Treatment of myasthenia gravis using a population of CD19 car-expressing cells

Abstract

The invention provides methods of using a population of CD19 CAR-expressing cells (for example, rapcabtagene autoleucel) for treating myasthenia gravis. Also disclosed are methods of making a population of CD19 CAR-expressing cells.

Description

[0001] PAT059889-PCT-SEC01

[0002] TREATMENT OF MYASTHENIA GRAVIS USING A POPULATION OF CD 19 CAR¬

[0003] EXPRESSING CELLS

[0004] FIELD OF THE INVENTION

[0005] The present invention relates generally to methods of using a population of CD 19 CAR- expressing cells for treating myasthenia gravis, compositions comprising the same, and methods of making CD 19 CAR-expressing cells.

[0006] BACKGROUND OF THE INVENTION

[0007] Myasthenia gravis (MG) is a prototypical autoimmune disease in which autoantibodies target the neuromuscular junction (NMJ), causing chronic, fluctuating, and often debilitating weakness and muscle fatigability. Patients with MG whose disease does not respond to current (typically immunosuppressive) therapies or who have serious side-effects associated with their use represent a substantial unmet medical need. Autoantibody-producing B cells and plasma cells are key cellular components of MG pathophysiology.

[0008] Beside symptomatic therapy with acetylcholinesterase inhibitors, immune therapy is standard of care (SOC) in gMG The treatment has evolved from nonspecific immunosuppression using corticosteroids and drugs like azathioprine to newly licensed targeted interventions blocking C5 complement or the FcRn. However, although these new modalities are now available, there is still a proportion (5-10%) of patients that do not respond sufficiently to these treatments or have unacceptable side effects limiting their use. Furthermore, all these treatments typically require long-term administration which may be associated with significant risks and side effects. There is no available therapy that has been shown to reliably induce longterm remission (possible exception of thymectomy), free of immunomodulating drugs. Thus, there is a high unmet need for a treatment that addresses the underlying MG pathophysiology to slow or completely stop disease progression.

[0009] SUMMARY OF THE INVENTION

[0010] Disclosed herein are methods of using a population of CD 19 CAR-expressing cells for treating myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, PAT059889-PCT-SEC01 antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, in a subject. The present disclosure also pertains to methods of making CD 19 CAR-expressing cells, and compositions generated using such methods.

[0011] In one aspect, the disclosure provides a method of treating a subject having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, the method comprising administering to the subject a population of cells engineered to express a CD 19 CAR (“a population of CD19 CAR-expressing cells”), said population comprising:

[0012] (a) about the same percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;

[0013] (b) a change within about 5% to about 10% of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, for example, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;

[0014] (c) an increased percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, for example, increased by at least 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, or 3-fold, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;

[0015] (d) about the same percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0016] (e) a change within about 5% to about 10% of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, as compared to the percentage of central memory cells, for example, central memory T cells, for example, PAT059889-PCT-SEC01

[0017] CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0018] (f) a decreased percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, for example, decreased by at least 20, 25, 30, 35, 40, 45, or 50%, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0019] (g) about the same percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0020] (h) a change within about 5% to about 10% of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR; or

[0021] (i) an increased percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR.

[0022] In one aspect, the disclosure provides a method of treating a subject having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, the method comprising administering to the subject a population of cells engineered to express a CD 19 CAR (“a population of CD 19 CAR-expressing cells”), wherein:

[0023] (a) the median GeneSetScore (Up TEM vs. Down TSCM) of the population of cells is about the same as or differs by no more than (for example, increased by no more than) about 25, 50, 75, 100, or 125% from the median GeneSetScore (Up TEM vs. Down TSCM) of the same population of cells prior to being engineered to express the CAR;

[0024] (b) the median GeneSetScore (Up Treg vs. Down Teff) of the population of cells is about the same as or differs by no more than (for example, increased by no more than) about 25, 50, PAT059889-PCT-SEC01

[0025] 100, 150, or 200% from the median GeneSetScore (Up Treg vs. Down Teff) of the population of cells prior to being engineered to express the CAR;

[0026] (c) the median GeneSetScore (Down sternness) of the population of cells is about the same as or differs by no more than (for example, increased by no more than) about 25, 50, 100, 150, 200, or 250% from the median GeneSetScore (Down sternness) of the population of cells prior to being engineered to express the CAR;

[0027] (d) the median GeneSetScore (Up hypoxia) of the population of cells is about the same as or differs by no more than (for example, increased by no more than) about 125, 150, 175, or 200% from the median GeneSetScore (Up hypoxia) of the population of cells prior to being engineered to express the CAR; or

[0028] (e) the median GeneSetScore (Up autophagy) of the population of cells is about the same as or differs by no more than (for example, increased by no more than) about 180, 190, 200, or 210% from the median GeneSetScore (Up autophagy) of the population of cells prior to being engineered to express the CAR.

[0029] In one aspect, the disclosure provides a method of treating a subject having an myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, the method comprising administering to the subject rapcabtagene autoleucel.

[0030] In one aspect, the disclosure provides a method of treating a subject myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, the method comprising administering to the subject a population of CD 19 CAR-expressing cells in an amount sufficient to treat the neuro immune disease. In some embodiments, the neuroimmune disease is myasthenia gravis. In some embodiments, the population of CD 19 CAR-expressing cells is rapcabtagene autoleucel.

[0031] In some embodiments, the population of CD 19 CAR-expressing cells is administered at a dose of about 0.5 x 106to 90 x 106viable CAR-expressing cells, optionally wherein the population of CD 19 CAR-expressing cells is administered at a dose of 12.5 x 106viable CAR- expressing cells, optionally wherein the population of CAR-expressing cells is administered at a dose of 7.5 x 106viable CAR-expressing cells, optionally wherein the population of CAR- PAT059889-PCT-SEC01 expressing cells is administered at a dose of 5 x 106viable CAR-expressing cells, optionally wherein the population of CAR-expressing cells is administered at a dose of 2.5 x 106viable CAR-expressing cells. In some embodiments, the population of CD 19 CAR-expressing cells is rapcabtagene autoleucel.

[0032] In some embodiments, the population of CD 19 CAR-expressing cells is administered at a dose of about 2.5 x 106to 2.5 x 108viable CAR-expressing cells, for example, about 12.5 x 106viable CAR-expressing cells. In some embodiments, the population of CD19 CAR-expressing cells is rapcabtagene autoleucel.

[0033] In some embodiments, the population of CD 19 CAR-expressing cells is administered at a dose of about 12.5 x 106to 1.25 x 109viable CAR-expressing cells, for example, about 25 x 106viable CAR-expressing cells. In some embodiments, the population of CD19 CAR-expressing cells is rapcabtagene autoleucel.

[0034] In some embodiments, the population of CD 19 CAR-expressing cells is administered at a dose of about 2.5 x 106to 2.5 x 108viable CAR-expressing cells, for example, about 40 x 106viable CAR-expressing cells. In some embodiments, the population of CD19 CAR-expressing cells is rapcabtagene autoleucel.

[0035] In one aspect, the disclosure provides a population of CD 19 CAR-expressing cells or a pharmaceutical composition comprising the same for use in a method of modulating an immune response in a subject having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, said method comprising administering to the subject an effective amount of the population of CD 19 CAR-expressing cells or an effective amount of the pharmaceutical composition. In some embodiments, the population of CD 19 CAR-expressing cells is rapcabtagene autoleucel.

[0036] In one aspect, the disclosure provides a method of treating a subject having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, the method comprising administering to the subject: PAT059889-PCT-SEC01 a population of cells that express, or comprise a nucleic acid configured to express, a CD 19 chimeric antigen receptor (CD 19 CAR), optionally wherein the population of cells is rapcabtagene autoleucel, and a second therapy, wherein the second therapy and CD 19 CAR cells are present in the subject at the same time, e.g., wherein the second therapy is administered at a time when the CD 19 CAR cells are present in the subject.

[0037] In one aspect, the disclosure provides rapcabtagene autoleucel or a pharmaceutical composition comprising the same for use in a method of modulating an immune response in a subject having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, said method comprising administering to the subject an effective amount of the population of rapcaptag ene autoleucel or an effective amount of the pharmaceutical composition

[0038] In one aspect, the disclosure provides rapcabtagene autoleucel or a pharmaceutical composition comprising the same for use in a method of modulating an immune response in a subject having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, said method comprising administering to the subject an effective amount of rapcabtagene autoleucel or an effective amount of the pharmaceutical composition.

[0039] Rapcabtagene autoleucel for use in the manufacture of a medicament for treating a subject having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies.

[0040] Rapcabtagene autoleucel for use in the manufacture of a medicament for treating a subject having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, wherein rapcabtagene autoleucel is formulated for administration in an amount sufficient to treat the myasthenia gravis. PAT059889-PCT-SEC01

[0041] Rapcabtagene autoleucel for use in the manufacture of a medicament for treating a subject having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, rapcabtagene autoleucel is formulated for administration at a dose of 0.5 - 50 x 106viable CAR+ T cells (e.g., 2.5-, 5-, 7.5-, 12.5-, 25-, or 40 x 106viable CAR+ T cells).

[0042] Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references (for example, sequence database reference numbers) mentioned herein are incorporated by reference in their entirety. For example, all GenBank, Unigene, and Entrez sequences referred to herein, for example, in any Table herein, are incorporated by reference. When one gene or protein references a plurality of sequence accession numbers, all of the sequence variants are encompassed.

[0043] In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. Headings, sub-headings or numbered or lettered elements, for example, (a), (b), (i) etc., are presented merely for ease of reading. The use of headings or numbered or lettered elements in this document does not require the steps or elements be performed in alphabetical order or that the steps or elements are necessarily discrete from one another. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0044] BRIEF DESCRIPTION OF THE FIGURES

[0045] FIG. 1 is a schematic showing the clinical trial design for a phase 1 / 2 study, open-label, multi-center, to assess safety, efficacy and cellular kinetics of ARM-CD19 CAR T cells in participants with myasthenia gravis.

[0046] FIG. 2 is a schematic showing the cohort design.

[0047] FIG. 3 is a schematic comparing the ARM process to a traditional CAR T manufacturing process. PAT059889-PCT-SEC01

[0048] DETAILED DESCRIPTION

[0049] Definitions

[0050] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains.

[0051] The term “a” and “an” refers to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0052] The term “about” when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20% or in some instances ±10%, or in some instances ±5%, or in some instances ±1%, or in some instances ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.

[0053] The term “Chimeric Antigen Receptor” or alternatively a “CAR” refers to a recombinant polypeptide construct comprising at least an extracellular antigen-binding domain, a transmembrane domain and a cytoplasmic signaling domain (also referred to herein as “an intracellular signaling domain”) comprising a functional signaling domain derived from a stimulatory molecule as defined below. In some embodiments, the domains in the CAR polypeptide construct are in the same polypeptide chain, for example, comprise a chimeric fusion protein.

[0054] The term “autologous” refers to any material derived from the same individual to whom it is later to be re-introduced into the individual.

[0055] The term “allogeneic” refers to any material derived from a different animal of the same species as the individual to whom the material is introduced. Two or more individuals are said to be allogeneic to one another when the genes at one or more loci are not identical. In some embodiments, allogeneic material from individuals of the same species may be sufficiently unlike genetically to interact antigenically.

[0056] The term “apheresis” as used herein refers to the art-recognized extracorporeal process by which the blood of a donor or patient is removed from the donor or patient and passed through an apparatus that separates out selected constituent(s) and returns the remainder to the circulation of PAT059889-PCT-SEC01 the donor or patient, for example, by re- transfusion. Thus, in the context of “an apheresis sample” refers to a sample obtained using apheresis.

[0057] “Immune effector cell,” as that term is used herein, refers to a cell that is involved in an immune response, for example, in the promotion of an immune effector response. Examples of immune effector cells include T cells, for example, alpha / beta T cells and gamma / delta T cells, B cells, natural killer (NK) cells, natural killer T (NKT) cells, mast cells, and myeloid-derived phagocytes.

[0058] “Immune effector function or immune effector response,” as that term is used herein, refers to function or response, for example, of an immune effector cell, that enhances or promotes an immune attack of a target cell. For example, an immune effector function or response refers a property of a T or NK cell that promotes killing or the inhibition of growth or proliferation, of a target cell. In the case of a T cell, primary stimulation and costimulation are examples of immune effector function or response.

[0059] The term “effector function” refers to a specialized function of a cell. Effector function of a T cell, for example, may be cytolytic activity or helper activity including the secretion of cytokines.

[0060] The term “effective amount” or “therapeutically effective amount” are used interchangeably herein, and refer to an amount of a compound, formulation, material, or composition, as described herein effective to achieve a particular biological result.

[0061] The term “endogenous” refers to any material from or produced inside an organism, cell, tissue, or system.

[0062] The term “exogenous” refers to any material introduced from or produced outside an organism, cell, tissue, or system.

[0063] The term “expression” refers to the transcription and / or translation of a particular nucleotide sequence. In some embodiments, expression comprises translation of an mRNA introduced into a cell.

[0064] The term “lentivirus” refers to a genus of the Retroviridae family. Lentiviruses are unique among the retroviruses in being able to infect non-dividing cells; they can deliver a significant amount of genetic information into the DNA of the host cell, so they are one of the most efficient methods of a gene delivery vector. HIV, SIV, and FIV are all examples of lentiviruses. PAT059889-PCT-SEC01

[0065] The term “lentiviral vector” refers to a vector derived from at least a portion of a lentivirus genome, including especially a self-inactivating lentiviral vector as provided in Milone et al., Mol. Ther. 17(8): 1453-1464 (2009). Other examples of lentivirus vectors that may be used in the clinic, include but are not limited to, for example, the LENUVECTOR® gene delivery technology from Oxford BioMedica, the LENUMAX™ vector system from Lentigen and the like. Nonclinical types of lentiviral vectors are also available and would be known to one skilled in the art.

[0066] The term “parenteral” administration of an immunogenic composition includes, for example, subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrasternal injection, intratumoral, or infusion techniques.

[0067] As used herein, “B cell antigen” refers to an antigen associated with a B cell. Nonlimiting examples of molecules associated with a B cell include proteins expressed on the surface of B cells, e.g. CD19, BCMA, CD22, CD20, CD10, CD34, CD123, FLT-3, ROR1, CD79b, CD 179b, or CD79a.

[0068] As used herein, the term “CD 19” refers to the Cluster of Differentiation 19 protein. The human and murine amino acid and nucleic acid sequences can be found in a public database, such as GenBank, UniProt and Swiss-Prot. For example, the amino acid sequence of human CD19 can be found as UniProt / Swiss-Prot Accession No. Pl 5391 and the nucleic acid sequence encoding of the human CD19 can be found at Accession No. NM 001178098. It is also an early marker of B cell progenitors. See, e.g., Nicholson et al. Mol. Immun. 34 (16-17): 1157-1165 (1997). In one aspect the antigen-binding portion of the CART recognizes and binds an antigen within the extracellular domain of the CD 19 protein. In one aspect, the CD 19 protein is expressed on an autoreactive B-cell. As used herein, “CD 19” includes proteins comprising mutations, e.g., point mutations, fragments, insertions, deletions and splice variants of full length wild-type CD 19.

[0069] As used herein, the terms “treat”, “treatment” and “treating” refer to the reduction or amelioration of the progression, severity and / or duration of an neuroimmune disorder, or the amelioration of one or more symptoms (preferably, one or more discernible symptoms) of an neuroimmune disorder resulting from the administration of one or more therapies (for example, one or more therapeutic agents such as a CAR of the invention). In specific embodiments, the terms “treat,” “treatment,” and “treating” refer to the amelioration of at least one measurable PAT059889-PCT-SEC01 physical parameter of a neuroimmune disorder, such as autoantibodies, not necessarily discernible by the patient. In other embodiments the terms “treat”, “treatment” and “treating” refer to the inhibition of the progression of a neuroimmune disorder, either physically by, for example, stabilization of a discernible symptom, physiologically by, for example, stabilization of a physical parameter, or both.

[0070] The term “subject” is intended to include living organisms in which an immune response can be elicited (for example, mammals, for example, human).

[0071] The term “therapeutic” as used herein means a treatment. A therapeutic effect is obtained by reduction, suppression, remission, or eradication of a disease state.

[0072] The term “prophylaxis” as used herein means the prevention of or protective treatment for a disease or disease state.

[0073] “Refractory” as used herein refers to a neuroimmune disease or disorder, for example, myasthenia gravis, which does not respond to a treatment. In embodiments, a refractory neuroimmune disease or disorder can be resistant to a treatment before or at the beginning of the treatment. In other embodiments, the refractory neuroimmune disease or disorder can become resistant during a treatment.

[0074] As used herein, “myasthenia gravis” refers to all types and manifestations of myasthenia gravis. Manifestations of myasthenia gravis include, without limitation, early-onset myasthenia gravis, late-onset myasthenia gravis, thyoma-associated myasthenia gravis, antibody mediated myasthenia gravis (e.g., AChR+, MuSK+, LRP4+), ocular myasthenia gravis, or myasthenia gravis with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies.

[0075] As used herein, “severe refractory neuroimmune disease” refers to a manifestation of an neuroimmune disease that has failed to respond (e.g., remains charactericterized by high disease activity) following at least one standard immunosuppressive therapy or at least one biological agent.

[0076] “Relapsed” or “relapse” as used herein refers to the return or reappearance of a disease (for example, a neuroimmune disease or disorder) or the signs and symptoms of a disease such as a neuroimmune disease or disorder after a period of improvement or responsiveness, for example, after prior treatment of a therapy, for example, standard of care therapy. The initial period of responsiveness may involve the level of autoantibodies cells falling below a certain PAT059889-PCT-SEC01 threshold. The reappearance may involve the level of autoantibodies rising above a certain threshold.

[0077] “Remission” as used herein refers to a decrease in or disappearance of signs and symptoms of a disease (for example, a neuroimmune disease or disorder). Remission may be partial or complete. In partial remission, some, but not all, signs and symptoms of a disease have decreased or disappeared. In complete remission, all signs and symptoms of a disease have disappeared. Remission may be determined according to a defined set of criteria established for a particular disease (for example, a neuroimmune disease or disorder).

[0078] Ranges: throughout this disclosure, various embodiments of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. As another example, a range such as 95-99% identity, includes something with 95%, 96%, 97%, 98%, or 99% identity, and includes subranges such as 96-99%, 96-98%, 96- 97%, 97-99%, 97-98%, and 98-99% identity. This applies regardless of the breadth of the range.

[0079] Administered “in combination”, as used herein, means that two (or more) different treatments are delivered to the subject during the course of the subject's affliction with the disorder, for example, the two or more treatments are delivered after the subject has been diagnosed with the disorder and before the disorder has been cured or eliminated or treatment has ceased for other reasons. In some embodiments, the delivery of one treatment is still occurring when the delivery of the second begins, so that there is overlap in terms of administration. This is sometimes referred to herein as “simultaneous” or “concurrent delivery”. In other embodiments, the delivery of one treatment ends before the delivery of the other treatment begins. In some embodiments of either case, the treatment is more effective because of combined administration. For example, the second treatment is more effective, for example, an equivalent effect is seen with less of the second treatment, or the second treatment reduces symptoms to a greater extent, than would be seen if the second treatment were administered in PAT059889-PCT-SEC01 the absence of the first treatment, or the analogous situation is seen with the first treatment. In some embodiments, delivery is such that the reduction in a symptom, or other parameter related to the disorder is greater than what would be observed with one treatment delivered in the absence of the other. The effect of the two treatments can be partially additive, wholly additive, or greater than additive. The delivery can be such that an effect of the first treatment delivered is still detectable when the second is delivered.

[0080] As used herein, a “naive T cell” refers to a T cell that is antigen-inexperienced. In some embodiments, an antigen-inexperienced T cell has encountered its cognate antigen in the thymus but not in the periphery. In some embodiments, naive T cells are precursors of memory cells. In some embodiments, naive T cells express both CD45RA and CCR7, but do not express CD45RO. In some embodiments, naive T cells may be characterized by expression of CD62L, CD27, CCR7, CD45RA, CD28, and CD127, and the absence of CD95 or CD45RO isoform. In some embodiments, naive T cells express CD62L, IL-7 receptor-a, IL-6 receptor, and CD 132, but do not express CD25, CD44, CD69, or CD45RO. In some embodiments, naive T cells express CD45RA, CCR7, and CD62L and do not express CD95 or IL-2 receptor 0. In some embodiments, surface expression levels of markers are assessed using flow cytometry.

[0081] The term “central memory T cells” refers to a subset of T cells that in humans are CD45RO positive and express CCR7. In some embodiments, central memory T cells express CD95. In some embodiments, central memory T cells express IL-2R, IL-7R, and / or IL-15R. In some embodiments, central memory T cells express CD45RO, CD95, IL-2 receptor 0, CCR7, and CD62L. In some embodiments, surface expression levels of markers are assessed using flow cytometry.

[0082] The term “stem memory T cells,” “stem cell memory T cells,” “stem cell-like memory T cells,” “memory stem T cells,” “T memory stem cells,” “T stem cell memory cells,” or “TSCM cells” refers to a subset of memory T cells with stem cell-like ability, for example, the ability to self-renew and / or the multipotent capacity to reconstitute memory and / or effector T cell subsets. In some embodiments, stem memory T cells express CD45RA, CD95, IL-2 receptor 0, CCR7, and CD62L. In some embodiments, surface expression levels of markers are assessed using flow cytometry. In some embodiments, exemplary stem memory T cells are disclosed in Gattinoni et al., Nat Med. 2017 January 06; 23(1): 18-27, herein incorporated by reference in its entirety. PAT059889-PCT-SEC01

[0083] For clarity purposes, unless otherwise noted, classifying a cell or a population of cells as “not expressing,” or having an “absence of’ or being “negative for” a particular marker may not necessarily mean an absolute absence of the marker. The skilled artisan can readily compare the cell against a positive and / or a negative control, and / or set a predetermined threshold, and classify the cell or population of cells as not expressing or being negative for the marker when the cell has an expression level below the predetermined threshold or a population of cells has an overall expression level below the predetermined threshold using conventional detection methods, e.g., using flow cytometry.

[0084] As used herein, the term “GeneSetScore (Up TEM vs. Down TSCM)” of a cell refers to a score that reflects the degree at which the cell shows an effector memory T cell (TEM) phenotype vs. a stem cell memory T cell (TSCM) phenotype. A higher GeneSetScore (Up TEM vs. Down TSCM) indicates an increasing TEM phenotype, whereas a lower GeneSetScore (Up TEM vs. Down TSCM) indicates an increasing TSCM phenotype. In some embodiments, the GeneSetScore (Up TEM vs. Down TSCM) is determined by measuring the expression of one or more genes that are up-regulated in TEM cells and / or down-regulated in TSCM cells, for example, one or more genes selected from the group consisting of MXRA7, CLIC1, NAT13, TBC1D2B, GLCCI1, DUSP10, AP0BEC3D, CACNB3, ANXA2P2, TPRG1, EOMES, MATK, ARHGAP10, ADAM8, MAN1A1, SLFN12L, SH2D2A, EIF2C4, CD58, MY01F, RAB27B, ERN1, NPC1, NBEAL2, AP0BEC3G, SYTL2, SLC4A4, PIK3AP1, PTGDR, MAF, PLEKHA5, ADRB2, PLXND1, GNA01, THBS1, PPP2R2B, CYTH3, KLRF1, FLJ16686, AUTS2, PTPRM, GNLY, and GFPT2. In some embodiments, the GeneSetScore (Up TEM vs. Down TSCM) is determined for each cell using RNA-seq, for example, single-cell RNA-seq (scRNA-seq), for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 39A, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Up TEM vs. Down TSCM) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0085] As used herein, the term “GeneSetScore (Up Treg vs. Down Teff)” of a cell refers to a score that reflects the degree at which the cell shows a regulatory T cell (Treg) phenotype vs. an effector T cell (Teff) phenotype. A higher GeneSetScore (Up Treg vs. Down Teff) indicates an increasing Treg phenotype, whereas a lower GeneSetScore (Up Treg vs. Down Teff) indicates an increasing Teff phenotype. In some embodiments, the GeneSetScore (Up Treg vs. Down Teff) is PAT059889-PCT-SEC01 determined by measuring the expression of one or more genes that are up-regulated in Treg cells and / or down-regulated in Teff cells, for example, one or more genes selected from the group consisting of C12orf75, SELPLG, SWAP70, RGS1, PRR11, SPATS2L, SPATS2L, TSHR, C14orfl45, CASP8, SYT11, ACTN4, ANXA5, GLRX, HLA-DMB, PMCH, RAB11FIP1, IL32, FAM160B1, SHMT2, FRMD4B, CCR3, TNFRSF13B, NTNG2, CLDND1, BARD1, FCER1G, TYMS, ATP1B1, GJB6, FGL2, TK1, SLC2A8, CDKN2A, SKAP2, GPR55, CDCA7, S100A4, GDPD5, PMAIP1, ACOT9, CEP55, SGMS1, ADPRH, AKAP2, HDAC9, IKZF4, CARD17, VAV3, OBFC2A, ITGB1, CIITA, SETD7, HLA-DMA, CCR10, KIAA0101, SLC14A1, PTTG3P, DUSP10, FAM164A, PYHIN1, MY01F, SLC1A4, MYBL2, PTTG1, RRM2, TP53INP1, CCR5, ST8SIA6, TOX, BFSP2, ITPRIPL1, NCAPH, HLA-DPB2, SYT4, NINJ2, FAM46C, CCR4, GBP5, C15orf53, LMCD1, MKI67, NUSAP1, PDE4A, E2F2, CD58, ARHGEF12, LOC100188949, FAS, HLA-DPB1, SELP, WEE1, HLA-DPA1, FCRL1, ICA1, CNTNAP1, OAS1, METTL7A, CCR6, HLA-DRB4, ANXA2P3, STAM, HLA-DQB2, LGALS1, ANXA2, PI 16, DUSP4, LAYN, ANXA2P2, PTPLA, ANXA2P1, ZNF365, LAIR2, LOC541471, RASGRP4, BCAS1, UTS2, MIAT, PRDM1, SEMA3G, FAM129A, HPGD, NCF4, LGALS3, CEACAM4, JAKMIP1, TIGIT, HLA-DRA, IKZF2, HLA-DRB1, FANK1, RTKN2, TRIBI, FCRL3, and FOXP3. In some embodiments, the GeneSetScore (Up Treg vs. Down Teff) is determined using RNA-seq, for example, single-cell RNA-seq (scRNA-seq) , for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 39B, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Up Treg vs. Down Teff) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0086] As used herein, the term “GeneSetScore (Down sternness)” of a cell refers to a score that reflects the degree at which the cell shows a sternness phenotype. A lower GeneSetScore (Down sternness) indicates an increasing sternness phenotype. In some embodiments, the GeneSetScore (Down sternness) is determined by measuring the expression of one or more genes that are upregulated in a differentiating stem cell vs downregulated in a hematopoietic stem cell, for example, one or more genes selected from the group consisting of ACE, BATF, CDK6, CHD2, ERCC2, HOXB4, ME0X1, SFRP1, SP7, SRF, TALI, and XRCC5. In some embodiments, the GeneSetScore (Down sternness) is determined using RNA-seq, for example, single-cell RNA-seq (scRNA-seq) , for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to PAT059889-PCT-SEC01

[0087] FIG. 39C, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Down sternness) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0088] As used herein, the term “GeneSetScore (Up hypoxia)” of a cell refers to a score that reflects the degree at which the cell shows a hypoxia phenotype. A higher GeneSetScore (Up hypoxia) indicates an increasing hypoxia phenotype. In some embodiments, the GeneSetScore (Up hypoxia) is determined by measuring the expression of one or more genes that are up- regulated in cells undergoing hypoxia, for example, one or more genes selected from the group consisting of ABCB1, ACAT1, ADM, AD0RA2B, AK2, AK3, ALDH1A1, ALDH1A3, ALDOA, ALDOC, ANGPT2, ANGPTL4, ANXA1, ANXA2, ANXA5, ARHGAP5, ARSE, ART1, BACE2, BATF3, BCL2L1, BCL2L2, BHLHE40, BHLHE41, BIK, BIRC2, BNIP3, BNIP3L, BPI, BTG1, Cllorf2, C7orf68, CA12, CA9, CALD1, CCNG2, CCT6A, CD99, CDK1, CDKN1A, CDKN1B, CITED2, CLK1, CNOT7, COL4A5, C0L5A1, COL5A2, COL5A3, CP, CTSD, CXCR4, D4S234E, DDIT3, DDIT4, 1-Dec, DKC1, DR1, EDN1, EDN2, EFNA1, EGF, EGR1, EIF4A3, ELF3, ELL2, ENG, EN01, EN03, ENPEP, EPO, ERRFI1, ETS1, F3, FABP5, FGF3, FKBP4, FLT1, FN1, FOS, FTL, GAPDH, GBE1, GLRX, GPI, GPRC5A, HAP1, HBP1, HDAC1, HDAC9, HERC3, HERPUD1, HGF, HIF1A, HK1, HK2, HLA-DQB1, HM0X1, HM0X2, HSPA5, HSPD1, HSPH1, HY0U1, ICAM1, ID2, IFI27, IGF2, IGFBP1, IGFBP2, IGFBP3, IGFBP5, IL6, IL8, INSIGI, IRF6, ITGA5, JUN, KDR, KRT14, KRT18, KRT19, LDHA, LDHB, LEP, LGALS1, LONP1, LOX, LRP1, MAP4, MET, MIF, MMP13, MMP2, MMP7, MPI, MT1L, MTL3P, MUC1, MXI1, NDRG1, NFIL3, NFKB1, NFKB2, NOS1, NOS2, NOS2P1, NOS2P2, NOS3, NR3C1, NR4A1, NT5E, ODC1, P4HA1, P4HA2, PAICS, PDGFB, PDK3, PFKFB1, PFKFB3, PFKFB4, PFKL, PGAM1, PGF, PGK1, PGK2, PGM1, PIM1, PIM2, PKM2, PLAU, PLAUR, PLIN2, PLOD2, PNN, PNP, POLM, PPARA, PPAT, PR0K1, PSMA3, PSMD9, PTGS1, PTGS2, QSOX1, RBPJ, RELA, RI0K3, RNASEL, RPL36A, RRP9, SAT1, SERPINB2, SERPINE1, SGSM2, SIAH2, SIN3A, SIRPA, SLC16A1, SLC16A2, SLC20A1, SLC2A1, SLC2A3, SLC3A2, SLC6A10P, SLC6A16, SLC6A6, SLC6A8, SORL1, SPP1, SRSF6, SSSCA1, STC2, STRA13, SYT7, TBPL1, TCEAL1, TEK, TF, TFF3, TFRC, TGFA, TGFB1, TGFB3, TGFBI, TGM2, TH, THBS1, THBS2, TIMM17A, TNFAIP3, TP53, TPBG, TPD52, TPI1, TXN, TXNIP, UMPS, VEGFA, VEGFB, VEGFC, VIM, VPS11, and XRCC6. In some embodiments, the GeneSetScore (Up hypoxia) is determined using RNA-seq, for example, PAT059889-PCT-SEC01 single-cell RNA-seq (scRNA-seq) , for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 39D, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Up hypoxia) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0089] As used herein, the term “GeneSetScore (Up autophagy)” of a cell refers to a score that reflects the degree at which the cell shows an autophagy phenotype. A higher GeneSetScore (Up autophagy) indicates an increasing autophagy phenotype. In some embodiments, the GeneSetScore (Up autophagy) is determined by measuring the expression of one or more genes that are up-regulated in cells undergoing autophagy, for example, one or more genes selected from the group consisting of ABL1, ACBD5, ACINI, ACTRT1, ADAMTS7, AKR1E2, ALKBH5, ALPK1, AMBRA1, ANXA5, ANXA7, ARSB, ASB2, ATG10, ATG12, ATG13, ATG14, ATG16L1, ATG16L2, ATG2A, ATG2B, ATG3, ATG4A, ATG4B, ATG4C, ATG4D, ATG5, ATG7, ATG9A, ATG9B, ATP13A2, ATP1B1, ATPAF1-AS1, ATPIF1, BECN1, BECN1P1, BLOC1S1, BMP2KL, BNIP1, BNIP3, BOC, Cllorf2, Cl lorf41, C12orf44, C12orf5, C14orfl33, Clorf210, C5, C6orfl06, C7orf59, C7orf68, C8orf59, C9orf72, CA7, CALCB, CALCOCO2, CAPS, CCDC36, CD163L1, CD93, CDC37, CDKN2A, CHAF1B, CHMP2A, CHMP2B, CHMP3, CHMP4A, CHMP4B, CHMP4C, CHMP6, CHST3, CISD2, CLDN7, CLEC16A, CLN3, CLVS1, COX8A, CPA3, CRNKL1, CSPG5, CTSA, CTSB, CTSD, CXCR7, DAP, DKKL1, DNAAF2, DPF3, DRAM1, DRAM2, DYNLL1, DYNLL2, DZANK1, EI24, EIF2S1, EPG5, EPM2A, FABP1, FAM125A, FAM131B, FAM134B, FAM13B, F AMI 76 A, FAM176B, FAM48A, FANCC, FANCF, FANCL, FBXO7, FCGR3B, FGF14, FGF7, FGFBP1, FIS1, FNBP1L, FOXO1, FUNDCI, FUNDC2, FXR2, GAB ARAP, GABARAPL1, GABARAPL2, GABARAPL3, GABRA5, GDF5, GMIP, HAP1, HAPLN1, HBXIP, HCAR1, HDAC6, HGS, HIST1H3A, HIST1H3B, HIST1H3C, HIST1H3D, HIST1H3E, HIST1H3F, HIST1H3G, HIST1H3H, HIST1H3I, HIST1H3J, HK2, HMGB1, HPR, HSF2BP, HSP90AA1, HSPA8, IFI16, IPPK, IRGM, IST1, ITGB4, ITPKC, KCNK3, KCNQ1, KIAA0226, KIAA1324, KRCC1, KRT15, KRT73, LAMP1, LAMP2, LAMTOR1, LAMTOR2, LAMTOR3, LARP1B, LENG9, LGALS8, LIX1, LIX1L, LMCD1, LRRK2, LRSAM1, LSM4, MAP1A, MAP1LC3A, MAP1LC3B, MAP1LC3B2, MAP1LC3C, MAP1S, MAP2K1, MAP3K12, MARK2, MBD5, MDH1, MEX3C, MFN1, MFN2, MLST8, MRPS10, MRPS2, MSTN, MTERFD1, MTMR14, MTMR3, MTOR, MTSS1, MYH11, MYLK, MY0M1, NBR1, PAT059889-PCT-SEC01

[0090] NDUFB9, NEFM, NHLRC1, NME2, NPC1, NR2C2, NRBF2, NTHL1, NUP93, OBSCN, OPTN, P2RX5, PACS2, PARK2, PARK7, PDK1, PDK4, PEX13, PEX3, PFKP, PGK2, PHF23, PHYHIP, PI4K2A, PIK3C3, PIK3CA, PIK3CB, PIK3R4, PINK1, PLEKHM1, PLOD2, PNPO, PPARGC1A, PPY, PRKAA1, PRKAA2, PRKAB1, PRKAB2, PRKAG1, PRKAG2, PRKAG3, PRKD2, PRKG1, PSEN1, PTPN22, RAB12, RAB1A, RAB1B, RAB23, RAB24, RAB33B, RAB39, RAB7A, RB1CC1, RBM18, REEP2, REP15, RFWD3, RGS19, RHEB, RIMS3, RNF185, RNF41, RPS27A, RPTOR, RRAGA, RRAGB, RRAGC, RRAGD, S100A8, S100A9, SCN1A, SERPINB10, SESN2, SFRP4, SH3GLB1, SIRT2, SLC1A3, SLC1A4, SLC22A3, SLC25A19, SLC35B3, SLC35C1, SLC37A4, SLC6A1, SLCO1A2, SMURF1, SNAP29, SNAPIN, SNF8, SNRPB, SNRPB2, SNRPD1, SNRPF, SNTG1, SNX14, SPATAI 8, SQSTM1, SRPX, STAM, STAM2, STAT2, STBD1, STK11, STK32A, STOM, STX12, STX17, SUPT3H, TBC1D17, TBC1D25, TBC1D5, TCIRG1, TEAD4, TECPR1, TECPR2, TFEB, TM9SF1, TMBIM6, TMEM203, TMEM208, TMEM39A, TMEM39B, TMEM59, TMEM74, TMEM93, TNIK, TOLLIP, TOMM20, TOMM22, TOMM40, T0MM5, T0MM6, T0MM7, TOMM70A, TP53INP1, TP53INP2, TRAPPC8, TREM1, TRIM17, TRIM5, TSG101, TXLNA, UBA52, UBB, UBC, UBQLN1, UBQLN2, UBQLN4, ULK1, ULK2, ULK3, USP10, USP13, USP30, UVRAG, VAMP7, VAMP8, VDAC1, VMP1, VPS11, VPS16, VPS18, VPS25, VPS28, VPS33A, VPS33B, VPS36, VPS37A, VPS37B, VPS37C, VPS37D, VPS39, VPS41, VPS4A, VPS4B, VTA1, VTT1A, VTI1B, WDFY3, WDR45, WDR45L, WIPI1, WIPI2, XBP1, YIPF1, ZCCHC17, ZFYVE1, ZKSCAN3, ZNF189, ZNF593, and ZNF681. In some embodiments, the GeneSetScore (Up autophagy) is determined using RNA-seq, for example, single-cell RNA-seq (scRNA-seq), for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 39E, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Up autophagy) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0091] As used herein, the term “GeneSetScore (Up resting vs. Down activated)” of a cell refers to a score that reflects the degree at which the cell shows a resting T cell phenotype vs. an activated T cell phenotype. A higher GeneSetScore (Up resting vs. Down activated) indicates an increasing resting T cell phenotype, whereas a lower GeneSetScore (Up resting vs. Down activated) indicates an increasing activated T cell phenotype. In some embodiments, the GeneSetScore (Up resting vs. Down activated) is determined by measuring the expression of one PAT059889-PCT-SEC01 or more genes that are up-regulated in resting T cells and / or down- regulated in activated T cells, for example, one or more genes selected from the group consisting of ABCA7, ABCF3, ACAP2, AMT, ANKH, ATF7IP2, ATG14, ATP1A1, ATXN7, ATXN7L3B, BCL7A, BEX4, BSDC1, BTG1, BTG2, BTN3A1, Cllorf21, C19orf22, C21orf2, CAMK2G, CARS2, CCNL2, CD248, CD5, CD55, CEP164, CHKB, CLK1, CLK4, CTSL1, DBP, DCUN1D2, DENND1C, DGKD, DLG1, DUSP1, EAPP, ECE1, ECHDC2, ERBB2IP, FAM117A, FAM134B, FAM134C, FAM169A, FAM190B, FAU, FLJ10038, FOXJ2, FOXJ3, FOXL1, FOXO1, FXYD5, FYB, HLA-E, HSPA1L, HYAL2, ICAM2, IFIT5, IFITM1, IKBKB, IQSEC1, IRS4, KIAA0664L3, KIAA0748, KLF3, KLF9, KRT18, LEF1, LINC00342, LIPA, LIPT1, LLGL2, LMBR1L, LPAR2, LTBP3, LYPD3, LZTFL1, MANBA, MAP2K6, MAP3K1, MARCH8, MAU2, MGEA5, MMP8, MPO, MSL1, MSL3, MYH3, MYLIP, NAGPA, NDST2, NISCH, NKTR, NLRP1, NOSIP, NPIP, NUMA1, PAIP2B, PAPD7, PBXIP1, PCIF1, PI4KA, PLCL2, PLEKHA1, PLEKHF2, PNISR, PPFIBP2, PRKCA, PRKCZ, PRKD3, PRMT2, PTP4A3, PXN, RASA2, RASA3, RASGRP2, RBM38, REPIN1, RNF38, RNF44, R0R1, RPL30, RPL32, RPLP1, RPS20, RPS24, RPS27, RPS6, RPS9, RXRA, RYK, SCAND2, SEMA4C, SETD1B, SETD6, SETX, SF3B1, SH2B1, SLC2A4RG, SLC35E2B, SLC46A3, SMAGP, SMARCE1, SMPD1, SNPH, SP140L, SPATA6, SPG7, SREK1IP1, SRSF5, STAT5B, SVIL, SYF2, SYNJ2BP, TAF1C, TBC1D4, TCF20, TECTA, TES, TMEM127, TMEM159, TMEM30B, TMEM66, TMEM8B, TP53TG1, TPCN1, TRIM22, TRIM44, TSC1, TSC22D1, TSC22D3, TSPYL2, TTC9, TTN, UBE2G2, USP33, USP34, VAMP1, VILL, VIPR1, VPS13C, ZBED5, ZBTB25, ZBTB40, ZC3H3, ZFP161, ZFP36L1, ZFP36L2, ZHX2, ZMYM5, ZNF136, ZNF148, ZNF318, ZNF350, ZNF512B, ZNF609, ZNF652, ZNF83, ZNF862, and ZNF91. In some embodiments, the GeneSetScore (Up resting vs. Down activated) is determined using RNA-seq, for example, single-cell RNA-seq (scRNA-seq) , for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 38D, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Up resting vs. Down activated) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0092] As used herein, the term “GeneSetScore (Progressively up in memory differentiation)” of a cell refers to a score that reflects the stage of the cell in memory differentiation. A higher GeneSetScore (Progressively up in memory differentiation) indicates an increasing late memory PAT059889-PCT-SEC01

[0093] T cell phenotype, whereas a lower GeneSetScore (Progressively up in memory differentiation) indicates an increasing early memory T cell phenotype. In some embodiments, the GeneSetScore (Up autophagy) is determined by measuring the expression of one or more genes that are up-regulated during memory differentiation, for example, one or more genes selected from the group consisting of MTCH2, RAB6C, KIAA0195, SETD2, C2orf24, NRD1, GNA13, COP A, SELT, TNIP1, CBFA2T2, LRP10, PRKCI, BRE, ANKS1A, PNPLA6, ARL6IP1, WDFY1, MAPK1, GPR153, SHKBP1, MAP1LC3B2, PIP4K2A, HCN3, GTPBP1, TLN1, C4orf34, KIF3B, TCIRG1, PPP3CA, ATG4D, TYMP, TRAF6, C17orf76, WIPF1, FAM108A1, MYL6, NRM, SPCS2, GGT3P, GALK1, CLIP4, ARL4C, YWHAQ, LPCAT4, ATG2A, IDS, TBC1D5, DMPK, ST6GALNAC6, REEP5, ABHD6, KIAA0247, EMB, TSEN54, SPIRE2, PIWIL4, ZSCAN22, ICAM1, CHD9, LPIN2, SETD8, ZC3H12A, ULBP3, IL15RA, HLA- DQB2, LCP1, CHP, RUNX3, TMEM43, REEP4, MEF2D, ABL1, TMEM39A, PCBP4, PLCD1, CHST12, RASGRP1, Clorf58, Cl lorf63, C6orfl29, FHOD1, DKFZp434F142, PIK3CG, ITPR3, BTG3, C4orf50, CNNM3, IFI16, AK1, CDK2AP1, REL, BCL2L1, MVD, TTC39C, PLEKHA2, FKBP11, EML4, FANCA, CDCA4, FUCA2, MFSD10, TBCD, CAPN2, IQGAP1, CHST11, PIK3R1, MYO5A, KIR2DL3, DLG3, MXD4, RALGDS, S1PR5, WSB2, CCR3, TIP ARP, SP140, CD151, SOX13, KRTAP5-2, NF1, PEA15, PARP8, RNF166, UEVLD, LIMK1, CACNB1, TMX4, SLC6A6, LBA1, SV2A, LLGL2, IRF1, PPP2R5C, CD99, RAPGEF1, PPP4R1, OSBPL7, FOXP4, SLA2, TBC1D2B, ST7, JAZF1, GGA2, PI4K2A, CD68, LPGAT1, STX11, ZAK, FAM160B1, RORA, C8orf80, APOBEC3F, TGFBI, DNAJC1, GPR114, LRP8, CD69, CMIP, NAT13, TGFBI, FLJ00049, ANTXR2, NR4A3, IL12RB1, NTNG2, RDX, MLLT4, GPRIN3, ADCY9, CD300A, SCD5, ABB, PTPN22, LGALS1, SYTL3, BMPR1A, TBK1, PMAIP1, RASGEF1A, GCNT1, GABARAPL1, STOM, CALHM2, ABCA2, PPP1R16B, SYNE2, PAM, C12orf75, CLCF1, MXRA7, APOBEC3C, CLSTN3, ACOT9, HIP1, LAG3, TNFAIP3, DCBLD1, KLF6, CACNB3, RNF19A, RAB27A, FADS3, DLG5, APOBEC3D, TNFRSF1B, ACTN4, TBKBP1, ATXN1, ARAP2, ARHGEF12, FAM53B, MAN1A1, FAM38A, PLXNC1, GRLF1, SRGN, HLA-DRB5, B4GALT5, WIPI1, PTPRJ, SLFN11, DUSP2, ANXA5, AHNAK, NEO1, CLIC1, EIF2C4, MAP3K5, IL2RB, PLEKHG1, MY06, GTDC1, EDARADD, GALM, TARP, ADAM8, MSC, HNRPLL, SYT11, ATP2B4, NHSL2, MATK, ARHGAP18, SLFN12L, SPATS2L, RAB27B, PIK3R3, TP53INP1, MBOAT1, GYG1, KATNAL1, FAM46C, ZC3HAV1L, ANXA2P2, CTNNA1, NPC1, C3AR1, PAT059889-PCT-SEC01

[0094] CRIM1, SH2D2A, ERN1, YPEL1, TBX21, SLC1A4, FASLG, PHACTR2, GALNT3, ADRB2, PIK3AP1, TLR3, PLEKHA5, DUSP10, GNAO1, PTGDR, FRMD4B, ANXA2, EOMES, CADM1, MAF, TPRG1, NBEAL2, PPP2R2B, PELO, SLC4A4, KLRF1, FOSL2, RGS2, TGFBR3, PRF1, MYO1F, GAB3, C17orf66, MICAL2, CYTH3, TOX, HLA-DRA, SYNE1, WEE1, PYHIN1, F2R, PLD1, THBS1, CD58, FAS, NETO2, CXCR6, ST6GALNAC2, DUSP4, AUTS2, Clorf21, KLRG1, TNIP3, GZMA, PRR5L, PRDM1, ST8SIA6, PLXND1, PTPRM, GFPT2, MYBL1, SLAMF7, FLJ16686, GNLY, ZEB2, CST7, IL18RAP, CCL5, KLRD1, and KLRB1. In some embodiments, the GeneSetScore (Progressively up in memory differentiation) is determined using RNA-seq, for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 40B, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Progressively up in memory differentiation) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0095] As used herein, the term “GeneSetScore (Up TEM vs. Down TN)” of a cell refers to a score that reflects the degree at which the cell shows an effector memory T cell (TEM) phenotype vs. a naive T cell (TN) phenotype. A higher GeneSetScore (Up TEM vs. Down TN) indicates an increasing TEM phenotype, whereas a lower GeneSetScore (Up TEM vs. Down TN) indicates an increasing TN phenotype. In some embodiments, the GeneSetScore (Up TEM vs. Down TN) is determined by measuring the expression of one or more genes that are up-regulated in TEM cells and / or down-regulated in TN cells, for example, one or more genes selected from the group consisting of MY05A, MXD4, STK3, S1PR5, GLCCI1, CCR3, SOX13, KRTAP5-2, PEA15, PARP8, RNF166, UEVLD, LIMK1, SLC6A6, SV2A, KPNA2, OSBPL7, ST7, GGA2, PI4K2A, CD68, ZAK, RORA, TGFBI, DNAJC1, JOSD1, ZFYVE28, LRP8, OSBPL3, CMIP, NAT13, TGFBI, ANTXR2, NR4A3, RDX, ADCY9, CHN1, CD300A, SCD5, PTPN22, LGALS1, RASGEF1A, GCNT1, GLUL, ABCA2, CLDND1, PAM, CLCF1, MXRA7, CLSTN3, ACOT9, METRNL, BMPR1A, LRIG1, APOBEC3G, CACNB3, RNF19A, RAB27A, FADS3, ACTN4, TBKBP1, FAM53B, MAN1A1, FAM38A, GRLF1, B4GALT5, WIPI1, DUSP2, ANXA5, AHNAK, CLIC1, MAP3K5, ST8SIA1, TARP, ADAM8, MATK, SLFN12L, PIK3R3, FAM46C, ANXA2P2, CTNNA1, NPC1, SH2D2A, ERN1, YPEL1, TBX21, STOM, PHACTR2, GBP5, ADRB2, PIK3AP1, DUSP10, PTGDR, EOMES, MAF, TPRG1, NBEAL2, NCAPH, SLC4A4, FOSL2, RGS2, TGFBR3, MYO1F, C17orf66, CYTH3, WEE1, PYHIN1, F2R, THBS1, CD58, AUTS2, FAM129A, TNIP3, GZMA, PRR5L, PRDM1, PLXND1, PTPRM, PAT059889-PCT-SEC01

[0096] GFPT2, MYBL1, SLAMF7, ZEB2, CST7, CCL5, GZMK, and KLRB1. In some embodiments, the GeneSetScore (Up TEM vs. Down TN) is determined using RNA-seq, for example, singlecell RNA-seq (scRNA-seq), for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 40C, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Up TEM vs. Down TN) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0097] In the context of GeneSetScore values (e.g., median GeneSetScore values), when a positive GeneSetScore is reduced by 100%, the value becomes 0. When a negative GeneSetScore is increased by 100%, the value becomes 0. For example, as disclosed in WO / 2020 / 047452, the median GeneSetScore of the Dayl sample is -0.084; the median GeneSetScore of the Day9 sample is 0.035; and the median GeneSetScore of the input sample is -0.1. In WO / 2020 / 047452 in FIG. 39A, increasing the median GeneSetScore of the input sample by 100% leads to a GeneSetScore value of 0; and increasing the median GeneSetScore of the input sample by 200% leads to a GeneSetScore value of 0.1. In WO / 2020 / 047452 in FIG. 39A, decreasing the median GeneSetScore of the Day9 sample by 100% leads to a GeneSetScore value of 0; and decreasing the median GeneSetScore of the Day9 sample by 200% leads to a GeneSetScore value of -0.035.

[0098] Various embodiments of the compositions and methods herein are described in further detail below. Additional definitions are set out throughout the specification.

[0099] CD19 CAR-Expressing Cells

[0100] The present disclosure provides CAR-expressing cell compositions and their use in medicaments or methods for treating, among other diseases, myasthenia gravis (MG), e.g., early- onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, involving cells or tissues which express an antigen as described herein. In some embodiments, provided herein are pharmaceutical compositions comprising a CAR-expressing cell, for example, a plurality of CD 19 CAR-expressing cells, in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents or excipients. In some embodiments, the CD 19 CAR-expressing cells is rapcabtag ene autoleucel. PAT059889-PCT-SEC01

[0101] In some embodiments, the antigen-binding domain of the CD 19 CAR has the same or a similar binding specificity as the FMC63 scFv fragment described in Nicholson et al. Mol. Immun. 34 (16-17): 1157-1165 (1997). In some embodiments, the antigen-binding domain of the CD19 CAR includes the scFv fragment described in Nicholson et al. Mol. Immun. 34 (16- 17): 1157-1165 (1997). In some embodiments, the CD19 CAR includes an antigen-binding domain (for example, a humanized antigen-binding domain) according to Table 3 of WO2014 / 153270, incorporated herein by reference. In some embodiments, the CD19 CAR comprises an amino acid sequence provided as SEQ ID NO: 12 in PCT publication WO2012 / 079000, incorporated herein by reference.

[0102] In some embodiments, the population of CAR T cells that specifically bind to CD 19 comprises rapcabtagene autoleucel. The rapcabtagene autoleucel is made using autologous T cells obtained from peripheral blood mononuclear cells from a subject (e.g., from a subject having an neuroimmune disease or disorder) by leukapheresis and subsequently transduced with a self-inactivating, non-replicating lentiviral vector encoding a T cell chimeric antigen receptor targeting CD 19. The expressed transgene comprises a CD8a leader sequence, a murine anti- CD19 single chain variable fragment (scFv) derived from the mouse hybridoma FMC63, a CD8a hinge and transmembrane region, and a 4- IBB (CD 137) and CD3(j (TCRQ signaling domain, and is under control of the elongation factor 1 alpha (EFla) promoter. The construct is flanked by 5' and 3' long terminal repeats (LTRs) and also contains a \| / packaging signal, a Rev response element (RRE), a central polypurine tract (cPPT) sequence, and an optimized Woodchuck hepatitis virus posttranscriptional regulatory element (WPRE). The leukapheresis material is enriched for CD4 / CD8 T cells by positive immunoselection, activated by CD3 and CD28 agonists and transduced with the vector. Without further cell propagation, the T cells are washed, formulated for infusion, and cryopreserved. Rapcabtagene autoleucel is composed of >80% T cells and <1% B cells, with a mixture of transgene positive (>3.4%) and negative T cells. The CD4+ and CD8+ naive T cell subsets (CD45RA+CCR7+) present in the leukapheresis material are largely retained. In some embodiments, the rapcabtagene autoleucel is made by the ARM manufacturing process described herein.

[0103] In some embodiments, the CD19 CAR-expressing cell population has:

[0104] (a) about the same percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, as compared to the percentage of naive cells, for example, PAT059889-PCT-SEC01 naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;

[0105] (b) a change within about 5% to about 10% of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, for example, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;

[0106] (c) an increased percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, for example, increased by at least 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, or 3-fold, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;

[0107] (d) about the same percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0108] (e) a change within about 5% to about 10% of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0109] (f) a decreased percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, for example, decreased by at least 20, 25, 30, 35, 40, 45, or 50%, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0110] (g) about the same percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0111] (h) a change within about 5% to about 10% of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of PAT059889-PCT-SEC01 stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR; or

[0112] (i) an increased percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR.

[0113] In some embodiments, the CD 19 CAR-expressing population of cells made via the ARM process described herein shows a higher percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells (for example, at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50% higher), compared with cells made by an otherwise similar method which lasts, for example, more than 26 hours (for example, which lasts more than 5, 6, 7, 8, 9, 10, 11, or 12 days) or which involves expanding the population of cells in vitro for, for example, more than 3 days (for example, expanding the population of cells in vitro for 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days).

[0114] In some embodiments, the CD 19 CAR-expressing population of cells made via the ARM process described herein shows a lower percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells (for example, at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50% lower), compared with cells made by an otherwise similar method which lasts, for example, more than 26 hours (for example, which lasts more than 5, 6, 7, 8, 9, 10, 11, or 12 days) or which involves expanding the population of cells in vitro for, for example, more than 3 days (for example, expanding the population of cells in vitro for 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days).

[0115] In some embodiments, the CD 19 CAR-expressing population of cells made via the ARM process described herein after being administered in vivo, persists longer or expands at a higher level (for example, at least 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90% higher), compared with cells made by an otherwise similar method which lasts, for example, more than 26 hours (for example, which lasts more than 5, 6, 7, 8, 9, 10, 11, or 12 days) or which involves expanding the population of cells in vitro for, for example, more than 3 days (for example, expanding the population of cells in vitro for 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days).

[0116] Any known CD 19 CAR, for example, the CD 19 antigen-binding domain of any known CD 19 CAR, in the art can be used in accordance with the present disclosure. Examples include PAT059889-PCT-SEC01 tisagenlecleucel, axicabtagene ciloleucel, brexucabtagene autoleucel, lisocabtagene maraleucel, relmacabtagene autoleucel, CC-97540, AT101, CABA-201, KYV-101, IMPT-514, or MB- CART19.1. Further exemplary CD 19 CARs include CD 19 CARs described herein or an antiCD F CAR described in Xu et al. Blood 123.24(2014):3750-9; Kochenderfer et al. Blood 122.25(2013):4129-39, Cruz et al. Blood 122.17(2013):2965-73, NCT00586391, NCT01087294, NCT02456350, NCT00840853, NCT02659943, NCT02650999, NCT02640209, NCT01747486, NCT02546739, NCT02656147, NCT02772198, NCT00709033, NCT02081937, NCT00924326, NCT02735083, NCT02794246, NCT02746952, NCT01593696, NCT02134262, NCT01853631, NCT02443831, NCT02277522, NCT02348216, NCT02614066, NCT02030834, NCT02624258, NCT02625480, NCT02030847, NCT02644655, NCT02349698, NCT02813837, NCT02050347, NCT01683279, NCT02529813, NCT02537977, NCT02799550, NCT02672501, NCT02819583, NCT02028455, NCT01840566, NCT01318317, NCT01864889, NCT02706405, NCT01475058, NCT01430390, NCT02146924, NCT02051257, NCT02431988, NCT01815749, NCT02153580, NCT01865617, NCT02208362, NCT02685670, NCT02535364, NCT02631044, NCT02728882, NCT02735291, NCT01860937, NCT02822326, NCT02737085, NCT02465983, NCT02132624, NCT02782351, NCT01493453, NCT02652910, NCT02247609, NCT01029366, NCT01626495, NCT02721407, NCT01044069, NCT00422383, NCT01680991, NCT02794961, or NCT02456207, NCT05338931, NCT05869955, NCT06056921, NCT06106906, NCT06121297, NCT06152172, NCT06189157, NCT05459870, NCT06153095, each of which is incorporated herein by reference in its entirety.

[0117] Methods of Treating

[0118] The present application discloses methods of treating method of treating a subject having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, the method comprising administering to the subject a population of cells engineered to express a CD 19 CAR (“a population of CD19 CAR-expressing cells”). In some embodiments, the method further comprises identifying the subject as a responder or a non- responder, and / or one who has achieved remission, based on a measure of one or more of the parameters of efficacy disclosed herein. In some embodiments, the measure of one or more of the parameters disclosed herein is PAT059889-PCT-SEC01 obtained from a sample acquired from the subject. In some embodiments of any of the methods disclosed herein, the subject is evaluated prior to receiving, during, or after receiving, the CD 19 CAR-expressing cell therapy. In some embodiments, the CD 19 CAR-expressing cell therapy is rapcabtagene autoleucel.

[0119] In some embodiments, the identification of a subject as a responder, non- responder and / or one who has achieved remission, following administration of the CD 19 CAR-expressing cells according to the methods described herein can be determined by evaluating the subject according to clinical criteria. Efficacy may be evaluated, for example, using the Myasthenia Gravis-Activities of Daily Living (MG-ADL), Quantitative Myasthenia Gravis Scale (QMG), MGFA Post-Intervention Status (PIS), Myasthenia gravis-Quality of Life 15-reviseed (MG-QoL 15r), or Short Form Health Survey (SF-36). The above criteria are explained in further detail below.

[0120] The Myasthenia Gravis Activities of Living (MG-ADL) scale is an 8-item patient- reported scale that measures myasthenia gravis (MG) symptoms and functional status, to be administered by physician or trained study evaluator. The MG-ADL is an outcome measure assessing MG symptoms and functional activities related to activities of daily living (Wolfe et al 1999). Each of the items is scored from 0 (normal) to 3 (most severe), providing a total MG- ADL score ranging from 0 to 24, where higher scores indicate greater severity of symptoms. The MG-ADL is composed of items related to patients’ assessment of functional disability secondary to ocular (two items), bulbar (three items), respiratory (one item), and gross motor or limb impairment (two items). Items are linearly scored and not weighted, with each item ranging from 0 to 3 for a total score range of 0 to 24. The MG-ADL is easy to administer, is quick to complete (<10 min), and can be used in routine clinical practice or in clinical trials. Change in this scale has been used in pivotal trials as primary outcome parameter to assess MG disease severity. A two-point reduction in MG-ADL total score optimally indicates improvement for patients with MG based on a receiver operator characteristic curve approach. This threshold of a two-point difference is recognized as a definition of responder with clinically important improvement.

[0121] In some embodiments, a subject is identified as showing improvement as measured by a decrease in the MG-ADL score from baseline following administration of the CD 19 CAR- expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, the baseline MG-ADL score of the patient is the MG-ADL score PAT059889-PCT-SEC01 measured prior to the first administration of the CD 19 CAR-expressing cells. In some embodiments, the baseline MG-ADL score of the patient is the first MG-ADL score measured after administration of the CD 19 CAR-expressing cells.

[0122] In some embodiments, the MG-ADL score does not worsen. In some embodiments, the MG-ADL score is lowered by at least 2 points. In some embodiments, the MG-ADL score is lowered by at least 3 points. In some embodiments, the EDSS score is lowered by at least 4 points. In some embodiments, the MG-ADL score is lowered by at least 5 points. In some embodiments, the MG-ADL score is lowered by at least 6 points. In some embodiments, the MG-ADL score is lowered by at least 7 points. In some embodiments, the MG-ADL score is lowered by at least 8 points. In some embodiments, the MG-ADL score is lowered by at least 9 points. In some embodiments, the MG-ADL score is lowered by at least 10 points. In some embodiments, the MG-ADL score is lowered by at least 11 points. In some embodiments, the MG-ADL score is lowered by at least 12 points. In some embodiments, the MG-ADL score is lowered by at least 13 points. In some embodiments, the MG-ADL score is lowered by at least 14 points. In some embodiments, the MG-ADL score is lowered by at least 15 points. In some embodiments, the MG-ADL score is lowered by at least 16 points. In some embodiments, the MG-ADL score is lowered by at least 17 points. In some embodiments, the MG-ADL score is lowered by at least 18 points. In some embodiments, the MG-ADL score is lowered by at least 19 points. In some embodiments, the MG-ADL score is lowered by at least 20 points. In some embodiments, the MG-ADL score is lowered by at least 21 points. In some embodiments, the MG-ADL score is lowered by at least 22 points. In some embodiments, the MG-ADL score is lowered by at least 23 points. In some embodiments, the MG-ADL score is lowered by at least 24 points.

[0123] In some embodiments, a subject is identified as showing improvement as measured by a lowering of the MG-ADL score from baseline following administration of the CD 19 CAR- expressing cells (e.g., rapcabtag ene autoleucel) to the subject according to the methods described herein. In some embodiments, the lowering of the MG-ADL score is achieved within 3 months of administration. In some embodiments, lowering of the MG-ADL score is achieved within 6 months of administration. In some embodiments, the lowering of the MG-ADL score is achieved within 9 months of administration. In some embodiments, lowering of the MG-ADL score is achieved within 12 months of administration. In some embodiments, the lowering of the MG- PAT059889-PCT-SEC01

[0124] ADL score is achieved within 18 months of administration. In some embodiments, lowering of the MG- ADL score is achieved within 24 months of administration.

[0125] In some embodiments, the lowering of the MG- ADL score from baseline achieved by a subject following administration of the CD 19 CAR-expressing cells (e.g., rapcabtag ene autoleucel) is sustained for 3 months or more. In some embodiments, lowering of the MG- ADL score is sustained for 6 months or more. In some embodiments lowering of the MG- ADL score is sustained for 9 months or more. In some embodiments, the lowering of the MG- ADL score is sustained for 12 months or more. In some embodiments, the lowering of the MG- ADL score is sustained for 18 months or more. In some embodiments, the lowering of the MG- ADL score is sustained for 24 months or more. In some embodiments, the lowering of the MG- ADL score is sustained indefinitely.

[0126] The Quantitative Myasthenia Gravis Scale (“QMG”) score is a standardized quantitative strength scoring system developed specifically for MG. The QMG has been validated and has been used in several previous MG trials. The QMG is a 13 -item direct physician assessment scoring system that quantifies disease severity based on impairments of body functions and structures (Barohn et al 1998). Each item is quantitatively assessed and scored from 0 to 3 (where 3 represents the most severe), providing a total QMG score ranging from 0 to 39. The QMG is composed of the following items: ocular (two items), facial (one item), bulbar (two items), gross motor (six items), axial (one item), and respiratory (one item).

[0127] In some embodiments, a subject is identified as showing improvement as measured by a lowering of the QMG score from baseline following administration of the CD 19 CAR- expressing cells (e.g., rapcabtag ene autoleucel) to the subject according to the methods described herein. In some embodiments, the baseline QMG score of the patient is the QMG score measured prior to the first administration of the CD 19 CAR-expressing cells. In some embodiments, the baseline QMG score of the patient is the first QMG score measured after administration of the CD 19 CAR-expressing cells.

[0128] In some embodiments, the QMG score is lowered by 2 or more points. In some embodiments, the QMG score is lowered by 3 or more points. In some embodiments, the QMG score is lowered by 4 or more points. In some embodiments, the QMG score is lowered by 5 or more points. In some embodiments, the QMG score is lowered by 6 or more points. In some embodiments, the QMG score is lowered by 7 or more points. In some embodiments, the QMG PAT059889-PCT-SEC01 score is lowered by 8 or more points. In some embodiments, the QMG score is lowered by 9 or more points. In some embodiments, the QMG score is lowered by 10 or more points. In some embodiments, the QMG score is lowered by 11 or more points. In some embodiments, the QMG score is lowered by 12 or more points. In some embodiments, the QMG score is lowered by 13 or more points. In some embodiments, the QMG score is lowered by 14 or more points. In some embodiments, the QMG score is lowered by 15 or more points. In some embodiments, the QMG score is lowered by 16 or more points. In some embodiments, the QMG score is lowered by 17 or more points. In some embodiments, the QMG score is lowered by 18 or more points. In some embodiments, the QMG score is lowered by 19 or more points. In some embodiments, the QMG score is lowered by 20 or more points. In some embodiments, the QMG score is lowered by 21 or more points. In some embodiments, the QMG score is lowered by 22 or more points. In some embodiments, the QMG score is lowered by 23 or more points. In some embodiments, the QMG score is lowered by 24 or more points. In some embodiments, the QMG score is lowered by 25 or more points. In some embodiments, the QMG score is lowered by 26 or more points. In some embodiments, the QMG score is lowered by 27 or more points. In some embodiments, the QMG score is lowered by 28 or more points. In some embodiments, the QMG score is lowered by 29 or more points. In some embodiments, the QMG score is lowered by 30 or more points. In some embodiments, the QMG score is lowered by 31 or more points. In some embodiments, the QMG score is lowered by 32 or more points. In some embodiments, the QMG score is lowered by 33 or more points. In some embodiments, the QMG score is lowered by 34 or more points. In some embodiments, the QMG score is lowered by 35 or more points. In some embodiments, the QMG score is lowered by 36 or more points. In some embodiments, the QMG score is lowered by 37 or more points. In some embodiments, the QMG score is lowered by 38 or more points. In some embodiments, the QMG score is lowered by 39 or more points.

[0129] In some embodiments, a subject is identified as showing improvement as measured by a lowering of the QMG score from baseline following administration of the CD 19 CAR- expressing cells (e.g., rapcabtag ene autoleucel) to the subject according to the methods described herein. In some embodiments, the lowering of the QMG score is achieved within 3 months of administration. In some embodiments, lowering of the QMG score is achieved within 6 months of administration. In some embodiments, the lowering of the QMG score is achieved within 9 months of administration. In some embodiments, lowering of the QMG score is achieved within PAT059889-PCT-SEC01

[0130] 12 months of administration. In some embodiments, the lowering of the QMG score is achieved within 18 months of administration. In some embodiments, lowering of the QMG score is achieved within 24 months of administration.

[0131] In some embodiments, the lowering of the QMG score from baseline achieved by a subject following administration of the CD 19 CAR-expressing cells (e.g., rapcabtag ene autoleucel) is sustained for 3 months or more. In some embodiments, lowering of the QMG score is sustained for 6 months or more. In some embodiments lowering of the QMG score is sustained for 9 months or more. In some embodiments, the lowering of the QMG score is sustained for 12 months or more. In some embodiments, the lowering of the QMG score is sustained for 18 months or more. In some embodiments, the lowering of the QMG score is sustained for 24 months or more. In some embodiments, the lowering of the QMG score is sustained indefinitely.

[0132] The Myasthenia Gravis Foundation of America Post-Intervention Status (MGFA-PIS) is a physician-assessed determination of the overall clinical state of an MG patient at any time after initiation of treatment for MG. (Jaretzki et al 2000). It can be used to evaluate changes in a patient's condition following treatment, including improvement, worsening, or no change of clinical manifestations from pretreatment. It provides the physician’s global assessment of the patient’s clinical status according to the following criteria: PAT059889-PCT-SEC01

[0133] In some embodiments, a subject is identified as having an improved status relative to baseline according to the MGFA-PIS following administration of the CD 19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, the baseline MGFA-PIS status is determined prior to the first administration of the CD 19 CAR-expressing cells. In some embodiments, the baseline MGFA-PIS status is the first evaluation of MGFA-PIS status after administration of the CD 19 CAR-expressing cells. PAT059889-PCT-SEC01

[0134] In some embodiments, a subject is identified as improved according to the MGFA-PIS by achieving complete stable remission following administration of the CD 19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, a subject is identified as improved according to the MGFA-PIS by achieving pharmacologic remission following administration of the CD 19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, a subject is identified as improved according to the MGFA-PIS by achieving minimal manifestations following administration of the CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein.

[0135] In some embodiments, a subject is identified as improved according to the MGFA-PIS by achieving MM-3 following administration of the CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, a subject is identified as improved according to the MGFA-PIS by achieving MM-2 following administration of the CD 19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, a subject is identified as improved according to the MGFA-PIS by achieving MM-1 following administration of the CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, a subject is identified as improved according to the MGFA-PIS by achieving MM-0 following administration of the CD 19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein.

[0136] In some embodiments, a subject is identified as having an improved status (complete stable remission, pharmacologic remission, or minimal manifestations, e.g., MM-3, MM-2, MM- 1, or MM-0) according to the MGFA-PIS following administration of the CD 19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, the improved status is achieved within 3 months of administration. In some embodiments, the improved status is achieved within 6 months of administration. In some embodiments, the improved status is achieved within 9 months of administration. In some embodiments, the improved status is achieved within 12 months of administration. In some embodiments, the improved status is achieved within 18 months of administration. In some embodiments, the improved status is achieved within 24 months of administration. PAT059889-PCT-SEC01

[0137] In some embodiments, the improved status (complete stable remission, pharmacologic remission, or minimal manifestations, e.g., MM-3, MM-2, MM-1, or MM-0) according to the MGFA-PIS achieved by a subject following administration of the CD 19 CAR-expressing cells (e.g., rapcabtagene autoleucel) is sustained for 3 months or more. In some embodiments, the improved status is sustained for 6 months or more. In some embodiments, the improved status is sustained for 9 months or more. In some embodiments, the improved status is sustained for 12 months or more. In some embodiments, the improved status is sustained for 18 months or more. In some embodiments, the improved status is sustained for 24 months or more. In some embodiments, the improved status is sustained indefinitely.

[0138] The 36-Item Short Form Survey (SF-36) is an outcome measure instrument that is often used, well-researched, self-reported measure of health. The SF-36 yields scores for eight scales: Physical Functioning (PF), Role Limitations due to Physical Health (RP), Bodily Pain (BP), General Health (GH), Vitality (VT), Social Functioning (SF), Role Limitations due to Emotional Health (RE), and Mental Health (MH). These scale scores can be combined to produce two component summary scores: the Physical Component Summary (PCS) and the Mental Component Summary (MCS), whose calculation produces a T-score ranging from 0 to 100, with a mean score of 50 and SD of 10, representing the reference score for the US general population. Higher scores indicate better health.

[0139] In some embodiments, a subject is identified as showing improvement as measured by an increase of the SF-36 score from baseline following administration of the CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, the baseline SF-36 score of the patient is the SF-36 score measured prior to the first administration of the CD 19 CAR-expressing cells. In some embodiments, the baseline SF-36 score of the patient is the first SF-36 score measured after administration of the CD 19 CAR-expressing cells.

[0140] In some embodiments, the SF-36 score does not worsen. In some embodiments, the SF- 36 score is increased by 5 or more points. In some embodiments, the SF-36 score is increased by 10 or more points. In some embodiments, the SF-36 score is increased by 15 or more points. In some embodiments, the SF-36 score is increased by 20 or more points. In some embodiments, the SF-36 score is increased by 25 or more points. In some embodiments, the SF-36 score is increased by 30 or more points. In some embodiments, the SF-36 score is increased by 35 or PAT059889-PCT-SEC01 more points. In some embodiments, the SF-36 score is increased by 40 or more points. In some embodiments, the SF-36 score is increased by 45 or more points. In some embodiments, the SF- 36 score is increased by 50 or more points.

[0141] In some embodiments, a subject is identified as showing improvement as measured by an increase of the SF-36 score from baseline following administration of the CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, the increase of the SF-36 score is achieved within 3 months of administration. In some embodiments, increase of the SF-36 score is achieved within 6 months of administration. In some embodiments, the increase of the SF-36 score is achieved within 9 months of administration. In some embodiments, the increase of the SF-36 score is achieved within 12 months of administration. In some embodiments, the increase of the SF-36 score is achieved within 18 months of administration. In some embodiments, the increase of the SF- 36score is achieved within 24 months of administration.

[0142] In some embodiments, the increase of the SF-36 score from baseline achieved by a subject following administration of the CD 19 CAR-expressing cells (e.g., rapcabtagene autoleucel) is sustained for 3 months or more. In some embodiments, increase of the SF-36 score is sustained for 6 months or more. In some embodiments, the increase of the SF-36 score is sustained for 9 months or more. In some embodiments, the increase of the SF-36 score is sustained for 12 months or more. In some embodiments, the increase of the SF-36 score is sustained for 18 months or more. In some embodiments, the increase of the SF-36 score is sustained for 24 months or more. In some embodiments, the increase of the SF-36 score is sustained indefinitely.

[0143] The Myasthenia Gravis-Quality of Life 15 revised (MG-QoL-15r) a 15-question scale designed to assess aspects of life related to myasthenia gravis. There is an appropriate correlation between the MG-QOL 15r and other MG-specific scales (MGC, MG-ADL, and QMG score). The MG-QoL15r allows clinicians to estimate a patient’s quality of life relevant to MG (Burns et al 2011). Items on the MG-QoL15r relate to physical, social, and psychological components and are scored from 0 (not at all) to 4 (quite a bit). The cumulative scores range from 0 to 60, with higher scores representing worse quality of life.

[0144] In some embodiments, a subject is identified as showing improvement as measured by a decrease of in the MG-QoL- 15r score from baseline following administration of the CD 19 CAR- PAT059889-PCT-SEC01 expressing cells (e.g., rapcabtag ene autoleucel) to the subject according to the methods described herein. In some embodiments, the baseline MG-QoL-15r score of the patient is the MG-QoL-15r score measured prior to the first administration of the CD 19 CAR-expressing cells. In some embodiments, the baseline MG-QoL-15r score of the patient is the first MG-QoL-15r score measured after administration of the CD 19 CAR-expressing cells.

[0145] In some embodiments, the MG-QoL-15r score is decreased by at least 5 points. In some embodiments, the MG-QoL-15r score is decreased by at least 10 points. In some embodiments, the MG-QoL-15r score is decreased by at least 15 points. In some embodiments, the MG-QoL- 15r score is decreased by at least 20 points. In some embodiments, the MG-QoL-15r score is decreased by at least 25 points. In some embodiments, the MG-QoL-15r score is decreased by at least 30 points. In some embodiments, the MG-QoL-15r score is decreased by at least 35 points. In some embodiments, the MG-QoL-15r score is decreased by at least 40 points. In some embodiments, the MG-QoL-15r score is decreased by at least 45 points. In some embodiments, the MG-QoL-15r score is decreased by at least 50 points. In some embodiments, the MG-QoL- 15r score is decreased by at least 55 points. In some embodiments, the MG-QoL-15r score is decreased by at least 60 points.

[0146] In some embodiments, a subject is identified as showing improvement as measured by a decrease in the MG-QoL-15r score from baseline following administration of the CD 19 CAR- expressing cells (e.g., rapcabtag ene autoleucel) to the subject according to the methods described herein. In some embodiments, the decrease in the MG-QoL-15r score is achieved within 3 months of administration. In some embodiments, decrease in the MG-QoL-15r score is achieved within 6 months of administration. In some embodiments, the decrease in the MG-QoL- 15r score is achieved within 9 months of administration. In some embodiments, the decrease in the MG- QoL-15r score is achieved within 12 months of administration. In some embodiments, the decrease in the MG-QoL- 15r score is achieved within 18 months of administration. In some embodiments, the decrease in the MG-QoL- 15r score is achieved within 24 months of administration.

[0147] In some embodiments, the reduction in the MG-QoL- 15r score from baseline achieved by a subject following administration of the CD 19 CAR-expressing cells (e.g., rapcabtagene autoleucel) is sustained for 3 months or more. In some embodiments, the reduction in the MG- QoL-15r score is sustained for 6 months or more. In some embodiments, the reduction in the PAT059889-PCT-SEC01

[0148] MG-QoL-15r score is sustained for 9 months or more. In some embodiments, the reduction in the MG-QoL-15r score is sustained for 12 months or more. In some embodiments, the reduction in the MG-QoL-15r score is sustained for 18 months or more. In some embodiments, the reduction in the MG-QoL-15r score is sustained for 24 months or more. In some embodiments, the reduction in the MG-QoL-15r score is sustained indefinitely.

[0149] In some embodiments, a subject is identified as showing improvement as measured by a decrease in the level of autoantibodies, e.g., AChR, LRP4, MuSK antibodies, from baseline following administration of the CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, the baseline autoantibody level of the patient is the autoantibody level measured prior to the first administration of the CD 19 CAR-expressing cells. In some embodiments, the baseline autoantibody level of the patient is the first autoantibody level measured after administration of the CD 19 CAR-expressing cells.

[0150] In some embodiments, a subject is identified as showing improvement as measured by a decrease in the autoantibody level, e.g., AChR, LRP4, MuSK antibodies, from baseline following administration of the CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, the decrease in the autoantibody level is achieved within 3 months of administration. In some embodiments, decrease in the autoantibody level is achieved within 6 months of administration. In some embodiments, the decrease in the autoantibody level is achieved within 9 months of administration. In some embodiments, the decrease in the autoantibody level is achieved within 12 months of administration. In some embodiments, the decrease in the autoantibody level is achieved within 18 months of administration. In some embodiments, the decrease in the autoantibody level is achieved within 24 months of administration.

[0151] In some embodiments, the reduction in the autoantibody level, e.g., AChR, LRP4, MuSK antibodies, from baseline achieved by a subject following administration of the CD 19 CAR- expressing cells (e.g., rapcabtagene autoleucel) is sustained for 3 months or more. In some embodiments, the reduction in the autoantibody level is sustained for 6 months or more. In some embodiments, the reduction in the autoantibody level is sustained for 9 months or more. In some embodiments, the reduction in the autoantibody level is sustained for 12 months or more. In some embodiments, the reduction in the autoantibody level is sustained for 18 months or more. PAT059889-PCT-SEC01

[0152] In some embodiments, the reduction in the autoantibody level is sustained for 24 months or more. In some embodiments, the reduction in the autoantibody level is sustained indefinitely.

[0153] Dosage regimen

[0154] In some embodiments, a dose of viable CD 19 CAR-expressing cells comprises about 0.5 x 106viable CAR-expressing cells to about 1.25 x 109viable CAR-expressing cells (for example, 0.5 x 106viable CAR-expressing cells to 1.25 x 109viable CAR-expressing cells). In some embodiments, a dose of viable CAR-expressing cells comprises about 1 x 106, about 2.5 x 106, about 5 x 106, about 1.25 x 107, about 2.5 x 107, about 5 x 107, about 5.75 x 107, or about 8 x 107viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 0.5 x 106to 90 x 106viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 5 x 106viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 2.5 x 106to 2.5 x 108viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 1.25 x 107viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 1.25 x 107to 1.25 x 109viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 1.25 x 108viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 2.5 x 106to 2.5 x 108viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 1 x 107or 5 x 107viable CAR-expressing cells. In some embodiments, the viable CD 19 CAR-expressing cells are rapcabtagene autoleucel.

[0155] In some embodiments, a dose of CAR-expressing cells comprises about 1 x 106, 1.1 x 106, 2 x 106, 3.6 x 106, 5 x 106, 1 x 107, 1.8 x 107, 2 x 107, 5 x 107, 1 x 108, 2 x 108, or 5 x 108cells / kg. In some embodiments, a dose of CAR-expressing cells comprises at least about 1 x 106, 1.1 x 106, 2 x 106, 3.6 x 106, 5 x 106, 1 x 107, 1.8 x 107, 2 x 107, 5 x 107, 1 x 108, 2 x 108, or 5 x 108cells / kg. In some embodiments, a dose of CD 19 CAR-expressing cells up to about 1 x 106, 1.1 x 106, 2 x 106, 3.6 x 106, 5 x 106, 1 x 107, 1.8 x 107, 2 x 107, 5 x 107, 1 x 108, 2 x 108, or 5 x 108cells / kg. In some embodiments, a dose of CD 19 CAR-expressing cells comprises about 1.1 X 106- 1.8 X 107cells / kg. In some embodiments, a dose of CD 19 CAR-expressing cells comprises about 1 x 107, 2 x 107, 5 x 107, 1 x 108, 2 x 108, 5 x 108, 1 x 109, 2 x 109, or 5 x 109cells. In some embodiments, a dose of CD 19 CAR-expressing cells comprises at least about 1 x PAT059889-PCT-SEC01

[0156] 107, 2 x 107, 5 x 107, 1 x IO8, 2 x IO8, 5 x IO8, 1 x IO9, 2 x IO9, or 5 x IO9cells. In some embodiments, a dose of CD 19 CAR expressing cells comprises up to about 1 x 107, 2 x l07, 5 x 107, 1 x 108, 2 x 108, 5 x 108, 1 x 109, 2 x 109, or 5 x 109cells. In some embodiments, the CD 19 CAR-expressing cells are rapcabtagene autoleucel.

[0157] The level of CAR-positive cells can be determined according to the methods disclosed in Example 8 of WO / 2021 / 173985. Briefly, for CAR T cells manufactured using a continuous Activated Rapid Manufacturing (ARM) process, e.g., ARM-CD19 CAR T cells, a sentinel vial of cryopreserved cells may be thawed and cultured for up to 5 days and the CAR expression analyzed by flow cytometry. The measurement of CAR expression on, e.g., day 2 or day 3 may be used to determine the dose of viable CAR-positive T cells.

[0158] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient having a myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma- associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, comprising administering to said patient CAR-expressing cells produced as described herein, at a dose of viable CAR-expressing or CAR-positive cells (for example, viable CD 19 CAR-expressing cells, viable CD 19 CAR-positive cells, or any dual CARs thereof) from about 0.5 x 106viable CAR- expressing or CAR-positive cells to about 50 x 106viable CAR-expressing or CAR-positive cells (for example, from about 0.5 x 106viable CD 19 CAR-expressing or CAR-positive cells to about 90 x 106viable CD19 CAR-expressing or CAR-positive cells), e.g. at a dose of viable CAR- expressing or CAR-positive cells (for example, viable CD 19 CAR-expressing cells or viable CD19 CAR-positive cells) from about 2 x 106viable CAR-expressing or CAR-positive cells to about 40 x 106viable CAR-expressing or CAR-positive cells.

[0159] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, comprising administering to said patient CAR-expressing or CAR-positive cells produced as described herein, at a dose of viable CAR-expressing or CAR-positive cells from about 0.5 x 106viable CAR-expressing or CAR-positive cells to about 50 x 106viable CAR-expressing or CAR- PAT059889-PCT-SEC01 positive cells (for example, from about 0.5 x 106viable CD 19 CAR-expressing or CAR-positive cells to about 50 x 106viable CD 19 CAR-expressing or CAR-positive cells).

[0160] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, comprising administering to said patient CAR-expressing or CAR-positive cells produced as described herein at a dose of about 2.5 x 106viable CD19 CAR-expressing, e.g., rapcabtagene autoleucel.

[0161] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, comprising administering to said patient CAR-expressing or CAR-positive cells produced as described herein at a dose of about 7.5 x 106viable CD19 CAR-expressing, e.g., rapcabtagene autoleucel.

[0162] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, comprising administering to said patient CAR-expressing or CAR-positive cells produced as described herein at a dose of about 12.5 x 106viable CD 19 CAR-expressing, e.g., rapcabtagene autoleucel.

[0163] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, comprising administering to said patient CAR-expressing or CAR-positive cells produced as described herein, at a dose of from about 2.5 x 106viable CD 19 CAR-expressing or CAR-positive cells to about 40 x 106viable CD 19 CAR-expressing or CAR-positive cells.

[0164] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, comprising PAT059889-PCT-SEC01 administering to said patient CAR-expressing or CAR-positive cells produced as described herein, at a dose of from about 5 x 106viable CD 19 CAR-expressing or CAR-positive cells to about 12.5 x 106viable CD 19 CAR-expressing or CAR-positive cells.

[0165] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, comprising administering to said patient CAR-expressing or CAR-positive cells produced as described herein, at a dose of from about 25 x 106viable CD 19 CAR-expressing or CAR-positive cells to about 40 x 106viable CD 19 CAR-expressing or CAR-positive cells.

[0166] Evaluating CAR Safety

[0167] In some embodiments of any of the therapeutic methods disclosed herein, the method further involves evaluating the safety of the CAR-expressing cell therapy in a subject. In some embodiments, safety of the CAR-expressing cell therapy is evaluated by measuring or recording one or more of a subject’s vital signs, adverse events experienced by the subject, various laboratory parementers, and / or an electrocardiogram of the subject.

[0168] In some embodiments, the subject does not experience an adverse event of grade 4 or higher. In some embodiments, the subject does not experience cytokine release syndrome (CRS). In some embodiments, the subject does not experience CRS of grade 3 or higher. In some embodiments, the subject does not experience immune effector cell-associated neurotoxicity syndrome (ICANS).

[0169] Methods of Manufacturing

[0170] Provided herein are methods of manufacturing immune effector cells (for example, T cells or NK cells) engineered to express a CAR, for example, a CAR described herein, compositions comprising such cells, and methods of using such cells for treating a disease, such as myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, in a subject. In some embodiments, the methods disclosed herein may manufacture immune effector cells engineered to express a CAR in less than 24 hours. Without wishing to be bound by theory, the methods PAT059889-PCT-SEC01 provided herein preserve the undifferentiated phenotype of T cells, such as naive T cells, during the manufacturing process. These CAR-expressing cells with an undifferentiated phenotype may persist longer and / or expand better in vivo after infusion. In some embodiments, CART cells produced by the manufacturing methods provided herein comprise a higher percentage of stem cell memory T cells, compared to CART cells produced by the traditional manufacturing process, e.g., as measured using scRNA-seq. (FIG. 3). In some embodiments, CART cells produced by the manufacturing methods provided herein comprise a higher percentage of effector T cells, compared to CART cells produced by the traditional manufacturing process, e.g., as measured using scRNA-seq. (FIG. 3). In some embodiments, CART cells produced by the manufacturing methods provided herein better preserve the sternness of T cells, compared to CART cells produced by the traditional manufacturing process. (FIG. 3). In some embodiments, CART cells produced by the manufacturing methods provided herein show a lower level of hypoxia, compared to CART cells produced by the traditional manufacturing process, e.g., as measured using scRNA-seq. (FIG. 3). In some embodiments, CART cells produced by the manufacturing methods provided herein show a lower level of autophagy, compared to CART cells produced by the traditional manufacturing process. (FIG. 3).

[0171] In some embodiments, the CART cells manufactured by the methods disclosed herein may be administered to a subject with minimal ex vivo expansion, for example, less than 1 day, less than 12 hours, less than 8 hours, less than 6 hours, less than 4 hours, less than 3 hours, less than 2 hours, less than 1 hour, or no ex vivo expansion. Accordingly, the methods described herein provide a fast-manufacturing process of making improved CAR-expressing cell products for use in treating a disease in a subject.

[0172] In some embodiments, the present disclosure provides methods of making a population of cells (for example, T cells) that express a chimeric antigen receptor (CAR) comprising: (i) contacting a population of cells (for example, T cells, for example, T cells isolated from a frozen or fresh leukapheresis product from a subject having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies, with (A) an agent that stimulates a CD3 / TCR complex and / or (B) an agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells; (ii) contacting the population of cells (for example, T cells) with a nucleic acid molecule (for PAT059889-PCT-SEC01 example, a DNA or RNA molecule) encoding the CAR, thereby providing a population of cells (for example, T cells) comprising the nucleic acid molecule, and (iii) harvesting the population of cells (for example, T cells) for storage (for example, reformulating the population of cells in cryopreservation media) or administration, wherein: (a) step (ii) is performed together with step

[0173] (i) or no later than 20 hours after the beginning of step (i), for example, no later than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 hours after the beginning of step (i), for example, no later than 18 hours after the beginning of step (i), and step (iii) is performed no later than 26 hours after the beginning of step (i), for example, no later than 22, 23, or 24 hours after the beginning of step (i), for example, no later than 24 hours after the beginning of step (i); (b) step

[0174] (ii) is performed together with step (i) or no later than 20 hours after the beginning of step (i), for example, no later than 12, 13, 14, 15, 16, 17, or 18 hours after the beginning of step (i), for example, no later than 18 hours after the beginning of step (i), and step (iii) is performed no later than 30, 36, or 48 hours after the beginning of step (ii), for example, no later than 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, or 48 hours after the beginning of step (ii); or (c) the population of cells from step (iii) are not expanded, or expanded by no more than 5, 10, 15, 20, 25, 30, 35, or 40%, for example, no more than 10%, for example, as assessed by the number of living cells, compared to the population of cells at the beginning of step (i).

[0175] In some embodiments, the nucleic acid molecule in step (ii) is a DNA molecule. In some embodiments, the nucleic acid molecule in step (ii) is an RNA molecule. In some embodiments, the nucleic acid molecule in step (ii) is on a viral vector, for example, a viral vector chosen from a lentivirus vector, an adenoviral vector, or a retrovirus vector. In some embodiments, the nucleic acid molecule in step (ii) is on a non-viral vector. In some embodiments, the nucleic acid molecule in step (ii) is on a plasmid. In some embodiments, the nucleic acid molecule in step (ii) is not on any vector. In some embodiments, step (ii) comprises transducing the population of cells (for example, T cells) a viral vector comprising a nucleic acid molecule encoding the CAR.

[0176] In some embodiments, the population of cells (for example, T cells) is collected from an apheresis sample (for example, a leukapheresis sample) from a subject having myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies. PAT059889-PCT-SEC01

[0177] In some embodiments, the apheresis sample (for example, a leukapheresis sample) is collected from the subject and shipped as a fresh product or a frozen sample (for example, a cryopreserved sample) to a cell manufacturing facility. Then the T cells (for example, CD4+ T cells and / or CD8+ T cells) are selected from the apheresis sample, for example, using a cell sorting machine (for example, a CliniMACS® Prodigy® device). The selected T cells (for example, CD4+ T cells and / or CD8+ T cells) are then seeded for CART manufacturing using the process described herein. In some embodiments, the selected T cells (for example, CD4+ T cells and / or CD8+ T cells) undergo one or more rounds of freeze-thaw before being seeded for CART manufacturing.

[0178] In some embodiments, cells (for example, T cells) are contacted with anti-CD3 and anti- CD28 antibodies for, for example, 12 hours, followed by transduction with a vector (for example, a lentiviral vector) encoding a CAR. 24 hours after culture initiation, the cells are washed and formulated for storage or administration. Without wishing to be bound by theory, brief CD3 and CD28 stimulation may promote efficient transduction of self-renewing T cells. Compared to traditional CART manufacturing approaches, the activation process provided herein does not involve prolonged ex vivo expansion. (FIG. 3). Similar to the cytokine process, the activation process provided herein also preserves undifferentiated T cells during CART manufacturing.

[0179] In some embodiments, the population of cells is contacted with (A) an agent that stimulates a CD3 / TCR complex and / or (B) an agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells. In some embodiments, the agent that stimulates a CD3 / TCR complex is an agent that stimulates CD3. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor is an agent that stimulates CD28, ICOS, CD27, HVEM, LIGHT, CD40, 4-1BB, 0X40, DR3, GITR, CD30, HM1 , CD2, CD226, or any combination thereof. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor is an agent that stimulates CD28.

[0180] In some embodiments, the agent that stimulates a CD3 / TCR complex is chosen from an antibody (for example, a single-domain antibody (for example, a heavy chain variable domain antibody), a peptibody, a Fab fragment, or a scFv), a small molecule, or a ligand (for example, a naturally existing, recombinant, or chimeric ligand). In some embodiments, the agent that PAT059889-PCT-SEC01 stimulates a costimulatory molecule and / or growth factor receptor is chosen from an antibody (for example, a single-domain antibody (for example, a heavy chain variable domain antibody), a peptibody, a Fab fragment, or a scFv), a small molecule, or a ligand (for example, a naturally existing, recombinant, or chimeric ligand). In some embodiments, the agent that stimulates a CD3 / TCR complex does not comprise a bead. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor does not comprise a bead. In some embodiments, the agent that stimulates a CD3 / TCR complex comprises an anti-CD3 antibody. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor comprises an anti-CD28 antibody. In some embodiments, the agent that stimulates a CD3 / TCR complex comprises an anti-CD3 antibody covalently attached to a colloidal polymeric nanomatrix. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor comprises an anti-CD28 antibody covalently attached to a colloidal polymeric nanomatrix.

[0181] In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor is an agent that stimulates CD28, ICOS, CD27, CD25, 4-1BB, IL6RA, IL6RB, or CD2. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor comprises one or more of a CD28, ICOS, CD27, CD25, 4-1BB, IL6RB, and / or CD2 antigen binding domain, such as but not limited to an anti- CD28, anti-ICOS, anti-CD27, anti-CD25, anti-4-lBB, anti-IL6RA, anti-IL6RB, or anti-CD2 antibody or an antibody fragment comprising one or more CDRs, heavy chain, and / or light chain thereof - such as but not limited to an anti- CD28, anti-ICOS, anti-CD27, anti-CD25, anti-4-lBB, anti-IL6RA, anti-IL6RB, or anti-CD2 antibody provided in Table 27 of WO / 2021 / 173985, hereby incorporated by reference in its entirety. In some embodiments, the agent that stimulates a CD3 / TCR complex and the agent that stimulates a costimulatory molecule and / or growth factor receptor comprise T Cell TransAct™. In some embodiments, the agent that stimulates a CD3 / TCR complex and the agent that stimulates a costimulatory molecule and / or growth factor receptor are comprised in a multispecific binding molecule. In some embodiments, the multispecific binding molecule comprises a CD3 antigen binding domain and a CD28 or CD2 antigen-binding domain.

[0182] In some embodiments, the population of cells is contacted with a nucleic acid molecule encoding a CAR. In some embodiments, the population of cells is transduced with a DNA molecule encoding a CAR. In some embodiments, contacting the population of cells with the PAT059889-PCT-SEC01 nucleic acid molecule encoding the CAR occurs simultaneously with contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0.5 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 20 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 19 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 18 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 17 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 16 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 15 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that PAT059889-PCT-SEC01 stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 14 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 14 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 13 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 12 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 11 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 10 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 9 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 8 hours after the beginning of contacting the PAT059889-PCT-SEC01 population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 7 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 6 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 5 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 4 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 3 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 2 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 1 hour after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid PAT059889-PCT-SEC01 molecule encoding the CAR occurs no later than 30 minutes after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above.

[0183] In some embodiments, the population of cells is harvested for storage or administration. In some embodiments, the population of cells is harvested for storage or administration no later than 72, 60, 48, 36, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, or 18 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 26 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 25 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 24 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 23 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 22 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above.

[0184] In some embodiments, the population of cells is not expanded ex vivo. In some embodiments, the population of cells is expanded by no more than 5, 6, 7, 8, 9, 10, 11, 12, 13, PAT059889-PCT-SEC01

[0185] 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, or 60%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 5%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 10%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 15%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 20%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 25%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 30%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 35%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or PAT059889-PCT-SEC01 growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 40%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above.

[0186] In some embodiments, the activation process is conducted in serum free cell media. In some embodiments, the activation process is conducted in cell media comprising one or more cytokines chosen from: IL-2, IL-15 (for example, hetIL-15 (IL15 / sIL-15Ra)), or IL-6 (for example, IL-6 / sIL-6Ra). In some embodiments, the activation process is conducted in cell media comprising a LSD1 inhibitor. In some embodiments, the activation process is conducted in cell media comprising a MALT1 inhibitor. In some embodiments, the serum free cell media comprises a serum replacement. In some embodiments, the serum replacement is CTS™ Immune Cell Serum Replacement (ICSR). In some embodiments, the level of ICSR can be, for example, up to 5%, for example, about 1%, 2%, 3%, 4%, or 5%.

[0187] In some embodiments, the present disclosure provides methods of making a population of cells (for example, T cells) that express a chimeric antigen receptor (CAR) comprising: (a) providing an apheresis sample (for example, a fresh or cryopreserved leukapheresis sample) collected from a subject with myasthenia gravis (MG), e.g., early-onset MG, late-onset MG, thyoma-associated MG, antibody mediated MG (e.g., AChR+, MuSK+, LRP4+), ocular MG, or MG with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies; (b) selecting T cells from the apheresis sample (for example, using negative selection, positive selection, or selection without beads); (c) seeding isolated T cells at, for example, 1 x 106to 1 x 107cells / mL; (d) contacting T cells with an agent that stimulates T cells, for example, an agent that stimulates a CD3 / TCR complex and / or an agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells (for example, contacting T cells with anti-CD3 and / or anti-CD28 antibody, for example, contacting T cells with TransAct); (e) contacting T cells with a nucleic acid molecule (for example, a DNA or RNA molecule) encoding the CAR (for example, contacting T cells with a virus comprising a nucleic acid molecule encoding the CAR) for, for example, 6-48 hours, for example, 20-28 hours; and (f) washing and harvesting T cells for storage (for example, reformulating T cells in cryopreservation media) or administration. In some embodiments, step (f) is performed no later PAT059889-PCT-SEC01 than 30, 36, or 48 hours after the beginning of step (d) or (e), for example, no later than 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, or 48 hours after the beginning of step (d) or (e).

[0188] In some embodiments of the aforementioned methods, the methods are performed in a closed system. In some embodiments, T cell separation, activation, transduction, incubation, and washing are all performed in a closed system. In some embodiments of the aforementioned methods, the methods are performed in separate devices. In some embodiments, T cell separation, activation and transduction, incubation, and washing are performed in separate devices.

[0189] In some embodiments of the aforementioned methods, the methods further comprise adding an adjuvant or a transduction enhancement reagent in the cell culture medium to enhance transduction efficiency. In some embodiments, the adjuvant or transduction enhancement reagent comprises a cationic polymer. In some embodiments, the adjuvant or transduction enhancement reagent is chosen from: LentiBOOST™ (Sirion Biotech), vectofusin-1, F108 (Poloxamer 338 or Pluronic® F-38), protamine sulfate, hexadimethrine bromide (Polybrene), PEA, Pluronic F68, Pluronic Fl 27, Synperonic or LentiTrans™. In some embodiments, the transduction enhancement reagent is LentiBOOST™ (Sirion Biotech). In some embodiments, the transduction enhancement reagent is Fl 08 (Poloxamer 338 or Pluronic® F-38)

[0190] In some embodiments of the aforementioned methods, the transducing the population of cells (for example, T cells) with a viral vector comprises subjecting the population of cells and viral vector to a centrifugal force under conditions such that transduction efficiency is enhanced. In an embodiment, the cells are transduced by spinoculation.

[0191] In some embodiments of the aforementioned methods, cells (e.g., T cells) are activated and transduced in a cell culture flask comprising a gas-permeable membrane at the base that supports large media volumes without substantially compromising gas exchange. In some embodiments, cell growth is achieved by providing access, e.g., substantially uninterrupted access, to nutrients through convection.

[0192] Pharmaceutical Compositions

[0193] The methods described herein can further include formulating a CD 19 CAR-expressing cell in a pharmaceutical composition. Pharmaceutical compositions may comprise a CD 19 PAT059889-PCT-SEC01

[0194] CAR-expressing cell, for example, a plurality of CD 19 CAR-expressing cells, as described herein, in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents or excipients. Such compositions may comprise buffers such as neutral buffered saline, phosphate buffered saline and the like; carbohydrates such as glucose, mannose, sucrose or dextrans, mannitol; proteins; polypeptides or amino acids such as glycine; antioxidants; chelating agents such as EDTA or glutathione; adjuvants (for example, aluminum hydroxide); and preservatives. Compositions can be formulated, for example, for intravenous administration.

[0195] In some embodiments, the pharmaceutical composition is substantially free of, for example, there are no detectable levels of a contaminant, for example, selected from the group consisting of endotoxin, mycoplasma, replication competent lentivirus (RCL), p24, VSV-G nucleic acid, HIV gag, residual anti-CD3 / anti-CD28 coated beads, mouse antibodies, pooled human serum, bovine serum albumin, bovine serum, culture media components, vector packaging cell or plasmid components, a bacterium and a fungus. In some embodiments, the bacterium is at least one selected from the group consisting of Alcaligenes faecalis, Candida albicans, Escherichia coli, Haemophilus influenza, Neisseria meningitides, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumonia, and Streptococcus pyogenes group A.

[0196] When “an immunologically effective amount” or “therapeutic amount” is indicated, the precise amount of the compositions to be administered can be determined by a physician with consideration of individual differences in age, weight, disease progression, and condition of the patient (subject). It can generally be stated that a pharmaceutical composition comprising the immune effector cells (for example, T cells, NK cells) described herein may be administered at a dosage of about 0.5 x 106to 50 x 106viable CAR-expressing cells, in some instances about 2.5 x 106, 7.5 x 106, or 12.5 x 106viable CAR-expressing cells, including all integer values within those ranges. T cell compositions may also be administered multiple times at these dosages. The cells can be administered by using infusion techniques that are commonly known in immunotherapy (see, for example, Rosenberg et al., New Eng. J. of Med. 319:1676, 1988).

[0197] The administration of the subject compositions may be carried out in any convenient manner. The compositions described herein may be administered to a patient trans arterially, subcutaneously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, by intravenous (i.v.) injection, or intraperitoneally, for example, by intradermal or subcutaneous PAT059889-PCT-SEC01 injection. The compositions of immune effector cells (for example, T cells, NK cells) may be injected directly into a lymph node or site of disease.

[0198] EXAMPLES

[0199] The invention is further described in detail by reference to the following experimental examples. These examples are provided for purposes of illustration only, and are not intended to be limiting unless otherwise specified. Thus, the invention should in no way be construed as being limited to the following examples, but rather, should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.

[0200] Example 1: Description of the Activated Rapid Manufacturing (ARM) process

[0201] In some embodiments, CART cells are manufactured using a continuous Activated Rapid Manufacturing (ARM) process, over approximately 2 days, which will potentially allow for a greater number of less differentiated T cells (T naive and TSCM (stem central memory T) cells) to be returned to a patient for in vivo cellular expansion. The short manufacturing time period allows the early differentiated T cells profile to proliferate in the body for their desired terminal differentiated state rather that in an ex vivo culture vessel.

[0202] In some embodiments, CART cells are manufactured using cryopreserved leukapheresis source material, for example, non-mobilized autologous peripheral blood leukapheresis (LKPK) material. Cryopreserved source material undergoes processing steps for T cell enrichment on the first day of production (Day 0) by means of anti-CD4 / anti-CD8 immunomagnetic system. Positive fraction is then seeded in G-rex culture vessel, activated with an anti-CD3 / CD28 system (TransACT™) and on the same day transduced with a lentiviral vector (LV) encoding a CAR. On the following day, after 20-28 hours of transduction, the T cells are harvested, washed four times, formulated in freezing medium, and then frozen by a Controlled Rate Freezer (CRF). From the start of the process on Day 0 to the initiation of harvest on the following day, cells are cultured for 20 - 28 hours with a target of 24 hours after Day 0 seeding.

[0203] Media for Day 0 were prepared according to Table 1.

[0204] Table 1: Media type and point of use during CART manufacturing PAT059889-PCT-SEC01

[0205] The cryopreserved leukapheresis material is thawed. The thawed cells are diluted with the Rapid Buffer (Table 21) and washed on the CliniMACS® Prodigy® device. The T cells are selected by CliniMACS® CD4 and CD8 microbeads. Once the program is finished for T cell selection (approximately 3h 40 min to 4h 40 min), the reapplication bag containing the cells suspended in Rapid Media (Table 21) are transferred in a transfer pack. A sample is taken for viability and cell count. The cell count and viability data from the positive fraction bag is used to determine the cell concentration when seeding the culture vessel for activation and vector transduction.

[0206] Following positive selection of T cells via the CliniMACS® microbeads (CD4 and CD8), the cells are seeded in the culture vessel, G-Rex. Once the cells are seeded, the activation reagent (TransACT™) is then added to the culture vessel. The cells are then transduced with a lentiviral vector encoding a CAR at a target MOI of 1.0 (0.8-1.2). Following the vector addition, the culture vessel is transported to an incubator where it is incubated for a target of 24 hours (operating range 20-28 hours) at a nominal temperature of 37 °C (operating range 36-38 °C) with nominal 5% CO2 (operating range 4.5-5.5%). Following the incubation, the cells are washed with Harvest Wash Solution (Table 1) four times to remove any non-integrated vector and residual viral particles, as well as any other process related impurities. Then, the cells are eluted and a sample for cell count and viability is taken for testing and the results are used to determine the volume required to re-suspend the cells for final formulation with CryoStor® CS10. The cells are then centrifugated to remove the Harvest Wash Solution and proceed with cryopreservation.

[0207] In some embodiments, the CAR expressed in CART cells binds to CD 19. In some embodiments, the CAR made by the ARM process is rapcabtagene autoleucel. In some PAT059889-PCT-SEC01 embodiments, IL-2 used in the Rapid Media (RM) (Table 1) can be replaced with IL-15, hetlL- 15 (IL-15 / sIL-15Ra), IL-6, or IL-6 / sIL-6Ra.

[0208] Example 2: Phase 1 / 2 study, open-label, multi-center, to assess safety, efficacy and cellular kinetics of rapcabtagene autoleucel cells in participants with treatment resistant myasthenia gravis.

[0209] The purpose of this study is to assess safety, cellular kinetics and preliminary benefit / risk profile of rapcabtagene autoleucel, an autologous CD 19- directed, genetically engineered T cell immunotherapy (CAR-T), as treatment in patients with treatment-resistant generalized myasthenia gravis (gMG).

[0210] Rapcabtagene autoleucel is an autologous CD19-directed CAR-T cell therapy that is comprised of CD4+ / CD8+ T cells that have undergone ex vivo T cell activation and gene modification. Rapcabtagene autoleucel utilizes the FMC63 scFv domain for CD 19 recognition and the same lentiviral vector as tisagenlecleucel (Kymriah, CTL019) and is manufactured via the activated rapid manufacturing (ARM) process. The ARM process reduces the turnaround time compared to traditional manufacturing processes (FIG. 3) and preserves T cell sternness, the ability to self-renew and mature, resulting in a product with greater proliferative potential and fewer exhausted T cells compared to traditionally manufactured CAR-T cells. With ARM, CAR- T cell expansion occurs primarily within a patient’s body (in vivo), eliminating the need for an extended culture time outside of the body (ex vivo). These unique characteristics may lead to better and more durable responses, improved long-term outcomes and a reduced risk of severe adverse events compared to CAR-T cell products manufactured via traditional manufacturing methods. Non-clinical studies show that rapcabtagene autoleucel is a product with potentially superior efficacy, a similar safety profile, and delayed expansion compared to other CD 19- directed CAR-T cell therapy relying on a traditional manufacturing process (e.g. tisagenlecleucel). The expected improved safety and efficacy profile of rapcabtagene autoleucel compared to other CAR-T therapies, offers a rapid and more efficient B cell depletion due to its mode of action and trafficking properties and its ability to also deplete CD 19-positive B cells, including CD20-negative cell subsets such as plasmablasts and a fraction of plasma cells. Rapcabtagene autoleucel also has the advantage of eliminating not only circulating CD 19- expressing cells, but also tissue-resident cells such as B cells in lymph nodes and potentially PAT059889-PCT-SEC01 even thymic B cells. In addition, rapcabtagene autoleucel may induce a long-lasting remission with a decreased treatment burden for the patient and despite short term toxicity risk, better longterm safety.

[0211] Disease

[0212] MG is a rare condition where autoantibodies targeting various components of the postsynaptic neuromuscular junction are produced and can be detected in patient's serum / plasma. These antibodies drive the disease pathogenesis by either blocking neuromuscular signaling / transmission or by complement-mediated damage / lysis of the postsynaptic muscle membrane. The most common target antigens are the acetylcholine receptor (AChR) and the muscle specific tyrosine kinase (MuSK). The disease manifests by varying distributions of muscle weakness that typically worsens after repeated muscle contraction (Gilhus 2016). The clinical manifestation may be heterogeneous, but in its worst manifestation may include the respiratory muscles leading to a condition called myasthenic crisis that requires ventilatory / respiratory support and ICU-level care. Disease severity and distribution may be classified by the Myasthenia Gravis Foundation of America (MGFA) clinical classification (see below), where generalized MG (gMG) corresponds to MFGA class II-V (Jaretzki et al 2000).

[0213] MGFA Clinical Classification

[0214] • Class 0: No evidence of muscle weakness on examination other than weakness of eye closure.

[0215] • Class I: Any ocular muscle weakness; may have weakness of eye closure. All other muscle strength is normal.

[0216] • Class II: Mild weakness affecting muscles other than ocular muscles; may also have ocular muscle weakness of any severity. o Ila. Predominantly affecting limb, axial muscles, or both. May also have lesser involvement of oropharyngeal muscles. o lib. Predominantly affecting oropharyngeal, respiratory muscles, or both. May also have lesser or equal involvement of limb, axial muscles, or both.

[0217] • Class III: Moderate weakness affecting muscles other than ocular muscles; may also have ocular muscle weakness of any severity. o Illa. Predominantly affecting limb, axial muscles, or both. May also have lesser involvement of oropharyngeal muscles. PAT059889-PCT-SEC01 o Illb. Predominantly affecting oropharyngeal, respiratory muscles, or both. May also have lesser or equal involvement of limb, axial muscles, or both.

[0218] • Class IV: Severe weakness affecting muscles other than ocular muscles; may also have ocular muscle weakness of any severity. o IVa. Predominantly affecting limb, axial muscles, or both. May also have lesser involvement of oropharyngeal muscles. o IVb. Predominantly affecting oropharyngeal, respiratory muscles, or both. May also have lesser or equal involvement of limb, axial muscles, or both.

[0219] • Class V: Defined as intubation, with or without mechanical ventilation, except when employed during routine postoperative management. Note: The use of a feeding tube without intubation places the patient in class IVb.

[0220] Overall Study Design

[0221] This is an open-label, single-arm, multi-center, non-confirmatory study to assess safety, efficacy and cellular kinetics of YTB323 in participants with treatment resistant gMG. The cohort design is shown in FIG. 2. Approximately 15 patients will receive treatment with rapcabtagene autoleucel. The study design / participant journey is illustrated in FIG. 1. The total study duration for a participant will be up to 27.5 months. After this, a long-term follow-up will be initiated until 15 years after rapcabtagene autoleucel administration.

[0222] After consenting, participants will first be evaluated for clinical eligibility, and if initially proven to be eligible, the participant will begin tapering of corticosteroids and stopping of immune modulating treatments according to medication washout guidance. Before the planned leukapheresis and CAR-T cell infusion, immunosuppressive treatments will be stopped, and corticosteroid dose will be at the target level - 10 mg daily or equivalent.

[0223] The screening period will occur during days -96 to -42. If deemed clinically eligible up to that point (eligible in terms of inclusion / exclusion criteria, stopping of immunosuppressive medications, and successful corticosteroid taper), leukapheresis will be scheduled (days -42 to - 9). To prevent a clinical worsening during immune treatment washout, patients will be treated with intravenous immune globulin (IVIg) 2 grams / kg divided over 2- 5 days at the time of discontinuation of baseline immunosuppressive drugs. Whether IVIg will be administered over a 2-5 day course and the precise timing of administration of IVIg during tapering of corticosteroids PAT059889-PCT-SEC01 will be at the discretion of the investigator based clinical status and BL corticosteroid dose. If a patient experiences clinical worsening subsequent to this and prior to receiving rapcabtagene autoleucel, they will be treated according to standard of care practice, but preferably with additional courses of IVIg. If tapering requires longer time than the specified duration of the screening period, then the screening window may be extended. Once the leukapheresis product has been confirmed to be suitable for rapcabtagene autoleucel manufacturing, the manufacturing process will commence.

[0224] After the final product has been confirmed to be available, participants will receive lymphodepleting therapy (up to 9 days prior to Day 1). Following pre-infusion check on Day 1 and premedication, rapcabtagene autoleucel will be given as a single infusion on Day 1. Each participant will be hospitalized until at least Day 14 following rapcabtagene autoleucel infusion. Participants will be closely monitored for any safety events for the first 2 months (twice a week visits for the first 5 weeks followed by weekly visits up to end of Month 2 (Day 56). After that, the visit frequency will be reduced to monthly visits up to Month 6 and quarterly visits thereafter up to Month 24 (FIG. 1). End of study visit for a participant in this study / protocol will be completed at Month 24 but participants will continue to be followed in the long-term follow-up (LTFU).

[0225] Study Population

[0226] The study population is individuals with anti-AChR or Anti-MuSK-seropositive gMG that are treatment-resistant to established immune therapies. Treatment-resistant gMG as defined by: MG-ADL score > 6 at screening despite adequate treatment trials with at least two different non-steroidal immunosuppressive drugs given at adequate doses and duration of therapy. Specifically, they must have failed treatment with at least one oral immunomodulatory or steroid-sparing drug (e.g., azathioprine, mycophenolate mofetil, tacrolimus) and either one approved anti-C5 complement antibody (e.g., eculizumab, ravulizumab) or an approved FcRn antagonist or rituximab or required two or more rescue therapies (plasma exchange / immunoadsorption or IVIg) for myasthenic worsening / crisis in the 12 months before screening.

[0227] Study Treatment

[0228] Eligible participants undergo the following sequence of events prior to rapcabtagene autoleucel administration: (1) course of intravenous immunoglobulin (IVIg) to prevent disease PAT059889-PCT-SEC01 exacerbation due to immunotherapy washout / corticosteroid taper, (2) leukapheresis, (3) pre- lymphodepletion evaluation, (4) lymphodepletion, (5) premedication, and (6) pre- rapcabtagene autoleucel administration check.

[0229] Prior to rapcabtagene autoleucel administration, each participant should undergo lymphodepleting therapy with at least one day of rest immediately after lymphodepletion and prior to rapcabtagene autoleucel infusion. Lymphodepleting therapy starts within one week before rapcabtagene autoleucel injection, which means that rapcabtagene autoleucel is injected 2 to 6 days after lymphodepleting therapy is completed. Lymphodepleting therapy may be repeated in the case rapcabtagene autoleucel has been delayed by more than 2 weeks. The lymphodepleting therapy regime is as follows: (1) fludarabine administered 25 mg / m2intravenously [i.v.] daily for 3 doses (for participants with renal impairment, the dose may be reduced as per local approved labels of fludarabine), and (2) cyclophosphamide administered 250 mg / m2i.v. daily for 3 doses starting with the first dose of fludarabine.

[0230] All participants are pre-medicated with acetaminophen (paracetamol, 500-1000 mg, orally) and diphenhydramine (25-50 mg, i.v. or orally) or another Hl antihistamine approximately 30 to 60 minutes prior to injection. These medications can be repeated every 6 hours as needed. Non-steroidal anti-inflammatory medication may be prescribed if the participant continues to have fever not relieved with acetaminophen (paracetamol). Steroids should not be used for premedication.

[0231] Rapcabtagene autoleucel treatment will consist of a single intravenous (i.v.) administration of a dose of 12.5 x 106CAR-positive viable T cells with the acceptable release specifications.

[0232] Safety, Pharmacokinetic (PK) and Efficacy Assessments

[0233] The primary aim of the study is to assess the safety of a single dose of rapcabtagene autoleucel in gMG patients. For the assessment of safety, all safety data including laboratory measurements, vital signs, adverse events (AEs), AEs of special interest (AESIs), serious adverse events (SAEs), ECG measurements and neurological status are considered as primary safety endpoints. PAT059889-PCT-SEC01

[0234] The primary safety question of interest is to assess the frequency and severity of AEs of special interest (AESI) in the 8 weeks following rapcabtagene autoleucel (anti-CD19 CAR T- cell) infusion in patients with treatment-resistant gMG. The AESI are defined as:

[0235] • CRS: Cytokine Release Syndrome

[0236] • ICANS: Immune effector Cell-Associated Neurotoxic Syndrome

[0237] The primary estimand is described by the following attributes:

[0238] • Population: patients with treatment-resistant generalized myasthenia gravis (gMG) with antibodies against AChR or MuSK as defined by an MG-ADL of >6 (>50% nonocular) despite treatment with an immunomodulating / immunosuppressive drug as soc.

[0239] • Primary Variable / Endpoint: Incidence and severity of selected AESI (as defined above) within 56 days after rapcabtagene autoleucel CAR T-cell infusion.

[0240] • Treatment of interest: pre-treatment / treatment procedures ending with rapcabtagene autoleucel administration.

[0241] • Inter current events (ICEs): Lost to follow-up (FU), significant disease exacerbation (worsening of >3 points in QMG), patient withdrawal, and initiation of new therapy for worsening of disease will be addressed with treatment policy strategy. Summary measure: frequency and severity of AESIs in pre-specified time interval of 56 days following rapcabtagene autoleucel administration.

[0242] Several clinical and patient reported outcome measures are included in this study for all participants. The impact of gMG on various aspects of a gMG patient's health status will be assessed by the following measures:

[0243] Clinical outcome assessments (COAs)

[0244] • Myasthenia Gravis-Activities of Daily Living (MG-ADL) (Wolfe et al 1999)

[0245] • Quantitative Myasthenia Gravis Scale (QMG) (Barohn et al 1998)

[0246] • MGFA Post-Intervention Status (PIS) (Jaretzki et al 2000)

[0247] Patient reported outcomes (PROs)

[0248] • Myasthenia gravis-Quality of Life 15-revised (MG-QoL15r) (Burns et al 2011)

[0249] • Short Form Health Survey (SF-36) (Boldingh et al 2015) Objectives and related endpoints are summarized in Table 2.

[0250] Table 2 Objectives and related endpoints PAT059889-PCT-SEC01

[0251] Objective(s) Endpoint(s)

[0252] Primary objective(s) Endpoint(s) for primary objective(s)

[0253] • To assess the safety of • Occurrence, severity, and frequency of Adverse Events rapcabtagene autoleucel in patients (AEs) (including CRS and ICANs) and change from with treatment-resistant generalized baseline in safety parameters including, but not limited myasthenia gravis (gMG) with to: Vital signs, laboratory parameters, ECG, and antibodies against AChR or MuSK neurological status

[0254] Secondary objective(s) Endpoint(s) for secondary objective(s)

[0255] • To characterize the in vivo cellular • rapcabtagene autoleucel transgene levels by qPCR kinetics (pharmacokinetics, PK) of over time in peripheral blood; cellular kinetics rapcabtagene autoleucel in parameters (Cmax, AUC, Tmax, Clast, Tlast) peripheral blood by quantitative polymerase chain reaction (qPCR)

[0256] • To characterize the incidence and • Pre-existing and treatment induced immunogenicity prevalence of pre-existing and (cellular, humoral, neutralizing antibodies) of treatment induced immunogenicity rapcabtagene autoleucel (cellular and humoral) of rapcabtagene autoleucel

[0257] • To assess the effect of • At various timepoints: rapcabtagene autoleucel on gMG • Change from BL of MG-ADL score

[0258] • Change from BL of QMG total score disease severity

[0259] • Whether or not patient achieves a >3-point reduction of

[0260] QMG total score sustained for 6 months post BL

[0261] • Whether or not patient achieves a >2-point reduction of

[0262] MG-ADL score sustained for 6 months post BL

[0263] • Whether or not a patient achieves MGFA Post-intervention Status (PIS) of minimal manifestations (MM) or better and sustained for 6 months post BL

[0264] • To evaluate the feasibility of the 1 . Manufacture success (defined as manufacturing process for patients meeting release specifications and with gMG target dose)

[0265] Several clinician and patient reported outcomes are included in this study for all participants:

[0266] • Myasthenia Gravis-Activities of Daily Living (MG-ADL) PAT059889-PCT-SEC01

[0267] The Myasthenia Gravis Activities of Living (MG-ADL) scale is an 8-item patient- reported scale that measures myasthenia gravis (MG) symptoms and functional status, to be administered by physician or trained study evaluator.

[0268] The MG-ADL is an outcome measure assessing MG symptoms and functional activities related to activities of daily living (Wolfe et al 1999). Each of the items is scored from 0 (normal) to 3 (most severe), providing a total MG-ADL score ranging from 0 to 24, where higher scores indicate greater severity of symptoms. The MG-ADL is composed of items related to patients’ assessment of functional disability secondary to ocular (two items), bulbar (three items), respiratory (one item), and gross motor or limb impairment (two items). Items are linearly scored and not weighted, with each item ranging from 0 to 3 for a total score range of 0 to 24. The MG-ADL is easy to administer, is quick to complete (<10 min), and can be used in routine clinical practice or in clinical trials. Change in this scale has been used in pivotal trials as primary outcome parameter to assess MG disease severity.

[0269] A two-point reduction in MG-ADL total score optimally indicates improvement for patients with MG based on a receiver operator characteristic curve approach. This threshold of a two point difference is recognized as a definition of responder with clinically important improvement.

[0270] • Quantitative Myasthenia Gravis Scale (QMG)

[0271] The QMG score is a standardized quantitative strength scoring system developed specifically for MG. The QMG has been validated and has been used in several previous MG trials.

[0272] The QMG is a 13 -item direct physician assessment scoring system that quantifies disease severity based on impairments of body functions and structures (Barohn et al 1998). Each item is quantitatively assessed and scored from 0 to 3 (where 3 represents the most severe), providing a total QMG score ranging from 0 to 39. The QMG is composed of the following items: ocular (two items), facial (one item), bulbar (two items), gross motor (six items), axial (one item), and respiratory (one item). The original study by Barohn et al (1998) that designed the version of the QMG in use today, identified a minimally important difference (MID) of 2.6 points. Some have suggested that the MID should be higher in patients with higher baseline QMG scores. An MID of 3.5 has been used in previous MG trails (Barnett et al 2018).

[0273] • The Myasthenia Gravis Foundation of America Post-Intervention Status (MGFA-PIS) PAT059889-PCT-SEC01

[0274] The MGFA Post-Intervention Status is a physician-assessed determination of the overall clinical state of an MG patient at any time after initiation of treatment for MG. The MGFA-PIS is designed to assess the clinical state of MG patients after they have received treatment for MG (Jaretzki et al 2000). It provides the physician’s global assessment of the patient’s clinical status.

[0275] • Myasthenia Gravis-Quality of Life 15 revised (MG-QoL- 15r

[0276] MG-QoL15r is a 15-question scale designed to assess aspects of life related to myasthenia gravis. There is an appropriate correlation between the MG-QOL 15r and other MG- specific scales (MGC, MG-ADL, and QMG score).

[0277] The MG-QoL15r allows clinicians to estimate a patient’s quality of life relevant to MG (Burns et al 2011). Items on the MG-QoL15r relate to physical, social, and psychological components and are scored from 0 (not at all) to 4 (quite a bit). The cumulative scores range from 0 to 60, with higher scores representing worse quality of life. The MG-QoL15r was constructed based on the most relevant and responsive items from the 60-item version of the questionnaire, with the goal of having a quick, easy-to-use, and easy-to-interpret questionnaire. The MG-QOL 15r has construct validity in the clinical practice setting and represents an efficient and valuable tool for assessing quality of life for patient with MG. However, the magnitude of change required to indicate improvement or worsening is variable and depends on MG severity. The MG-QOL 15r can be completed by the patient or administered by the physician or trained clinic personnel or study coordinator.

[0278] • Short Form Health Survey (SF-36)

[0279] The Short Form Health Survey (SF-36 v2) is a widely used and extensively studied instrument to measure health-related quality of life among healthy participants and participants with acute and chronic conditions. It consists of eight subscales that can be scored individually: Physical Functioning, Role-Physical, Bodily Pain, General Health, Vitality, Social Functioning, Role-Emotional, and Mental Health. Two overall summary scores, the Physical Component Summary (PCS) and the Mental Component Summary (MCS) also can be computed. The SF-36 has proven useful in monitoring general and specific populations, comparing the relative burden of different diseases, differentiating the health benefits produced by different treatments, and in screening individual participants. PAT059889-PCT-SEC01

[0280] The clinical assessments chosen are very standard outcome measures in gMG and have been used in clinical trials of experimental therapeutics in the disease. Several have been validated and used in clinical trials to obtain regulatory approval (MG-ADL, QMG score). In recent years, regulatory authorities have emphasized the use of patient-reported outcomes as a primary efficacy parameter in clinical trials. Accordingly, several successful registrational trials in MG have utilized patient-reported outcomes (MG-ADL) as a primary endpoint (Howard et al 2023, Howard et al 2021, Howard et al 2017). Symptoms fluctuate in gMG; hence, objective physical assessments may not necessarily reflect patients' experienced symptom burden. Consequently, patient-reported outcomes are preferred as endpoints in MG trials and are employed in this trial (eg.MG-QOLl 5r).

[0281] Pharmacokinetics and Immunogenicity

[0282] For pharmacokinetic (PK) analysis, serial blood samples will be collected at specific time points to measure rapcabtagene autoleucel cellular kinetics in peripheral blood by flow cytometry and by qPCR, and to measure humoral and cellular immunogenicity, including neutralizing antibodies. Analytical methods for PK and immunogenicity assessments are listed in Table 3.

[0283] Table 3. Analytical methods associated with the PK and immunogenicity assessments PAT059889-PCT-SEC01

[0284] The absolute number of CD 19+ B cells in peripheral blood will be used as the PD marker to characterize B cell depletion resulting from rapcabtagene autoleucel treatment. The CD 19+ B cell counts will be measured by flow cytometry. The flow cytometry analysis will be performed using a validated panel that also includes the analysis of T cells and NK cells (TBNK panel).

[0285] EQUIVALENTS

[0286] The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety. While this invention has been disclosed with reference to certain embodiments, it is apparent that further embodiments and variations of this invention may be devised by others skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

Claims

PAT059889-PCT-SEC01What is claimed is:

1. A method of treating a subject diagnosed with myasthenia gravis comprising administering rapcabtagene autoleucel to the subject.

2. The method of claim 1, wherein the myasthenia gravis is early-onset myasthenia gravis, late-onset myasthenia gravis , thyoma-associated myasthenia gravis , antibody mediated myasthenia gravis (e.g., AChR+, MuSK+, LRP4+), ocular myasthenia gravis , or myasthenia gravis with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies.

3. The method of claim 1 or 2, wherein the myasthenia gravis is treatment resistant.

4. The method of claims 1-3, wherein the Myasthenia Gravis Activities of Living (MG-ADL) score of the subject is reduced by at least 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points, 18 points, 19 points, 20 points, 21 points, 22 points, 23 points, or 24 points from baseline.

5. The method of claim 4, wherein the reduction in the MG-ADL score is sustained for at least 3 months, 6 months, 9 months, 12 months, 18 months, or 24 months.

6. The method of claim 5, wherein the MG-ADL score is reduced by at least 2 points from baseline and the reduction is sustained for at least 6 months.

7. The method of any of claims 1-6, wherein the Quantitative Myasthenia Gravis Scale (QMG) score of the subject is reduced by at least 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points, 18 points, 19 points, 20 points, 21 points, 22 points, 23 points, 24 points, 25 points, 26 points, 27 points, 28 points, 29 points, 30 points, 31 points, 32 points, 33 points, 34 points, 35 points, 36 points, 37 points, 38 points, or 39 points from baseline.PAT059889-PCT-SEC018. The method of claim 7, wherein the reduction GMQ score is sustained for at least3 months, 6 months, 9 months, 12 months, 18 months, or 24 months.

9. The method of claim 8, wherein the GMQ score is reduced by at least 3 points from baseline and sustained for at least 6 months.

10. The method of any of claims 1-9, wherein the Myasthenia Gravis Foundation of America Post-Intervention Status (MGFA-PIS) of the subject is improved from baseline.

11. The method of claim 10, wherein the subject achieves a MGFA-PIS of minimal manifestations, pharmacologic remission, or complete stable remission.

12. The method of claim 11, wherein the minimal manifestation is MM-3, MM-2, MM-1, or MM-0.

13. The method of claim 12, wherein improved MGFA-PIS is sustained for at least 3 months, 6 months, 12 months, 18 months, or 24 months.

14. The method of claim 13, wherein the subject achieves a MGFA-PIS of minimal manifestations, and that status is sustained for at least 6 months.

15. The method of claim 13, wherein the subject achieves a MGFA-PIS of pharmacologic remission, and that status is sustained for at least 6 months.

16. The method of claim 13, wherein the subject achieves a MGFA-PIS of complete stable remission, and that status is sustained for at least 6 months.

17. The method of claim 1-16, wherein the subject has an increase in the 36-Item Short Form Survey (SF-36) score of at least 5 points, 10 points, 15 points, 20 points, 25 points, 30 points, 35 points, 40 points, 45 points, or 50 points from baseline.PAT059889-PCT-SEC0118. The method of any of claims 1-17, wherein the increase in SF-36 score is sustained for at least 3 months, 6 months, 12 months, 18 months, or 24 months.

19. The method of claim 11, wherein the response is sustained for at least 3 months, 6 months, 12 months, or 18 months.

20. The method of any of claims 1-19, wherein the Myasthenia Gravis-Quality of Life 15 revised (MG-QoL-15r) score of the subject is decreased by at least 5 points, 10 points, 15 points, 20 points, 25 points, 30 points, 35 points, 40 points, 45 points, 50 points, 55 points, or 60 points from baseline.

21. The method of claim 20, wherein the decrease in MG-QoL-15r score is sustained for at least 3 months, 6 months, 9 months, 12 months, 18 months, or 24 months.

22. The method of any of claims 1-21, wherein the autoantibody levels of the subject are decreased from baseline.

23. The method of claim 22, wherein the autoantibody is one or more of AChR, LRP4, or MuSK antibodies.

24. The method of claim 23, wherein the decrease in autoantibody level is sustained for at least 3 months, 6 months, 9 months, 12 months, 18 months, or 24 months.

25. The method of any of claims 1-24, wherein the rapcabtagene autoleucel is administered at a dose of about 0.5 x 106- 1.25 x 109viable CAR-expressing cells.

26. The method of claim 25, wherein the rapcabtagene autoleucel is administered at a dose of 2.5 x 106, 7.5 x 106, or 12.5 x 106viable CAR-expressing cells.PAT059889-PCT-SEC0127. The method of claim 25, wherein the rapcabtagene autoleucel is administered at a dose of 12.5 x 106viable CAR-expressing cells.

28. The method of claim 25, wherein the rapcabtagene autoleucel is administered at a dose of 2.5 x 106viable CAR-expressing cells29. The method of claim 25, wherein the rapcabtagene autoleucel is administered at a dose of 7.5 x 106viable CAR-expressing cells.

30. The method of any of claims 1-29, wherein the rapcabtagene autoleucel is administered in combination with a second therapy.

31. A method of treating a subject diagnosed with myasthenia gravis comprising administering to the subject a population of cells engineered to express a CD 19 CAR, said population comprising:(i) about the same percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;(ii) a change within about 5% to about 10% of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, for example, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;(iii) an increased percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, for example, increased by at least 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, or 3-fold, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;(iv) about the same percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, as compared to the percentage ofPAT059889-PCT-SEC01 central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;(v) a change within about 5% to about 10% of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;(vi) a decreased percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, for example, decreased by at least 20, 25, 30, 35, 40, 45, or 50%, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;(vii) about the same percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR;(viii) a change within about 5% to about 10% of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR; or(ix) an increased percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR.

32. The method of claim 31, wherein the myasthenia gravis is early-onset myasthenia gravis, late-onset myasthenia gravis , thyoma-associated myasthenia gravis , antibody mediated myasthenia gravis (e.g., AChR+, MuSK+, LRP4+), ocular myasthenia gravis , or myasthenia gravis with no detectable known autoantibodies, e.g., AChR, LRP4, and / or MuSK antibodies.PAT059889-PCT-SEC0133. The method of claim 31 or 32, wherein the myasthenia gravis is treatment resistant.

34. The method of claims 31-33, wherein the Myasthenia Gravis Activities of Living (MG-ADL) score of the subject is reduced by at least 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points, 18 points, 19 points, 20 points, 21 points, 22 points, 23 points, or 24 points from baseline.

35. The method of claim 34, wherein the reduction in the MG-ADL score is sustained for at least 3 months, 6 months, 9 months, 12 months, 18 months, or 24 months.

36. The method of claim 35, wherein the MG-ADL score is reduced by at least 2 points from baseline and the reduction is sustained for at least 6 months.

37. The method of any of claims 31-36, wherein the Quantitative Myasthenia Gravis Scale (QMG) score of the subject is reduced by at least 2 points, 3 points, 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 11 points, 12 points, 13 points, 14 points, 15 points, 16 points, 17 points, 18 points, 19 points, 20 points, 21 points, 22 points, 23 points, 24 points, 25 points, 26 points, 27 points, 28 points, 29 points, 30 points, 31 points, 32 points, 33 points, 34 points, 35 points, 36 points, 37 points, 38 points, or 39 points from baseline.

38. The method of claim 37, wherein the reduction GMQ score is sustained for at least 3 months, 6 months, 9 months, 12 months, 18 months, or 24 months.

39. The method of claim 38, wherein the GMQ score is reduced by at least 3 points from baseline and sustained for at least 6 months.

40. The method of any of claims 31-39, wherein the Myasthenia Gravis Foundation of America Post-Intervention Status (MGFA-PIS) of the subject is improved from baseline.PAT059889-PCT-SEC0141. The method of claim 40, wherein the subject achieves a MGFA-PIS of minimal manifestations, pharmacologic remission, or complete stable remission.

42. The method of claim 41, wherein the minimal manifestation is MM-3, MM-2, MM-1, or MM-0.

43. The method of claim 42, wherein improved MGFA-PIS is sustained for at least 3 months, 6 months, 12 months, 18 months, or 24 months.

44. The method of claim 43, wherein the subject achieves a MGFA-PIS of minimal manifestations, and that status is sustained for at least 6 months.

45. The method of claim 43, wherein the subject achieves a MGFA-PIS of pharmacologic remission, and that status is sustained for at least 6 months.

46. The method of claim 43, wherein the subject achieves a MGFA-PIS of complete stable remission, and that status is sustained for at least 6 months.

47. The method of claim 31-46, wherein the subject has an increase in the 36-Item Short Form Survey (SF-36) score of at least 5 points, 10 points, 15 points, 20 points, 25 points, 30 points, 35 points, 40 points, 45 points, or 50 points from baseline.

48. The method of any of claims 31-47, wherein the increase in SF-36 score is sustained for at least 3 months, 6 months, 12 months, 18 months, or 24 months.

49. The method of claim 48, wherein the response is sustained for at least 3 months, 6 months, 12 months, or 18 months.

50. The method of any of claims 31-49, wherein the Myasthenia Gravis-Quality of Life 15 revised (MG-QoL-15r) score of the subject is decreased by at least 5 points, 10 points, 15PAT059889-PCT-SEC01 points, 20 points, 25 points, 30 points, 35 points, 40 points, 45 points, 50 points, 55 points, or 60 points from baseline.

51. The method of claim 50, wherein the decrease in MG-QoL-15r score is sustained for at least 3 months, 6 months, 9 months, 12 months, 18 months, or 24 months.

52. The method of any of claims 31-51, wherein the autoantibody levels of the subject are decreased from baseline.

53. The method of claim 52, wherein the autoantibody is one or more of AChR, LRP4, or MuSK antibodies.

54. The method of claim 53, wherein the decrease in autoantibody level is sustained for at least 3 months, 6 months, 9 months, 12 months, 18 months, or 24 months.

55. A method of reducing the MG-ADL of a subject with myasthenia gravis, the method comprising administering rapcabtagene autoleucel to the subject at a dose of 2.5-, 7.5-, or 12.5 x 10A6 viable CAR-expressing cells.

56. A method of increasing the SF-36 score of a subject with myasthenia gravis, the method comprising administering rapcabtagene autoleucel to the subject at a dose of 2.5-, 7.5-, or 12.5 x 10A6 viable CAR-expressing cells.

57. A method of reducing the MG-QoL-15r of a subject with myasthenia gravis, the method comprising administering rapcabtagene autoleucel to the subject at a dose of 2.5-, 7.5-, or 12.5 x 10A6 viable CAR-expressing cells.

58. A method of reducing the QMG of a subject with relapsing multiple sclerosis, the method comprising administering rapcabtagene autoleucel to the subject at a dose of 2.5-, 7.5-, or 12.5 x 10A6 viable CAR-expressing cells.PAT059889-PCT-SEC0159. A method of reducing the autoantibody level of a subject with relapsing multiple sclerosis, the method comprising administering rapcabtagene autoleucel to the subject at a dose of 2.5-, 7.5-, or 12.5 x 10A6 viable CAR-expressing cells.

60. Rapcabtagene autoleucel for use in a method of treating a subject having myasthenia gravis, said method comprising administering to the subject rapcabtagene autoleucel at a dose of 2.5-, 7.5-, or 12.5 x 10A6 cells.

61. A pharmaceutical composition comprising the rapcabtagene autoleucel of claim 60 and a pharmaceutically acceptable carrier.

62. A method of treating a subject with myasthenia gravis comprising administering to the subject rapcabtagene autoleucel, wherein the rapcabtagene autoleucel is made by a method comprising:(i) contacting a population of T-cells derived from the subject with myasthenia gravis with an agent that stimulates a CD3 / TCR complex;(ii) contacting the population of T-cells with a nucleic acid molecule encoding a CD 19 CAR, thereby providing a population T cells comprising the CD 19 CAR(iii) harvesting the population of T-cells for storage or administration, wherein(a) step (ii) is performed together with step (i) or no later than 20 hours after the beginning of step (i) and step (iii) is performed no later than 30 hours after the beginning of step (i);(b) step (ii) is performed together with step (i) or no later than 20 hours after the beginning of step (i) and step (iii) is performed no later than 30 hours after the beginning of step (ii); or,(c) the population of cells from step (iii) are not expanded as assessed by the number of living cells compared to the population of cells at the beginning of step (i).

63. The method of claim 62, wherein:PAT059889-PCT-SEC01(a) the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) is the same as or differs by no more than 5 or 10% from the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the population of cells at the beginning of step (i);(b) the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) is increased by, for example, at least 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, or 3-fold, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the population of cells at the beginning of step (i);(c) the percentage of CAR-expressing naive T cells, for example, CAR-expressing CD45RA+ CD45RO- CCR7+ T cells in the population of cells increases during the duration of step (ii), for example, increases by, for example, at least 30, 35, 40, 45, 50, 55, or 60%, between 18-24 hours after the beginning of step (ii); or(d) the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) does not decrease, or decreases by no more than 5 or 10%, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the population of cells at the beginning of step (i).

64. The method of any one of claims 62 or 63, wherein:(a) the population of cells from step (iii) shows a higher percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells (for example, at least 10, 20, 30, or 40% higher), compared with cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(b) the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) is higher (for example, at least 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, or 3-fold higher) than the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after thePAT059889-PCT-SEC01 beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(c) the percentage of CAR-expressing naive T cells, for example, CAR-expressing CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) is higher (for example, at least 4, 6, 8, 10, or 12-fold higher) than the percentage of CAR-expressing naive T cells, for example, CAR-expressing CD45RA+ CD45RO- CCR7+ T cells, in cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(d) the population of cells from step (iii) shows a higher percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells (for example, at least 10, 20, 30, or 40% higher), compared with cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days;(e) the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) is higher (for example, at least 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, or 3-fold higher) than the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days; or(f) the percentage of CAR-expressing naive T cells, for example, CAR-expressing CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) is higher (for example, at least 4, 6, 8, 10, or 12-fold higher) than the percentage of CAR-expressing naive T cells, for example, CAR-expressing CD45RA+ CD45RO- CCR7+ T cells, in cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days.

65. The method of any one of claims 62-64, wherein:PAT059889-PCT-SEC01(a) the percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells, in the population of cells from step (iii) is the same as or differs by no more than 5 or 10% from the percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells, in the population of cells at the beginning of step (i);(b) the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells, in the population of cells from step (iii) is reduced by at least 20, 25, 30, 35, 40, 45, or 50%, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells, in the population of cells at the beginning of step (i);(c) the percentage of CAR-expressing central memory T cells, for example, CAR- expressing CCR7+CD45RO+ cells, decreases during the duration of step (ii), for example, decreases by, for example, at least 8, 10, 12, 14, 16, 18, or 20%, between 18-24 hours after the beginning of step (ii); or(d) the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells, in the population of cells from step (iii) does not increase, or increases by no more than 5 or 10%, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells, in the population of cells at the beginning of step (i).

66. The method of any one of claims 62-65, wherein:(a) the population of cells from step (iii) shows a lower percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells (for example, at least 10, 20, 30, or 40% lower), compared with cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(b) the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells in the population of cells from step (iii) is lower (for example, at least 20, 30, 40, or 50% lower) than the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells, in cells made by anPAT059889-PCT-SEC01 otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(c) the percentage of CAR-expressing central memory T cells, for example, CAR- expressing CCR7+CD45RO+ T cells in the population of cells from step (iii) is lower (for example, at least 10, 20, 30, or 40% lower) than the percentage of CAR-expressing central memory T cells, for example, CAR-expressing CCR7+CD45RO+ T cells, in cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(d) the population of cells from step (iii) shows a lower percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells (for example, at least 10, 20, 30, or 40% lower), compared with cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days;(e) the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells in the population of cells from step (iii) is lower (for example, at least 20, 30, 40, or 50% lower) than the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells, in cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days; or(f) the percentage of CAR-expressing central memory T cells, for example, CAR- expressing CCR7+CD45RO+ T cells in the population of cells from step (iii) is lower (for example, at least 10, 20, 30, or 40% lower) than the percentage of CAR-expressing central memory T cells, for example, CAR-expressing CCR7+CD45RO+ T cells, in cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days.

67. The method of any one of claims 62-66, wherein:PAT059889-PCT-SEC01(a) the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor 0+CCR7+CD62L+ T cells, in the population of cells from step (iii) is increased, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells at the beginning of step (i);(b) the percentage of CAR-expressing stem memory T cells, for example, CAR- expressing CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells from step (iii) is increased, as compared to the percentage of CAR-expressing stem memory T cells, for example, CAR-expressing CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells at the beginning of step (i);(c) the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells from step (iii) is higher than the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i); or(d) the percentage of CAR-expressing stem memory T cells, for example, CAR- expressing CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells from step (iii) is higher than the percentage of CAR-expressing stem memory T cells, for example, CAR-expressing CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(e) the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells from step (iii) is higher than the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days; or(f) the percentage of CAR-expressing stem memory T cells, for example, CAR- expressing CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells from step (iii) is higher than the percentage of CAR-expressing stem memory T cells, forPAT059889-PCT-SEC01 example, CAR-expressing CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days.

68. A method of depleting the B cells of a subject with myasthenia gravis, the method comprising administering rapcabtagene autoleucel to the subject at a dose of 2.5-, 7.5-, or 12.5 x 10A6 viable CAR-expressing cells.

69. The method of any of claims 1-68, wherein the subject has experienced breakthrough disease progression while on a therapy.

70. The method of claim 69, wherein the therapy is a non-steroidal immunosuppressive therapy, an anti-C5 complement therapy, FcRN antagonist therapy, or a biologic.

71. The method of claim 70, wherein the therapy is azathioprine, mycophenolate mofetil, tarolimus, eculizumab, ravulizumab, or rituximab.

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