an inhibitor of B-cell lymphoma-2 (Bcl-2), e.g., venetoclax. In some embodiments, the therapeutic agent is an antibody (e.g., a monoclonal antibody) that specifically binds to an antigen expressed by cells of CLL or NHL, e.g., an antigen from any one or more of CD20, CD19, CD22, ROR1, CD45, CD21, CD5, CD33, Igκ, Igλ, CD79a, CD79b, or CD30. In some embodiments, the therapeutic agent is an anti-CD20 antibody, e.g., rituximab. In some embodiments, the therapeutic agent is a depleting chemotherapy that is a combination therapy that includes rituximab, for example, a combination therapy of fludarabine and rituximab or a combination therapy of anthracycline and rituximab.In some embodiments, the subject has been previously treated with hematopoietic stem cell transplantation (HSCT), for example, allogeneic HSCT or autologous HSCT.In some embodiments, the subject has been previously treated with or has previously received at least 1, 2, 3, or 4, or at least about 1, 2, 3, or 4, or about 1, 2, 3, or 4 other therapies for treating CLL, other than lymphodepleting therapy and / or a dose of cells expressing antigen receptor.In some embodiments, the subject has been previously treated with chemotherapy or radiation therapy.

[0089] In some aspects, the subject is resistant or non-responsive to other therapies or therapeutic agents. In some embodiments, the subject has persistent or recurrent disease after treatment with another therapy or intervention, including, for example, chemotherapy or radiation.

[0090] In some embodiments, the subject is a subject that is eligible for transplantation, e.g., a subject that is eligible for hematopoietic stem cell transplantation (HSCT), e.g., allogeneic HSCT. In some such embodiments, the subject is eligible but has not previously undergone a transplantation prior to administration of the engineered cells (e.g., CAR-T cells) or compositions comprising the cells to the subject as provided herein.

[0091] In some embodiments, the subject is a subject who is not eligible for transplantation, e.g., a subject who is not eligible for hematopoietic stem cell transplantation (HSCT), e.g., allogeneic HSCT. In some embodiments, such a subject is administered an engineered cell (e.g., a CAR-T cell) or a composition comprising the cell according to the embodiments provided herein.

[0092] In some embodiments, the method includes administering cells to a subject selected or identified as having high-risk CLL. In some embodiments, the subject exhibits one or more cytogenetic abnormalities, e.g., associated with high-risk CLL. In some aspects, the treated population includes subjects with an Eastern Cooperative Oncology Group Performance Status (ECOG) of 0-1.

[0093] In some aspects of any of the embodiments, the subject to be treated has failed two or more previous therapies. In some aspects of any of the embodiments, the subject to be treated has failed three or more previous therapies. In some embodiments, the previous therapies include any of the following: an inhibitor of Bruton's tyrosine kinase (BTK), such as ibrutinib; venetoclax; combination therapy including fludarabine and rituximab; radiation therapy; and hematopoietic stem cell transplant (HSCT). In some embodiments, the subject or patient has previously been treated with ibrutinib and / or venetoclax, but has relapsed after remission, is resistant to the treatment, has failed the treatment, and / or is intolerant to the treatment. In some embodiments, the subject or patient has previously been treated with ibrutinib and venetoclax, but has relapsed after remission, is resistant to the treatment, has failed the treatment, and / or is intolerant to the treatment.

[0094] In some embodiments, methods are provided for treating subjects selected or identified as relapsing after remission or resistant to pretreatment with ibrutinib and venetoclax for treating CLL or SLL. In some aspects, the selected or identified subjects are administered CAR T cell therapy, such as anti-CD19 CAR-T cell therapy, according to the methods provided.

[0095] In some embodiments, the subject has never achieved a complete response (CR), has not undergone an autologous stem cell transplant (ASCT), is refractory to one or more second line therapies, has primary resistant disease, and / or has an ECOG performance score of 0-1.

[0096] In some aspects, subjects treated according to the provided embodiments include those with a diagnosis of CLL or SLL. In some embodiments, subjects with CLL are treated according to the International Workshop on Chronic Lymphocytic Leukemia (iwCLL) guidelines and clinically measurable disease (bone marrow infiltration with > 30% lymphocytes, peripheral blood lymphocytosis > 5 x 10 9 In some embodiments, subjects with CLL include those with a diagnosis of CLL with indications for treatment based on measurable disease (defined as at least one lesion > 1.5 cm in greatest diameter, lymphadenopathy and / or splenomegaly, and / or measurable lymph nodes and / or liver or splenomegaly) at the time of diagnosis, and < 5 x 10 in peripheral blood. 9 Included are subjects with a diagnosis of SLL based on CD19+ CD5+ clonal B lymphocytes / L [< 5000 / μL] and subjects with biopsy-diagnosed SLL.

[0097] In some aspects, the subject is ineligible for treatment with a Bruton's tyrosine kinase inhibitor (BTKi, e.g., ibrutinib) due to the requirement of full-dose anticoagulants or treatment history or arrhythmia, or has previously failed treatment after receiving a BTKi as determined by stable disease (SD) or progressive disease (PD) as best response, PD after a previous response, or discontinuation due to intolerance (e.g., unmanageable toxicity). In some aspects, the subject is treated according to the provided embodiments if the subject has high-risk disease (determined by complex cytogenetic abnormalities (e.g., complex karyotype), del(17p), TP53 mutation, unmutated IGVH) and has failed (e.g., at least) two or more previous therapies; or if the subject has standard-risk disease and has failed (e.g., at least) three or more previous therapies. In some aspects, subjects treated according to the provided embodiments exclude subjects with untreated active CNS disease, ECOG>1, or Richter transformation.

[0098] In some aspects, compositions, methods and uses are provided for the administration of cell therapy of a defined composition at a specific dose that correlates with a high response rate and / or durability of response, and a low toxicity level and / or incidence. In some embodiments, the administered composition or dose is a flat and / or fixed dose of cells and / or one or more cells having a specific phenotype, e.g., a strictly flat dose, e.g., a number within a certain range and / or degree of variability or dispersion compared to a specific number or target number of such cells. In some embodiments, the administered composition or dose is a fixed dose of cells and / or one or more cells having a specific phenotype, e.g., a strictly flat dose, e.g., a number within a certain range and / or degree of variability or dispersion compared to a specific number or target number of such cells. + Cells and CD8 + Cells (e.g., CD4 + CAR + T cells: CD8 + CAR + In some embodiments, the ratio of CD4 T cells is 1:1, and / or is within a certain degree of variation from such ratio, e.g., within ±10%, e.g., within ±8%, e.g., the degree of variation or variance is ±10% or less, e.g., ±8% or less. + and CD8 + The cells are formulated and administered individually. In some embodiments, the administered cells exhibit consistent activity and / or function, such as cytokine production, apoptosis and / or expansion. In some embodiments, the provided compositions exhibit highly consistent and defined activity, as well as low variability between cells in the composition or between preparations, for example in terms of cell number, cell function, and / or cell activity. In some embodiments, the consistency in activity and / or function, such as low variability between preparations of the composition, allows for improved efficacy and / or safety. In some embodiments, administration of a defined composition resulted in low product variability and low toxicity, such as CRS or neurotoxicity, compared to administration of a cell composition with high heterogeneity. In some embodiments, a defined and consistent composition also exhibits consistent cell expansion. Such consistency can facilitate dose, identification of therapeutic windows, assessment of dose response, and identification of factors of interest that may correlate with safety or toxicity outcomes.

[0099] In some embodiments, in a certain cohort of subjects receiving a single infusion of a certain dose level, some cohort subjects can achieve an overall response rate (ORR, sometimes also known as objective response rate) of more than 80%, a complete response (CR) rate of more than 50% at 3 months. In some embodiments, subjects receiving the specified dose show improved safety outcomes. In some aspects, the rate of severe CRS or severe NT is low. In some embodiments, certain subject factors, such as certain biomarkers (e.g., TNF-α or IL-16), can be used to predict the risk of toxicity. In some embodiments, the provided embodiments can be used to achieve high response rates with low risk of toxicity.

[0100] In some embodiments, 25% or less, 20% or less, 15% or less, 10% or less, or 5% or less of subjects treated with the provided compositions, articles of manufacture, kits, methods and uses are administered an agent that ameliorates, treats, or prevents toxicity (e.g., tocilizumab and / or dexamethasone) prior to or following administration of the cell therapy. In some embodiments, subjects are not administered any preventative treatment prior to receiving the engineered cells (e.g., CAR-T cells).

[0101] In some embodiments, the methods provided provide advantages, for example, allow cell therapy to be administered on an outpatient basis. In some embodiments, cell therapy, for example, administration of a dose of T cells according to the embodiments provided, can be performed on an outpatient basis or does not require the subject to be admitted to a hospital, for example, hospital admission requiring an overnight stay. In some embodiments, such outpatient administration can allow increased access and reduced costs while maintaining high and sustained response rates with low toxicity. In some aspects, outpatient treatment can be advantageous for subjects who are already otherwise immunocompromised by previous treatment, for example, after lymphodepletion, and who are at higher risk of exposure during hospital stay or in hospitalization facilities. In some aspects, outpatient treatment also increases treatment options for subjects who may not have access to hospitalization, hospital facilities, or transplant centers, thereby expanding access to the treatment.

[0102] In some embodiments, the methods and uses result in or achieve a higher response rate and / or a more sustained response or efficacy, and / or a reduced risk of toxicity or other side effects that may be associated with cell therapy, such as neurotoxicity (NT) or cytokine release syndrome (CRS). In some aspects, the observations provided suggested a lower rate of severe NT (sNT) or severe CRS (sCRS), and a higher rate of patients without any toxicity, such as NT or CRS.

[0103] In some embodiments, at least at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% or more of subjects treated according to the provided methods and / or with the provided articles of manufacture or compositions achieve a complete response (CR). In some embodiments, at least 75%, at least 80%, or at least 90% of subjects treated according to the provided methods and / or with the provided articles of manufacture or compositions achieve an objective response (OR). In some embodiments, at least 35%, at least 45%, at least 50%, at least 55%, or at least 60% or more of subjects treated according to the provided methods and / or with the provided articles of manufacture or compositions achieve a CR or OR by 1 month, by 2 months, or by 3 months. In some embodiments, at least at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, or at least 75% or more of subjects who have failed previous treatment with a Bruton's tyrosine kinase inhibitor (BTKi) and venetoclax treated according to the provided methods and / or with the provided articles of manufacture or compositions achieve a complete response (CR). In some embodiments, at least 75%, at least 80%, or at least 90% of subjects who have failed previous treatment with a BTKi and venetoclax treated according to the provided methods and / or with the provided articles of manufacture or compositions achieve an objective response (OR). In some embodiments, at least 35%, at least 45%, at least 50%, at least 55%, or at least 60% or more of subjects who have failed previous treatment with a BTKi and venetoclax treated according to the provided methods and / or with the provided articles of manufacture or compositions achieve a CR or OR by 1 month, by 2 months, or by 3 months. In some embodiments, in subjects treated by the method, greater than 50%, greater than 60%, or greater than 70% had undetectable minimal residual disease (MRD) for at least 1 month, at least 2 months, at least 3 months, or at least 6 months after administration of a dose of cells.In some embodiments, in subjects who had failed previous treatment with BTKi and venetoclax treated according to the methods and / or with the provided articles of manufacture or compositions, greater than 50%, greater than 60%, or greater than 70% had undetectable minimal residual disease (MRD) for at least 1 month, at least 2 months, at least 3 months, or at least 6 months following administration of a dose of cells.

[0104] In some embodiments, up to 3 months after the start of administration of cell therapy, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, or at least 85% or more of subjects treated according to the provided methods and / or with the provided articles of manufacture or compositions remain in a response state, e.g., remain in a state of CR or OR and / or have undetectable MRD. In some embodiments, such a response, e.g., CR or OR, is sustainable for at least 3 months. In some embodiments, up to 3 months after the start of administration of cell therapy, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, or at least 85% or more of subjects who have failed previous treatment with BTKi and venetoclax treated according to the provided methods and / or with the provided articles of manufacture or compositions remain in a response state, e.g., remain in a state of CR or OR and / or have undetectable MRD. In some embodiments, such a response, e.g., CR or OR, is sustainable for at least 3 months.

[0105] In some embodiments, the resulting response observed in such subjects by treatment with the provided methods and / or with the provided articles of manufacture or compositions is associated with or results in a low risk of any toxicity or a low risk of severe toxicity in the majority of treated subjects. In some embodiments, more than or about 30%, 35%, 40%, 50%, 55%, or 60% or more of subjects treated according to the provided methods and / or with the provided articles of manufacture or compositions do not exhibit any grade of CRS or any grade of neurotoxicity (NT). In some embodiments, more than or about 50%, 60%, 70%, or 80% or more of subjects treated according to the provided methods and / or with the provided articles of manufacture or compositions do not exhibit severe CRS or grade 3 or higher CRS. In some embodiments, greater than or about 50%, 60%, 70%, or 80% or more of subjects treated according to the provided methods and / or with the provided articles of manufacture or compositions do not exhibit severe neurotoxicity or grade 3 or higher neurotoxicity, e.g., grade 4 or 5 neurotoxicity.

[0106] In some embodiments, the resulting responses observed in such subjects who have failed previous treatment with BTKi and venetoclax upon treatment according to the provided methods and / or with the provided articles of manufacture or compositions are associated with or result in a low risk of any toxicity or a low risk of serious toxicity in the majority of treated subjects. In some embodiments, greater than or about 30%, 35%, 40%, 50%, 55%, or 60% or more of subjects who have failed previous treatment with BTKi and venetoclax treated according to the provided methods and / or with the provided articles of manufacture or compositions do not exhibit any grade of CRS or any grade of neurotoxicity (NT). In some embodiments, greater than or about 50%, 60%, 70%, or 80% or more of subjects who have failed previous treatment with a BTKi and venetoclax treated according to the methods provided and / or with the articles of manufacture or compositions provided do not exhibit severe CRS or Grade 3 or higher CRS. In some embodiments, greater than or about 50%, 60%, 70%, or 80% or more of subjects who have failed previous treatment with a BTKi and venetoclax treated according to the methods provided and / or with the articles of manufacture or compositions provided do not exhibit severe neurotoxicity or Grade 3 or higher neurotoxicity, e.g., Grade 4 or 5 neurotoxicity.

[0107] In some embodiments, at least or about 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of subjects treated according to the methods and / or with the provided articles of manufacture or compositions do not exhibit early onset CRS or neurotoxicity and / or do not exhibit onset of CRS earlier than 1, 2, 3 or 4 days after initiation of administration. In some embodiments, at least or about 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of subjects treated according to the methods and / or with the provided articles of manufacture or compositions do not exhibit an onset of neurotoxicity earlier than 3, 4, 5, 6 or 7 days after the start of administration. In some aspects, the median onset of neurotoxicity among subjects treated according to the methods and / or with the provided articles of manufacture or compositions is at or after the median peak of CRS or the median time to resolution of CRS in subjects treated according to the methods. In some cases, the median onset of neurotoxicity among subjects treated according to the methods is greater than or greater than about 8, 9, 10 or 11 days.

[0108] In some embodiments, at least or about 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of subjects who have failed previous treatment with BTKi and venetoclax treated according to the methods and / or with the provided articles of manufacture or compositions do not exhibit early onset CRS or neurotoxicity and / or do not exhibit onset of CRS earlier than 1, 2, 3 or 4 days after initiation of dosing. In some embodiments, at least or about 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of subjects who have failed previous treatment with BTKi and venetoclax treated according to the methods and / or with the provided articles of manufacture or compositions do not exhibit an onset of neurotoxicity earlier than 3, 4, 5, 6 or 7 days after initiation of administration. In some aspects, the median onset of neurotoxicity among subjects who have failed previous treatment with BTKi and venetoclax treated according to the methods and / or with the provided articles of manufacture or compositions is at or after the median peak of CRS or the median time to resolution of CRS in subjects treated according to the methods. In some cases, the median onset of neurotoxicity among subjects who had failed previous treatment with a BTKi and venetoclax treated with the method is greater than or about 8, 9, 10, or 11 days.

[0109] In some embodiments, such results are greater than 2.5×10 7 or about 2.5 x 10 7 From 1.5 × 10 8 or about 1.5 x 10 8 Pieces, for example, about 5 x 10 7 From 1 × 10 8 or about 1 x 10 8total recombinant receptor-expressing T cells (e.g., CAR+ T cells), e.g., at a defined ratio as described herein, e.g., at a ratio of 1:1 or about 1:1, and / or CAR + A precise or uniform or fixed number of T cells or CD4 + CAR + T cells and / or CD8 + CAR + Certain types of CARs, such as T cells + CD4 T cells administered at a precise or uniform or fixed number, and / or any number of such cells within a specified degree of variance, e.g., + or - (plus or minus, in some cases denoted as ±) 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15% or less, as compared to such precise or uniform or fixed number. + and CD8 + In some embodiments, such a uniform or fixed number is observed after administration of a dose of T cells, including 2.5×10 7 Or about 2.5 x 10 7 Total CARs + T cells or CD8 + and / or CD4 + CAR + T cells, 5 x 10 7 Or about 5 x 10 7 Total CARs + T cells or CD8 + and / or CD4 + CAR + T cells, or 1 × 10 8 or about 1 x 10 8 Total CARs + T cells or CD8 + and / or CD4 + CAR + In some embodiments, the number of cells in the dose is 2.5×10 7 CARs + T cells (optional 1.25 x 10 7 CD4 + CAR + T cells and 1.25 × 10 7CD8 + CAR + T cells); in some embodiments, it comprises, consists of, or consists essentially of 5×10 7 CARs + T cells (optional 2.5 x 10 7 CD4 + CAR + T cells and 2.5 × 10 7 CD8 + CAR + T cells); in some embodiments, it comprises, consists of, or consists essentially of 1×10 8 CARs + T cells (optional 0.5 x 10 8 CD4 + CAR + T cells and 0.5 × 10 8 CD8 + CAR + In some aspects, the number of cells administered is within a certain degree of dispersion of such numbers in the above-mentioned embodiments, for example, within a range of plus or minus (±) 5, 6, 7, 8, 9, or 10%, for example, within a range of plus or minus 8%, compared to the number of such cells. In some aspects, the dose is within a certain degree of dispersion of such cells (for example, total CAR T cells). + T cells or CD8 + and / or CD4 + CAR + Within the range, a correlation (optionally a linear relationship) is observed between the number of cells (e.g., T cells) and one or more outcomes indicative of a therapeutic response or duration (e.g., remission, complete remission, and / or likelihood of achieving a particular duration of remission) and / or the duration of any of the foregoing. In some aspects, it is recognized that a higher dose of cells administered may result in a greater response without affecting or substantially affecting the incidence or risk of toxicity (e.g., CRS or neurotoxicity) or the degree of toxicity, e.g., severe CRS or severe neurotoxicity, in the subject.

[0110] In some aspects, subjects treated according to provided embodiments have adequate organ function. For example, in some aspects, subjects exhibit one or more of the following: serum creatinine ≦1.5×age-adjusted upper limit of normal (ULN) or estimated creatinine clearance (Cockcroft and Gault)>30 mL / min; alanine aminotransferase (ALT) ≦5×ULN and total bilirubin <2.0 mg / dL (or <3.0 mg / dL for subjects with Gilbert's syndrome or leukemic infiltration of the liver); ≦Common Terminology Criteria for Adverse Events (CTCAE) grade 1 dyspnea and saturated oxygen on room air (SaO 2 and / or adequate cardiac function, defined as a left ventricular ejection fraction (LVEF) of ≧40%, as assessed by an echocardiogram (ECHO) or multiple uptake gated acquisition (MUGA) scan performed within 30 days prior to evaluating the subject for administration of the engineered cell composition.

[0111] In some aspects, the provided methods provide for high or particular response rates (e.g., response rates in a population evaluated after a certain period of time after administration, e.g., 1 month or 3 months), e.g., ORR of 75% or more, 80% or more, 85% or more, or about 75% or more, 80% or more, 85% or more (e.g., 1 month ORR or 3 month ORR), and ORR of 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, CR rates (e.g., 1-month CR rate or 3-month CR rate) of 60% or more, 65% or more, 70% or more, 71% or more, 72% or more, 73% or more, 74% or more, or about 75% or more, or about 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 71% or more, 72% or more, 73% or more, 74% or more, or about 75% or more can be achieved. In some embodiments, such response rates and durability are only obtained after a single administration or dose of such therapy. Treatment of such subjects with the provided methods and / or with the provided articles of manufacture or compositions also results in subjects that, in some embodiments, achieve high response rates but do not exhibit higher incidences of toxicity, such as neurotoxicity or CRS, even with higher cell doses.

[0112] Thus, in some embodiments, the provided methods, articles of manufacture, and / or compositions can provide advantages over other available methods, solutions, or approaches for treatment, e.g., adoptive cell therapy. In particular, some of the provided embodiments provide advantages to subjects with high-risk CLL by achieving a high rate of durable responses while reducing the incidence of toxicity or side effects.

[0113] A. Treatment Method Methods of treatment involving administration of engineered cells, such as engineered T cells, or compositions comprising engineered cells are provided herein. Methods and uses of engineered cells (e.g., T cells) and / or compositions thereof are also provided, including methods for the treatment of subjects with diseases or conditions, such as leukemia or lymphoma, e.g., chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL), involving administration of engineered cells and / or compositions thereof. In some embodiments, the methods and uses provided can achieve improved response and / or more sustained response or efficacy and / or reduced risk of toxicity or other side effects, for example, in a particular group of treated subjects, compared to certain alternative methods. In some aspects, methods of administering engineered cells, such as engineered T cells, or compositions comprising engineered cells to a subject, e.g., a subject with a disease or disorder, are also provided. In some aspects, the use of engineered cells, such as engineered T cells, or compositions comprising engineered cells for the treatment of a disease or disorder is also provided. In some aspects, the use of engineered cells, such as engineered T cells, or compositions comprising engineered cells, for the manufacture of medicaments for the treatment of disease or disorder is also provided.In some aspects, the method of administering engineered cells, such as engineered T cells, or compositions comprising engineered cells, for use in the treatment of disease or disorder or for administration to subjects with disease or disorder is also provided.In some aspects, the use of engineered cells, such as engineered T cells, or compositions comprising engineered cells, is according to any of the methods described herein.

[0114] The engineered cells expressing recombinant receptors such as chimeric antigen receptors (CARs) described herein or compositions comprising the same are useful in a variety of therapeutic, diagnostic and prophylactic applications. For example, the engineered cells or compositions comprising the engineered cells are useful for treating a variety of diseases and disorders in a subject. Such methods and uses include therapeutic methods and uses involving administration of the engineered cells or compositions comprising the same to a subject having a disease, condition, or disorder, such as, for example, a tumor or cancer. In some embodiments, the engineered cells or compositions comprising the same are administered in an amount effective to effect treatment of the disease or disorder. Uses include the use of the engineered cells or compositions in such methods and treatments, and in the preparation of medicaments for carrying out such therapeutic methods. In some embodiments, the engineered cells or compositions comprising the engineered cells are for use in treating a variety of diseases and disorders in a subject, for example, according to a therapeutic method. In some embodiments, the method is carried out by administering the engineered cells or compositions comprising the same to a subject having or suspected of having a disease or condition. In some embodiments, the method thereby treats the disease or condition or disorder in the subject.

[0115] General methods for administration of cells for adoptive cell therapy are known and can be used in connection with the provided methods and compositions.For example, methods of adoptive T cell therapy are described, for example, in U.S. Patent Application Publication No. 2003 / 0170238 to Gruenberg et al.; U.S. Patent No. 4,690,915 to Rosenberg; Rosenberg (2011) Nat Rev Clin Oncol. 8(10):577-85).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; Davila et al. (2013) PLoS ONE 8(4): e61338.

[0116] The disease or condition to be treated may be any in which the expression of an antigen is associated with and / or involved in the pathogenesis of the disease, condition or disorder, e.g., causes, exacerbates, or otherwise participates in such disease, condition, or disorder. Exemplary diseases and conditions may include diseases or conditions associated with malignant tumors or cell transformation (e.g., cancer), autoimmune or inflammatory diseases, or infectious diseases, e.g., caused by bacterial, viral, or other pathogens. Exemplary antigens, including antigens associated with various diseases and conditions that can be treated, are described above. In certain embodiments, the chimeric antigen receptor or transgenic TCR specifically binds to an antigen associated with the disease or condition.

[0117] Among the diseases, conditions and disorders are tumors, including solid tumors, hematological malignancies and melanomas, as well as localized and metastatic tumors, infectious diseases, such as infections with viral or other pathogens, such as HIV, HCV, HBV, CMV, HPV, and parasitic diseases, as well as autoimmune and inflammatory diseases.In some embodiments, the disease, disorder or condition is tumor, cancer, malignancy, neoplasm, or other proliferative disease or disorder.Such diseases for treatment by the methods provided herein include, but are not limited to, leukemia, lymphoma, such as chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL).

[0118] In some embodiments, the Eastern Cooperative Oncology Group (ECOG) performance status index can be used to evaluate or select subjects for treatment, e.g., subjects who have had poor outcomes with previous therapies (see, e.g., Oken et al. (1982) Am J Clin Oncol. 5:649-655). The performance status of the ECOG scale describes the level of functionality of a patient in terms of their ability to care for themselves, their daily activities, and their physical abilities (e.g., walking, working, etc.). In some embodiments, an ECOG performance status of 0 indicates that the patient can carry out normal activities. In some aspects, a subject with an ECOG performance status of 1 shows some limitations in physical activities, but the subject is fully ambulatory. In some aspects, a patient with an ECOG performance status of 2 is more than 50% ambulatory. In some cases, a subject with an ECOG performance status of 2 is also able to care for themselves; see, e.g., Sorensen et al., (1993) Br J Cancer 67(4) 773-775. The criteria reflecting ECOG performance status are listed in Table 1 below:

[0119] (Table 1) ECOG performance status criteria TIFF0007652692000001.tif47167

[0120] In some embodiments, the antigen targeted by the receptor (e.g., CAR) includes an antigen associated with B cell malignancies, such as any of several known B cell markers. In some embodiments, the antigen is or comprises CD20, CD19, CD22, ROR1, CD45, CD21, CD5, CD33, Igκ, Igλ, CD79a, CD79b, or CD30. In some embodiments, the antigen is CD19.

[0121] In some embodiments, cell therapy, such as adoptive T cell therapy, is performed by autologous transfer, in which cells are isolated and / or otherwise prepared from the subject who is to receive cell therapy or from a sample derived from such a subject.Thus, in some aspects, cells are derived from a subject, such as a patient, in need of treatment, and the cells are administered to the same subject after isolation and processing.

[0122] In some embodiments, cell therapy, such as adoptive T cell therapy, is carried out by allogeneic transfer, in which cells are isolated and / or otherwise prepared from a subject other than the subject who is going to receive cell therapy or the subject who will ultimately receive cell therapy, such as a first subject.In such embodiments, the cells are then administered to a different subject of the same species, such as a second subject.In some embodiments, the first subject and the second subject are genetically identical.In some embodiments, the first subject and the second subject are genetically similar.In some embodiments, the second subject expresses the same HLA class or supertype as the first subject.

[0123] The cells can be administered by any suitable means, for example, by bolus injection, by injection, for example, intravenous or subcutaneous injection, intraocular injection, periocular injection, subretinal injection, intravitreal injection, transseptal injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjectval injection, subconjuntival injection, subtenon injection, retrobulbar injection, peribulbar injection, or posterior juxtascleral delivery. In some embodiments, they are administered parenterally, intrapulmonary, and intranasally, as well as intralesional administration when local treatment is desired. Parenteral injection includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In some embodiments, a certain dose is administered by a single bolus administration of cells. In some embodiments, it is administered by multiple boluses of cells or by continuous infusion administration of cells, for example, over a period of 3 days or less. In some embodiments, administration of the cell dose or any additional therapy, e.g., lymphodepleting therapy, interventional therapy, and / or combination therapy, is via outpatient delivery.

[0124] For the prevention or treatment of disease, the appropriate dosage can be determined by the type of disease being treated, the type of cells or recombinant receptor, the severity and course of the disease, whether the cells are administered for prophylactic or therapeutic purposes, previous treatments, the subject's medical history and response to the cells, and the judgment of the attending physician. The compositions and cells, in some embodiments, are suitably administered to the subject at one time or over a series of treatments.

[0125] In some aspects, the methods include administration of a chemotherapeutic agent, such as a conditioning chemotherapeutic agent.

[0126] Preconditioning a subject with immunodepleting (eg, lymphodepleting) therapy in some aspects can improve the efficacy of adoptive cellular therapy (ACT).

[0127] Thus, in some embodiments, the method comprises administering a preconditioning agent, such as a lymphodepleting or chemotherapeutic agent, such as cyclophosphamide, fludarabine, or a combination thereof, to the subject before the start of cell therapy.For example, the subject may administer the preconditioning agent at least 2 days before the start of cell therapy, such as at least 3 days, 4 days, 5 days, 6 days, or 7 days before.In some embodiments, the subject is administered the preconditioning agent within 7 days before the start of cell therapy, such as within 6 days, 5 days, 4 days, 3 days, or 2 days before the start of cell therapy.

[0128] In some embodiments, the subject is preconditioned with cyclophosphamide at a dose of 20 mg / kg to 100 mg / kg or about 20 mg / kg to 100 mg / kg, for example, 40 mg / kg to 80 mg / kg or about 40 mg / kg to 80 mg / kg. In some aspects, the subject is preconditioned with 60 mg / kg or about 60 mg / kg of cyclophosphamide. In some embodiments, the cyclophosphamide may be administered in a single dose or in multiple doses, for example, given daily, every other day, or every third day. In some embodiments, the cyclophosphamide is administered once a day for one or two days. In some embodiments, when the lymphodepleting agent includes cyclophosphamide, the subject is preconditioned with cyclophosphamide at a dose of 100 mg / m 2 ~500 mg / m 2 or about 100 mg / m 2 ~500 mg / m 2 , e.g., 200 mg / m 2 ~400 mg / m 2 or about 200 mg / m 2 ~400 mg / m 2 , or 250 mg / m 2 ~350 mg / m 2 or about 250 mg / m 2 ~350 mg / m 2 In some examples, the subject is administered cyclophosphamide at a dose of about 300 mg / m 2of cyclophosphamide is administered. In some embodiments, cyclophosphamide may be administered in a single dose or in multiple doses, e.g., given daily, every other day, or every third day. In some embodiments, cyclophosphamide is administered daily, e.g., for 1-5 days, e.g., for 3-5 days. In some examples, the subject is administered about 300 mg / m daily for 3 days prior to the start of cell therapy. 2 of cyclophosphamide is administered.

[0129] In some embodiments, when the lymphodepletion agent comprises fludarabine, the subject is administered 1 mg / m 2 ~100 mg / m 2 or about 1 mg / m 2 ~100 mg / m 2 , e.g., 10 mg / m 2 ~75 mg / m 2 or about 10 mg / m 2 ~75 mg / m 2 , 15 mg / m 2 ~50 mg / m 2 or about 15 mg / m 2 ~50 mg / m 2 , 20 mg / m 2 ~40 mg / m 2 or about 20 mg / m 2 ~40 mg / m 2 , or 24 mg / m 2 ~35 mg / m 2 or about 24 mg / m 2 ~35 mg / m 2 In some cases, subjects are administered fludarabine at a dose of about 30 mg / m 2 of fludarabine is administered. In some embodiments, fludarabine may be administered in a single dose or in multiple doses, e.g., given daily, every other day, or every third day. In some embodiments, fludarabine is administered daily, e.g., for 1-5 days, e.g., for 3-5 days. In some examples, the subject is administered about 30 mg / m2 daily for 3 days prior to the start of cell therapy. 2 of fludarabine is administered.

[0130] In some embodiments, the lymphodepleting agent comprises a combination of agents, such as a combination of cyclophosphamide and fludarabine. Thus, the combination of agents may comprise cyclophosphamide at any dose or administration schedule, such as cyclophosphamide as described above, and fludarabine at any dose or administration schedule, such as fludarabine as described above. For example, in some aspects, the subject is administered 60 mg / kg (~2 g / m) of cyclophosphamide prior to the first or subsequent dose. 2 ) of cyclophosphamide and 25 mg / m 2 Three to five doses of fludarabine are administered.

[0131] After administration of the cells, the biological activity of the engineered cell population in some embodiments is measured, for example, by any of several known methods. The parameters evaluated include the specific binding of engineered or natural T cells or other immune cells to antigens in vivo, for example, by imaging, or ex vivo, for example, by ELISA or flow cytometry. In certain embodiments, the ability of the engineered cells to destroy target cells can be measured using any suitable known method, such as the cytotoxicity assay described in Kochenderfer et al., J. Immunotherapy, 32(7): 689-702 (2009), and Herman et al. J. Immunological Methods, 285(1): 25-40 (2004). In certain embodiments, the biological activity of the cells is measured by assaying the expression and / or secretion of one or more cytokines, for example, CD107a, IFNγ, IL-2, and TNF.

[0132] In certain embodiments, the engineered cells are further modified in any number of ways to increase their therapeutic or prophylactic efficacy.For example, the engineered CAR or TCR expressed by the population can be conjugated to a targeting moiety, either directly or indirectly through a linker.Methods for conjugating a compound, such as a CAR or TCR, to a targeting moiety are known.See, for example, Wadwa et al., J. Drug Targeting 3:111(1995) and U.S. Patent No. 5,087,616.In some embodiments, the cells are administered as part of a combination treatment, for example, simultaneously with another therapeutic intervention, such as an antibody or engineered cell or receptor or agent, such as a cytotoxic agent or therapeutic agent, or sequentially in any order therewith.The cells in some embodiments are co-administered simultaneously or sequentially in any order with one or more additional therapeutic agents or in association with another therapeutic intervention. In some situations, the cells are co-administered with another therapy close enough in time that the cell population enhances the effect of one or more additional therapeutic agents, or vice versa.In some embodiments, the cells are administered before one or more additional therapeutic agents.In some embodiments, the cells are administered after one or more additional therapeutic agents.In some embodiments, the one or more additional agents include cytokines, such as IL-2, for example, to enhance persistence.

[0133] B. Administration In some embodiments, a dose of cells is administered to a subject according to the provided method and / or by the provided article of manufacture or composition.In some embodiments, the size or timing of the dose is determined as a function of the specific disease or condition in the subject.In some cases, the size or timing of the dose for a specific disease can be empirically determined in light of the provided description.

[0134] In certain embodiments, individual populations of cells, or subtypes of cells, may be present in a range of about 1 million to about 100 billion cells, and / or in an amount of such cells per kilogram of body weight, e.g., about 1 million to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or a range defined by any two of the foregoing values), e.g., about 10 million to about 100 billion cells (e.g., about 20 million cells, about 30 million cells, about 40 million cells, about 60 million cells, about 70 million cells, about 80 million cells, or a range defined by any two of the foregoing values). The subject may be administered about 100 million cells, about 90 million cells, about 10 billion cells, about 25 billion cells, about 50 billion cells, about 75 billion cells, about 90 billion cells, or a range defined by any two of the foregoing values), and in some cases, about 100 million cells to about 50 billion cells (e.g., about 120 million cells, about 250 million cells, about 350 million cells, about 450 million cells, about 650 million cells, about 800 million cells, about 900 million cells, about 3 billion cells, about 30 billion cells, about 45 billion cells), or any value between these ranges and / or any value between these ranges per kilogram of body weight. Dosages may vary depending on the specific nature of the disease or disorder and / or the patient and / or other treatments. In some embodiments, such values ​​refer to the number of recombinant receptor-expressing cells; in other embodiments, they refer to the number of T cells or PBMCs or total cells administered. In some embodiments, the number of cells is the number of such cells that are viable cells.

[0135] In some embodiments, the dose of cells is a flat dose of cells or a fixed dose of cells, such that the cell dose is not related to or based on the subject's body surface area or weight.

[0136] In some embodiments, the dose of genetically engineered cells is 1×10 5 Or about 1 x 10 5 From, 5 × 108 Or about 5 x 10 8 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 2.5 × 10 8 Or about 2.5 x 10 8 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 1 × 10 8 Or about 1 x 10 8 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From, 5 × 10 7 Or about 5 x 10 7 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 2.5 × 10 7 Or about 2.5 x 10 7 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 1 × 10 7 Or about 1 x 10 7 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From, 5 × 10 6 Or about 5 x 10 6 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 2.5 × 10 6 Or about 2.5 x 10 6 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 1 × 10 6 Or about 1 x 10 6 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From, 5 × 10 8 Or about 5 x 10 8 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From 2.5 × 10 8 Or about 2.5 x 10 8 Total CAR-expressing T cells, 1 x 106 Or about 1 x 10 6 From 1 × 10 8 Or about 1 x 10 8 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From, 5 × 10 7 Or about 5 x 10 7 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From 2.5 × 10 7 Or about 2.5 x 10 7 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From 1 × 10 7 Or about 1 x 10 7 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From, 5 × 10 6 Or about 5 x 10 6 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From 2.5 × 10 6 Or about 2.5 x 10 6 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6 From, 5 × 10 8 Or about 5 x 10 8 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6 From 2.5 × 10 8 Or about 2.5 x 10 8 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6 From 1 × 10 8 Or about 1 x 10 8 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6 From, 5 × 10 7 Or about 5 x 10 7 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6From 2.5 × 10 7 Or about 2.5 x 10 7 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6 From 1 × 10 7 Or about 1 x 10 7 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6 From, 5 × 10 6 Or about 5 x 10 6 Total CAR-expressing T cells, 5 x 10 6 Or about 5 x 10 6 From, 5 × 10 8 Or about 5 x 10 8 Total CAR-expressing T cells, 5 x 10 6 Or about 5 x 10 6 From 2.5 × 10 8 Or about 2.5 x 10 8 Total CAR-expressing T cells, 5 x 10 6 Or about 5 x 10 6 From 1 × 10 8 Or about 1 x 10 8 Total CAR-expressing T cells, 5 x 10 6 Or about 5 x 10 6 From, 5 × 10 7 Or about 5 x 10 7 Total CAR-expressing T cells, 5 x 10 6 Or about 5 x 10 6 From 2.5 × 10 7 Or about 2.5 x 10 7 Total CAR-expressing T cells, 5 x 10 6 Or about 5 x 10 6 From 1 × 10 7 Or about 1 x 10 7 Total CAR-expressing T cells, 1 x 10 7 Or about 1 x 10 7 From, 5 × 10 8 Or about 5 x 10 8 Total CAR-expressing T cells, 1 x 10 7 Or about 1 x 10 7 From 2.5 × 10 8 Or about 2.5 x 10 8Total CAR-expressing T cells, 1 x 10 7 Or about 1 x 10 7 From 1 × 10 8 Or about 1 x 10 8 Total CAR-expressing T cells, 1 x 10 7 Or about 1 x 10 7 From, 5 × 10 7 Or about 5 x 10 7 Total CAR-expressing T cells, 1 x 10 7 Or about 1 x 10 7 From 2.5 × 10 7 Or about 2.5 x 10 7 Total CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 From, 5 × 10 8 Or about 5 x 10 8 Total CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 From 2.5 × 10 8 Or about 2.5 x 10 8 Total CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 From 1 × 10 8 Or about 1 x 10 8 Total CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 From, 5 × 10 7 Or about 5 x 10 7 Total CAR-expressing T cells, 5 x 10 7 Or about 5 x 10 7 From, 5 × 10 8 Or about 5 x 10 8 Total CAR-expressing T cells, 5 x 10 7 Or about 5 x 10 7 From 2.5 × 10 8 Or about 2.5 x 10 8 Total CAR-expressing T cells, 5 x 10 7 Or about 5 x 10 7 From 1 × 10 8 Or about 1 x 10 8 Total CAR-expressing T cells, 1 x 10 8 Or about 1 x 108 From, 5 × 10 8 Or about 5 x 10 8 Total CAR-expressing T cells, 1 x 10 8 Or about 1 x 10 8 From 2.5 × 10 8 Or about 2.5 x 10 8 Total CAR-expressing T cells, 2.5 x 10 8 Or about 2.5 x 10 8 From, 5 × 10 8 Or about 5 x 10 8 In some embodiments, the dose of engineered cells comprises 2.5×10 total CAR-expressing T cells. 7 Or about 2.5 x 10 7 From 1.5 × 10 8 Or about 1.5 x 10 8 Total CAR-expressing T cells, e.g. 5 x 10 7 Or about 5 x 10 7 From 1 × 10 8 Or about 1 x 10 8 The total number of CAR-expressing T cells. In some embodiments, the number of cells is the number of such cells that are viable cells, e.g., viable T cells. In some embodiments, the number of cells is the number of such cells that are live cells.

[0137] In some embodiments, a dose of genetically engineered cells is greater than or equal to 2.5×10 7 or about 2.5 x 10 7 From 1.5 × 10 8 or about 1.5 x 10 8 Total CAR-expressing T cells, e.g. 5 x 10 7 ~1×10 8 In some embodiments, a dose of engineered cells comprises at least 2.5×10 total CAR-expressing T cells. 7 Or at least about 2.5 × 10 7 CAR-expressing cells, at least 5 x 10 7 Or at least about 5×10 7 CAR-expressing cells, or at least 1 x 10 8 Or at least about 1 × 10 8In some embodiments, a dose of T cells comprises 2.5×10 CAR-expressing cells. 7 or about 2.5 x 10 7 In some embodiments, a dose of T cells comprises 1×10 8 pcs or approx. 1×10 8 In some embodiments, a dose of T cells comprises 5×10 7 Pieces or about 5 x 10 7 In some embodiments, the number of cells is the number of such cells that are viable cells.

[0138] In some embodiments, the number is CD3 + , CD8 + , or CD4+ and CD8+ cells, and in some cases recombinant receptor expression (e.g., CAR + ) refers to the total number of cells. In some embodiments, the number of cells is the number of such cells that are viable cells.

[0139] In some embodiments, the cell therapy comprises a cell number between 1×10 5 Or about 1 x 10 5 From, 5 × 10 8 Or about 5 x 10 8 CD3 + , CD8 + Or CD4 + and CD8 + Total T cells or CD3 + , CD8 + Or CD4 + and CD8 + Recombinant receptor (e.g., CAR+) expressing cells, 5 × 10 5 Or about 5 x 10 5 From 1 × 10 7 Or about 1 x 10 7 CD3 + , CD8 + Or CD4 + and CD8 + Total T cells or CD3 + , CD8 + Or CD4 +and CD8 + Recombinant receptor (e.g., CAR+) expressing cells, or 1 × 10 6 Or about 1 x 10 6 From 1 × 10 7 Or about 1 x 10 7 CD3 + , CD8 + Or CD4 + and CD8 + Total T cells or CD3 + , CD8 + Or CD4 + and CD8 + In some embodiments, the cell therapy comprises administering a dose comprising recombinant receptor (e.g., CAR+) expressing cells. In some embodiments, the cell therapy comprises administering a dose comprising recombinant receptor (e.g., CAR+) expressing cells, the number of cells being 1×10, inclusive. 5 Or about 1 x 10 5 From, 5 × 10 8 Or about 5 x 10 8 Total CD3 + / CAR + , CD8 + / CAR + Or CD4 + / CD8 + / CAR + cells, 5 x 10 5 Or about 5 x 10 5 From 1 × 10 7 Or about 1 x 10 7 Total CD3 + / CAR + , CD8 + / CAR + Or CD4 + / CD8 + / CAR + cells, or 1 x 10 6 Or about 1 x 10 6 From 1 × 10 7 Or about 1 x 10 7 Total CD3 + / CAR + , CD8 + / CAR + Or CD4 + / CD8 + / CAR +It includes administering a dose comprising cells. In some embodiments, the number of cells is the number of such cells that are viable cells.

[0140] In some embodiments, a dose of genetically engineered cells is greater than or equal to 2.5×10 7 Or about 2.5 x 10 7 From 1.5 × 10 8 Total CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + T cells, e.g., 5 x 10 7 ~1×10 8 Total CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + In some embodiments, the dose of engineered cells comprises at least 2.5×10 T cells. 7 Or at least about 2.5 × 10 7 CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + T cells, at least 5 × 10 7 Or at least about 5×10 7 CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + T cells, or at least 1 × 10 8 Or at least about 1 × 10 8 CD3 + / CAR + , CD8 + / CAR + , or CD4+ / CD8 + / CAR + In some embodiments, the dose of engineered cells comprises 2.5×10 7 Or about 2.5 x 10 7 CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + T cells, 5 x 10 7 Or about 5 x 10 7 CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + T cells, or 1 × 10 8 Or about 1 x 10 8 CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + In some embodiments, the number of cells is the number of such cells that are viable cells.

[0141] In some embodiments, a dose of T cells comprises 5×10 7 Or about 5 x 10 7 recombinant receptor (e.g., CAR)-expressing T cells, or 2.5 x 10 7 Or about 2.5 x 10 7 Recombinant receptor (e.g., CAR)-expressing CD8 + In some embodiments, a dose of T cells comprises 1×10 8 Or about 1 x 10 8 recombinant receptor (e.g., CAR)-expressing T cells, or 5 x 10 7 Or about 5 x 10 7 Recombinant receptor (e.g., CAR)-expressing CD8 +In some embodiments, a dose of T cells comprises 1.5×10 8 Or about 1.5 x 10 8 recombinant receptor (e.g., CAR)-expressing T cells, or 0.75 x 10 8 Or about 0.75 x 10 8 Recombinant receptor (e.g., CAR)-expressing CD8 + In some embodiments, the number of cells is the number of such cells that are viable cells.

[0142] In some embodiments, the dose of T cells comprises CD4 + T cells, CD8 + T cells, or CD4 + and CD8 + In some embodiments, for example, when the subject is a human, the subject may be a CD4 + and CD8 + A dose of CD8 + T cells are approximately 2.5 × 10 7 ~1×10 8 Total recombinant receptor (e.g., CAR)-expressing CD8 + cells, or CD4 + Cells vs. CD8 + The present invention includes a fraction thereof, optionally present in a ratio of 1:1 or about 1:1 to T cells.

[0143] In some embodiments, a patient is administered multiple doses, and each or all of the doses can be within any of the aforementioned values. In some embodiments, a dose of cells is greater than 1×10 5 Or about 1 x 10 5 From, 5 × 10 8 Or about 5 x 10 8 1 x 10 total recombinant receptor (e.g., CAR)-expressing T cells or total T cells 5 Or about 1 x 10 5 From 1.5 × 10 8 Or about 1.5 x 10 8 1 x 10 total recombinant receptor (e.g., CAR)-expressing T cells or total T cells 5 Or about 1 x 10 5From 1 × 10 8 Or about 1 x 10 8 5 x 10 total recombinant receptor (e.g., CAR)-expressing T cells or total T cells 5 Or about 5 x 10 5 From 1 × 10 7 Or about 1 x 10 7 10 total recombinant receptor (e.g., CAR)-expressing T cells or total T cells, or 1 x 10 6 Or about 1 x 10 6 From 1 × 10 7 Or about 1 x 10 7 The present invention includes administration of total recombinant receptor (e.g., CAR)-expressing T cells or total T cells (each inclusive).

[0144] In some embodiments, the dose of cells is a flat dose or a fixed dose of cells, such that the dose of cells is not related to or based on the subject's body surface area or weight.

[0145] In some embodiments, a dose of genetically engineered cells is greater than or equal to 1×10 5 Or about 1 x 10 5 From 5×10 8 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 2.5 × 10 8 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 1×10 8 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 5×10 7 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 2.5 × 10 7 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 1×10 7 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 5×10 6Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 2.5 × 10 6 Total CAR-expressing T cells, 1 x 10 5 Or about 1 x 10 5 From 1×10 6 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From 5×10 8 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From 2.5 × 10 8 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From 1×10 8 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From 5×10 7 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From 2.5 × 10 7 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From 1×10 7 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From 5×10 6 Total CAR-expressing T cells, 1 x 10 6 Or about 1 x 10 6 From 2.5 × 10 6 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6 From 5×10 8 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6 From 2.5 × 10 8 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6 From 1×10 8 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6From 5×10 7 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6 From 2.5 × 10 7 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6 From 1×10 7 Total CAR-expressing T cells, 2.5 x 10 6 Or about 2.5 x 10 6 From 5×10 6 Total CAR-expressing T cells, 5 x 10 6 Or about 5 x 10 6 From 5×10 8 Total CAR-expressing T cells, 5 x 10 6 Or about 5 x 10 6 From 2.5 × 10 8 Total CAR-expressing T cells, 5 x 10 6 Or about 5 x 10 6 From 1×10 8 Total CAR-expressing T cells, 5 x 10 6 Or about 5 x 10 6 From 5×10 7 Total CAR-expressing T cells, 5 x 10 6 Or about 5 x 10 6 From 2.5 × 10 7 Total CAR-expressing T cells, 5 x 10 6 Or about 5 x 10 6 From 1×10 7 Total CAR-expressing T cells, 1 x 10 7 Or about 1 x 10 7 From 5×10 8 Total CAR-expressing T cells, 1 x 10 7 Or about 1 x 10 7 From 2.5 × 10 8 Total CAR-expressing T cells, 1 x 10 7 Or about 1 x 10 7 From 1×10 8 Total CAR-expressing T cells, 1 x 10 7 Or about 1 x 10 7 From 5×10 7 Total CAR-expressing T cells, 1 x 10 7 Or about 1 x 107 From 2.5 × 10 7 Total CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 From 5×10 8 Total CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 From 2.5 × 10 8 Total CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 From 1×10 8 Total CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 From 5×10 7 Total CAR-expressing T cells, 5 x 10 7 Or about 5 x 10 7 From 5×10 8 Total CAR-expressing T cells, 5 x 10 7 Or about 5 x 10 7 From 2.5 × 10 8 Total CAR-expressing T cells, 5 x 10 7 Or about 5 x 10 7 From 1×10 8 Total CAR-expressing T cells, 1 x 10 8 Or about 1 x 10 8 From 5×10 8 Total CAR-expressing T cells, 1 x 10 8 Or about 1 x 10 8 From 2.5 × 10 8 total CAR-expressing T cells, or 2.5 x 10 8 Or about 2.5 x 10 8 From 5×10 8 In some embodiments, the number of cells is the number of such cells that are viable cells.

[0146] In some embodiments, a dose of genetically engineered cells is greater than or equal to 2.5×10 7 or about 2.5 x 10 7 From 1.5 × 10 8 or about 1.5 x 10 8 Total CAR-expressing T cells, e.g. 5 x 107 ~1×10 8 In some embodiments, a dose of engineered cells comprises at least 2.5×10 total CAR-expressing T cells. 7 Or at least about 2.5 × 10 7 CAR-expressing cells, at least 5 x 10 7 Or at least about 5×10 7 CAR-expressing cells, or at least 1 x 10 8 Or at least about 1 × 10 8 In some embodiments, a dose of T cells comprises 2.5×10 CAR-expressing cells. 7 or about 2.5 x 10 7 In some embodiments, a dose of T cells comprises 1×10 8 or about 1 x 10 8 In some embodiments, a dose of T cells comprises 5×10 7 or about 5 x 10 7 In some embodiments, the number of cells is the number of such cells that are viable cells.

[0147] In some embodiments, the number is CD3 + , CD8 + , or CD4 + And CD8 + In relation to the total number of cells, in some cases, recombinant receptor expression (e.g., CAR + ) also refers to the total number of cells. In some embodiments, the number of cells is the number of such cells that are viable cells.

[0148] In some aspects, the cell therapy comprises at least 1×10 5 Or about 1 x 10 5 From, 5 × 10 8 Or about 5 x 10 8 CD3 + , CD8 + , or CD4 + and CD8 + Total T cells, or CD3 + , CD8 + , or CD4 + and CD8+ Recombinant receptors (e.g., CAR + ) expressing cells, 5 × 10 5 Or about 5 x 10 5 From 1 × 10 7 Or about 1 x 10 7 CD3 + , CD8 + , or CD4 + and CD8 + Total T cells, or CD3 + , CD8 + , or CD4 + and CD8 + Recombinant receptors (e.g., CAR + ) expressing cells, or 1 × 10 6 Or about 1 x 10 6 From 1 × 10 7 Or about 1 x 10 7 CD3 + , CD8 + , or CD4 + and CD8 + Total T cells, or CD3 + , CD8 + , or CD4 + and CD8 + Recombinant receptors (e.g., CAR + In some embodiments, the cell therapy comprises administering a dose comprising a number of cells of 1×10 5 Or about 1 x 10 5 From, 5 × 10 8 Or about 5 x 10 8 Total CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + cells, 5 x 10 5 Or about 5 x 10 5 From 1 × 10 7 Or about 1 x 10 7 Total CD3 + / CAR + , CD8 + / CAR + , or CD4+ / CD8 + / CAR + cells, or 1 x 10 6 Or about 1 x 10 6 From 1 × 10 7 Or about 1 x 10 7 Total CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + It includes administering a dose that includes a number of cells of cells (each of the endpoints inclusive). In some embodiments, the number of cells is the number of such cells that are viable cells.

[0149] In some embodiments, a dose of genetically engineered cells is greater than or equal to 2.5×10 7 Or about 2.5 x 10 7 From 1.5 × 10 8 Total CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + T cells, e.g., 5 x 10 7 ~1×10 8 Total CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + In some embodiments, the dose of engineered cells comprises at least 2.5×10 T cells. 7 Or at least about 2.5 × 10 7 CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + T cells, at least 5 × 10 7 Or at least about 5×107 CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + T cells, or at least 1 × 10 8 Or at least about 1 × 10 8 CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + In some embodiments, the dose of engineered cells comprises 2.5×10 7 Or about 2.5 x 10 7 CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + T cells, 5 x 10 7 Or about 5 x 10 7 CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + T cells, or 1 × 10 8 Or about 1 x 10 8 CD3 + / CAR + , CD8 + / CAR + , or CD4 + / CD8 + / CAR + In some embodiments, the number of cells is the number of such cells that are viable cells.

[0150] In some embodiments, a dose of T cells comprises 5×10 7 Or about 5 x 10 7recombinant receptor (e.g., CAR)-expressing T cells, or 2.5 x 10 7 Or about 2.5 x 10 7 Recombinant receptor (e.g., CAR)-expressing CD8 + In some embodiments, a dose of T cells comprises 1×10 8 Or about 1 x 10 8 recombinant receptor (e.g., CAR)-expressing T cells, or 5 x 10 7 Or about 5 x 10 7 Recombinant receptor (e.g., CAR)-expressing CD8 + In some embodiments, a dose of T cells comprises 1.5×10 8 Or about 1.5 x 10 8 recombinant receptor (e.g., CAR)-expressing T cells, or 0.75 x 10 8 Or about 0.75 x 10 8 Recombinant receptor (e.g., CAR)-expressing CD8 + In some embodiments, the number of cells is the number of such cells that are viable cells.

[0151] In some embodiments, a patient is administered multiple doses, and each or all of the doses can be within any of the aforementioned values. In some embodiments, a dose of cells is greater than 1×10 5 Or about 1 x 10 5 From, 5 × 10 8 Or about 5 x 10 8 1 x 10 total recombinant receptor (e.g., CAR)-expressing T cells or total T cells 5 Or about 1 x 10 5 From 1.5 × 10 8 Or about 1.5 x 10 8 1 x 10 total recombinant receptor (e.g., CAR)-expressing T cells or total T cells 5 Or about 1 x 10 5 From 1 × 10 8 Or about 1 x 10 8 5 x 10 total recombinant receptor (e.g., CAR)-expressing T cells or total T cells 5 Or about 5 x 10 5 From 1 × 10 7Or about 1 x 10 7 10 total recombinant receptor (e.g., CAR)-expressing T cells or total T cells, or 1 x 10 6 Or about 1 x 10 6 From 1 × 10 7 Or about 1 x 10 7 The present invention includes administration of total recombinant receptor (e.g., CAR)-expressing T cells or total T cells (each inclusive).

[0152] In some embodiments, the dose of T cells includes CD4 + T cells, CD8 + T cells, or CD4 + And CD8 + Includes T cells.

[0153] In some embodiments, a dose of cells, e.g., recombinant receptor-expressing T cells, is administered to a subject as a single dose, or is administered only once within a period of two weeks, one month, three months, six months, or one year or more.

[0154] In the context of adoptive cell therapy, administration of a certain "dose" includes administration of a given amount or number of cells as a single composition, and / or administration of a single uninterrupted administration, such as administration of a single injection or continuous infusion, and also includes administration of a given amount or number of cells as a split dose or as multiple compositions, provided in multiple separate compositions or infusions over a specified period of time, such as over 3 days or less.Thus, in some situations, a dose is a single or continuous administration of a specified number of cells given or initiated at a single time point.However, in some situations, a dose is administered by multiple injections or infusions over a period of 3 days or less, such as once a day for 3 or 2 days or multiple infusions over 1 day.

[0155] Thus, in some aspects, the dose of cells is administered in a single pharmaceutical composition. In some embodiments, the dose of cells is administered in multiple compositions that collectively comprise the dose of cells.

[0156] In some embodiments, a dose of cells may be administered by administering multiple compositions or solutions, such as a first and a second composition or solution, optionally more compositions or solutions, each of which contains a portion of the dose of cells. In some aspects, multiple compositions, each of which contains different populations and / or subtypes of cells, are administered separately or independently, optionally within a certain period of time. For example, a population or subtype of cells may be administered by administering CD8 + T cells and CD4 + T cells, and / or CD8 + Enriched populations and CD4 + Each enriched population may be, for example, a CD4 + T cells and / or CD8 + In some embodiments, the administration of a dose may include a dose of CD8 T cells. + T cells or a dose of CD4 + Administration of a first composition comprising T cells and a dose of CD4 + T cell dose and CD8 + The administration of a second composition comprising the other of the T cells.

[0157] In some embodiments, administration of a composition or dose, e.g., administration of multiple cell compositions, involves separate administration of the cell compositions. In some aspects, the separate administration occurs simultaneously or sequentially in any order. In certain embodiments, the separate administration involves administration of a dose of CD8 + T cells or a dose of CD4 + A first composition comprising T cells and a dose of CD4 + T cells and CD8 +The administration of the first composition and the second composition comprising the other of the T cells may be performed sequentially in any order. In some embodiments, the dose comprises a first composition and a second composition, and the first composition and the second composition are administered within 48 hours of each other, for example, within 36 hours or less of each other, or within 24 hours or less of each other. In some embodiments, the first composition and the second composition are administered at or about 0 to 12 or about 12 hours apart, at or about 0 to 6 or about 6 hours apart, or at or about 0 to 2 or about 2 hours apart. In some embodiments, the start of administration of the first composition and the start of administration of the second composition are performed at or about 2 hours or less, 1 or about 1 hour or less, or 30 or about 30 minutes or less, 15 or about 15 minutes or less, 10 or about 10 minutes or less, or 5 or about 5 minutes or less apart. In some embodiments, the start and / or end of administration of the first composition and the end and / or start of administration of the second composition occur no more than 2 or about 2 hours, no more than 1 or about 1 hour, or no more than 30 or about 30 minutes apart, no more than 15 or about 15 minutes, no more than 10 or about 10 minutes, or no more than 5 or about 5 minutes apart.

[0158] In some compositions, the first composition, e.g., the first composition of the dose, is a CD4 + In some compositions, the first composition, e.g., the first composition of the dose, comprises CD8 T cells. + In some embodiments, the first composition is administered prior to the second composition. In certain embodiments, the first composition comprises CD8 + T cells are CD4 + Administered before T cells.

[0159] In some embodiments, the dose or composition of cells is determined by a defined or targeted ratio of recombinant receptor (e.g., CAR)-expressing CD4 + CD8 expressing cell-paired recombinant receptor (e.g., CAR) + Cells and / or CD4 + Cells vs. CD8 +In some aspects, the target or desired ratio of different cell populations (e.g., CD4 + :CD8 + Ratio or CAR + CD4 + :CAR + CD8 + Administration of a composition or dose having a ratio (e.g., 1:1) involves administration of a cell composition comprising one of the populations followed by administration of another cell composition comprising the other of the populations, where the administration is at or near a target or desired ratio. In some aspects, administration of a dose of cells or compositions at a defined ratio results in expansion, persistence and / or improved anti-tumor activity of the T cell therapy.

[0160] In some embodiments, the dose or composition of cells comprises a defined or targeted ratio of recombinant CD4 receptors expressing the CD4 receptor. + CD8 expressing cell-paired recombination receptor + Cells and / or CD4 + Cells vs. CD8 + In some aspects, the target or desired ratio of different cell populations (e.g., CD4 + :CD8 + Ratio or CAR + CD4 + :CAR + CD8 + Administration of a composition or dose having a ratio (e.g., 1:1) involves administration of a cell composition comprising one of the populations followed by administration of another cell composition comprising the other of the populations, where the administration is at or near a target or desired ratio. In some aspects, administration of a dose of cells or compositions at a defined ratio results in expansion, persistence and / or improved anti-tumor activity of the T cell therapy.

[0161] In certain embodiments, the number and / or concentration of cells refers to the number of recombinant receptor (e.g., CAR)-expressing cells, or the number of recombinant receptor (e.g., CAR)-expressing T cells, or the CD3+ T cell subset thereof, or the CD4+ and / or CD8+ T cell subset thereof. In some embodiments, the number and / or concentration of cells refers to the number of such cells that are viable cells.

[0162] In some embodiments, a dose of genetically engineered cells is greater than or equal to 2.5×10 7 or about 2.5 x 10 7 CD4+ CAR+ live cells and 2.5×10 7 or about 2.5 x 10 7 Containing separate doses of 5 x 10 CD8+CAR+ live cells 7 or about 5 x 10 7 In some embodiments, the dose of engineered cells is 5×10 7 or about 5 x 10 7 CD4+ CAR+ live cells and 5×10 7 or about 5 x 10 7 Separate doses of 1 x 10 CD8+CAR+ live cells 8 or about 1 x 10 8 In some embodiments, the dose of engineered cells is 0.75×10 8 or about 0.75 x 10 8 CD4+ CAR+ viable cells and 0.75×10 8 or about 0.75 x 10 8 Containing separate doses of 1.5 x 10 live CD8+CAR+ cells 8 or about 1.5 x 10 8 CD3+ CAR+ live cells.

[0163] C. Response, Efficacy, and Survival In some embodiments, the administration effectively treats the subject despite the subject having failed, become resistant and / or resistant to another therapy. In some embodiments, the administration effectively treats the subject despite the subject having become resistant to another therapy. In some embodiments, at least 30% of the subjects treated by the method achieve a complete remission (CR); and / or at least about 75% of the subjects treated by the method achieve an objective response (OR). In some embodiments, at least 35%, 40%, 45%, 50%, 55%, or 60% or more of the subjects treated by the method achieve a CR, and / or at least 50%, 60%, 70%, or 80% or at least about 50%, 60%, 70%, or 80% achieve an objective response (OR). In some embodiments, at least 30% of subjects treated by the method who have failed both a prior BTK inhibitor (e.g., ibrutinib) therapy and venetoclax achieve a complete response (CR); and / or at least about 75% of subjects treated by the method who have failed both a prior BTK inhibitor (e.g., ibrutinib) therapy and venetoclax achieve an objective response (OR). In some embodiments, at least or at least about 35%, 40%, 45%, 50%, 55%, or 60% of subjects treated by the method who have failed both a prior BTK inhibitor (e.g., ibrutinib) therapy and venetoclax achieve a CR, and / or at least or at least about 50%, 60%, 70%, or 80% achieve an objective response (OR). In some embodiments, criteria assessed 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 response), complete remission with incomplete blood count recovery (CRi), stable disease (SD), and / or partial disease (PD).

[0164] In some embodiments, the duration of response before progression is greater than 1 month, greater than 2 months, greater than 3 months, or greater than 6 months or more. In some embodiments, at least 35%, 40%, 45%, 50%, 55%, or 60% or more of subjects treated with the methods provided herein achieve complete remission (CR; also known in some cases as a complete response) at or about 3 months or 6 months or about 6 months after administration of the cell therapy.

[0165] In some aspects, administration according to the provided methods and / or with the provided articles of manufacture or compositions generally reduces or prevents the spread or burden of a disease or condition in a subject. For example, if the disease or condition is a tumor, the method generally reduces tumor size, bulk, metastasis, percent of blast cells in the bone marrow, or molecularly detectable cancer, and / or improves prognosis or survival or other symptoms associated with tumor burden.

[0166] Disease burden may include the total number of disease cells in a subject or in an organ, tissue, or bodily fluid of a subject, e.g., in a tumor or another location, e.g., in an organ or tissue that represents a metastasis. For example, tumor cells may be detected and / or quantified in the blood or bone marrow in the context of certain hematological tumors. Disease burden may, in some embodiments, include tumor mass, number or extent of metastases, and / or percent of blast cells present in the bone marrow.

[0167] In some aspects, the subject has leukemia. The extent of disease burden can be ascertained by assessing residual leukemia in the blood or bone marrow.

[0168] In some aspects, the response rate for a subject, e.g., a subject with CLL, is based on the International Workshop on Chronic Lymphocytic Leukemia (IWCLL) response criteria (Hallek, et al., Blood 2008, Jun 15; 111(12): 5446-5456). In some aspects, the response rate for a subject, e.g., a subject with CLL, is based on the International Workshop on Chronic Lymphocytic Leukemia (IWCLL) response criteria (Hallek et al., Blood 2018 131 (25): 2745-2760). In some aspects, these criteria include: complete remission (CR; sometimes also known as a complete response), which in some aspects requires the absence of peripheral blood clonal lymphocytes by immunophenotyping, the absence of lymphadenopathy, the absence of hepatomegaly or splenomegaly, the absence of constitutional symptoms, and satisfactory blood counts; complete remission with incomplete marrow recovery (CRi), which in some aspects is represented by the above CR but without normal blood counts; partial remission (PR; sometimes also known as a partial response), which in some aspects is represented by a ≥50% decrease in lymphocyte counts, a ≥50% decrease in lymphadenopathy, or a ≥50% decrease in liver or spleen and improvement in peripheral blood counts; progressive disease (PD), which in some aspects is represented by >5x10 9 / L, a ≥50% increase in lymphadenopathy, a ≥50% increase in liver or spleen size, Richter's transformation, or new cytopenias due to CLL; and stable disease, which in some aspects is designated as not meeting the criteria for CR, CRi, PR, or PD.

[0169] In some embodiments, a subject exhibits CR or OR if, within one month of administering the dose of cells, the subject's lymph nodes are less than or about 20 mm in size, less than or about 10 mm in size, or less than or about 10 mm in size.

[0170] In some embodiments, the index clone of CLL is not detectable in the bone marrow of the subject (or in the bone marrow of more than 50%, more than 60%, more than 70%, more than 80%, more than 90% or more of the subjects treated according to the method). In some embodiments, the index clone of CLL is assessed by IgH deep sequencing. In some embodiments, the index clone is not evaluated at, or about, or at least about, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 12 months, 18 months, or 24 months after administration of the cells.

[0171] In some embodiments, the subject shows morphological disease when there are more than or equal to 5% blast cells in the bone marrow, for example, more than or equal to 10% blast cells in the bone marrow, more than or equal to 20% blast cells in the bone marrow, more than or equal to 30% blast cells in the bone marrow, more than or equal to 40% blast cells in the bone marrow, or more than or equal to 50% blast cells in the bone marrow, as detected by light microscopy.In some embodiments, the subject shows complete remission or clinical remission when there are less than 5% blast cells in the bone marrow.

[0172] In some aspects, the subject has leukemia. The extent of disease burden can be ascertained by assessing residual leukemia in the blood or bone marrow.

[0173] In some embodiments, a subject exhibits morphological disease if there are more than 5% or 5% blasts in the bone marrow, e.g., more than 10% or 10% blasts in the bone marrow, more than 20% or 20% blasts in the bone marrow, more than 30% or 30% blasts in the bone marrow, more than 40% or 40% blasts in the bone marrow, or more than 50% or 50% blasts in the bone marrow, e.g., as detected by light microscopy. In some embodiments, a subject exhibits complete or clinical remission if there are less than 5% blasts in the bone marrow.

[0174] In some embodiments, a subject may have a complete remission, but a small percentage of morphologically undetectable (by light microscopy) residual leukemia cells. A subject is said to have minimal residual disease (MRD) if the bone marrow shows less than 5% blast cells and molecularly detectable cancer. In some embodiments, molecularly detectable cancer can be assessed using any of a variety of molecular techniques that allow for sensitive detection of small numbers of cells. In some aspects, such techniques include PCR assays that can ascertain unique Ig / T cell receptor gene rearrangements, or fusion transcripts resulting from chromosomal translocations. In some embodiments, flow cytometry can be used to identify cancer cells based on leukemia-specific immunophenotypes. In some embodiments, molecular detection of cancer can detect one leukemia cell in 100,000 normal cells. In some embodiments, a subject shows molecularly detectable MRD if one leukemia cell is detected in 100,000 cells, for example, by PCR or flow cytometry. In some embodiments, the subject's disease burden is molecularly undetectable, or MRD-, such that, in some cases, leukemia cells are not detectable in the subject using PCR or flow cytometry methods.

[0175] In some embodiments, no indicator clone of leukemia, e.g., CLL, is detected in the bone marrow of the subject (or in the bone marrow of more than 50%, more than 60%, more than 70%, more than 80%, more than 90% or more of the subjects treated according to the method). In some embodiments, indicator clone of leukemia, e.g., CLL, is detected by IGH deep sequencing. In some embodiments, the indicator clone is not detectable at, or about, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 12 months, 18 months, or 24 months after administration of the cells.

[0176] In some aspects, MRD is detected by flow cytometry. Flow cytometry can be used to monitor bone marrow and peripheral blood samples for cancer cells. In certain aspects, flow cytometry is used to detect or monitor the presence of cancer cells in bone marrow. In some aspects, multi-parameter immunological detection by flow cytometry is used to detect cancer cells (see, for example, Coustan-Smith et al., (1998) Lancet 351:550-554). In some aspects, multi-parameter immunological detection by mass cytometry is used to detect cancer cells. In some examples, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45, or 50 parameters can be used to detect cancer cells. The antigens used for detection are selected based on the cancer being detected (Foon and Todd (1986) Blood 68:1-31). In some embodiments, MRD is described as subjects with no evidence of CLL in peripheral blood or bone marrow, i.e., subjects with CR or PR based on residual lymphadenopathy or splenomegaly. In some aspects, MRD is measured via flow cytometry of peripheral blood and IgHV deep sequencing of bone marrow.

[0177] In some cases, bone marrow is collected by bone marrow aspiration or bone marrow biopsy, and lymphocytes are isolated for analysis. To detect epitopes on isolated lymphocytes, such as terminal deoxynucleotidyl transferase (TdT), CD3, CD10, CD11c, CD13, CD14, CD33, CD19, CD20, CD21, CD22, CD23, CD34, CD45, CD56, CD79b, IgM, and / or KORSA3544, monoclonal and / or polyclonal antibodies conjugated to fluorescent dyes (e.g., fluorescein isothiocyanate (FITC), phycoerythrin, peridinin chlorophyll protein, or biotin) can be used. To detect multiple epitopes, the labeled cells can then be detected using flow cytometry, such as multiparameter flow cytometry, or mass cytometry.

[0178] Lymphoid cells can be identified and gated based on light scatter dot plots, followed by a second gate to identify cell populations expressing the immunophenotypic characteristics of interest. Exemplary epitopes are provided in Table 2 below. Other immunological classifications for leukemias and lymphomas include those provided by Foon and Todd Blood (1986) 68(1): 1-31. In some aspects, flow cytometric assessment of MRD may include one or more CLL immunophenotypes (e.g., low levels of forward / side scatter; CD3+; CD4+; CD5+; CD6+; CD8+; CD9+; CD10+; CD11+; CD12+; CD13+; CD14+; CD15+; CD16+; CD17+; CD18+; CD19+; CD20+; CD21+; CD19+; CD22+; CD19+; CD19+; CD22+; CD19+; CD18+; CD21+; CD19+; CD19+; CD22+; CD19+; CD21+; CD19+; CD22+; CD19 ... 陰性 ;CD5 + ;CD14 陰性 ;CD19 + ;CD23 + ;CD45 + ;CD56 陰性 This can be achieved by quantifying live lymphocytes having a .DELTA..times ...

[0179] Table 2. Exemplary immunophenotypic and cytogenetic characteristics TIFF0007652692000002.tif88156+: positive in >90% of cases + / -: positive in >50% of cases - / +: positive in <50% of cases -: positive in <10% of cases Pan-B markers: e.g., CD19, CD20, CD79a sIG: surface immunoglobulin cyIg: cytoplasmic immunoglobulin

[0180] In some aspects, deep sequencing of the immunoglobulin heavy chain (IGH) locus of collected B cells can be used to detect minimal residual disease (MRD). The presence of clones with specific IgG rearrangements can be a marker for detecting the presence of B cell malignancies, such as CLL, and / or the presence of remnants of the malignant cells. In some aspects, cells, such as a population containing B cells or a population suspected to contain B cells, are collected and isolated from blood. In some aspects, cells are collected and isolated from bone marrow, such as bone marrow aspirates or bone marrow biopsies, and / or other biological samples. In some aspects, polymerase chain reaction (PCR) amplification of complementarity determining region 3 (CDR3) is accomplished using primers directed to highly conserved sequences within the V and J regions of the locus, which can be used to identify clonal populations of cells for the purpose of evaluating minimal residual disease. Other methods, such as single cell sequencing approaches, may be used to provide information on the number of cells of a particular lineage and / or to detect clonal populations, including those expressing a particular variable chain, e.g., variable heavy chain or binding site thereof. In some aspects, IGH DNA is amplified using degenerate primers or primers that recognize regions of the variable chain shared between different cell clones, e.g., primers that recognize the consensus V region and the degenerate consensus J region of the IGH sequence. Exemplary sequences of the V region are: An exemplary degenerate consensus sequence for the J region is: The file is TIFF0007652692000004.tif4128.

[0181] In some aspects, the PCR product or sequencing result is specific to the rearranged allele and serves as a clonal marker for MRD detection. After PCR amplification of the CDR3 region, the PCR product can be sequenced to obtain patient-specific oligonucleotides constructed as probes for allele-specific PCR to detect MRD after treatment of B-cell malignancies with CAR-T cell therapy, for example, CD19 CAR-T cell therapy. In examples where PCR products are not generated using consensus primers, V-region family-specific primers against framework region 1 can be used instead.

[0182] In some aspects, the persistence of detectable IGH sequences in tumor cells, such as B-cell malignancies, such as CLL cells, by PCR, such as malignant IGH sequences or detectable IGH sequences corresponding to clonal IGH sequences, after treatment, is associated with increased risk of recurrence.In some aspects, patients who are negative for malignant IGH sequences after treatment (in some aspects, even in the context of other criteria that show progression or partial response, such as persistence of lymphadenopathy, or in some circumstances, other criteria that may be associated with the absence of disease or complete response) may be considered to be more likely to enter CR or durable CR or long-term survival than patients with persistent malignant IGH sequences.In some embodiments, such prognostic and staging determinations are of particular relevance for treatments in which the elimination of malignant cells is observed in a short time after administering therapy, for example, compared to the analysis of other clinical symptoms, such as lymph node size or other staging criteria. For example, in some such aspects, the absence of detectable IGH or minimal residual disease in a sample such as bone marrow may be a favorable readout for response, or the likelihood of response, or its durability, compared to other available staging or prognostic approaches. In some aspects, the results from MRD, such as IGH deep sequencing information, may characterize further intervention or lack thereof. For example, the method and other provided embodiments provide that in some circumstances, a subject who is believed to be negative for malignant IGH may not be further treated with a dose of therapy, as provided in some aspects, or a subject who is believed to be negative for malignant IGH may not be further administered a dose of therapy, as provided in some aspects, or the subject may be administered a lower or reduced dose. Conversely, a subject who shows MRD via IGH deep sequencing may be provided or specified to be further treated, for example, with a similar or higher dose of the initially administered therapy, or with further treatment.In some aspects, the disease or condition persists after administration of the initial dose and / or administration of the initial dose is not sufficient to completely cure the subject's disease or condition.

[0183] In some embodiments, the method reduces the burden of disease or pathology, such as the number of tumor cells, the size of the tumor, the length of patient survival or event-free survival, to a greater extent and / or for a longer period than would be observed using a comparable method. The comparable method uses an alternative administration regimen, such as a regimen that administers one or more alternative therapeutic agents to the subject, and / or a regimen that does not administer a dose of cells and / or a lymphodepleting agent to the subject according to the provided method and / or with the provided article of manufacture or composition. In some embodiments, the burden of disease or pathology in the subject is detected, evaluated, or measured. In some aspects, the disease burden can be detected by detecting the total number of diseased or disease-related cells, such as tumor cells, in the subject, or in the subject's organs, tissues, or bodily fluids, such as blood or serum. In some aspects, the subject's survival rate, survival rate within a certain period of time, degree of survival, presence or duration of event-free survival or symptom-free survival or recurrence-free survival is evaluated. In some embodiments, any symptoms of the disease or pathology are evaluated. In some embodiments, a measure of the burden of a disease or condition is specified.

[0184] In some embodiments, after treatment with the method, the probability of recurrence is reduced compared to other methods, such as methods that do not administer one or more alternative therapeutic agents to the subject and / or methods that do not administer a dose of cells and / or lymphodepleting agents to the subject according to the provided methods and / or with the provided articles of manufacture or compositions.

[0185] In some cases, the pharmacokinetics of administered cells, e.g., adoptively transferred cells, are measured to assess the availability, e.g., bioavailability, of the administered cells. Methods for measuring the pharmacokinetics of adoptively transferred cells may include taking peripheral blood from a subject administered the engineered cells and measuring the number or proportion of engineered cells in the peripheral blood. Approaches for selecting and / or isolating cells may include the use of antibodies specific for chimeric antigen receptors (CARs) (e.g., Brentjens et al., Sci. Transl. Med. 2013 Mar; 5(177): 177ra38); protein L (Zheng et al., J. Transl. Med. 2012 Feb; 10:29); epitope tags such as Strep-Tag sequences directly introduced into specific sites in the CAR (CARs are assessed directly using binding reagents for the Strep-Tag) (Liu et al. (2016) Nature Biotechnology, 34:430; International Patent Application Publication WO2015095895), and monoclonal antibodies that specifically bind to CAR polypeptides (see International Patent Application Publication WO2014190273). In some cases, exogenous marker genes may be used in conjunction with therapies using engineered cells to allow for detection or selection of cells and in some cases to promote cell suicide. In some cases, a truncated epidermal growth factor receptor (EGFRt) can be co-expressed with a transgene of interest (e.g., CAR or TCR) in transduced cells (see, e.g., U.S. Patent No. 8,802,374). EGFRt may contain an epitope recognized by the antibody cetuximab (Erbitux®) or other therapeutic anti-EGFR antibodies or binding molecules, which can be used to identify or select cells engineered with EGFRt constructs and another recombinant receptor, such as a chimeric antigen receptor (CAR), and / or to eliminate or separate cells expressing the receptor.See, e.g., U.S. Patent No. 8,802,374 and Liu et al., Nature Biotech. 2016 April; 34(4): 430-434.

[0186] In some embodiments, CAR in a biological sample obtained from a patient, e.g., blood + The number of T cells can be measured at a period after administration of the cell therapy drug, for example to measure the pharmacokinetics of the cells. In some embodiments, CAR detectable in the blood of the subject or in the majority of subjects so treated by the method. + T cells, optionally CAR + CD8 + T cells and / or CARs + CD4 + The number of T cells is greater than 1 cell / μL, greater than 5 cells / μL, or greater than 10 cells / μL.

[0187] D. Toxicity In some embodiments, the methods provided include, e.g., a method for producing an alternative CAR. + The compositions are designed to include features that result in a lower rate and / or a lower severity of toxicity, toxic outcomes or symptoms, features, factors, or characteristics that promote toxicity, e.g., symptoms or outcomes associated with or indicative of cytokine release syndrome (CRS) or neurotoxicity, as compared to administration of an alternative cellular therapy, such as a T cell composition, and / or an alternative administration of cells, such as an administration of cells not administered at a predetermined ratio.

[0188] In some embodiments, the provided methods do not result in a high rate or high probability of toxicity or toxic outcomes, in other words, lower the rate or probability of toxicity or toxic outcomes, such as neurotoxicity (NT), cytokine release syndrome (CRS), for example, compared to some other cell therapies.In some embodiments, the methods do not result in or 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 3 or more days, and CRP plasma levels of at least 20 mg / dL or at least about 20 mg / dL.In some embodiments, more than 30%, 35%, 40%, 50%, 55%, 60% or more of subjects treated according to the provided methods do not show any grade of CRS or any grade of neurotoxicity. In some embodiments, 50% or less 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) greater than grade 2 and / or neurotoxicity greater than grade 2. In some embodiments, at least 50% of the subjects treated according to the methods (e.g., at least 60%, at least 70%, at least 80%, at least 90% or more of the treated subjects) do not exhibit a severe toxic outcome (e.g., severe CRS or severe neurotoxicity); e.g., do not exhibit grade 3 or higher neurotoxicity and / or do not exhibit severe CRS; or do not do so within a period of time following treatment, e.g., within 1 week, 2 weeks, or 1 month of administration of the cells. In some embodiments, greater than or about 30%, 35%, 40%, 50%, 55%, or 60% or more of subjects who have failed both prior BTK inhibitor (e.g., ibrutinib) therapy and venetoclax treated with the provided methods do not exhibit any grade of CRS or any grade of neurotoxicity.In some embodiments, 50% or less of the treated subjects (e.g., at least 60%, at least 70%, at least 80%, or at least 90% or more of the treated subjects who have failed both previous BTK inhibitor (e.g., ibrutinib) therapy and venetoclax) exhibit cytokine release syndrome (CRS) greater than grade 2 and / or neurotoxicity greater than grade 2. In some embodiments, at least 50% of the treated subjects (e.g., at least 60%, at least 70%, at least 80%, or at least 90% or more of the treated subjects) who have failed both previous BTK inhibitor (e.g., ibrutinib) therapy and venetoclax treated by the method do not exhibit a severe toxic outcome (e.g., severe CRS or severe neurotoxicity), e.g., do not exhibit grade 3 or higher neurotoxicity and / or do not exhibit severe CRS, or do not exhibit it within a certain period of time following treatment, e.g., within 1 week, 2 weeks, or 1 month of administration of the cells. In some embodiments, parameters evaluated to determine a particular toxicity include adverse events (AEs), treatment-emergent adverse events, dose-limiting toxicities (DLTs), CRS, neurological events, and NT.

[0189] The administration of adoptive T cell therapy, for example, treatment with T cells expressing chimeric antigen receptors, can cause toxic effects or outcomes, such as cytokine release syndrome and neurotoxicity. In some instances, such effects or outcomes occur in conjunction with high levels of circulating cytokines, which may underlie the observed toxicity.

[0190] In some aspects, the toxic outcome is, is associated with, or is indicative of cytokine release syndrome (CRS) or severe CRS (sCRS). CRS, for example, sCRS, can occur in some cases after adoptive T cell therapy and after other biological products are administered to a 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.

[0191] Typically, CRS is caused by an exacerbated systemic immune response mediated, for example, by T cells, B cells, NK cells, monocytes, and / or macrophages. Such cells can release a large amount of inflammatory mediators, for example, cytokines and chemokines. Cytokines can trigger acute inflammatory responses and / or induce endothelial organ damage that can cause microvascular leakage, heart failure, or death. Severe, life-threatening CRS can lead to pulmonary infiltration and damage, renal failure, or disseminated intravascular coagulation. Other severe, life-threatening toxicities can include cardiac toxicity, respiratory distress, neurotoxicity, and / or liver failure.

[0192] CRS may be treated with anti-inflammatory therapy, for example, anti-IL-6 therapy, for example, anti-IL-6 antibody, for example, tocilizumab, or antibiotics, or as described. The outcomes, signs, and symptoms of CRS are known and include those described herein. In some embodiments, when a particular regimen or administration results in a certain CRS-related outcome, sign, or symptom, or when a particular regimen or administration does not result in a certain CRS-related outcome, sign, or symptom, a particular outcome, sign, and symptom, and / or its quantity or extent, may be specified.

[0193] In the context of administration of CAR-expressing cells, CRS typically occurs 6-20 days after infusion of the cells expressing the CAR. See Xu et al., Cancer Letters 343 (2014) 172-78. In some cases, CRS occurs less than 6 days or more than 20 days after infusion of the CAR T cells. The incidence and timing of CRS may be related to baseline cytokine levels or tumor burden 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 may be rapidly induced in CRS are IL-1β, IL-6, IL-8, and IL-10.

[0194] Exemplary CRS-related outcomes include fever, rigors, chills, hypotension, dyspnea, acute respiratory distress syndrome (ARDS), encephalopathy, elevated ALT / AST, renal failure, cardiac failure, hypoxia, neurological failure, 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 vascular leak syndrome, liver function decline, and renal failure. In some aspects, CRS is associated with one or more factors, such as increased serum ferritin, d-dimer, aminotransferase, lactate dehydrogenase, and triglycerides, or associated with hypofibrinogenemia or hepatosplenomegaly. Other exemplary signs or symptoms associated with CRS include hemodynamic instability, febrile neutropenia, elevated serum C-reactive protein (CRP), changes in coagulation parameters (e.g., international normalized ratio (INR), prothrombin time (PTI), and / or fibrinogen), changes in cardiac and other organ function, and / or absolute neutrophil count (ANC).

[0195] In some embodiments, CRS-related outcomes include persistent fever, e.g., fever at a specified temperature, e.g., fever of 38° C. or greater, e.g., for more than two days, e.g., more than three days, e.g., for more than four days, or for at least three consecutive days; fever of 38° C. or greater; elevation of cytokines, e.g., a maximum fold change compared to pre-treatment levels of at least two cytokines (e.g., at least two of the group consisting of interferon gamma (IFNγ), GM-CSF, IL-6, IL-10, Flt-3L, fractalkine, and IL-5, and / or tumor necrosis factor alpha (TNFα)), e.g., a maximum fold change of at least 75 or at least about 75, or a maximum fold change of at least one of such cytokines, e.g., a maximum fold change of at least 250 or at least about 250; and / or at least one clinical sign of toxicity, e.g., hypotension (e.g., administration of at least one intravenous vasoactive hypoxia (e.g., less than 90% or less than about 90% plasma oxygen (PO 2 ) level); and / or one or more neurological disorders (including mental status changes, obtundation, and seizures).

[0196] Exemplary CRS-related outcomes include increased or high serum levels of one or more factors, including cytokines and chemokines, and other factors associated with CRS.Furthermore, exemplary outcomes include increased synthesis or secretion of one or more of such factors.Such synthesis or secretion may be by T cells, or cells that interact with T cells, such as innate immune cells or B cells.

[0197] In some embodiments, the CRS-related serum factors or CRS-related outcomes include interferon gamma (IFN-γ), TNF-a, IL-1β, IL-2, IL-6, IL-7, IL-8, IL-10, IL-12, sIL-2Ra, granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage inflammatory protein (MIP)-1, tumor necrosis factor alpha (TNFα), IL-6, and inflammatory cytokines and / or chemokines, including IL-10, IL-1β, IL-8, IL-2, MIP-1, Flt-3L, fractalkine, and / or IL-5. In some embodiments, the factors or outcomes include C-reactive protein (CRP). In addition to being an early and easily measurable risk factor for CRS, CRP is also a marker of cell expansion and proliferation. In some embodiments, subjects who are measured to have a high level of CRP, for example, ≧15 mg / dL, have CRS. In some embodiments, subjects measured to have high levels of CRP do not have CRS, hi some embodiments, measuring CRS includes measuring CRP and another factor indicative of CRS.

[0198] In some embodiments, one or more inflammatory cytokines or chemokines are monitored before, during, or after CAR treatment. In some embodiments, TNF-α or IL-16 are assessed or monitored in a subject, for example, according to the methods described herein.

[0199] To predict which patients are likely at risk for developing sCRS, CRS criteria that appear to correlate with the onset of CRS have been developed (see Davilla et al. Science translational medicine. 2014;6(224):224ra25). Factors include fever, hypoxia, hypotension, neurological changes, and elevated serum levels of inflammatory cytokines, such as a set of seven cytokines (IFNγ, IL-5, IL-6, IL-10, Flt-3L, fractalkine, and GM-CSF) whose treatment-induced elevations can correlate well with pretreatment tumor burden and sCRS symptoms. Other guidelines for the diagnosis and management of CRS are known (see, e.g., Lee et al, Blood. 2014;124(2):188-95). In some embodiments, the criteria reflecting CRS grade are those detailed in Table 3 below.

[0200] Table 3. Exemplary grading criteria for CRS TIFF0007652692000005.tif110164

[0201] In some embodiments, outcomes associated with severe CRS or CRS of grade 3 or higher, e.g., grade 4 or higher, include one or more of the following: persistent fever, e.g., fever above or about 38° C. for two or more, e.g., three or more, e.g., four or more days, or for at least three consecutive days; fever above or about 38° C.; increased cytokines, e.g., increased levels of at least two cytokines (e.g., interferon gamma (IFNγ), GM-CSF, IL-6, IL-10, Flt-3L, Fracta, IL-10 ... and / or at least one clinical sign of toxicity, such as hypotension (e.g., as measured using at least one intravenous vasoactive hypertensive agent); hypoxia (e.g., plasma oxygen (PO) of less than 90% or less than about 90%); 2 ) levels); and / or one or more neurological disorders, including altered mental status, obtundation, and seizures. In some embodiments, severe CRS includes CRS requiring care or nursing in an intensive care unit (ICU).

[0202] 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 CRP serum level of at least 20 mg / dL or at least about 20 mg / dL. In some embodiments, CRS comprises hypotension requiring two or more vasoconstrictors or respiratory failure requiring mechanical ventilation. In some embodiments, the dosage of the vasoconstrictor is increased in the second or subsequent administration.

[0203] 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 tremors.

[0204] Methods for measuring or detecting various outcomes can be specified.

[0205] In some aspects, the toxic outcome is or is associated with neurotoxicity. In some embodiments, symptoms associated with clinical risk of neurotoxicity include confusion, delirium, aphasia, expressive aphasia, obtundation, myoclonus, lethargy, altered mental status, convulsions, seizure-like activity, seizures (optionally confirmed by electroencephalography [EEG]), high levels of beta 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)).

[0206] In some cases, neurological symptoms may be the earliest symptom of sCRS. In some embodiments, neurological symptoms are observed to 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 other symptoms of sCRS have resolved. In some embodiments, the time or extent of resolution of neurological changes is not hastened by treatment with anti-IL-6 and / or steroids.

[0207] In some embodiments, a subject is considered to have "severe neurotoxicity" in response to or associated with administration of a 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 medications) from among: 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, dysesthesias, e.g., distortions of sensory perception causing abnormal and unpleasant sensations, neuralgias, e.g., intense and painful sensations along a nerve or nerve group, and / or paresthesias, e.g., dysfunction of sensory neurons causing abnormal skin sensations of tingling, numbness, pressure, cold, and warmth in the absence of stimulation. In some embodiments, severe neurotoxicity includes grade 3 or higher neurotoxicity, e.g., neurotoxicity as set forth in Table 4.

[0208] Table 4. Exemplary grading criteria for neurotoxicity TIFF0007652692000006.tif66164

[0209] 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, the subject treated according to the method of the present invention may not have detectable symptoms, outcomes, or factors of CRS, such as any of those described, for example, those described in Table 3, for severe CRS or CRS of grade 3 or higher, and / or may have reduced symptoms, outcomes, or factors. In some embodiments, the subject treated according to the method of the present invention may have reduced symptoms of neurotoxicity compared to subjects treated by other methods, such as limb weakness or paralysis; memory, vision, and / or intelligence decline; uncontrollable obsessive and / or compulsive behavior; delusions; headaches; cognitive and behavioral problems, including reduced motor control, cognitive decline, and autonomic nervous system dysfunction; and sexual dysfunction. In some embodiments, a subject treated according to the methods of the invention may have reduced symptoms associated with peripheral motor neuropathy, peripheral sensory neuropathy, dysesthesias, neuralgia, or paresthesia.

[0210] In some embodiments, the method reduces outcomes associated with neurotoxicity, including damage to the nervous system and / or brain, such as neuronal death. In some aspects, the method reduces the levels of factors associated with neurotoxicity, such as beta amyloid (Aβ), glutamate, and oxygen radicals.

[0211] In some embodiments, the toxicity outcome is dose-limiting toxicity (DLT). In some embodiments, the toxicity outcome is dose-limiting toxicity. In some embodiments, the toxicity outcome is the absence of dose-limiting toxicity. In some embodiments, dose-limiting toxicity (DLT) is defined as any toxicity of grade 3 or higher, as assessed by any known or published guideline for evaluating specific toxicity. Such guideline may be, for example, any of those mentioned above, including the National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) version 4.0.

[0212] In some aspects, DLTs are any treatment-emergent Grade 4 or 5 AEs, other than those listed in the exceptions below; any treatment-emergent Grade 3 AEs that do not resolve to Grade ≤ 2 within 7 days, other than those listed in the exceptions below; any treatment-emergent Grade 3 seizures that do not resolve to Grade ≤ 2 within 3 days; and any treatment-emergent Grade 4 or 5 AEs, other than those listed in the exceptions below, that do not resolve to Grade ≤ 2 within 3 days, except for B-cell aplasia (which is an expected risk associated with the administration of engineered cells). could be described as autoimmune toxicity of ≥3; the exceptions listed below are not considered DLTs: any treatment-emergent AE not clearly related to administration of engineered cells (e.g., motor vehicle accident); grade 4 infusion-related toxicity that resolves to grade ≤2 in 8 hours; grade 3 or 4 fever or febrile neutropenia for ≤2 weeks; grade 4 hypertransaminasemia that is considered a symptom of CRS; grade 3 hypertransaminasemia for ≤2 weeks; grade 3 bone pain due to T-cell expansion in the bone marrow compartment for ≤2 weeks; grade 3 or 4 TLS for ≤7 days; grade 3 or 4 hypotension requiring a single vasopressor for port that resolves to grade <3 in ≤72 hours (other no CRS symptoms); Grade 3 or 4 CRS with only hypotension requiring a single vasopressor for support (not requiring intubation) that resolves to Grade <3 in ≤72 hours, or Grade 3 CRS with severity based on Grade 4 transaminasemia; Grade 3 or 4 encephalopathy for ≤7 days that recovers to baseline in ≤14 days from onset of Grade ≥3 event; Grade 3 chills; Grade 3 or 4 lymphopenia; Grade 3 or 4 leukopenia; Grade 3 or 4 asymptomatic electrolyte abnormalities that resolve with supplementation; Grade 3 or 4 thrombocytopenia; Grade 3 or 4 anemia; and Grade 3 or 4 B-cell aplasia and hypogammaglobulinemia.

[0213] In some embodiments, the low rate, risk, or likelihood of developing toxicity, e.g., CRS or neurotoxicity, or severe CRS or neurotoxicity, e.g., Grade 3 or higher CRS or neurotoxicity, observed upon administration of a dose of T cells according to the provided methods and / or with the provided articles of manufacture or compositions allows for administration of the cell therapy on an outpatient basis. + Administration of a dose of the T cells is performed on an outpatient basis or does not require hospitalization of the subject, for example, an overnight stay.

[0214] In some aspects, a cellular therapy, e.g., T cells (e.g., CAR cells), is administered according to the provided methods and / or in conjunction with the provided articles of manufacture or compositions, including to subjects treated on an outpatient basis. + A subject receiving a dose of a cellular antibody (T cells) does not receive any intervention to treat toxicity prior to or with administration of the cell dose unless or until the subject exhibits signs or symptoms of toxicity, e.g., neurotoxicity or CRS. Exemplary agents for treating, delaying, attenuating, or ameliorating toxicity are described in Section II.

[0215] In some embodiments, subjects treated with cell therapy, such as T cells (e.g., CAR +If the subject who receives a dose of T cells) shows fever, the subject is given or instructed to take or administer a therapeutic agent to reduce fever. In some embodiments, the subject's fever is characterized as the subject's body temperature being (or being measured as being) at or above a certain threshold temperature or threshold level. In some aspects, the threshold temperature is associated with at least low fever, at least moderate fever, and / or at least high fever. In some embodiments, the threshold temperature is a certain temperature or range. For example, the threshold temperature may be at or about 38, 39, 40, 41, or 42°C, or at least at or about 38, 39, 40, 41, or 42°C, and / or in the range of 38°C or about 38°C to 39°C or about 39°C, 39°C or about 39°C to 40°C or about 40°C, 40°C or about 40°C to 41°C or about 41°C, or 41°C or about 41°C to 42°C or about 42°C.

[0216] In some embodiments, the treatment designed to reduce fever includes treatment with antipyretic agents.An antipyretic agent can include any agent that reduces fever, such as a compound, composition, or component, for example, any one of a number of agents known to have antipyretic effects, such as NSAIDs (such as ibuprofen, naproxen, ketoprofen, and nimesulide), salicylates, such as aspirin, choline salicylate, magnesium salicylate, and sodium salicylate, paracetamol, acetaminophen, metamizole, nabumetone, phenaxone, antipyrine, antipyretic drugs, etc.In some embodiments, the antipyretic agent is acetaminophen.In some embodiments, acetaminophen can be administered orally or intravenously at a dose of 12.5mg / kg, with a maximum dose every 4 hours.In some embodiments, the antipyretic agent is ibuprofen or aspirin, or includes ibuprofen or aspirin.

[0217] In some embodiments, if the fever is persistent fever, the subject is administered an alternative therapeutic agent for treating toxicity, such as any of those described in section II below.For subjects treated as outpatients, if the subject has and / or is determined to have persistent fever, the subject is instructed to return to hospital.In some embodiments, the subject is determined or considered to have and / or have persistent fever when: after a designated treatment, such as a treatment designed to reduce fever, such as treatment with antipyretic, e.g., NSAID or salicylates, e.g., ibuprofen, acetaminophen, or aspirin, the subject shows a fever of a reasonable threshold temperature or higher, and the subject's fever or body temperature does not decrease or does not decrease by a designated amount or more than a designated amount (e.g., does not decrease by more than 1°C, generally does not fluctuate by about 0.5°C, 0.4°C, 0.3°C, or 0.2°C, or does not fluctuate 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 the subject exhibits, or is determined to exhibit, a fever of at least or at least about 38 or 39° C., and the 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., or by more than 0.5° C., 0.4° C., 0.3° C., or 0.2° C., or by less than 1%, 2%, 3%, 4%, or 5%, or by about 1%, 2%, 3%, 4%, or 5%, over 6, 8, 12, or 24 hours, even after treatment with an antipyretic agent such as acetaminophen. In some embodiments, the dosage of the antipyretic agent is one that is normally effective in reducing a fever or a particular type of fever, such as a fever associated with a bacterial or viral infection, such as a localized or systemic infection.

[0218] In some embodiments, a subject is determined or considered to have and / or have a persistent fever if the subject exhibits a fever at or above a reasonable threshold temperature, and the subject's fever or body temperature does not fluctuate by more than about 1°C or about 1°C, generally by about 0.5°C, 0.4°C, 0.3°C, or 0.2°C, or by about 0.5°C, 0.4°C, 0.3°C, or 0.2°C. Typically, such fluctuations of more than a certain amount or no fluctuations of a certain amount are measured over a period of time (e.g., over a period of 24 hours, 12 hours, 8 hours, 6 hours, 3 hours, or 1 hour, and may be measured from the first sign of fever or from when the body temperature first exceeds a specified threshold). For example, in some embodiments, a subject is considered or determined to exhibit a persistent fever if the subject exhibits a fever of at least or at least about 38 or 39° C., or at least or at least about 38 or 39° C., and the body temperature does not fluctuate by more than or about 0.5° C., 0.4° C., 0.3° C., or 0.2° C. over 6 hours, over 8 hours, over 12 hours, or over 24 hours.

[0219] In some embodiments, the fever is a persistent fever; in some aspects, the subject is diagnosed with a persistent fever at the time of such diagnosis, e.g., from such diagnosis or from an initial therapy that may induce toxicity, e.g., a cell therapy, e.g., a CAR- + Treatment occurs within 1, 2, 3, 4, 5, 6, or less hours from the first such determination following administration of T cells, such as T cells.

[0220] In some embodiments, one or more interventions or agents for treating toxicity, e.g., toxicity-targeted therapeutics, are administered at or shortly after a subject is determined or confirmed (e.g., initially determined or confirmed) to have persistent fever, e.g., as measured according to any of the preceding embodiments. In some embodiments, one or more toxicity-targeted therapeutics are administered within a period of such confirmation or determination, e.g., within 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, or 8 hours thereafter.

[0221] E. Biomarkers Among the methods provided are methods of treatment involving assessing the risk of developing toxicity associated with cell therapy in a subject, involving assessing or detecting a biomarker (e.g., analyte) or parameter associated with toxicity, e.g., neurotoxicity, e.g., severe neurotoxicity, and / or CRS, e.g., severe CRS. In some aspects, the subject is administered a therapy, e.g., engineered T cell therapy, based on the assessment. In some embodiments, the biomarkers include TNF-α (TNFα) and interleukin-16 (IL-16).

[0222] In some embodiments, the method involves assessing or detecting the presence or absence of a biomarker (e.g., TNFα or IL-16) and / or a parameter (e.g., concentration, amount, level, or activity) associated with the biomarker (e.g., TNFα or IL-16). In some cases, the method may include comparing one or more parameters to a certain reference value, e.g., a threshold level (also referred to herein as a "threshold"), e.g., one associated with a risk of developing toxicity, such as CRS or NT, and / or severe toxicity, e.g., severe CRS or NT. In some embodiments, the method also involves selecting a subject for treatment with a cell therapy based on the assessment of the presence or absence of a biomarker and / or the comparison of the biomarker to a reference value or threshold level of the biomarker. In some embodiments, the method also involves administering an agent or therapy that can treat, prevent, delay, and / or attenuate the onset of toxicity, e.g., based on the assessment of the presence or absence of a biomarker and / or the comparison of the biomarker to a reference value or threshold level of the biomarker.

[0223] In some embodiments, the method involves assessing the risk of developing toxicity before and / or after administering cell therapy.In some embodiments, the method involves assessing the level, amount, or concentration of a biomarker (e.g., TNFα or IL-16) in a biological sample, where the biological sample is from a subject who is a candidate for cell therapy treatment, and the cell therapy optionally includes a dose or composition of genetically engineered cells expressing a recombinant receptor; and the biological sample is obtained from the subject before administering cell therapy, and / or the biological sample does not include recombinant receptor and / or the engineered cells.In some aspects, the method involves comparing the level, amount, or concentration of an analyte in the sample with a threshold level individually, thereby determining the risk of developing toxicity after administering cell therapy.In some aspects, the comparison can be used to determine the risk of developing toxicity after administering cell therapy.

[0224] In some aspects, the method comprises: assaying a biological sample for a level, amount, or concentration of TNF-α, wherein the biological sample is optionally derived from a subject who is a candidate for treatment with a cell therapy, and the cell therapy comprises a dose of engineered cells, including T cells expressing a CAR for treating a disease or condition, and wherein the biological sample was obtained from the subject prior to administering the cell therapy and / or the biological sample does not comprise a CAR and / or the engineered cells; and comparing the level, amount, or concentration of TNF-α to a threshold level, (1) identifying the subject as at risk for developing grade 3 or greater neurotoxicity following administration of the cell therapy if the level, amount, or concentration of TNF-α is at or above the threshold level; and (2) identifying the subject as not at risk for developing grade 3 or greater neurotoxicity following administration of the cell therapy if the level, amount, or concentration of TNF-α is below the threshold level. This is accompanied by:

[0225] In some embodiments, if the subject is identified as at risk for developing grade 3 or higher neurotoxicity, the method further involves: (i) administering to the subject a cell therapy, optionally at a reduced dose, where optionally (a) the method further comprises administering to the subject an agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset or risk of onset of neurotoxicity, and / or (b) administration of the cell therapy to the subject is performed or designated to be performed in an in-patient setting and / or with one or more days of hospitalization; or (ii) administering to the subject an alternative treatment other than the cell therapy to treat the disease or condition.

[0226] In some embodiments, if a subject is identified as not at risk for developing Grade 3 or higher neurotoxicity following administration of a cell therapy: (i) optionally at or after the subject exhibits persistent fever or fever that falls or does not fall or does not fall by more than 1° C. following treatment with an antipyretic, the subject is not administered an agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset of, or the risk of onset of, toxicity, unless or until the subject exhibits signs or symptoms of toxicity; and / or (ii) optionally unless or until the subject exhibits persistent fever or fever that falls or does not fall or does not fall by more than 1° C. following treatment with an antipyretic, the administration and all follow-up is performed on an outpatient basis and / or without admitting the subject to a hospital and / or without an overnight stay in a hospital and / or without the need for hospitalization or overnight stay in a hospital.

[0227] In some embodiments, the assay method involves: (a) contacting the biological sample with one or more reagents capable of detecting TNF-α or specific for TNF-α, optionally wherein the one or more reagents include an antibody that specifically recognizes TNF-α; and (b) detecting the presence or absence of a complex comprising the one or more reagents and TNF-α.

[0228] In some embodiments, the methods involve administering to a subject a cell therapy for treating a disease or condition, the cell therapy comprising a dose of engineered cells comprising T cells expressing a CAR, wherein (1) if the subject has a level, amount, or concentration of TNF-α in a biological sample from the subject that is at or above a threshold level, the subject is identified as at risk for developing Grade 3 or greater neurotoxicity following administration of the cell therapy: (i) administering the cell therapy at a reduced dose to the subject; (ii) further administering to the subject an agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset or risk of onset of toxicity; and / or (iii) administering to the subject the cell therapy is performed or designated to be performed in an inpatient setting and / or with one or more days of hospitalization; or (2) if the subject is selected or identified as having a level, amount, or concentration of TNF-α in a biological sample from the subject that is below a threshold level, the subject is identified as at risk for developing Grade 3 or greater neurotoxicity following administration of the cell therapy. identified as not at risk for developing neurotoxicity, (i) optionally at or after the subject exhibits persistent fever or a fever that drops or does not drop or does not drop by more than 1° C. following treatment with an antipyretic, unless or until the subject shows signs or symptoms of toxicity, said administration and all follow-up is performed on an outpatient basis and / or without the subject being admitted to a hospital and / or without an overnight stay in a hospital and / or without the need for hospitalization or overnight stay in a hospital, unless or until the subject shows persistent fever or a fever that drops or does not drop by more than 1° C. following treatment with an antipyretic, said administration and all follow-up is performed on an outpatient basis and / or without the subject being admitted to a hospital and / or without an overnight stay in a hospital, and / or the subject is a candidate for treatment with a cell therapy, and said biological sample and / or said biological sample obtained from the subject prior to administration of cell therapy does not contain a CAR and / or said engineered cells.

[0229] In some embodiments, if the subject is identified as at risk for developing grade 3 or greater neurotoxicity following administration of the cell therapy, the method involves administering an agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset or risk of onset of toxicity, where the agent is administered to the subject simultaneously with the cell therapy or within 3 days of administration of the cell therapy to the subject.

[0230] In some embodiments, the method includes: (a) assaying a biological sample from a subject receiving a cell therapy comprising a dose of engineered cells comprising T cells expressing a CAR to treat a disease or condition for a level, amount, or concentration of IL-16, wherein the biological sample is obtained from the subject within 1, 2, or 3 days after initiation of administration of the cell therapy; and (b) comparing the level, amount, or concentration of IL-16 to a threshold level, wherein (1) if the level, amount, or concentration of IL-16 is at or above the threshold level, then the subject is identified as at risk for developing grade 3 or higher neurotoxicity; and (2) if the level, amount, or concentration of IL-16 is below the threshold level, then the subject is identified as not at risk for developing grade 3 or higher neurotoxicity.

[0231] In some embodiments, if the subject is identified as at risk of developing grade 3 or higher neurotoxicity, the method involves administering an agent or other treatment that can treat, prevent, delay, reduce, or attenuate the onset of toxicity or the risk of developing the toxicity. In some embodiments, the assaying involves: (a) contacting the biological sample with one or more reagents capable of detecting IL-16 or specific for IL-16, optionally comprising an antibody that specifically recognizes IL-16; and (b) detecting the presence or absence of a complex comprising the one or more reagents and IL-16. In some embodiments, the method involves administering a cell therapy to the subject prior to the assaying.

[0232] In some embodiments, the method involves administering to a subject identified as at risk of developing grade 3 or higher neurotoxicity an agent or other treatment capable of treating, preventing, delaying, reducing, or attenuating the onset or risk of onset of toxicity, where the subject has previously received cell therapy to treat a disease or condition, and the subject is selected or identified as at risk of developing grade 3 or higher neurotoxicity at or just prior to administering the agent if the level or amount or concentration of IL-16 in a biological sample obtained from the subject within 1, 2, or 3 days of the start of administration of the cell therapy is above a threshold level. In some embodiments, administering the agent occurs when the subject exhibits a persistent fever or a fever that is lowered by more than 1° C. or is not lowered or does not lower after treatment with an antipyretic. In some embodiments, administration of cell therapy to the subject is performed on an outpatient basis, and if the level, amount, or concentration of IL-16 is above a threshold level, the method includes hospitalizing the patient in a hospital for one or more days.

[0233] In some embodiments, the biological sample is or is obtained from a blood, plasma, or serum sample. In some embodiments, the assaying step comprises an immunoassay. In some embodiments, a cellular therapy for use in a method of treatment is provided, the method comprising: assaying a biological sample for a level, amount, or concentration of TNF-α, wherein the biological sample is optionally derived from a subject who is a candidate for treatment with a cell therapy, and the cell therapy comprises a dose of engineered cells, including T cells expressing a CAR for treating a disease or condition, and wherein the biological sample was obtained from the subject prior to administering the cell therapy and / or the biological sample does not comprise a CAR and / or the engineered cells; comparing the level, amount, or concentration of TNF-α to a threshold level, where (1) if the level, amount, or concentration of TNF-α is at or above the threshold level, then identifying the subject as at risk for developing Grade 3 or higher neurotoxicity following administration of the cell therapy; and (2) if the level, amount, or concentration of TNF-α is below the threshold level, then identifying the subject as not at risk for developing Grade 3 or higher neurotoxicity following administration of the cell therapy; and administering a cellular therapy, optionally at a reduced dose, to the subject, optionally comprising: (a) the method further comprises administering to the subject an agent or other treatment capable of treating, preventing, delaying, reducing or attenuating the onset or risk of onset of neurotoxicity; and / or (b) administration of the cellular therapy to the subject is performed or designated to be performed in an inpatient setting and / or with a one or more day hospital stay; step.

[0234] In some of the optional embodiments, an agent or other treatment capable of treating, preventing, delaying, reducing, or attenuating the onset or risk of onset of neurotoxicity is provided for use in a method of reducing neurotoxicity following administration of cell therapy, the method comprising: assaying a biological sample for a level, amount, or concentration of TNF-α, wherein the biological sample is optionally derived from a subject who is a candidate for treatment with a cell therapy, and the cell therapy comprises a dose of engineered cells, including T cells expressing a CAR for treating a disease or condition, and wherein the biological sample was obtained from the subject prior to administering the cell therapy and / or the biological sample does not comprise a CAR and / or the engineered cells; comparing the level, amount, or concentration of TNF-α to a threshold level, where (1) if the level, amount, or concentration of TNF-α is at or above the threshold level, then identifying the subject as at risk for developing Grade 3 or higher neurotoxicity following administration of the cell therapy; and (2) if the level, amount, or concentration of TNF-α is below the threshold level, then identifying the subject as not at risk for developing Grade 3 or higher neurotoxicity following administration of the cell therapy; and administering agents or other treatments and cell therapies to the subject, optionally at reduced doses. and / or (b) administration of the cellular therapy to the subject is performed or designated to be performed in an inpatient setting and / or with a one or more day stay in a hospital.

[0235] In some aspects, provided herein is a cell therapy for use in a method of treatment, the method being based on a step of administering the cell therapy to a subject, optionally at a reduced dose, where optionally (a) the method further comprises administering to the subject an agent or other treatment capable of treating, preventing, delaying, reducing, or attenuating the onset or risk of onset of neurotoxicity, and / or (b) the administration of the cell therapy to the subject is or is designated to be administered in an inpatient setting and / or with one or more days of hospitalization, where the patient is a candidate for treatment with a cell therapy comprising a dose of engineered cells comprising T cells expressing a CAR for treating a disease or condition, the method comprising assaying a biological sample from a subject who is a candidate for treatment with a cell therapy comprising a dose of engineered cells comprising T cells expressing a CAR for treating a disease or condition. and wherein the subject has been identified as at risk for developing grade 3 or greater neurotoxicity following administration of the cell therapy, and the biological sample is obtained from the subject prior to administration of the cell therapy and / or the biological sample does not contain a CAR and / or the engineered cells for a level, amount, or concentration of TNF-α, and comparing the level, amount, or concentration of TNF-α to a threshold level, where (1) if the level, amount, or concentration of TNF-α is at or above the threshold level, the subject is identified as at risk for developing grade 3 or greater neurotoxicity following administration of the cell therapy; and (2) if the level, amount, or concentration of TNF-α is below the threshold level, the subject is identified as not at risk for developing grade 3 or greater neurotoxicity following administration of the cell therapy.

[0236] In some aspects, provided herein are agents or other treatments that can treat, prevent, delay, reduce, or attenuate the onset or risk of onset of neurotoxicity for use in methods of reducing neurotoxicity following administration of cell therapy, the methods comprising: administering to the subject a cell therapy, optionally at a reduced dose, and an agent or other treatment, optionally wherein administration of the cell therapy and agent or other treatment to the subject is or is designated to be administered in an inpatient setting and / or with one or more days of hospitalization, the patient having been identified as at risk for developing grade 3 or greater neurotoxicity following administration of the cell therapy based on a method comprising assaying a biological sample from the subject who is a candidate for treatment with a cell therapy comprising a dose of engineered cells comprising T cells expressing a CAR to treat a disease or condition, the biological sample being obtained from the subject prior to administration of the cell therapy and / or the biological sample not comprising a CAR and / or the engineered cells for a level, amount, or concentration of TNF-α; and Comparing the level, amount, or concentration of TNF-α to a threshold level, (1) identifying the subject as at risk for developing grade 3 or higher neurotoxicity after administration of the cell therapy if the level, amount, or concentration of TNF-α is at or above the threshold level; and (2) identifying the subject as not at risk for developing grade 3 or higher neurotoxicity after administration of the cell therapy if the level, amount, or concentration of TNF-α is below the threshold level.

[0237] II. Interventions or Agents for Treating or Ameliorating Toxic Symptoms In some embodiments, the methods and articles of manufacture provided may be used in conjunction with, or may involve, one or more agents or treatments for treating, preventing, delaying, or attenuating the onset of toxicity. In some examples, an agent or other treatment that can treat, prevent, delay, or attenuate the onset of toxicity is administered prior to and / or concurrently with administration of a therapeutic cell composition comprising genetically engineered cells.

[0238] In some embodiments, the agent (e.g., a toxicity targeting agent) or treatment that can treat, prevent, delay, or attenuate the onset of toxicity is a steroid, an antagonist or inhibitor of a cytokine receptor (e.g., IL-6 receptor, CD122 receptor (IL-2R beta receptor), or CCR2), or an inhibitor of a cytokine (e.g., IL-6, MCP-1, IL-10, IFN-γ, IL-8, or IL-18). In some embodiments, the agent is an agonist of a cytokine receptor and / or cytokine, e.g., TGF-β. In some embodiments, the agent, e.g., an agonist, antagonist, or inhibitor, is an antibody or antigen-binding fragment, a small molecule, a protein or peptide, or a nucleic acid.

[0239] In some embodiments, a fluid bolus may be used as an intervention, for example to treat the hypotension associated with CRS. In some embodiments, the target hematocrit level is >24%. In some embodiments, the intervention includes the use of absorbent resin techniques to filter blood or plasma. In some cases, the intervention includes dialysis, plasmapheresis, or similar techniques. In some embodiments, vasopressors or acetaminophen may be used.

[0240] In some embodiments, the agents can be administered sequentially, intermittently, or simultaneously with the therapy (e.g., cells for adoptive cell therapy), or in the same composition. For example, the agents can be administered before, during, simultaneously with, or after administration of immunotherapy and / or cell therapy.

[0241] In some embodiments, the agent is administered at the time described herein, according to the method provided, and / or in the article of manufacture or composition provided.In some embodiments, the toxic targeting agent is administered, for example, less than 3, 4, 5, 6, 7, 8, 9, or 10 days after the start of immunotherapy and / or cell therapy, or within a range of 3, 4, 5, 6, 7, 8, 9, or 10 days or less.In some embodiments, the toxic targeting agent is administered within 1 or about 1 day, within 2 or about 2 days, or within 3 or about 3 days after the start of administration of immunotherapy and / or cell therapy.

[0242] In some embodiments, the agent (e.g., toxic targeting agent) is administered to the subject after the start of administering immunotherapy and / or cell therapy, when the subject does not show grade 2 or higher CRS or grade 2 or higher neurotoxicity.In some aspects, the toxic targeting agent is administered to the subject after the start of administering immunotherapy and / or cell therapy, when the subject does not show severe CRS or severe neurotoxicity.Therefore, between the start of administering immunotherapy and / or cell therapy and the start of administering toxic targeting agent, the subject is not showing grade 2 or higher CRS, e.g., severe CRS, and / or not showing grade 2 or higher neurotoxicity, e.g., severe neurotoxicity.

[0243] Non-limiting examples of interventions for the treatment or amelioration of toxicity, such as severe CRS (sCRS), or severe neurotoxicity, are shown in Table 5. In some embodiments, the intervention includes tocilizumab or other toxicity targeting agents as described, and can be at the time the subject experiences a persistent or persistent fever above or about 38° C., or above or about 39° C. In some embodiments, the fever has persisted in the subject for more than 10 hours, more than 12 hours, more than 16 hours, or more than 24 hours prior to the intervention.

[0244] Table 5. Exemplary Interventions TIFF0007652692000007.tif114165

[0245] In some cases, the agent or therapy or intervention (e.g., toxic targeting agent) is administered alone or as part of a composition or formulation (e.g., pharmaceutical composition or formulation described herein).Accordingly, the agent alone or as part of a pharmaceutical composition can be administered intravenously or orally, or by any other acceptable known administration route or as described herein.

[0246] In some embodiments, the dosage or frequency of administration of the agent in the dosing regimen is less than the dosage or frequency of administration of the agent in a method in which a subject is treated with the agent after developing or being diagnosed with grade 2 or higher CRS or neurotoxicity, e.g., severe, e.g., grade 3 or higher CRS, or severe, e.g., grade 3 or higher neurotoxicity (e.g., after physical signs or symptoms of grade 3 or higher CRS or neurotoxicity appear). In some embodiments, the dosage or frequency of administration of the agent in the dosing regimen is less than the dosage or frequency of administration of the agent in a method in which a subject is treated for CRS or neurotoxicity for more than 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks or more after administration of immunotherapy and / or cell therapy. In some embodiments, the dosage is reduced by more than or about 1.2-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold or more. In some embodiments, the dosage is reduced by more than or about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more. In some embodiments, the dosage is reduced less frequently, e.g., administered fewer times per day or for fewer days.

[0247] A. Steroids In some embodiments, the agent (e.g., a toxicity targeting agent) that treats and / or prevents, delays, or attenuates the development or risk of toxicity to immunotherapy and / or cell therapy is a steroid, such as a corticosteroid. Corticosteroids typically include glucocorticoids and mineralocorticoids.

[0248] Any corticosteroid (e.g., glucocorticoid) may be used in the methods provided herein. In some embodiments, glucocorticoids include synthetic and non-synthetic glucocorticoids. Exemplary glucocorticoids include, but are not limited to, alclometasone, algestone, beclomethasone (e.g., beclomethasone dipropionate), betamethasone (e.g., betamethasone 17-valerate, betamethasone sodium acetate, betamethasone sodium phosphate, betamethasone valerate), budesonide, clobetasol (e.g., clobetasol propionate), clobetasone, clocortolone (e.g., clocortolone pivalate), cloprednol, corticosterone. , cortisone and hydrocortisone (e.g., hydrocortisone acetate), cortivazol, deflazacort, desonide, desoximetasone, dexamethasone (e.g., dexamethasone 21-phosphate, dexamethasone acetate, dexamethasone sodium phosphate), diflorasone (e.g., diflorasone diacetate), diflucortolone, difluprednate, enoxolone, fluazacort, flucloronide, fludrocortisone (e.g., for example, fludrocortisone acetate), flumethasone (e.g., flumethasone pivalate), flunisolide, fluocinolone (e.g., fluocinolone acetonide), fluocinonide, fluocortin, fluocortolone, fluorometholone (e.g., fluorometholone acetate), fluperolone (e.g., fluperolone acetate), fluprednidene, fluprednisolone, flurandrenolide, fluticasone (e.g., fluticasone propionate), formocortal, halcinonide, halobetasol , halometasone, halopredone, hydrocortamate, hydrocortisone (e.g., hydrocortisone 21-butyrate, hydrocortisone aceponate, hydrocortisone acetate, hydrocortisone buteprate, hydrocortisone butyrate, hydrocortisone cypionate, hydrocortisone hemisuccinate, hydrocortisone probutate, hydrocortisone sodium phosphate, hydrocortisone sodium succinate, hydrocortisone valerate), loteprednol etabonate,mazipredone, medrysone, meprednisone, methylprednisolone (methylprednisolone aceponate, methylprednisolone acetate, methylprednisolone hemisuccinate, methylprednisolone sodium succinate), mometasone (e.g., mometasone furoate), paramethasone (e.g., paramethasone acetate), prednicarb, prednisolone (e.g., prednisolone 25-diethylaminoacetate, prednisolone sodium phosphate, prednisolone 21-hemisuccinate, prednisolone acetate; prednisolone farnesylate, hemisuccinate Prednisolone succinate, prednisolone-21 (beta-D-glucuronide), prednisolone metasulfobenzoate, prednisolone stearate, prednisolone tebutate, prednisolone tetrahydrophthalate), prednisone, prednival, prednylidene, rimexolone, tixocortol, triamcinolone (e.g., triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide, triamcinolone acetonide 21-palmitate, triamcinolone diacetate). Such glucocorticoids and their salts are described in detail, for example, in Remington's Pharmaceutical Sciences, A. Osol, ed., Mack Pub. Co., Easton, Pa. (16th ed. 1980).

[0249] In some instances, the glucocorticoid is selected from among cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, and prednisone. In certain instances, the glucocorticoid is dexamethasone.

[0250] In some embodiments, the agent is a corticosteroid and is administered in a therapeutically effective amount to treat, ameliorate, or reduce one or more symptoms of toxicity (e.g., CRS or neurotoxicity) to immunotherapy and / or cell therapy. In some embodiments, indicators of improvement or successful treatment include a determination that a toxicity grading scale (e.g., a CRS or neurotoxicity grading scale) does not score (e.g., a score below 3), or a change in grading or severity on a grading scale described herein, such as a change from a score of 4 to a score of 3, or a change from a score of 4 to a score of 2, 1, or 0.

[0251] In some aspects, the corticosteroid is provided in a therapeutically effective dose. The therapeutically effective concentration can be empirically determined by testing in known in vitro or in vivo (e.g., animal model) systems. For example, the amount of a selected corticosteroid administered to ameliorate symptoms or adverse effects of toxicity (e.g., CRS or neurotoxicity) to immunotherapy and / or cell therapy can be determined by standard clinical procedures. In addition, animal models can be used to help identify optimal dosage ranges. The exact dosage that can be empirically determined can depend on the specific therapeutic preparation, regimen, and administration schedule, route of administration, and severity of the disease.

[0252] The corticosteroid can be administered in any amount effective to ameliorate one or more symptoms associated with the toxicity (eg, CRS or neurotoxicity). Corticosteroids (e.g., glucocorticoids) may be administered in amounts of, for example, at or about, 0.1 to 100 mg, 0.1-80 mg, 0.1-60 mg, 0.1-40 mg, 0.1-30 mg, 0.1-20 mg, 0.1-15 mg, 0.1-10 mg, 0.1-5 mg, 0.2-40 mg, 0.2-30 mg, 0.2-20 mg, 0.2-15 mg, 0.2-10 mg, 0.2-5 mg, 0.4-40 mg, 0.4-30 mg, 0.4-20 mg, 0.4-15 mg, 0.4-10 mg, 0.4-5 mg, 0.4-4 mg, 1-20 mg, 1-15 mg, or 1-10 mg per dose for a 70 kg adult human subject. Typically, a corticosteroid (e.g., a glucocorticoid) will be administered at or about 0.4 to 20 mg per dose for an average adult human subject, e.g., 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.75 mg, 0.8 mg, 0.9 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg or 20 mg per dose. It is administered in amounts of 18 mg, 19 mg or 20 mg.

[0253] In some embodiments, the corticosteroid is administered at a dose of 0.001 mg / kg, 0.002 mg / kg, 0.003 mg / kg, 0.004 mg / kg, 0.005 mg / kg, 0.006 mg / kg, 0.007 mg / kg, 0.008 mg / kg, 0.009 mg / kg, 0.01 mg / kg, 0.015 mg / kg, 0.02 mg / kg, 0.025 mg / kg, 0.03 mg / kg, 0.035 mg / kg, 0.04 mg / kg, 0.045 mg / kg, 0.05 mg / kg, 0.055 mg / kg, 0.06 mg / kg, 0.065 mg / kg, 0.07 mg / kg, 0.075 mg / kg, 0.08 mg / kg, 0.085 mg / kg, 0.095 ...95 mg / kg, 0.01 mg / kg, 0.015 mg / kg, 0.02 mg / kg, 0.025 mg / kg, 0.03 mg / kg, 0.035 mg / kg, 0.04 mg / kg, 0.045 mg / kg, 0.09 mg / kg, 0.095 mg / kg, 0.1 mg / kg, 0.15 mg / kg, 0.2 mg / kg, 0.25 mg / kg, 0.30 mg / kg, 0.35 mg / kg, 0.40 mg / kg, 0.45 mg / kg, 0.50 mg / kg, 0.55 mg / kg, 0.60 mg / kg, 0.65 mg / kg, 0.70 mg / kg, 0.75 mg / kg, 0.80 mg / kg, 0.85 mg / kg, 0.90 mg / kg, 0.95 mg / kg, 1 mg / kg, 1.05 mg / kg, 1.1 mg / kg, 1.15 mg / kg, 1.20 mg / kg, 1.25 mg / kg, 1.3 mg / kg, 1.35 mg / kg or 1.4 mg / kg or about 0.001 mg / kg, 0.002 mg / kg, 0.003 mg / kg, 0.004 mg / kg, 0.005 mg / kg, 0.006 mg / kg, 0.007 mg / kg, 0.008 mg / kg, 0.009 mg / kg, 0.009 mg / kg, 0.1 mg / kg, 0.15 mg / kg, 0.2 mg / kg, 0.25 mg / kg, 0.30 mg / kg, 0.35 mg / kg, 0.40 mg / kg, 0.45 mg / kg, 0.50 mg / kg, 0.55 mg / kg, 0.60 mg / kg, 0.65 mg / kg, 0.70 mg / kg, 0.75 mg / kg, 0.80 mg / kg, 0.85 mg / kg, 0.90 mg / kg, mg / kg, 0.003 mg / kg, 0.004 mg / kg, 0.005 mg / kg, 0.006 mg / kg, 0.007 mg / kg, 0.008 mg / kg, 0.009 mg / kg, 0.01 mg / kg, 0.015 mg / kg, 0.02 mg / kg, 0.025 mg / kg, 0.03 mg / kg, 0.035 mg / kg, 0.04 mg / kg, 0.045 mg / kg, 0.05 mg / kg, 0.055 mg / kg, 0.06 mg / kg, 0.065 mg / kg, 0.07 mg / kg, 0.075 mg / kg, 0.08 mg / kg, 0.085 mg / kg, 0.09 mg / kg, 0.095 mg / kg, 0.It may be administered at a dosage of 1 mg / kg, 0.15 mg / kg, 0.2 mg / kg, 0.25 mg / kg, 0.30 mg / kg, 0.35 mg / kg, 0.40 mg / kg, 0.45 mg / kg, 0.50 mg / kg, 0.55 mg / kg, 0.60 mg / kg, 0.65 mg / kg, 0.70 mg / kg, 0.75 mg / kg, 0.80 mg / kg, 0.85 mg / kg, 0.90 mg / kg, 0.95 mg / kg, 1 mg / kg, 1.05 mg / kg, 1.1 mg / kg, 1.15 mg / kg, 1.20 mg / kg, 1.25 mg / kg, 1.3 mg / kg, 1.35 mg / kg or 1.4 mg / kg.

[0254] The corticosteroid or glucocorticoid (e.g., dexamethasone) may be administered orally (tablet, liquid, or concentrated liquid), PO, intravenously (IV), intramuscularly, or by any other route known or described herein (e.g., with respect to pharmaceutical formulations). In some aspects, the corticosteroid is administered as a bolus, and in other aspects over a period of time.

[0255] In some aspects, glucocorticoids can be administered for a period of more than one day, for example, for two days, three days, or four days, or longer.In some embodiments, corticosteroids can be administered once a day, twice a day, or three times a day, or more.For example, corticosteroids (e.g., dexamethasone) can be administered in some instances at 10 mg (or equivalent) twice a day by IV for three days.

[0256] In some embodiments, the dosage of corticosteroid (e.g., glucocorticoid) is administered at successively lower dosages with each treatment.Thus, in some such treatment regimens, the dose of corticosteroid is tapered.For example, corticosteroid can be administered at an initial dosage of 4 mg (or equivalent dosage, e.g., for dexamethasone), and at each subsequent administration, the dosage can be reduced so that the dosage is 3 mg for the next administration, 2 mg for the next administration, and 1 mg for the next administration.

[0257] Usually, the dose of corticosteroid administered depends on the specific corticosteroid, since there are differences in potency between different corticosteroids. It will be understood that typically drugs vary in potency and therefore may require different doses to achieve equivalent effect. Table 6 shows the equivalence in potency for various glucocorticoids and routes of administration. Equivalent potency in clinical administration is well known. Information regarding equivalent steroid administration (in a non-chronotherapeutic manner) can be found in the British National Formulary (BNF) 37, March 1999.

[0258] Table 6. Glucocorticoid administration TIFF0007652692000008.tif34160

[0259] Thus, in some embodiments, the steroid is administered at an equivalent dose of 1.0 mg or about 1.0 mg to 20 mg of dexamethasone per day, inclusive, such as 1.0 mg to 15 mg of dexamethasone per day, 1.0 mg to 10 mg of dexamethasone per day, 2.0 mg to 8 mg of dexamethasone per day, or 2.0 mg to 6.0 mg of dexamethasone per day. In some cases, the steroid is administered at an equivalent dose of 4 mg or about 4 mg or 8 mg or about 8 mg of dexamethasone per day.

[0260] In some embodiments, steroids are administered if fever persists after treatment with tocilizumab. For example, in some embodiments, dexamethasone is administered orally or intravenously at a dose of 5-10 mg every 6-12 hours while fever persists. In some embodiments, tocilizumab is administered in conjunction with or after supplemental oxygen.

[0261] B. Microglial Cell Inhibitors In some embodiments, the inhibitor in the combination therapy is an inhibitor of microglial cell activity. In some embodiments, administration of the inhibitor modulates microglial activity. In some embodiments, the inhibitor is an antagonist that inhibits the activity of a signaling pathway in microglia. In some embodiments, the microglial inhibitor affects microglial homeostasis, survival and / or proliferation. In some embodiments, the inhibitor targets the CSF1R signaling pathway. In some embodiments, the inhibitor is an inhibitor of CSF1R. In some embodiments, the inhibitor is a small molecule. In some cases, the inhibitor is an antibody.

[0262] In some aspects, administration of the inhibitor results in one or more effects selected from altering microglial homeostasis and survival, reducing or blocking microglial cell proliferation, reducing or eliminating microglial cells, reducing microglial activation, reducing microglial nitric oxide production, reducing microglial nitric oxide synthase activity, or protecting motor neurons affected by microglial activation. In some embodiments, the agent alters serum or blood levels of biomarkers of CSF1R inhibition, or reduces urinary collagen type 1 cross-linked N-telopeptide (NTX) levels compared to a time point immediately before the start of administration of the inhibitor. In some embodiments, administration of the agent transiently inhibits the activity of microglial activity, and / or the inhibition of microglial activity is not permanent. In some embodiments, administration of the agent transiently inhibits the activity of CSF1R, and / or the inhibition of CSF1R activity is not permanent.

[0263] In some embodiments, the agent that reduces the activity of microglial cells is a small molecule, a peptide, a protein, an antibody or its antigen-binding fragment, an antibody mimic, an aptamer, or a nucleic acid molecule.In some embodiments, the method involves the administration of an inhibitor of microglial activity.In some embodiments, the agent is an antagonist that inhibits the activity of signaling pathways in microglia.In some embodiments, the agent that reduces the activity of microglial cells affects microglial homeostasis, survival, and / or proliferation.

[0264] In some embodiments, the agent that reduces activation of microglial cells is selected from an anti-inflammatory agent, an inhibitor of NADPH oxidase (NOX2), a calcium channel blocker, a sodium channel blocker, inhibits GM-CSF, inhibits CSF1R, specifically binds CSF-1, specifically binds IL-34, inhibits activation of nuclear factor kappa B (NF-κB), inhibits activation of CB 2 Activates receptors and / or CB 2Agonists, which are phosphodiesterase inhibitors, inhibit microRNA-155 (miR-155), upregulate microRNA-124 (miR-124), inhibit nitric oxide production in microglia, inhibit nitric oxide synthase, or activate the transcription factor NRF2 (also called nuclear factor (erythroid-derived 2)-like 2 or NFE2L2).

[0265] In some embodiments, the agent that reduces the activity of microglial cells targets CSF1 (also called macrophage colony-stimulating factor MCSF). In some embodiments, the agent that reduces the activity of microglial cells affects MCSF-stimulated phosphorylation of M-CSF receptor (Pryer et al. Proc Am Assoc Cancer Res, AACR Abstract nr DDT02-2 (2009)). In some cases, the agent that reduces the activity of microglial cells is MCS110 (International Patent Application Publication No. 2014001802; Clinical Trial Study Record No.: A1 NCT00757757; NCT02807844; NCT02435680; NCT01643850).

[0266] In some embodiments, the agent that reduces the activity of microglial cells is a small molecule that targets the CSF1 pathway. In some embodiments, the agent is a small molecule that binds to CSF1R. In some embodiments, the agent is a small molecule that inhibits CSF1R kinase activity by competing with ATP binding to CSF1R kinase. In some embodiments, the agent is a small molecule that inhibits the activation of CFS1R receptor. In some cases, the binding of CSF-1 ligand to CSF1R is inhibited. In some embodiments, the agent that reduces the activity of microglial cells is any of the inhibitors described in US Patent Publication No. 20160032248.

[0267] In some embodiments, the agent is a small molecule inhibitor selected from PLX-3397, PLX7486, JNJ-40346527, JNJ28312141, ARRY-382, PLX73086 (AC-708), DCC-3014, AZD6495, GW2580, Ki20227, BLZ945, PLX647, PLX5622. In some embodiments, the agent is selected from the group consisting of Conway et al., Proc Natl Acad Sci USA, 102(44):16078-83(2005);Dagher et al., Journal of Neuroinflammation, 12:139(2015);Ohno et al., Mol Cancer Ther.5(11):2634-43(2006);von Tresckow et al.Clin Cancer Res.,21(8)(2015);Manthey et al.Mol Cancer Ther.(8(11):3151-61(2009);Pyonteck et al., Nat Med.19(10):1264-1272(2013);Haegel et al., Cancer Res AACR Abstract nr 288(2015);Smith et al., Cancer Res AACR Abstract nr 4889 (2016); Clinical Trial Record Numbers: NCT01525602; NCT02734433; NCT02777710; NCT01804530; NCT01597739; NCT01572519; NCT01054014; NCT01316822; NCT02880371; NCT02673736; any of the inhibitors described in International Patent Application Publication Nos. 2008063888A2, 2006009755A2, U.S. Patent Application Publication Nos. 20110044998, 2014 / 0065141 and 2015 / 0119267.

[0268] In some embodiments, the agent that reduces the activity of microglial cells is 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (BLZ945) or a pharma- ceutically acceptable salt or derivative thereof. In some embodiments, the agent is the following compound: TIFF0007652692000009.tif17128, or a pharma- ceutically acceptable salt thereof, wherein R1 is an alkylpyrazole or alkylcarboxamide, and R2 is a hydroxycycloalkyl.

[0269] In some embodiments, the agent that reduces the activity of microglial cells is 5-((5-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)methyl)-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)pyridin-2-amine, N-[5-[(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)methyl]-2-pyridinyl]-6-(trifluoromethyl)-3-pyridinmethanamine) (PLX 3397) or a pharma- ceutically acceptable salt or derivative thereof. In some embodiments, the agent is 5-(1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-N-[[4-(trifluoromethyl)phenyl]methyl]-2-pyridinamine dihydrochloride (PLX647) or a pharma- ceutically acceptable salt or derivative thereof. In some embodiments, the agent that reduces the activity of microglial cells is the following compound: TIFF0007652692000010.tif32128 or a pharma- ceutically acceptable salt thereof. In some embodiments, the agent that reduces the activity of microglial cells is the following compound: TIFF0007652692000011.tif32128 or a pharma- ceutically acceptable salt thereof. In some embodiments, the agent is any of the inhibitors described in U.S. Patent No. 7,893,075.

[0270] In some embodiments, the agent that reduces the activity of microglial cells is 4-cyano-N-[2-(1-cyclohexen-1-yl)-4-[1-[(dimethylamino)acetyl]-4-piperidinyl]phenyl]-1H-imidazole-2-carboxamide monohydrochloride (JNJ28312141) or a pharma- ceutically acceptable salt or derivative thereof. In some embodiments, the agent is the following compound: TIFF0007652692000012.tif32128 or a pharma- ceutically acceptable salt thereof. In some embodiments, the agent is any of the inhibitors described in U.S. Patent No. 7,645,755.

[0271] In some embodiments, the agent that reduces the activity of microglial cells is 1H-imidazole-2-carboxamide, 5-cyano-N-(2-(4,4-dimethyl-1-cyclohexen-1-yl)-6-(tetrahydro-2,2,6,6-tetramethyl-2H-pyran-4-yl)-3-pyridinyl)-, 4-cyano-1H-imidazole-2-carboxylic acid N-(2-(4,4-dimethylcyclohex-1-enyl)-6-(2,2,6,6-tetramethyltetrahydropyran-4-yl)pyridin-3-yl)amide, 4-cyano-N-(2-(4,4-dimethylcyclohex-1-en-1-yl)-6-(2,2,6,6-tetramethyl-tetrahydro-2H-pyran-4-yl)pyridin-3-yl)-1H-imidazole-2-carboxamide (JNJ-40346527) or a pharma- ceutically acceptable salt or derivative thereof. In some embodiments, the agent is the following compound: TIFF0007652692000013.tif33128 or a pharma- ceutically acceptable salt thereof.

[0272] In another embodiment, the agent that reduces the activity of microglial cells is 5-(3-methoxy-4-((4-methoxybenzyl)oxy)benzyl)pyrimidine-2,4-diamine (GW2580) or a pharma- ceutically acceptable salt or derivative thereof. In some embodiments, the agent is the following compound: TIFF0007652692000014.tif33128, or a pharma- ceutically acceptable salt thereof (International Patent Application Publication No. 2009099553).

[0273] In some embodiments, the agent that reduces the activity of microglial cells is 4-(2,4-difluoroanilino)-7-ethoxy-6-(4-methylpiperazin-1-yl)quinoline-3-carboxamide (AZD6495) or a pharma- ceutically acceptable salt or derivative thereof. In some embodiments, the agent is the following compound: TIFF0007652692000015.tif43128 or a pharma- ceutically acceptable salt thereof.

[0274] In some embodiments, the agent that reduces the activity of microglial cells is N-{4-[(6,7-dimethoxy-4-quinolyl)oxy]-2-methoxyphenyl}-N0-[1-(1,3-thiazol-2-yl)ethyl]urea (Ki20227) or a pharma- ceutically acceptable salt or derivative thereof. In some embodiments, the agent is the following compound: TIFF0007652692000016.tif39128 or a pharma- ceutically acceptable salt thereof.

[0275] In some embodiments, the agent that reduces activation of microglial cells is an antibody that targets the CSF1 pathway. In some embodiments, the agent is an antibody that binds to CSF1R. In some embodiments, the anti-CSF1R antibody blocks dimerization of CSF1R. In some embodiments, the anti-CSF1R antibody blocks the dimerization interface of CSF1R formed by domains D4 and D5 (Ries et al. Cancer Cell 25(6):846-59(2014)). In some cases, the agent is emactuzumab (RG7155; RO5509554), cabilalizumab (FPA-008), LY-3022855 (IMC-CS4), AMG-820, TG-3003, MCS110, H27K15, 12-2D6, 2-4A5 (Rovida and Sbarba, J Clin Cell Immunol. 6:6(2015); Clinical Trial Record Numbers: NCT02760797; NCT01494688; NCT02323191; NCT01962337; NCT02471716; NCT02526017; NCT01346358; NCT02265536; NCT01444404; NCT02713529, NCT00757757; NCT02807844; NCT02435680; NCT01643850).

[0276] In some embodiments, the agent that reduces the activation of microglial cells is a tetracycline antibiotic. For example, the agent affects the concentration of IL-1b, IL-6, TNF-α or iNOS in microglial cells (Yrjaenheikki et al. PNAS 95(26):15769-15774(1998); Clinical Trial Record Number:NCT01120899). In some embodiments, the agent is an opioid antagonist (Younger et al. Pain Med. 10(4):663-672(2009). In some embodiments, the agent reduces glutamatergic neurotransmission (U.S. Pat. No. 5,527,814). In some embodiments, the agent modulates NFkB signaling (Valera et al J. Neuroinflammation 12:93(2015); Clinical Trials Record Number: NCT00231140). In some embodiments, the agent targets cannabinoid receptors (Ramirez et al. J. Neurosci 25(8):1904-13(2005)). In some embodiments, the agent is selected from minocycline, naloxone, riluzole, lenalidomide, and a cannabinoid (optionally WIN55 or 212-2).

[0277] Nitric oxide production by microglia is believed to result in or increase neurotoxicity in some cases. In some embodiments, the agent regulates or inhibits nitric oxide production by microglia. In some embodiments, the agent inhibits nitric oxide synthase (NOS). In some embodiments, the NOS inhibitor is Ronopterin (VAS-203), also known as 4-amino-tetrahydrobiopterin (4-ABH4). In some embodiments, the NOS inhibitor is cindunistat, A-84643, ONO-1714, L-NOARG, NCX-456, VAS-2381, GW-273629, NXN-462, CKD-712, KD-7040 or guanidinoethyl disulfide. In some embodiments, the agent is any of the inhibitors described in Hoeing et al., Cell Stem Cell. 2012 Nov 2;11(5):620-32.

[0278] In some embodiments, the agent blocks, for example, T cell trafficking into the central nervous system. In some embodiments, blocking T cell trafficking can reduce or inhibit immune cells from crossing blood vessel walls into the central nervous system, for example, crossing the blood-brain barrier. In some cases, activated antigen-specific T cells produce proinflammatory cytokines, for example IFN-γ and TNF, which, when reactivated in the CNS, lead to activation of resident cells, for example microglia and astrocytes. See Kivisakk et al., Neurology.2009 Jun 2;72(22):1922-1930. Thus, in some embodiments, isolating activated T cells from microglial cells, for example by blocking trafficking of such cells across the blood-brain barrier and / or inhibiting the ability of such cells to cross the blood-brain barrier, can reduce or eliminate microglial activation. In some embodiments, the agent inhibits adhesion molecules on immune cells, for example T cells. In some embodiments, the agent inhibits integrins. In some embodiments, the integrin is alpha-4 integrin. In some embodiments, the agent is natalizumab (Tysabri®). In some embodiments, the agent modulates a cell surface receptor. In some embodiments, the agent modulates a sphingosine-1-phosphate (S1P) receptor, e.g., S1PR1 or S1PR5. In some embodiments, the agent causes internalization of a cell receptor, e.g., a sphingosine-1-phosphate (S1P) receptor, e.g., S1PR1 or S1PR5. In some embodiments, the agent is fingolimod (Gilenya®) or ozanimod (RPC-1063).

[0279] The transcription factor NRF2 is believed to regulate antioxidant responses, for example, by turning on genes that contain cis-acting elements in their promoter regions. One example of such an element is the antioxidant response element (ARE). In some embodiments, the agent activates NRF2. In some embodiments, activation of NRF2 in microglial cells reduces the responsiveness of the microglial cells to IFN and LPS. In some embodiments, activation of NRF2 inhibits, slows or reduces demyelination, axonal loss, neuronal cell death and / or oligodendrocyte death. In some embodiments, the agent upregulates cytoprotective pathways in cells that are regulated by NRF2. In some embodiments, the agent that activates NRF2 is dimethyl fumarate (Tecfidera®). In some embodiments, the agent is any of the inhibitors described in U.S. Pat. No. 8,399,514. In some embodiments, the agent is any of the inhibitors described in Hoeing et al., Cell Stem Cell. 2012 Nov 2;11(5):620-32.

[0280] In some embodiments, the agent that reduces activation of microglial cells is (4S,4aS,5aR,12aS)-4,7-bis(dimethylamino)-3,10,12,12a-tetrahydroxy-1,11-dioxo-1,4,4a,5,5a,6,11,12a-octahydrotetracene-2-carboxamide (minocycline) or a pharma- ceutically acceptable salt or derivative thereof. In some embodiments, the agent is any of the compounds described in U.S. Patent Application Publication No. 20100190755. In some embodiments, the agent is the following compound: TIFF0007652692000017.tif31128 or a pharma- ceutically acceptable salt thereof.

[0281] In some embodiments, the agent that reduces activation of microglial cells is 3-(7-amino-3-oxo-1H-isoindol-2-yl)piperidine-2,6-dione (lenalidomide) or a pharma- ceutically acceptable salt or derivative thereof. In some embodiments, the agent is the following compound: TIFF0007652692000018.tif33128 or a pharma- ceutically acceptable salt thereof.

[0282] In some embodiments, the agent that reduces activation of microglial cells is 4R,4aS,7aR,12bS)-4a,9-dihydroxy-3-prop-2-enyl-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinolin-7-one (naloxone) or a pharma- ceutically acceptable salt or derivative thereof. In some embodiments, the agent is any of the compounds described in U.S. Pat. No. 8,247,425. In some embodiments, the agent is the following compound: TIFF0007652692000019.tif34128 or a pharma- ceutically acceptable salt thereof.

[0283] In some embodiments, the agent that reduces activation of microglial cells is 2-amino-6-(trifluoromethoxy)benzothiazole, 6-(trifluoromethoxy)benzo[d]thiazol-2-amine or 6-(trifluoromethoxy)-1,3-benzothiazol-2-amine (riluzole) or a pharma- ceutically acceptable salt or derivative thereof, as described in U.S. Patent No. 5,527,814. In some embodiments, the agent is the following compound: TIFF0007652692000020.tif16128 or a pharma- ceutically acceptable salt thereof.

[0284] In some embodiments, the agent that reduces the activation of microglial cells is a modulator of signaling pathways in microglia. In some cases, the agent reduces the signaling of microglia. In some embodiments, the agent is a GM-CSF (CSF2) inhibitor. In other embodiments, the agent that reduces the activation of microglial cells is an ion channel blocker. In some specific embodiments, the agent is a calcium channel blocker. For example, in some specific examples, the agent is a dihydropyridine calcium channel blocker. In some embodiments, the agent is a microRNA inhibitor. For example, the agent targets miR-155. In some embodiments, the agent that reduces microglial cell activation is selected from MOR103, nimodipine, IVIg and LNA-anti-miR-155 (Butoxsky et al. Ann Neurol., 77(1):75-99(2015)and Sanz et al., Br J Pharmacol.167(8):1702-1711(2012); Winter et al., Ann Clin and Transl Neurol.2328-9503(2016); Clinical Trials Record Numbers: NCT01517282, NCT00750867).

[0285] In some embodiments, the agent that reduces activation of microglial cells is 3-(2-methoxyethyl)5-propan-2-yl 2,6-dimethyl-4-(3-nitrophenyl)-1,4-dihydropyridine-3,5-dicarboxylate (nimodipine) or a pharma- ceutically acceptable salt or derivative thereof. In some embodiments, the agent is any of the inhibitors described in U.S. Pat. No. 3,799,934. In some embodiments, the agent is the following compound: TIFF0007652692000021.tif39128 or a pharma- ceutically acceptable salt thereof.

[0286] In some cases, the agent that reduces the activation of microglial cells is administered in a form that only affects the central nervous system and / or does not affect tumor-associated macrophages. In some embodiments, the agent promotes microglial quiescence but does not eliminate or reduce the number of microglia. In some embodiments, the method involves specifically inhibiting microglial activity in the brain, for example as described in Ponomarev et al., Nature Medicine, (1):64-70 (2011).

[0287] Exemplary agents that reduce microglial cell activation and exemplary dosing regimens for administering such agents are shown in Table 7 below.

[0288] Table 7. Exemplary microglial inhibitors and dosing regimens TIFF0007652692000022.tif237166

[0289] C. Other Agents (e.g., Cytokine-Targeting Agents) In some embodiments, an agent (e.g., a toxicity targeting agent) that treats or ameliorates the toxicity of immunotherapy and / or cell therapy, e.g., symptoms of CRS or neurotoxicity, targets a cytokine, e.g., is an antagonist or inhibitor of a cytokine (e.g., transforming growth factor beta (TGF-beta), interleukin 6 (IL-6), interleukin 10 (IL-10), IL-2, MIP1β (CCL4), TNF alpha, IL-1, interferon gamma (IFN-gamma), or monocyte chemoattractant protein-1 (MCP-1)). In some embodiments, the agent that treats or ameliorates the toxicity of immunotherapy and / or cell therapy, e.g., symptoms of CRS or neurotoxicity, targets (e.g., inhibits or is an antagonist of) a cytokine receptor (e.g., IL-6 receptor (IL-6R), IL-2 receptor (IL-2R / CD25), MCP-1 (CCL2) receptor (CCR2 or CCR4), TGF-beta receptor (TGF-beta I, II or III), IFN-gamma receptor (IFNGR), MIP1β receptor (e.g., CCR5), TNF alpha receptor (e.g., TNFR1), IL-1 receptor (IL1-Rα / IL-1Rβ), or IL-10 receptor (IL-10R)).

[0290] The amount of agent that treats or ameliorates the toxicity of selected immunotherapy and / or cell therapy, such as symptoms of CRS or neurotoxicity, administered to ameliorate the symptoms or adverse effects of toxicity, such as symptoms of CRS or neurotoxicity, to immunotherapy and / or cell therapy can be determined by standard clinical methods.Exemplary adverse events include, but are not limited to, increased alanine aminotransferase, increased aspartate aminotransferase, chills, febrile neutropenia, headache, hypotension, left ventricular dysfunction, encephalopathy, hydrocephalus, seizures, and / or tremors.

[0291] In some embodiments, the agent is administered in a dosage of 30 mg or about 30 mg to 5000 mg, e.g., 50 mg to 1000 mg, 50 mg to 500 mg, 50 mg to 200 mg, 50 mg to 100 mg, 100 mg to 1000 mg, 100 mg to 500 mg, 100 mg to 200 mg, 200 mg to 1000 mg, 200 mg to 500 mg, or 500 mg to 1000 mg.

[0292] In some embodiments, the agent is administered at or about 0.5 mg / kg to 100 mg / kg, e.g., at or about 1 mg / kg to 50 mg / kg, 1 mg / kg to 25 mg / kg, 1 mg / kg to 10 mg / kg, 1 mg / kg to 5 mg / kg, 5 mg / kg to 100 mg / kg, 5 mg / kg to 50 mg / kg, 5 mg / kg to 25 mg / kg, 5 mg / kg to 10 mg / kg, 10 mg / kg to 100 mg / kg, 10 mg / kg to 50 mg / kg, 10 mg / kg to 25 mg / kg, 25 mg / kg to 100 mg / kg, 25 mg / kg to 50 mg / kg to 50 mg / kg to 100 mg / kg. In some embodiments, the agent is administered at a dosage of 1 mg / kg or about 1 mg / kg to 10 mg / kg, 2 mg / kg to 8 mg / kg, 2 mg / kg to 6 mg / kg, 2 mg / kg to 4 mg / kg, or 6 mg / kg to 8 mg / kg, inclusive. In some aspects, the agent is administered at a dosage of at least 1 mg / kg, 2 mg / kg, 4 mg / kg, 6 mg / kg, 8 mg / kg, 10 mg / kg or more, or at least about 1 mg / kg, 2 mg / kg, 4 mg / kg, 6 mg / kg, 8 mg / kg, 10 mg / kg or more, or at least about 1 mg / kg, 2 mg / kg, 4 mg / kg, 6 mg / kg, 8 mg / kg, 10 mg / kg or more. In some embodiments, the agent is administered at a dose of 4 mg / kg or 8 mg / kg.

[0293] In some embodiments, the agent is administered by injection, for example, intravenous or subcutaneous injection, intraocular injection, periocular injection, subretinal injection, intravitreal injection, transseptal injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjectval injection, subconjuntival injection, subtenon injection, retrobulbar injection, peribulbar injection, or posterior juxtascleral delivery. In some embodiments, the cells are administered parenterally, intrapulmonary and intranasally, and intralesional administration when localized treatment is desired. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

[0294] In some embodiments, the amount of agent is administered about or approximately twice daily, daily, every other day, three times a week, weekly, biweekly, or monthly.

[0295] In some embodiments, the agent is administered as part of a composition or formulation, such as a pharmaceutical composition or formulation, as described below. Thus, in some cases, a composition containing the agent is administered as described below. In other aspects, the agent is administered alone and can be administered by any known acceptable route of administration, or by those described herein, for example, with respect to compositions and pharmaceutical formulations.

[0296] In some embodiments, the agent that treats or alleviates the toxicity of immunotherapy and / or cell therapy, such as CRS or neurotoxicity symptoms, is an antibody or antigen-binding fragment.In some embodiments, the agent is tocilizumab, siltuximab, sarilumab, olokizumab (CDP6038), elcilimomab, ALD518 / BMS-945429, sirukumab (CNTO 136), CPSI-2634, ARGX-109, FE301 or FM101.

[0297] In some embodiments, the agent is an antagonist or inhibitor of IL-6 or IL-6 receptor (IL-6R). In some aspects, the agent is an antibody that neutralizes the activity of IL-6, such as an antibody or antigen-binding fragment that binds to IL-6 or IL-6R. For example, in some embodiments, the agent is or includes the anti-IL-6R antibody tocilizumab (atlizumab) or sarilumab. In some embodiments, the agent is an anti-IL-6R antibody described in U.S. Pat. No. 8,562,991. In some cases, the agent targeting IL-6 is an anti-IL-6 antibody, such as siltuximab, elcilimomab, ALD518 / BMS-945429, sirukumab (CNTO 136), CPSI-2634, ARGX-109, FE301, FM101, or olokizumab (CDP6038). In some aspects, the agent may neutralize the activity of IL-6 by inhibiting the ligand-receptor interaction. The feasibility of this general type of approach has been demonstrated with natural receptor antagonists for interleukin-1. See Harmurn, CH et al., Nature (1990) 343: 336-340. In some aspects, the IL-6 / IL-6R antagonist or inhibitor is an IL-6 mutein, such as those described in U.S. Pat. No. 5,591,827. In some embodiments, the agent that is an antagonist or inhibitor of IL-6 / IL-6R is a small molecule, a protein or peptide, or a nucleic acid.

[0298] In some embodiments, the agent is tocilizumab. In some embodiments, tocilizumab is administered according to the provided methods and / or as early intervention in the provided articles of manufacture or compositions at a dosage of 1 mg / kg or about 1 mg / kg to 12 mg / kg or about 12 mg / kg, such as 4 mg / kg or about 4 mg / kg, 8 mg / kg or about 8 mg / kg, or 10 mg / kg or about 10 mg / kg. In some embodiments, tocilizumab is administered by intravenous infusion. In some embodiments, tocilizumab is administered for persistent fever of over 39°C that has lasted for 10 hours and is refractory to acetaminophen. In some embodiments, if symptoms return 48 hours after the initial dose, a second dose of tocilizumab is administered.

[0299] In some embodiments, the agent is an agonist or stimulator of TGF-β or a TGF-β receptor (e.g., TGF-β receptor I, II, or III). In some aspects, the agent is an antibody that enhances the activity of TGF-β, such as an antibody or antigen-binding fragment that binds to TGF-β or one of its receptors. In some embodiments, an agent that is an agonist or stimulator of TGF-β and / or its receptor is a small molecule, a protein or peptide, or a nucleic acid.

[0300] In some embodiments, the agent is an antagonist or inhibitor of MCP-1 (CCL2) or MCP-1 receptor (e.g., MCP-1 receptor CCR2 or CCR4). In some aspects, the agent is an antibody that neutralizes the activity of MCP-1, such as an antibody or antigen-binding fragment that binds to MCP-1 or one of its receptors (CCR2 or CCR4). In some embodiments, the antagonist or inhibitor of MCP-1 is any of those described in Gong et al. J Exp Med. 1997 Jul 7; 186(1): 131-137 or Shahrara et al. J Immunol 2008; 180: 3447-3456. In some embodiments, the agent that is an antagonist or inhibitor of MCP-1 and / or its receptor (CCR2 or CCR4) is a small molecule, protein, or peptide, or a nucleic acid.

[0301] In some embodiments, the agent is an antagonist or inhibitor of IFN-γ or IFN-γ receptor (IFNGR). In some aspects, the agent is an antibody that neutralizes the activity of IFN-γ, such as an antibody or antigen-binding fragment that binds to IFN-γ or its receptor (IFNGR). In some aspects, the IFN-gamma neutralizing antibody is any of those described in Dobber et al. Cell Immunol. 1995 Feb; 160(2): 185-92 or Ozmen et al. J Immunol. 1993 Apr 1; ​​150(7): 2698-705. In some embodiments, the agent that is an antagonist or inhibitor of IFN-γ / IFNGR is a small molecule, a protein or peptide, or a nucleic acid.

[0302] In some embodiments, the agent is an antagonist or inhibitor of IL-10 or IL-10 receptor (IL-10R). In some aspects, the agent is an antibody that neutralizes IL-10 activity, such as an antibody or antigen-binding fragment that binds to IL-10 or IL-10R. In some aspects, the IL-10 neutralizing antibody is any of those described in Dobber et al. Cell Immunol. 1995 Feb; 160(2): 185-92 or Hunter et al. J Immunol. 2005 Jun 1; 174(11): 7368-75. In some embodiments, the agent that is an antagonist or inhibitor of IL-10 / IL-10R is a small molecule, a protein or peptide, or a nucleic acid.

[0303] In some embodiments, the agent is an antagonist or inhibitor of IL-1 or IL-1 receptor (IL-1R). In some aspects, the agent is an IL-1 receptor antagonist, which is a modified form of IL-1R, such as anakinra (see, e.g., Fleischmann et al., (2006) Annals of the rheumatic diseases. 65(8):1006-12). In some aspects, the agent is an antibody that neutralizes the activity of IL-1, such as an antibody or antigen-binding fragment that binds to IL-1 or IL-1R, such as canakinumab (see also EP 2277543). In some embodiments, the agent that is an antagonist or inhibitor of IL-1 / IL-1R is a small molecule, a protein or peptide, or a nucleic acid.

[0304] In some embodiments, the agent is an antagonist or inhibitor of tumor necrosis factor (TNF) or tumor necrosis factor receptor (TNFR). In some aspects, the agent is an antibody that blocks the activity of TNF, such as an antibody or antigen-binding fragment that binds to TNF, such as TNFα or its receptor (TNFR, such as TNFRp55 or TNFRp75). In some aspects, the agent is selected from among infliximab, adalimumab, certolizumab pegol, golimumab and etanercept. In some embodiments, the agent that is an antagonist or inhibitor of TNF / TNFR is a small molecule, a protein or peptide, or a nucleic acid. In some embodiments, the agent is a small molecule that affects TNF, such as lenalidomide (see, e.g., Muller et al. (1999) Bioorganic & Medicinal Chemistry Letters. 9(11):1625).

[0305] In some embodiments, the agent is an antagonist or inhibitor of signaling through the Janus kinase (JAK) and two signal transducers and activators of transcription (STAT) signaling cascades. The JAK / STAT proteins are common components of cytokine and cytokine receptor signaling. In some embodiments, the agent is an antagonist or inhibitor of JAK / STAT, such as ruxolitinib (see, e.g., Mesa et al. (2012) Nature Reviews Drug Discovery. 11(2):103-104), tofacitinib (also known as Xeljanz, Jakvinus tasocitinib and CP-690550), baricitinib (LY-3009104, also known as INCB-28050), filgotinib (G-146034, GLPG-0634), gandotinib (LY-2784544), lestaurtinib (CEP-701), momelotinib (GS-0387, CYT-387), pacritinib (SB1518), and upadacitinib (ABT-494). In some embodiments, the agent is a small molecule, a protein or peptide, or a nucleic acid.

[0306] In some embodiments, the agent is a kinase inhibitor. In some embodiments, the agent is an inhibitor of Bruton's tyrosine kinase (BTK). In some embodiments, the inhibitor is or includes ibrutinib or acalabrutinib (see, e.g., Barrett et al., ASH 58 th Annual Meeting San Diego, CA December 3-6, 2016, Abstract 654;Ruella et al., ASH 58 th (See, Annual Meeting San Diego, CA December 3-6, 2016, Abstract 2159). In some embodiments, the agent is an inhibitor described in U.S. Patent Nos. 7,514,444; 8,008,309; 8,476,284; 8,497,277; 8,697,711; 8,703,780; 8,735,403; 8,754,090; 8,754,091; 8,957,079; 8,999,999; 9,125,889; 9,181,257; or 9,296,753.

[0307] In some embodiments, devices such as absorbent resin technology for filtering blood or plasma can be used to reduce cytokine levels. In some embodiments, the device used to reduce cytokine levels is a physical cytokine absorber, such as an extracorporeal cytokine absorber. In some embodiments, a physical cytokine absorber can be used to remove cytokines from the bloodstream in an extracorporeal ex vivo method. In some embodiments, the agent is a porous polymer. In some embodiments, the agent is CytoSorb (see, for example, Basu et al. Indian J Crit Care Med. (2014) 18 (12): 822-824).

[0308] III. Recombinant antigen receptors expressed by cells In some embodiments, cells for use in or administered in connection with the provided methods comprise or are engineered to comprise an engineered receptor, such as an engineered antigen receptor, such as a chimeric antigen receptor (CAR), or a T cell receptor (TCR). Also provided are populations of such cells, compositions comprising such cells, and / or compositions enriched for such cells, such as particular types of cells, such as T cells or CD8 cells. + Or CD4 + Compositions enriched or selected for cells are provided, including pharmaceutical compositions and formulations for administration, e.g., adoptive cell therapy, and therapeutic methods are provided for administering the cells and compositions to a subject, e.g., a patient, according to the methods provided and / or in the articles of manufacture or compositions provided.

[0309] In some embodiments, the cells contain one or more nucleic acids introduced by genetic modification, and thereby express the recombinant or genetically modified products of such nucleic acids.In some embodiments, gene transfer is performed by first stimulating the cells, for example by combining the cells with a stimulus that induces a response, for example proliferation, survival, and / or activation, as measured, for example, by the expression of cytokines or activation markers, followed by transduction of the activated cells and expansion in culture to a number sufficient for clinical application.

[0310] The cells generally express recombinant receptors, such as antigen receptors, including functional non-TCR antigen receptors, such as chimeric antigen receptors (CARs), and other antigen-binding receptors, such as transgenic T cell receptors (TCRs), including other chimeric receptors.

[0311] A. Chimeric Antigen Receptors (CARs) In some embodiments of the provided methods and uses, the chimeric receptor, e.g., chimeric antigen receptor, contains one or more domains that link a ligand binding domain (e.g., an antibody or antibody fragment) that provides specificity for a desired antigen (e.g., a tumor antigen) with an intracellular signaling domain. In some embodiments, the intracellular signaling domain is a stimulatory or activating intracellular domain portion, e.g., a T cell stimulatory domain or a T cell activation domain, that provides a primary activation signal or a primary signal. In some embodiments, the intracellular signaling domain contains or additionally contains a costimulatory signaling domain to promote effector function. In some embodiments, the chimeric receptor, when genetically modified in immune cells, can regulate the activity of T cells, and in some cases, regulate the differentiation or homeostasis of T cells, thereby resulting in genetically engineered cells with improved in vivo longevity, survival, and / or persistence, for example, for use in adoptive cell therapy methods.

[0312] Exemplary antigen receptors, e.g., CARs, as well as methods for modifying and introducing such receptors into cells, are described, for example, in International Patent Application Publication Nos. 200014257, 2013126726, 2012 / 129514, 2014031687, 2013 / 166321, 2013 / 071154, and 2013 / 123061. Nos. 2002131960, 2013287748, 20130149337, U.S. Pat. Nos. 6,451,995, 7,446,190, 8,252,592, 8,339,645, 8,398,282, 7,446,179, 6,410,319, 7,070,995, 7,265,209, 7,354,762, 7,446,191, 8,324,353 and 8,479,118, and European Patent No. 2537416 and / or those described in Sadelain et al., Cancer Discov. 2013 April;3(4):388-398;Davila et al.(2013)PLoS ONE 8(4):e61338;Turtle et al., Curr.Opin.Immunol., 2012 October;24(5):633-39;Wu et al., Cancer, 2012 March 18(2):160-75. In some aspects, antigen receptors include CARs described in U.S. Pat. No. 7,446,190 and those described in International Patent Publication No. 2014055668 A1.Examples of CARs include CARs disclosed in any of the aforementioned publications, such as WO2014031687, US 8,339,645, US 7,446,179, US 2013 / 0149337, U.S. Pat. No. 7,446,190, U.S. Pat. No. 8,389,282, Kochenderfer et al., 2013, Nature Reviews Clinical Oncology, 10, 267-276(2013); Wang et al.(2012) J. Immunother.35(9):689-701; and Brentjens et al., Sci Transl Med.2013 5(177). See also WO2014031687, US 8,339,645, US 7,446,179, US 2013 / 0149337, U.S. Patent No. 7,446,190 and U.S. Patent No. 8,389,282.

[0313] Chimeric receptors (e.g., CARs) generally comprise an extracellular antigen-binding domain, such as a portion of an antibody molecule, generally the variable heavy (V H ) chain region and / or variable light (V L ) chain regions, e.g., scFv antibody fragments.

[0314] In some embodiments, the antigen targeted by the receptor is a polypeptide. In some embodiments, the antigen is a carbohydrate or other molecule. In some embodiments, the antigen is selectively expressed or overexpressed on cells of the disease or condition (e.g., tumor cells or pathogen cells) compared to normal cells or tissues or non-targeted cells or tissues. In other embodiments, the antigen is expressed on normal cells and / or expressed on engineered cells.

[0315] In some embodiments, the antigen targeted by the receptor is αvβ6 integrin (avb6 integrin), B-cell maturation antigen (BCMA), B7-H3, B7-H6, carbonic anhydrase 9 (also known as CA9, CAIX or G250), cancer testis antigen, cancer / testis antigen 1B (CTAG, also known as NY-ESO-1 and LAGE-2), carcinoembryonic antigen (CEA), cyclin, cyclin A2, CC motif chemokine ligand 1 (CCL-1), CD19, CD20, CD22, CD23, CD24, CD30, CD33, CD38, CD44, CD44v6, CD44v7 / 8, CD123, CD133, CD138, CD171, chondroitin sulfate proteoglycan 4 (CSPG4), epidermal growth factor protein (EGFR), epidermal growth factor receptor type III mutant (EGFRvIII), epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), ephrin B2, ephrin receptor A2 (EPHa2), estrogen receptor, Fc receptor-like 5 (FCRL5; also known as Fc receptor homolog 5 or FCRH5), fetal acetylcholine receptor (fetal AchR), folate binding protein (FBP), folate receptor alpha, ganglioside GD2, O-acetylated GD2 (OGD2), ganglioside GD3, glycoprotein Protein 100 (gp100), glypican-3 (GPC3), G protein-coupled receptor class C group 5 member D (GPCR5D), Her2 / neu (receptor tyrosine kinase erb-B2), Her3 (erb-B3), Her4 (erb-B4), erbB dimer, human high molecular weight melanoma-associated antigen (HMW-MAA), hepatitis B surface antigen, human leukocyte antigen A1 (HLA-A1), human leukocyte antigen A2 (HLA-A2), IL-22 receptor alpha ( IL-22Rα), IL-13 receptor alpha 2 (IL-13Rα2), kinase insert domain receptor (kdr), kappa light chain, L1 cell adhesion molecule (L1-CAM), CE7 epitope of L1-CAM, leucine-rich repeat-containing 8 family member A (LRRC8A), Lewis Y, melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, MAGE-A10, mesothelin (MSLN), c-Met, murine cytomegalovirus ( CMV), mucin 1 (MUC1), MUC16, natural killer group 2 member D (NKG2D) ligand, melan A (MART-1), neural cell adhesion molecule (NCAM), oncofetal antigen, preferentially expressed antigen in melanoma (PRAME), progesterone receptor, prostate specific antigen, prostate stem cell antigen (PSCA), prostate specific membrane antigen (PSMA), receptor tyrosine kinase-like orphan receptor 1 (ROR1), survivin, trophoblast glycoprotein (TPBG5T4), tumor associated glycoprotein 72 (TAG72), tyrosinase-related protein 1 (also known as TRP1, TYRP1 or gp75), tyrosinase-related protein 2 (TRP2, also known as dopachrome tautomerase, dopachrome delta-isomerase or DCT), vascular endothelial growth factor receptor (VEGFR), vascular endothelial growth factor receptor 2 (VEGFR2), Wilms' tumor 1 (WT-1), pathogen-specific or pathogen-expressed antigens, or antigens associated with universal tags, and / or biotinylated molecules, and / or molecules expressed by HIV, HCV, HBV or other pathogens, or αvβ6 integrin (avb6 Integrin), B-cell maturation antigen (BCMA), B7-H3, B7-H6, carbonic anhydrase 9 (CA9, also known as CAIX or G250), cancer testis antigen, cancer / testis antigen 1B (CTAG, also known as NY-ESO-1 and LAGE-2), carcinoembryonic antigen (CEA), cyclin, cyclin A2, CC motif chemokine ligand 1 (CCL-1), CD19, CD20, CD22, CD23, CD24, CD30, CD33, CD38, CD44, CD44v6, CD44v7 / 8, CD123, CD133, CD138, CD171, chondroitin sulfate proteoglycan 4 (CSPG4), epidermal growth factor protein (EGFR), epidermal growth factor receptor type III mutation (EGFRvIII), epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), ephrin B2, ephrin receptor A2 (EPHa2), estrogen receptor, Fc receptor-like 5 (FCRL5; also known as Fc receptor homolog 5 or FCRH5), fetal acetylcholine receptor (fetal AchR), folate binding protein (FBP), folate receptor alpha, ganglioside GD2, O-acetylated GD2 (OGD2), ganglioside GD3 , glycoprotein 100 (gp100), glypican-3 (GPC3), G protein-coupled receptor 5D (GPCR5D), Her2 / neu (receptor tyrosine kinase erb-B2), Her3 (erb-B3), Her4 (erb-B4), erbB dimer, human high molecular weight melanoma-associated antigen (HMW-MAA), hepatitis B surface antigen, human leukocyte antigen A1 (HLA-A1), human leukocyte antigen A2 (HLA-A2), IL-22 receptor alpha (IL-22R α), IL-13 receptor alpha 2 (IL-13Rα2), kinase insert domain receptor (kdr), kappa light chain, L1 cell adhesion molecule (L1-CAM), CE7 epitope of L1-CAM, leucine-rich repeat-containing 8 family member A (LRRC8A), Lewis Y, melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, MAGE-A10, mesothelin (MSLN), c-Met, murine cytomegalovirus (CMV) ), mucin 1 (MUC1), MUC16, natural ...

Claims

1. 1. A composition for use in treating a subject having chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL), comprising a dose of engineered T cells, The dose is (i) CD4 containing a chimeric antigen receptor (CAR) that specifically binds to CD19 + and CD8 + Contains T cells; (ii) enriched for CD4+ and CD8+ primary human T cells; (iii) formulated for administration as a plurality of separate compositions, the plurality of separate compositions comprising a CD4 + T cells and CD8 + A first composition comprising one of CD4 T cells and + T cells and CD8 + and a second composition comprising another of the T cells; The dose of engineered T cells comprises a defined ratio of CAR-expressing CD4 + Cells: CAR-expressing CD8 + cells, the ratio being 1:3 to 3:1; and The subject has relapsed following remission following treatment with, has become resistant to, and / or is intolerant to, two or more prior therapies, including a Bruton's tyrosine kinase inhibitor (BTKi) and an inhibitor of Bcl-2. The composition.

2. In the manufacture of a medicament for the treatment of a subject having or suspected of having a disease or condition that is chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL), + and CD8 + 1. Use of a composition of T cells comprising: The composition comprises a dose of engineered T cells, the dose comprising: (i) CD4 containing a chimeric antigen receptor (CAR) that specifically binds to CD19 + and CD8 + Contains T cells; (ii) enriched for CD4+ and CD8+ primary human T cells; (iii) formulated for administration as a plurality of separate compositions, the plurality of separate compositions comprising a CD4 + T cells and CD8 + A first composition comprising one of CD4 T cells and + T cells and CD8 + and a second composition comprising another of the T cells; The dose of engineered T cells comprises a defined ratio of CAR-expressing CD4 + Cells: CAR-expressing CD8 + cells, the ratio being 1:3 to 3:1; and The subject has relapsed following remission following treatment with, has become resistant to, and / or is intolerant to, two or more prior therapies, including a Bruton's tyrosine kinase inhibitor (BTKi) and an inhibitor of Bcl-2. The above uses.

3. 2. The composition of claim 1, wherein the BTKi is ibrutinib.

4. 3. The use of claim 2, wherein the BTKi is ibrutinib.

5. 1. A composition for use in treating a subject having or suspected of having chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL), comprising a dose of engineered T cells, (i) the dose comprises a CD4 + and CD8 + Contains T cells; (ii) the composition is formulated for administration as a plurality of separate compositions, the plurality of separate compositions comprising a plurality of separate compositions each comprising a CD4 + T cells and CD8 + A first composition comprising one of T cells and CD4 + T cells and CD8 + and a second composition comprising another of the T cells; (iii) the dose of engineered T cells is a CD4 T cell expressing a CAR in a defined ratio; + Cells: CAR-expressing CD8 + cells, the ratio being 1:3 to 3:1; and (iv) the subject has relapsed following remission following treatment with, has become resistant to, and / or is intolerant to, two or more prior therapies, including a Bruton's tyrosine kinase inhibitor (BTKi) and an inhibitor of Bcl-2. The composition.

6. In the manufacture of a medicament for the treatment of a subject having or suspected of having a disease or condition that is chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL), + and CD8 + 1. Use of a composition of T cells, the composition comprising a dose of engineered T cells, (i) the dose comprises a CD4 + and CD8 + Contains T cells; (ii) the composition is formulated for administration as a plurality of separate compositions, the plurality of separate compositions comprising a plurality of separate compositions each comprising a CD4 + T cells and CD8 + A first composition comprising one of T cells and CD4 + T cells and CD8 + and a second composition comprising another of the T cells; (iii) the dose of engineered T cells is a CD4 T cell expressing a CAR in a defined ratio; + Cells: CAR-expressing CD8 + cells, the ratio being 1:3 to 3:1; and (iv) the subject has relapsed following remission following treatment with, has become resistant to, and / or is intolerant to, two or more prior therapies, including a Bruton's tyrosine kinase inhibitor (BTKi) and an inhibitor of Bcl-2. The above uses.

7. The dose of engineered T cells is a 1:1 ratio of CD4 + Cells: CAR-expressing CD8 + 6. The composition of any one of claims 1, 3, and 5, comprising a cell.

8. The composition of any one of claims 1, 3, 5, and 7, wherein the dose of engineered T cells comprises from 2.5 x 107 total CAR-expressing cells to 1.5 x 108 total CAR-expressing cells.

9. 9. The composition of any one of claims 5, 7, and 8, wherein said dose of engineered T cells is enriched for CD4+ and CD8+ primary human T cells.

10. 10. The composition of any one of claims 1, 3, 5, and 7-9, wherein said dose of engineered T cells enriched for primary human CD4+ and CD8+ T cells comprises greater than 70%, greater than 75%, greater than 80%, greater than 85%, greater than 90%, greater than 95%, or greater than 98% CD4+ and CD8+ primary human T cells.

11. said dose of engineered T cells (a) 2.5×107 total CAR-expressing cells to 1.0×108 total CAR-expressing cells; and / or (b) 2.5×107 total CAR-expressing cells; 5×107 total cells or total CAR-expressing cells; or 1 x 108 total cells or total CAR expressing cells The composition of any one of claims 1, 3, 5, and 7-10, comprising:

12. CD4 + CAR and / or CD8 contained by T cells + The CAR contained by the T cell contains the same CAR and / or is a CD4 + T cells and / or CD8 + The composition of any one of claims 1, 3, 5, and 7-11, wherein the T cells are genetically engineered to express the same CAR.

13. The first composition is a CD8 + The second composition comprises CD4 T cells. + The composition of any one of claims 1, 3, 5, and 7 to 12, comprising T cells.

14. 14. The composition of claim 13, wherein the first composition is to be administered prior to the second composition.

15. The first composition and the second composition comprise: (a) For administration at intervals of 48 hours or less; (b) For administration at intervals of 36 hours or less, 24 hours or less, 12 hours or less, 6 hours or less, 4 hours or less, 2 hours or less, 1 hour or less, or 30 minutes or less; (c) for administration on the same day, at intervals of 0 to 12 hours, 0 to 6 hours, or 0 to 2 hours; (d) for administration at intervals of 1 minute to 1 hour or 5 minutes to 30 minutes; or (e) For administration separated by 2 hours or less, 1 hour or less, 30 minutes or less, 15 minutes or less, 10 minutes or less, or 5 minutes or less; The composition of any one of claims 1, 3, 5, and 7 to 14.

16. the subject has or is suspected of having CLL; or The subject is identified or selected as having CLL. The composition of any one of claims 1, 3, 5, and 7 to 15.

17. The subject has or is suspected of having SLL; or The subject is identified or selected as having SLL, The composition of any one of claims 1, 3, 5, and 7 to 16.

18. The composition of any one of claims 1, 3, 5, and 7-17, wherein the two or more prior therapies further comprise a combination therapy comprising fludarabine and rituximab; radiation therapy; or hematopoietic stem cell transplant (HSCT).

19. The composition of any one of claims 1, 3, 5, and 7-18, wherein the inhibitor of Bcl-2 is venetoclax.

20. The composition of any one of claims 1, 3, 5, and 7-19, wherein the BTKi is ibrutinib and the inhibitor of Bcl-2 is venetoclax.

21. At or prior to use of the composition: (a) the subject is identified or has been identified as having one or more cytogenetic abnormalities; (b) the subject is identified or has been identified as having standard risk CLL or SLL; (c) the subject (1) is identified or has been identified as intolerant to a Bruton's tyrosine kinase (BTK) inhibitor and has received a BTK inhibitor for less than six months, or (2) is ineligible for treatment with a BTK inhibitor.

21. The composition of any one of claims 1, 3, 5, and 7 to 20.

22. 22. The composition of claim 21, wherein the one or more cytogenetic abnormalities are selected from among a complex karyotype or cytogenetic abnormality, del 17p, an unmutated IGVH gene, and a TP53 mutation.

23. (a) the subject is identified or identified as having an ECOG status of 0 or 1 and / or the subject does not have an ECOG status of >1; (b) the subject does not have Richter's transformation of CLL or SLL at or immediately prior to the use of the composition or at or immediately prior to the administration of lymphodepletion therapy; (c) the subject is an adult and / or is over 50, 60, or 70 years of age; (d) the engineered T cells are autologous to the subject; (e) prior to use of the composition, the subject is administered lymphodepleting therapy; and / or (f) the use of the composition and / or administration of lymphodepletion therapy is via outpatient delivery; 23. The composition of any one of claims 1, 3, 5, and 7 to 22.

24. The lymphodepletion therapy comprises: (i) administration of fludarabine and / or cyclophosphamide; (ii) 200 to 400 mg / m 2 (inclusive) cyclophosphamide and / or 20–40 mg / m 2 of fludarabine daily for 2 to 4 days; and / or (iii) 300 mg / m 2 of cyclophosphamide and 30 mg / m 2 of fludarabine daily for three days.

24. The composition of claim 23, comprising:

25. 25. The composition of claim 24, wherein said dose of cells is administered at least 2-7 days after lymphodepletion therapy or at least 2-7 days after initiation of lymphodepletion therapy.

26. (a) a response in at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of treated subjects is the objective response rate; (b) the response in at least 35%, at least 40%, at least 50%, at least 60%, or at least 70% of treated subjects is a complete response (CR) with a duration of response until progression lasting greater than 3 months or greater than 6 months; (c) greater than 50%, greater than 60%, or greater than 70% had undetectable minimal residual disease (MRD) for at least 1 month, at least 2 months, at least 3 months, or at least 6 months following administration of the dose of engineered T cells; or (d) 10% or less of subjects exhibit cytokine release syndrome (CRS) greater than Grade 2; and / or (e) 10% or less, 20% or less, 30% or less, or 40% or less of subjects experience neurotoxicity greater than Grade 2; 26. The composition of any one of claims 1, 3, 5, and 7 to 25.

27. 7. The use of any one of claims 2, 4 and 6, wherein said dose of engineered T cells is enriched for CD4+ and CD8+ primary human T cells.

28. The use of any one of claims 2, 4, 6, and 27, wherein the dose of engineered T cells comprises from 2.5x107 total CAR-expressing cells to 1.5x108 total CAR-expressing cells.

29. The first composition is a CD8 + The second composition comprises CD4 T cells. + 30. The use according to any one of claims 2, 4, 6, 27 and 28, comprising T cells.

30. The use of any one of claims 2, 4, 6, and 27-29, wherein the two or more previous therapies further comprise a combination therapy comprising fludarabine and rituximab; radiation therapy; or hematopoietic stem cell transplantation (HSCT).

31. The use according to any one of claims 2, 4, 6 and 27 to 30, wherein the inhibitor of Bcl-2 is venetoclax.

32. The use of any one of claims 2, 4, 6 and 27 to 31, wherein the BTKi is ibrutinib and the inhibitor of Bcl-2 is venetoclax.

33. CAR, an extracellular antigen-binding domain specific for CD19; A transmembrane domain; a cytoplasmic signaling domain derived from a costimulatory molecule; Cytoplasmic signaling domains derived from primary signaling ITAM-containing molecules and / or the CAR, in turn, comprises an extracellular antigen-binding domain specific for CD19, a transmembrane domain, a cytoplasmic signaling domain derived from a costimulatory molecule, and a cytoplasmic signaling domain derived from a primary signaling ITAM-containing molecule; 27. The composition of any one of claims 1, 3, 5, and 7 to 26.

34. The composition of claim 33, wherein the costimulatory molecule is the signaling domain of 4-1BB and the primary signaling ITAM-containing molecule is CD3ζ.

35. The composition of claim 33 or 34, wherein the antigen-binding domain is an scFv.

36. The scFv comprises a CDRL1 sequence of RASQDISKYLN (SEQ ID NO: 35), a CDRL2 sequence of SRLHSGV (SEQ ID NO: 36), and a CDRL3 sequence of GNTLPYTFG (SEQ ID NO: 37), and a CDRH1 sequence of DYGVS (SEQ ID NO: 38), a CDRH2 sequence of VIWGSETTYYNSALKS (SEQ ID NO: 39), and a CDRH3 sequence of YAMDYWG (SEQ ID NO: 40); the scFv comprises the variable heavy (VH) chain region of FMC63 and the variable light (VL) chain region of FMC63, and / or the CDRL1 sequence of FMC63, the CDRL2 sequence of FMC63, the CDRL3 sequence of FMC63, the CDRH1 sequence of FMC63, the CDRH2 sequence of FMC63, and the CDRH3 sequence of FMC63, or binds to the same epitope as any of the foregoing or competes for binding with any of the foregoing; The scFv comprises a VH as set forth in SEQ ID NO: 41 and a VL as set forth in SEQ ID NO: 42; 36. The composition of claim 35.

37. the scFv comprises a variable heavy (VH) chain region and a variable light (VL) chain region, the VH and VL being separated by a flexible linker, the flexible linker being or comprising the sequence shown in SEQ ID NO:24; and / or The scFv is or comprises the sequence shown in SEQ ID NO:

43.

37. The composition of claim 35 or 36.

38. (a) the cytoplasmic signaling domain derived from the costimulatory molecule is the signaling domain of CD28 or 4-1BB; and / or (b) the cytoplasmic signaling domain derived from the primary signaling ITAM-containing molecule is a CD3 zeta signaling domain; The composition according to any one of claims 33 to 37.

39. (i) the cytoplasmic signaling domain derived from the costimulatory molecule is the signaling domain of 4-1BB; and / or (ii) the cytoplasmic signaling domain from the costimulatory molecule comprises SEQ ID NO: 12, or a variant thereof having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto; 39. The composition of claim 38.

40. 40. The composition of claim 38 or 39, wherein the cytoplasmic signaling domain from the primary signaling ITAM-containing molecule comprises an amino acid sequence having SEQ ID NO: 13 or 14 or 15, or at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto.

41. The composition of any one of claims 33 to 40, wherein the transmembrane domain comprises the transmembrane portion of CD28.

42. 42. The composition of any one of claims 33-41, wherein the transmembrane domain comprises SEQ ID NO:8 or 9, or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identity thereto.

43. The composition of any one of claims 35 to 37, wherein the CAR further comprises a spacer between the transmembrane domain and the scFv.

44. (a) the spacer is a polypeptide spacer comprising or consisting of all or a portion of an immunoglobulin hinge or a modified version thereof; (b) the spacer is 15 amino acids or less and does not include the CD28 extracellular domain and the CD8 extracellular domain; (c) the spacer is 12 amino acids in length; or (d) a spacer, having or consisting of a sequence encoded by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, or a variant of any of the foregoing having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto; and / or X 1 is glycine, cysteine, or arginine and X 2 is cysteine ​​or threonine; 1 PPX 2 comprising or consisting of P, 44. The composition of claim 43.

45. 45. The composition of claim 44, wherein the immunoglobulin hinge or modified version thereof is an IgG4 hinge or modified version thereof.

46. The composition of any one of claims 1, 3, 5, 7-26, and 33-45, wherein the subject is a human subject.

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