Allogenic car-t for the treatment of cancer

Abstract

Provided herein are, inter alia, are methods for producing expanded population of mycophenolate resistant CAR-T cells. The methods provided herein are, inter alia, useful for producing an expanded population of mycophenolate resistant CAR-T cells for the treatment of cancer. Also provided herein are, inter alia, compositions including the expanded population of mycophenolate resistant CAR-T cells produced by the methods described herein.

Description

PATENT Attorney Docket No.048537-668001WO ALLOGENIC CAR-T FOR THE TREATMENT OF CANCER RELATED APPLICATIONS

[0001] This patent application claims the benefit of U.S. Provisional Patent Application No. 63 / 709,347, filed on October 18, 2024, the contents of which are incorporated herein by reference for all purposes, including all text, tables and drawings. SEQUENCE LISTING

[0002] The material in the accompanying Sequence Listing is hereby incorporated by reference in its entirety and for all purposes. The accompanying file, named “048537- 668001WO_SL_ST26.xml” was created on October 17, 2025, and is 5,078 bytes in size. BACKGROUND

[0003] Currently, the approved CAR-T products are generated from a patient’s own cells. This requires isolation of target T-cells, transduction with a CAR expression vector, and expansion. This process is difficult to complete, labor intensive, time consuming, and does not always generate a suitable product to treat the patient. Allogeneic CAR-Ts offer an off-the-shelf therapeutic that eliminates most of the issues relative to the employment of autologous CAR-T cell products. Provided herein, inter alia, are compositions and methods of use thereof to address these and other problems in the art. BRIEF SUMMARY

[0004] In an aspect is provided a method of producing an expanded population of mycophenolate resistant CAR-T cells, the method including: (i) contacting a plurality of T cells with a first expression vector including a first nucleic acid sequence encoding a chimeric antigen receptor (CAR) thereby producing a plurality of CAR-T cells and expanding said plurality of CAR-T cells for a first period of time thereby producing an expanded population of CAR-T cells, wherein the first period of time is at least 14 days; (ii) contacting said expanded population of CAR-T cells with a second expression vector including a second nucleic acid encoding a mutated IMPDH2 protein thereby producing a plurality of IMPDH2 mutant CAR-T cells, wherein said mutated IMPDH2PATENT Attorney Docket No.048537-668001WO protein confers mycophenolate resistance to said plurality of IMPDH2 mutant CAR-T cells and expanding said plurality of IMPDH2 mutant CAR-T cells in the presence of mycophenolate for a second period of time thereby producing an expanded population of IMPDH2 CAR-T cells, wherein the second period of time is at least 14 days; and (iii) genetically altering said expanded population of IMPDH2 mutant CAR-T cells to inactivate a T-cell receptor (TCR) gene within said IMPDH2 mutant CAR-T cells thereby producing a plurality of TCR knockout IMPDH2 mutant CAR-T cells and expanding said plurality of TCR knockout IMPDH2 mutant CAR-T cells for a third period of time thereby producing an expanded population of mycophenolate resistant CAR-T cells, wherein the third period of time is at least 2 days.

[0005] In another aspect is provided a method of producing an expanded population of mycophenolate resistant CAR-T cells, the method including: (i) contacting a plurality of T cells with a first expression vector including a first nucleic acid sequence encoding a mutated IMPDH2 protein thereby producing a plurality of IMPDH2 mutant T cells, wherein said mutated IMPDH2 protein confers mycophenolate resistance to said plurality of IMPDH2 mutant T cells and expanding said plurality of IMPDH2 mutant T cells in the presence of mycophenolate for a first period of time thereby producing an expanded population of IMPDH2 T cells, wherein the first period of time is at least 2 days; (ii) contacting said expanded population of IMPDH2 mutant T cells with a second expression vector including a second nucleic acid encoding a chimeric antigen receptor (CAR) thereby producing a plurality of IMPDH2 mutant CAR-T cells and expanding said plurality of IMPDH2 mutant CAR-T cells for a second period of time thereby producing an expanded population of IMPDH2 CAR-T cells, wherein the second period of time is at least 2 days; and (iii) genetically altering said expanded population of IMPDH2 mutant CAR-T cells to inactivate a T-cell receptor (TCR) gene within said IMPDH2 mutant CAR-T cells thereby producing a plurality of TCR knockout IMPDH2 mutant CAR-T cells and expanding said plurality of TCR knockout IMPDH2 mutant CAR-T cells for a third period of time thereby producing an expanded population of mycophenolate resistant CAR-T cells, wherein the third period of time is at least 2 days.

[0006] In another aspect is provided a method of producing an expanded population of mycophenolate resistant CAR-T cells, the method including: (i) contacting a plurality of T cells withPATENT Attorney Docket No.048537-668001WO a first expression vector including a first nucleic acid sequence encoding a mutated IMPDH2 protein thereby producing a plurality of IMPDH2 mutant T cells, wherein said mutated IMPDH2 protein confers mycophenolate resistance to said plurality of IMPDH2 mutant T cells and expanding said plurality of IMPDH2 mutant T cells in the presence of mycophenolate for a first period of time thereby producing an expanded population of IMPDH2 T cells, wherein the first period of time is at least 2 days; (ii) genetically altering said expanded population of IMPDH2 mutant T cells to inactivate a T-cell receptor (TCR) gene within said IMPDH2 mutant T cells thereby producing a plurality of TCR knockout IMPDH2 mutant T cells and expanding said plurality of TCR knockout IMPDH2 mutant T cells for a second period of time thereby producing an expanded population of TCR knockout IMPDH2 mutant T cells, wherein the second period of time is at least 2 days; and (iii) contacting said expanded population of TCR knockout IMPDH2 mutant T cells with a second expression vector including a second nucleic acid encoding a chimeric antigen receptor (CAR) thereby producing a plurality of TCR knockout IMPDH2 mutant CAR-T cells and expanding said plurality of TCR knockout IMPDH2 mutant CAR-T cells for a third period of time thereby producing an expanded population of mycophenolate resistant CAR-T cells, wherein the third period of time is at least 2 days.

[0007] In another aspect is provided a method of producing an expanded population of mycophenolate resistant CAR-T cells, the method including: (i) genetically altering a plurality of T cells to inactivate a T-cell receptor (TCR) gene within said plurality of T cells thereby producing a plurality of TCR knockout T cells and expanding said plurality of TCR knockout T cells for a first period of time thereby producing an expanded population of TCR knockout T cells, wherein the first period of time is at least 2 days; (ii) contacting said expanded population of TCR knockout T cells with a first expression vector including a first nucleic acid encoding a mutated IMPDH2 protein thereby producing a plurality of TCR knockout IMPDH2 mutant T cells, wherein said mutated IMPDH2 protein confers mycophenolate resistance to said plurality of TCR knockout IMPDH2 mutant T cells and expanding said plurality of TCR knockout IMPDH2 mutant T cells in the presence of mycophenolate for a second period of time thereby producing an expanded population of TCR knockout IMPDH2 mutant T cells, wherein the second period of time is at least 2 days; and (iii) contacting said expanded population of TCR knockout IMPDH2 mutant T cells with a secondPATENT Attorney Docket No.048537-668001WO expression vector including a second nucleic acid sequence encoding a chimeric antigen receptor (CAR) thereby producing a plurality of TCR knockout IMPDH2 mutant CAR-T cells and expanding said plurality of TCR knockout IMPDH2 mutant CAR-T cells for a third period of time thereby producing an expanded population of mycophenolate resistant CAR-T cells, wherein the first period of time is at least 2 days.

[0008] In another aspect is provided a method of treating cancer in a subject in need thereof, the method including administering a therapeutically effective amount of an expanded population of mycophenolate resistant CAR-T cells produced by the methods provided herein including embodiments thereof or the pharmaceutical compositions provided herein including embodiments thereof.

[0009] In another aspect is provided a pharmaceutical composition including an expanded population of mycophenolate resistant CAR-T cells produced by the methods provided herein including embodiments thereof and a pharmaceutically acceptable excipient.

[0010] In another aspect is provided a method of treating cancer in a subject in need thereof, the method including administering to the subject a therapeutically effective amount of allogeneic CAR T-cells and an effective amount of an immune disrupting agent, wherein the allogenic CAR T-cells include a chimeric antigen receptor capable of binding to an anti-cancer antigen.

[0011] In another aspect is provided a method of expanding a CAR T-cell population derived from cord blood, the method including culturing the CAR T-cell population in the presence of an inhibitor of inosine 5’-monophosphate dehydrogenase (IMPDH). BRIEF DESCRIPTION OF THE DRAWINGS

[0012] FIG.1 shows the expansion of H2 transduced Jurkat cells cultured in mycophenolate. Jurkat cells were cultured in 96-well plates using a defined media supplemented with varying concentrations of mycophenolate and incubated and analyzed in an Incucyte live-cell analysis system over a 72h timeframe. The cultures were maintained at 37 °C and no additions were made to the cultures during this time frame. The mycophenolate concentrations are shown in ng / ml. NT are no transduced control cells incubated in 3.0 ng / ml mycophenolate.PATENT Attorney Docket No.048537-668001WO

[0013] FIG.2 shows the increased expression of RFP / H2 with greater culture time. Jurkat cells cultured in 2 mycophenolate concentrations over time express greater amounts of RFP documenting the selection of cells that have greater H2 expression. We expect that the relative resistance to mycophenolate will increase with time in culture and will greatly exceed concentrations associated with human immunosuppression at time of harvest. DETAILED DESCRIPTION DEFINITIONS

[0014] While various embodiments and aspects of the present invention are shown and described herein, it will be obvious to those skilled in the art that such embodiments and aspects are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention.

[0015] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in the application including, without limitation, patents, patent applications, articles, books, manuals, and treatises are hereby expressly incorporated by reference in their entirety for any purpose.

[0016] The abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts.

[0017] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art. See, e.g., Singleton et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY 2nd ed., J. Wiley & Sons (New York, NY 1994); Sambrook et al., MOLECULAR CLONING, A LABORATORY MANUAL, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989). Any methods, devices and materials similar or equivalent to those described herein can be used in the practice of this invention.PATENT Attorney Docket No.048537-668001WO The following definitions are provided to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure.

[0018] "Nucleic acid" refers to nucleotides (e.g., deoxyribonucleotides or ribonucleotides) and polymers thereof in either single-, double- or multiple-stranded form, or complements thereof; or nucleosides (e.g., deoxyribonucleosides or ribonucleosides). In embodiments, “nucleic acid” does not include nucleosides. The terms “polynucleotide,” “oligonucleotide,” “oligo” or the like refer, in the usual and customary sense, to a linear sequence of nucleotides. The term “nucleoside” refers, in the usual and customary sense, to a glycosylamine including a nucleobase and a five-carbon sugar (ribose or deoxyribose). Non limiting examples, of nucleosides include cytidine, uridine, adenosine, guanosine, thymidine and inosine. The term “nucleotide” refers, in the usual and customary sense, to a single unit of a polynucleotide, i.e., a monomer. Nucleotides can be ribonucleotides, deoxyribonucleotides, or modified versions thereof. Examples of polynucleotides contemplated herein include single and double stranded DNA, single and double stranded RNA, and hybrid molecules having mixtures of single and double stranded DNA and RNA. Examples of nucleic acid, e.g. polynucleotides contemplated herein include any types of RNA, e.g. mRNA, siRNA, miRNA, and guide RNA and any types of DNA, genomic DNA, plasmid DNA, and minicircle DNA, and any fragments thereof. The term “duplex” in the context of polynucleotides refers, in the usual and customary sense, to double strandedness. Nucleic acids can be linear or branched. For example, nucleic acids can be a linear chain of nucleotides or the nucleic acids can be branched, e.g., such that the nucleic acids comprise one or more arms or branches of nucleotides. Optionally, the branched nucleic acids are repetitively branched to form higher ordered structures such as dendrimers and the like.

[0019] Nucleic acids, including e.g., nucleic acids with a phosphothioate backbone, can include one or more reactive moieties. As used herein, the term reactive moiety includes any group capable of reacting with another molecule, e.g., a nucleic acid or polypeptide through covalent, non-covalent or other interactions. By way of example, the nucleic acid can include an amino acid reactive moiety that reacts with an amino acid on a protein or polypeptide through a covalent, non-covalent or other interaction.PATENT Attorney Docket No.048537-668001WO

[0020] The terms also encompass nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides. Examples of such analogs include, without limitation, phosphodiester derivatives including, e.g., phosphoramidate, phosphorodiamidate, phosphorothioate (also known as phosphothioate having double bonded sulfur replacing oxygen in the phosphate), phosphorodithioate, phosphonocarboxylic acids, phosphonocarboxylates, phosphonoacetic acid, phosphonoformic acid, methyl phosphonate, boron phosphonate, or O-methylphosphoroamidite linkages (see Eckstein, OLIGONUCLEOTIDES AND ANALOGUES: A PRACTICAL APPROACH, Oxford University Press) as well as modifications to the nucleotide bases such as in 5-methyl cytidine or pseudouridine.; and peptide nucleic acid backbones and linkages. Other analog nucleic acids include those with positive backbones; non-ionic backbones, modified sugars, and non-ribose backbones (e.g. phosphorodiamidate morpholino oligos or locked nucleic acids (LNA) as known in the art), including those described in U.S. Patent Nos.5,235,033 and 5,034,506, and Chapters 6 and 7, ASC Symposium Series 580, CARBOHYDRATE MODIFICATIONS IN ANTISENSE RESEARCH, Sanghui & Cook, eds. Nucleic acids containing one or more carbocyclic sugars are also included within one definition of nucleic acids. Modifications of the ribose- phosphate backbone may be done for a variety of reasons, e.g., to increase the stability and half-life of such molecules in physiological environments or as probes on a biochip. Mixtures of naturally occurring nucleic acids and analogs can be made; alternatively, mixtures of different nucleic acid analogs, and mixtures of naturally occurring nucleic acids and analogs may be made. In embodiments, the internucleotide linkages in DNA are phosphodiester, phosphodiester derivatives, or a combination of both.

[0021] Nucleic acids can include nonspecific sequences. As used herein, the term "nonspecific sequence" refers to a nucleic acid sequence that contains a series of residues that are not designed to be complementary to or are only partially complementary to any other nucleic acid sequence. By way of example, a nonspecific nucleic acid sequence is a sequence of nucleic acid residues that does not function as an inhibitory nucleic acid when contacted with a cell or organism.PATENT Attorney Docket No.048537-668001WO

[0022] A polynucleotide is typically composed of a specific sequence of four nucleotide bases: adenine (A); cytosine (C); guanine (G); and thymine (T) (uracil (U) for thymine (T) when the polynucleotide is RNA). Thus, the term “polynucleotide sequence” is the alphabetical representation of a polynucleotide molecule; alternatively, the term may be applied to the polynucleotide molecule itself. This alphabetical representation can be input into databases in a computer having a central processing unit and used for bioinformatics applications such as functional genomics and homology searching. Polynucleotides may optionally include one or more non-standard nucleotide(s), nucleotide analog(s) and / or modified nucleotides.

[0023] The term “complement,” as used herein, refers to a nucleotide (e.g., RNA or DNA) or a sequence of nucleotides capable of base pairing with a complementary nucleotide or sequence of nucleotides. As described herein and commonly known in the art the complementary (matching) nucleotide of adenosine is thymidine and the complementary (matching) nucleotide of guanosine is cytosine. Thus, a complement may include a sequence of nucleotides that base pair with corresponding complementary nucleotides of a second nucleic acid sequence. The nucleotides of a complement may partially or completely match the nucleotides of the second nucleic acid sequence. Where the nucleotides of the complement completely match each nucleotide of the second nucleic acid sequence, the complement forms base pairs with each nucleotide of the second nucleic acid sequence. Where the nucleotides of the complement partially match the nucleotides of the second nucleic acid sequence only some of the nucleotides of the complement form base pairs with nucleotides of the second nucleic acid sequence. Examples of complementary sequences include coding and a non-coding sequences, wherein the non-coding sequence contains complementary nucleotides to the coding sequence and thus forms the complement of the coding sequence. A further example of complementary sequences are sense and antisense sequences, wherein the sense sequence contains complementary nucleotides to the antisense sequence and thus forms the complement of the antisense sequence.

[0024] As described herein the complementarity of sequences may be partial, in which only some of the nucleic acids match according to base pairing, or complete, where all the nucleic acids match according to base pairing. Thus, two sequences that are complementary to each other, may have aPATENT Attorney Docket No.048537-668001WO specified percentage of nucleotides that are the same (i.e., about 60% identity, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region).

[0025] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, γ-carboxyglutamate, and O- phosphoserine. Amino acid analogs refers to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., an α carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid. The terms “non-naturally occurring amino acid” and “unnatural amino acid” refer to amino acid analogs, synthetic amino acids, and amino acid mimetics which are not found in nature.

[0026] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise, may be referred to by their commonly accepted single-letter codes.

[0027] The terms "polypeptide," "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues, wherein the polymer may In embodiments be conjugated to a moiety that does not consist of amino acids. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. A "fusion protein" refers to a chimeric protein encoding two or more separate protein sequences that are recombinantly expressed as a single moiety.PATENT Attorney Docket No.048537-668001WO

[0028] An amino acid or nucleotide base "position" is denoted by a number that sequentially identifies each amino acid (or nucleotide base) in the reference sequence based on its position relative to the N-terminus (or 5'-end). Due to deletions, insertions, truncations, fusions, and the like that must be considered when determining an optimal alignment, in general the amino acid residue number in a test sequence determined by simply counting from the N-terminus will not necessarily be the same as the number of its corresponding position in the reference sequence. For example, in a case where a variant has a deletion relative to an aligned reference sequence, there will be no amino acid in the variant that corresponds to a position in the reference sequence at the site of deletion. Where there is an insertion in an aligned reference sequence, that insertion will not correspond to a numbered amino acid position in the reference sequence. In the case of truncations or fusions there can be stretches of amino acids in either the reference or aligned sequence that do not correspond to any amino acid in the corresponding sequence.

[0029] The terms "numbered with reference to" or "corresponding to," when used in the context of the numbering of a given amino acid or polynucleotide sequence, refers to the numbering of the residues of a specified reference sequence when the given amino acid or polynucleotide sequence is compared to the reference sequence. An amino acid residue in a protein "corresponds" to a given residue when it occupies the same essential structural position within the protein as the given residue. One skilled in the art will immediately recognize the identity and location of residues corresponding to a specific position in a protein (e.g., CD19) in other proteins with different numbering systems. For example, by performing a simple sequence alignment with a protein (e.g., CD19) the identity and location of residues corresponding to specific positions of the protein are identified in other protein sequences aligning to the protein. For example, a selected residue in a selected protein corresponds to glutamic acid at position 138 when the selected residue occupies the same essential spatial or other structural relationship as a glutamic acid at position 138. In some embodiments, where a selected protein is aligned for maximum homology with a protein, the position in the aligned selected protein aligning with glutamic acid 138 is the to correspond to glutamic acid 138. Instead of a primary sequence alignment, a three dimensional structural alignment can also be used, e.g., where the structure of the selected protein is aligned for maximum correspondence with the glutamic acid at position 138, and the overall structures compared. In thisPATENT Attorney Docket No.048537-668001WO case, an amino acid that occupies the same essential position as glutamic acid 138 in the structural model is said to correspond to the glutamic acid 138 residue.

[0030] "Conservatively modified variants" applies to both amino acid and nucleic acid sequences. With respect to particular nucleic acid sequences, "conservatively modified variants" refers to those nucleic acids that encode identical or essentially identical amino acid sequences. Because of the degeneracy of the genetic code, a number of nucleic acid sequences will encode any given protein. For instance, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine. Thus, at every position where an alanine is specified by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are "silent variations," which are one species of conservatively modified variations. Every nucleic acid sequence herein which encodes a polypeptide also describes every possible silent variation of the nucleic acid. One of skill will recognize that each codon in a nucleic acid (except AUG, which is ordinarily the only codon for methionine, and TGG, which is ordinarily the only codon for tryptophan) can be modified to yield a functionally identical molecule. Accordingly, each silent variation of a nucleic acid which encodes a polypeptide is implicit in each described sequence.

[0031] As to amino acid sequences, one of skill will recognize that individual substitutions, deletions or additions to a nucleic acid, peptide, polypeptide, or protein sequence which alters, adds or deletes a single amino acid or a small percentage of amino acids in the encoded sequence is a "conservatively modified variant" where the alteration results in the substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants are in addition to and do not exclude polymorphic variants, interspecies homologs, and alleles of the disclosure.

[0032] The following eight groups each contain amino acids that are conservative substitutions for one another: 1) Alanine (A), Glycine (G); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine (Q);PATENT Attorney Docket No.048537-668001WO 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W); 7) Serine (S), Threonine (T); and 8) Cysteine (C), Methionine (M) (see, e.g., Creighton, Proteins (1984)).

[0033] The terms "identical" or percent "identity," in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., about 60% identity, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region, when compared and aligned for maximum correspondence over a comparison window or designated region) as measured using a BLAST or BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection (see, e.g., NCBI web site http: / / www.ncbi.nlm.nih.gov / BLAST / or the like). Such sequences are then said to be "substantially identical." This definition also refers to, or may be applied to, the compliment of a test sequence. The definition also includes sequences that have deletions and / or additions, as well as those that have substitutions. As described below, the preferred algorithms can account for gaps and the like. Preferably, identity exists over a region that is at least about 25 amino acids or nucleotides in length, or more preferably over a region that is 50- 100 amino acids or nucleotides in length.

[0034] "Percentage of sequence identity" is determined by comparing two optimally aligned sequences over a comparison window, wherein the portion of the polynucleotide or polypeptide sequence in the comparison window may comprise additions or deletions (i.e., gaps) as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number ofPATENT Attorney Docket No.048537-668001WO matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity.

[0035] A "comparison window", as used herein, includes reference to a segment of any one of the number of contiguous positions selected from the group consisting of, e.g., a full length sequence or from 20 to 600, about 50 to about 200, or about 100 to about 150 amino acids or nucleotides in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Methods of alignment of sequences for comparison are well-known in the art. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith and Waterman (1970) Adv. Appl. Math. 2:482c, by the homology alignment algorithm of Needleman and Wunsch (1970) J. Mol. Biol. 48:443, by the search for similarity method of Pearson and Lipman (1988) Proc. Nat’l. Acad. Sci. USA 85:2444, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by manual alignment and visual inspection (see, e.g., Ausubel et al., Current Protocols in Molecular Biology (1995 supplement)).

[0036] An example of an algorithm that is suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al. (1977) Nuc. Acids Res.25:3389-3402, and Altschul et al. (1990) J. Mol. Biol.215:403-410, respectively. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (http: / / www.ncbi.nlm.nih.gov / ). This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matchingPATENT Attorney Docket No.048537-668001WO residues; always > 0) and N (penalty score for mismatching residues; always < 0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a word length (W) of 11, an expectation (E) or 10, M=5, N=-4 and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a word length of 3, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff and Henikoff (1989) Proc. Natl. Acad. Sci. USA 89:10915) alignments (B) of 50, expectation (E) of 10, M=5, N=-4, and a comparison of both strands.

[0037] The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5787). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a reference sequence if the smallest sum probability in a comparison of the test nucleic acid to the reference nucleic acid is less than about 0.2, more preferably less than about 0.01, and most preferably less than about 0.001.

[0038] An indication that two nucleic acid sequences or polypeptides are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically cross reactive with the antibodies raised against the polypeptide encoded by the second nucleic acid, as described below. Thus, a polypeptide is typically substantially identical to a second polypeptide, for example, where the two peptides differ only by conservative substitutions. Another indication that two nucleic acid sequences are substantially identical is that the two molecules or their complements hybridize to each other under stringent conditions, as described below. Yet another indication that two nucleicPATENT Attorney Docket No.048537-668001WO acid sequences are substantially identical is that the same primers can be used to amplify the sequence.

[0039] The phrase "specifically (or selectively) binds" to an antibody or "specifically (or selectively) immunoreactive with," when referring to a protein or peptide, refers to a binding reaction that is determinative of the presence of the protein, often in a heterogeneous population of proteins and other biologics. Thus, under designated immunoassay conditions, the specified antibodies bind to a particular protein at least two times the background and more typically more than 10 to 100 times background. Specific binding to an antibody under such conditions requires an antibody that is selected for its specificity for a particular protein. For example, polyclonal antibodies can be selected to obtain only a subset of antibodies that are specifically immunoreactive with the selected antigen and not with other proteins. This selection may be achieved by subtracting out antibodies that cross-react with other molecules. A variety of immunoassay formats may be used to select antibodies specifically immunoreactive with a particular protein. For example, solid-phase ELISA immunoassays are routinely used to select antibodies specifically immunoreactive with a protein (see, e.g., Harlow & Lane, Using Antibodies, A Laboratory Manual (1998) for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity).

[0040] A "ligand" refers to an agent, e.g., a polypeptide or other molecule, capable of binding to a receptor or antibody, antibody variant, antibody region or fragment thereof.

[0041] Techniques for conjugating therapeutic agents to antibodies are well known (see, e.g., Arnon et al., "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy", in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp.243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., “Antibodies For Drug Delivery”in Controlled Drug Delivery (2ndEd.), Robinson et al. (eds.), pp.623-53 (Marcel Dekker, Inc.1987); Thorpe, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review" in Monoclonal Antibodies ‘84: Biological And Clinical Applications, Pinchera et al. (eds.), pp.475-506 (1985); and Thorpe et al., "The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates", Immunol. Rev., 62:119-58 (1982)). As used herein, the term “antibody-drug conjugate” or “ADC” refers to a therapeutic agent conjugated or otherwise covalently bound to to an antibody.PATENT Attorney Docket No.048537-668001WO

[0042] For specific proteins described herein, the named protein includes any of the protein’s naturally occurring forms, variants or homologs that maintain the protein transcription factor activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to the native protein). In some embodiments, variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g. a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring form. In other embodiments, the protein is the protein as identified by its NCBI sequence reference. In other embodiments, the protein is the protein as identified by its NCBI sequence reference, homolog or functional fragment thereof.

[0043] The term “ROR-1” or “ROR1” as used herein refers to any of the recombinant or naturally- occurring forms of tyrosine kinase-like orphan receptor 1 (ROR-1) or variants or homologs thereof that maintain ROR-1 activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to ROR-1). In some aspects, the variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring ROR-1 protein. In embodiments, the ROR-1 protein is substantially identical to the protein identified by Accession No. NP_005003.1 or a variant or homolog having substantial identity thereto. In embodiments, the ROR-1 protein is substantially identical to the protein identified by UniProt No. Q01973 or a variant or homolog having substantial identity thereto.

[0044] The term “CD19” or “Cluster of Differentiation 19” as used herein refers to any of the recombinant or naturally-occurring forms of the transmembrane protein B-lymphocyte antigen CD19, also known as B-lymphocyte surface antigen B4, or variants or homologs thereof that maintain CD19 activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to CD19). In some aspects, the variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring CD19 protein. In embodiments, the CD19 protein is substantially identical to the protein identified by UniProt No. P15391 or a variant or homolog having substantial identity thereto.PATENT Attorney Docket No.048537-668001WO

[0045] The term “BCMA” or “B-cell maturation antigen” as used herein refers to any of the recombinant or naturally-occurring forms of the cell surface receptor B-cell maturation antigen, also known as tumor necrosis factor receptor superfamily member 17 (TNFRSF17), or variants or homologs thereof that maintain BCMA activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to BCMA). In some aspects, the variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring BCMA protein. In embodiments, the BCMA protein is substantially identical to the protein identified by UniProt No. Q02223 or a variant or homolog having substantial identity thereto.

[0046] The term “CD123” or “Cluster of Differentiation 123” as used herein refers to any of the recombinant or naturally-occurring forms of interleukin-3 receptor, or variants or homologs thereof that maintain CD123 activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to CD123). In some aspects, the variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring CD123 protein. In embodiments, the CD123 protein is substantially identical to the protein identified by UniProt No. O75794 or a variant or homolog having substantial identity thereto.

[0047] The term “CS1” or “CD2 subset-1” as used herein refers to any of the recombinant or naturally-occurring forms of the surface antigen SLAM family member 7 (SLAMF7), also known as CD319, or variants or homologs thereof that maintain CS1 activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to CS1). In some aspects, the variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring CS1 protein. In embodiments, the CS1 protein is substantially identical to the protein identified by UniProt No. Q9NQ25 or a variant or homolog having substantial identity thereto.PATENT Attorney Docket No.048537-668001WO

[0048] The term “inosine 5’-monophosphate dehydrogenase” or “IMPDH” as used herein refers to any of the recombinant or naturally-occurring forms of the purine biosynthetic enzyme inosine 5’- monophosphate dehydrogenase or variants or homologs thereof that maintain IMPDH activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to IMPDH). In some aspects, the variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring IMPDH protein. In embodiments, the IMPDH protein is an IMPDH1 protein. In embodiments, the IMPDH1 protein is substantially identical to the protein identified by UniProt No. P20839 or a variant or homolog having substantial identity thereto. In embodiments, the IMPDH protein is an IMPDH2 protein. In embodiments, the IMPDH2 protein is critical for de novo guanine nucleotide synthesis in lymphocytes. In embodiments, the IMPDH2 protein is substantially identical to the protein identified by UniProt No. P12268 or a variant or homolog having substantial identity thereto. In embodiments, the IMPDH2 protein includes the amino acid sequence of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. In embodiments, the IMPDH2 protein includes the amino acid sequence of SEQ ID NO:1. In embodiments, the IMPDH2 protein includes the amino acid sequence of SEQ ID NO:2. In embodiments, the IMPDH2 protein includes the amino acid sequence of SEQ ID NO:3.

[0049] The term “mycophenolic acid” is used herein according to its plain ordinary meaning and refers to an immunosuppressant compound that inhibits de novo purine biosynthesis. In embodiments, the mycophenolic acid is an IMPDH inhibitor.

[0050] The term “mycophenolate” is used herein according to its plain ordinary meaning and refers to an immunosuppressant compound that inhibits de novo purine biosynthesis. In embodiments, the mycophenolate is an IMPDH inhibitor. In embodiments, the mycophenolate is an IMPDH2 inhibitor. In embodiments, the mycophenolate is mycophenolate mofetil (MMF) or mycophenolate sodium, or a pharmaceutically acceptable salt, ester, prodrug, or combinations thereof. In embodiments, the mycophenolate suppresses T cell proliferation and / or activation.

[0051] The term “mycophenolate resistant CAR-T cell” is used herein according to its plain and ordinary meaning and refers to a genetically modified T cell. In embodiments, mycophenolatePATENT Attorney Docket No.048537-668001WO resistant CAR-T cell expresses a chimeric antigen receptor (CAR) and exhibits resistance to mycophenolate. In embodiments, the resistance to mycophenolate is conferred by expression of a mutated inosine monophosphate dehydrogenase 2 (IMPDH2) protein. In embodiments, the mycophenolate resistant CAR-T cell is capable of proliferating and / or maintaining effector function in the presence of mycophenolate.

[0052] The term “chimeric antigen receptor protein” or “CAR protein” is used herein according to its plain and ordinary meaning and refers to a recombinant protein that includes both antigen binding and T cell activating functions. In embodiments, the CAR protein includes a heavy chain variable domain and a light chain variable domain which binds antigens. In embodiments, the binding of aCAR to an antigen activates a T cell. In embodiments, the CAR combines antigen-recognitiondomains with intracellular signaling domains to redirect T cell specificity and function. In embodiments, the CAR includes an extracellular antigen-binding domain (e.g., a single-chain variable fragment), a transmembrane domain, and one or more intracellular signaling domains (e.g., CD3ζ, CD28, 4-1BB). In embodiments, the CAR is specific for a cancer-associated antigen.

[0053] The term “CAR-T cell” is used herein according to its plain and ordinary meaning and refers to a T cell that has been genetically modified to express a chimeric antigen receptor (CAR). In embodiments, the CAR-T cell is capable of recognizing and binding to a target antigen expressed on a cancer cell, thereby mediating cytotoxic activity against the cancer cell. Methods for genetically modifying T cells are well known in the art (See Dimitri et al., Mol Cancer, 2022, 21, 78; Dai et al., Nat Methods, 2019, 16, p.247-54; Liu et al., Cell Res, 2017, 27, pp.154-57; Ren et al., Clin Cancer Res, 2017, 23, pp.2255-66; Rupp et al., Sci Rep, 2017, 7, 737; each of which is incorporated herein by reference in its entirety and for all purposes).

[0054] The term “IMPDH2 mutant CAR-T cell” is used herein according to its plain and ordinary meaning and refers to a CAR-T cell that expresses a mutated inosine monophosphate dehydrogenase 2 (IMPDH2) protein. In embodiments, the mutated IMPDH2 protein confers resistance to mycophenolate. In embodiments, the IMPDH2 mutant CAR-T cell is capable of proliferating and / or maintaining effector function in the presence of mycophenolate. In embodiments, the mutated IMPDH2 protein includes an amino acid having at least 80%, at least 81%, at least 82%, at leastPATENT Attorney Docket No.048537-668001WO 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 100% sequence identity to the sequence of SEQ ID NO:2.

[0055] The term “T cell receptor” or “TCR” is used herein according to its plain and ordinary meaning and refers to a protein expressed on the surface of a T cell that is responsible for antigen recognition. In embodiments, the TCR protein binds an antigen peptide bound to a major histocompatibility complex. In embodiments, the TCR protein is an alpha (α) chain, a beta (β) chain, a gamma (γ) chain, or a delta (δ) chain. In embodiments, the binding of an antigen peptide to the TCR protein results in the activation of the T cell. Methods for genetically modifying T cells to inactivate a TCR protein are well known in the art (See Liu et al., Cell Res, 2017, 27, pp.154-57; Ren et al., Clin Cancer Res, 2017, 23, pp.2255-66; Rupp et al., Sci Rep, 2017, 7, 737; each of which is incorporated herein by reference in its entirety and for all purposes).

[0056] The term “TCR knockout IMPDH2 mutant CAR-T cell” refers to an IMPDH2 mutant CAR-T cell in which one or more genes encoding the T-cell receptor (TCR) have been genetically inactivated. In embodiments, the TCR knockout is achieved using genome editing technologies such as CRISPR or TALEN.

[0057] The term "gene" means the segment of DNA involved in producing a protein; it includes regions preceding and following the coding region (leader and trailer) as well as intervening sequences (introns) between individual coding segments (exons). The leader, the trailer as well as the introns include regulatory elements that are necessary during the transcription and the translation of a gene. Further, a "protein gene product" is a protein expressed from a particular gene.

[0058] The terms "plasmid", "vector" or "expression vector" refer to a nucleic acid molecule that encodes for genes and / or regulatory elements necessary for the expression of genes. Expression of a gene from a plasmid can occur in cis or in trans. If a gene is expressed in cis, the gene and the regulatory elements are encoded by the same plasmid. Expression in trans refers to the instance where the gene and the regulatory elements are encoded by separate plasmids.PATENT Attorney Docket No.048537-668001WO

[0059] The word "expression" or "expressed" as used herein in reference to a gene means the transcriptional and / or translational product of that gene. The level of expression of a DNA molecule in a cell may be determined on the basis of either the amount of corresponding mRNA that is present within the cell or the amount of protein encoded by that DNA produced by the cell. The level of expression of non-coding nucleic acid molecules may be detected by standard PCR or Northern blot methods well known in the art. See, Sambrook et al., 1989 Molecular Cloning: A Laboratory Manual, 18.1-18.88.

[0060] Expression of a transfected gene can occur transiently or stably in a cell. During "transient expression" the transfected gene is not transferred to the daughter cell during cell division. Since its expression is restricted to the transfected cell, expression of the gene is lost over time. In contrast, stable expression of a transfected gene can occur when the gene is co-transfected with another gene that confers a selection advantage to the transfected cell. Such a selection advantage may be a resistance towards a certain toxin that is presented to the cell.

[0061] The terms "transfection", "transduction", "transfecting" or "transducing" can be used interchangeably and are defined as a process of introducing a nucleic acid molecule or a protein to a cell. Nucleic acids are introduced to a cell using non-viral or viral-based methods. The nucleic acid molecules may be gene sequences encoding complete proteins or functional portions thereof. Non- viral methods of transfection include any appropriate transfection method that does not use viral DNA or viral particles as a delivery system to introduce the nucleic acid molecule into the cell. Exemplary non-viral transfection methods include calcium phosphate transfection, liposomal transfection, nucleofection, sonoporation, transfection through heat shock, magnetifection and electroporation. In some embodiments, the nucleic acid molecules are introduced into a cell using electroporation following standard procedures well known in the art. For viral-based methods of transfection any useful viral vector may be used in the methods described herein. Examples for viral vectors include, but are not limited to retroviral, adenoviral, lentiviral and adeno-associated viral vectors. In some embodiments, the nucleic acid molecules are introduced into a cell using a retroviral vector following standard procedures well known in the art. The terms ″transfection″ or ″transduction″ also refer to introducing proteins into a cell from the external environment. Typically,PATENT Attorney Docket No.048537-668001WO transduction or transfection of a protein relies on attachment of a peptide or protein capable of crossing the cell membrane to the protein of interest. See, e.g., Ford et al. (2001) Gene Therapy 8:1- 4 and Prochiantz (2007) Nat. Methods 4:119-20.

[0062] A "label" or a "detectable moiety" is a composition detectable by spectroscopic, photochemical, biochemical, immunochemical, chemical, or other physical means. For example, useful labels include 32P, fluorescent dyes, electron-dense reagents, enzymes (e.g., as commonly used in an ELISA), biotin, digoxigenin, or haptens and proteins or other entities which can be made detectable, e.g., by incorporating a radiolabel into a peptide or antibody specifically reactive with a target peptide. Any appropriate method known in the art for conjugating an antibody to the label may be employed, e.g., using methods described in Hermanson, Bioconjugate Techniques 1996, Academic Press, Inc., San Diego.

[0063] When the label or detectable moiety is a radioactive metal or paramagnetic ion, the agent may be reacted with another long-tailed reagent having a long tail with one or more chelating groups attached to the long tail for binding to these ions. The long tail may be a polymer such as a polylysine, polysaccharide, or other derivatized or derivatizable chain having pendant groups to which the metals or ions may be added for binding. Examples of chelating groups that may be used according to the disclosure include, but are not limited to, ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), DOTA, NOTA, NETA, TETA, porphyrins, polyamines, crown ethers, bis-thiosemicarbazones, polyoximes, and like groups. The chelate is normally linked to the PSMA antibody or functional antibody fragment by a group, which enables the formation of a bond to the molecule with minimal loss of immunoreactivity and minimal aggregation and / or internal cross-linking. The same chelates, when complexed with non-radioactive metals, such as manganese, iron and gadolinium are useful for MRI, when used along with the antibodies and carriers described herein. Macrocyclic chelates such as NOTA, DOTA, and TETA are of use with a variety of metals and radiometals including, but not limited to, radionuclides of gallium, yttrium and copper, respectively. Other ring-type chelates such as macrocyclic polyethers, which are of interest for stably binding nuclides, such as223Ra for RAIT may be used. In certainPATENT Attorney Docket No.048537-668001WO embodiments, chelating moieties may be used to attach a PET imaging agent, such as an Al-18F complex, to a targeting molecule for use in PET analysis.

[0064] The term "antigen" as provided herein refers to molecules capable of binding to the antibody binding domain provided herein. An "antigen binding domain" as provided herein is a region of an antibody that binds to an antigen (epitope). As described above, the antigen binding domain is generally composed of one constant and one variable domain of each of the heavy and the light chain (VL, VH, CLand CH1, respectively). The paratope or antigen-binding site is formed on the N-terminus of the antigen binding domain. The two variable domains of an antigen binding domain typically bind the epitope on an antigen.

[0065] "Contacting" is used in accordance with its plain ordinary meaning and refers to the process of allowing at least two distinct species (e.g. antibodies and antigens) to become sufficiently proximal to react, interact, or physically touch. It should be appreciated, however, that the resulting reaction product can be produced directly from a reaction between the added reagents or from an intermediate from one or more of the added reagents which can be produced in the reaction mixture.

[0066] The term "contacting" may include allowing two species to react, interact, or physically touch, wherein the two species may be, for example, a pharmaceutical composition as provided herein and a cell. In embodiments contacting includes, for example, allowing a pharmaceutical composition as described herein to interact with a cell.

[0067] A "cell" as used herein, refers to a cell carrying out metabolic or other function sufficient to preserve or replicate its genomic DNA. A cell can be identified by well-known methods in the art including, for example, presence of an intact membrane, staining by a particular dye, ability to produce progeny or, in the case of a gamete, ability to combine with a second gamete to produce a viable offspring. Cells may include prokaryotic and eukaryotic cells. Prokaryotic cells include but are not limited to bacteria. Eukaryotic cells include, but are not limited to, yeast cells and cells derived from plants and animals, for example mammalian, insect (e.g., spodoptera) and human cells.

[0068] The term "recombinant" when used with reference, e.g., to a cell, nucleic acid, protein, or vector, indicates that the cell, nucleic acid, protein or vector, has been modified by the introductionPATENT Attorney Docket No.048537-668001WO of a heterologous nucleic acid or protein or the alteration of a native nucleic acid or protein, or that the cell is derived from a cell so modified. Thus, for example, recombinant cells express genes that are not found within the native (non-recombinant) form of the cell or express native genes that are otherwise abnormally expressed, under expressed or not expressed at all. Transgenic cells and plants are those that express a heterologous gene or coding sequence, typically as a result of recombinant methods.

[0069] The term "isolated", when applied to a nucleic acid or protein, denotes that the nucleic acid or protein is essentially free of other cellular components with which it is associated in the natural state. It can be, for example, in a homogeneous state and may be in either a dry or aqueous solution. Purity and homogeneity are typically determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high performance liquid chromatography. A protein that is the predominant species present in a preparation is substantially purified.

[0070] The term "heterologous" when used with reference to portions of a nucleic acid indicates that the nucleic acid comprises two or more subsequences that are not found in the same relationship to each other in nature. For instance, the nucleic acid is typically recombinantly produced, having two or more sequences from unrelated genes arranged to make a new functional nucleic acid, e.g., a promoter from one source and a coding region from another source. Similarly, a heterologous protein indicates that the protein comprises two or more subsequences that are not found in the same relationship to each other in nature (e.g., a fusion protein).

[0071] The term "exogenous" refers to a molecule or substance (e.g., a compound, nucleic acid or protein) that originates from outside a given cell or organism. For example, an "exogenous promoter" as referred to herein is a promoter that does not originate from the cell or organism it is expressed by. Conversely, the term "endogenous" or "endogenous promoter" refers to a molecule or substance that is native to, or originates within, a given cell or organism.

[0072] As defined herein, the term "inhibition", "inhibit", "inhibiting" and the like in reference to cell proliferation (e.g., cancer cell proliferation) means negatively affecting (e.g., decreasing proliferation) or killing the cell. In some embodiments, inhibition refers to reduction of a disease orPATENT Attorney Docket No.048537-668001WO symptoms of disease (e.g., cancer, cancer cell proliferation). Thus, inhibition includes, at least in part, partially or totally blocking stimulation, decreasing, preventing, or delaying activation, or inactivating, desensitizing, or down-regulating signal transduction or enzymatic activity or the amount of a protein (e.g. a cancer-associated protein or IMPDH). In embodiments inhibition refers to reduction of a disease or symptoms of disease. In embodiments, inhibition refers to a reduction in the activity of a particular protein target (e.g., IMPDH). Thus, inhibition includes, at least in part, partially or totally blocking stimulation, decreasing, preventing, or delaying activation, or inactivating, desensitizing, or down-regulating signal transduction or enzymatic activity or the amount of a protein (e.g., IMPDH). In embodiments, inhibition refers to a reduction of activity of a target protein resulting from a direct interaction (e.g. an inhibitor binds to the target protein). In embodiments, inhibition refers to a reduction of activity of a target protein from an indirect interaction (e.g. an inhibitor binds to a protein that activates the target protein, thereby preventing target protein activation). Similarly, an "inhibitor" is a compound or protein that inhibits a receptor or another protein, e.g.,, by binding, partially or totally blocking, decreasing, preventing, delaying, inactivating, desensitizing, or down-regulating activity (e.g., a receptor activity or a protein activity).

[0073] The term "expression" includes any step involved in the production of the polypeptide including, but not limited to, transcription, post-transcriptional modification, translation, post- translational modification, and secretion. Expression can be detected using conventional techniques for detecting protein (e.g., ELISA, Western blotting, flow cytometry, immunofluorescence, immunohistochemistry, etc.).

[0074] “Biological sample” or “sample” refer to materials obtained from or derived from a subject or patient. A biological sample includes sections of tissues such as biopsy and autopsy samples, and frozen sections taken for histological purposes. Such samples include bodily fluids such as blood and blood fractions or products (e.g., serum, plasma, platelets, red blood cells, and the like), sputum, tissue, cultured cells (e.g., primary cultures, explants, and transformed cells) stool, urine, synovial fluid, joint tissue, synovial tissue, synoviocytes, fibroblast-like synoviocytes, macrophage-like synoviocytes, immune cells, hematopoietic cells, fibroblasts, macrophages, T cells, etc. A biological sample is typically obtained from a eukaryotic organism, such as a mammal such as a primate e.g.,PATENT Attorney Docket No.048537-668001WO chimpanzee or human; cow; dog; cat; a rodent, e.g., guinea pig, rat, mouse; rabbit; or a bird; reptile; or fish.

[0075] A “control” or “standard control” refers to a sample, measurement, or value that serves as a reference, usually a known reference, for comparison to a test sample, measurement, or value. For example, a test sample can be taken from a patient suspected of having a given disease (e.g. cancer) and compared to a known normal (non-diseased) individual (e.g. a standard control subject). A standard control can also represent an average measurement or value gathered from a population of similar individuals (e.g. standard control subjects) that do not have a given disease (i.e. standard control population), e.g., healthy individuals with a similar medical background, same age, weight, etc. A standard control value can also be obtained from the same individual, e.g. from an earlier- obtained sample from the patient prior to disease onset. For example, a control can be devised to compare therapeutic benefit based on pharmacological data (e.g., half-life) or therapeutic measures (e.g., comparison of side effects). Controls are also valuable for determining the significance of data. For example, if values for a given parameter are widely variant in controls, variation in test samples will not be considered as significant. One of skill will recognize that standard controls can be designed for assessment of any number of parameters (e.g. RNA levels, protein levels, specific cell types, specific bodily fluids, specific tissues, etc).

[0076] One of skill in the art will understand which standard controls are most appropriate in a given situation and be able to analyze data based on comparisons to standard control values. Standard controls are also valuable for determining the significance (e.g. statistical significance) of data. For example, if values for a given parameter are widely variant in standard controls, variation in test samples will not be considered as significant.

[0077] As used herein, the term "cancer" or “tumor” refers to all types of cancer, neoplasm or malignant tumors found in mammals (e.g. humans), including leukemias, lymphomas, carcinomas and sarcomas. Exemplary cancers that may be treated with a compound or method provided herein include brain cancer, glioma, glioblastoma, neuroblastoma, prostate cancer, colorectal cancer, pancreatic cancer, Medulloblastoma, melanoma, cervical cancer, gastric cancer, ovarian cancer, lung cancer, cancer of the head, Hodgkin's Disease, and Non-Hodgkin's Lymphomas. Exemplary cancersPATENT Attorney Docket No.048537-668001WO that may be treated with a compound or method provided herein include cancer of the thyroid, endocrine system, brain, breast, cervix, colon, head & neck, liver, kidney, lung, ovary, pancreas, rectum, stomach, and uterus. Additional examples include, thyroid carcinoma, cholangiocarcinoma, pancreatic adenocarcinoma, skin cutaneous melanoma, colon adenocarcinoma, rectum adenocarcinoma, stomach adenocarcinoma, esophageal carcinoma, head and neck squamous cell carcinoma, breast invasive carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, non- small cell lung carcinoma, mesothelioma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia, primary brain tumors, malignant pancreatic insulanoma, malignant carcinoid, urinary bladder cancer, premalignant skin lesions, testicular cancer, thyroid cancer, neuroblastoma, esophageal cancer, genitourinary tract cancer, malignant hypercalcemia, endometrial cancer, adrenal cortical cancer, neoplasms of the endocrine or exocrine pancreas, medullary thyroid cancer, medullary thyroid carcinoma, melanoma, colorectal cancer, papillary thyroid cancer, hepatocellular carcinoma, or prostate cancer.

[0078] “Treating” or “treatment” as used herein (and as well-understood in the art) also broadly includes any approach for obtaining beneficial or desired results in a subject’s condition, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of the extent of a disease, stabilizing (i.e., not worsening) the state of disease, prevention of a disease’s transmission or spread, delay or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission, whether partial or total and whether detectable or undetectable. In other words, "treatment" as used herein includes any cure, amelioration, or prevention of a disease. Treatment may prevent the disease from occurring; inhibit the disease’s spread; relieve the disease’s symptoms.,, fully or partially remove the disease’s underlying cause, shorten a disease’s duration, or do a combination of these things.

[0079] "Treating" and "treatment" as used herein include prophylactic treatment. Treatment methods include administering to a subject a therapeutically effective amount of an active agent. The administering step may consist of a single administration or may include a series ofPATENT Attorney Docket No.048537-668001WO administrations. The length of the treatment period depends on a variety of factors, such as the severity of the condition, the age of the patient, the concentration of active agent, the activity of the compositions used in the treatment, or a combination thereof. It will also be appreciated that the effective dosage of an agent used for the treatment or prophylaxis may increase or decrease over the course of a particular treatment or prophylaxis regime. Changes in dosage may result and become apparent by standard diagnostic assays known in the art. In some instances, chronic administration may be required. For example, the compositions are administered to the subject in an amount and for a duration sufficient to treat the patient. In embodiments, the treating or treatment is no prophylactic treatment.

[0080] “Patient” or “subject in need thereof” refers to a living organism suffering from or prone to a disease or condition that can be treated by administration of a pharmaceutical composition as provided herein. Non-limiting examples include humans, other mammals, bovines, rats, mice, dogs, monkeys, goat, sheep, cows, deer, and other non-mammalian animals. In some embodiments, a patient is human.

[0081] A “effective amount” is an amount sufficient for a compound to accomplish a stated purpose relative to the absence of the compound (e.g., achieve the effect for which it is administered, treat a disease, reduce enzyme activity, increase enzyme activity, reduce a signaling pathway, or reduce one or more symptoms of a disease or condition). An example of an “effective amount” is an amount sufficient to contribute to the treatment, prevention, or reduction of a symptom or symptoms of a disease, which could also be referred to as a “therapeutically effective amount.” A “reduction” of a symptom or symptoms (and grammatical equivalents of this phrase) means decreasing of the severity or frequency of the symptom(s), or elimination of the symptom(s). A “prophylactically effective amount” of a drug is an amount of a drug that, when administered to a subject, will have the intended prophylactic effect, e.g., preventing or delaying the onset (or reoccurrence) of an injury, disease, pathology or condition, or reducing the likelihood of the onset (or reoccurrence) of an injury, disease, pathology, or condition, or their symptoms. The full prophylactic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses. Thus, a prophylactically effective amount may bePATENT Attorney Docket No.048537-668001WO administered in one or more administrations. An “activity decreasing amount,” as used herein, refers to an amount of antagonist required to decrease the activity of an enzyme relative to the absence of the antagonist. A “function disrupting amount,” as used herein, refers to the amount of antagonist required to disrupt the function of an enzyme or protein relative to the absence of the antagonist. The exact amounts will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols.1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

[0082] For any compound described herein, the therapeutically effective amount can be initially determined from cell culture assays. Target concentrations will be those concentrations of active compound(s) that are capable of achieving the methods described herein, as measured using the methods described herein or known in the art.

[0083] As is well known in the art, therapeutically effective amounts for use in humans can also be determined from animal models. For example, a dose for humans can be formulated to achieve a concentration that has been found to be effective in animals. The dosage in humans can be adjusted by monitoring compounds effectiveness and adjusting the dosage upwards or downwards, as described above. Adjusting the dose to achieve maximal efficacy in humans based on the methods described above and other methods is well within the capabilities of the ordinarily skilled artisan.

[0084] The term “therapeutically effective amount,” as used herein, refers to that amount of the therapeutic agent sufficient to ameliorate the disorder, as described above. For example, for the given parameter, a therapeutically effective amount will show an increase or decrease of at least 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90%, or at least 100%. Therapeutic efficacy can also be expressed as “-fold” increase or decrease. For example, a therapeutically effective amount can have at least a 1.2-fold, 1.5-fold, 2-fold, 5-fold, or more effect over a control.

[0085] Dosages may be varied depending upon the requirements of the patient and the compound being employed. The dose administered to a patient, in the context of the present disclosure, shouldPATENT Attorney Docket No.048537-668001WO be sufficient to effect a beneficial therapeutic response in the patient over time. The size of the dose also will be determined by the existence, nature, and extent of any adverse side-effects. Determination of the proper dosage for a particular situation is within the skill of the practitioner. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under circumstances is reached. Dosage amounts and intervals can be adjusted individually to provide levels of the administered compound effective for the particular clinical indication being treated. This will provide a therapeutic regimen that is commensurate with the severity of the individual's disease state.

[0086] As used herein, the term "administering" means oral administration, administration as a suppository, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal or subcutaneous administration, or the implantation of a slow-release device, e.g., a mini-osmotic pump, to a subject. Administration is by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, e.g., intravenous, intramuscular, intra-arteriole, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, etc. In embodiments, the administering does not include administration of any active agent other than the recited active agent.

[0087] "Co-administer" it is meant that a composition described herein is administered at the same time, just prior to, or just after the administration of one or more additional therapies. The compounds provided herein can be administered alone or can be coadministered to the patient. Coadministration is meant to include simultaneous or sequential administration of the compounds individually or in combination (more than one compound). Thus, the preparations can also be combined, when desired, with other active substances (e.g. to reduce metabolic degradation). The compositions of the present disclosure can be delivered transdermally, by a topical route, or formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols.PATENT Attorney Docket No.048537-668001WO

[0088] An “anticancer agent” as used herein refers to a molecule (e.g. compound, peptide, protein, nucleic acid, 0103) used to treat cancer through destruction or inhibition of cancer cells or tissues. Anticancer agents may be selective for certain cancers or certain tissues. In embodiments, anticanceragents herein may include epigenetic inhibitors and multi-kinase inhibit “Anti-cancer agent” and“anticancer agent” are used in accordance with their plain ordinary meaning and refers to a composition (e.g. compound, drug, antagonist, inhibitor, modulator) having antineoplastic properties or the ability to inhibit the growth or proliferation of cells. In some embodiments, an anti-cancer agent is a chemotherapeutic. In some embodiments, an anti-cancer agent is an agent identified herein having utility in methods of treating cancer. In some embodiments, an anti-cancer agent is an agent approved by the FDA or similar regulatory agency of a country other than the USA, for treating cancer. Examples of anti-cancer agents include, but are not limited to, MEK (e.g. MEK1, MEK2, or MEK1 and MEK2) inhibitors (e.g. XL518, CI-1040, PD035901, selumetinib / AZD6244, GSK1120212 / trametinib, GDC-0973, ARRY-162, ARRY-300, AZD8330, PD0325901, U0126, PD98059, TAK-733, PD318088, AS703026, BAY 869766), alkylating agents (e.g., cyclophosphamide, ifosfamide, chlorambucil, busulfan, melphalan, mechlorethamine, uramustine, thiotepa, nitrosoureas, nitrogen mustards (e.g., mechloroethamine, cyclophosphamide, chlorambucil, meiphalan), ethylenimine and methylmelamines (e.g., hexamethlymelamine, thiotepa), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine, lomusitne, semustine, streptozocin), triazenes (decarbazine)), anti-metabolites (e.g., 5- azathioprine, leucovorin, capecitabine, fludarabine, gemcitabine, pemetrexed, raltitrexed, folic acid analog (e.g., methotrexate), or pyrimidine analogs (e.g., fluorouracil, floxouridine, Cytarabine), purine analogs (e.g., mercaptopurine, thioguanine, pentostatin), etc.), plant alkaloids (e.g., vincristine, vinblastine, vinorelbine, vindesine, podophyllotoxin, paclitaxel, docetaxel, etc.), topoisomerase inhibitors (e.g., irinotecan, topotecan, amsacrine, etoposide (VP16), etoposide phosphate, teniposide, etc.), antitumor antibiotics (e.g., doxorubicin, adriamycin, daunorubicin, epirubicin, actinomycin, bleomycin, mitomycin, mitoxantrone, plicamycin, etc.), platinum-based compounds (e.g. cisplatin, oxaloplatin, carboplatin), anthracenedione (e.g., mitoxantrone), substituted urea (e.g., hydroxyurea), methyl hydrazine derivative (e.g., procarbazine), adrenocortical suppressant (e.g., mitotane, aminoglutethimide), epipodophyllotoxins (e.g., etoposide), antibiotics (e.g., daunorubicin,PATENT Attorney Docket No.048537-668001WO doxorubicin, bleomycin), enzymes (e.g., L-asparaginase), inhibitors of mitogen-activated protein kinase signaling (e.g. U0126, PD98059, PD184352, PD0325901, ARRY-142886, SB239063, SP600125, BAY 43-9006, wortmannin, or LY294002, Syk inhibitors, mTOR inhibitors, antibodies (e.g., rituxan), gossyphol, genasense, polyphenol E, Chlorofusin, all trans-retinoic acid (ATRA), bryostatin, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), 5-aza-2'- deoxycytidine, all trans retinoic acid, doxorubicin, vincristine, etoposide, gemcitabine, imatinib (Gleevec.RTM.), geldanamycin, 17-N-Allylamino-17-Demethoxygeldanamycin (17-AAG), flavopiridol, LY294002, bortezomib, trastuzumab, BAY 11-7082, PKC412, PD184352, 20-epi-1, 25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine; amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic protein-1; antiandrogen, prostatic carcinoma; antiestrogen; antineoplaston; antisense oligonucleotides; aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators; apurinic acid; ara-CDP-DL- PTBA; arginine deaminase; asulacrine; atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat; BCR / ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine; calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2; capecitabine; carboxamide- amino-triazole; carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine; clomifene analogues; clotrimazole; collismycin A; collismycin B; combretastatin A4; combretastatin analogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A; cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone; didemnin B; didox; diethylnorspermine; dihydro-5- azacytidine; 9-dioxamycin; diphenyl spiromustine; docosanol; dolasetron; doxifluridine;PATENT Attorney Docket No.048537-668001WO droloxifene; dronabinol; duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur; epirubicin; epristeride; estramustine analogue; estrogen agonists; estrogen antagonists; etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinide; filgrastim; finasteride; flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex; formestane; fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix; gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam; heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene; idramantone; ilmofosine; ilomastat; imidazoacridones; imiquimod; immunostimulant peptides; insulin-like growth factor-1 receptor inhibitor; interferon agonists; interferons; interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron; jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide; leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole; leukemia inhibiting factor; leukocyte alpha interferon; leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole; linear polyamine analogue; lipophilic disaccharide peptide; lipophilic platinum compounds; lissoclinamide 7; lobaplatin; lombricine; lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine; lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysin inhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone; meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone; mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonal antibody, human chorionic gonadotrophin; monophosphoryl lipid A+myobacterium cell wall sk; mopidamol; multiple drug resistance gene inhibitor; multiple tumor suppressor 1-based therapy; mustard anticancer agent; mycaperoxide B; mycobacterial cell wall extract; myriaporone; N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin; nemorubicin; neridronic acid; neutral endopeptidase; nilutamide; nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn; O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone; ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin; osaterone; oxaliplatin; oxaunomycin; palauamine; palmitoylrhizoxin; pamidronic acid; panaxytriol; panomifene;PATENT Attorney Docket No.048537-668001WO parabactin; pazelliptine; pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin; pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin; phenylacetate; phosphatase inhibitors; picibanil; pilocarpine hydrochloride; pirarubicin; piritrexim; placetin A; placetin B; plasminogen activator inhibitor; platinum complex; platinum compounds; platinum-triamine complex; porfimer sodium; porfiromycin; prednisone; propyl bis-acridone; prostaglandin J2; proteasome inhibitors; protein A-based immune modulator; protein kinase C inhibitor; protein kinase C inhibitors, microalgal; protein tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine; pyridoxylated hemoglobin polyoxyethylerie conjugate; raf antagonists; raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors; ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re 186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide; rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol; saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine; senescence derived inhibitor 1; sense oligonucleotides; signal transduction inhibitors; signal transduction modulators; single chain antigen-binding protein; sizofuran; sobuzoxane; sodium borocaptate; sodium phenylacetate; solverol; somatomedin binding protein; sonermin; sparfosic acid; spicamycin D; spiromustine; splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-cell division inhibitors; stipiamide; stromelysin inhibitors; sulfinosine; superactive vasoactive intestinal peptide antagonist; suradista; suramin; swainsonine; synthetic glycosaminoglycans; tallimustine; tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomerase inhibitors; temoporfin; temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine; thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic; thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocene bichloride; topsentin; toremifene; totipotent stem cell factor; translation inhibitors; tretinoin; triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenital sinus-derived growth inhibitory factor; urokinase receptor antagonists; vapreotide; variolin B; vector system, erythrocyte gene therapy; velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; zinostatin stimalamer, Adriamycin, Dactinomycin, Bleomycin, Vinblastine, Cisplatin, acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin;PATENT Attorney Docket No.048537-668001WO altretamine; ambomycin; ametantrone acetate; aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa; bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate; brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone; caracemide; carbetimer; carboplatin; carmustine; carubicin hydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin; cladribine; crisnatol mesylate; cyclophosphamide; cytarabine; dacarbazine; daunorubicin hydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin; edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine; epirubicin hydrochloride; erbulozole; esorubicin hydrochloride; estramustine; estramustine phosphate sodium; etanidazole; etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine; fenretinide; floxuridine; fludarabine phosphate; fluorouracil; fluorocitabine; fosquidone; fostriecin sodium; gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide; iimofosine; interleukin I1 (including recombinant interleukin II, or rlL.sub.2), interferon alfa-2a; interferon alfa-2b; interferon alfa-n1; interferon alfa-n3; interferon beta-1a; interferon gamma-1b; iproplatin; irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolide acetate; liarozole hydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride; masoprocol; maytansine; mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate; melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazoie; nogalamycin; ormaplatin; oxisuran; pegaspargase; peliomycin; pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine; procarbazine hydrochloride; puromycin; puromycin hydrochloride; pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride; semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone; testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifene citrate;PATENT Attorney Docket No.048537-668001WO trestolone acetate; triciribine phosphate; trimetrexate; trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracil mustard; uredepa; vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate; vindesine; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate; vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate; vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicin hydrochloride, agents that arrest cells in the G2-M phases and / or modulate the formation or stability of microtubules, (e.g. Taxol.TM (i.e. paclitaxel), Taxotere.TM, compounds comprising the taxane skeleton, Erbulozole (i.e. R-55104), Dolastatin 10 (i.e. DLS-10 and NSC-376128), Mivobulin isethionate (i.e. as CI-980), Vincristine, NSC-639829, Discodermolide (i.e. as NVP-XX- A-296), ABT-751 (Abbott, i.e. E-7010), Altorhyrtins (e.g. Altorhyrtin A and Altorhyrtin C), Spongistatins (e.g. Spongistatin 1, Spongistatin 2, Spongistatin 3, Spongistatin 4, Spongistatin 5, Spongistatin 6, Spongistatin 7, Spongistatin 8, and Spongistatin 9), Cemadotin hydrochloride (i.e. LU-103793 and NSC-D-669356), Epothilones (e.g. Epothilone A, Epothilone B, Epothilone C (i.e. desoxyepothilone A or dEpoA), Epothilone D (i.e. KOS-862, dEpoB, and desoxyepothilone B), Epothilone E, Epothilone F, Epothilone B N-oxide, Epothilone A N-oxide, 16-aza-epothilone B, 21- aminoepothilone B (i.e. BMS-310705), 21-hydroxyepothilone D (i.e. Desoxyepothilone F and dEpoF), 26-fluoroepothilone, Auristatin PE (i.e. NSC-654663), Soblidotin (i.e. TZT-1027), LS- 4559-P (Pharmacia, i.e. LS-4577), LS-4578 (Pharmacia, i.e. LS-477-P), LS-4477 (Pharmacia), LS- 4559 (Pharmacia), RPR-112378 (Aventis), Vincristine sulfate, DZ-3358 (Daiichi), FR-182877 (Fujisawa, i.e. WS-9885B), GS-164 (Takeda), GS-198 (Takeda), KAR-2 (Hungarian Academy of Sciences), BSF-223651 (BASF, i.e. ILX-651 and LU-223651), SAH-49960 (Lilly / Novartis), SDZ- 268970 (Lilly / Novartis), AM-97 (Armad / Kyowa Hakko), AM-132 (Armad), AM-138 (Armad / Kyowa Hakko), IDN-5005 (Indena), Cryptophycin 52 (i.e. LY-355703), AC-7739 (Ajinomoto, i.e. AVE-8063A and CS-39.HCl), AC-7700 (Ajinomoto, i.e. AVE-8062, AVE-8062A, CS-39-L-Ser.HCl, and RPR-258062A), Vitilevuamide, Tubulysin A, Canadensol, Centaureidin (i.e. NSC-106969), T-138067 (Tularik, i.e. T-67, TL-138067 and TI-138067), COBRA-1 (Parker Hughes Institute, i.e. DDE-261 and WHI-261), H10 (Kansas State University), H16 (Kansas State University), Oncocidin A1 (i.e. BTO-956 and DIME), DDE-313 (Parker Hughes Institute), Fijianolide B, Laulimalide, SPA-2 (Parker Hughes Institute), SPA-1 (Parker Hughes Institute, i.e. SPIKET-P), 3-IAABU (Cytoskeleton / Mt. Sinai School of Medicine, i.e. MF-569), Narcosine (alsoPATENT Attorney Docket No.048537-668001WO known as NSC-5366), Nascapine, D-24851 (Asta Medica), A-105972 (Abbott), Hemiasterlin, 3- BAABU (Cytoskeleton / Mt. Sinai School of Medicine, i.e. MF-191), TMPN (Arizona State University), Vanadocene acetylacetonate, T-138026 (Tularik), Monsatrol, lnanocine (i.e. NSC- 698666), 3-IAABE (Cytoskeleton / Mt. Sinai School of Medicine), A-204197 (Abbott), T-607 (Tuiarik, i.e. T-900607), RPR-115781 (Aventis), Eleutherobins (such as Desmethyleleutherobin, Desaetyleleutherobin, lsoeleutherobin A, and Z-Eleutherobin), Caribaeoside, Caribaeolin, Halichondrin B, D-64131 (Asta Medica), D-68144 (Asta Medica), Diazonamide A, A-293620 (Abbott), NPI-2350 (Nereus), Taccalonolide A, TUB-245 (Aventis), A-259754 (Abbott), Diozostatin, (-)-Phenylahistin (i.e. NSCL-96F037), D-68838 (Asta Medica), D-68836 (Asta Medica), Myoseverin B, D-43411 (Zentaris, i.e. D-81862), A-289099 (Abbott), A-318315 (Abbott), HTI-286 (i.e. SPA-110, trifluoroacetate salt) (Wyeth), D-82317 (Zentaris), D-82318 (Zentaris), SC- 12983 (NCI), Resverastatin phosphate sodium, BPR-OY-007 (National Health Research Institutes), and SSR-250411 (Sanofi)), steroids (e.g., dexamethasone), finasteride, aromatase inhibitors, gonadotropin-releasing hormone agonists (GnRH) such as goserelin or leuprolide, adrenocorticosteroids (e.g., prednisone), progestins (e.g., hydroxyprogesterone caproate, megestrol acetate, medroxyprogesterone acetate), estrogens (e.g., diethlystilbestrol, ethinyl estradiol), antiestrogen (e.g., tamoxifen), androgens (e.g., testosterone propionate, fluoxymesterone), antiandrogen (e.g., flutamide), immunostimulants (e.g., Bacillus Calmette-Guérin (BCG), levamisole, interleukin-2, alpha-interferon, etc.), monoclonal antibodies (e.g., anti-CD20, anti- HER2, anti-CD52, anti-HLA-DR, and anti-VEGF monoclonal antibodies), immunotoxins (e.g., anti- CD33 monoclonal antibody-calicheamicin conjugate, anti-CD22 monoclonal antibody-pseudomonas exotoxin conjugate, etc.), radioimmunotherapy (e.g., anti-CD20 monoclonal antibody conjugated to111In,90Y, or131I, etc.), triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, topotecan, itraconazole, vindesine, cerivastatin, vincristine, deoxyadenosine, sertraline, pitavastatin, irinotecan, clofazimine, 5-nonyloxytryptamine, vemurafenib, dabrafenib, erlotinib, gefitinib, EGFR inhibitors, epidermal growth factor receptor (EGFR)-targeted therapy or therapeutic (e.g. gefitinib (Iressa ™), erlotinib (Tarceva ™), cetuximab (Erbitux™), lapatinib (Tykerb™), panitumumab (Vectibix™), vandetanib (Caprelsa™), afatinib / BIBW2992, CI-1033 / canertinib, neratinib / HKI-272, CP-724714, TAK-285, AST-1306, ARRY334543, ARRY-380, AG-1478, dacomitinib / PF299804,PATENT Attorney Docket No.048537-668001WO OSI-420 / desmethyl erlotinib, AZD8931, AEE788, pelitinib / EKB-569, CUDC-101, WZ8040, WZ4002, WZ3146, AG-490, XL647, PD153035, BMS-599626), sorafenib, imatinib, sunitinib, dasatinib, or the like.

[0089] “Selective” or “selectivity” or the like of a compound refers to the compound’s ability to discriminate between molecular targets (e.g. a compound having selectivity toward ROR1).

[0090] “Specific”, “specifically”, “specificity”, or the like of a compound refers to the compound’s ability to cause a particular action, such as inhibition, to a particular molecular target with minimal or no action to other proteins in the cell.

[0091] It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes. METHODS Methods of Manufacturing CAR-T cells

[0092] Provided herein are methods for making mycophenolate-resistant CAR-T cells. In embodiments, the T cells may be isolated or derived from various sources including, cord blood. The inventors unexpectedly discovered that a genetic engineering approach including CAR transfection, introduction of a mutated inosine monophosphate dehydrogenase 2 (IMPDH2) gene conferring mycophenolate resistance, and T-cell receptor (TCR) gene inactivation enabled robust expansion of CAR-T cells. In embodiments, the robust expansion of CAR-T cells occurred even in the presence of mycophenolate. In embodiments, the methods provided herein including embodiments thereof yields a highly enriched population of functional mycophenolate resistant CAR-T cells with reduced alloreactivity and enhanced therapeutic potential, particularly in settings requiring immunosuppression.

[0093] Thus, in an aspect is provided a method of producing an expanded population of mycophenolate resistant CAR-T cells, the method including: (i) contacting a plurality of T cells withPATENT Attorney Docket No.048537-668001WO a first expression vector including a first nucleic acid sequence encoding a chimeric antigen receptor (CAR) thereby producing a plurality of CAR-T cells and expanding said plurality of CAR-T cells for a first period of time thereby producing an expanded population of CAR-T cells, wherein the first period of time is at least 14 days; (ii) contacting said expanded population of CAR-T cells with a second expression vector including a second nucleic acid encoding a mutated IMPDH2 protein thereby producing a plurality of IMPDH2 mutant CAR-T cells, wherein said mutated IMPDH2 protein confers mycophenolate resistance to said plurality of IMPDH2 mutant CAR-T cells and expanding said plurality of IMPDH2 mutant CAR-T cells in the presence of mycophenolate for a second period of time thereby producing an expanded population of IMPDH2 CAR-T cells, wherein the second period of time is at least 14 days; and (iii) genetically altering said expanded population of IMPDH2 mutant CAR-T cells to inactivate a T-cell receptor (TCR) gene within said IMPDH2 mutant CAR-T cells thereby producing a plurality of TCR knockout IMPDH2 mutant CAR-T cells and expanding said plurality of TCR knockout IMPDH2 mutant CAR-T cells for a third period of time thereby producing an expanded population of mycophenolate resistant CAR-T cells, wherein the third period of time is at least 2 days.

[0094] In embodiments, the expanding of the plurality of TCR knockout IMPDH2 mutant CAR-T cells in step (iii) occurs in the presence of mycophenolate.

[0095] In embodiments, the step (i), step (ii), and step (iii) may occur in any order.

[0096] In another aspect is provided a method of producing an expanded population of mycophenolate resistant CAR-T cells, the method including: (i) contacting a plurality of T cells with a first expression vector including a first nucleic acid sequence encoding a mutated IMPDH2 protein thereby producing a plurality of IMPDH2 mutant T cells, wherein said mutated IMPDH2 protein confers mycophenolate resistance to said plurality of IMPDH2 mutant T cells and expanding said plurality of IMPDH2 mutant T cells in the presence of mycophenolate for a first period of time thereby producing an expanded population of IMPDH2 T cells, wherein the first period of time is at least 2 days; (ii) contacting said expanded population of IMPDH2 mutant T cells with a second expression vector including a second nucleic acid encoding a chimeric antigen receptor (CAR) thereby producing a plurality of IMPDH2 mutant CAR-T cells and expanding said plurality ofPATENT Attorney Docket No.048537-668001WO IMPDH2 mutant CAR-T cells for a second period of time thereby producing an expanded population of IMPDH2 CAR-T cells, wherein the second period of time is at least 2 days; and (iii) genetically altering said expanded population of IMPDH2 mutant CAR-T cells to inactivate a T-cell receptor (TCR) gene within said IMPDH2 mutant CAR-T cells thereby producing a plurality of TCR knockout IMPDH2 mutant CAR-T cells and expanding said plurality of TCR knockout IMPDH2 mutant CAR-T cells for a third period of time thereby producing an expanded population of mycophenolate resistant CAR-T cells, wherein the third period of time is at least 2 days.

[0097] In embodiments, the expanding said plurality of IMPDH2 mutant CAR-T cells of step (ii) occurs in the presence of mycophenolate. In embodiments, the expanding said plurality of TCR knockout IMPDH2 mutant CAR-T cells of step (iii) occurs in the presence of mycophenolate.

[0098] In another aspect is provided a method of producing an expanded population of mycophenolate resistant CAR-T cells, the method including: (i) contacting a plurality of T cells with a first expression vector including a first nucleic acid sequence encoding a mutated IMPDH2 protein thereby producing a plurality of IMPDH2 mutant T cells, wherein said mutated IMPDH2 protein confers mycophenolate resistance to said plurality of IMPDH2 mutant T cells and expanding said plurality of IMPDH2 mutant T cells in the presence of mycophenolate for a first period of time thereby producing an expanded population of IMPDH2 T cells, wherein the first period of time is at least 2 days; (ii) genetically altering said expanded population of IMPDH2 mutant T cells to inactivate a T-cell receptor (TCR) gene within said IMPDH2 mutant T cells thereby producing a plurality of TCR knockout IMPDH2 mutant T cells and expanding said plurality of TCR knockout IMPDH2 mutant T cells for a second period of time thereby producing an expanded population of TCR knockout IMPDH2 mutant T cells, wherein the second period of time is at least 2 days; and (iii) contacting said expanded population of TCR knockout IMPDH2 mutant T cells with a second expression vector including a second nucleic acid encoding a chimeric antigen receptor (CAR) thereby producing a plurality of TCR knockout IMPDH2 mutant CAR-T cells and expanding said plurality of TCR knockout IMPDH2 mutant CAR-T cells for a third period of time thereby producing an expanded population of mycophenolate resistant CAR-T cells, wherein the third period of time is at least 2 days.PATENT Attorney Docket No.048537-668001WO

[0099] In embodiments, the expanding said plurality of TCR knockout IMPDH2 mutant T cells of step (ii) occurs in the presence of mycophenolate. In embodiments, the expanding said plurality of TCR knockout IMPDH2 mutant CAR-T cells of step (iii) occurs in the presence of mycophenolate.

[0100] In another aspect is provided a method of producing an expanded population of mycophenolate resistant CAR-T cells, the method including: (i) genetically altering a plurality of T cells to inactivate a T-cell receptor (TCR) gene within said plurality of T cells thereby producing a plurality of TCR knockout T cells and expanding said plurality of TCR knockout T cells for a first period of time thereby producing an expanded population of TCR knockout T cells, wherein the first period of time is at least 2 days; (ii) contacting said expanded population of TCR knockout T cells with a first expression vector including a first nucleic acid encoding a mutated IMPDH2 protein thereby producing a plurality of TCR knockout IMPDH2 mutant T cells, wherein said mutated IMPDH2 protein confers mycophenolate resistance to said plurality of TCR knockout IMPDH2 mutant T cells and expanding said plurality of TCR knockout IMPDH2 mutant T cells in the presence of mycophenolate for a second period of time thereby producing an expanded population of TCR knockout IMPDH2 mutant T cells, wherein the second period of time is at least 2 days; and (iii) contacting said expanded population of TCR knockout IMPDH2 mutant T cells with a second expression vector including a second nucleic acid sequence encoding a chimeric antigen receptor (CAR) thereby producing a plurality of TCR knockout IMPDH2 mutant CAR-T cells and expanding said plurality of TCR knockout IMPDH2 mutant CAR-T cells for a third period of time thereby producing an expanded population of mycophenolate resistant CAR-T cells, wherein the first period of time is at least 2 days.

[0101] In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 35 µg / ml. In embodiments, thePATENT Attorney Docket No.048537-668001WO mycophenolate is present in an amount between about 100 ng / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 1 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 1 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 2 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 3 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 4 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 5 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 6 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 7 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 8 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 9 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 10 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 11 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 12 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 13 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 14 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 15 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 16 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 17 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 18 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 19 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 20 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 21 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 22 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is presentPATENT Attorney Docket No.048537-668001WO in an amount between about 23 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 24 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 25 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 26 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 27 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 28 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 29 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 30 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 31 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 32 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 33 µg / ml and about 35 µg / ml. In embodiments, the mycophenolate is present in an amount between about 34 µg / ml and about 35 µg / ml.

[0102] In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 34 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 33 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 32 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 31 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 30 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 29 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 28 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 27 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 26 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 25 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 24 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 23 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 22 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 21 µg / ml. In embodiments, thePATENT Attorney Docket No.048537-668001WO mycophenolate is present in an amount between about 100 ng / ml and about 20 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 19 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 18 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 17 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 16 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 15 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 14 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 13 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 12 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 11 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 10 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 9 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 8 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 7 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 6 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 5 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 4 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 3 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 2 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 1 µg / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 900 ng / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 800 ng / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 700 ng / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 600 ng / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 500 ng / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 400 ng / ml. InPATENT Attorney Docket No.048537-668001WO embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 300 ng / ml. In embodiments, the mycophenolate is present in an amount between about 100 ng / ml and about 200 ng / ml.

[0103] In embodiments, the mycophenolate is present in an amount of about 100 ng / ml. In embodiments, the mycophenolate is present in an amount of about 200 ng / ml. In embodiments, the mycophenolate is present in an amount of about 300 ng / ml. In embodiments, the mycophenolate is present in an amount of about 400 ng / ml. In embodiments, the mycophenolate is present in an amount of about 500 ng / ml. In embodiments, the mycophenolate is present in an amount of about 600 ng / ml. In embodiments, the mycophenolate is present in an amount of about 700 ng / ml. In embodiments, the mycophenolate is present in an amount of about 800 ng / ml. In embodiments, the mycophenolate is present in an amount of about 900 ng / ml. In embodiments, the mycophenolate is present in an amount of about 1 µg / ml. In embodiments, the mycophenolate is present in an amount of about 2 µg / ml. In embodiments, the mycophenolate is present in an amount of about 3 µg / ml. In embodiments, the mycophenolate is present in an amount of about 4 µg / ml. In embodiments, the mycophenolate is present in an amount of about 5 µg / ml. In embodiments, the mycophenolate is present in an amount of about 6 µg / ml. In embodiments, the mycophenolate is present in an amount of about 7 µg / ml. In embodiments, the mycophenolate is present in an amount of about 8 µg / ml. In embodiments, the mycophenolate is present in an amount of about 9 µg / ml. In embodiments, the mycophenolate is present in an amount of about 10 µg / ml. In embodiments, the mycophenolate is present in an amount of about 11 µg / ml. In embodiments, the mycophenolate is present in an amount of about 12 µg / ml. In embodiments, the mycophenolate is present in an amount of about 13 µg / ml. In embodiments, the mycophenolate is present in an amount of about 14 µg / ml. In embodiments, the mycophenolate is present in an amount of about 15 µg / ml. In embodiments, the mycophenolate is present in an amount of about 16 µg / ml. In embodiments, the mycophenolate is present in an amount of about 17 µg / ml. In embodiments, the mycophenolate is present in an amount of about 18 µg / ml. In embodiments, the mycophenolate is present in an amount of about 19 µg / ml. In embodiments, the mycophenolate is present in an amount of about 20 µg / ml. In embodiments, the mycophenolate is present in an amount of about 21 µg / ml. In embodiments, the mycophenolate is present in an amount of about 22 µg / ml. In embodiments, the mycophenolate isPATENT Attorney Docket No.048537-668001WO present in an amount of about 23 µg / ml. In embodiments, the mycophenolate is present in an amount of about 24 µg / ml. In embodiments, the mycophenolate is present in an amount of about 25 µg / ml. In embodiments, the mycophenolate is present in an amount of about 26 µg / ml. In embodiments, the mycophenolate is present in an amount of about 27 µg / ml. In embodiments, the mycophenolate is present in an amount of about 28 µg / ml. In embodiments, the mycophenolate is present in an amount of about 29 µg / ml. In embodiments, the mycophenolate is present in an amount of about 30 µg / ml. In embodiments, the mycophenolate is present in an amount of about 31 µg / ml. In embodiments, the mycophenolate is present in an amount of about 32 µg / ml. In embodiments, the mycophenolate is present in an amount of about 33 µg / ml. In embodiments, the mycophenolate is present in an amount of about 34 µg / ml. In embodiments, the mycophenolate is present in an amount of about 35 µg / ml.

[0104] In embodiments, the plurality of T cells is isolated from cord blood prior to step (i). In embodiments, the plurality of T cells is derived from cord blood prior to step (i). In embodiments, the plurality of T cells is isolated from peripheral blood prior to step (i). In embodiments, the plurality of T cells is isolated from peripheral blood from neonates, children, or adults prior to step (i). In embodiments, the plurality of T cells is isolated from peripheral blood from neonates prior to step (i). In embodiments, the plurality of T cells is isolated from peripheral blood from children prior to step (i). In embodiments, the plurality of T cells is isolated from peripheral blood from adults prior to step (i). In embodiments, the plurality of T cells is derived from peripheral blood prior to step (i). In embodiments, the plurality of T cells is derived from peripheral blood from neonates, children, or adults prior to step (i).

[0105] In embodiments, the expanded population of mycophenolate resistant CAR-T cells is at least 10 times more cells than the expanded population of IMPDH2 mutant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells is at least 15 times more cells than the expanded population of IMPDH2 mutant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells is at least 20 times more cells than the expanded population of IMPDH2 mutant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells is at least 25 times more cells than the expanded population ofPATENT Attorney Docket No.048537-668001WO IMPDH2 mutant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells is at least 30 times more cells than the expanded population of IMPDH2 mutant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells is at least 35 times more cells than the expanded population of IMPDH2 mutant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells is at least 40 times more cells than the expanded population of IMPDH2 mutant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells is at least 45 times more cells than the expanded population of IMPDH2 mutant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells is at least 50 times more cells than the expanded population of IMPDH2 mutant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells is at least 55 times more cells than the expanded population of IMPDH2 mutant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells is at least 60 times more cells than the expanded population of IMPDH2 mutant CAR-T cells.

[0106] In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1.5 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 2 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 2.5 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 3 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 3.5 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 4 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 4.5 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population ofPATENT Attorney Docket No.048537-668001WO mycophenolate resistant CAR-T cells includes between about 5 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 5.5 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 6 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 6.5 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 7 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 7.5 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 8 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 8.5 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 9 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 9.5 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells.

[0107] In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1.5 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 2 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 2.5 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 3 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 3.5 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. InPATENT Attorney Docket No.048537-668001WO embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 4 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 4.5 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 5 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 5.5 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 6 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 6.5 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 7 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 7.5 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 8 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 8.5 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 9 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 9.5 x 1013and about 8 x 1014mycophenolate resistant CAR-T cells.

[0108] In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1.5 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 2 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 2.5 x 1014and about 8 x 1014PATENT Attorney Docket No.048537-668001WO mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 3 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 3.5 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 4 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 4.5 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 5 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 5.5 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 6 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 6.5 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 7 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 7.5 x 1014and about 8 x 1014mycophenolate resistant CAR-T cells.

[0109] In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 7.5 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 7 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 6.5 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 6 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cellsPATENT Attorney Docket No.048537-668001WO includes between about 1 x 1012and about 5.5 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 5 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 4.5 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 4 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 3.5 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 3 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 2.5 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 2 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 1.5 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 1 x 1014mycophenolate resistant CAR-T cells.

[0110] In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 9.5 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 9 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 8.5 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 8 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 7.5 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 7 x 1013mycophenolate resistant CAR-T cells. InPATENT Attorney Docket No.048537-668001WO embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 6.5 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 6 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 5.5 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 5 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 4.5 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 4 x 1014mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 3.5 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 3 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 2.5 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 2 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 1.5 x 1013mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 1 x 1013mycophenolate resistant CAR-T cells.

[0111] In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 9.5 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 9 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 8.5 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 8 x 1012PATENT Attorney Docket No.048537-668001WO mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 7.5 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 7 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 6.5 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 6 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 5.5 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 5 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 4.5 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 4 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 3.5 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 3 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 2.5 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 2 x 1012mycophenolate resistant CAR-T cells. In embodiments, the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 1012and about 1.5 x 1013mycophenolate resistant CAR-T cells.

[0112] In embodiments, the first period of time is at least 3 days. In embodiments, the first period of time is at least 4 days. In embodiments, the first period of time is at least 5 days. In embodiments, the first period of time is at least 6 days. In embodiments, the first period of time is at least 7 days. In embodiments, the first period of time is at least 8 days. In embodiments, the first period of time is at least 9 days. In embodiments, the first period of time is at least 10 days. In embodiments, the firstPATENT Attorney Docket No.048537-668001WO period of time is at least 11 days. In embodiments, the first period of time is at least 12 days. In embodiments, the first period of time is at least 13 days. In embodiments, the first period of time is at least 14 days. In embodiments, the first period of time is at least 15 days. In embodiments, the first period of time is at least 16 days. In embodiments, the first period of time is at least 17 days. In embodiments, the first period of time is at least 18 days. In embodiments, the first period of time is at least 19 days. In embodiments, the first period of time is at least 20 days. In embodiments, the first period of time is at least 21 days.

[0113] In embodiments, the first period of time is for at least 4 weeks. In embodiments, the first period of time is for at least 5 weeks. In embodiments, the first period of time is for at least 6 weeks. In embodiments, the first period of time is for at least 7 weeks. In embodiments, the first period of time is for at least 8 weeks. In embodiments, the first period of time is for at least 9 weeks. In embodiments, the first period of time is for at least 10 weeks.

[0114] In embodiments, the first period of time is between about 4 weeks and about 10 weeks. In embodiments, the first period of time is between about 5 weeks and about 10 weeks. In embodiments, the first period of time is between about 6 weeks and about 10 weeks. In embodiments, the first period of time is between about 7 weeks and about 10 weeks. In embodiments, the first period of time is between about 8 weeks and about 10 weeks. In embodiments, the first period of time is between about 9 weeks and about 10 weeks.

[0115] In embodiments, the first period of time is between about 4 weeks and about 9 weeks. In embodiments, the first period of time is between about 4 weeks and about 8 weeks. In embodiments, the first period of time is between about 4 weeks and about 7 weeks. In embodiments, the first period of time is between about 4 weeks and about 6 weeks. In embodiments, the first period of time is between about 4 weeks and about 5 weeks.

[0116] In embodiments, the first period of time is between 4 weeks and 10 weeks. In embodiments, the first period of time is between 5 weeks and 10 weeks. In embodiments, the first period of time is between 6 weeks and 10 weeks. In embodiments, the first period of time is betweenPATENT Attorney Docket No.048537-668001WO 7 weeks and 10 weeks. In embodiments, the first period of time is between 8 weeks and 10 weeks. In embodiments, the first period of time is between 9 weeks and 10 weeks.

[0117] In embodiments, the first period of time is between 4 weeks and 9 weeks. In embodiments, the first period of time is between 4 weeks and 8 weeks. In embodiments, the first period of time is between 4 weeks and 7 weeks. In embodiments, the first period of time is between 4 weeks and 6 weeks. In embodiments, the first period of time is between 4 weeks and 5 weeks.

[0118] In embodiments, the first period of time is about 4 weeks. In embodiments, the first period of time is about 5 weeks. In embodiments, the first period of time about 6 weeks. In embodiments, the first period of time is about 7 weeks. In embodiments, the first period of time is about 8 weeks. In embodiments, the first period of time is about 9 weeks. In embodiments, the first period of time is about 10 weeks.

[0119] In embodiments, the first period of time is 4 weeks. In embodiments, the first period of time is 5 weeks. In embodiments, the first period of time 6 weeks. In embodiments, the first period of time is 7 weeks. In embodiments, the first period of time is 8 weeks. In embodiments, the first period of time is 9 weeks. In embodiments, the first period of time is 10 weeks.

[0120] In embodiments, the second period of time is at least 3 days. In embodiments, the second period of time is at least 4 days. In embodiments, the second period of time is at least 5 days. In embodiments, the second period of time is at least 6 days. In embodiments, the second period of time is at least 7 days. In embodiments, the second period of time is at least 8 days. In embodiments, the second period of time is at least 9 days. In embodiments, the second period of time is at least 10 days. In embodiments, the second period of time is at least 11 days. In embodiments, the second period of time is at least 12 days. In embodiments, the second period of time is at least 13 days. In embodiments, the second period of time is at least 14 days. In embodiments, the second period of time is at least 15 days. In embodiments, the second period of time is at least 16 days. In embodiments, the second period of time is at least 17 days. In embodiments, the second period of time is at least 18 days. In embodiments, the second period of time is at least 19 days. InPATENT Attorney Docket No.048537-668001WO embodiments, the second period of time is at least 20 days. In embodiments, the second period of time is at least 21 days.

[0121] In embodiments, the second period of time is for at least 4 weeks. In embodiments, the second period of time is for at least 5 weeks. In embodiments, the second period of time is for at least 6 weeks. In embodiments, the second period of time is for at least 7 weeks. In embodiments, the second period of time is for at least 8 weeks. In embodiments, the second period of time is for at least 9 weeks. In embodiments, the second period of time is for at least 10 weeks.

[0122] In embodiments, the second period of time is between about 4 weeks and about 10 weeks. In embodiments, the second period of time is between about 5 weeks and about 10 weeks. In embodiments, the second period of time is between about 6 weeks and about 10 weeks. In embodiments, the second period of time is between about 7 weeks and about 10 weeks. In embodiments, the second period of time is between about 8 weeks and about 10 weeks. In embodiments, the second period of time is between about 9 weeks and about 10 weeks.

[0123] In embodiments, the second period of time is between about 4 weeks and about 9 weeks. In embodiments, the second period of time is between about 4 weeks and about 8 weeks. In embodiments, the second period of time is between about 4 weeks and about 7 weeks. In embodiments, the second period of time is between about 4 weeks and about 6 weeks. In embodiments, the second period of time is between about 4 weeks and about 5 weeks.

[0124] In embodiments, the second period of time is between 4 weeks and 10 weeks. In embodiments, the second period of time is between 5 weeks and 10 weeks. In embodiments, the second period of time is between 6 weeks and 10 weeks. In embodiments, the second period of time is between 7 weeks and 10 weeks. In embodiments, the second period of time is between 8 weeks and 10 weeks. In embodiments, the second period of time is between 9 weeks and 10 weeks.

[0125] In embodiments, the second period of time is between 4 weeks and 9 weeks. In embodiments, the second period of time is between 4 weeks and 8 weeks. In embodiments, the second period of time is between 4 weeks and 7 weeks. In embodiments, the second period of timePATENT Attorney Docket No.048537-668001WO is between 4 weeks and 6 weeks. In embodiments, the second period of time is between 4 weeks and 5 weeks.

[0126] In embodiments, the second period of time is about 4 weeks. In embodiments, the second period of time is about 5 weeks. In embodiments, the second period of time about 6 weeks. In embodiments, the second period of time is about 7 weeks. In embodiments, the second period of time is about 8 weeks. In embodiments, the second period of time is about 9 weeks. In embodiments, the second period of time is about 10 weeks.

[0127] In embodiments, the second period of time is 4 weeks. In embodiments, the second period of time is 5 weeks. In embodiments, the second period of time 6 weeks. In embodiments, the second period of time is 7 weeks. In embodiments, the second period of time is 8 weeks. In embodiments, the second period of time is 9 weeks. In embodiments, the second period of time is 10 weeks.

[0128] In embodiments, the third period of time is between about 2 days and about 60 days. In embodiments, the third period of time is between about 3 days and about 60 days. In embodiments, the third period of time is between about 4 days and about 60 days. In embodiments, the third period of time is between about 5 days and about 60 days. In embodiments, the third period of time is between about 6 days and about 60 days. In embodiments, the third period of time is between about 7 days and about 60 days. In embodiments, the third period of time is between about 8 days and about 60 days. In embodiments, the third period of time is between about 9 days and about 60 days. In embodiments, the third period of time is between about 10 days and about 60 days. In embodiments, the third period of time is between about 11 days and about 60 days. In embodiments, the third period of time is between about 12 days and about 60 days. In embodiments, the third period of time is between about 13 days and about 60 days. In embodiments, the third period of time is between about 14 days and about 60 days. In embodiments, the third period of time is between about 15 days and about 60 days. In embodiments, the third period of time is between about 16 days and about 60 days. In embodiments, the third period of time is between about 17 days and about 60 days. In embodiments, the third period of time is between about 18 days and about 60 days. In embodiments, the third period of time is between about 19 days and about 60 days. In embodiments, the third period of time is between about 20 days and about 60 days. In embodiments, the thirdPATENT Attorney Docket No.048537-668001WO period of time is between about 21 days and about 60 days. In embodiments, the third period of time is between about 22 days and about 60 days. In embodiments, the third period of time is between about 23 days and about 60 days. In embodiments, the third period of time is between about 24 days and about 60 days. In embodiments, the third period of time is between about 25 days and about 60 days. In embodiments, the third period of time is between about 26 days and about 60 days. In embodiments, the third period of time is between about 27 days and about 60 days. In embodiments, the third period of time is between about 28 days and about 60 days. In embodiments, the third period of time is between about 29 days and about 60 days.

[0129] In embodiments, the third period of time is between about 30 days and about 60 days. In embodiments, the third period of time is between about 31 days and about 60 days. In embodiments, the third period of time is between about 32 days and about 60 days. In embodiments, the third period of time is between about 33 days and about 60 days. In embodiments, the third period of time is between about 34 days and about 60 days. In embodiments, the third period of time is between about 35 days and about 60 days. In embodiments, the third period of time is between about 36 days and about 60 days. In embodiments, the third period of time is between about 37 days and about 60 days. In embodiments, the third period of time is between about 38 days and about 60 days. In embodiments, the third period of time is between about 39 days and about 60 days. In embodiments, the third period of time is between about 40 days and about 60 days. In embodiments, the third period of time is between about 41 days and about 60 days. In embodiments, the third period of time is between about 42 days and about 60 days. In embodiments, the third period of time is between about 43 days and about 60 days. In embodiments, the third period of time is between about 44 days and about 60 days. In embodiments, the third period of time is between about 45 days and about 60 days. In embodiments, the third period of time is between about 46 days and about 60 days. In embodiments, the third period of time is between about 47 days and about 60 days. In embodiments, the third period of time is between about 48 days and about 60 days. In embodiments, the third period of time is between about 49 days and about 60 days. In embodiments, the third period of time is between about 50 days and about 60 days. In embodiments, the third period of time is between about 51 days and about 60 days. In embodiments, the third period of time is between about 52 days and about 60 days. In embodiments, the third period of time is between about 53 days and about 60PATENT Attorney Docket No.048537-668001WO days. In embodiments, the third period of time is between about 54 days and about 60 days. In embodiments, the third period of time is between about 55 days and about 60 days. In embodiments, the third period of time is between about 56 days and about 60 days. In embodiments, the third period of time is between about 57 days and about 60 days. In embodiments, the third period of time is between about 58 days and about 60 days. In embodiments, the third period of time is between about 59 days and about 60 days.

[0130] In embodiments, the third period of time is between about 2 days and about 59 days. In embodiments, the third period of time is between about 2 days and about 58 days. In embodiments, the third period of time is between about 2 days and about 57 days. In embodiments, the third period of time is between about 2 days and about 56 days. In embodiments, the third period of time is between about 2 days and about 55 days. In embodiments, the third period of time is between about 2 days and about 54 days. In embodiments, the third period of time is between about 2 days and about 53 days. In embodiments, the third period of time is between about 2 days and about 52 days. In embodiments, the third period of time is between about 2 days and about 51 days. In embodiments, the third period of time is between about 2 days and about 50 days. In embodiments, the third period of time is between about 2 days and about 49 days. In embodiments, the third period of time is between about 2 days and about 48 days. In embodiments, the third period of time is between about 2 days and about 47 days. In embodiments, the third period of time is between about 2 days and about 46 days. In embodiments, the third period of time is between about 2 days and about 45 days. In embodiments, the third period of time is between about 2 days and about 44 days. In embodiments, the third period of time is between about 2 days and about 43 days. In embodiments, the third period of time is between about 2 days and about 42 days. In embodiments, the third period of time is between about 2 days and about 41 days. In embodiments, the third period of time is between about 2 days and about 40 days. In embodiments, the third period of time is between about 2 days and about 39 days. In embodiments, the third period of time is between about 2 days and about 38 days. In embodiments, the third period of time is between about 2 days and about 37 days. In embodiments, the third period of time is between about 2 days and about 36 days. In embodiments, the third period of time is between about 2 days and about 35 days. In embodiments, the third period of time is between about 2 days and about 34 days. In embodiments,PATENT Attorney Docket No.048537-668001WO the third period of time is between about 2 days and about 33 days. In embodiments, the third period of time is between about 2 days and about 32 days. In embodiments, the third period of time is between about 2 days and about 31 days. In embodiments, the third period of time is between about 2 days and about 30 days.

[0131] In embodiments, the third period of time is between about 2 days and about 29 days. In embodiments, the third period of time is between about 2 days and about 28 days. In embodiments, the third period of time is between about 2 days and about 27 days. In embodiments, the third period of time is between about 2 days and about 26 days. In embodiments, the third period of time is between about 2 days and about 25 days. In embodiments, the third period of time is between about 2 days and about 24 days. In embodiments, the third period of time is between about 2 days and about 23 days. In embodiments, the third period of time is between about 2 days and about 22 days. In embodiments, the third period of time is between about 2 days and about 21 days. In embodiments, the third period of time is between about 2 days and about 20 days. In embodiments, the third period of time is between about 2 days and about 19 days. In embodiments, the third period of time is between about 2 days and about 18 days. In embodiments, the third period of time is between about 2 days and about 17 days. In embodiments, the third period of time is between about 2 days and about 16 days. In embodiments, the third period of time is between about 2 days and about 15 days. In embodiments, the third period of time is between about 2 days and about 14 days. In embodiments, the third period of time is between about 2 days and about 13 days. In embodiments, the third period of time is between about 2 days and about 12 days. In embodiments, the third period of time is between about 2 days and about 11 days. In embodiments, the third period of time is between about 2 days and about 10 days. In embodiments, the third period of time is between about 2 days and about 9 days. In embodiments, the third period of time is between about 2 days and about 8 days. In embodiments, the third period of time is between about 2 days and about 7 days. In embodiments, the third period of time is between about 2 days and about 6 days. In embodiments, the third period of time is between about 2 days and about 5 days. In embodiments, the third period of time is between about 2 days and about 4 days. In embodiments, the third period of time is between about 2 days and about 3 days.PATENT Attorney Docket No.048537-668001WO

[0132] In embodiments, the third period of time is between 2 days and 60 days. In embodiments, the third period of time is between 3 days and 60 days. In embodiments, the third period of time is between 4 days and 60 days. In embodiments, the third period of time is between 5 days and 60 days. In embodiments, the third period of time is between 6 days and 60 days. In embodiments, the third period of time is between 7 days and 60 days. In embodiments, the third period of time is between 8 days and 60 days. In embodiments, the third period of time is between 9 days and 60 days. In embodiments, the third period of time is between 10 days and 60 days. In embodiments, the third period of time is between 11 days and 60 days. In embodiments, the third period of time is between 12 days and 60 days. In embodiments, the third period of time is between 13 days and 60 days. In embodiments, the third period of time is between 14 days and 60 days. In embodiments, the third period of time is between 15 days and 60 days. In embodiments, the third period of time is between 16 days and 60 days. In embodiments, the third period of time is between 17 days and 60 days. In embodiments, the third period of time is between 18 days and 60 days. In embodiments, the third period of time is between 19 days and 60 days. In embodiments, the third period of time is between 20 days and 60 days. In embodiments, the third period of time is between 21 days and 60 days. In embodiments, the third period of time is between 22 days and 60 days. In embodiments, the third period of time is between 23 days and 60 days. In embodiments, the third period of time is between 24 days and 60 days. In embodiments, the third period of time is between 25 days and 60 days. In embodiments, the third period of time is between 26 days and 60 days. In embodiments, the third period of time is between 27 days and 60 days. In embodiments, the third period of time is between 28 days and 60 days. In embodiments, the third period of time is between 29 days and 60 days.

[0133] In embodiments, the third period of time is between 30 days and 60 days. In embodiments, the third period of time is between 31 days and 60 days. In embodiments, the third period of time is between 32 days and 60 days. In embodiments, the third period of time is between 33 days and 60 days. In embodiments, the third period of time is between 34 days and 60 days. In embodiments, the third period of time is between 35 days and 60 days. In embodiments, the third period of time is between 36 days and 60 days. In embodiments, the third period of time is between 37 days and 60 days. In embodiments, the third period of time is between 38 days and 60 days. In embodiments, thePATENT Attorney Docket No.048537-668001WO third period of time is between 39 days and 60 days. In embodiments, the third period of time is between 40 days and 60 days. In embodiments, the third period of time is between 41 days and 60 days. In embodiments, the third period of time is between 42 days and 60 days. In embodiments, the third period of time is between 43 days and 60 days. In embodiments, the third period of time is between 44 days and 60 days. In embodiments, the third period of time is between 40 days and 60 days. In embodiments, the third period of time is between 45 days and 60 days. In embodiments, the third period of time is between 46 days and 60 days. In embodiments, the third period of time is between 47 days and 60 days. In embodiments, the third period of time is between 48 days and 60 days. In embodiments, the third period of time is between 49 days and 60 days. In embodiments, the third period of time is between 50 days and 60 days. In embodiments, the third period of time is between 51 days and 60 days. In embodiments, the third period of time is between 52 days and 60 days. In embodiments, the third period of time is between 53 days and 60 days. In embodiments, the third period of time is between 54 days and 60 days. In embodiments, the third period of time is between 55 days and 60 days. In embodiments, the third period of time is between 56 days and 60 days. In embodiments, the third period of time is between 57 days and 60 days. In embodiments, the third period of time is between 58 days and 60 days. In embodiments, the third period of time is between 59 days and 60 days.

[0134] In embodiments, the third period of time is between 2 days and 59 days. In embodiments, the third period of time is between 2 days and 58 days. In embodiments, the third period of time is between 2 days and 57 days. In embodiments, the third period of time is between 2 days and 56 days. In embodiments, the third period of time is between 2 days and 55 days. In embodiments, the third period of time is between 2 days and 54 days. In embodiments, the third period of time is between 2 days and 53 days. In embodiments, the third period of time is between 2 days and 52 days. In embodiments, the third period of time is between 2 days and 51 days. In embodiments, the third period of time is between 2 days and 50 days. In embodiments, the third period of time is between 2 days and 49 days. In embodiments, the third period of time is between 2 days and 48 days. In embodiments, the third period of time is between 2 days and 47 days. In embodiments, the third period of time is between 2 days and 46 days. In embodiments, the third period of time is between 2 days and 45 days. In embodiments, the third period of time is between 2 days and 44PATENT Attorney Docket No.048537-668001WO days. In embodiments, the third period of time is between 2 days and 43 days. In embodiments, the third period of time is between 2 days and 42 days. In embodiments, the third period of time is between 2 days and 41 days. In embodiments, the third period of time is between 2 days and 40 days. In embodiments, the third period of time is between 2 days and 39 days. In embodiments, the third period of time is between 2 days and 38 days. In embodiments, the third period of time is between 2 days and 37 days. In embodiments, the third period of time is between 2 days and 36 days. In embodiments, the third period of time is between 2 days and 35 days. In embodiments, the third period of time is between 2 days and 34 days. In embodiments, the third period of time is between 2 days and 33 days. In embodiments, the third period of time is between 2 days and 32 days. In embodiments, the third period of time is between 2 days and 31 days. In embodiments, the third period of time is between 2 days and 30 days.

[0135] In embodiments, the third period of time is between 2 days and 29 days. In embodiments, the third period of time is between 2 days and 28 days. In embodiments, the third period of time is between 2 days and 27 days. In embodiments, the third period of time is between 2 days and 26 days. In embodiments, the third period of time is between 2 days and 25 days. In embodiments, the third period of time is between 2 days and 24 days. In embodiments, the third period of time is between 2 days and 23 days. In embodiments, the third period of time is between 2 days and 22 days. In embodiments, the third period of time is between 2 days and 21 days. In embodiments, the third period of time is between 2 days and 20 days. In embodiments, the third period of time is between 2 days and 19 days. In embodiments, the third period of time is between 2 days and 18 days. In embodiments, the third period of time is between 2 days and 17 days. In embodiments, the third period of time is between 2 days and 16 days. In embodiments, the third period of time is between 2 days and 15 days. In embodiments, the third period of time is between 2 days and 14 days. In embodiments, the third period of time is between 2 days and 13 days. In embodiments, the third period of time is between 2 days and 12 days. In embodiments, the third period of time is between 2 days and 11 days. In embodiments, the third period of time is between 2 days and 10 days. In embodiments, the third period of time is between 2 days and 9 days. In embodiments, the third period of time is between 2 days and 8 days. In embodiments, the third period of time is between 2 days and 7 days. In embodiments, the third period of time is between 2 days and 6 days.PATENT Attorney Docket No.048537-668001WO In embodiments, the third period of time is between 2 days and 5 days. In embodiments, the third period of time is between 2 days and 4 days. In embodiments, the third period of time is between 2 days and 3 days.

[0136] In embodiments, the third period of time is about 2 days. In embodiments, the third period of time is about 3 days. In embodiments, the third period of time is about 4 days. In embodiments, the third period of time is about 5 days. In embodiments, the third period of time is about 6 days. In embodiments, the third period of time is about 7 days. In embodiments, the third period of time is about 8 days. In embodiments, the third period of time is about 9 days. In embodiments, the third period of time is about 10 days. In embodiments, the third period of time is about 11 days. In embodiments, the third period of time is about 12 days. In embodiments, the third period of time is about 13 days. In embodiments, the third period of time is about 14 days. In embodiments, the third period of time is about 15 days. In embodiments, the third period of time is about 16 days. In embodiments, the third period of time is about 17 days. In embodiments, the third period of time is about 18 days. In embodiments, the third period of time is about 19 days. In embodiments, the third period of time is about 20 days. In embodiments, the third period of time is about 21 days. In embodiments, the third period of time is about 22 days. In embodiments, the third period of time is about 23 days. In embodiments, the third period of time is about 24 days. In embodiments, the third period of time is about 25 days. In embodiments, the third period of time is about 26 days. In embodiments, the third period of time is about 27 days. In embodiments, the third period of time is about 28 days. In embodiments, the third period of time is about 29 days. In embodiments, the third period of time is about 30 days. In embodiments, the third period of time is about 31 days. In embodiments, the third period of time is about 32 days. In embodiments, the third period of time is about 33 days. In embodiments, the third period of time is about 34 days. In embodiments, the third period of time is about 35 days. In embodiments, the third period of time is about 36 days. In embodiments, the third period of time is about 37 days. In embodiments, the third period of time is about 38 days. In embodiments, the third period of time is about 39 days. In embodiments, the third period of time is about 40 days. In embodiments, the third period of time is about 41 days. In embodiments, the third period of time is about 42 days. In embodiments, the third period of time is about 43 days. In embodiments, the third period of time is about 44 days. In embodiments, the thirdPATENT Attorney Docket No.048537-668001WO period of time is about 45 days. In embodiments, the third period of time is about 46 days. In embodiments, the third period of time is about 47 days. In embodiments, the third period of time is about 48 days. In embodiments, the third period of time is about 49 days. In embodiments, the third period of time is about 50 days. In embodiments, the third period of time is about 51 days. In embodiments, the third period of time is about 52 days. In embodiments, the third period of time is about 53 days. In embodiments, the third period of time is about 54 days. In embodiments, the third period of time is about 55 days. In embodiments, the third period of time is about 56 days. In embodiments, the third period of time is about 57 days. In embodiments, the third period of time is about 58 days. In embodiments, the third period of time is about 59 days. In embodiments, the third period of time is about 58 days. In embodiments, the third period of time is about 60 days.

[0137] In embodiments, the third period of time is at least 3 days. In embodiments, the third period of time is at least 4 days. In embodiments, the third period of time is at least 5 days. In embodiments, the third period of time is at least 6 days. In embodiments, the third period of time is at least 7 days. In embodiments, the third period of time is at least 8 days. In embodiments, the third period of time is at least 9 days. In embodiments, the third period of time is at least 10 days. In embodiments, the third period of time is at least 11 days. In embodiments, the third period of time is at least 12 days. In embodiments, the third period of time is at least 13 days. In embodiments, the third period of time is at least 14 days. In embodiments, the third period of time is at least 15 days. In embodiments, the third period of time is at least 16 days. In embodiments, the third period of time is at least 17 days. In embodiments, the third period of time is at least 18 days. In embodiments, the third period of time is at least 19 days. In embodiments, the third period of time is at least 20 days. In embodiments, the third period of time is at least 21 days.

[0138] In embodiments, the third period of time is 2 days. In embodiments, the third period of time is 3 days. In embodiments, the third period of time is 4 days. In embodiments, the third period of time is 5 days. In embodiments, the third period of time is 6 days. In embodiments, the third period of time is 7 days. In embodiments, the third period of time is 8 days. In embodiments, the third period of time is 9 days. In embodiments, the third period of time is 10 days. In embodiments, the third period of time is 11 days. In embodiments, the third period of time is 12 days. InPATENT Attorney Docket No.048537-668001WO embodiments, the third period of time is 13 days. In embodiments, the third period of time is 14 days. In embodiments, the third period of time is 15 days. In embodiments, the third period of time is 16 days. In embodiments, the third period of time is 17 days. In embodiments, the third period of time is 18 days. In embodiments, the third period of time is 19 days. In embodiments, the third period of time is 20 days. In embodiments, the third period of time is 21 days. In embodiments, the third period of time is 22 days. In embodiments, the third period of time is 23 days. In embodiments, the third period of time is 24 days. In embodiments, the third period of time is 25 days. In embodiments, the third period of time is 26 days. In embodiments, the third period of time is 27 days. In embodiments, the third period of time is 28 days. In embodiments, the third period of time is 29 days. In embodiments, the third period of time is 30 days. In embodiments, the third period of time is 31 days. In embodiments, the third period of time is 32 days. In embodiments, the third period of time is 33 days. In embodiments, the third period of time is 34 days. In embodiments, the third period of time is 35 days. In embodiments, the third period of time is 36 days. In embodiments, the third period of time is 37 days. In embodiments, the third period of time is about 38 days. In embodiments, the third period of time is 39 days. In embodiments, the third period of time is 40 days. In embodiments, the third period of time is 41 days. In embodiments, the third period of time is 42 days. In embodiments, the third period of time is 43 days. In embodiments, the third period of time is 44 days. In embodiments, the third period of time is 45 days. In embodiments, the third period of time is 46 days. In embodiments, the third period of time is 47 days. In embodiments, the third period of time is 48 days. In embodiments, the third period of time is 49 days. In embodiments, the third period of time is 50 days. In embodiments, the third period of time is 51 days. In embodiments, the third period of time is 52 days. In embodiments, the third period of time is 53 days. In embodiments, the third period of time is 54 days. In embodiments, the third period of time is 55 days. In embodiments, the third period of time is 56 days. In embodiments, the third period of time is 57 days. In embodiments, the third period of time is 58 days. In embodiments, the third period of time is 59 days. In embodiments, the third period of time is 58 days. In embodiments, the third period of time is 60 days.

[0139] In embodiments, the mutated IMPDH2 protein includes a catalytic site mutation. In embodiments, the mutated IMPDH2 protein includes an isoleucine substitution at a positionPATENT Attorney Docket No.048537-668001WO corresponding to position 333 of SEQ ID NO:1 and / or a tyrosine substitution at a position corresponding to position 351 of SEQ ID NO:1. In embodiments, the mutated IMPDH2 protein includes an isoleucine substitution at a position corresponding to position 333 of SEQ ID NO:1 and a tyrosine substitution at a position corresponding to position 351 of SEQ ID NO:1. In embodiments, the mutated IMPDH2 protein includes an isoleucine substitution at a position corresponding to position 333 of SEQ ID NO:1 or a tyrosine substitution at a position corresponding to position 351 of SEQ ID NO:1. In embodiments, the mutated IMPDH2 protein includes an isoleucine substitution at a position corresponding to position 333 of SEQ ID NO:1. In embodiments, the mutated IMPDH2 protein includes a tyrosine substitution at a position corresponding to position 351 of SEQ ID NO:1.

[0140] In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having atleast 85% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 86% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 87% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 88% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 89% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 91% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 92% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 93% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 94% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includesPATENT Attorney Docket No.048537-668001WO an amino acid sequence having at least 96% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 97% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 98% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein includes an amino acid sequence having at least 100% sequence identity to the amino acid sequence of SEQ ID NO:2.

[0141] In embodiments, the mutated IMPDH2 protein includes the amino acid sequence of SEQ ID NO:2. In embodiments, the mutated IMPDH2 protein has the amino acid sequence of SEQ ID NO:2.

[0142] In embodiments, the first expression vector is a CRISPR / Cas9 expression vector, a TALEN expression vector, or a viral expression vector. In embodiments, the first expression vector is a CRISPR / Cas9 expression vector. In embodiments, the first expression vector is a TALEN expression vector. In embodiments, the first expression vector is a viral expression vector. In embodiments, the viral expression vector is a lentiviral expression vector. In embodiments, the CAR is an anti-CD19 CAR.

[0143] In embodiments, the second expression vector is a retroviral expression vector.

[0144] In embodiments, the method includes activating the plurality of T cells prior to contacting the plurality of T cells with the first expression vector. In embodiments, the activating includes contacting the plurality of T cells with a CD3 protein and / or a CD28 protein. In embodiments, the activating includes contacting the plurality of T cells with a CD3 protein and a CD28 protein. In embodiments, the activating includes contacting the plurality of T cells with a CD3 protein or a CD28 protein. In embodiments, the activating includes contacting the plurality of T cells with a CD3 protein. In embodiments, the activating includes contacting the plurality of T cells with a CD28 protein.PATENT Attorney Docket No.048537-668001WO Methods of Use

[0145] The methods provided herein may be useful, inter alia, for producing an expanded population of mycophenolate resistant CAR-T cells for use in a method of treatment. In embodiments, the expanded population of mycophenolate resistant CAR-T cells are useful, inter alia, for treating cancer. using mycophenolate-resistant CAR-T cells for the treatment of cancer. In embodiments, the mycophenolate resistant CAR-T cells may be co-administered (e.g., sequentially or simultaneously) with mycophenolate as a combination therapy. In embodiments, the methods provided herein allows for immunosuppression of endogenous cells and selective support of genetically modified CAR-T cells. In embodiments, the methods provided herein prevents graft vs host disease (GVHD) and / or immune rejection. In embodiments, the pharmaceutical compositions provided herein are, inter alia, useful for treating cancer concurrently with an immunosuppressive agent (e.g., mycophenolate). In embodiments, the methods provided herein including embodiments thereof may be useful, inter alia, for improving the safety and durability of adoptive cell therapies.

[0146] Thus, in an aspect is provided a method of treating cancer in a subject in need thereof, the method including administering a therapeutically effective amount of an expanded population of mycophenolate resistant CAR-T cells produced by the methods provided herein including embodiments thereof or the pharmaceutical compositions provided herein including embodiments thereof.

[0147] In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1.5 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 2 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 2.5 x 106and about 1 x 109mycophenolate resistant CAR-T cells perPATENT Attorney Docket No.048537-668001WO kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 3 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 3.5 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 4 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 4.5 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 5 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 5.5 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 6 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 6.5 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 7 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 7.5 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 8 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-TPATENT Attorney Docket No.048537-668001WO cells includes between about 8.5 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 9 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 9.5 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight.

[0148] In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1.5 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 2 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 2.5 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 3 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 3.5 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 4 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 4.5 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 5 x 107and about 1 xPATENT Attorney Docket No.048537-668001WO 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 5.5 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 6 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 6.5 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 7 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 7.5 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 8 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 8.5 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 9 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 9.5 x 107and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight.

[0149] In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1.5 x 108and about 1 x 109mycophenolate resistant CAR-T cells perPATENT Attorney Docket No.048537-668001WO kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 2 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 2.5 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 3 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 3.5 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 4 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 4.5 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 5 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 5.5 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 6 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 6.5 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 7 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-TPATENT Attorney Docket No.048537-668001WO cells includes between about 7.5 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 8 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 8.5 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 9 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 9.5 x 108and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight.

[0150] In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 9.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 9 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 8.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 8 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 7.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 7 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 6.5 xPATENT Attorney Docket No.048537-668001WO 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 6 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 5.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 4.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 4 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 3.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 3 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 2.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 2 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 1.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 1 x 108mycophenolate resistant CAR-T cells per kilogram of body weight.PATENT Attorney Docket No.048537-668001WO

[0151] In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 9.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 9 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 8.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 8 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 7.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 7 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 6.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 6 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 5.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 4.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-TPATENT Attorney Docket No.048537-668001WO cells includes between about 1 x 106and about 4 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 3.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 3 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 2.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 2 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 1.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 1 x 107mycophenolate resistant CAR-T cells per kilogram of body weight.

[0152] In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 9.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 9 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 8.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 8 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 7.5 xPATENT Attorney Docket No.048537-668001WO 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 7 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 6.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 6 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 5.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 4.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 4 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 3.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 3 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 2.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 2 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expandedPATENT Attorney Docket No.048537-668001WO population of mycophenolate resistant CAR-T cells includes between about 1 x 106and about 1.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight.

[0153] In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 1 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 1.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 2 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 2.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 3 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 3.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 4 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 4.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 5.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 6 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 6.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolatePATENT Attorney Docket No.048537-668001WO resistant CAR-T cells about 7 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 7.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 8 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 8.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 9 x 106mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 9.5 x 106mycophenolate resistant CAR-T cells per kilogram of body weight.

[0154] In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 1 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 1.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 2 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 2.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 3 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 3.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 4 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 4.5 x 107mycophenolate resistant CAR-T cells perPATENT Attorney Docket No.048537-668001WO kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 5.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 6 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 6.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 7 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 7.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 8 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 8.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 9 x 107mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 9.5 x 107mycophenolate resistant CAR-T cells per kilogram of body weight.

[0155] In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 1 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 1.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 2 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount ofPATENT Attorney Docket No.048537-668001WO the expanded population of mycophenolate resistant CAR-T cells about 2.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 3 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 3.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 4 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 4.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 5.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 6 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 6.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 7 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 7.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 8 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 8.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 9 x 108PATENT Attorney Docket No.048537-668001WO mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 9.5 x 108mycophenolate resistant CAR-T cells per kilogram of body weight. In embodiments, the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight.

[0156] In embodiments, the method includes administering a therapeutically effective amount of mycophenolate to the subject. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 200 mg / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 300 mg / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 400 mg / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 500 mg / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 600 mg / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 700 mg / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 800 mg / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 900 mg / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 1 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 1.1 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 1.2 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 1.3 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 1.4 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 1.5 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 1.6 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is betweenPATENT Attorney Docket No.048537-668001WO about 1.7 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 1.8 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 1.9 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 2 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 2.1 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 2.2 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 2.3 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 2.4 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 2.5 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 2.6 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 2.7 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 2.8 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 2.9 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 3 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 3.1 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 3.2 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 3.3 g / day and about 3.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 3.4 g / day and about 3.5 g / day.

[0157] In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 3.4 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 3.3 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 3.2 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 3.1 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 3 g / day. In embodiments, the therapeuticallyPATENT Attorney Docket No.048537-668001WO effective amount of mycophenolate is between about 100 mg / day and about 2.9 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 2.8 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 2.7 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 2.6 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 2.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 2.4 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 2.3 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 2.2 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 2.1 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 2 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 1.9 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 1.8 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 1.7 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 1.6 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 1.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 1.4 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 1.3 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 1.2 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 1.1 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 1 g / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 900 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 800 mg / day. In embodiments, the therapeutically effective amount of mycophenolate isPATENT Attorney Docket No.048537-668001WO between about 100 mg / day and about 700 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 600 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 500 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 400 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 300 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is between about 100 mg / day and about 200 mg / day.

[0158] In embodiments, the therapeutically effective amount of mycophenolate is about 100 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is about 200 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is about 300 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is about 400 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is about 500 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is about 600 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is about 700 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is about 800 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is about 900 mg / day. In embodiments, the therapeutically effective amount of mycophenolate is about 1 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 1.1 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 1.2 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 1.3 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 1.4 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 1.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 1.6 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 1.7 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 1.8 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 1.9 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 2 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 2.1 g / day. InPATENT Attorney Docket No.048537-668001WO embodiments, the therapeutically effective amount of mycophenolate is about 2.2 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 2.3 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 2.4 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 2.5 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 2.6 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 2.7 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 2.8 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 2.9 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 3 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 3.1 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 3.2 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 3.3 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 3.4 g / day. In embodiments, the therapeutically effective amount of mycophenolate is about 3.5 g / day.

[0159] In embodiments, the expanded population of mycophenolate resistant CAR-T cells and the mycophenolate are administered simultaneously. In embodiments, the expanded population of mycophenolate resistant CAR-T cells and the mycophenolate are administered sequentially. In embodiments, the expanded population of mycophenolate resistant CAR-T cells is administered at a first time point and the mycophenolate is administered at a second time point. In embodiments, the first time point is before the second time point. In embodiments, the second time point is before the first time point.

[0160] In embodiments, the expanded population of mycophenolate resistant CAR-T cells are allogeneic to the subject.

[0161] In embodiments, the cancer is leukemia, lymphoma, myeloma, glioma, sarcoma, melanoma, carcinoma, or neuroblastoma. In embodiments, the cancer is leukemia. In embodiments, the cancer is lymphoma. In embodiments, the cancer is myeloma. In embodiments, the cancer is glioma. In embodiments, the cancer is sarcoma. In embodiments, the cancer is melanoma. In embodiments, the cancer is carcinoma. In embodiments, the cancer is neuroblastoma.PATENT Attorney Docket No.048537-668001WO

[0162] In embodiments, the cancer is B-cell acute lymphoblastic leukemia (B-ALL), T-cell acute lymphoblastic leukemia (T-ALL), chronic lymphocytic leukemia (CLL), chronic myelomonocytic leukemia (CMML), hairy cell leukemia, or blastic plasmacytoid dendritic cell neoplasm (BPDCN).

[0163] In embodiments, the cancer is B-cell acute lymphoblastic leukemia (B-ALL). In embodiments, the cancer is T-cell acute lymphoblastic leukemia (T-ALL). In embodiments, the cancer is chronic lymphocytic leukemia (CLL). In embodiments, the cancer is chronic myelomonocytic leukemia (CMML). In embodiments, the cancer is hairy cell leukemia. In embodiments, the cancer is blastic plasmacytoid dendritic cell neoplasm (BPDCN).

[0164] In embodiments, the cancer is diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), follicular lymphoma, plasmablastic lymphoma, primary mediastinal large B-cell lymphoma, or Hodgkin lymphoma. In embodiments, the cancer is diffuse large B-cell lymphoma (DLBCL). In embodiments, the cancer is mantle cell lymphoma (MCL). In embodiments, the cancer is follicular lymphoma. In embodiments, the cancer is plasmablastic lymphoma. In embodiments, the cancer is primary mediastinal large B-cell lymphoma. In embodiments, the cancer is Hodgkin lymphoma.

[0165] In embodiments, the cancer is multiple myeloma. In embodiments, the cancer is osteosarcoma, Ewing sarcoma, or rhabdomyosarcoma. In embodiments, the cancer is osteosarcoma. In embodiments, the cancer is Ewing sarcoma. In embodiments, the cancer is rhabdomyosarcoma. In embodiments, the cancer is cutaneous melanoma or uveal melanoma. In embodiments, the cancer is cutaneous melanoma. In embodiments, the cancer is uveal melanoma.

[0166] In embodiments, the cancer is non-small cell lung cancer (NSCLC), triple-negative breast cancer (TNBC), pancreatic ductal adenocarcinoma, colorectal carcinoma, ovarian carcinoma, liver cancer, or head and neck squamous cell carcinoma (HNSCC). In embodiments, the cancer is non- small cell lung cancer (NSCLC). In embodiments, the cancer is triple-negative breast cancer (TNBC). In embodiments, the cancer is pancreatic ductal adenocarcinoma. In embodiments, the cancer is colorectal carcinoma. In embodiments, the cancer is ovarian carcinoma. In embodiments,PATENT Attorney Docket No.048537-668001WO the cancer is liver cancer. In embodiments, the cancer is head and neck squamous cell carcinoma (HNSCC).

[0167] In embodiments, the cancer expresses CD19, ROR1, BCMA, CD123, or CS1 (SLAMF7). In embodiments, the cancer expresses CD19. In embodiments, the cancer expresses ROR1. In embodiments, the cancer expresses BCMA. In embodiments, the cancer expresses CD123. In embodiments, the cancer expresses CS1 (SLAMF7).

[0168] In another aspect is provided a method of treating cancer in a subject in need thereof, the method including administering to the subject a therapeutically effective amount of allogeneic CAR T-cells and an effective amount of an immune disrupting agent, wherein the allogenic CAR T-cells include a chimeric antigen receptor capable of binding to an anti-cancer antigen.

[0169] In embodiments, the allogeneic CAR T-cells do not include a functional T-cell receptor (TCR). In embodiments, the allogeneic CAR T-cells do not include a T-cell receptor (TCR). In embodiments, the allogeneic CAR T-cells have a T-cell receptor (TCR) genetically inactivated. In embodiments, the allogeneic CAR T-cells have a T-cell receptor (TCR) genetically deleted (e.g., knocked out).

[0170] In embodiments, the immune disrupting agent is an inhibitor of inosine 5’-monophosphate dehydrogenase (IMPDH).

[0171] In embodiments, the immune disrupting agent is a non-competitive inhibitor of inosine 5’- monophosphate dehydrogenase (IMPDH).

[0172] In embodiments, the immune disrupting agent is mycophenolic acid or mycophenolate. In embodiments, the immune disrupting agent is mycophenolic acid. In embodiments, the immune disrupting agent is mycophenolate.

[0173] In embodiments, the anti-cancer antigen is CD19, ROR1, BCMA, CD123 or CS1 (SLAMF7). In embodiments, the anti-cancer antigen is CD19. In embodiments, the anti-cancer antigen is ROR1. In embodiments, the anti-cancer antigen is BCMA. In embodiments, the anti- cancer antigen is CD123. In embodiments, the anti-cancer antigen is CS1 (SLAMF7).PATENT Attorney Docket No.048537-668001WO

[0174] In another aspect is provided a method of expanding a CAR T-cell population derived from cord blood, the method including culturing the CAR T-cell population in the presence of an inhibitor of inosine 5’-monophosphate dehydrogenase (IMPDH).

[0175] In embodiments, the inhibitor of inosine 5’-monophosphate dehydrogenase (IMPDH) is a non-competitive inhibitor.

[0176] In embodiments, the inhibitor of inosine 5’-monophosphate dehydrogenase (IMPDH) is mycophenolic acid or mycophenolate. In embodiments, the inhibitor of inosine 5’-monophosphate dehydrogenase (IMPDH) is mycophenolic acid. In embodiments, the inhibitor of inosine 5’- monophosphate dehydrogenase (IMPDH) is mycophenolate.

[0177] In embodiments, cells in the CAR T-cell population each include a chimeric antigen receptor capable of binding to a cancer antigen. In embodiments, the CAR T-cell population each include a chimeric antigen receptor capable of binding to a cancer antigen.

[0178] In embodiments, the anti-cancer antigen is CD19, ROR1, BCMA, CD123 or CS1 (SLAMF7). In embodiments, the anti-cancer antigen is CD19. In embodiments, the anti-cancer antigen is ROR1. In embodiments, the anti-cancer antigen is BCMA. In embodiments, the anti- cancer antigen is CD123. In embodiments, the anti-cancer antigen is CS1 (SLAMF7).

[0179] In embodiments, the CAR T-cell population does not include a functional T-cell receptor (TCR). In embodiments, the CAR T-cell population does not include a T-cell receptor (TCR). In embodiments, the CAR T-cell population has T-cell receptors (TCRs) genetically inactivated. In embodiments, the CAR T-cell population has T-cell receptors (TCRs) genetically deleted (e.g., knocked out). PHARMACEUTICAL COMPOSITIONS

[0180] The methods provided herein may be useful, inter alia, for producing a pharmaceutical composition. In embodiments, the pharmaceutic compositions provided herein including embodiments thereof are, inter alia, useful for treating a disease. In embodiments, the pharmaceutical compositions may be used in the treatment of cancer. In embodiments, the pharmaceutical compositions provided herein may be useful, inter alia, for preventing graft vs hostPATENT Attorney Docket No.048537-668001WO disease (GVHD) and / or immune rejection. In embodiments, the pharmaceutical compositions provided herein are, inter alia, useful for treating cancer concurrently with an immunosuppressive agent (e.g., mycophenolate).

[0181] Thus, in an aspect is provided a pharmaceutical composition including an expanded population of mycophenolate resistant CAR-T cells produced by the methods provided herein including embodiments thereof and a pharmaceutically acceptable excipient.

[0182] In embodiments, the pharmaceutical composition further includes an immunosuppressive agent. In embodiments, the pharmaceutical composition includes mycophenolate. EXAMPLES Example 1: Generation of Novel Allogeneic Chimeric Antigen Expressing T-Lymphocytes

[0183] T-lymphocytes (T-cells) are major components of the human immune system that target infectious agents and aberrant cell growth that can transform into malignancies. In this regard, T- cells work in concert with a wide variety of cells that together constitute the first line of defense against both human pathogens and cancer. In fact, the development of human cancers is dependent on the inability of the immune system to identify and eliminate transformed cells prior to their evolution into malignancies. Because of their central role in targeting and eliminating cancers, T- cells have long been a major focus for the development of anti-cancer therapies including the activation and targeting of these cells against both solid tumor and hematological malignances.

[0184] An area of active research is the development of technologies to redirect or augment T-cell targeting human cancers. This is generally accomplished by modifying the lymphocyte’s targeting of aberrant antigens expressed on transformed cells through the modification of the T-cell Receptor (TCR), which will redirect the lymphocyte’s activity against a chosen target on a malignant cell. Another approach is the de-novo expression in the T-cell of an artificial cancer targeting system that replaces the lymphocytes endogenous antigen binding TCR. In this system a recombinant chimeric antigen receptor (CAR)targeting moiety is linked to lymphocyte activating sequences that arePATENT Attorney Docket No.048537-668001WO triggered upon target engagement. These CAR T-cells have shown to have highly specific and potent activity when employed to treat human cancers.

[0185] Currently there are 6 approved CAR-T therapies approved for the treatment of human hematological malignancies including the CD19 targeting agents: ^Kymriah (Tisagenlecleucel)^ Yescarta (Axicabtagene Ciloleucel)^ Tecartus (Brexucabtagene Autoleucel)^ Breyanzi (Lisocabtagene Maraleucel)CD19 is a protein expressed on the surface of most B-lymphocytes and B-cell malignancies including B-cell lymphoma, B-cell acute lymphoblastic leukemia, large B-cell lymphoma, mantle cell lymphoma and chronic and small cell lymphocytic leukemia.

[0186] In addition to CD19, successful CAR-Ts have been generated against B-cell maturation antigen (BCMA) including ^Abecma (Idecabtagene Vicleucel)^ Carvykti (Ciltacabtagene Autoleucel)BCMA is found primarily on mature B lymphocytes and plasma cells and is overexpressed and activated on the malignant multiple myeloma plasma cells. To date, no CAR-T cells have been approved for the treatment of solid tumor cancers; however, there are dozens of clinical trials employing CAR-T modified lymphocytes directed against a wide variety of solid tumor cancers.

[0187] Although the molecular constructs and manufacturing processes are somewhat different for the approved products, the basic process to generate these CAR-T therapeutics are dependent on the generation of an autologous product from the patient’s own lymphocytes. This process requires the following steps including: ^Bulk lymphocyte collection by leukapheresis, which is an invasive process that isolates thewhite cells from the blood utilizing elutriation ^T-cell isolation from the leukapheresed product by magnetic bead selection^ T-cell activation and transduction employing retroviral vectors^ Expansion of the transduced product to therapeutic cell quantitiesPATENT Attorney Docket No.048537-668001WO ^Collection, formulation and fill and finish of the autologous CAR-T cells^ Limited release testing of the final cell therapy

[0188] The generation of these autologous products require the shipment to and from specialized cleanroom facilities that generate the therapeutic products. These facilities employ highly skilled production personnel who are creating these labor-intensive products under defined processes and procedures. Because each autologous product is unique and generated de novo from patient lymphocytes, the functional activity of the final CAR-T can vary greatly and in certain circumstances a therapeutic product cannot be produced. Therefore, there are many issues in the production of these autologous “one-off” CAR-T products.

[0189] To obviate many of the issues associated with the autologous product, allogeneic CAR-T offer several advantages including single bulk production processes that creates “off the shelf” therapies, lower production costs, generation of homologous products that can be thoroughly tested and released and the elimination of time and logistic constraints. Because of these advantages many companies are developing allogeneic CAR-T therapies employing several different strategies to ameliorate the challenges the product pose including graft vs host disease (GVHD) and immune rejection.

[0190] The allogeneic CAR-T products under clinical development include: ^UCART19: targets CD19 and disrupts the TCR and CD52 receptor genes,minimizing the risk of graft-versus-host disease (GVHD) and making the cells resistant to the chemotherapy drug alemtuzumab. ^ALLO-501 and ALLO-501A: identical to UCART19 but uses a refinedmanufacturing process. ^ALLO-715: targets BCMA with disruption of the TCR^ PBCAR0191:targets CD19 and disrupts the TRAC gene, eliminating the TCR^ UCART123:targets CD123, expressed on AML cells with inactivation of the TCRand CD52 genes ^UCARTCS1:targets CS1 (SLAMF7), a protein expressed on multiple myelomacells.

[0191] The products above have been shown to be active in preclinical systems. However, in clinical studies these allogeneic CAR-Ts are persistent for a very short amount of time driven by aPATENT Attorney Docket No.048537-668001WO resurgence of the host immune response (in most instances, the patients undergo lymphodepletion prior to CAR-T therapy) that in turn rejects the allogeneic cells, even with employment of the immuno-suppressive agent alemtuzumab. As a result, human clinical responses to the allogeneic CAR-T products have been muted. Therefore, graft vs host disease (addressed by TCR disruption) and equally important host vs graft disease must be addressed to generate a successful allogeneic CAR-T product.

[0192] In this disclosure, we propose a novel method to generate allogeneic CAR-Ts that offer significant advantages over currently employed allogeneic CAR-T systems. In fact, the administration of chimeric antigen receptor modified T-cells is essentially a modified lymphocyte transplant and shares many elements of often employed allogeneic hematopoietic stem cell transplants (HSCT). Like allogeneic transplants, the successful employment of CAR-T transplants requires that both graft vs host and host vs graft activities be considered. For HSCT, successful allogeneic transplants are dependent on the close match of human leukocyte antigen (HLA) antigens from the donor and recipient. Additionally, immuno-suppressive regimens have been developed to ablate any untoward activities generated against the host or transplant. Immunosuppression typically is required for the duration of the transplanted allogeneic cells.

[0193] For an allogeneic universal CAR-T product, HLA matching becomes a particularly difficult problem to surmount and would require the generation of many diverse products. However, because a CAR-T is not intended as a permanent cell source or replacement, the major issues regarding graft vs host and the allogeneic transplant can be ameliorated by genetic manipulation of the transplant and transient immuno-suppression of the host, respectively. Towards this end and to inhibit GVHD, most allogeneic CAR-T products currently in clinical trials employ T-cells that have the TCR genetically inactivated or removed. The elimination of the host immune response against the transplant has been more problematic and have primarily employed chemotherapeutic agents that target T-cells including alemtuzumab and cyclophosphamide. The use of these potent and toxic agents can have long-term impact on the host and can lead to prolonged and profound inhibition of the immune system.PATENT Attorney Docket No.048537-668001WO

[0194] In this disclosure, we plan to generate CAR-T cells that are used as a transplant to target the malignancy while employing immune suppressing agents to shield the allogeneic lymphocytes from the host. In this process as has been done for the other allogeneic products, we would first disrupt the TCR to eliminate GVHD. Then to prevent host vs graft disease, we would treat the host with an immune disrupting agent, which would arrest the host immune system while the transplant is completing the removal of the malignant target. Upon therapeutic success, the immune disrupting agent would be withdrawn and the host immune activities totally restored.

[0195] Mycophenolic acid (mycophenolate) had been used since the mid-1990s as an immunosuppressive agent to prevent organ transplant rejection. More recently, mycophenolate has been employed as a steroid sparing treatment for autoimmune diseases including lupus nephritis and treatment resistant psoriasis with fewer side effects than other immuno-suppressive agents. Of greater importance, upon withdrawal of this agent the full activity of the immune system is rapidly restored with minimal long-term impact on the treated host. Mycophenolate has also been used successfully for the treatment of acute and chronic GVHD in patients receiving hematological transplants and the availability of an oral form have facilitated the long-term use of this therapeutic.

[0196] Mycophenolic acid is a potent, reversible, non-competitive inhibitor of inosine 5’- monophosphate dehydrogenase (IMPDH), an enzyme essential for the de novo generation of the nucleoside guanosine-5’monophosphate GMP, which is an essential component of nucleic acid generation and G-coupled reaction. Since lymphocyte activation is dependent on both processes and are essential for T-cell activation, IMPDH blockade by mycophenolate results in the observed immunosuppressive activity. Of interest, almost all cells in the body are dependent on GMP production for multiple cellular processes and replication. However, when used at therapeutic concentrations, mycophenolate does not generate severe side effects even when used chronically as an immunosuppressive agent.

[0197] The differential activity of mycophenolate on lymphocytes relative to other cell types may be dependent on the existence of two isoforms of IMPDH, IMPDH1 (H1) and IMPDH2 (H2). H1 is the isoform of found primarily in all cells including lymphocytes, which also express elevated levels of H2. Mycophenolate has greater activity against H2 which may explain the differential activity ofPATENT Attorney Docket No.048537-668001WO this molecule against these immune cells. Furthermore, when compared to other cell types, lymphocytes have diminished purine salvage pathway activity and the blockade of both the H1 and H2 enzymes will greatly impact de novo purine synthesis resulting in the inability of these cells to become activated and replicate.

[0198] As described above, the successful development of a human allogeneic CAR-T cell therapeutic will be dependent on the reduction if not elimination of GVHD and HVGD generated by administration of these immunotherapies. Additionally, to produce these off-the-shelf products, we will need to generate cell banks of sufficient size to commercialize this product. Towards this end, we have expanded T-cells from adults and neonates over 40 population doublings without evidence of transformation. The ability to expand these cells to these elevated amounts would allow us to expand our candidate therapeutics from single cells to lymphocyte amounts needed to treat tens of thousands of patients.

[0199] Therefore, one method we would employ to generate an allogeneic CD19 targeting CAR would begin with a cord-blood T-cell population, which we would use recombinant methods to knock out (KO) the TCR. At the same time, these cord blood T-cells are relatively naïve in identifying foreign antigens and when used as allogeneic cell sources generate less GVHD when compared to transplants from older individuals. In the first step of this process, the T-cells would be isolated and the TCR inactivated using processes used to generate the allogeneic cells currently being tested in human studies. The KO cells would then be transduced to express both the H2 isoenzyme and the CAR expression cassette that would employ a CD19 or ROR1 targeting moiety. The transduced cells expressing both H2 and the CAR product would then be expanded to amounts necessary to generate thousands of therapeutic products (determined for example by employing 1e8 cells / kg or maximally 1e10 cells / dose and minimally 1e4 products would require 1e14 cells or approximately 42 cell doublings from a single cell). Towards this end, we have previously expanded human primary T-cells over 40 cell doublings without evidence of exhaustion or transformation.

[0200] We have already demonstrated the ability to make T-cells transduced with H2 more resistant to mycophenolate. For these initial proof-of-concept studies, we created a lentiviral vector that expressed both H2 and red fluorescent protein to track the transduced cells. As shown in FIG.1PATENT Attorney Docket No.048537-668001WO using, human Jurkat T-cells transduced with the bicistronic vector, showed a dose dependent resistance to mycophenolate at concentrations that correspond to serum levels found in patients with adequate immunosuppression. More importantly, the transduced cells become more resistant to mycophenolate as the cell population becomes further selected and we would expect that this resistance would select cell populations that have even greater resistance to the toxin.

[0201] In conclusion, we have completed the initial proof-of-concept studies in generating mycophenolate resistant T-cells that will be employed for the creation of an allogeneic CAR-T product. For our next studies, we will employ human T-cells and transduce them with a bicistronic vector expressing H2 and CD19 or ROR1 modified T-cell receptor. Example 2

[0202] T-cells are isolated directly from donor fetal cord blood using StraightFrom MicroBeads (Miltenyi Biotec) per standard protocol. Automated separation is carried out to isolate CD4, CD8, and / or CD3 T-cell sub-populations using the autoMACS Pro Separator (Miltenyi Biotec). T cells are cultured in ImmunoCult™-XF T Cell Expansion Medium (Stemcell Technologies #10981, Vancouver, Canada) supplemented with Pen / Strep (Gibco), gentamycin (VWR, Radnor, PA), and IL-7 and IL-15 (25 ng / ml each-Miltenyi Biotec) in G-Rex6 or G-rex100 (Wilson Wolf, New Brighton, MN). T Cell TransAct (Miltenyi Biotec) is added per standard protocol for T-cell stimulation.1-3 days after activation, CAR lentiviruses expressing the CD19 targeting FMC63 binding moiety are added at an MOI of 5 with 10 ^g / mL protamine sulfate (Sigma-Aldrich, St Louis, MO) as a transducing agent. Media is exchanged with the standard culture media and replenished every 3-4 days during expansion.

[0203] The CAR-T cells generated are then transduced with a lentivirus that expresses the modified mycophenolate resistant IMPDH2 construct and finally the T-cell receptor (TCR) are knocked out by CRISPR or TALEN endonucleases.PATENT Attorney Docket No.048537-668001WO Example 3: CD3⁺ T Cell Isolation

[0204] T cells are isolated directly from anticoagulated whole blood using StraightFrom™ Whole Blood CD3 MicroBeads (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer’s protocol. Whole blood is collected in tubes containing EDTA, ACD-A, or CPD as anticoagulants. The blood sample is passed through a 30 µm nylon mesh filter (Miltenyi Biotec #130-041-407) to remove cell clumps prior to labeling.

[0205] Magnetic labeling is performed by adding 50 µL of StraightFrom CD3 MicroBeads per 1 mL of whole blood and incubating for 15 minutes at 2–8 °C. Labeled cells are optionally washed with separation buffer (PBS containing 0.5% BSA and 2 mM EDTA) and centrifuged at 445×g for 10 minutes. The cell pellet is resuspended in separation buffer to a final volume of 1 mL.

[0206] Magnetic separation is performed using the autoMACS® Pro Separator (Miltenyi Biotec). The labeled sample is loaded into the Chill Rack and processed using the “Posselwb” program to collect the CD3⁺ T cell fraction. The positive fraction is collected in row C of the tube rack. Alternatively, separation is performed using the MultiMACS™ Cell24 Separator Plus or the Whole Blood Column Kit (Miltenyi Biotec) with appropriate buffer volumes and elution steps as described in the manufacturer’s protocol.

[0207] The resulting CD3⁺ T cells are used for downstream methods of producing an expanded population of mycophenolate resistant CAR-T cells as described herein. Example 3: T Cell Activation and Expansion

[0208] ^Isolated T cells are activated using T Cell TransAct, human (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer’s protocol. T cells are resuspended in TexMACS™ Medium (Miltenyi Biotec #130-097-196) supplemented with 20 IU / mL Human IL-2 (Miltenyi Biotec #130-097-744) or, alternatively, with 155 U / mL Human IL-7 (Miltenyi Biotec #130-095-361) and 290 U / mL Human IL-15 (Miltenyi Biotec #130-095-762).

[0209] For activation in a 48-well plate, 1×10⁶ purified T cells are resuspended in 990 µL of supplemented TexMACS Medium.10 µL of T Cell TransAct is added to each well, and the cells arePATENT Attorney Docket No.048537-668001WO incubated at 37 °C in a humidified incubator with 5% CO₂ for up to 3 days. Cultures are inspected daily, and fresh medium is added as needed.

[0210] Residual reagent is removed 2–3 days after initial activation by replacing 900 µL of supernatant with fresh supplemented TexMACS Medium or by centrifugation at 300×g for 10 minutes followed by complete aspiration of the supernatant.1 mL of fresh supplemented TexMACS Medium is then added, and the cells are returned to incubation at 37 °C, 5% CO₂.

[0211] T cell expansion is carried out by splitting the culture every 2 days into two equal parts and adding fresh supplemented TexMACS Medium. Cultures are monitored daily and may be split more or less frequently depending on cell density. After 14 days, the activated and expanded T cells are used for downstream applications including methods of producing an expanded population of mycophenolate resistant CAR-T cells as described herein.

[0212] Optionally, the T cells are restimulated by reapplying T Cell TransAct at a reduced titer of 1:500 to support further expansion. P EMBODIMENTS

[0213] P Embodiment 1. A method of treating cancer in a subject in need thereof, the method comprising administering to said subject a therapeutically effective amount of allogeneic CAR T- cells and an effective amount of an immune disrupting agent, wherein said allogenic CAR T-cells comprise a chimeric antigen receptor capable of binding to an anti-cancer antigen.

[0214] P Embodiment 2. The method of P embodiment 1, wherein said allogeneic CAR T-cells do not comprise a functional T-cell receptor (TCR).

[0215] P Embodiment 3. The method of P embodiment 1 or 2, wherein said immune disrupting agent is an inhibitor of inosine 5’-monophosphate dehydrogenase (IMPDH).

[0216] P Embodiment 4. The method of P embodiment 1 or 2, wherein said immune disrupting agent is a non-competitive inhibitor of inosine 5’-monophosphate dehydrogenase (IMPDH).

[0217] P Embodiment 5. The method of P embodiment 1 or 2, wherein said immune disrupting agent is mycophenolic acid or mycophenolate.PATENT Attorney Docket No.048537-668001WO

[0218] P Embodiment 6. The method of one of P embodiments 1 to 5, wherein said anti-cancer antigen is CD19, ROR1, BCMA, CD123 or CS1 (SLAMF7).

[0219] P Embodiment 7. A method of expanding a CAR T-cell population derived from cord blood, the method comprising culturing said CAR T-cell population in the presence of an inhibitor of inosine 5’-monophosphate dehydrogenase (IMPDH).

[0220] P Embodiment 8. The method of P embodiment 7, wherein said inhibitor of inosine 5’- monophosphate dehydrogenase (IMPDH) is a non-competitive inhibitor.

[0221] P Embodiment 9. The method of P embodiment 7, wherein said inhibitor of inosine 5’- monophosphate dehydrogenase (IMPDH) is mycophenolic acid or mycophenolate.

[0222] P Embodiment 10. The method of one of P embodiments 7 to 9, wherein cells in said CAR T-cell population each comprise a chimeric antigen receptor capable of binding to a cancer antigen.

[0223] P Embodiment 11. The method of P embodiment 10, wherein said anti-cancer antigen is CD19, ROR1, BCMA, CD123 or CS1 (SLAMF7).

[0224] P Embodiment 12. The method of one of P embodiments 7 to 11, wherein said CAR T- cell population does not comprise a functional T-cell receptor. EMBODIMENTS

[0225] Embodiment 1. A method of producing an expanded population of mycophenolate resistant CAR-T cells, the method comprising: (i) contacting a plurality of T cells with a first expression vector comprising a first nucleic acid sequence encoding a chimeric antigen receptor (CAR) thereby producing a plurality of CAR-T cells and expanding said plurality of CAR-T cells for a first period of time thereby producing an expanded population of CAR-T cells, wherein the first period of time is at least 14 days; (ii) contacting said expanded population of CAR-T cells with a second expression vector comprising a second nucleic acid encoding a mutated IMPDH2 protein thereby producing a plurality of IMPDH2 mutant CAR-T cells, wherein said mutated IMPDH2 protein confers mycophenolate resistance to said plurality of IMPDH2 mutant CAR-T cells andPATENT Attorney Docket No.048537-668001WO expanding said plurality of IMPDH2 mutant CAR-T cells in the presence of mycophenolate for a second period of time thereby producing an expanded population of IMPDH2 CAR-T cells, wherein the second period of time is at least 14 days; and (iii) genetically altering said expanded population of IMPDH2 mutant CAR-T cells to inactivate a T-cell receptor (TCR) gene within said IMPDH2 mutant CAR-T cells thereby producing a plurality of TCR knockout IMPDH2 mutant CAR-T cells and expanding said plurality of TCR knockout IMPDH2 mutant CAR-T cells for a third period of time thereby producing an expanded population of mycophenolate resistant CAR-T cells, wherein the third period of time is at least 2 days.

[0226] Embodiment 2. The method of embodiment 1, wherein the plurality of T cells is isolated from cord blood prior to step (i).

[0227] Embodiment 3. The method of embodiment 1 or 2, wherein the expanded population of mycophenolate resistant CAR-T cells is at least 10 times more cells than the expanded population of IMPDH2 mutant CAR-T cells.

[0228] Embodiment 4. The method of any one of embodiments 1-3, wherein the expanded population of mycophenolate resistant CAR-T cells is at least 20 times more cells than the expanded population of IMPDH2 mutant CAR-T cells.

[0229] Embodiment 5. The method of any one of embodiments 1-4, wherein the expanded population of mycophenolate resistant CAR-T cells is at least 40 times more cells than the expanded population of IMPDH2 mutant CAR-T cells.

[0230] Embodiment 6. The method of any one of embodiments 1-5, wherein the expanded population of mycophenolate resistant CAR-T cells comprises between about 1 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells.

[0231] Embodiment 7. The method of any one of embodiments 1-6, wherein the first period of time is for at least 4 weeks.

[0232] Embodiment 8. The method of any one of embodiments 1-7, wherein the second period of time is for at least 4 weeks.PATENT Attorney Docket No.048537-668001WO

[0233] Embodiment 9. The method of any one of embodiments 1-8, wherein the third period of time is between about 2 days and about 60 days..

[0234] Embodiment 10. The method of any one of embodiments 1-7, wherein the mutated IMPDH2 protein comprises a catalytic site mutation.

[0235] Embodiment 11. The method of any one of embodiments 1-10, wherein the mutated IMPDH2 protein comprises an isoleucine substitution at a position corresponding to position 333 of SEQ ID NO:1 and / or a tyrosine substitution at a position corresponding to position 351 of SEQ ID NO:1.

[0236] Embodiment 12. The method of any one of embodiments 1-11, wherein the expanding of the plurality of TCR knockout IMPDH2 mutant CAR-T cells occurs in the presence of mycophenolate.

[0237] Embodiment 13. The method of any one of embodiments 1-12, wherein the first expression vector is a CRISPR / Cas9 expression vector, a TALEN expression vector, or a viral expression vector.

[0238] Embodiment 14. The method of embodiment 13, wherein the viral expression vector is a lentiviral expression vector.

[0239] Embodiment 15. The method of any one of embodiments 1-14, wherein the CAR is an anti-CD19 CAR.

[0240] Embodiment 16. The method of any one of embodiments 1-15, wherein the second expression vector is a retroviral expression vector.

[0241] Embodiment 17. The method of any one of embodiments 1-16, further comprising activating the plurality of T cells prior to contacting the plurality of T cells with the first expression vector, wherein the activating includes contacting the plurality of T cells with a CD3 protein and / or a CD28 protein.PATENT Attorney Docket No.048537-668001WO

[0242] Embodiment 18. A pharmaceutical composition comprising an expanded population of mycophenolate resistant CAR-T cells produced by the method of any one of embodiments 1-17 and a pharmaceutically acceptable excipient.

[0243] Embodiment 19. A method of treating cancer in a subject in need thereof, the method comprising administering a therapeutically effective amount of an expanded population of mycophenolate resistant CAR-T cells produced by the method of any one of embodiments 1-17 or the pharmaceutical composition of embodiment 18.

[0244] Embodiment 20. The method of embodiment 19, wherein the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells comprises between about 1 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight.

[0245] Embodiment 21. The method of embodiment 19 or 20, further comprising administering a therapeutically effective amount of mycophenolate to the subject.

[0246] Embodiment 22. The method of embodiment 21, wherein the expanded population of mycophenolate resistant CAR-T cells and the mycophenolate are administered simultaneously.

[0247] Embodiment 23. The method of embodiment 21, wherein the expanded population of mycophenolate resistant CAR-T cells and the mycophenolate are administered sequentially.

[0248] Embodiment 24. The method of any one of embodiments 19-23, wherein the expanded population of mycophenolate resistant CAR-T cells are allogeneic to the subject. INFORMAL SEQUENCE LISTING

[0249] SEQ ID NO:1 - IMPDH2 sequence isoform predominantly found in activated lymphocytes MADYLISGGTSYVPDDGLTAQQLFNCGDGLTYNDFLILPGYIDFTADQVDLTSALTKKITLKTPLVS SPMDTVTEAGMAIAMALTGGIGFIHHNCTPEFQANEVRKVKKYEQGFITDPVVLSPKDRVRDVFEAK ARHGFCGIPITDTGRMGSRLVGIISSRDIDFLKEEEHDCFLEEIMTKREDLVVAPAGITLKEANEIL QRSKKGKLPIVNEDDELVAIIARTDLKKNRDYPLASKDAKKQLLCGAAIGTHEDDKYRLDLLAQAGVPATENT Attorney Docket No.048537-668001WO DVVVLDSSQGNSIFQINMIKYIKDKYPNLQVIGGNVVTAAQAKNLIDAGVDALRVGMGSGSICITQE VLACGRPQATAVYKVSEYARRFGVPVIADGGIQNVGHIAKALALGASTVMMGSLLAATTEAPGEYFF SDGIRLKKYRGMGSLDAMDKHLSSQNRYFSEADKIKVAQGVSGAVQDKGSIHKFVPYLIAGIQHSCQ DIGAKSLTQVRAMMYSGELKFEKRTSSAQVEGGVHSLHSYEKRLF

[0250] SEQ ID NO:2 - IMPDH2 mycophenolate-resistant mutant MADYLISGGTSYVPDDGLTAQQLFNCGDGLTYNDFLILPGYIDFTADQVDLTSALTKKITLKTPLVS SPMDTVTEAGMAIAMALTGGIGFIHHNCTPEFQANEVRKVKKYEQGFITDPVVLSPKDRVRDVFEAK ARHGFCGIPITDTGRMGSRLVGIISSRDIDFLKEEEHDCFLEEIMTKREDLVVAPAGITLKEANEIL QRSKKGKLPIVNEDDELVAIIARTDLKKNRDYPLASKDAKKQLLCGAAIGTHEDDKYRLDLLAQAGV DVVVLDSSQGNSIFQINMIKYIKDKYPNLQVIGGNVVTAAQAKNLIDAGVDALRVGMGSGSICIIQE VLACGRPQATAVYKVYEYARRFGVPVIADGGIQNVGHIAKALALGASTVMMGSLLAATTEAPGEYFF SDGIRLKKYRGMGSLDAMDKHLSSQNRYFSEADKIKVAQGVSGAVQDKGSIHKFVPYLIAGIQHSCQ DIGAKSLTQVRAMMYSGELKFEKRTSSAQVEGGVHSLHSYEKRLF

[0251] SEQ ID NO:3 - IMPDH alternative isoform MADYLISGGTSYVPDDGLTAQQLFNCGDGLTYNDFLILPGYIDFTADQVDLTSALTKKITLKTPLVS SPMDTVTEAGMAIAMALTGGIGFIHHNCTPEFQANEVRKVKKYEQGFITDPVVLSPKDRVRDVFEAK ARHGFCGIPITDTGRMGSRLVGIISSRDIDFLKEEEHDCFLEEIMTKREDLVVAPAGITLKEANEIL QRSKKGKLPIVNEDDELVAIIARTDLKKNRDYPLASKDAKKQLLCGAAIGTHEDDKYRLDLLAQAGV DVVVLDSSQGNSIFQINMIKYIKDKYPNLQVIGGNVVTAAQAKNLIDAGVDALRVGMGSGSICITQE VAPKIPPDIKSHSPKCPSTVTGCYMLACGRPQATAVYKVSEYARRFGVPVIADGGIQNVGHIAKALA LGASTVMMGSLLAATTEAPGEYFFSDGIRLKKYRGMGSLDAMDKHLSSQNRYFSEADKIKVAQGVSG AVQDKGSIHKFVPYLIAGIQHSCQDIGAKSLTQVRAMMYSGELKFEKRTSSAQVEGGVHSLHSYEKR LF

Claims

PATENT Attorney Docket No.048537-668001WO WHAT IS CLAIMED IS:

1. A method of producing an expanded population of mycophenolate resistant CAR-T cells, the method comprising: (i) contacting a plurality of T cells with a first expression vector comprising a first nucleic acid sequence encoding a chimeric antigen receptor (CAR) thereby producing a plurality of CAR-T cells and expanding said plurality of CAR-T cells for a first period of time thereby producing an expanded population of CAR-T cells, wherein the first period of time is at least 14 days; (ii) contacting said expanded population of CAR-T cells with a second expression vector comprising a second nucleic acid encoding a mutated IMPDH2 protein thereby producing a plurality of IMPDH2 mutant CAR-T cells, wherein said mutated IMPDH2 protein confers mycophenolate resistance to said plurality of IMPDH2 mutant CAR-T cells and expanding said plurality of IMPDH2 mutant CAR-T cells in the presence of mycophenolate for a second period of time thereby producing an expanded population of IMPDH2 CAR-T cells, wherein the second period of time is at least 14 days; and (iii) genetically altering said expanded population of IMPDH2 mutant CAR-T cells to inactivate a T-cell receptor (TCR) gene within said IMPDH2 mutant CAR-T cells thereby producing a plurality of TCR knockout IMPDH2 mutant CAR-T cells and expanding said plurality of TCR knockout IMPDH2 mutant CAR-T cells for a third period of time thereby producing an expanded population of mycophenolate resistant CAR-T cells, wherein the third period of time is at least 2 days.

2. The method of claim 1, wherein the plurality of T cells is isolated from cord blood prior to step (i).

3. The method of claim 1, wherein the expanded population of mycophenolate resistant CAR-T cells is at least 10 times more cells than the expanded population of IMPDH2 mutant CAR-T cells.

4. The method of claim 1, wherein the expanded population of mycophenolate resistant CAR-T cells is at least 20 times more cells than the expanded population of IMPDH2 mutant CAR-T cells.PATENT Attorney Docket No.048537-668P01US Client Ref. No. SD2025-062-1 5. The method of claim 1, wherein the expanded population of mycophenolate resistant CAR-T cells is at least 40 times more cells than the expanded population of IMPDH2 mutant CAR-T cells.

6. The method of claim 1, wherein the expanded population of mycophenolate resistant CAR-T cells comprises between about 1 x 1012and about 8 x 1014mycophenolate resistant CAR-T cells.

7. The method of claim 1, wherein the first period of time is for at least 4 weeks.

8. The method of claim 1, wherein the second period of time is for at least 4 weeks.

9. The method of claim 1, wherein the third period of time is between about 2 days and about 60 days.

10. The method of claim 1, wherein the mutated IMPDH2 protein comprises a catalytic site mutation.

11. The method of claim 1, wherein the mutated IMPDH2 protein comprises an isoleucine substitution at a position corresponding to position 333 of SEQ ID NO:1 and / or a tyrosine substitution at a position corresponding to position 351 of SEQ ID NO:

1.

12. The method of claim 1, wherein the expanding of the plurality of TCR knockout IMPDH2 mutant CAR-T cells occurs in the presence of mycophenolate.

13. The method of claim 1, wherein the first expression vector is a CRISPR / Cas9 expression vector, a TALEN expression vector, or a viral expression vector.

14. The method of claim 13, wherein the viral expression vector is a lentiviral expression vector.

15. The method of claim 1, wherein the CAR is an anti-CD19 CAR.PATENT Attorney Docket No.048537-668P01US Client Ref. No. SD2025-062-1 16. The method of claim 1, wherein the second expression vector is a retroviral expression vector.

17. The method of claim 1, further comprising activating the plurality of T cells prior to contacting the plurality of T cells with the first expression vector, wherein the activating includes contacting the plurality of T cells with a CD3 protein and / or a CD28 protein.

18. A pharmaceutical composition comprising an expanded population of mycophenolate resistant CAR-T cells produced by the method of claim 1 and a pharmaceutically acceptable excipient.

19. A method of treating cancer in a subject in need thereof, the method comprising administering a therapeutically effective amount of an expanded population of mycophenolate resistant CAR-T cells produced by the method of claim 1 or the pharmaceutical composition of claim 18.

20. The method of claim 19, wherein the therapeutically effective amount of the expanded population of mycophenolate resistant CAR-T cells comprises between about 1 x 106and about 1 x 109mycophenolate resistant CAR-T cells per kilogram of body weight.

21. The method of claim 19, further comprising administering a therapeutically effective amount of mycophenolate to the subject.

22. The method of claim 21, wherein the expanded population of mycophenolate resistant CAR-T cells and the mycophenolate are administered simultaneously.

23. The method of claim 21, wherein the expanded population of mycophenolate resistant CAR-T cells and the mycophenolate are administered sequentially.

24. The method of claim 19, wherein the expanded population of mycophenolate resistant CAR-T cells are allogeneic to the subject.

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