Immunotherapeutic methods and compositions involving tryptophan metabolic pathway modulators - Patents.com

Abstract

Therapeutic methods involving immunotherapy (e.g., T cell therapy) and administration of a tryptophan metabolism and / or kynurenine pathway modulator are provided. In some embodiments, the methods include combination therapy involving administration of engineered T cells (e.g., chimeric antigen receptor (CAR)-expressing cells) and a tryptophan metabolism and / or kynurenine pathway modulator (e.g., an enzyme inhibitor). Engineered cells in which the expression of molecules involved in the kynurenine pathway is altered are also provided. Methods for producing engineered cells for use in these methods, cells, compositions, methods for administering to subjects, nucleic acids, and kits are also provided. In some aspects, the features of the methods and cells of the present invention result in increased or improved activity, activity, outcome, function, response, persistence, expansion, and / or proliferation of cells for adoptive cell therapy.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 62 / 407,776, filed October 13, 2016, entitled "Immunotherapeutic Methods and Compositions Comprising Tryptophan Metabolic Pathway Modulators," and U.S. Provisional Patent Application No. 62 / 514,767, filed June 2, 2017, entitled "Immunotherapeutic Methods and Compositions Comprising Tryptophan Metabolic Pathway Modulators," the entire contents of which are incorporated herein by reference.

[0002] INCORPORATION-BY-REFERENCE TO SEQUENCE LISTING This application has been submitted with an electronic Sequence Listing, which is provided as a file entitled 735042007540SeqList.txt, created on October 12, 2017, and having a size of 136 kilobytes. The information in the electronic format of the Sequence Listing is incorporated by reference in its entirety.

[0003] Field The present disclosure relates in some aspects to methods, compositions, and uses, including immunotherapies such as adoptive cell therapy, e.g., T cell therapy, as well as modulators of pathways involved in tryptophan metabolism, such as alternative tryptophan metabolic pathways, and / or components or metabolites thereof. In some embodiments, the modulators include kynurenine pathway modulators, including modulators of one or more reactions or components that promote tryptophan metabolism via the kynurenine pathway, and / or metabolites thereof. Provided methods, compositions, and uses include those for combination therapy, including the administration or use of one or more such modulators (e.g., inhibitors of enzymes in such pathway(s)) in combination with another agent, e.g., an immunotherapeutic agent, e.g., a therapeutic antibody, e.g., a multispecific (e.g., T cell-binding) antibody, and / or a cell therapy, e.g., a chimeric antigen receptor (CAR)-expressing T cell. Also provided are compositions and uses for one or more such immunotherapeutics, such as engineered cells that have been treated, modified, or engineered with or to contain one or more such modulators. Additionally, some engineered cells have modified, altered, disrupted, or affected expression, activity, or function of molecules, such as proteins, involved in the immunosuppressive activity of cells in response to tryptophan metabolism, such as tryptophan catabolism. Also provided are methods for producing engineered cells, cells for use in the methods, compositions, methods of administration to a subject, nucleic acids, articles of manufacture, and kits. In some embodiments, combinations are provided that include an immunotherapeutic agent, such as engineered cells, a kynurenine and / or tryptophan modulating agent, and instructions for their administration. In some aspects, features of the methods and cells provide for increased or improved activity, function, persistence, expansion, and / or proliferation of endogenous immune cells recruited by cells or immunotherapeutic agents for adoptive cell therapy. [Background technology]

[0004] background Various strategies are available for immunotherapy, for example, for adoptive therapy, engineered T cells can be administered.For example, a strategy is available for engineering T cells that express engineered antigen receptors, such as CAR, and administering compositions containing such cells to subjects.There is a need for improved strategies for cell therapy.For example, in some situations, it is desirable to have a strategy that involves improving cells or outcomes after administration in one or more aspects, for example, improving the persistence, activity and / or proliferation of cells when administered to subjects.Methods, cells, compositions, kits and systems that meet such needs are provided in some embodiments. Summary of the Invention

[0005] overview Provided herein are methods for enhancing or modulating the proliferation and / or activity of immune cell activity in immunotherapy, such as T cell therapy (e.g., CAR-expressing T cells) or T cell-binding therapy. In some embodiments, the methods generally involve administering a combination therapy of an immunotherapy, such as T cell therapy (e.g., CAR-expressing T cells) or T cell-binding therapy, with a tryptophan metabolism and / or kynurenine pathway modulator.

[0006] Provided herein are methods of treatment that include: (a) administering a T cell therapy to a subject having a disease or condition; and (b) administering a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator to the subject, wherein administration of the tryptophan metabolism and / or kynurenine pathway modulator is initiated more than one day prior to the initiation of administration of the T cell therapy.

[0007] Provided herein are methods of treatment that include administering T cell therapy to a subject having a disease or condition who has previously been administered a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator starting more than one day prior to the initiation of the T cell therapy.

[0008] In some embodiments of any of the methods provided herein, administration of the tryptophan metabolism and / or kynurenine pathway modulator is initiated within, or within about, 2, 3, 6, 12, 15, 30, 60, or 90 days or more before initiation of administration of T cell therapy. In some embodiments, the method further comprises continuing administration of the tryptophan metabolism and / or kynurenine pathway modulator concurrently with or after initiation of administration of T cell therapy. In some embodiments, the method further comprises administering the tryptophan metabolism and / or kynurenine pathway modulator following administration of the therapeutic cell therapy.

[0009] Provided herein are therapeutic methods comprising: (a) administering T cell therapy to a subject having a disease or condition; and (b) administering a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator to the subject after initiation of administration of T cell therapy. Also provided are therapeutic methods comprising administering a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator to a subject having cancer who has previously been administered T cell therapy. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered within, or about, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, or 48 hours after initiation of T cell therapy.

[0010] In some embodiments, prior to initiation of administration of the composition comprising the T cells, the subject has failed to respond or not fully responded to treatment with prior administration of a tryptophan metabolism and / or kynurenine pathway modulator, has relapsed after remission following treatment, or has become refractory to treatment, optionally in combination with an immunomodulatory agent, optionally in combination with a checkpoint inhibitor.

[0011] Provided herein are methods of treatment comprising: (a) administering a T cell therapy to a subject having a disease or condition; and (b) administering a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator to the subject, wherein the subject has failed to respond or has not fully responded to treatment with prior administration of the tryptophan metabolism and / or kynurenine pathway modulator, has relapsed after remission following treatment, or has become refractory to treatment, prior to initiation of administration of the composition comprising T cells, and optionally such prior administration was in combination with an immunomodulatory agent, optionally in combination with a checkpoint inhibitor.

[0012] Provided herein are methods of treatment comprising administering a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator to a subject having a disease or condition who has previously been administered T cell therapy, wherein the subject, prior to initiation of administration of the composition comprising T cells, did not respond or did not fully respond to treatment with prior administration of the tryptophan metabolism and / or kynurenine pathway modulator, relapsed after remission following treatment, or became refractory to treatment, and wherein such prior administration was optionally in combination with an immunomodulator, optionally in combination with a checkpoint inhibitor.

[0013] In some embodiments, the disease or condition is cancer.

[0014] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator administered in (b) is administered prior to, concurrently with, and / or after initiation of administration of T cell therapy. In some embodiments, administration of the tryptophan metabolism and / or kynurenine pathway modulator administered in (b) is initiated within, or within about, 2, 3, 6, 12, 15, 30, 60, or 90 days or more before initiation of administration of T cell therapy.

[0015] In some embodiments, the method also includes administering a tryptophan metabolism and / or kynurenine pathway modulator administered in (b) following initiation of administration of the T cell therapy.

[0016] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered at the following time points: when there is a decrease in tryptophan levels, an increase in kynurenine levels, and / or an increase in IDO1, IDO2, or TDO expression or activity in a biological sample from the subject compared to before administration of T cell therapy or compared to an earlier time point after administration of T cell therapy; before the number of T cell therapy cells detectable in the blood from the subject is decreased compared to in the subject at an earlier time point after administration of T cell therapy is initiated; before T cells exhibit prolonged tryptophan starvation, depletion, decreased effector function, expression of inhibitory receptors, and / or loss of proliferative capacity; before peak or maximum levels of T cell therapy cells are detectable in the subject's blood; and / or when levels of interferon-gamma (IFNγ) are increased in a biological sample from the subject compared to before administration of T cell therapy or compared to an earlier time point after administration of T cell therapy is initiated. In some embodiments, the increase or decrease is greater than 1.2-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, or more, or greater than about 1.2-fold, about 1.5-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 10-fold, or more.

[0017] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator administered in (b) is administered within 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 5 days, 6 days, 7 days, 14 days, 21 days, 28 days, or within about said time period after initiation of T cell therapy.

[0018] In some embodiments of any of the methods provided herein, the tryptophan metabolism and / or kynurenine pathway modulator is administered at a time point where there is a decrease in tryptophan levels, an increase in kynurenine levels, and / or an increase in IDO1, IDO2, or TDO expression or activity in a biological sample from the subject compared to before initiation of administration of T cell therapy or compared to an earlier time point after initiation of administration of T cell therapy; a decrease in the number of T cell therapy cells detectable in the blood from the subject compared to in the subject at an earlier time point after initiation of administration of T cell therapy; a decrease in the number of T cell therapy cells detectable in the blood compared to the peak or maximum number of T cell therapy cells detectable in the blood of the subject after initiation of administration of T cell therapy. or more than about a 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 50-fold, or 100-fold or more decrease in T cell therapy cells; and / or at a time after the peak or maximum level of T cell therapy cells is detectable in the subject's blood, the number of T cell therapy cells detectable in the subject's blood is less than 10%, less than 5%, less than 1%, or less than 0.1% of the total peripheral blood mononuclear cells (PBMCs) in the subject's blood; or at a time when the level of IFN-γ is increased in a biological sample from the subject compared to before administration of T cell therapy is initiated or compared to an earlier time point after administration of T cell therapy is initiated. In some embodiments, the increase or decrease is greater than or about 1.2-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold or more. In some embodiments, the biological sample is serum or plasma or a tumor sample, or is a tumor.

[0019] In some embodiments of any of the methods provided herein, the tryptophan metabolism and / or kynurenine pathway modulator prevents or reduces the formation, concentration, availability, metabolism, and / or action of kynurenine pathway enzymes and / or metabolites. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator prevents or reduces the catabolism of tryptophan, or prevents or reduces the synthesis or accumulation of tryptophan metabolites. In some embodiments, the tryptophan metabolite is selected from kynurenine, 3-hydroxykynurenine, and 3-hydroxyanthranilic acid (3-HAA). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a small molecule, peptide, protein, antibody or antigen-binding fragment thereof, antibody mimetic, aptamer, or nucleic acid molecule.

[0020] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is an antagonist of the enzyme kynureninase.

[0021] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of one or more enzymes selected from the group consisting of inhibitors of indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1), IDO2, and inhibitors of tryptophan 2,3-dioxygenase (TDO). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a dual-acting IDO / TDO inhibitor. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1).

[0022] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of IDO1, and the tryptophan metabolism and / or kynurenine pathway modulator of IDO1 is administered at a time when there is increased expression or activity of IDO1 in a biological sample from the subject compared to before administration of T cell therapy is initiated or compared to an earlier time point after administration of T cell therapy is initiated.

[0023] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is tryptophan.

[0024] In some embodiments of any of the methods provided herein, the cancer comprises an IDO1-negative tumor, and / or the subject has not been selected for the development of an IDO1-positive tumor; and / or the cancer comprises a TDO-negative tumor, and / or the subject has not been selected for the development of a TDO-positive tumor. In some embodiments, the cancer comprises an IDO1-negative tumor, and / or the subject has not been selected for the development of an IDO1-positive tumor.

[0025] In some embodiments of any of the methods provided herein, the cancer comprises an IDO1-negative tumor prior to initiation of administration of T cell therapy, and / or the subject has not been selected for the development of an IDO1-positive tumor prior to initiation of administration of T cell therapy; and / or the cancer comprises a TDO-negative tumor prior to initiation of administration of T cell therapy, and / or the subject has not been selected for the development of a TDO-positive tumor prior to initiation of administration of T cell therapy. In some embodiments, the cancer comprises an IDO1-negative tumor prior to initiation of administration of T cell therapy, and / or the subject has not been selected for the development of an IDO1-positive tumor prior to initiation of administration of T cell therapy.

[0026] Provided herein are methods of treatment that include: (a) administering T cell therapy to a subject having a disease or condition, wherein the cancer comprises a tumor that is negative for indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1) or tryptophan 2,3-dioxygenase (TDO) prior to initiation of administration of the T cell therapy, and / or the subject has not been selected for the development of an IDO1-positive tumor or a TDO-positive tumor prior to initiation of administration of the T cell therapy; and (b) administering to the subject a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator that is an inhibitor of IDO1.

[0027] Also provided herein are methods of treatment that include administering a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator that is an inhibitor of IDO1 to a subject having cancer who has been administered T cell therapy, wherein the cancer comprises a tumor that is negative for indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1) or tryptophan 2,3-dioxygenase (TDO) prior to initiation of administration of T cell therapy, and / or the subject has not been selected for the development of an IDO1-positive tumor or a TDO-positive tumor prior to initiation of administration of T cell therapy.

[0028] Also provided herein are methods of treatment that include administering T cell therapy to a subject having cancer who has previously been administered a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator that is an inhibitor of IDO1, wherein the cancer comprises tumors that are negative for indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1) or tryptophan 2,3-dioxygenase (TDO) prior to initiation of administration of the T cell therapy, and / or the subject has not been selected for the development of IDO1-positive tumors or TDO-positive tumors prior to initiation of administration of the T cell therapy.

[0029] In some embodiments, the disease or condition is cancer.

[0030] In some embodiments of any of the methods provided herein, the tryptophan metabolism and / or kynurenine pathway modulator is administered prior to, concurrently with, and / or after initiation of administration of T cell therapy.

[0031] In some embodiments, prior to initiation of administration of the composition comprising the T cells, the subject has failed or incompletely responded to treatment with prior administration of a tryptophan metabolism and / or kynurenine pathway modulator, has relapsed after remission following treatment, or has become refractory to treatment, where such prior administration was optionally in combination with an immunomodulatory agent, optionally in combination with a checkpoint inhibitor.

[0032] In some embodiments, the method also includes administering a tryptophan metabolism and / or kynurenine pathway modulator administered in (b) subsequent to initiation of administration of the T cell therapy.

[0033] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered at the following time points: when there is a decrease in tryptophan levels, an increase in kynurenine levels, and / or an increase in IDO1, IDO2, or TDO expression or activity in a biological sample from the subject compared to before administration of T cell therapy or compared to an earlier time point after administration of T cell therapy; before the number of T cell therapy cells detectable in the blood from the subject is decreased compared to in the subject at an earlier time point after administration of T cell therapy is initiated; before T cells exhibit prolonged tryptophan starvation, depletion, decreased effector function, expression of inhibitory receptors, and / or loss of proliferative capacity; before peak or maximum levels of T cell therapy cells are detectable in the subject's blood; and / or when levels of interferon-gamma (IFNγ) are increased in a biological sample from the subject compared to before administration of T cell therapy or compared to an earlier time point after administration of T cell therapy is initiated. In some embodiments, the increase or decrease is greater than 1.2-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold or more, or greater than about 1.2-fold, about 1.5-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 10-fold or more.

[0034] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator administered in (b) is administered within, or about, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 5 days, 6 days, 7 days, 14 days, 21 days, 28 days after initiation of T cell therapy. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator administered in (b) is administered within, or about, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, or 48 hours after initiation of T cell therapy.

[0035] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered at or about 0-96 hours, 0-72 hours, 0-48 hours, 0-24 hours, 0-12 hours, or 0-6 hours, or 0-2 hours prior to the initiation of T cell therapy, or the tryptophan metabolism and / or kynurenine pathway modulator is administered within 96 hours, 72 hours, 48 ​​hours, 24 hours, 12 hours, 6 hours, 2 hours, or 1 hour prior to the initiation of T cell therapy.

[0036] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is 1-methyl-D-tryptophan (1-MT) (indoximod), 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat ...}amino)-N-(3-chloro-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[4-fluoro-3-(trifluoromethyl)phenyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N'-hydroxy-N-[3-(trifluoromethyl)phenyl]- )phenyl]-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-cyano-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[(4-bromo-2-furyl)methyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4- ({2-[(aminosulfonyl)amino]ethyl}amino)-N-[(4-chloro-2-furyl)methyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 1-cyclohexyl-2-(5H-imidazo[5,1-a]isoindol-5-yl)ethanol (NLG919), GDC-0919, F001287, PF-06840003, or a pharmaceutically acceptable salt or prodrug thereof, or a combination thereof.

[0037] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is 1-methyl-D-tryptophan (1-MT) (indoximod). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is tryptophan.

[0038] Provided herein are methods of treatment that include: (a) administering a T cell therapy to a subject having a disease or condition; and (b) administering to the subject a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator, wherein the tryptophan metabolism and / or kynurenine pathway modulator is or includes 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat).

[0039] Also provided herein are methods of treatment comprising administering a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator to a subject having cancer who has been administered T cell therapy, wherein the tryptophan metabolism and / or kynurenine pathway modulator is or comprises 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat).

[0040] Also provided herein are methods of treatment comprising administering T cell therapy to a subject having cancer who has previously received a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator, wherein the tryptophan metabolism and / or kynurenine pathway modulator is or comprises 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat).

[0041] In some embodiments, the disease or condition is cancer.

[0042] In some embodiments of any of the embodiments herein, upon administration to a subject, the T cell therapy causes an increase or elevation in the level of IFNγ in the subject in the local environment of the tumor or in a serum or plasma sample of the subject compared to the level of IFNγ in the subject before initiation of T cell therapy.

[0043] In some embodiments of any of the embodiments herein, the T cell therapy is or comprises tumor infiltrating lymphocyte (TIL) therapy or T cell therapy comprising genetically engineered cells expressing a recombinant receptor that specifically binds to a ligand. In some embodiments, the T cell therapy is or comprises genetically engineered cells expressing a recombinant receptor that specifically binds to a ligand.

[0044] In some embodiments of any of the embodiments herein, the engineered cells also include modifications that affect the expression, presence, or activity of proteins or other factors or molecules associated with one or more activities, outcomes, or effects, and / or nucleic acids encoding such proteins or factors. In some aspects, the activities, outcomes, or effects may include those associated with, involved in sensing, or promoted by tryptophan deficiency or tryptophan starvation. In some aspects, the activities, outcomes, or effects may include those associated with, involved in sensing, or promoted by an increase in the amount or concentration of kynurenine or one or more kynurenine pathway metabolites and / or kynurenine-mediated immunosuppression, and / or a decrease in the tryptophan / kynurenine ratio in a cell or environment. In some embodiments, the protein or factor is or includes a protein involved in or associated with sensing tryptophan starvation and / or kynurenine-mediated immunosuppressive effects, such as those that may limit T cell function in a tumor environment. Such effects may include inhibiting or suppressing T cell activity or function, and / or promoting Treg function, levels, or activity. In some embodiments, the protein or factor is involved in IDO-induced immunosuppressive effects in cells and / or neighboring cells.

[0045] Provided herein are therapeutic methods comprising administering to a subject having a disease or condition genetically engineered T cells, wherein the genetically engineered T cells comprise (i) a recombinant receptor that specifically binds to a ligand, and (ii) modified expression of a protein involved in or associated with sensing tryptophan starvation and / or kynurenine-mediated immunosuppression in the cell, and / or a protein associated with IDO-mediated immunosuppressive effects.

[0046] Also provided herein are therapeutic methods comprising administering to a subject having a disease or condition genetically engineered T cells, wherein the genetically engineered T cells comprise (i) a recombinant receptor that specifically binds to a ligand and (ii) a recombinant, engineered, and / or ectopically expressed amino acid transporter or chain thereof, or a functional and / or catalytically active portion or variant thereof. In some embodiments, the method also comprises (b) administering to the subject a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator.

[0047] Provided herein are methods of treatment that include: (a) administering to a subject having a disease or condition genetically engineered T cells that comprise (i) a recombinant receptor that specifically binds to a ligand, and (ii) modified expression of a protein involved in or associated with sensing tryptophan starvation and / or kynurenine-mediated immunosuppressive effects in the cell, and / or a protein associated with IDO-mediated immunosuppressive effects; and (b) administering to the subject a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator.

[0048] In some embodiments, the disease or condition is cancer.

[0049] In some embodiments, the subject is selected as having a tumor positive for expression of L-type amino acid transporter 1 (LAT1; SLC7A5), LAT2 (SLC7A8), CD98 heavy chain (4F2hc; CD98hc; SLC3A2), and / or proton-assisted amino acid transporter 4 (PAT4; SLC36A4), or having cells in the tumor microenvironment positive for expression of LAT1, LAT2, CD98hc, and / or PAT4.

[0050] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator prevents or reduces the formation, concentration, availability, metabolism, and / or action of kynurenine pathway enzymes and / or metabolites. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator prevents or reduces the catabolism of tryptophan, or prevents or reduces the synthesis or accumulation of tryptophan metabolites. In some embodiments, the tryptophan metabolite is selected from kynurenine, 3-hydroxykynurenine, and 3-hydroxyanthranilic acid (3-HAA).

[0051] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is a small molecule, a peptide, a protein, an antibody or antigen-binding fragment thereof, an antibody mimetic, an aptamer, or a nucleic acid molecule.

[0052] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is an antagonist of the enzyme kynureninase.

[0053] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of one or more enzymes selected from the group consisting of inhibitors of indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1), IDO2, and inhibitors of tryptophan 2,3-dioxygenase (TDO). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a dual-acting IDO / TDO inhibitor.

[0054] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1). ...}amino)-N-(3-chloro-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[4-fluoro-3-(trifluoromethyl)phenyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N'-hydroxy-N-[3-(trifluoromethyl)phenyl]- )phenyl]-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-cyano-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[(4-bromo-2-furyl)methyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4- ({2-[(aminosulfonyl)amino]ethyl}amino)-N-[(4-chloro-2-furyl)methyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 1-cyclohexyl-2-(5H-imidazo[5,1-a]isoindol-5-yl)ethanol (NLG919), GDC-0919, F001287, PF-06840003, or a pharmaceutically acceptable salt or prodrug thereof, or a combination thereof.

[0055] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is 1-methyl-D-tryptophan (1-MT) (indoximod). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is tryptophan.

[0056] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is administered after initiation of administration of T cell therapy.

[0057] In some embodiments of any of the embodiments herein, the molecule, protein, or factor associated with IDO-mediated immunosuppressive signaling, associated with sensing or responding to tryptophan starvation or deficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression is mTOR or protein kinase C theta (PKC-θ).

[0058] In some embodiments, prior to initiation of administration of the composition comprising the T cells, the subject has failed or incompletely responded to treatment with prior administration of a tryptophan metabolism and / or kynurenine pathway modulator, has relapsed after remission following treatment, or has become refractory to treatment, where such prior administration was optionally in combination with an immunomodulatory agent, optionally in combination with a checkpoint inhibitor.

[0059] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered prior to, concurrently with, and / or after initiation of administration of T cell therapy, including administering the tryptophan metabolism and / or kynurenine pathway modulator after initiation of administration of T cell therapy.

[0060] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered at the following time points: when there is a decrease in tryptophan levels, an increase in kynurenine levels, and / or an increase in IDO1, IDO2, or TDO expression or activity in a biological sample from the subject compared to before administration of T cell therapy or compared to an earlier time point after administration of T cell therapy; before the number of T cell therapy cells detectable in the blood from the subject is decreased compared to in the subject at an earlier time point after administration of T cell therapy is initiated; before T cells exhibit prolonged tryptophan starvation, depletion, decreased effector function, expression of inhibitory receptors, and / or loss of proliferative capacity; before peak or maximum levels of T cell therapy cells are detectable in the subject's blood; and / or when levels of interferon-gamma (IFNγ) are increased in a biological sample from the subject compared to before administration of T cell therapy or compared to an earlier time point after administration of T cell therapy is initiated. In some embodiments, the increase or decrease is greater than 1.2-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold or more, or greater than about 1.2-fold, about 1.5-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 10-fold or more.

[0061] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered within, or about, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 5 days, 6 days, 7 days, 14 days, 21 days, 28 days after initiation of T cell therapy. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered within, or about, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, or 48 hours after initiation of T cell therapy.

[0062] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered at or about 0-96 hours, 0-72 hours, 0-48 hours, 0-24 hours, 0-12 hours, or 0-6 hours, or 0-2 hours prior to the initiation of T cell therapy, or the tryptophan metabolism and / or kynurenine pathway modulator is administered within 96 hours, 72 hours, 48 ​​hours, 24 hours, 12 hours, 6 hours, 2 hours, or 1 hour prior to the initiation of T cell therapy.

[0063] In some embodiments, the modification comprises recombinant, engineered, and / or ectopic expression of the molecule or a functionally and / or catalytically active chain, portion, or variant thereof. In some embodiments, the molecule is mTOR or protein kinase C theta (PKC-θ).

[0064] In some embodiments, the molecule is an amino acid transporter or a chain thereof, or a functional and / or catalytically active portion or variant thereof. In some embodiments, the amino acid transporter is a tryptophan transporter. In some embodiments, the amino acid transporter or a chain thereof is selected from one or more of rBAT (SLC3A1), CD98 heavy chain (4F2hc; SLC3A2), L-type amino acid transporter 1 (LAT1; SLC7A5), LAT2 (SLC7A8), Asc-type amino acid transporter 1 (Asc-1; SLC7A10), sodium- and chloride-dependent neutral and basic amino acid transporter B(0+) (ATB0,+; SLC6A14), sodium-dependent neutral amino acid transporter B(0)AT1 (B(0)AT1; SLC6A19), monocarboxylate transporter 10 (TAT1; SLC16A10), and proton-assisted amino acid transporter 4 (PAT4; SLC36A4) or portions thereof. In some embodiments, the amino acid transporter or chain thereof comprises CD98 heavy chain (4F2hc; SLC3A2) and L-type amino acid transporter 1 (LAT1; SLC7A5) or a portion thereof.

[0065] In some embodiments, the amino acid transporter or a chain thereof comprises an amino acid sequence selected from any one of SEQ ID NOs: 36-44 or a portion thereof, or an amino acid sequence or a portion thereof having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 36-44. In some embodiments, the amino acid transporter or a chain thereof is selected from among one or more, or portions thereof, of CD98 heavy chain (4F2hc; SLC3A2), L-type amino acid transporter 1 (LAT1; SLC7A5), LAT2 (SLC7A8), and proton-assisted amino acid transporter 4 (PAT4; SLC36A4). In some embodiments, the amino acid transporter or a chain thereof comprises CD98 heavy chain (4F2hc; SLC3A2) and L-type amino acid transporter 1 (LAT1; SLC7A5) or portions thereof.

[0066] In some embodiments, the amino acid transporter or a chain thereof comprises an amino acid sequence selected from any one of SEQ ID NOs: 37-39 and 44 or a portion thereof, or an amino acid sequence or a portion thereof having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 37-39 and 44. In some embodiments, expression of the molecule in the cell is under the control of a conditional promoter or enhancer or transactivator.

[0067] In some embodiments of any of the embodiments herein, the modification comprises recombinant, engineered, and / or ectopic expression of the protein or a functional and / or catalytically active portion or variant thereof. In some embodiments, expression of the protein in the cell is under the control of a conditional promoter or enhancer or transactivator.

[0068] In some embodiments of any of the embodiments herein, the molecule, protein, or factor involved in IDO-mediated immunosuppressive signal transduction, involved in sensing or responding to tryptophan starvation or deficiency, and / or involved in sensing or responding to kynurenine-mediated immunosuppression is GCN2 kinase, BLIMP-1, aryl hydrocarbon receptor (AHR), or AHR nuclear translocator (ARNT). In some embodiments, the molecule is selected from GCN2 kinase, BLIMP-1, aryl hydrocarbon receptor (AHR), AHR nuclear translocator (ARNT), eukaryotic translation initiation factor 2α (eIF2α), activating transcription factor 4 (ATF4), or CCAAT / enhancer-binding protein homologous protein (CHOP), Gadd45α, Herp, 4E-BP1, eIF4G, JAK1, NFκB-2, FK506-BP8, and IFNγ-R2. In some embodiments, the molecule is GCN2 or CHOP.

[0069] In some embodiments of any of the embodiments herein, the modification comprises reduced expression of a molecule. In some embodiments, the engineered cell comprises an inhibitory nucleic acid or gene disruption that reduces expression of the molecule. In some embodiments, the expression of the molecule in the cell is reduced by at least 50, 60, 70, 80, 90, or 95% compared to expression in the cell in the absence of the agent or gene disruption.

[0070] In some embodiments of any of the methods provided herein, the method comprises introducing an inhibitory nucleic acid into a cell, thereby resulting in reduced expression of the molecule. In some embodiments, the engineered cell comprises an inhibitory nucleic acid, and the inhibitory nucleic acid comprises an RNA interfering agent. In some embodiments, the inhibitory nucleic acid is, comprises, or encodes a small interfering RNA (siRNA), a microRNA-matched shRNA, a short hairpin RNA (shRNA), a hairpin siRNA, a microRNA (miRNA precursor), or a microRNA (miRNA). In some embodiments, the method comprises introducing a nucleic acid that encodes or produces the inhibitory nucleic acid into a cell, thereby resulting in reduced expression of the molecule.

[0071] In some embodiments of any of the embodiments herein, expression of the inhibitory nucleic acid is under the control of a conditional promoter or enhancer or transactivator.

[0072] In some embodiments of any of the embodiments herein, the engineered cell comprises a gene disruption, wherein the disruption comprises disrupting the gene encoding the molecule at the DNA level, and / or the disruption is not reversible, and / or the disruption is not transient. In some embodiments, the disruption comprises introducing into the cell a DNA-binding protein or a DNA-binding nucleic acid that specifically binds to or hybridizes to the gene encoding the molecule.

[0073] In some embodiments of any of the embodiments herein, the disruption comprises introducing (a) a fusion protein comprising a DNA-targeting protein and a nuclease, or (b) an RNA-guided nuclease. In some embodiments, the DNA-targeting protein or RNA-guided nuclease comprises a zinc finger protein (ZFP), a TAL protein, or a CRISPR (clustered regularly interspaced short palindromic nucleic acid) specific to the gene. In some embodiments, the disruption comprises introducing into the cell an agent comprising a zinc finger nuclease (ZFN), a TAL effector nuclease (TALEN), or a CRISPR-Cas9 combination that specifically binds to, recognizes, or hybridizes with the gene encoding the molecule. In some embodiments, the agent is a CRISPR-Cas9 combination, and the CRISPR-Cas9 combination comprises a guide RNA (gRNA) comprising a guide sequence that is complementary to and / or can hybridize with a target sequence within the gene.

[0074] In some embodiments of any of the embodiments herein, the genetic disruption is inducible. In some embodiments, the CRISPR-Cas9 complex is an inducible CRISPR-Cas9, and / or the Cas9 is under the control of a conditional promoter, enhancer, or transactivator.

[0075] In some embodiments of any of the embodiments herein, the disruption comprises a deletion of at least a portion of at least one exon of the gene encoding the molecule.

[0076] In some embodiments of any of the embodiments herein, the disruption comprises a deletion, mutation, and / or insertion within the gene that results in the presence of a premature stop codon within the gene, and / or the disruption comprises a deletion, mutation, and / or insertion within the first or second exon of the gene encoding the molecule.

[0077] In some embodiments of any of the embodiments herein, the conditional promoter, enhancer, or transactivator is an inducible promoter, enhancer, or transactivator, or a repressible promoter, enhancer, or transactivator. In some embodiments, the promoter is selected from among RNA pol I, pol II, or pol II promoters. In some embodiments, the promoter is selected from a pol III promoter that is a U6 or H1 promoter, or a pol II promoter that is a CMV, SV40 early region, or adenovirus major late promoter.

[0078] In some embodiments of any of the embodiments herein, the promoter is an inducible promoter, hi some embodiments, the promoter comprises a Lac operator sequence, a tetracycline operator sequence, a galactose operator sequence, or a doxycycline operator sequence, or an analog thereof.

[0079] In some embodiments of any of the embodiments herein, the promoter is a repressible promoter, ie, the promoter can be bound by a Lac repressor or a tetracycline repressor, or an analog thereof.

[0080] In some embodiments of any of the embodiments herein, the engineered receptor that specifically binds to a ligand is a functional non-T cell receptor. In some embodiments, the engineered receptor that specifically binds to a ligand is a chimeric antigen receptor (CAR). In some embodiments, the CAR comprises an extracellular antigen recognition domain that specifically binds to an antigen and an intracellular signaling region comprising an ITAM. In some embodiments, the intracellular signaling region comprises the intracellular domain of the CD3-zeta (CD3ζ) chain.

[0081] In some embodiments of any of the embodiments herein, the CAR also comprises a costimulatory signaling region. In some embodiments, the costimulatory signaling region comprises the signaling domain of CD28 or 4-1BB or ICOS, or a signaling portion thereof. In some embodiments, the costimulatory signaling region is the signaling domain of CD28.

[0082] In some embodiments of any of the embodiments herein, the recombinant receptor is a transgenic T cell receptor (TCR). In some embodiments, the recombinant receptor is a chimeric antigen receptor (CAR).

[0083] In some embodiments of any of the embodiments herein, the T cell therapy recognizes or targets an antigen associated with cancer. In some embodiments, the recombinant receptor binds to, recognizes, or targets an antigen associated with cancer. In some embodiments, the antigen is selected from ROR1, Her2, L1-CAM, CD19, CD20, CD22, mesothelin, CEA, hepatitis B surface antigen, antifolate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, EGP-2, EGP-4, EPHa2, ErbB2, ErbB3, ErbB4, FBP, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-α, IL-1 Selected from 3R-α2, kdr, kappa light chain, Lewis Y, L1 cell adhesion molecule, MAGE-A1, mesothelin, MUC1, MUC16, PSCA, NKG2D ligand, NY-ESO-1, MART-1, gp100, oncofetal antigen, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), prostate-specific antigen, PSMA, estrogen receptor, progesterone receptor, ephrin B2, CD123, CS-1, c-Met, GD-2, MAGE A3, CE7, Wilms' tumor 1 (WT-1), and cyclin A1 (CCNA1).

[0084] In some embodiments of any of the embodiments herein, the cancer comprises a tumor negative for indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1), and / or the subject has not been selected for the development of an IDO1-positive tumor. In some embodiments, the cancer comprises a tumor negative for indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1) prior to initiation of administration of T cell therapy, and / or the subject has not been selected for the development of an IDO1-positive tumor prior to initiation of administration of T cell therapy. In some embodiments, the cancer comprises a tumor positive for expression of L-type amino acid transporter 1 (LAT1; SLC7A5), LAT2 (SLC7A8), CD98 heavy chain (4F2hc; CD98hc; SLC3A2), and / or proton-assisted amino acid transporter 4 (PAT4; SLC36A4), or the cancer comprises cells in the tumor microenvironment positive for expression of LAT1, LAT2, CD98hc, and / or PAT4.

[0085] In some embodiments of any of the embodiments herein, the cancer is myeloma, lymphoma, or leukemia. In some embodiments, the cancer is non-Hodgkin's lymphoma (NHL), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), or myeloma. In some embodiments, the cancer does not express B-cell antigens or is not a B-cell malignancy. In some embodiments, the cancer does not express CD19, and / or the T-cell therapy does not comprise a recombinant receptor that specifically binds to CD19, and / or the T-cell therapy is a CAR-T cell therapy that does not comprise an anti-CD19 antigen-binding domain.

[0086] In some embodiments of any of the aspects herein, the cancer is a non-hematological cancer or a solid tumor.

[0087] In some embodiments, the methods of the invention result in increased tryptophan levels and / or decreased kynurenine levels in a tumor or in a biological sample from a subject, optionally in serum, plasma, or tumor sample, compared to methods involving administration of T cell therapy but in the absence of a tryptophan metabolism and / or kynurenine pathway modulator, or compared to methods in which cells are administered that do not include altered expression of molecules associated with IDO-mediated immunosuppressive signaling, associated with sensing or responding to tryptophan starvation or deficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression.

[0088] In some embodiments of any of the embodiments herein, the T cell therapy exhibits increased or long-term expansion and / or persistence in the subject compared to methods in which the T cell therapy is administered to the subject in the absence of the tryptophan metabolism and / or kynurenine pathway modulator.

[0089] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is administered orally, subcutaneously, or intravenously. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered orally.

[0090] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator may be present in an amount of 2.5 mg to 5000 mg, 2.5 mg to 2000 mg, 2.5 mg to 1000 mg, 2.5 mg to 500 mg, 2.5 mg to 200 mg, 2.5 mg to 100 mg, 2.5 mg to 50 mg, 2.5 mg to 25 mg, 2.5 mg to 10 mg, 25 mg to 5000 mg, 25 mg to 2000 mg, 25 mg to 1000 mg, 25 mg to 500 mg, 25 mg to 200 mg, 25 mg to 100 mg, 25 mg to 50 mg, 50 mg, inclusive. to 5000mg, 50mg to 2000mg, 50mg to 1000mg, 50mg to 500mg, 50mg to 200mg, 50mg to 100mg, 100mg to 5000mg, 100mg to 2000mg, 100mg to 1000mg, 100mg to 500mg, 100mg to 200mg, 200mg to 5000mg, 200mg to 2000mg, 200mg to 1000mg, 200mg to 500mg, 500mg to 5000mg, 500mg to 2000mg, 500mg to 1000mg, or 1000mg to 2000mg. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered six times per day, five times per day, four times per day, three times per day, twice per day, or once per day. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered at a total daily dosage of at least or at least about 5 mg / day, 10 mg / day, 25 mg / day, 50 mg / day, 100 mg / day, 200 mg / day, 400 mg / day, 500 mg / day, 600 mg / day, 800 mg / day, 1000 mg / day, 1200 mg / day, 1600 mg / day, 2000 mg / day, 5000 mg / day, or 10000 mg / day.

[0091] In some embodiments of any of the embodiments herein, administration of the tryptophan metabolism and / or kynurenine pathway modulator is continued until, after initiation of administration of the T cell therapy, there is: an increase in tryptophan levels, a decrease in kynurenine levels, and / or a decrease in IDO1, IDO2, or TDO expression or activity in a biological sample from the subject compared to immediately before administration of the tryptophan metabolism and / or kynurenine pathway modulator or compared to immediately before initiation of administration of the T cell therapy; The administration of T cell therapy is continued until the number of T cell therapy cells detectable in the blood is increased compared to that in the subject at a prior time point immediately prior to administration of the T cell pathway modulator or compared to a prior time point after administration of the T cell therapy; until the number of T cell therapy cells detectable in the blood is within (approximately) 2.0 times the peak or maximum number of T cell therapy cells detectable in the subject's blood after initiation of administration of the T cell therapy; and / or until the number of T cell therapy cells detectable in the blood from the subject exceeds, or exceeds approximately, 10%, 15%, 20%, 30%, 40%, 50%, or 60% of the total peripheral blood mononuclear cells (PBMCs) in the subject's blood.

[0092] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is administered until the concentration or number of engineered cells in the subject's blood is reduced to: (i) at least 10 or at least about 10 engineered cells per microliter; (ii) at least 20%, 30%, 40%, or 50% of the total number of peripheral blood mononuclear cells (PBMCs); (iii) at least 1 x 10 5 or at least about 1×10 5engineered cells, or (iv) until the blood contains at least 5,000 copies of recombinant receptor-encoding DNA per microgram of DNA; and / or until 90 days after initiation of administration of (a), CAR-expressing cells are detectable in the subject's blood or serum; and / or until 90 days after initiation of administration of (a), the subject's blood contains at least 20% CAR-expressing cells, at least 10 CAR-expressing cells per microliter, or at least 1 x 10 4 The cells are administered until they contain 100 CAR-expressing cells.

[0093] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is administered for a period of up to 2 days, up to 7 days, up to 14 days, up to 21 days, up to 1 month, up to 2 months, up to 3 months, up to 6 months, or up to 1 year after initiation of administration of the T cell therapy.

[0094] In some embodiments of any of the embodiments herein, the T cell therapy comprises T cells that are CD4+ or CD8+. In some embodiments, the T cell therapy comprises cells that are autologous to the subject. In some embodiments, the T cell therapy comprises T cells that are allogeneic to the subject.

[0095] In some embodiments of any of the embodiments herein, the T cell therapy is administered in an amount that upregulates IDO1 expression in the tumor microenvironment. In some embodiments, IDO1 expression is upregulated in or from bone marrow cells, stromal cells, or tumor cells. In some embodiments, IDO1 expression is upregulated in or from bone marrow stromal cells.

[0096] In some embodiments of any of the methods provided herein, the cell therapy is administered at a dose of 0.5 x 10 per kg of the subject's body weight, inclusive. 6 cells / kg~5×10 6 cells / kg or approximately 0.5 x 10 6 cells / kg ~ approx. 5×10 6 Cells / kg, 0.5 x 10 cells / kg of subject body weight6 cells / kg~3×10 6 cells / kg or approximately 0.5 x 10 6 cells / kg ~ approx. 3×10 6 Cells / kg, 0.5 x 10 cells / kg of subject body weight 6 cells / kg~2×10 6 cells / kg or approximately 0.5 x 10 6 cells / kg ~ approx. 2×10 6 Cells / kg, 0.5 x 10 cells / kg of subject body weight 6 cells / kg~1×10 6 cells / kg or approximately 0.5 x 10 6 cells / kg ~ approx. 1×10 6 Cells / kg, 1.0 x 10 per kg of subject body weight 6 cells / kg~5×10 6 cells / kg or approximately 1.0 x 10 6 cells / kg ~ approx. 5×10 6 Cells / kg, 1.0 x 10 per kg of subject body weight 6 cells / kg~3×10 6 cells / kg or approximately 1.0 x 10 6 cells / kg ~ approx. 3×10 6 Cells / kg, 1.0 x 10 per kg of subject body weight 6 cells / kg~2×10 6 cells / kg or approximately 1.0 x 10 6 cells / kg ~ approx. 2×10 6 Cells / kg, 2.0 x 10 per kg of subject body weight 6 cells / kg~5×10 6 cells / kg or approximately 2.0 x 10 6 cells / kg ~ approx. 5×10 6 Cells / kg, 2.0 x 10 per kg of subject body weight 6 cells / kg~3×10 6 cells / kg or approximately 2.0 x 10 6 cells / kg ~ approx. 3×10 6 Cells / kg, 3.0 × 10 cells / kg of subject body weight 6 cells / kg~5×10 6 cells / kg or approximately 3.0 x 10 6 cells / kg ~ approx. 5×10 6This involves administering a dose containing a cell number of cells / kg.

[0097] In some embodiments of any of the embodiments herein, the dose of cells administered is less than the dose in methods in which the cell therapy is administered without administering a tryptophan metabolism and / or kynurenine pathway modulator, or in methods in which the cells do not include altered expression of molecules associated with IDO-mediated immunosuppressive signaling, associated with sensing or responding to tryptophan starvation or deficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression. In some embodiments, the dose is at least 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, or 10-fold less.

[0098] In some embodiments of any of the methods provided herein, the T cell therapy is administered as a single pharmaceutical composition comprising the cells. In some embodiments, the T cell therapy comprises cell doses that are split doses, wherein the doses of cells are administered in multiple compositions collectively comprising the doses of cells over a period of no more than 3 days.

[0099] In some embodiments of any of the aspects herein, the method further comprises administering lymphodepleting chemotherapy prior to administering T cell therapy. In some embodiments, the method does not comprise administering fludarabine to the subject. In some embodiments of any of the aspects herein, the provided method does not comprise administering lymphodepleting chemotherapy to the subject prior to administering T cell therapy.

[0100] In some embodiments, the recombinant receptor has been engineered by introducing into a cell a first nucleic acid sequence encoding the recombinant receptor and one or more second nucleic acid sequences encoding agents capable of or involved in modifying the expression of molecules associated with IDO-mediated immunosuppressive signaling in the cell, associated with sensing or responding to tryptophan starvation or deficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression. In some embodiments, the first nucleic acid sequence and the second nucleic acid sequence(s) are comprised in one or more polynucleotides. In some embodiments, the first nucleic acid sequence and the second nucleic acid sequence(s) are comprised in two polynucleotides. In some embodiments, the first nucleic acid and the second nucleic acid(s) are comprised in one or more vectors, optionally viral vector(s).

[0101] Provided herein are methods of selecting a subject having a disease or condition for administration of a tryptophan metabolism and / or kynurenine pathway modulator, the methods comprising: (a) assessing the level of tryptophan or a tryptophan metabolite, the level of expression or activity of IDO1, IDO2, or TDO, or the level of interferon-gamma (IFNγ) in one or more biological samples from a subject who is a candidate for treatment with T cell therapy; and (b) selecting subjects in which: (i) the level of tryptophan in the sample is below a threshold level; (ii) the level of a tryptophan metabolite is above a threshold level; (iii) the level of expression or activity of IDO1, IDO2, or TDO is above a threshold level; or (iv) the level of IFNγ is above a threshold level.

[0102] In some embodiments, the one or more biological samples are obtained prior to administration of the T cell therapy.

[0103] In some embodiments, the method also includes administering T cell therapy to the subject. In some embodiments, the method also includes administering a tryptophan metabolism and / or kynurenine pathway modulator to the selected subject. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered to the selected subject before or simultaneously with the initiation of administration of T cell therapy. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered after the initiation of administration of T cell therapy.

[0104] Provided herein are methods for selecting a subject having a disease or condition for administration of a tryptophan metabolism and / or kynurenine pathway modulator, the methods comprising: (a) assessing the level of tryptophan or a tryptophan metabolite, the level of IDO1, IDO2, or TDO expression or activity, or the level of interferon-gamma (IFNγ) in one or more biological samples obtained from the subject who has been administered T cell therapy; and (b) determining whether (i) the level of tryptophan in the sample is below a threshold level or is decreased compared to the level assessed at a time point before or at an earlier time point after initiation of administration of T cell therapy. (ii) the level of a tryptophan metabolite is above a threshold level or is increased compared to the level assessed before or at an earlier time point after initiation of administration of T cell therapy; (iii) the level of IDO1, IDO2, or TDO expression or activity is above a threshold level or is increased compared to the level assessed before or at an earlier time point after initiation of administration of T cell therapy; or (iv) the level of IFNγ is above a threshold level or is increased compared to the level assessed before or at an earlier time point after initiation of administration of T cell therapy.

[0105] In some embodiments, the one or more biological samples are obtained after administration of T cell therapy. In some embodiments, the method also includes administering a cell therapy to the subject prior to the assessment. In some embodiments, the method also includes administering a kynurenine pathway modulator to the subject.

[0106] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered at the following time points: when there is a decrease in tryptophan levels, an increase in kynurenine levels, and / or an increase in IDO1, IDO2, or TDO expression or activity in a biological sample from the subject compared to before administration of T cell therapy or compared to an earlier time point after administration of T cell therapy; before the number of T cell therapy cells detectable in the blood from the subject is decreased compared to in the subject at an earlier time point after administration of T cell therapy is initiated; before T cells exhibit prolonged tryptophan starvation, depletion, decreased effector function, expression of inhibitory receptors, and / or loss of proliferative capacity; before peak or maximum levels of T cell therapy cells are detectable in the subject's blood; and / or when levels of interferon-gamma (IFNγ) are increased in a biological sample from the subject compared to before administration of T cell therapy or compared to an earlier time point after administration of T cell therapy is initiated. In some embodiments, the increase or decrease is greater than 1.2-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold or more, or greater than about 1.2-fold, about 1.5-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 10-fold or more.

[0107] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered within, or about, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 5 days, 6 days, 7 days, 14 days, 21 days, 28 days after initiation of T cell therapy. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered within, or about, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, or 48 hours after initiation of T cell therapy.

[0108] In some embodiments, the disease or condition is cancer.

[0109] In some embodiments, the subject is selected if the level of expression or activity of IDO1, IDO2, or TDO is increased compared to the level assessed at a time point before administration of T cell therapy began. In some embodiments, the subject is selected if the level of tryptophan in the sample is below a threshold level or is decreased compared to the level assessed at a time point before administration of T cell therapy began.

[0110] In some embodiments, the threshold level is within 25%, 20%, 15%, 10%, or 5% of the mean level, concentration, or amount in biological samples from multiple control subjects, and / or within the standard deviation of the mean level, concentration, or amount. In some embodiments, the control subjects are healthy or normal subjects, subjects without cancer, and / or subjects prior to receiving T cell therapy.

[0111] In some embodiments, the increase or decrease is greater than 1.2-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold or more, or greater than about 1.2-fold, about 1.5-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 10-fold or more.

[0112] In some embodiments, prior to assessment, the subject has failed to respond or has not fully responded to treatment with prior administration of a tryptophan metabolism and / or kynurenine pathway modulator, has relapsed after remission following treatment, or has become refractory to treatment, where such prior administration was optionally in combination with an immunomodulatory agent, optionally in combination with a checkpoint inhibitor.

[0113] In some embodiments, the biological sample is or is derived from serum, plasma, or a tumor sample.

[0114] In some embodiments, the tryptophan metabolite is selected from kynurenine, 3-hydroxykynurenine, and 3-hydroxyanthranilic acid (3-HAA).

[0115] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is a small molecule, a peptide, a protein, an antibody or antigen-binding fragment thereof, an antibody mimetic, an aptamer, or a nucleic acid molecule.

[0116] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is an antagonist of the enzyme kynureninase.

[0117] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of one or more enzymes selected from the group consisting of inhibitors of indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1), IDO2, and inhibitors of tryptophan 2,3-dioxygenase (TDO). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a dual-acting IDO / TDO inhibitor.

[0118] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1). ...}amino)-N-(3-chloro-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[4-fluoro-3-(trifluoromethyl)phenyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N'-hydroxy-N-[3-(trifluoromethyl)phenyl]- )phenyl]-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-cyano-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[(4-bromo-2-furyl)methyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4- ({2-[(aminosulfonyl)amino]ethyl}amino)-N-[(4-chloro-2-furyl)methyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 1-cyclohexyl-2-(5H-imidazo[5,1-a]isoindol-5-yl)ethanol (NLG919), GDC-0919, F001287, PF-06840003, or a pharmaceutically acceptable salt or prodrug thereof, or a combination thereof.

[0119] In some embodiments of any of the embodiments herein, the tryptophan metabolism and / or kynurenine pathway modulator is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is 1-methyl-D-tryptophan (1-MT) (indoximod). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is tryptophan.

[0120] In some embodiments, the T cells comprise a recombinant receptor that specifically binds to a ligand that is a functional non-T cell receptor.

[0121] In some embodiments of any of the embodiments herein, the engineered receptor that specifically binds to a ligand is a functional non-T cell receptor. In some embodiments, the engineered receptor that specifically binds to a ligand is a chimeric antigen receptor (CAR). In some embodiments, the CAR comprises an extracellular antigen recognition domain that specifically binds to an antigen and an intracellular signaling region comprising an ITAM. In some embodiments, the intracellular signaling region comprises the intracellular domain of the CD3-zeta (CD3ζ) chain.

[0122] In some embodiments of any of the embodiments herein, the CAR also comprises a costimulatory signaling region. In some embodiments, the costimulatory signaling region comprises the signaling domain of CD28 or 4-1BB or ICOS, or a signaling portion thereof. In some embodiments, the costimulatory signaling region is the signaling domain of CD28.

[0123] In some embodiments of any of the embodiments herein, the recombinant receptor is a transgenic T cell receptor (TCR). In some embodiments, the recombinant receptor is a chimeric antigen receptor (CAR).

[0124] In some embodiments of any of the embodiments herein, the T cell therapy recognizes or targets an antigen associated with cancer. In some embodiments, the recombinant receptor binds to, recognizes, or targets an antigen associated with cancer. In some embodiments, the antigen is selected from ROR1, Her2, L1-CAM, CD19, CD20, CD22, mesothelin, CEA, hepatitis B surface antigen, antifolate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, EGP-2, EGP-4, EPHa2, ErbB2, ErbB3, ErbB4, FBP, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-α, IL-1 Selected from 3R-α2, kdr, kappa light chain, Lewis Y, L1 cell adhesion molecule, MAGE-A1, mesothelin, MUC1, MUC16, PSCA, NKG2D ligand, NY-ESO-1, MART-1, gp100, oncofetal antigen, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), prostate-specific antigen, PSMA, estrogen receptor, progesterone receptor, ephrin B2, CD123, CS-1, c-Met, GD-2, MAGE A3, CE7, Wilms' tumor 1 (WT-1), and cyclin A1 (CCNA1).

[0125] Provided herein are engineered cells comprising: (a) a genetically engineered receptor that specifically binds to a ligand; and (b) modified expression of a molecule associated with IDO-mediated immunosuppressive signaling in the cell, associated with sensing or responding to tryptophan starvation or deficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression.

[0126] In some embodiments, the molecule is mTOR or protein kinase C theta (PKC-θ).

[0127] Engineered cells are provided that comprise: (a) a recombinant receptor that specifically binds to a ligand; and (b) modified expression of a molecule associated with IDO-mediated immunosuppressive signaling in the cell, associated with sensing or responding to tryptophan starvation or deficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression.

[0128] In some embodiments, the modification comprises recombinant, engineered, and / or ectopic expression in the cell of the molecule or a functionally and / or catalytically active chain, portion, or variant thereof. In some embodiments, the molecule is mTOR or protein kinase C theta (PKC-θ).

[0129] In some embodiments, the molecule is an amino acid transporter or a chain thereof, or a functional and / or catalytically active portion or variant thereof.

[0130] Also provided are engineered cells comprising: (a) a recombinant receptor that specifically binds to a ligand; and (b) a recombinant, engineered, and / or ectopically expressed amino acid transporter or chain thereof, or a functional and / or catalytically active portion or variant thereof.

[0131] In some embodiments, the amino acid transporter is a tryptophan transporter. In some embodiments, the amino acid transporter is selected from the group consisting of T (SLC3A1), CD98 heavy chain (4F2hc; SLC3A2), L-type amino acid transporter 1 (LAT1; SLC7A5), LAT2 (SLC7A8), Asc-type amino acid transporter 1 (Asc-1; SLC7A10), sodium- and chloride-dependent neutral and basic amino acid transporter B(0+) (ATB0,+; SLC6A14), sodium-dependent neutral amino acid transporter B(0)AT1 (B(0)AT1; SLC6A19), monocarboxylate transporter 10 (TAT1; SLC16A10), and proton-assisted amino acid transporter 4 (PAT4; SLC36A4), or portions thereof. In some embodiments, the amino acid transporter or chain thereof comprises CD98 heavy chain (4F2hc; SLC3A2) and L-type amino acid transporter 1 (LAT1; SLC7A5) or a portion thereof.

[0132] In some embodiments, the amino acid transporter or a chain thereof comprises an amino acid sequence selected from any one of SEQ ID NOs: 36-115, or an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 36-115. In some embodiments, the amino acid transporter or a chain thereof is selected from one or more of CD98 heavy chain (4F2hc; SLC3A2), L-type amino acid transporter 1 (LAT1; SLC7A5), LAT2 (SLC7A8), and proton-assisted amino acid transporter 4 (PAT4; SLC36A4), or portions thereof. In some embodiments, the amino acid transporter or a chain thereof comprises CD98 heavy chain (4F2hc; SLC3A2) and L-type amino acid transporter 1 (LAT1; SLC7A5), or portions thereof.

[0133] In some embodiments, the amino acid transporter or a chain thereof comprises an amino acid sequence selected from any one of SEQ ID NOs: 37-39 and 115, or an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 37-39 and 115.

[0134] In some embodiments, expression of the molecule or amino acid transporter in the cell is under the control of a conditional promoter or enhancer or transactivator.

[0135] In some embodiments, the molecule is selected from among GCN2 kinase, BLIMP-1, aryl hydrocarbon receptor (AHR), AHR nuclear translocator (ARNT), eukaryotic translation initiation factor 2 alpha (eIF2α), activating transcription factor 4 (ATF4), or CCAAT / enhancer-binding protein homologous protein (CHOP), Gadd45α, Herp, 4E-BP1, eIF4G, JAK1, NFκB-2, FK506-BP8, and IFNγ-R2. In some embodiments, the molecule is GCN2 or CHOP.

[0136] In some embodiments, the modification comprises recombinant, engineered, and / or ectopic expression in the cell of the molecule or a functionally and / or catalytically active chain, portion, or variant thereof, hi some embodiments, expression of the molecule in the cell is under the control of a conditional promoter or enhancer or transactivator.

[0137] In some embodiments, the molecule is GCN2 kinase, aryl hydrocarbon receptor (AHR), or AHR nuclear translocator (ARNT).

[0138] In some embodiments, the modification comprises reducing the expression of a molecule in the cell. In some embodiments, the engineered cell comprises an inhibitory nucleic acid or gene disruption that reduces the expression of the molecule. In some embodiments, the expression of the molecule in the cell is reduced by at least 50, 60, 70, 80, 90, or 95% compared to expression in the cell in the absence of the inhibitory nucleic acid or gene disruption.

[0139] In some embodiments, the cell contains an inhibitory nucleic acid, thereby reducing the expression of the molecule.In some embodiments, the inhibitory nucleic acid comprises an RNA interfering agent.In some embodiments, the inhibitory nucleic acid is, comprises, or encodes small interfering RNA (siRNA), microRNA-compatible shRNA, short hairpin RNA (shRNA), hairpin siRNA, microRNA (miRNA precursor) or microRNA (miRNA).In some embodiments, the expression of the inhibitory nucleic acid is under the control of a conditional promoter or enhancer or transactivator.

[0140] In some embodiments, the engineered cell comprises a disrupted gene encoding the molecule, an agent to disrupt the gene encoding the molecule, and / or disruption of the gene encoding the molecule. In some embodiments, the disruption comprises disrupting the gene encoding the molecule at the DNA level, and / or the disruption is not reversible, and / or the disruption is not transient.

[0141] In some embodiments, the disruption is mediated by a DNA-binding protein or DNA-binding nucleic acid that specifically binds to or hybridizes with the gene encoding the molecule. In some embodiments, the disruption is mediated by (a) a fusion protein comprising a DNA-targeting protein and a nuclease, or (b) an RNA-guided nuclease. In some embodiments, the disruption is mediated by a gene-editing nuclease, a zinc finger nuclease (ZFN), a TAL effector nuclease (TALEN), or a clustered regularly interspaced short palindromic nucleic acid (CRISPR) / Cas9. In some embodiments, the disruption is mediated by CRISPR / Cas9, and CRISPR / Cas9 comprises a guide RNA (gRNA) that comprises a guide sequence that is complementary to and / or can hybridize with a target sequence in the gene.

[0142] In some embodiments, the genetic disruption is conditional or inducible. In some embodiments, the CRISPR-Cas9 complex is an inducible CRISPR-Cas9 and / or the Cas9 is under the control of a conditional promoter or enhancer or transactivator.

[0143] In some embodiments, the disruption comprises a deletion of at least a portion of at least one exon of the gene encoding the molecule. In some embodiments, the disruption comprises a deletion, mutation, and / or insertion within the gene that results in the presence of a premature stop codon within the gene; and / or the disruption comprises a deletion, mutation, and / or insertion within the first or second exon of the gene encoding the molecule.

[0144] In some embodiments, the promoter is an inducible promoter, enhancer, or transactivator, such as an RNA pol I, pol II, or pol III promoter, or a repressible promoter, enhancer, or transactivator. In some embodiments, the promoter is selected from a pol III promoter that is a U6 or H1 promoter, or a pol II promoter that is a CMV, SV40 early region, or adenovirus major late promoter.

[0145] In some embodiments, the promoter is an inducible promoter. In some embodiments, the promoter contains binding sites for NFκB or NFAT. In some embodiments, the promoter contains a Lac operator sequence, a tetracycline operator sequence, a galactose operator sequence, or a doxycycline operator sequence, or an analog thereof.

[0146] In some embodiments, the promoter is a repressible promoter, hi some embodiments, the promoter can be bound by a Lac repressor or a tetracycline repressor, or an analog thereof.

[0147] In some embodiments, the cells are T cells. In some embodiments, the T cells are CD8+ T cells. In some embodiments, the T cells are CD4+ T cells. In some embodiments, the cells are natural killer (NK) cells. In some embodiments, the cells are iPS-derived cells.

[0148] In some embodiments, the engineered receptor that specifically binds to a ligand is a functional non-T cell receptor, hi some embodiments, the engineered receptor that specifically binds to a ligand is a chimeric antigen receptor (CAR).

[0149] In some embodiments, the CAR comprises an extracellular antigen recognition domain that specifically binds to an antigen and an intracellular signaling region that comprises an ITAM.

[0150] In some embodiments, the intracellular signaling region comprises the intracellular domain of the CD3-zeta (CD3ζ) chain.

[0151] In some embodiments, the CAR also comprises a costimulatory signaling region. In some embodiments, the costimulatory signaling region comprises the signaling domain of CD28 or 4-1BB or ICOS, or a signaling portion thereof. In some embodiments, the costimulatory signaling region is the signaling domain of CD28.

[0152] In some embodiments, the engineered receptor that specifically binds to a ligand is a recombinant receptor.

[0153] In some embodiments, the engineered receptor that specifically binds to a ligand is a transgenic T cell receptor (TCR).

[0154] In some embodiments, the cells have been engineered by introducing into the cells a first nucleic acid sequence encoding a recombinant receptor and one or more second nucleic acid sequences encoding agents capable of modifying or involved in the expression of molecules associated with IDO-mediated immunosuppressive signaling in the cells, associated with sensing or responding to tryptophan starvation or deficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression. In some embodiments, the first nucleic acid sequence and the second nucleic acid sequence(s) are comprised in one or more polynucleotides. In some embodiments, the first nucleic acid sequence and the second nucleic acid sequence(s) are comprised in two polynucleotides. In some embodiments, the first nucleic acid sequence and the second nucleic acid sequence(s) are comprised in one or more vectors, optionally viral vector(s).

[0155] Also provided herein are compositions comprising any of the engineered cells described herein. In some embodiments of the compositions, the cells are CD4+ or CD8+ cells. In some embodiments, the compositions also include a pharmaceutically acceptable carrier.

[0156] Also provided herein is a combination comprising a genetically engineered cell expressing a recombinant receptor that binds a ligand, optionally a T cell receptor (TCR) or a chimeric antigen receptor (CAR); and a tryptophan metabolism and / or kynurenine pathway modulator.

[0157] Also provided herein are combinations comprising any of the engineered cells described herein or any of the compositions described herein and a tryptophan metabolism and / or kynurenine pathway modulator.

[0158] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator prevents or reduces the formation, concentration, availability, metabolism, and / or action of kynurenine pathway enzymes and / or metabolites and / or reduces the kynurenine / tryptophan ratio.

[0159] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator prevents or reduces the catabolism of tryptophan or prevents or reduces the synthesis or accumulation of a tryptophan metabolite, hi some embodiments, the tryptophan metabolite is selected from kynurenine, 3-hydroxykynurenine, and 3-hydroxyanthranilic acid (3-HAA).

[0160] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a small molecule, peptide, protein, antibody or antigen-binding fragment thereof, antibody mimetic, aptamer, or nucleic acid molecule.

[0161] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an antagonist of the enzyme kynureninase.

[0162] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of one or more enzymes selected from the group consisting of inhibitors of indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1), IDO2, and inhibitors of tryptophan 2,3-dioxygenase (TDO). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a dual-acting IDO / TDO inhibitor. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1).

[0163] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is 1-methyl-D-tryptophan (1-MT) (indoximod), 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N′-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat), 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-( 3-chloro-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[4-fluoro-3-(trifluoromethyl)phenyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N'-hydroxy-N-[3-(trifluoromethyl)phenyl] -1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-cyano-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[(4-bromo-2-furyl)methyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2 -[(aminosulfonyl)amino]ethyl}amino)-N-[(4-chloro-2-furyl)methyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 1-cyclohexyl-2-(5H-imidazo[5,1-a]isoindol-5-yl)ethanol (NLG919), GDC-0919, F001287, PF-06840003, or a pharmaceutically acceptable salt or prodrug thereof, or a combination thereof.

[0164] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is 1-methyl-D-tryptophan (1-MT) (indoximod). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is tryptophan.

[0165] In some embodiments, the combinations provided herein are packaged as articles of manufacture, such as kits. In some embodiments, the articles of manufacture and / or kits further comprise instructions or literature providing for administration of the engineered cells and / or tryptophan metabolism and / or kynurenine pathway modulators.

[0166] Also provided is a method of engineering immune cells to express a recombinant receptor, comprising contacting a population of cells comprising immune cells with a tryptophan metabolism and / or kynurenine pathway modulator; and introducing a nucleic acid encoding the recombinant receptor into the population of cells under conditions such that the recombinant receptor is expressed.

[0167] In some embodiments, the recombinant receptor binds to a ligand, optionally an antigen. In some embodiments, the recombinant receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR). In some embodiments, the population of cells is or comprises peripheral blood mononuclear cells. In some embodiments, the population of cells is or comprises T cells. In some embodiments, the T cells are CD4+ and / or CD8+. In some embodiments, the population of cells is isolated from a subject, optionally a human subject.

[0168] In some embodiments, the contacting occurs before and / or during the introducing.

[0169] In some embodiments, tryptophan metabolism and / or kynurenine pathway modulators prevent or reduce the formation, concentration, availability, metabolism, and / or action of kynurenine pathway enzymes and / or metabolites. In some embodiments, tryptophan metabolism and / or kynurenine pathway modulators prevent or reduce the catabolism of tryptophan or prevent or reduce the synthesis or accumulation of tryptophan metabolites. In some embodiments, the tryptophan metabolite is selected from kynurenine, 3-hydroxykynurenine, and 3-hydroxyanthranilic acid (3-HAA).

[0170] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a small molecule, peptide, protein, antibody or antigen-binding fragment thereof, antibody mimetic, aptamer, or nucleic acid molecule.

[0171] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an antagonist of the enzyme kynureninase.

[0172] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of one or more enzymes selected from the group consisting of inhibitors of indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1), IDO2, and inhibitors of tryptophan 2,3-dioxygenase (TDO). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a dual-acting IDO / TDO inhibitor. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1).

[0173] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is 1-methyl-D-tryptophan (1-MT) (indoximod), 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N′-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat), 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-( 3-chloro-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[4-fluoro-3-(trifluoromethyl)phenyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N'-hydroxy-N-[3-(trifluoromethyl)phenyl] -1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-cyano-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[(4-bromo-2-furyl)methyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 4-({2 -[(aminosulfonyl)amino]ethyl}amino)-N-[(4-chloro-2-furyl)methyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide, 1-cyclohexyl-2-(5H-imidazo[5,1-a]isoindol-5-yl)ethanol (NLG919), GDC-0919, F001287, PF-06840003, or a pharmaceutically acceptable salt or prodrug thereof, or a combination thereof.

[0174] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is 1-methyl-D-tryptophan (1-MT) (indoximod). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is tryptophan.

[0175] Also provided are methods of engineering immune cells that express a recombinant receptor, comprising: introducing a first nucleic acid sequence encoding the recombinant receptor into a population of cells under conditions such that the recombinant receptor is expressed; and introducing one or more second nucleic acid sequences encoding agents capable of or involved in modifying expression of molecules associated with IDO-mediated immunosuppressive signaling in the cells, associated with sensing or responding to tryptophan starvation or deficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression into the population of cells under conditions such that the agents are expressed.

[0176] Also provided is a method of engineering immune cells to express a recombinant receptor, comprising introducing a first nucleic acid sequence encoding the recombinant receptor into a population of cells under conditions such that the recombinant receptor is expressed; and introducing one or more second nucleic acid sequences encoding a recombinant, engineered, and / or ectopically expressed amino acid transporter or chain thereof, or a functional and / or catalytically active portion or variant thereof.

[0177] In some embodiments, the modification comprises recombinant, engineered, and / or ectopic expression of the molecule or a functionally and / or catalytically active strand, portion, or variant thereof. In some embodiments, the modification comprises reduced expression of the molecule. In some embodiments, the method comprises an inhibitory nucleic acid or gene disruption that reduces expression of the molecule.

[0178] In some embodiments, the first nucleic acid sequence and the second nucleic acid sequence(s) are comprised in one or more polynucleotides. In some embodiments, the first nucleic acid sequence and the second nucleic acid sequence(s) are comprised in two polynucleotides. In some embodiments, the first nucleic acid sequence and the second nucleic acid sequence(s) are comprised in one or more vectors, which are optionally viral vector(s).

[0179] In some embodiments, the combinations provided herein are packaged as kits. In some embodiments, the kits also include instructions for administering the engineered cells and the tryptophan metabolism and / or kynurenine pathway modulators. In some embodiments, products and / or kits are provided, such as those containing tryptophan metabolism and / or kynurenine pathway modulators, e.g., in therapeutically effective doses. In some aspects, the products further include instructions and / or literature detailing the administration of the modulator(s) to a subject who has or will be receiving a therapy, such as a cell therapy, e.g., engineered T cell therapy. In some aspects, the instructions or literature further detail administration to a subject with a disease or condition, wherein the disease or condition or associated cells or tissues are or have been confirmed to express one or more amino acid transporters, such as IDO and / or LAT-1, optionally after administration of the therapy. In some embodiments, the instructions or literature do not indicate expression of IDO and / or do not indicate expression of transporters or other markers as a prerequisite for administration of the cell therapy. [Brief explanation of the drawings]

[0180] [Figure 1A]1 shows the expression levels of indoleamine-pyrrole 2,3-dioxygenase (IDO1) detected by flow cytometry in CD19-expressing A549 adenocarcinoma cells (CD19.A549) after stimulation with interferon gamma (IFNγ) for 24 or 48 hours, and in unstimulated CD19.A549 cells, as described in Example 1. [Figure 1B] 1 shows IDO1 expression levels detected by flow cytometry in HCC1806 human breast cancer cell line, H2286 human small cell lung cancer cell line, H1975 human non-small cell lung cancer cell line, H1299 human non-small cell lung cancer lymph node metastasis cell line, BT-549 human breast cancer cell line, and CD19-expressing A549 adenocarcinoma cells (A549.CD19) after stimulation with interferon gamma (IFNγ). [Figure 1C] 1 shows IDO1 expression levels detected by flow cytometry in Daudi cells, CD19-expressing K562 cells (K562.CD19), CD19-expressing A549 cells (A549.CD19), the BT-549 human breast cancer cell line, and HS-5 human bone marrow stromal cells after stimulation with interferon gamma (IFNγ). [Figure 1D] 1 shows IDO1 expression levels detected by flow cytometry in CD19-expressing A549 adenocarcinoma cells (CD19.A549) after co-culture with anti-CD19 chimeric antigen receptor (CAR)-expressing T cells or with anti-CD19 chimeric antigen receptor (CAR)-expressing T cells together with 10 μg / mL of anti-IFNγ antibody for either 24 or 48 hours, compared to CD19.A549 cells cultured alone, as described in Example 1. [Figure 1E] 1 is a bar graph showing the mean fluorescence intensity (MFI) of IDO1 in co-cultures with anti-CD19 chimeric antigen receptor (CAR)-expressing T cells or in co-cultures with anti-CD19 chimeric antigen receptor (CAR)-expressing T cells together with 10 μg / mL of anti-IFNγ antibody compared to CD19.A549 cells cultured alone. [Figure 1F]Figure 1 shows the mean fluorescence intensity (MFI) of IDO1 assessed by flow cytometry in A549.CD19 target cells after co-culture with anti-CD19 chimeric antigen receptor (CAR)-expressing T cells, or with anti-CD19 CAR T cells in the absence (no treatment, no Tx) or presence of increasing concentrations (from 0.75 μg / mL to 2.0 μg / mL) of anti-interferon-gamma receptor (IFNγR) antibodies or isotype controls. [Figure 1G] IDO1 expression levels detected by flow cytometry in CD19-expressing A549 adenocarcinoma cells (A549.CD19) or parental A549 cells after 24 hours of coculture with anti-CD19 chimeric antigen receptor (CAR)-expressing T cells, anti-ROR1 chimeric antigen receptor (CAR)-expressing T cells, or mock-transduced cells are shown. [Figure 1H] IFNγ production in culture supernatants after co-culture of anti-CD19 CAR+ T cells or mock-transduced cells with CD19-expressing A549 adenocarcinoma cells (A549.CD19) or parental A549 cells is shown. [Figure 1I] 1 shows CD19 expression levels in A549 target cells expressing endogenous CD19 under the control of one of two different promoters. [Figure 1J] Figure 1 shows IFNγ production from co-cultures of A549 target cells expressing endogenous CD19 under the control of one of two different promoters with anti-CD19 CAR-T cells or a non-CAR-T cell control. [Figure 1K] IDO expression, determined by flow cytometry, is depicted in the co-cultures depicted in Figure 1J. [Figure 1L] 1 shows IDO1 expression in CD19.A549 cells cultured with various concentrations of recombinant human IFNγ. [Figure 2A]Figure 1 shows the tryptophan concentration (μM) measured by ELISA in the culture supernatant of CD19.A549 cells co-cultured with anti-CD19 CAR-expressing T cells at effector-target (E:T) ratios of 0.3:1, 1:1, or 3:1 in the presence of 0 nM, 0.1 nM, 10 nM, and 1000 nM of an IDO1 inhibitor (epacadostat), as described in Example 2. The cells were cultured, and the tryptophan levels were compared to those in fresh culture medium. [Figure 2B] Figure 1 shows kynurenine concentrations (μM) measured by ELISA in culture supernatants from cocultures of CD19.A549 cells with anti-CD19 CAR-expressing T cells in the presence of 0 nM, 0.1 nM, 10 nM, and 1000 nM epacadostat at an E:T ratio of 3:1, as described in Example 2. Cells were cultured, and kynurenine levels were compared to those in fresh culture medium. [Figure 2C] Shown are concentrations of tryptophan and kynurenine (μM) after 96 hours of co-culture of anti-CD19 CAR+ T cells with A549.CD19 IDO+ target cells in the presence of various epacadostat concentrations. [Figure 2D] The concentrations (μM) of tryptophan and kynurenine measured by ELISA in the culture supernatants of CD19.A549 cells (IDO1+) or CD19.Daudi cells (IDO1-) co-cultured with anti-CD19 CAR-expressing T cells at an effector:target (E:T) ratio of 1:1 are shown. [Figure 2E] Shown are the concentrations (μM) of tryptophan and kynurenine in the culture supernatant of A549.CD19 cells (IDO+ target cells) when cultured alone for 24 hours (untreated) or with anti-CD19 CAR-expressing T cells (α-CD19 CAR) for 24 hours. [Figure 3]Figure 1 shows T cell proliferation of anti-CD19 CAR-expressing CD4+ and CD8+ T cells after co-culture with CD19-expressing A549 target cells (CD19.A549) in the presence of 0 nM, 0.1 nM, 10 nM, and 1000 nM epacadostat at 0.3:1, 1:1, or 3:1 E:T, as described in Example 3 A. T cell proliferation was measured by flow cytometry using dilutions of CellTrace™ Violet. [Figure 4-1] Figures 4A and 4B show the mean number of divisions of CD4+ (Figure 4A) or CD8+ (Figure 4B) T cells expressing anti-CD19 CAR generated from three different healthy donors after coculture with CD19-expressing A549 target cells (CD19.A549) in the presence of 0 nM, 10 nM, 100 nM, 250 nM, 500 nM, and 1000 nM epacadostat, as described in Example 3A. [Figure 4-2] Figure 4C shows the number of CAR-T cells per well when anti-CD19 CAR-expressing T cells generated from one of three different healthy donors were co-cultured with CD19.A549 cells at an E:T ratio of 1:1 or anti-ROR1 CAR-expressing T cells generated from a different donor at an E:T ratio of 3:1 in the presence of 250 nM epacadostat for 96 hours. [Figure 5-1] Figure 5A shows T cell proliferation of anti-CD19 CAR-expressing CD4+ and CD8+ T cells after co-culture with IDO1+ A549 target cells (CD19.A549) or IDO1- Daudi target cells (CD19.Daudi) at 1:1 E:T in the presence of 0 nM, 10 nM, 100 nM, 250 nM, 500 nM, and 1000 nM epacadostat, as described in Example 3 A. T cell proliferation was measured by flow cytometry using dilutions of CellTrace™ Violet. [Figure 5-2]Figures 5B and 5C show the mean number of divisions of anti-CD19 CAR-expressing CD4+ (Figure 5B) or CD8+ (Figure 5C) T cells after coculture with IDO1+ A549 target cells or IDO1- Daudi target cells in the presence of 0 nM, 10 nM, 100 nM, 250 nM, 500 nM, and 1000 nM epacadostat, as described in Example 3A. [Figure 6]

[0033] Figure 1 shows the levels of tumor necrosis factor alpha (TNFα) (pg / ml) measured by a TNFα assay kit in the culture supernatants of CD19.A549 cells co-cultured with anti-CD19 CAR-expressing T cells in the presence of 0 nM, 10 nM, 100 nM, 250 nM, 500 nM, and 1000 nM epacadostat at an E:T ratio of 1:1, as described in Example 3B. [Figure 7] Figures 7A and 7B show CD25 surface expression levels, as detected by flow cytometry, of anti-CD19 CAR-expressing CD4+ (Figure 7A) or CD8+ (Figure 7B) T cells after coculture with IDO1+ A549 target cells or IDO1- Daudi target cells in the presence of 0 nM, 10 nM, 100 nM, 250 nM, 500 nM, and 1000 nM epacadostat, as described in Example 3B. [Figure 8-1] Figures 8A and 8B show CD25 surface expression levels, as detected by flow cytometry, of anti-CD19 CAR-expressing CD4+ (Figure 8A) or CD8+ (Figure 8B) T cells generated from three different healthy donors after coculture with CD19-expressing A549 target cells (CD19.A549) in the presence of 0 nM, 10 nM, 100 nM, 250 nM, 500 nM, and 1000 nM epacadostat, as described in Example 3B. [Figure 8-2]Figure 8C shows the area under the curve (AUC) obtained from a plot of NucRed-labeled CD19.A549 cells co-cultured with anti-CD19 CAR-expressing T cells incubated for approximately 340 hours at an E:T ratio of 0.25:1 in the presence of 0 nM, 3.9 nM, 15.6 nM, 62.5 nM, 250 nM, and 1000 nM epacadostat. [Figure 9A] Figure 1 shows IDO1 expression levels detected by flow cytometry in CD19-expressing A549 adenocarcinoma cells (CD19.A549) stimulated with or without interferon gamma (IFNγ) for 48 hours in the presence of increasing concentrations of a purine analog (fludarabine). [Figure 9B] Shown is the percentage of viable cells in cultures of CD19-expressing A549 adenocarcinoma cells (CD19.A549) stimulated with or without interferon gamma (IFNγ) for 48 hours in the presence of increasing concentrations of a purine analog (fludarabine). [Figure 9C] Shown are the % viable cells and Annexin V staining results over 96 hours for CD19.A549 cells cultured with or without IFNγ in the presence of 0 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, 200 μM or 1000 μM fludarabine, added at 48 hours. [Figure 9D] Shown is the mean fluorescence intensity (MFI) of IDO1 expression over 96 hours in CD19.A549 cells cultured with or without IFNγ in the presence of 0 μM, 6.25 μM or 12.5 μM fludarabine. [Figure 10-1] Figure 10A shows the total number of CD3+ cells per well in co-cultures of anti-CD19 CAR-expressing T cells incubated with CD19.A549 at a 1:1 E:T ratio for 96 hours in the presence of 0 nM, 10 nM, 100 nM, and 1000 nM epacadostat added at the beginning of the co-culture, or in the presence of 5 μg / mL supplemental tryptophan added to the cultures every 24 hours. [Figure 10-2]Figures 10B-10E show the amounts of cytokines from the co-cultures: GM-CSF (Figure 10B), IFNγ (Figure 10C), IL-2 (Figure 10D), and TNFα (Figure 10E). [Figure 10-3] Figures 10F and 10G show the mean fluorescence intensity (MFI) of CD25 expression in CD4+ T cells (Figure 10F) and CD8+ T cells (Figure 10G) in the co-cultures. [Figure 11A] FIG. 11A shows the expression levels of indoleamine-pyrrole 2,3-dioxygenase (IDO1) detected by flow cytometry in HS-5 human bone marrow stromal cells stimulated with 20 ng / mL of recombinant human IFNγ for 24 hours. [Figure 11B] FIG. 11B shows the mean fluorescence intensity (MFI) of IDO1 in dendritic cells (DCs) and macrophages (MΦ) upon stimulation with interferon-gamma (IFNγ) or lipopolysaccharide (LPS), as determined by flow cytometry. [Figure 12A] 1 shows the mean fluorescence intensity (MFI) of L-type amino acid transporter 1 (LAT1, which in humans is encoded by the SLC7A5 gene) detected by flow cytometry in Daudi, K562, HS-5 and A549 cells. [Figure 12B] 1 shows LAT1 expression levels detected by flow cytometry in HCC1806 human breast cancer cell line, H2286 human small cell lung cancer cell line, H1975 human non-small cell lung cancer cell line, H1299 human non-small cell lung cancer lymph node metastasis cell line, BT-549 human breast cancer cell line, and CD19-expressing A549 adenocarcinoma cells (A549.CD19) after stimulation with interferon gamma (IFNγ). [Figure 12C] The MFI of LAT1 in dendritic cells (DCs) and macrophages (MΦ) upon stimulation with interferon-gamma (IFNγ) or lipopolysaccharide (LPS) as determined by flow cytometry is shown. [Figure 13]Figure 1 shows T cell proliferation of CAR-expressing T cells after co-culture with HS-5 human bone marrow stromal cells or target cells at an E:T ratio of 1:1, or with HS-5 cells and target cells at a ratio of 1:1:1. T cell proliferation was measured by flow cytometry using dilutions of CellTrace™ Violet. [Figure 14A] Figure 1 shows T cell proliferation in individual wells of anti-CD19 CAR-expressing cells generated from three different donors after 96 hours of co-culture with A549.CD19, with 5 μg / mL tryptophan supplementation every 24 hours, or in the presence of 250 nM epacadostat. T cell proliferation was measured by flow cytometry using dilutions of CellTrace™ Violet. [Figure 14B] Figure 1 shows the number of CAR T cells in each well of anti-CD19 CAR-expressing cells generated from three different donors after 96 hours of co-culture with A549.CD19, with 5 μg / mL tryptophan supplementation every 24 hours, or in the presence of 250 nM epacadostat. T cell proliferation was measured by flow cytometry using dilutions of CellTrace™ Violet. [Figure 15A] Figure 1 shows the mean fluorescence intensity (MFI) of CellTrace™ Violet dye when labeled CD4+ CAR+ T cells were co-cultured with CD19-transduced K562 human myeloid leukemia target cells (CD19.K562) in tryptophan-depleted medium with or without the addition of various concentrations of supplemental tryptophan. Tryptophan-containing medium was used as a control (Trp control). [Figure 15B] Figure 1 shows the mean fluorescence intensity (MFI) of CellTrace™ Violet dye when labeled CD8+ CAR+ T cells were co-cultured with CD19-transduced K562 human myeloid leukemia target cells (CD19.K562) in tryptophan-depleted medium with or without the addition of various concentrations of supplemental tryptophan. Tryptophan-containing medium was used as a control (Trp control). [Figure 15C]The numbers of CD19.K562 target cells are shown. [Figure 15D] Shown are the numbers of CAR+ T cells from cultures of anti-CD19 CAR-expressing cells incubated with plate-bound anti-idiotypic agonist antibodies in tryptophan-depleted medium alone or in tryptophan-depleted medium supplemented with various concentrations of tryptophan. [Figure 15E] Shown are CAR+ T cell numbers (y-axis) from a similar study using anti-CD19 CAR-expressing T cells generated from three different donors. [Figure 16A] Shown are the number of CAR-T cells in each well in co-cultures of anti-CD19 CAR+ T cells and CD19-expressing A549 target cells (CD19.A549) incubated for 96 hours in the presence of 0 nM, 15.6 nM, 62.5 nM, 250 nM, and 1000 nM epacadostat. [Figure 16B] Shown are the number of CAR-T cells in each well from co-cultures without epacadostat, with or without supplemental tryptophan added every 24 hours, followed by an additional 96 hours of culture. [Figure 16C] Shown are the percentages of IFNγ+ cells (FIG. 16C) and IL-2+ cells (FIG. 16D) among CD4+ or CD8+ anti-CD19 CAR-expressing cells generated using T cells from two different donors co-cultured with CD19-expressing A549 target cells (CD19.A549) for 96 hours in cell culture medium in the absence of epacadostat (no treatment (no Tx)) or with 250 nM epacadostat (epacadostat), and then re-cultured with fresh CD19-expressing A549 target cells (CD19.A549) in tryptophan-depleted medium with or without tryptophan supplementation (medium from "no tx" cultures at 96 hours). [Figure 16D]Shown are the percentages of IFNγ+ cells (FIG. 16C) and IL-2+ cells (FIG. 16D) among CD4+ or CD8+ anti-CD19 CAR-expressing cells generated using T cells from two different donors co-cultured with CD19-expressing A549 target cells (CD19.A549) for 96 hours in cell culture medium in the absence of epacadostat (no treatment (no Tx)) or with 250 nM epacadostat (epacadostat), and then re-cultured with fresh CD19-expressing A549 target cells (CD19.A549) in tryptophan-depleted medium with or without tryptophan supplementation (medium from "no tx" cultures at 96 hours). [Figure 17A]

[0023] Figure 1 shows T cell proliferation of anti-CD19 CAR-expressing T cells after co-culture with CD19.Daudi target cells (IDO1- target cells that were found not to induce IDO1 expression) in the presence of 0 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, and 100 μM L-kynurenine, 3-hydroxyanthranilic acid (3-HAA), or 0 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, and 100 μM L-kynurenine and 3-HAA, respectively, at a 1:1 effector:target (E:T) ratio. T cell proliferation was measured by flow cytometry using dilutions of CellTrace™ Violet. [Figure 17B] Figure 1 shows the number of CAR+ T cells expressing anti-CD19 CAR after 96 hours of co-culture with IDO1- CD19.K562 cells in the presence of 6.25 μM to 100 μM L-kynurenine, 3-hydroxyanthranilic acid (3-HAA), or 6.25 μM to 100 μM L-kynurenine and 3-HAA, respectively. [Figure 18A]Figure 18 shows the CD3 cell counts of anti-CD19 CAR cells cultured for 24, 48, 72, or 96 hours in tryptophan-starved conditions (chemically defined, tryptophan-free medium) (24, 48, 72, or 96 hr Trp starvation), after which supplemental tryptophan was added and culture continued in the presence of tryptophan until cells were harvested at 24, 48, 72, or 96 hours. As a control, cells were cultured in tryptophan-sufficient medium (complete medium) for the respective time periods. Cytokine production of tryptophan-starved CD4 or CD8 cells is shown in Figure 18B or Figure 18C, respectively, for IFNγ, and in Figure 18D or Figure 18E, respectively, for TNFα. [Figure 18B] IFNγ cytokine production from tryptophan-starved CD4+ cells is shown in FIG. 18B. [Figure 18C] IFNγ cytokine production from tryptophan-starved CD8+ cells is shown in FIG. 18C. [Figure 18D] TNFα cytokine production in tryptophan-starved CD4+ cells is shown in FIG. 18D. [Figure 18E] TNFα cytokine production in tryptophan-starved CD8+ cells is shown in FIG. 18E. [Figure 19A] Volcano plot showing the statistical significance of differences in gene product expression (log10 adjusted p-values) between CAR+ T cells cultured under tryptophan-sufficient and tryptophan-starved conditions, along with the log2 fold change in expression of each gene product, including genes undergoing significant upregulation (right) or downregulation (left). [Figure 19B]RNAseq results, expressed as TPM (transcripts per kilobase million) values, in CAR+ T cells cultured under tryptophan-sufficient (Ctrl) and tryptophan-starved (Tryp) conditions are shown for two exemplary genes encoding members of the eIF2α / ATF4 integrated stress response pathway: ATF4 and DDIT3 (also known as CCAAT / enhancer-binding protein homologous protein (CHOP)). [Figure 19C] FIG. 1 shows results based on gene ontology analysis evaluating cellular signaling pathways found to be affected under tryptophan starvation conditions based on expression analysis, along with an exemplary schematic showing genes involved in the ATF4 pathway whose expression was altered. [Figure 20] 1 shows gating on live, non-mouse cells and flow cytometry plots for single cell suspensions isolated from an A549.CD19 cell (IDO+) tumor xenograft mouse model with (anti-CD19 CAR-T) or without (untreated) administration of anti-CD19 CAR-expressing T cells assessing CD3, CD8, and IDO expression. [Figure 21] Hematoxylin and eosin (H&E) staining and in situ hybridization (ISH) using a probe specific for mRNA encoding the anti-CD19 CAR, as well as immunofluorescence staining to assess protein expression of IDO1 and programmed death-ligand 1 (PD-L1), are shown in serial tumor biopsy sections from a human subject with diffuse large B-cell lymphoma (DLBCL) before and after a single infusion with autologous T cells expressing the anti-CD19 CAR. DETAILED DESCRIPTION OF THE INVENTION

[0181] Detailed Description Provided herein are methods for enhancing or modulating the proliferation and / or activity of immune cells in connection with immunotherapy or immunotherapeutic agents, e.g., compositions comprising cells for adoptive cell therapy, e.g., T cell therapy (e.g., CAR-expressing T cells), or T cell-binding therapeutics, e.g., bispecific or multispecific agents or antibodies capable of recruiting one or more T cells or other immune cells. In some embodiments, the immunotherapy or immunotherapeutic agent comprises one or more checkpoint modulators. In some embodiments, the method comprises administering a combination therapy of an immunotherapy, such as T cell therapy (e.g., CAR-expressing T cells) or T cell-binding therapy, with a tryptophan metabolic pathway modulator, such as a kynurenine pathway modulator. In some embodiments, the immunotherapy is T cell therapy, and the provided combination therapy with a tryptophan metabolic pathway modulator, such as a kynurenine pathway modulator, enhances or modulates the proliferation or activity of T cells in the T cell therapy. In some embodiments, the enhancement of proliferation or activity is to a level that repairs or restores a baseline or steady-state level of such activity, e.g., to a level or activity that would be observed in the absence of immunosuppressive effects mediated by metabolism, e.g., catabolism, of the essential amino acid tryptophan (TRP).

[0182] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of the enzyme indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1). In some embodiments, the immunotherapy, such as T cell therapy (e.g., CAR-expressing T cells) or T cell binding therapy, is a recombinant receptor-expressing T cell therapy, which is optionally a chimeric antigen receptor (CAR)-expressing cell therapy, a tumor-infiltrating lymphocyte (TIL) therapy, or a transgenic TCR therapy. In some cases, the recombinant receptor is a chimeric antigen receptor (CAR). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of the enzyme indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1), an inhibitor of IDO2, and / or an inhibitor of tryptophan 2,3-dioxygenase (TDO). In some embodiments, the modulator is an inhibitor of IDO1, which is optionally epacadostat.

[0183] All publications, including patent documents, scientific articles, and databases, referred to herein are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication were individually incorporated by reference. To the extent that a definition set forth herein conflicts or otherwise contradicts a definition set forth in a patent, application, published application, or other publication incorporated herein by reference, the definition set forth herein shall control over the definition incorporated herein by reference.

[0184] For clarity of disclosure, and not by way of limitation, the detailed description is divided into the following subsections: The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0185] I. Modulation of the Kynurenine Pathway in Therapeutics, Including Immunotherapy Provided herein is a combination therapy that includes administration of an immunotherapy, such as a cell therapy, and administration of a modulator of the tryptophan metabolic pathway, such as an alternative metabolic pathway, such as a modulator of the kynurenine pathway.

[0186] In some aspects, in the tumor microenvironment (TME) of many types of cancer, metabolism, e.g., catabolism, of the essential amino acid tryptophan (TRP) is involved in maintaining an immunosuppressive environment. In some aspects, in the TME of many types of cancer, metabolism, e.g., catabolism, of the essential amino acid tryptophan (TRP) is involved in maintaining an immunosuppressive environment. For example, in some aspects, one or more alternative tryptophan metabolic pathways are involved. Among the pathways involved in TRP catabolism is the kynurenine pathway, in which indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1) is involved in TRP degradation by converting L-tryptophan to N-formyl-L-kynurenine, which is then metabolized through a series of steps to form nicotinamide adenine dinucleotide (NAD). Several cell types in the TME, including tumor cells and certain subsets of dendritic cells (DCs), macrophages, and immature monocytes, express increased levels of IDO1 in response to inflammatory cues such as interferon-γ (IFNγ), tumor necrosis factor-α (TNFα), or activation stimuli of signal transduction and activator of transcription 3 (STAT3). In some aspects, "bystander" cells in the tumor microenvironment, e.g., stromal cells such as bone marrow stromal cells, can also express increased levels of IDO1 in response to inflammatory cues such as IFNγ or TNFα.

[0187] In some aspects, cancer or tumor cells in the TME may promote adaptive immune tolerance, a process in which cancer or tumor cells change their phenotype in response to cytotoxic or proinflammatory immune responses to limit and evade anti-tumor immune responses. In some aspects, specific recognition of cancer cells by immune cells, such as T cells or adoptively transferred T cells, can result in the production of immune-activating cytokines, such as IFNγ or TNFα. Upon cytokine production, cancer or tumor cells then respond by limiting or evading the immune response, for example, by expressing IDO1, depleting tryptophan in the microenvironment, or expressing programmed death-ligand 1 (PD-L1) by cancer cells (see, e.g., Ribas, A. (2015) Cancer Discov. 5(9):915-919). In some cases, the adaptive immune tolerance response can limit the activity of T cells in the TME through tryptophan catabolism, resulting in local amino acid depletion and the accumulation of tryptophan catabolites.

[0188] In some aspects, certain tumor or cancer cells, or other cells in the TME, may exhibit high levels of expression or upregulate the expression of an amino acid transporter, e.g., a tryptophan transporter such as LAT1 / CD98hc or LAT2 / CD98hc, e.g., in response to tryptophan starvation or exposure to a tryptophan (or other amino acid)-starved environment. In some embodiments, such upregulation and / or expression allows for the uptake or increased uptake of an amino acid, e.g., tryptophan, by the tumor cells from the environment. In some aspects, such uptake or increased uptake by tumor cells can promote or enhance a tryptophan-depleted or tryptophan-starved environment or condition in the TME (see, e.g., Broer et al. (2011) Biochem. J. 436, 193-211; Wang et al. (2015) Am J Cancer Res 5(4):1281-1294; Ribas, A. (2015) Cancer Discov. 5(9):915-919). In some aspects, such increased expression and uptake or uptake can exacerbate the effects of an already-starved environment, e.g., tumor or tumor-associated cells that have upregulated or expressed the transporter take up most or all of the available tryptophan, and T cells and / or other immune cells, e.g., therapeutically engineered T cells, are unable to compete for tryptophan and / or experience the effects of tryptophan starvation.

[0189] In some aspects, as described herein, the occurrence or aggravation of tryptophan depletion or tryptophan starvation by diseased cells and / or associated cells or tissues, for example, by increased expression and / or activity of IDO1 or by upregulation of the expression of tryptophan transporters, such as LAT1 / CD98hc or LAT2 / CD98hc, can inhibit or prevent one or more actions of cell therapy and / or immune cells, such as the activity, proliferation, and / or expansion of anti-tumor immune cells, for example, T cells. In some aspects, the activity of the kynurenine pathway and / or tryptophan metabolism in the tumor microenvironment (TME) can result in one or more immunosuppressive activities. In some aspects, IDO1 expression by tumor cells, bone marrow cells, and / or stromal cells in the tumor microenvironment (TME) contributes to an immunosuppressive state in the TME. High levels of IDO1 expression in the TME can result in a decrease or depletion of TRP and an increase in kynurenine (KYN), and inhibit anti-tumor immune responses. In some aspects, the expression of IDO1 in TME can result in the depletion of factors or metabolites required for the activity, function, proliferation, survival and / or persistence of immune cells, such as T cells. In some aspects, IDO1-expressing cells can exert a wide range of robust immunosuppressive effects, for example, in TME, by (1) directing the suppression of the effector proliferation and function of cytotoxic T lymphocytes, NK cells and plasma cells; (2) promoting the conversion of naive CD4+ T cells into CD4+CD25+FOXP3+ Tregs and activating them; and / or (3) inducing immunosuppressive activity in neighboring IDO1-expressing DCs, for example, through a process known as bystander suppression (Vacchelli et al., (2014) OncoImmunology 3:10, e957994). In some aspects, kynurenine (KYN) and its derivatives can bind to aryl hydrocarbon receptor (AHR). KYN binding to AHR results in reprogramming of naive CD4+ T helper (Th) cell differentiation, promoting the Treg phenotype while suppressing differentiation into interleukin-17 (IL-17)-producing Th (Th17) cells.Activation of AHR can also promote a tolerogenic phenotype in dendritic cells (DCs), suppressing CD8+ T cell responses and resulting in tolerance or anergy to tumor antigens and antigen-induced cell death. Accumulation of KYN and its derivatives can also be cytotoxic to CD8+ T cells, NK cells, and NK T cells.

[0190] In some cases, IDO1 is chronically activated in some cancer patients, and increased IDO1 expression can be an independent prognostic variable for decreased survival in some cancer patients, such as those with acute myeloid leukemia (AML), small cell lung cancer, melanoma, ovarian cancer, colorectal cancer, pancreatic cancer, and endometrial cancer (see, e.g., Moon et al., Journal for ImmunoTherapy of Cancer (2015) 3:51; Platten et al., Front Immunol. 2015 Jan 12;5:673; Weinmann, H., ChemMedChem 2016,11:450-466). Inhibition of IDO1 can, in some embodiments, reduce or reverse the immunosuppressive and / or other effects of IDO1 in the TME, such as depleting immune effector cells of TRPs and promoting the accumulation of KYN and its derivatives (e.g., 3-hydroxykynurenine, kynurenic acid, and 3-hydroxyanthranilic acid, some of which can be cytotoxic to CD8+ T cells, NK cells, and NK T cells, and can also promote the differentiation of CD4+ T cells into regulatory T cells (Tregs)).

[0191] In some aspects, T cells can be sensitive to the activity of the kynurenine pathway and / or tryptophan metabolism. In some aspects, T cells are sensitive to tryptophan depletion in the environment, for example, in the TME. In some cases, T cells can sense low tryptophan (TRP) levels via uncharged tRNA, resulting in the activation of GCN2 (general control non-derepressible 2) and the initiation of an amino acid starvation or stress response that leads to cell cycle arrest, cell death, or lack of proliferation. GCN2 activation also promotes the differentiation of regulatory T cells (Tregs), enhancing Treg activity and further immunosuppression. In some cases, the kynurenine pathway and / or tryptophan metabolism can induce the depletion, anergy, or cell death of effector T cells, suppression of antitumor immune responses or tolerance to tumor antigens, and the generation and activation of Tregs, contributing to an immunosuppressive state in the TME (see, e.g., Moon et al., Journal for ImmunoTherapy of Cancer (2015) 3:51; Platten et al., Front Immunol. 2015 Jan 12; 5:673; Weinmann, H., ChemMedChem 2016, 11:450-466). In some aspects, IDO1 expression in the TME can result in the production of factors or metabolites that contribute to depletion, hyporesponsiveness (anergy), lack of proliferation, terminal differentiation, and / or differentiation into a suppressive state. In some aspects, the TME can induce T cell depletion, which can lead to a gradual loss of T cell function and / or cell depletion.

[0192] In some aspects, cancer cells or tumor cells can increase the expression of amino acid transporter to increase the uptake of amino acids from TME.For example, in some aspects, the expression of components of L-type amino acid transporter (LAT), which can mediate the uptake and exchange of amino acids into and out of cells, can be increased in various cancer cells (and / or can be upregulated by tumors and / or cancer cells or tumor-associated cells in response to amino acid starvation or other TME conditions), thereby regulating mTORC1 signal transduction and protein synthesis.Increased expression of transporters such as LAT can cause the depletion or further depletion of amino acids, such as tryptophan, in TME, thereby creating a low amino acid environment.

[0193] In some aspects, the activity of immunotherapy, e.g., T cell therapy, can be limited by immunosuppressive activities or factors present in the local microenvironment of the disease or disorder, e.g., the TME. In some aspects, the TME contains or generates factors or conditions that can suppress the activity, function, proliferation, survival, and / or persistence of T cells administered for T cell therapy.

[0194] In some aspects, provided embodiments are based, at least in part, on the findings described herein that T cells, such as CAR T cells, are sensitive to tryptophan starvation. Upregulation of IDO1 in cells in the TME, including tumor cells, can create a tryptophan-starved environment, which in turn can inhibit CAR T cell survival and / or activity. In some aspects, the suppressive effects of tryptophan starvation can be reduced, overcome, and / or reversed by inhibiting factors associated with essential amino acid tryptophan (TRP) metabolism, e.g., catabolism, such as IDO1, replenishing tryptophan, and / or increasing tryptophan uptake by CAR T cells.

[0195] In some aspects, prolonged tryptophan starvation in CAR T cells can affect the ability of CAR T cells to recover from subsequent starvation under conditions in which tryptophan is replenished and / or conditions in which the conditions leading to tryptophan starvation are reversed or attenuated. In some cases, prolonged tryptophan starvation, such as prolonged tryptophan depletion in the TME, can result in a delay or inability to recover CAR T cells, for example, a delay or cessation of proliferation and / or functional activity. In some embodiments, provided embodiments can be used to prevent or reduce prolonged tryptophan starvation and / or to promote recovery of CAR T cells from tryptophan starvation. In some embodiments, provided embodiments can be used to prevent or reduce prolonged tryptophan starvation and / or to promote recovery of CAR T cells from tryptophan starvation. In some aspects, provided embodiments allow the administered CAR T cells to restore or restore T cell functional activity and / or proliferation to baseline or steady state levels, such as levels of functional activity and / or proliferation in normal or steady state and / or non-tryptophan depleted conditions.

[0196] T cell-based therapies, such as adoptive T cell therapy (including those involving the administration of cells expressing chimeric receptors specific for a disease or disorder of interest, such as chimeric antigen receptors (CARs) and / or other recombinant antigen receptors, as well as other adoptive immune cell and adoptive T cell therapies), can be effective in treating cancer and other diseases and disorders. In certain situations, available approaches to adoptive cell therapy may not always be entirely satisfactory. In some situations, optimal activity or outcome may depend on the ability of the administered cells to recognize and bind to a target, e.g., a target antigen, and to migrate, localize, and successfully enter the appropriate site within the subject, tumor, and its environment. In some situations, optimal activity or outcome may depend on the ability of the administered cells to activate, expand, exert various effector functions including cytotoxic killing and secretion of various factors such as cytokines, persist, including long-term, differentiate, transition to, or participate in reprogramming to specific phenotypic states (such as long-lived memory, less differentiated, and effector states), avoid or reduce immunosuppressive states in the local disease microenvironment, provide effective and robust recall responses after clearance and re-exposure to the target ligand or antigen, and avoid or reduce exhaustion, anergy, peripheral tolerance, terminal differentiation, and / or differentiation to suppressive states.

[0197] In some aspects, provided embodiments are based on the finding that, when stimulated by an antigen, CAR T cells can induce the expression of functional indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1) in cells, such as tumor cells, and that the functional outcome of this increase (e.g., by regulating the kynurenine pathway of tryptophan metabolism) can be modulated by inhibitors of the enzyme. In addition to tumor cells, IDO1 can be expressed by endothelial cells, mesenchymal stromal cells, such as bone marrow stromal cells, fibroblasts, and various bone marrow-derived antigen-presenting cells, such as dendritic cells (DCs) and macrophages, which may also be present in the tumor microenvironment. In some aspects, the induction of IDO1 expression in such cells can be direct or indirect, such as through the production of interferon-gamma (IFNγ) or tumor necrosis factor alpha (TNFα) in or from CAR-T cells after antigen stimulation.

[0198] In some aspects, the findings herein indicate that upon administration of CAR T cells in vivo, tumor cells and other cells in the TME can express increased levels of IDO1. In some aspects, increased IDO1 expression by tumor cells and other cells is in response to inflammatory cues such as interferon gamma (IFNγ). In some embodiments, CAR-expressing T cells can induce IDO1 expression in CAR target antigen-positive cells, which induction is mediated in part by IFNγ.

[0199] Accordingly, some embodiments provided herein are based on findings herein showing that, in some cases, antigen-specific stimulation of T cells, such as recombinant receptor-expressing T cells (e.g., CAR-T cells), can result in upregulated expression of IDO1 in one or more cells that may be found in or associated with a tumor or tumor microenvironment. Such immunosuppressive effects in certain circumstances can negatively regulate certain activity, persistence, proliferation, or other functional characteristics of a cell therapy, such as, for example, recombinant receptor-expressing T cells (e.g., CAR-T cells).

[0200] In some embodiments, administration of a tryptophan metabolism or kynurenine pathway modulator, such as an IDO1 inhibitor, in combination with cell therapy enhances the effector function (e.g., cytokine secretion), expansion, and / or persistence of the cells of the cell therapy. In some embodiments, the enhancement of proliferation or activity is to a level that restores or restores the baseline or steady-state level of such activity, for example, to a level or activity that would be observed in the absence of immunosuppressive effects mediated by the metabolism, e.g., catabolism, of the essential amino acid tryptophan (TRP). In some aspects, the enhancement or increase in effector function (e.g., cytokine secretion), expansion, and / or persistence of the cells of the cell therapy occurs without an increase (and / or a decrease) in one or more markers induced upon cell activation, for example, a T cell activation marker such as CD25. In some embodiments, the cells exhibit substantial expansion and / or effector function, yet exhibit a less differentiated or less activated surface phenotype.

[0201] In some embodiments, T cells are sensitive to a tryptophan-depleted or tryptophan-starved environment, for example, if T cell activity, proliferation, and / or expansion can be inhibited in a tryptophan-starved or tryptophan-deficient environment. Tumor or cancer cells or related cells, such as associated stromal or bone marrow cells, can, in some aspects, create, cause, or exacerbate such an immunosuppressive environment or condition by upregulating the activity and / or expression of IDO1 and / or increasing the activity and / or expression of a tryptophan transporter, such as LAT1 / CD98hc or LAT2 / CD98hc. In some aspects, upregulation of the transporter can be in response to a tryptophan or other amino acid-starved environment. In some aspects, it can cause tumor or other cells to increase tryptophan or other amino acid uptake in their TME, which in some situations can deplete or reduce the supply or level of tryptophan available to immune cells, such as cells in adoptive cell therapy, for example, in an already starved environment. In some cases, cells in the TME, such as bystander cells such as stromal cells, can also upregulate IDO1 activity and / or expression. In some cases, the activity, proliferation, and / or expansion of T cells is substantially affected by the presence of cells that exhibit high expression of tryptophan transporters, such as LAT1 / CD98hc.

[0202] The results described herein are consistent with the interpretation that, in embodiments, CAR-expressing T cells administered for T cell therapy may be particularly sensitive to low TRP levels and / or kynurenine accumulation, and / or may be particularly vulnerable to such effects, for example, due to their ability to promote pathways that lead to tryptophan starvation and / or IDO upregulation, for example, upon administration and activation. The immunosuppressive effects of the TME, such as those induced by the presence and / or activity of CAR T cells and / or inflammatory signals, can be overcome by administering a regulator of a tryptophan metabolic pathway, such as an alternative metabolic pathway, for example, a regulator of the kynurenine pathway, for example, an inhibitor of IDO1.

[0203] In some aspects, the immunosuppressive effects of the TME can be overcome and / or further overcome by engineering CAR T cells to compete or survive in a tryptophan-depleted environment, e.g., by overexpression of tryptophan transporters, and / or to be less sensitive to tryptophan depletion, e.g., by reducing the expression of molecules associated with sensing or responding to tryptophan starvation or deficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression, including molecules involved in IDO-mediated or IDO-induced or tryptophan starvation-induced immunosuppressive signaling, e.g., molecules positively or negatively regulated by IDO-mediated catabolism, e.g., tryptophan depletion or accumulation of kynurenine or another tryptophan metabolite. Inhibiting portions of the kynurenine pathway that contribute to the immunosuppressive state of the TME, for example, by inhibiting TRP depletion and / or the production of KYN and its derivatives, or engineering cells to be more resistant to tryptophan-starved or tryptophan-depleted immunosuppressive environments, may increase one or more post-administration parameters or outcomes following administration of immunotherapy, such as T cell therapy (e.g., CAR-expressing T cells) or T cell conjugate therapy.

[0204] Thus, combination therapy using immunotherapy, such as T cell therapy (e.g., CAR-expressing T cells) or T cell conjugate therapy, with a tryptophan metabolism and / or kynurenine pathway modulator can reduce or reverse some of the immunosuppressive effects of the TME and tryptophan-depleted states, as well as restore and / or increase the activity, outcome, response, and / or durability of the immunotherapy, e.g., cell therapy.

[0205] II. Combination Therapy Methods for combination therapy are provided, including administering to a subject (1) a regulator of an alternative tryptophan metabolic pathway, such as a kynurenine pathway regulator, e.g., a tryptophan or tryptophan metabolism regulator, such as an inhibitor of IDO1, and (2) an immunotherapy or immunotherapeutic agent, such as adoptive immune cell therapy, e.g., T cell therapy (e.g., CAR-expressing cells, e.g., T cells) or T cell binding therapy or immunomodulatory therapy, e.g., a multispecific T cell-recruiting antibody and / or a checkpoint inhibitor. Also provided are combinations and products, such as kits, containing compositions containing immunotherapeutic agents, such as cells, and compositions containing kynurenine pathway regulators, as well as the use of such compositions and combinations to treat or prevent diseases, conditions, and disorders, including cancer. In some embodiments, the immunotherapy is cell therapy, e.g., immune cell therapy, e.g., cell therapy containing cells containing one or more recombinant immune receptors, e.g., T cell therapy (e.g., administration of CAR-expressing T cells). Such methods may include administering a modulator, e.g., a kynurenine pathway modulator, prior to, concurrently with, during, during the course of (including once during and / or periodically during) the administration, and / or after administration (e.g., initiation of administration) of an immunotherapeutic agent or immunotherapy, such as T cell therapy (e.g., CAR-expressing T cells) or other therapy, such as T cell combination therapy. In some embodiments, administration may include continuous or intermittent administration of a modulator, e.g., a kynurenine pathway modulator, and / or an immunotherapy or immunotherapeutic agent, e.g., T cell therapy. In some embodiments, the kynurenine pathway modulator is administered prior to, concurrently with, and / or after initiation of administration of T cell therapy.

[0206] In some embodiments, immunotherapy, such as T cell therapy (for example, CAR-expressing T cells) or T cell combination therapy, and tryptophan metabolism and / or kynurenine pathway regulators are provided as pharmaceutical compositions for administering to subjects.In some embodiments, the pharmaceutical composition contains one or both of the therapeutically effective amounts of agents for combination therapy, such as T cells and IDO1 inhibitors for adoptive cell therapy.In some embodiments, the agents are formulated for administration in separate pharmaceutical compositions.In some embodiments, any pharmaceutical composition provided herein can be formulated into a dosage form suitable for each administration route.

[0207] In some embodiments, a combination therapy comprising administering an immunotherapy (e.g., a T cell therapy comprising engineered cells, such as CAR-T cell therapy) and a tryptophan metabolism and / or kynurenine pathway modulator, or a composition thereof, is administered to a subject or patient having or at risk of having a disease or condition (e.g., cancer) to be treated. In some aspects, the method treats, e.g., ameliorates, one or more symptoms of the disease or condition, such as by reducing tumor burden in cancers that express an antigen recognized by the engineered T cells.

[0208] In some embodiments, immunotherapy for use in connection with the provided methods includes T cell therapy using engineered T cells, including cells engineered with recombinant receptors (e.g., CAR-T cells). In some cases, T cells can be sensitive to the activity of the kynurenine pathway and / or tryptophan metabolism. In some aspects, T cells can be sensitive to tryptophan starvation depletion in the environment, for example, in the TME. In some cases, tumor cells and / or other cells in the TME can increase the expression of amino acid transporters such as IDO1 and / or tryptophan transporters, for example, LAT1 / CD98hc or LAT2 / CD98hc, which can reduce the amount of tryptophan available to CAR T cells in the TME by metabolizing tryptophan and / or transporting extracellular tryptophan to tumor cells and / or other cells. In some cases, tryptophan starvation or depletion can result in the depletion of effector T cells, anergy or cell death, suppression of anti-tumor immune responses or tolerance to tumor antigens, generation and activation of Tregs, and can contribute to an immunosuppressive state in the TME (see, e.g., Moon et al., Journal for ImmunoTherapy of Cancer (2015) 3:51; Platten et al., Front Immunol. 2015 Jan 12; 5:673; Weinmann, H., ChemMedChem 2016, 11:450-466). In some aspects, tryptophan starvation or depletion can reduce or inhibit the functional activity and / or survival of CAR T cells.

[0209] In some aspects, prolonged tryptophan starvation or depletion in CAR T cells can affect their ability to recover from starvation or depletion and / or reverse the effects of starvation or depletion.In some cases, prolonged tryptophan starvation or depletion, for example, prolonged exposure to tryptophan depletion in the TME, can result in the delayed or inability of CAR T cell recovery, for example, delayed or stopped proliferation and / or functional activity.In some cases, prolonged tryptophan starvation or depletion, for example, for about 48 hours, 72 hours, 96 hours, 5 days, 6 days, 7 days, 14 days, 21 days, 28 days or more, can reduce or abolish the ability of T cells to recover from or reverse the effects of starvation or depletion.

[0210] In some embodiments, provided embodiments can be used to prevent or reduce long-term tryptophan starvation or depletion, and / or to promote recovery of CAR T cells from tryptophan starvation or depletion, and / or to prevent, reduce, or reverse the immunosuppressive and / or other effects of IDO1 upregulation in the TME. In some aspects, provided embodiments can be used to reduce or prevent tryptophan starvation or depletion, and / or to restore the functional activity and / or proliferation of T cells, e.g., engineered T cells for cell therapy, to baseline or steady-state levels, e.g., functional activity and / or proliferation levels in a normal or steady-state, e.g., tryptophan-sufficient, condition. In some embodiments, prevention and / or restoration is achieved by inhibiting the activity of IDO1, preventing tryptophan depletion in the TME, and / or enhancing the ability of T cells to take up available tryptophan from the environment. In some embodiments, combination therapy as described herein can be used to prevent or reduce tryptophan starvation or depletion or promote recovery of CAR-T cells from tryptophan starvation or depletion. In some cases, provided embodiments, i.e., preventing tryptophan starvation or depletion and / or promoting recovery from tryptophan starvation or depletion, e.g., to restore or maintain baseline or steady-state T cell activity, occur before the establishment of prolonged tryptophan starvation or depletion, e.g., before T cells are unable to reverse or recover from starvation or depletion.

[0211] In some aspects, provided embodiments, e.g., combination therapy of a T cell therapy such as CAR-T and / or a tryptophan metabolism and / or kynurenine pathway modulator, can be administered to a subject who has failed, no longer responds to, and / or relapsed after other therapies. In some embodiments, provided embodiments can be used to treat a subject who has failed, become resistant to, and / or relapsed after other therapies, such as prior treatment with a tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor. In some aspects, provided embodiments, e.g., combination therapy of a T cell therapy such as CAR-T and / or a tryptophan metabolism and / or kynurenine pathway modulator, can be administered to a subject who has failed, no longer responds to, and / or relapsed after administration of an IDO inhibitor, such as epacadostat.

[0212] Provided embodiments administering a tryptophan metabolism and / or kynurenine pathway modulator can improve response to newly administered cell therapy. Because a fresh source of T cells (e.g., CAR-T cells generated by obtaining T cells from a subject, engineering them to express a recombinant receptor, activating and / or expanding them) is introduced into the subject, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, can exert its effect on the administered T cells even if unengineered T cells from the subject have failed or no longer respond to prior treatment with a tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor such as epacadostat. In some embodiments, the fresh source of cells, e.g., engineered T cells, may not be subjected to tryptophan starvation. Thus, even if a subject has failed or no longer responds to prior treatment with a tryptophan metabolism and / or kynurenine pathway modulator, the administered tryptophan metabolism and / or kynurenine pathway modulator can facilitate prevention of tryptophan starvation or depletion and / or promotion of recovery from tryptophan starvation or depletion, e.g., to restore or maintain baseline or steady-state T cell activity of a fresh source of cells, e.g., engineered T cells.

[0213] In some embodiments, the combination therapy can be administered to a subject who has failed, become resistant to, and / or relapsed after other previous therapies, such as prior administration of a combination therapy of a tryptophan metabolism and / or kynurenine pathway modulator therapy with an immunomodulatory therapy, e.g., an immune checkpoint inhibitor therapy. In some embodiments, the previous immunomodulatory therapy is not a previous T cell therapy. In some embodiments, the subject has not previously been administered T cell therapy.

[0214] In some embodiments, the subject has been administered a combination of a tryptophan pathway modulator and a checkpoint inhibitor or other immunotherapy prior to administration of the CAR-T cells. In some aspects, the subject did not show benefit from the tryptophan pathway modulator compared to administration of the immunotherapy or checkpoint inhibitor alone.

[0215] In some embodiments, a subject is identified or selected for administration of one or more of the steps of the embodiments provided herein if the subject has failed, become refractory to, and / or relapsed after a previous treatment, such as prior administration of a tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor such as epacadostat, where optionally such prior administration was in combination with an immunomodulatory therapy that is not a T cell therapy, e.g., an immune checkpoint inhibitor therapy. In some embodiments, the combination therapy can be administered to a subject who has failed, become refractory to, and / or relapsed after another previous treatment, such as prior administration of a tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor such as epacadostat, in combination with an immunomodulator, e.g., an immune checkpoint inhibitor. In some embodiments, the previous immunomodulatory therapy is not a previous T cell therapy. In some embodiments, the subject has not previously been administered T cell therapy.

[0216] In some embodiments, the disease or condition to be treated can be any in which expression of an antigen is associated with and / or involved in the pathogenesis of the disease state or disorder, e.g., causes, exacerbates, or otherwise contributes to such a disease, condition, or disorder. Exemplary diseases and conditions can include malignant tumors or cellular transformations (e.g., cancer), autoimmune or inflammatory diseases, or infectious diseases caused, for example, by bacteria, viruses, or other pathogens. Exemplary antigens, including antigens associated with various diseases and conditions that can be treated, include any of the antigens described herein. In certain embodiments, recombinant receptors expressed on the engineered cells of the combination therapy, including chimeric antigen receptors or transgenic TCRs, specifically bind to antigens associated with the disease or condition.

[0217] In some embodiments, the disease or condition is a tumor, such as a solid tumor, lymphoma, leukemia, hematologic tumor, metastatic tumor, or other cancer or tumor type.

[0218] In some embodiments, the disease or condition is an infectious disease or condition, such as, but not limited to, viral, retroviral, bacterial, and protozoal infections, immunodeficiency, cytomegalovirus (CMV), Epstein-Barr virus (EBV), adenovirus, BK polyomavirus, etc. In some embodiments, the disease or condition is an autoimmune or inflammatory disease or condition, such as arthritis, e.g., rheumatoid arthritis (RA), type I diabetes, systemic lupus erythematosus (SLE), inflammatory bowel disease, psoriasis, scleroderma, autoimmune thyroid disease, Graves' disease, Crohn's disease, multiple sclerosis, asthma, and / or a transplant-related disease or condition.

[0219] In some embodiments, the antigen associated with the disease or disorder is orphan tyrosine kinase receptor ROR1, tEGFR, Her2, L1-CAM, CD19, CD20, CD22, mesothelin, CEA, hepatitis B surface antigen, antifolate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, EGP-2, EGP-4, EPHa2, ErbB2, 3 or 4, FBP, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-1, 22R-α, IL-13R-α2, kdr, kappa light chain, Lewis Y, L1 cell adhesion molecule, MAGE-A1, mesothelin, MUC1, MUC16, PSCA, NKG2D ligand, NY-ESO-1, MART-1, gp100, oncofetal antigen, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), prostate-specific antigen, PSMA, Her2 / neu, estrogen receptor, progesterone receptor, ephrin B2, CD123, CS-1, c-Met, GD-2, MAGE A3, CE7, Wilms' tumor 1 (WT-1), cyclin such as cyclin A1 (CCNA1), and / or biotinylated molecules, and / or molecules expressed by HIV, HCV, HBV or other pathogens.In some embodiments, the antigen associated with the disease or disorder is orphan tyrosine kinase receptor ROR1, B-cell maturation antigen (BCMA), carbonic anhydrase 9 (CAIX), tEGFR, Her2 / neu (receptor tyrosine kinase erbB2), L1-CAM, CD19, CD20, CD22, mesothelin, CEA, hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimer, EGFR vIII, folate-binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-α, IL-13R-α2, kinase insert domain receptor (kdr), kappa light chain, Lewis Y, L1 cell adhesion molecule (L1-CAM), melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, melanoma preferentially expressed antigen (PRAME), survivin, TAG72, B7-H6, IL-13 receptor α2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250 / CAIX, HLA-AI MAGE A1, HLA-A2 NY-ESO-1, PSCA, folate receptor a, CD44v6, CD44v7 / 8, avb6 integrin, 8H9, NCAM, VEGF receptor, 5T4, fetal AchR, NKG2D ligand, CD44v6, dual antigen, cancer-testis antigen, mesothelin, mouse CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, fetal tumor antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), H The antigen is selected from the group consisting of er2 / neu, estrogen receptor, progesterone receptor, ephrin B2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms' tumor 1 (WT-1), cyclin, cyclin A2, CCL-1, CD138, pathogen-specific antigens and antigens associated with universal tags, and / or biotinylated molecules, and / or molecules expressed by HIV, HCV, HBV or other pathogens.In some embodiments, the antigen targeted by the receptor comprises an antigen associated with a B cell malignancy, such as any of several known B cell markers, hi some embodiments, the antigen targeted by the receptor is CD20, CD19, CD22, ROR1, CD45, CD21, CD5, CD33, Igκ, Igλ, CD79a, CD79b, or CD30.

[0220] In some embodiments, the methods of the present invention can be used to treat myeloma, lymphoma, or leukemia. In some embodiments, the methods can be used to treat non-Hodgkin's lymphoma (NHL), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), acute myeloid leukemia (AML), or myeloma, such as multiple myeloma (MM). In some embodiments, the methods can be used to treat MM or DBCBL.

[0221] In some embodiments, the methods can be used to treat cancers that are not B-cell malignancies and / or that do not express B-cell antigens. In some embodiments, the methods can be used to treat cancers that do not express CD19. In some embodiments, the provided methods use recombinant receptor-expressing T cells (e.g., CAR-T cells) that do not target or specifically bind to CD19.

[0222] In some embodiments, the methods of the present invention can be used to treat non-hematological cancers, such as solid tumors. In some embodiments, the methods can be used to treat bladder cancer, lung cancer, brain cancer, melanoma (e.g., small cell lung cancer, melanoma), breast cancer, cervical cancer, ovarian cancer, colorectal cancer, pancreatic cancer, endometrial cancer, esophageal cancer, kidney cancer, liver cancer, prostate cancer, skin cancer, thyroid cancer, or uterine cancer.

[0223] In some embodiments, the disease or disorder is associated with an immunosuppressive local environment. In some embodiments, the disease or disorder is associated with tryptophan metabolism and / or kynurenine pathway activation in the local environment of the disease. For example, in some embodiments, the disease or disorder is associated with a decrease in tryptophan levels and / or an increase in the level of kynurenine (KYN) or its derivatives in the local microenvironment of the disease or disorder, such as the tumor microenvironment (TME). In some embodiments, the disease or disorder is associated with an increase in the expression of enzymes or factors involved in the kynurenine pathway, such as indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1), in cells within the TME, or cells within the TME are induced to express enzymes or factors involved in the kynurenine pathway, such as indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1).

[0224] In some embodiments, the disease or disorder is not associated with increased expression of an enzyme or factor involved in the kynurenine pathway, such as indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1). In some embodiments, the disease or disorder, e.g., cancer, comprises tumors that are negative for IDO1, IDO2, or TDO, and / or the subject has not been selected for the expression of IDO1, IDO2, or TDO-positive tumors.

[0225] For the prevention or treatment of disease, the appropriate dosage of tryptophan metabolism and / or kynurenine pathway regulator (e.g., IDO1 inhibitor) and / or immunotherapy, such as T cell therapy (e.g., CAR-expressing T cells) or T cell-binding therapy, may depend on the type of disease being treated, the type of tryptophan metabolism and / or kynurenine pathway regulator, the cells and / or recombinant receptors expressed on the cells, the severity and course of the disease, the route of administration, whether the tryptophan metabolism and / or kynurenine pathway regulator, e.g., IDO1 inhibitor, and / or immunotherapy, e.g., T cell therapy, is administered for prophylactic or therapeutic purposes, previous therapy, frequency of administration, the subject's medical history and response to the cells, and the discretion of the attending physician. In some embodiments, the composition and cells are suitably administered to the subject once or over a series of treatments. Exemplary administration regimens and schedules for the provided combination therapies are described.

[0226] In some embodiments, immunotherapy, such as T cell therapy, and tryptophan metabolism and / or kynurenine pathway modulators are administered as part of a further combined therapy, which can be administered simultaneously with another therapeutic intervention or sequentially in any order.In some situations, the cells are co-administered with another therapy close enough in time so that a group of cells enhances the effect of one or more additional therapeutic agents, or vice versa.In some embodiments, the cells are administered before one or more additional therapeutic agents.In some embodiments, the cells are administered after one or more additional therapeutic agents.In some embodiments, the method further comprises lymphodepleting therapy, such as the administration of a chemotherapeutic agent.In some embodiments, the method does not comprise lymphodepleting therapy.

[0227] Before, during, or after administration of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy) and / or tryptophan metabolism and / or kynurenine pathway modulators, the biological activity of the immunotherapy, e.g., the biological activity of an engineered cell population, is measured in some embodiments, e.g., by any of several known methods. Parameters evaluated include the ability of the engineered cells to destroy target cells, measured using any suitable method known in the art, such as the assays further described in Section III below. In some embodiments, the biological activity of cells, e.g., T cells administered for T cell-based therapy, is measured by assaying the expression and / or secretion of one or more cytokines. In some aspects, biological activity is measured by assessing disease burden and / or clinical outcomes, such as reduction in tumor burden or burden. In some embodiments, the levels and / or changes or alterations in levels of factors or effectors, e.g., kynurenine pathway enzymes and / or metabolites, are measured. In some embodiments, the administration of one or both agents of the combination therapy and / or any repeat administration of the therapy can be determined based on the results of assays conducted before, during, over the course of, or after administration of one or both agents of the combination therapy. In some embodiments, the results of such assays can be used to identify, select, screen, and / or exclude subjects before and / or after administration of one or more steps of the combination therapy, e.g., immunotherapy (e.g., T-cell therapy such as CAR-T cell therapy) and / or tryptophan metabolism and / or kynurenine pathway modulator. In some embodiments, the results of the assays are used to identify or select a particular subject for treatment with immunotherapy (e.g., T-cell therapy such as CAR-T cell therapy) and / or tryptophan metabolism and / or kynurenine pathway modulator.

[0228] A. Administration of Immunotherapy (e.g., T Cell Therapy or T Cell Conjugate Therapy) In some embodiments of the methods, compositions, combinations, kits, and uses provided herein, the combination therapy includes administering to the subject an immunotherapy, such as a T cell therapy (e.g., CAR-expressing T cells) or a T cell-binding therapy. Such therapy can be administered before, after, or simultaneously with the administration of one or more tryptophan metabolism and / or kynurenine pathway modulators (e.g., IDO1 inhibitors), as described.

[0229] In some embodiments, immunotherapy is a cell therapy that is or includes the administration of immune cells, such as T cells or NK cells, that target molecules expressed on the surface of a lesion, such as a tumor or cancer. In some embodiments, the immune cells express a T cell receptor (TCR) or other antigen-binding receptor. In some embodiments, the immune cells express a recombinant receptor, such as a transgenic TCR or chimeric antigen receptor (CAR). In some embodiments, the cells are autologous to the subject. In some embodiments, the cells are allogeneic to the subject. Examples of such cell therapies, e.g., T cell therapies, for use in the provided methods are described below.

[0230] In some embodiments, the immunotherapy is or comprises a T cell binding therapy that is or comprises a binding molecule capable of binding to a surface molecule expressed on a T cell. In some embodiments, the surface molecule is an activation component of a T cell, such as a component of the T cell receptor complex. In some embodiments, the surface molecule is CD3 or CD2. In some embodiments, the T cell binding therapy is or comprises an antibody or antigen-binding fragment. In some embodiments, the T cell binding therapy is a bispecific antibody that comprises at least one antigen-binding domain that binds to an activation component of a T cell (e.g., a T cell surface molecule, e.g., CD3 or CD2), and at least one antigen-binding domain that binds to a surface antigen on a target cell, such as a surface antigen on a tumor cell or cancer cell, e.g., any of the antigens listed herein, e.g., CD19. In some embodiments, the simultaneous or near-simultaneous binding of such an antibody to both its targets can result in a transient interaction between the target cell and the T cell, thereby resulting in T cell activation, e.g., cytotoxic activity, and subsequent lysis of the target cell. Among such exemplary bispecific antibody T cell engagers are bispecific T cell engager (BiTE) molecules, which comprise tandem scFv molecules fused by a flexible linker (see, e.g., Nagorsen and Bauerle, Exp Cell Res 317, 1255-1260 (2011)); tandem scFv molecules further comprising an Fc domain consisting of a first and a second subunit capable of stably associating, fused to each other, e.g., via a flexible linker (WO 2013026837); diabodies and derivatives thereof, including tandem diabodies (Holliger et al., Prot Eng 9, 299-305 (1996); Kipriyanov et al., J Mol Biol 293, 41-66 (1999)); dual affinity retargeting (DART) molecules, which may include diabody formats with C-terminal disulfide bridges; or triomabs, which include all-hybrid mouse / rat IgG molecules (Seimetz et al., Cancer Treat Rev 36, 458-467 (2010)).In some embodiments, the T cell engaging therapy is blinatumomab (CD3xCD19 BiTE) or AMG 330. Any such T cell engager can be used in the provided methods.

[0231] 1.T cell therapy In some aspects, the T cell therapy is or includes T cell therapy involving genetically engineered cells, such as tumor-infiltrating lymphocyte (TIL) therapy, transgenic TCR therapy, or recombinant receptor-expressing cell therapy. In some embodiments, the recombinant receptor specifically binds to a ligand, such as one associated with a disease or condition, e.g., one associated with or expressed on tumor or cancer cells. In some embodiments, the T cell therapy includes administering T cells engineered to express a chimeric antigen receptor (CAR).

[0232] In some embodiments, the provided cells express and / or are engineered to express receptors, such as recombinant receptors, including those comprising a ligand-binding domain or binding fragment thereof, as well as T cell receptors (TCRs) and components thereof, and / or functional non-TCR antigen receptors, such as chimeric antigen receptors (CARs). In some embodiments, the recombinant receptor comprises an extracellular ligand-binding domain that specifically binds to an antigen. In some embodiments, the recombinant receptor is a CAR that comprises an extracellular antigen-recognition domain that specifically binds to an antigen. In some embodiments, the ligand, such as an antigen, is a protein expressed on the surface of a cell. In some embodiments, the CAR is a TCR-like CAR, and the antigen is a processed peptide antigen, such as a peptide antigen of an intracellular protein, which, like a TCR, is recognized on the cell surface in association with a major histocompatibility complex (MHC) molecule.

[0233] Among the engineered cells, including engineered cells containing recombinant receptors, are those described in Section IV below. Exemplary recombinant receptors, including CARs and recombinant TCRs, and methods for engineering and introducing receptors into cells are described, for example, in International Publication Nos. WO 200014257, WO 2013126726, WO 2012 / 129514, WO 2014031687, WO 2013 / 166321, WO 2013 / 071154, WO 2013 / 123061, U.S. Patent Application Publication Nos. 2002131960, WO 2013287748, WO 20130149337, U.S. Patent No. 6,451,995, 7,446,190, 8,252,592, 8,339,645, 8,398,282, 7,446,179, 6,410,319, 7,070,995, 7,265,209, 7,354,762, 7,446,191, 8,324,353, and 8,479,118, and European Patent Application No. 2537416, and / or Sadelain et al., Cancer Discov. 2013 April;3(4):388-398; Davila et al. (2013) PLoS ONE 8(4):e61338; Turtle et al., Curr. Opin. Immunol., 2012 October;24(5):633-39; Wu et al., Cancer, 2012 March 18(2):160-75. In some aspects, the engineered antigen receptor includes the CAR described in U.S. Pat. No. 7,446,190 and those described in WO2014055668A1.

[0234] In some embodiments, T cell therapies for use in connection with the provided methods include engineered T cells, including cells engineered with recombinant receptors (e.g., CAR-T cells) that have altered expression, such as decreased or increased expression, of one or more molecules associated with IDO-mediated immunosuppressive signaling in cells, associated with sensing or responding to tryptophan starvation, deficiency, or insufficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression (e.g., due to accumulation of tryptophan metabolites), compared to the expression of such proteins induced during the normal course of IDO-mediated signaling and / or in response to or associated with sensing tryptophan starvation or deficiency and / or in response to or associated with sensing kynurenine-mediated immunosuppression. Among such proteins are proteins whose expression, when altered in T cells in response to tryptophan metabolism, e.g., catabolism, such as occurs as a result of IDO activity, contributes to, causes, or is associated with T cell immunosuppression, which in some cases leads to immune tolerance and tumor evasion from immune surveillance. Such proteins may include those that respond to the reduction, depletion, or deficiency of specific amino acids, such as tryptophan, and / or to the increase or accumulation of specific amino acid metabolites, such as kynurenine. Examples of such proteins include, but are not limited to, GCN2 kinase, BLIMP-1 / PRDM1, aryl hydrocarbon receptor (AHR), AHR nuclear translocator (ARNT), mTOR, and protein kinase C theta (PKC-θ).Upregulated or translocated expression of GCN2, CHOP, BLIMP-1, AHR, or AHR nuclear translocator (ARNT), and / or downregulated or decreased expression of mTOR or PKC-θ, in some cases, is associated with autophagy, T cell anergy, decreased T cell activation, and / or decreased T cell proliferation (see, e.g., Metz et al. (2012) Oncoimmunology, 1:1460-1468; Moon et al. (2015) J Immunother Cancer, 3:51; Platten et al. (2015) Frontiers in Immunology, 5:1).

[0235] In some embodiments, the provided engineered cells are modified by recombinant, engineered, and / or ectopic expression of one or more proteins whose expression is decreased or reduced in T cells under normal or natural circumstances upon IDO-mediated signaling and / or in response to tryptophan shortage or accumulation of tryptophan metabolites (e.g., kynurenine). In some embodiments, the one or more proteins are mTOR or PKC-theta. In some embodiments, the provided engineered cells are modified by recombinant, engineered, and / or ectopic expression of one or more molecules that can promote the uptake of tryptophan or other amino acids, such as one or more chains of an amino acid transporter, or one or more amino acid transporters, such as CD98 heavy chain (4F2hc; SLC3A2), L-type amino acid transporter 1 (LAT1; SLC7A5), LAT2 (SLC7A8), and / or proton-assisted amino acid transporter 4 (PAT4; SLC36A4), by cells expressing such molecules in tryptophan starvation or tryptophan depletion conditions.

[0236] In some embodiments, the engineered cells provided comprise recombinant, engineered, or ectopic expression of mTOR, PKC-theta, CD98hc, LAT1, LAT2, or PAT4, or a functional portion or variant thereof, such as by introduction of a nucleic acid construct encoding mTOR, PKC-theta, CD98hc, LAT1, LAT2, or PAT4. In some embodiments, expression of mTOR, PKC-theta, CD98hc, LAT1, LAT2, or PAT4 is under the control of a heterologous promoter or enhancer, e.g., under the control of an inducible promoter. In some embodiments, following IDO-mediated signaling and / or in response to tryptophan starvation or protein expression, such engineered cells can enhance or increase expression of mTOR, PKC-theta, CD98hc, LAT1, LAT2, or PAT4 in the cell relative to expression of the protein in a similar cell that has not been so modified by recombinant, engineered, and / or ectopic expression. In some embodiments, expression of such a protein, e.g., mTOR, PKC-theta, CD98hc, LAT1, LAT2, or PAT4, is increased by at least or at least approximately 1.2-fold, 1.5-fold, 2.0-fold, 3.0-fold, 5.0-fold, 6.0-fold, 7.0-fold, 8.0-fold, 9.0-fold, 10.0-fold or more under particular conditions associated with IDO-mediated signaling, trypphan insufficiency, and / or accumulation of tryptophan metabolites, compared to expression of the protein under the same conditions in a cell that has not been so modified by recombinant, engineered, and / or ectopic expression.

[0237] In some embodiments, the engineered cells provided are modified by reduced expression of one or more proteins whose expression is normally increased and / or translocated in T cells upon IDO-mediated signaling and / or in response to tryptophan deficiency or accumulation of tryptophan metabolites (e.g., kynurenine). In some embodiments, the one or more proteins are GCN2 kinase, BLIMP-1 / PRDM1, AHR, or AHR nuclear translocator (ARNT). In some embodiments, such cells have reduced expression of GCN2, CHOP, BLIMP-1 / PRDM1, AHR, or ARNT. In some embodiments, the reduced expression is achieved through an agent such as an inhibitory nucleic acid molecule or an agent capable of mediating gene disruption of a gene encoding the protein. In some embodiments, expression of the agent is under the control of a conditional promoter or enhancer, such as an inducible promoter, thereby conditionally regulating the reduced expression of such proteins. Exemplary systems and agents for producing reduced expression and / or gene disruption are described below. In some embodiments, expression of one or more proteins (e.g., GCN2, CHOP, BLIMP-1 / PRDM1 and / or AHR) in a cell is reduced by at least 50, 60, 70, 80, 90, or 95% compared to expression in a similar cell under the same conditions in the absence of the agent or gene disruption.

[0238] In some embodiments, the engineered cells comprise recombinant, engineered, or ectopic expression of one or more of mTOR, PKC-theta, CD98 heavy chain (4F2hc; SLC3A2), L-type amino acid transporter 1 (LAT1; SLC7A5), LAT2 (SLC7A8), and / or proton-assisted amino acid transporter 4 (PAT4; SLC36A4), or functional portions or variants thereof, and comprise reduced expression of one or more of GCN2, CHOP, BLIMP-1 / PRDM1, or AHR.

[0239] In some embodiments, the provided engineered cells, in which the expression of proteins associated with IDO-mediated signaling, such as in response to tryptophan shortage or accumulation of tryptophan metabolites, has been modified, can be used in combination with a tryptophan metabolism and / or kynurenine pathway modulator (e.g., an IDO1 inhibitor), to provide certain additional advantages over methods in which the engineered cells are not so modified. In some aspects, a particular tryptophan metabolism and / or kynurenine pathway modulator may not target, e.g., inhibit, all downstream activities associated with tryptophan catabolism, in which case the provided engineered cells can provide further resistance of the engineered T cell therapy to the immunosuppressive effects of the kynurenine pathway and / or IDO1 or other pathway enzyme signaling. In some cases, combining such modified engineered cells with a combination therapy including a tryptophan metabolism and / or kynurenine pathway modulator (e.g., an IDO1 inhibitor) can reduce the dosage or frequency of administration of the tryptophan metabolism and / or kynurenine pathway modulator (e.g., an IDO1 inhibitor). In certain aspects, the protein modifications in the provided engineered cells are conditional, e.g., inducible, such that modification of protein expression (e.g., one or more of GCN2, CHOP, BLIMP-1, ARH, ARNT, mTOR, PKC-theta, CD98hc, LAT1, LAT2, or PAT4) can be controlled under certain conditions, such as when tryptophan metabolism and / or kynurenine pathway modulators may not be available or active.

[0240] In some embodiments, the induction of altered expression (e.g., of decreased or increased expression) is at a time when there is a decrease in tryptophan levels, an increase in kynurenine levels, and / or an increase in IDO1, IDO2, or TDO expression or activity in a biological sample from the subject compared to before administration of T cell therapy begins or compared to an earlier time point after administration of T cell therapy begins. In some embodiments, the induction of altered expression (e.g., decreased or increased expression) is at a time when there is evidence of increased T cell anergy or exhaustion of T cell therapy, decreased T cell proliferation, and / or decreased T cell activation. In any such embodiment, such increase or decrease can be greater than 1.5-fold, 2.0-fold, 3.0-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, or more, or greater than about 1.5-fold, about 2.0-fold, about 3.0-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, or more, compared to the level or activity before administration of T cell therapy began, or compared to an earlier time point after initiation of T cell therapy. In some embodiments, the induction of altered expression (e.g., decreased or increased expression) occurs at a time point where the number of T cell therapy cells detectable in the blood is reduced compared to the number of T cells at an earlier time point after initiation of T cell therapy (e.g., a 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold or more decrease); 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 50-fold, 100-fold or more fewer than the peak or maximum number of T cell therapy cells detectable in the subject's blood after initiation of T cell therapy; or where less than or about less than 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.2%, or 0.1% of the T cell therapy cells are detectable in the blood at a time point after the peak or maximum level of such cells was detected in the blood.

[0241] In some embodiments, the provided methods include administering an inducer to induce altered expression (e.g., decreased or increased expression) of such proteins in engineered cells. In some embodiments, the inducer is allolactose or IPTG, tetracycline or a tetracycline derivative, or doxycycline or a doxycycline derivative. The choice of inducer depends on the specific induction system, such as a specific promoter, for controlling the expression of the protein or for controlling the expression of an agent (e.g., an inhibitory nucleic acid, a nuclease, or a nuclease system or complex) that mediates the disruption or reduction of the expression of the protein. In some embodiments, the inducer is rapamycin or another inducer that can promote chemically induced dimerization of the two subunits required for activity, such as inducible split-Cas9 (Zetche et al. (2015) Nat. Biotechnol., 33:139-142). Exemplary conditions for induction systems, such as induction systems, are described below.

[0242] The administration method of engineered cells for adoptive cell therapy is known, and can be used in connection with the provided methods and compositions.For example, adoptive T cell therapy methods are described, for example, in Gruenberg et al., U.S. Patent Application Publication No. 2003 / 0170238; Rosenberg, U.S. Patent No. 4,690,915; Rosenberg (2011) Nat Rev Clin Oncol.8(10):577-85).See, for example, Themeli et al., (2013) Nat Biotechnol.31(10):928-933; Tsukahara et al., (2013) Biochem Biophys Res Commun 438(1):84-9; Davila et al., (2013) PLoS ONE 8(4):e61338.

[0243] In some embodiments, cell therapy, e.g., adoptive T cell therapy, is performed by autologous transplantation, in which cells are isolated and / or otherwise prepared from the subject to be treated or from a sample derived from such a subject. Thus, in some aspects, the cells are derived from a subject, e.g., a patient, in need of treatment, and are administered to the same subject after isolation and processing.

[0244] In some embodiments, cell therapy, such as adoptive T cell therapy, is carried out by allogeneic transplantation, in which cells are isolated and / or otherwise prepared from a subject other than the subject who will or will ultimately receive cell therapy, such as the first subject.In such embodiments, the cells are then administered to a different subject of the same species, such as the second subject.In some embodiments, the first and second subjects are genetically identical.In some embodiments, the first and second subjects are genetically similar.In some embodiments, the second subject expresses the same HLA class or supertype as the first subject.

[0245] Cells can be administered by any suitable means. Cells are administered in a dosage regimen to achieve therapeutic effects, such as reducing tumor burden. Dosage and administration can depend in part on the administration schedule of tryptophan metabolism and / or kynurenine pathway regulator, which can be administered before, after, and / or simultaneously with the administration of T cell therapy.Various administration schedules for T cell therapy include, but are not limited to, single or multiple administrations over various time periods, bolus administration, and pulse infusion.

[0246] a. Compositions and Formulations In some embodiments, the dose of cells for T cell therapy, such as T cell therapy comprising cells engineered with a recombinant antigen receptor, e.g., a CAR or TCR, is provided as a composition or formulation, such as a pharmaceutical composition or formulation. Such compositions can be used according to the methods provided, such as in the prevention or treatment of diseases, conditions, and disorders.

[0247] In some embodiments, T cell therapy, such as engineered T cells (e.g., CAR T cells), is formulated with a pharmaceutically acceptable carrier. In some aspects, the choice of carrier is determined in part by the particular cell or agent and / or the method of administration. Accordingly, various suitable formulations exist. For example, the pharmaceutical composition may contain a preservative. Suitable preservatives may include, for example, methylparaben, propylparaben, sodium benzoate, and benzalkonium chloride. In some aspects, a mixture of two or more preservatives is used. The preservative or mixture thereof is typically present in an amount of about 0.0001% to about 2% by weight of the total composition. Carriers are described, for example, in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980). Pharmaceutically acceptable carriers are generally non-toxic to recipients at the dosages and concentrations employed and include, but are not limited to, buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol); low molecular weight compounds such as PEG-1, PEG-2, PEG-3, PEG-4, PEG-5, PEG-6, PEG-7, PEG-8, PEG-9, PEG-10, PEG-11, PEG-12, PEG-13, PEG-14, PEG-15, PEG-16, PEG-17, PEG-18, PEG-19, PEG-20, PEG-21, PEG-22, PEG-23, PEG-24, PEG-25, PEG-26, PEG-27, PEG-28, PEG-29, PEG-30, PEG-31, PEG-32, PEG-33, PEG-44, PEG-45, PEG-46, PEG-47, PEG-48, PEG-49 ... low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants such as polyethylene glycol (PEG).

[0248] In some aspects, a buffering agent is included in the composition. Suitable buffering agents include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. In some aspects, a mixture of two or more buffering agents is used. The buffering agent or mixture thereof is typically present in an amount of about 0.001% to about 4% by weight of the total composition. Methods for preparing administrable pharmaceutical compositions are known. Exemplary methods are described in more detail, for example, in Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins; 21st ed. (May 1, 2005).

[0249] Formulations or compositions may also contain multiple active ingredients useful for the particular indication, disease, or condition being prevented or treated with the cells or agents, provided that their respective activities do not adversely affect each other. Such active ingredients are suitably present in combination in amounts effective for the intended purpose. Thus, in some embodiments, the pharmaceutical composition further comprises other pharmaceutically active agents or drugs, such as chemotherapeutic agents, e.g., asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, vincristine, etc. In some embodiments, the agents or cells are administered in the form of a salt, e.g., a pharmaceutically acceptable salt. Suitable pharmaceutically acceptable acid addition salts include those derived from mineral acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, metaphosphoric acid, nitric acid, and sulfuric acid, and organic acids such as tartaric acid, acetic acid, citric acid, malic acid, lactic acid, fumaric acid, benzoic acid, glycolic acid, gluconic acid, succinic acid, and arylsulfonic acids, e.g., p-toluenesulfonic acid.

[0250] The active ingredient can be encapsulated in microcapsules, colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or macroemulsions.In certain embodiments, pharmaceutical compositions are formulated as inclusion complexes, such as cyclodextrin inclusion complexes, or as liposomes.Liposomes can help target agents or host cells (for example, T cells or NK cells) to specific tissues.Many methods are available for preparing liposomes, such as those described in Szoka et al., Ann.Rev.Biophys.Bioeng.,9:467(1980), and U.S. Patent Nos. 4,235,871, 4,501,728, 4,837,028 and 5,019,369.

[0251] In some aspects, pharmaceutical compositions can use time-release, delayed release and sustained release delivery system, so that the delivery of the composition occurs before the sensitization of the treatment site and in sufficient time to cause sensitization.Many kinds of release delivery systems are available and known.Such system can avoid the repeated administration of the composition, thereby increasing the convenience for patients and doctors.

[0252] In some embodiments, the pharmaceutical composition contains an amount of the agent or cells effective to treat or prevent a disease or condition, such as a therapeutically effective amount or a prophylactically effective amount. In some embodiments, the outcome(s) and / or effectiveness of the treatment or prevention are monitored by periodic evaluation of the treated subject. For repeated administration over several days or longer, treatment is repeated until the desired suppression or amelioration or prevention of the signs or symptoms of the disease or condition occurs, or is deemed to have occurred or is likely to occur, depending on the condition. However, other administration regimens may also be useful and can be determined. The desired dosage can be delivered by a single bolus administration of the composition, by multiple bolus administrations of the composition, or by continuous infusion administration of the composition.

[0253] The agents or cells can be administered by any suitable means, such as bolus injection, by injection, for example, intravenous or subcutaneous injection, intraocular injection, periocular injection, subretinal injection, intravitreal injection, transseptal injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjunctival injection, subconjunctival injection, sub-Tenon injection, retrobulbar injection, peribulbar injection, or posterior parascleral delivery. In some embodiments, they are administered parenterally, intrapulmonaryly, and intranasally, and, if desired for localized treatment, intralesionally. Parenteral injections include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

[0254] In some cases, cell therapy is administered as a single pharmaceutical composition comprising cells.In some embodiments, a given dose is administered by a single bolus administration of cells or agent.In some embodiments, a given dose is administered by multiple bolus administrations of cells or agent over a period of, for example, 3 days or less, or by continuous infusion administration of cells or agent.

[0255] For the prevention or treatment of disease, the appropriate dosage will depend on the type of disease being treated, the type of agent(s), the type of cells or recombinant receptor, the severity and course of the disease, whether the agent or cells are being administered for prophylactic or therapeutic purposes, previous therapy, the subject's medical history and response to the agent or cells, and the discretion of the attending physician. The compositions, in some embodiments, are suitably administered to the subject at one time or over a series of treatments.

[0256] Cells or agents can be administered using standard administration techniques, formulations, and / or devices. Formulations and devices, such as syringes and vials, are provided for storing and administering the compositions. With respect to cells, administration can be autologous or xenogeneic. For example, immunoresponsive cells or precursors can be obtained from one subject and administered to the same subject or a different, compatible subject. Peripheral blood-derived immunoresponsive cells or their progeny (e.g., derived in vivo, ex vivo, or in vitro) can be administered by catheter administration, systemic injection, local injection, intravenous injection, or local injection, including parenteral administration. When administering a therapeutic composition (e.g., a pharmaceutical composition containing genetically modified immunoresponsive cells or agents that treat or ameliorate symptoms of neurotoxicity), the therapeutic composition is generally formulated into a unit-dose injectable form (solution, suspension, emulsion).

[0257] Formulations include those for oral, intravenous, intraperitoneal, subcutaneous, pulmonary, transdermal, intramuscular, intranasal, buccal, sublingual, or suppository administration. In some embodiments, the agent or cell population is administered parenterally. As used herein, the term "parenteral" includes intravenous, intramuscular, subcutaneous, rectal, vaginal, and intraperitoneal administration. In some embodiments, the agent or cell population is administered to a subject using peripheral systemic delivery via intravenous, intraperitoneal, or subcutaneous injection.

[0258] In some embodiments, the compositions are provided as sterile liquid preparations, such as isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions, which may be buffered to a selected pH in some aspects. Liquid preparations are usually easier to prepare than gels, other viscous compositions, and solid compositions. Furthermore, liquid compositions are somewhat more convenient to administer, especially by injection. On the other hand, viscous compositions can be formulated within an appropriate viscosity range that provides a longer contact period with specific tissues. Liquid or viscous compositions can contain a carrier, which can be a solvent or dispersion medium, including, for example, water, saline, phosphate-buffered saline, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol), and suitable mixtures thereof.

[0259] Sterile injectable solutions can be prepared by incorporating the agent or cells in a solvent such as a mixture with a suitable carrier, diluent, or excipient, e.g., sterile water, saline, glucose, dextrose, etc. The composition can also be lyophilized. The composition may contain auxiliary substances such as wetting agents, dispersing or emulsifying agents (e.g., methylcellulose), pH buffering agents, gelling or thickening additives, preservatives, flavoring agents, coloring agents, etc., depending on the desired route of administration and formulation. In some aspects, standard texts may be consulted to prepare appropriate formulations.

[0260] Various additives can be added to enhance the stability and sterility of the composition, including antimicrobial preservatives, antioxidants, chelating agents, and buffers. Prevention of microbial action can be ensured by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, etc. Prolonged absorption of injectable pharmaceutical forms can be achieved by the use of agents that delay absorption, such as aluminum monostearate and gelatin.

[0261] Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, eg, films, or microcapsules.

[0262] Formulations to be used for in vivo administration are generally sterile. Sterility may be readily accomplished, for example, by filtration through sterile filtration membranes.

[0263] b. Dosage schedule and administration In some embodiments, a dose of cells is administered to a subject according to the provided methods. In some embodiments, the size or timing of the dose is determined as a function of the particular disease or condition in the subject. It is within the level of ordinary skill in the art to empirically determine the size or timing of the dose for a particular disease in light of the provided instructions.

[0264] In certain embodiments, the cells, or individual populations of cell subtypes, are present in a range of about 100,000 to about 100 billion cells, and / or that amount of cells per kilogram of subject body weight, e.g., 100,000 to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or a range defined by any two of the foregoing values), 1 million to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or a range defined by any two of the foregoing values), e.g., about 10 million to about 100 billion cells (e.g., about 20 million cells, about 30 million cells, about 40 million cells, about 60 million cells, about 70 million cells, about 800 million cells, about 90 million cells, about 10 billion cells, about 25 billion cells, about 50 billion cells, about 75 billion cells, about 90 billion cells, or a range defined by any two of the foregoing values), and in some cases, about 100 million cells to about 5 0 billion cells (e.g., about 120 million cells, about 250 million cells, about 350 million cells, about 450 million cells, about 650 million cells, about 800 million cells, about 900 million cells, about 3 billion cells, about 30 billion cells, about 45 billion cells), or any value between these ranges and / or that amount of cells per kilogram of the subject's body weight. Dosages may vary depending on the disease or disorder and / or patient and / or other treatment-specific attributes. In some embodiments, such values ​​refer to the number of recombinant receptor-expressing cells, while in other embodiments, they refer to the number of T cells or PBMCs or total cells administered.

[0265] In some embodiments, for example when the subject is a human, the dose is about 1 x 10 8 Less than 1 x 10 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs), e.g., 1 x 10 6 ~1×10 8of such cells, e.g., 2 x 10 6 , 5×10 6 , 1×10 7 , 5×10 7 , or 1×10 8 or a range of such total cells, or a range between any two of the foregoing values. In some embodiments, when the subject is a human, the dose is about 1 x 10 6 ~3×10 8 of total recombinant receptor (e.g., CAR)-expressing cells, e.g., about 1×10 7 ~2×10 8 of such cells, e.g., about 1 x 10 7 , 5×10 7 , 1×10 8 Or 1.5 x 10 8 In some embodiments, the patient is administered multiple doses, each dose or total dose may be within any of the ranges set forth above. In some embodiments, the dose of cells is between 1 x 10, ... 5 ~5×10 8 or approximately 1 x 10 total recombinant receptor-expressing T cells or total T cells 5 ~1×10 8 or approximately 5 x 10 total recombinant receptor-expressing T cells or total T cells 5 ~1×10 7 or approximately the above values ​​of total recombinant receptor-expressing T cells or total T cells, or 1 x 10 6 ~1×10 7 or administration of about the above amounts of total recombinant receptor-expressing T cells or total T cells.

[0266] In some embodiments, the cell therapy is administered at a concentration of at least or at least about, or at or about, 0.1 x 10 per kg of subject body weight. 6 cells / kg, 0.2×10 6 cells / kg, 0.3×10 6 cells / kg, 0.4×10 6 cells / kg, 0.5×10 6 cells / kg, 1×106 cells / kg, 2.0×10 6 cells / kg, 3×10 6 cells / kg or 5 x 10 6 It involves administering a dose containing a cell number of cells / kg.

[0267] In some embodiments, the cell therapy is administered at a dose of 0.1 x 10 per kg of subject body weight, inclusive. 6 cells / kg~1.0×10 7 cells / kg or approximately the above value, 0.5 x 10 6 cells / kg~5×10 6 cells / kg or approximately the above value, 0.5 x 10 6 cells / kg~3×10 6 cells / kg or approximately the above value, 0.5 x 10 6 cells / kg~2×10 6 cells / kg or approximately the above value, 0.5 x 10 6 cells / kg~1×10 6 cells / kg or approximately the above value, 1.0 x 10 6 cells / kg~5×10 6 cells / kg or approximately the above value, 1.0 x 10 6 cells / kg~3×10 6 cells / kg or approximately the above value, about 1.0 x 10 6 cells / kg~2×10 6 cells / kg or approximately the above value, 2.0 x 10 6 cells / kg~5×10 6 cells / kg or approximately the above value, 2.0 x 10 6 cells / kg~3×10 6 cells / kg or approximately the above value, or 3.0 x 10 6 cells / kg~5×10 6 This includes administering a dose containing cells / kg or about the number of cells stated above.

[0268] In some embodiments, the dose of cells is 2×10 5 cells / kg~2×10 6 cells / kg or approximately the value above, e.g., 4 x 10 5 cells / kg~1×106 cells / kg or approximately the above value, or 6 x 10 5 cells / kg~8×10 5 In some embodiments, the dose of cells is at or about 2 x 10 cells / kg per kilogram of subject body weight (cells / kg). 5 3×10 or less cells (e.g., antigen-expressing cells such as CAR-expressing cells), e.g., 3×10 5 cells / kg or less, or approximately equal to or less than 4 x 10 5 cells / kg or less, or approximately the above value, 5 x 10 5 cells / kg or less, or approximately the above value, 6 x 10 5 cells / kg or less, or approximately the above value, 7 x 10 5 cells / kg or less, or approximately equal to or less than 8 x 10 5 cells / kg or less, or approximately equal to 9 x 10 5 cells / kg or less, or approximately the above value, 1 x 10 6 cells / kg or less, or approximately equal to or less than 2 x 10 6 In some embodiments, the dose of cells is 2 x 10 cells / kg or less per kilogram of subject body weight (cells / kg). 5 cells (e.g., antigen-expressing cells such as CAR-expressing cells) or approximately 2 x 10 5 cells, or at least 2 x 10 5 cells or at least approximately 2 × 10 5 cells, for example, at or about, or at least about, 3 x 10 5 cells / kg, 4×10 5 cells / kg, 5×10 5 cells / kg, 6×10 5 cells / kg, 7×10 5 cells / kg, 8×10 5 cells / kg, 9×10 5 cells / kg, 1×10 6 cells / kg, or 2 x 10 6 Contains cells / kg.

[0269] In the context of adoptive cell therapy, administration of a given "dose" of cells encompasses administration of a given amount or number of cells as a single composition and / or as a single, uninterrupted administration, such as a single injection or continuous infusion, as well as administration of a given amount or number of cells as divided doses provided in multiple individual compositions or infusions over a specified period of no more than three days. Thus, in some situations, a dose is a single or continuous administration of a specified number of cells given or initiated at a single point in time. However, in some situations, a dose is administered in multiple injections or infusions over a period of three days or less, such as once daily for three or two days, or by multiple infusions over a one-day period.

[0270] Thus, in some aspects, the dose of cells is administered in a single pharmaceutical composition, hi some embodiments, the dose of cells is administered in multiple compositions collectively comprising the dose of cells.

[0271] The term "split dose" refers to a dose that is divided so that it is administered over more than one day. This type of administration is encompassed by the methods of the present invention and is considered a single administration. In some embodiments, the split doses of cells are administered over a period of 3 days or less in multiple compositions collectively comprising the dose of cells.

[0272] Therefore, the dose of cells can be administered as a divided dose. For example, in some embodiments, the dose can be administered to the subject over two or three days. An exemplary method for divided administration includes administering 25% of the dose on day 1, and administering the remaining 75% of the dose on day 2. In other embodiments, 33% of the dose can be administered on day 1, and the remaining 67% can be administered on day 2. In some aspects, 10% of the dose can be administered on day 1, 30% of the dose can be administered on day 2, and 60% of the dose can be administered on day 3. In some embodiments, the divided dose does not exceed three days.

[0273] In some embodiments, a dose of cells can be administered by administering multiple compositions or solutions, such as a first and a second, optionally more, each containing a portion of the dose of cells. In some aspects, multiple compositions each containing different populations and / or subtypes of cells are administered separately or independently, optionally within a period of time. For example, the populations or subtypes of cells can each be CD8 + and CD4 + The composition may comprise CD8+ T cells, and / or CD4+ enriched populations, respectively, such as CD4+ and / or CD8+ T cells, each comprising cells genetically engineered to express a recombinant receptor. In some embodiments, administering the doses comprises administering a first composition containing a dose of CD8+ T cells or a dose of CD4+ T cells, and administering a second composition containing the dose of the other of the CD4+ T cells and CD8+ T cells.

[0274] In some embodiments, administering a composition or dose, e.g., administering multiple cell compositions, comprises administering the cell compositions separately. In some aspects, the separate administrations are simultaneous or sequential in any order. In some embodiments, the dose comprises a first composition and a second composition, and the first composition and the second composition are administered 0-12 hours apart, 0-6 hours apart, or 0-2 hours apart. In some embodiments, the initiation of administration of the first composition and the initiation of administration of the second composition occur within 2 hours, 1 hour, or within 30 minutes, 15 minutes, 10 minutes, or 5 minutes of each other. In some embodiments, the initiation and / or completion of administration of the first composition and the completion and / or initiation of administration of the second composition occur within 2 hours, 1 hour, or within 30 minutes, 15 minutes, 10 minutes, or 5 minutes of each other.

[0275] In some compositions, the first composition, e.g., the first composition of the dose, comprises CD4+ T cells. In some compositions, the first composition, e.g., the first composition of the dose, comprises CD8+ T cells. In some embodiments, the first composition is administered before the second composition.

[0276] In some embodiments, the cell dose or composition comprises a defined or target ratio of CD4+ cells expressing the recombinant receptor to CD8+ cells expressing the recombinant receptor, and / or CD4+ cells to CD8+ cells, optionally about 1:1, or about 1:3 to about 3:1, e.g., about 1:1. In some aspects, administering a composition or dose having a target or desired ratio of different cell populations (e.g., a CD4+:CD8+ ratio or a CAR+CD4+:CAR+CD8+ ratio, e.g., 1:1) comprises administering a cell composition comprising one population, followed by another cell composition comprising the other population, where the administrations are at or near the target or desired ratio.

[0277] In some embodiments, the dose of cells is generally sufficient to be effective in reducing disease burden.

[0278] In some embodiments, the cells are administered at a desired dosage, which in some aspects includes a desired dose or number of cells or cell type(s) and / or a desired ratio of cell types. Thus, in some embodiments, the dosage of cells is based on the total number of cells (or number per kg body weight) and the desired ratio of individual populations or subtypes, for example, the CD4+ to CD8+ ratio. In some embodiments, the dosage of cells is based on the desired total number of cells (or number per kg body weight) in each population or each cell type. In some embodiments, the dosage is based on a combination of characteristics such as the desired number of total cells in each population, the desired ratio, and the desired total number of cells.

[0279] In some embodiments, CD8 + and CD4 +A population or subtype of cells, such as T cells, is administered at or within a tolerance of a desired dose of total cells, such as a desired dose of T cells. In some embodiments, the desired dose is a desired number of cells or a desired number of cells per unit body weight of the subject to whom the cells are administered, e.g., cells / kg. In some aspects, the desired dose is above a minimum or minimum number of cells, or above a minimum or minimum number of cells per unit body weight. In some aspects, of the total cells administered at a desired dose, individual populations or subtypes are administered at or within a tolerance of a desired production ratio (CD4 + Against CD8 + ratio, etc.) or a ratio close thereto, e.g., within a certain tolerance or error in such ratio.

[0280] In some embodiments, cells are administered at or within a tolerance of a desired dose of cells of one or more individual populations or subtypes, such as a desired dose of CD4+ cells and / or a desired dose of CD8+ cells. In some aspects, the desired dose is the desired number of cells of a subtype or population, or the desired number of such cells per unit body weight of the subject to whom the cells are administered, e.g., cells / kg. In some aspects, the desired dose is above the minimum or minimum number of cells of the population or subtype, or above the minimum or minimum number of cells of the population or subtype per unit body weight.

[0281] Thus, in some embodiments, the dosage is based on a desired fixed dose and desired ratio of total cells, and / or based on a desired fixed dose of one or more, e.g., each, of individual subtypes or subpopulations. Thus, in some embodiments, the dosage is based on a desired fixed dose or minimum dose of T cells and CD4 + Against CD8 + Based on the desired ratio of cells and / or CD4 + and / or CD8 + Based on a desired fixed or minimum dose of cells.

[0282] In some embodiments, cells are administered at or within a tolerance range for a desired production ratio of multiple cell populations or subtypes, such as CD4+ and CD8+ cells or subtypes. In some aspects, the desired ratio can be a specific ratio or a range of ratios, e.g., in some embodiments, the desired ratio (e.g., CD4 + Against CD8 + The ratio of cells) is 5:1 to 5:1 or about 5:1 to about 5:1 (or greater than about 1:5 but less than about 5:1), or 1:3 to 3:1 or about 1:3 to about 3:1 (or greater than about 1:3 but less than about 3:1), for example, 2:1 to 1:5 or about 2:1 to about 1:5 (or greater than about 1:5 but less than about 2:1, for example, 5:1, 4.5:1, 4:1, 3.5:1, 3:1, 2.5:1, 2:1, 1.9:1, 1.8:1, 1.7:1, 1.6:1, 1.5:1, 1.4:1, 1.3:1, 1.5 ... 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5, or 1:5, or approximately. In some aspects, the tolerance is within about 1%, about 2%, about 3%, about 4%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50% of the desired ratio, including any value between these ranges.

[0283] In certain embodiments, the number and / or concentration of cells refers to the number of recombinant receptor (e.g., CAR)-expressing cells. In other embodiments, the number and / or concentration of cells refers to the number or concentration of total cells, T cells, or peripheral blood mononuclear cells (PBMCs) administered.

[0284] In some aspects, the size of the dose is determined based on one or more criteria, such as the subject's response to previous treatment, e.g., chemotherapy; the disease burden in the subject, e.g., tumor mass, volume, size, or extent, extent or type of metastasis, stage of disease; and / or the likelihood or incidence of the subject developing a toxic outcome, e.g., CRS, macrophage activation syndrome, tumor lysis syndrome, neurotoxicity, and / or the host's immune response to the administered cells and / or recombinant receptor.

[0285] In some cases, the provided methods allow for the administration of lower doses of such cells to achieve the same or better therapeutic efficacy or other outcome than in methods in which the cell therapy is administered without administration of a tryptophan metabolism and / or kynurenine pathway modulator (e.g., an IDO1 inhibitor), or in which the cells do not involve alteration of expression of proteins associated with IDO-mediated immunosuppressive signaling, associated with sensing or responding to tryptophan starvation or deficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression. In some embodiments, the dose of cells administered is less than that in methods in which the cell therapy is administered without administering a tryptophan metabolism and / or kynurenine pathway modulator (e.g., an IDO1 inhibitor), or in which the cells do not involve alteration of the expression of proteins associated with IDO-mediated immunosuppressive signaling that are induced or activated in response to or associated with sensing starvation or deficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression, e.g., at least 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, or 10-fold less than that in methods in which the cell therapy is administered without administering a tryptophan metabolism and / or kynurenine pathway modulator (e.g., an IDO1 inhibitor), or in which the cells do not involve alteration of the expression of proteins associated with IDO-mediated immunosuppressive signaling, associated with sensing or responding to tryptophan starvation or deficiency, and / or associated with sensing or responding to kynurenine-mediated immunosuppression. In some embodiments, for example, the lower dose is about 5×10 per kilogram of the subject's body weight. 6 Fewer than about 4 x 10 cells, recombinant receptor (e.g., CAR)-expressing cells, T cells, and / or PBMCs, e.g., per kilogram of subject body weight 6 , 3×10 6 , 2 × 10 6 , 1×10 6 , 0.5×10 6 or 0.1 × 10 6 It includes one or fewer such cells.

[0286] In some embodiments, one or more subsequent doses of cells can be administered to the subject. In some embodiments, the subsequent doses of cells are administered more than or about 7 days, 14 days, 21 days, 28 days or 35 days after the start of administration of the first dose of cells. The subsequent doses of cells can be more than, about the same as, or less than the first dose. In some embodiments, the administration of T cell therapy, such as the administration of the first and / or second doses of cells, can be repeated.

[0287] In some embodiments, the initiation of administration of the cell therapy, e.g., the first dose of a dose of cells or a sub-dose of cells, is administered prior to (before), concurrently with, or subsequent to (following) administration of the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor.

[0288] In some embodiments, the dose of cells, or subsequent doses of cells, is administered 0 to 90 days, e.g., 0 to 30 days, 0 to 15 days, 0 to 6 days, 0 to 96 hours, 0 to 24 hours, 0 to 12 hours, 0 to 6 hours, or 0 to 2 hours, 2 hours to 30 days, 2 hours to 15 days, 2 hours to 6 days, 2 hours to 96 hours, 2 hours to 24 hours, 2 hours to 12 hours, 2 hours to 6 hours, 6 hours to 90 days, 6 hours to 30 days, 6 hours to 15 days, 6 hours to 6 days, 6 hours to 6 hours, or 6 hours after administration of the tryptophan metabolism and / or kynurenine pathway modulator is initiated or commenced. The cells may be administered for 6 to 96 hours, 6 to 24 hours, 6 to 12 hours, 12 hours to 90 days, 12 hours to 30 days, 12 hours to 15 days, 12 hours to 6 days, 12 hours to 96 hours, 12 hours to 24 hours, 24 hours to 90 days, 24 hours to 30 days, 24 hours to 15 days, 24 hours to 6 days, 24 hours to 96 hours, 96 hours to 90 days, 96 hours to 30 days, 96 hours to 15 days, 96 hours to 6 days, 6 days to 90 days, 6 days to 30 days, 6 days to 15 days, 15 days to 90 days, 15 days to 30 days, or 30 days to 90 days. In some embodiments, the dose of cells, or subsequent doses of cells, are administered after administration of the tryptophan metabolism and / or kynurenine pathway modulator has started or commenced. In some embodiments, the dose of cells is administered about or at least about or at least about 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 2 days, 3 days, 6 days, 12 days, 15 days, 30 days, 60 days, or 90 days after starting or initiating administration of the tryptophan metabolism and / or kynurenine pathway modulator.

[0289] In some embodiments, the dose of cells is administered at a time when one or more effects of the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, have been achieved. In some embodiments, the method comprises evaluating a sample from the subject after administering the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, but before administering the T cell therapy, e.g., adoptive T cell therapy, for changes in levels of metabolites in the kynurenine pathway, e.g., TRP, KYN, or a derivative of KYN, and / or expression levels of factors or effectors involved in the kynurenine pathway, e.g., enzymes, e.g., IDO1, or other phenotypes or desired outcomes described herein, e.g., those described in Section III. In some embodiments, the dose of cells is administered at a time point when the tryptophan metabolism and / or kynurenine pathway modulator (e.g., an IDO1 inhibitor) has resulted in, or is likely to result in, a change in the level of a metabolite in the kynurenine pathway, e.g., an increase in the level of TRP or a decrease in the level of KYN or a KYN derivative (e.g., an increase or decrease of at least 1.2-fold, at least 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 10-fold, or more, or at least about 1.2-fold, at least about 1.5-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 10-fold, or more), compared to the level immediately prior to administration of the tryptophan metabolism and / or kynurenine pathway modulator. Such and other parameters, and examples for determining or assessing them, are described in Section III.

[0290] In some embodiments, the initiation of administration of a cell dose of T cell therapy, e.g., adoptive T cell therapy, is administered prior to administration of a tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor. In some embodiments, the cell dose is administered at least 1 hour, at least 2 hours, at least 3 hours, at least 6 hours, at least 12 hours, at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, or at least about 5 days prior to administration of a tryptophan metabolism and / or kynurenine pathway modulator (e.g., an IDO1 inhibitor). The administration may be at least 6 days or at least about 6 days in advance, at least 7 days or at least about 7 days in advance, at least 12 days or at least about 12 days in advance, at least 14 days or at least about 14 days in advance, at least 15 days or at least about 15 days in advance, at least 21 days or at least about 21 days in advance, at least 28 days or at least about 28 days in advance, at least 30 days or at least about 30 days in advance, at least 35 days or at least about 35 days in advance, at least 42 days or at least about 42 days in advance, at least 60 days or at least about 60 days in advance, or at least 90 days in advance.

[0291] In some embodiments, the method comprises evaluating a sample from the subject after administering a cell dose of T cell therapy, e.g., adoptive T cell therapy, but prior to administering a tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, for changes in levels of metabolites in the kynurenine pathway, e.g., TRP, KYN, or a derivative of KYN, and / or expression levels of factors or effectors involved in the kynurenine pathway, e.g., enzymes, e.g., IDO1, or other phenotypes or desired outcomes described herein, e.g., those described in Section III. In some embodiments, the administration of the tryptophan metabolism and / or kynurenine pathway modulator is at a time when the prior administration of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy) has directly or indirectly caused or induced, or is likely to have caused or induced, a change in the level of a metabolite in the kynurenine pathway, such as a decrease in the level of TRP or an increase in the level of KYN or a KYN derivative (e.g., at least at least 1.2-fold, at least 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 10-fold, or more, or an increase or decrease of at least about 1.2-fold, at least about 1.5-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 10-fold, or more), compared to the level at a time point immediately prior to or after the initiation of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy). Various parameters for determining or evaluating combination therapy regimens are described in Section III.

[0292] B. Administration of Tryptophan Metabolism and / or Kynurenine Pathway Modulators The provided methods, compositions, combinations, kits, and uses involve the administration of a modulator of the kynurenine pathway, e.g., an inhibitor of the enzyme indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered prior to, after, during, simultaneously or nearly simultaneously with, continuously, and / or intermittently, administration of immunotherapy, e.g., T cell therapy, e.g., administration of T cells expressing a chimeric antigen receptor (CAR).

[0293] 1. Tryptophan metabolism and / or kynurenine pathway regulators In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor and / or antagonist that inhibits the activity of factors and / or enzymes in the kynurenine pathway. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator affects the levels of metabolites in the kynurenine pathway, e.g., causes an increase or decrease in a particular metabolite in the kynurenine pathway.

[0294] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator modulates, e.g., regulates, inhibits, induces, or activates, one or more factors or effectors, e.g., enzymes, involved in the pathway. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator modulates, e.g., regulates, inhibits, or activates, indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1), IDO2, tryptophan 2,3-dioxygenase (TDO), kynurenine formamidase, kynureninase, L-kynurenine hydrolase, kynurenine-3-monooxygenase, kynurenine 3-hydroxylase, 3-hydroxyanthranilate oxygenase, 3-hydroxyanthranilate 3,4-dioxygenase, kynurenine aminotransferase, and / or quinolinate phosphoribosyltransferase.

[0295] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator regulates or controls, e.g., increases or decreases, the level, availability, concentration, action, activity, metabolism, synthesis, and / or degradation of one or more metabolites in the kynurenine pathway. In some embodiments, the kynurenine pathway metabolite is L-tryptophan, n-formylkynurenine, DL-kynurenine, L-kynurenine, 3-hydroxy-DL-kynurenine, kynurenic acid, quinaldic acid, kynuramine, 3-hydroxy-L-kynurenine, 3-hydroxy-D-kynurenine, 3-hydroxyanthranilic acid, xanthommatine, anthranilic acid, xanthurenic acid, picolinic acid, quinolinic acid, and / or cinnabarinic acid, and all substantially homologous analogs and variants.

[0296] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator modulates, e.g., regulates, inhibits, or activates, the catabolism of tryptophan and / or the synthesis of N-formyl-L-kynurenine, kynurenine, and / or its derivatives, such as 3-hydroxykynurenine, kynurenic acid, and 3-hydroxyanthranilic acid, which contribute to an immunosuppressive state in the TME. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator causes, mediates, or results in an increase in the level of tryptophan (TRP) and / or a decrease in the level of kynurenine (KYN). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of IDO2 and / or tryptophan 2,3-dioxygenase (TDO).

[0297] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator modulates, e.g., regulates, inhibits, or activates, upstream and / or downstream factors or effectors in tryptophan catabolism, e.g., enzymes, e.g., components of tryptophan signaling, transcription, and / or transport and / or tryptophan catabolism or catabolic component(s), by modulating the kynurenine pathway. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator modulates, e.g., regulates, inhibits, or activates, factors or effectors involved in tryptophan catabolism, e.g., GCN2 (general control non-derepressible 2), inflammatory cues such as interferon gamma (IFNγ) and tumor necrosis factor alpha (TNFα), signal transducer and activator of transcription 3 (STAT3), mammalian target of rapamycin (mTOR), aryl hydrocarbon receptor (AHR), AHR nuclear translocator (ARNT), dioxin response element (DRE), transporters of amino acids such as TRP (tryptophan) transporters (e.g., CD98 and / or associated chains, e.g., CD98 / LAT-1 transporter complex, e.g., CD98 heavy chain (4F2hc; SLC3A2) and / or L-type amino acid transporter 1 (LAT1; SLC7A5)). , LAT2 (SLC7A8), proton-assisted amino acid transporter 4 (PAT4; SLC36A4), Asc-1, Asc-2, ATB0, B0AT1, TAT1), PR domain zinc finger protein 1 (PRDM1, also known as BLIMP-1), eukaryotic translation initiation factor 2α (eIF2α), activating transcription factor 4 (ATF4), CCAAT / enhancer binding protein homologous protein (CHOP), Gadd45α, Herp, 4E-BP1, eIF4G, JAK1, NFκB-2, FK506-BP8, and / or IFNγ-R2.

[0298] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a small molecule, a peptide, a polypeptide, an antibody or antigen-binding fragment thereof, an antibody mimetic, an aptamer or a nucleic acid molecule (e.g., siRNA), a lipid, a polysaccharide, or any combination thereof. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor or activator of a specific factor and / or enzyme in the kynurenine pathway. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an agonist or antagonist of a specific factor and / or enzyme in the kynurenine pathway. In some embodiments, the kynurenine inhibitor is an analog or derivative of one or more metabolites in the kynurenine pathway. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor that is a small molecule. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a protein or polypeptide.

[0299] The agent, e.g., a metabolic pathway modulator, e.g., a tryptophan metabolic pathway modulator, e.g., an alternative tryptophan pathway modulator, e.g., a kynurenine pathway modulator, is administered by any of a number of suitable means, for example, oral administration, injection, bolus injection, parenteral administration, intrapulmonary administration, intranasal administration, and / or intralesional administration. In some embodiments, the agent is administered locally and / or systemically. In some embodiments, the agent is added to or used to treat one or more other therapeutic agents or products, or precursors thereof. In one example, the agent is added during the process of generating cells for administration in adoptive cell therapy, e.g., cells expressing one or more engineered immune receptors such as CARs, e.g., CAR-expressing T cells.

[0300] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is tryptophan, eg, supplemental tryptophan.

[0301] IDO1 inhibitor In some embodiments, the inhibitor in the combination therapy is an inhibitor of IDO1. IDO1 belongs to a family of three heme-dependent dioxygenases, including IDO1, IDO2, and TDO, that generally play a role in catalyzing the first, rate-limiting step in the metabolism of L-tryptophan in the kynurenine pathway. The three enzymes convert L-tryptophan to N-formyl-L-kynurenine, which is then metabolized through a series of steps to form nicotinamide adenine dinucleotide (NAD).

[0302] In some embodiments, the IDO1 inhibitor is any of those described and / or summarized in the following: WO 2011 / 056652; WO 2012 / 142237; WO 2014 / 159248; WO 2008 / 058178; WO 2010 / 005958; WO 2014 / 150646; WO 2014 / 150677; WO 2015 / 002918; WO 2015 / 006520; WO 2016 / 024233; WO 2016 / 026772; WO 2015 / 082499; WO 2016 / 01 2615; 2011 / 045341; 2015 / 119944; 2006 / 005185; 2016 / 051181; 2014 / 186035; 2008 / 052352; 2009 / 073620; U.S. Patent Application Publication Nos. 2005 / 186289; 2007 / 0105907; 2016 / 0060266; 2016 / 0046596; U.S. Patent No. 7,098,209; Chinese Patent Application Publication Nos. 105567690; 103070868; 102579452; Zadori et al.,Expert Opinion on Therapeutic Patents(2016),DOI:10.1080 / 13543776.2016.1189531;Rohrig et al.,J.Med.Chem.2012,55:5270-5290;Rohrig et al. al.,J.Med.Chem.2015,58:9421-9437;Meininger et al.,Biochim.Biophys.Acta Proteins Proteomics 2011,1814:1947-1954;Tojo et al.,ACS Med.Chem.Lett.2014,5:1119-1123;Vacchelli et al. al.,(2014)OncoImmunology 3:10,e957994;Moon et al.,Journal forImmunoTherapy of Cancer(2015)3:51;Platten et al.,Front Immunol.2015 Jan 12;5:673;Weinmann, H.,ChemMedChem 2016,11:450-466;Liu et al.,Blood.2010 Apr 29;115(17):3520-30;Ninomiya et al.,Blood.2015 Jun 18;125(25):3905-16;Sheridan,C.Nature Biotechnology 33,321-322(2015);Carvalho et al. al.,Org.Biomol.Chem.2014,12:2663-2674;Eguchi et al.,Arch.Biochem.Biophys.1984,232:602-609;Peterson et al.,Med.Chem.Res.1993,3:473-482;Sono et al.,Biochemistry 1989,28:5392-5399;Opitz et al.,PLoS One. 2011;6(5):e19823; and Saito et al., Biochem. J. 1993, 291:11-14. Exemplary IDO1 inhibitors include epacadostat (INCB024360), indoximod (1-methyl-D-tryptophan), IDO peptide vaccine, and NLG919.

[0303] In some embodiments, IDO1 inhibitors include those that can increase the anti-tumor activity of various chemotherapeutic agents (e.g., platinum compounds, taxane derivatives, cyclophosphamide) without increasing toxicity.

[0304] In some embodiments, the IDO1 inhibitor is a small molecule, peptide, protein, antibody or antigen-binding fragment thereof, antibody mimetic, aptamer, or nucleic acid molecule (eg, siRNA).

[0305] In some embodiments, the IDO1 inhibitor is a small molecule. In some embodiments, the IDO1 inhibitor is a hydroxyamidine derivative, a tryptophan analog or derivative, an indole derivative or analog, a compound with a quinone or iminoquinone backbone, an imidazole derivative, such as phenylimidazole (PIM) or a phenylimidazole derivative, a 1,2,3-triazole derivative, a compound based on a thiazolotriazole backbone, an imidazothiazole derivative, a 2,3-diamino-furo[2,3-c]pyridine and a 2,3-diaminobenzo[b]thiophene derivative, a natural product inhibitor or derivative, or a single aromatic ring compound or derivative.

[0306] In some embodiments, the IDO1 inhibitor is a hydroxyamidine derivative.

[0307] In some embodiments, the IDO1 inhibitor is represented by Formula I TIFF0007623784000001.tif34128 or a pharmaceutically acceptable salt thereof, X is TIFF0007623784000002.tif17128; R 1 is Cl, Br, CF3 or CN; R 2 is H or F; and R 3 is Cl or Br.

[0308] In some embodiments, the IDO1 inhibitor is represented by Formula Ia TIFF0007623784000003.tif34128 or a pharmaceutically acceptable salt thereof.

[0309] In some embodiments, the IDO inhibitor is represented by formula Ib TIFF0007623784000004.tif37128 or a pharmaceutically acceptable salt thereof.

[0310] In some embodiments, the IDO1 inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (also known as epacadostat, INCB024360) or a pharmaceutically acceptable salt or derivative thereof. In some embodiments, the IDO1 inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-chloro-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide or a pharmaceutically acceptable salt thereof. In some embodiments, the IDO1 inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[4-fluoro-3-(trifluoromethyl)phenyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide or a pharmaceutically acceptable salt thereof. In some embodiments, the IDO1 inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N'-hydroxy-N-[3-(trifluoromethyl)phenyl]-1,2,5-oxadiazole-3-carboximidamide or a pharmaceutically acceptable salt thereof. In some embodiments, the IDO1 inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-cyano-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide or a pharmaceutically acceptable salt thereof. In some embodiments, the IDO1 inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[(4-bromo-2-furyl)methyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide or a pharmaceutically acceptable salt thereof. In some embodiments, the IDO1 inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-[(4-chloro-2-furyl)methyl]-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide or a pharmaceutically acceptable salt thereof.In some embodiments, the IDO1 inhibitor is any of the inhibitors described in WO 2008 / 058178, WO 2010 / 005958, and WO 2015 / 119944.

[0311] In some embodiments, the IDO1 inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (also known as epacadostat, INCB024360) or a pharmaceutically acceptable salt or derivative thereof.

[0312] In some embodiments, the IDO inhibitor is the following compound: TIFF0007623784000005.tif42128 or a pharmaceutically acceptable salt thereof.

[0313] In some embodiments, the IDO1 inhibitor is a tryptophan analog or derivative, or an indole derivative or analog. In some embodiments, the IDO1 inhibitor is 1-methyltryptophan (1-MT), such as 1-methyl-D-tryptophan or 1-methyl-L-tryptophan, 2,5-dihydro-L-phenylalanine, or methylthiohydantoin-D,L-tryptophan (MTH-trp or necrostatin 1). In some embodiments, the IDO1 inhibitor is brassinin and derivatives, such as S-allyl-brassinin, S-benzyl-brassinin, 5-bromo-brassinin, brassinin-derived dithiocarbamate, 1-methyltryptophan tirapazamine compound, tryptoline derivative, or tryptamine derivative. In some embodiments, the IDO1 inhibitor is 1-methyltryptophan, a competitive inhibitor of IDO1 (and IDO2) that exists as a mixture of chiral isoforms (i.e., 1-methyl-D-tryptophan and 1-methyl-L-tryptophan). In some embodiments, the IDO1 inhibitor is 1-methyl-D-tryptophan (1-D-MT; also known as indoximod or NLG8189). In some embodiments, the IDO1 inhibitor is such as any of those described in U.S. Patent Application Publication No. 2005 / 186289 and U.S. Patent No. 7,098,209. In some embodiments, the IDO1 inhibitor is 1-methyl-L-tryptophan (1-L-MT).

[0314] In some embodiments, the IDO1 inhibitor is a compound having a quinone or iminoquinone skeleton. In some embodiments, the IDO1 inhibitor is catechol, hydroquinone, p-quinone, L-dihydroxyphenylalanine, L-epinephrine, menadione, benzofuranoquinone, β-lapachone or naphthoquinone-based compounds, exiguamine A, annurine B, pyranonaphthoquinone, indolequinone, pyranonaphthoquinone, pyrroloiminoquinone, β-lapachone, cinnabarinic acid, benzofuranquinone, xestosaprol O analog, 4-iminonaphthalen-1-one derivative, NSC111041, mitomycin C or a derivative thereof. In some embodiments, the IDO1 inhibitor is any of those described in WO 2008 / 052352, WO 2006 / 005185; and Carvalho et al., Org. Biomol. Chem. 2014, 12:2663-2674, or a derivative thereof.

[0315] In some embodiments, the IDO1 inhibitor is an imidazole derivative such as phenylimidazole (PIM) or a phenylimidazole derivative. In some embodiments, the IDO1 inhibitor is 2-hydroxyphenylimidazole or 4-phenylimidazole, or a derivative thereof, such as those described in International Publication No. 2011 / 056652. In some embodiments, the IDO1 inhibitor is a PIM-based tricyclic compound, such as those described in International Publication No. 2012 / 142237 and International Publication No. 2014 / 159248. In some embodiments, the IDO1 inhibitor is 1-cyclohexyl-2-(5H-imidazo[5,1-a]isoindol-5-yl)ethanol (NLG919; also known as GDC-0919 or GTPL9019), econazole, or a derivative thereof.

[0316] In some embodiments, the IDO inhibitor is a 1,2,3-triazole derivative, for example a derivative of 4-phenyl-1,2,3-triazole, such as those described in Rohrig et al., J. Med. Chem. 2012, 55:5270-5290.

[0317] In some embodiments, the IDO1 inhibitor is a compound based on a thiazolotriazole skeleton, such as those described in Meininger et al., Biochim. Biophys. Acta Proteins Proteomics 2011, 1814:1947-1954. In some embodiments, the IDO1 inhibitor is N-(1,3-benzodioxol-5-yl)-2-{[5-(4-methylphenyl)[1,3]thiazolo[2,3-c][1,2,4]triazol-3-yl]sulfanyl}acetamide (AMG-1), or a derivative thereof.

[0318] In some embodiments, the IDO inhibitor is an imidazothiazole derivative, such as those described in Tojo et al., ACS Med. Chem. Lett. 2014, 5:1119-1123. In some embodiments, the IDO inhibitor is miconazole. In some embodiments, the IDO1 inhibitor is a compound based on a 2-aminophenylurea skeleton, such as those described in WO 2014 / 150646, WO 2014 / 150677, WO 2015 / 002918 and WO 2015 / 006520.

[0319] In some embodiments, the IDO1 inhibitor is a natural product inhibitor or a derivative thereof. In some embodiments, the IDO1 inhibitor is norharman / β-carboline, benzomalvin, halicuronic acid, thielabine, tryptanthrin, or galanal, such as those described in Eguchi et al., Arch. Biochem. Biophys. 1984, 232:602-609; Peterson et al., Med. Chem. Res. 1993, 3:473-482; Sono et al., Biochemistry 1989, 28:5392-5399; and Saito et al., Biochem. J. 1993, 291:11-14.

[0320] In some embodiments, the IDO1 inhibitor is a single aromatic ring compound or a derivative thereof, such as O-benzylhydroxylamine, phenylhydrazine, benzyl mercaptan, S-benzylisothiourea derivative, thiosemicarbazide derivative, dithiocarbamate derivative, or AC12308, or a derivative thereof, and any of those described in WO 2009 / 073620.

[0321] In some embodiments, the IDO1 inhibitor is selected from the group consisting of 2,3-diamino-furo[2,3-c]pyridine and 2,3-diaminobenzo[b]thiophene derivatives, such as (N 3 -(3-chloro-4-fluorophenyl)furo[2,3-c]pyridine-2,3-diamine or N 3 -(3-chloro-4-fluorophenyl)benzo[b]thiophene-2,3-diamine, as well as any of those described in WO 2014 / 186035.

[0322] In some embodiments, the IDO1 inhibitor is an agent that can reduce, delete, remove, knock out, knock down, or disrupt the expression of the IDO1 gene. In some embodiments, the IDO1 inhibitor is a negative regulator of the transcription factor that controls IDO1 expression. In some embodiments, the IDO1 inhibitor is a negative regulator of Bin1 or KIT signaling. In some embodiments, the inhibitor is an inhibitory nucleic acid that can reduce or control the expression of endogenous IDO1 gene. In some embodiments, the inhibitory nucleic acid is, comprises, or encodes a small interfering RNA (siRNA), a microRNA-matched shRNA, a short hairpin RNA (shRNA), a hairpin siRNA, a microRNA precursor (pre-miRNA or pri-miRNA), a microRNA (miRNA), an antisense RNA, or a ribozyme, for example, see the inhibitory nucleic acid described in Chinese Patent Publication No. 105567690, or a variant thereof, or those described in Opitz et al., PLoS One. 2011; 6(5): e19823.

[0323] In some embodiments, the IDO1 inhibitor is 1-methyl-D-tryptophan (also known as indoximod and NLG8189), 1-methyl-L-tryptophan, 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (epacadostat; INCB024360), methylthiohydantoin-D,L-tryptophan, 4-phenylimidazole, NSC401366, NLG919, F001287, PF-06840003, INCB023843, 2-hydroxyphenylimidazole, 4-phenyl-1,2,3-triazole, hydroxyamidine, a compound having a thiazolotriazole skeleton, imidazothiazole, a compound having a 2-aminophenylurea skeleton, an IDO1-targeting vaccine, and an IDO-1-targeting inhibitory nucleic acid.

[0324] In some embodiments, the IDO1 inhibitor is MTH-Trp, β-carboline, naphthoquinone compounds, S-allyl-brassinin, S-benzyl-brassinin, 5-bromo-brassinin, phenylimidazole compounds, 4-phenylimidazole, exiguamine A or NSC401366, or any of those described in Moon et al., Journal for ImmunoTherapy of Cancer (2015) 3:51.

[0325] In some embodiments, the IDO1 inhibitor includes those in clinical development for monotherapy or in combination with other anticancer agents, such as nab-paclitaxel, gemcitabine, docetaxel, ipilimumab, monoclonal antibodies targeting PDCD1, dendritic cell-based p53 vaccines, and multipeptide-based vaccines. In some embodiments, the IDO1 inhibitor is selected from among IDO1 inhibitors in clinical development, such as epacadostat (INCB024360; Developer: Incyte), indoximod (1-D-MT; Developer: NewLink Genetics), IDO peptide vaccine (Developer: Copenhagen University), F001287, PF-06840003 (Developer: Pfizer / iTeos), and NLG919 (Developer: NewLink Genetics). Exemplary IDO1 inhibitors under development for clinical trials, types of treatment and their indications include those described in Moon et al., Journal for ImmunoTherapy of Cancer (2015) 3:51.

[0326] b. Other tryptophan metabolism and / or kynurenine pathway modulators In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of IDO2 or TDO, which can also catalyze the first rate-limiting step of the kynurenine pathway. In some embodiments, the IDO1 inhibitor is a broad-spectrum inhibitor that can also inhibit IDO2 and / or TDO. In some embodiments, the inhibitor is specific for IDO1, IDO2, or TDO. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a dual-acting IDO / TDO inhibitor.

[0327] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a TDO inhibitor, such as fluorinated indole derivatives 680C91 or LM10, and / or any of those described in Dolusic et al., J Med Chem. 2011 Aug 11;54(15):5320-34. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a specific inhibitor of IDO2, such as tenatoprazole, and / or any of those described in Bakmiwewa et al., Bioorg Med Chem Lett. 2012;22(24):7641-7646. In some embodiments, the IDO1 inhibitor can inhibit both IDO1 and IDO2, such as indoximod (1-D-MT). In some embodiments, the IDO1 inhibitor can also inhibit IDO2 and / or TDO.

[0328] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator modulates a subsequent step in the kynurenine pathway, such as a reaction catalyzed by kynurenine aminotransferase, kynurenine 3-monooxygenase, kynureninase, and / or 3-hydroxyanthranilic acid 3,4-dioxygenase. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor of kynurenine aminotransferase, kynurenine 3-monooxygenase, kynureninase, and / or 3-hydroxyanthranilic acid 3,4-dioxygenase. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is selected from the group consisting of EP 2331095; EP 2420494; EP 2736337; EP 2750677; EP 2751086; EP 2833879; U.S. Patent Application Publication Nos. 2012 / 0046324; 2012 / 0329812; 2013 / 0029988; 2013 / 0331370; 2014 / 0329795; 2014 / 032981 No. 6; No. 2015 / 0057238; U.S. Patent No. 8,710,237; WO 2008 / 022286; WO 2010 / 017132; WO 2010 / 017179; WO 2011 / 091153; WO 2013 / 016488; WO 2013 / 033068; WO 2013 / 033085; WO 2013 / 033085; WO 2013 / 151707; WO 2015 / 047978 and Zadori et al., Expert Opinion on Therapeutic Patents (2016), DOI: 10.1080 / 13543776.2016.1189531.

[0329] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator regulates or controls the level, availability, concentration, action, activity, metabolism, synthesis and / or degradation of 3-hydroxyanthranilic acid (3-HAA), L-kynurenine, quinolinic acid and / or cinnabarinic acid.

[0330] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator prevents or reduces the synthesis or accumulation of kynurenine, 3-hydroxykynurenine, or 3-hydroxyanthranilic acid (3-HAA). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an inhibitor or antagonist of kynureninase. Kynureninase catalyzes the conversion of 3-hydroxykynurenine to 3-hydroxyanthranilic acid (3-HAA) and kynurenine to anthranilic acid. Inhibition of kynureninase reduces the level of 3-HAA. Exemplary kynureninase inhibitors include O-methoxybenzoylalanine 2-amino-4-[3'-hydroxyphenyl]-4-hydroxybutanoic acid and those described in U.S. Patent Application Publication No. 2015 / 0175712.

[0331] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is an enzyme, such as an enzyme involved in the kynurenine pathway, or a derivative or modified form thereof. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a kynureninase enzyme, which reduces kynurenine levels by catalyzing the catabolism of L-kynurenine and 3-hydroxy-L-kynurenine. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a modified kynureninase, such as those described in U.S. Patent Application Publication No. 2015 / 0064154.

[0332] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a modulator of other components or upstream or downstream effectors of the kynurenine pathway, e.g., components involved in tryptophan metabolism and / or signal transduction, e.g., GCN2 (general control non-derepressible 2), inflammatory cues such as interferon gamma (IFNγ), signal transducer and activator of transcription 3 (STAT3), aryl hydrocarbon receptor (AHR), AHR nuclear translocator (ARNT), dioxin response element (DRE), transporters of amino acids such as TRP (tryptophan) transporters (e.g., CD98 and / or associated chains, e.g., CD98 / LAT-1 transporter complex, e.g., CD98 heavy chain (4F2hc; SLC3A2) and / or L-type amino acid transporter 1 (LAT1; SLC7A5), LAT2 (SLC7A8), proton-assisted amino acid transporter (PAT1; SLC7A8), and the like. and one or more of paraoxygenase 4 (PAT4; SLC36A4), Asc-1, Asc-2, ATB0, B0AT1, TAT1), PR domain zinc finger protein 1 (PRDM1, also known as BLIMP-1), eukaryotic translation initiation factor 2α (eIF2α), activating transcription factor 4 (ATF4), CCAAT / enhancer binding protein homologous protein (CHOP), Gadd45α, Herp, 4E-BP1, eIF4G, JAK1, NFκB-2, FK506-BP8, and / or IFNγ-R2 (e.g., McGaha et al. (2012) Immunol. Rev. 249(1):135-157; Munn et al. (2005) Immunity 22; 633-642; see Broer et al. (2011) Biochem. J. 436: 193-211).

[0333] 2. Compositions and Formulations In some embodiments of the methods, compositions, combinations, kits and uses provided herein, the combination therapy can be administered with one or more compositions, e.g., pharmaceutical compositions comprising a tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, and / or an immunotherapy, e.g., a T cell therapy.

[0334] In some embodiments, compositions, e.g., pharmaceutical compositions comprising a tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, can include a carrier, such as a diluent, adjuvant, excipient, or vehicle, administered with the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, and / or cells. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E.W. Martin. Such compositions generally contain a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, in purified form, together with a suitable amount of carrier to provide the form for proper administration to a patient. Such pharmaceutical carriers can be sterile liquids, e.g., water, and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, and sesame oil. Saline solutions and aqueous dextrose and glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. The pharmaceutical composition can include any one or more of diluents, adjuvants, anti-adherents, binders, coating agents, fillers, flavorings, coloring agents, lubricants, glidants, preservatives, surfactants, adsorbents, emulsifiers, pharmaceutical excipients, pH buffers, or sweeteners, and combinations thereof. In some embodiments, the pharmaceutical composition can be in liquid, solid, lyophilized powder, gel form, and / or combinations thereof. In some embodiments, the choice of carrier is determined in part by the particular inhibitor and / or method of administration.

[0335] Pharmaceutically acceptable carriers are generally non-toxic to recipients at the dosages and concentrations employed, and include, but are not limited to, buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol); (less than 100%) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants, stabilizers, and / or preservatives such as polyethylene glycol (PEG). Compositions containing tryptophan metabolism and / or kynurenine pathway modulators, for example, IDO1 inhibitors, can also be lyophilized.

[0336] In some embodiments, the pharmaceutical composition can be formulated for administration by any route known to those skilled in the art, including intramuscular, intravenous, intradermal, intralesional, intraperitoneal injection, subcutaneous, intratumoral, epidural, nasal, oral, vaginal, rectal, topical, local, otic, inhalation, buccal (e.g., sublingual), and transdermal administration, or any route. In some embodiments, other modes of administration are also contemplated. In some embodiments, administration is by bolus injection, injection, such as intravenous or subcutaneous injection, intraocular injection, periocular injection, subretinal injection, intravitreal injection, transseptal injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjunctival injection, subconjunctival injection, sub-Tenon injection, retrobulbar injection, peribulbar injection, or posterior parascleral delivery. In some embodiments, administration is by parenteral administration, intrapulmonary administration, and intranasal administration, and, if desired for local treatment, by intralesional administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In some embodiments, a given dose is administered by a single bolus injection. In some embodiments, it is administered by multiple bolus injections over a period of, for example, 3 days or less, or by continuous infusion.

[0337] In some embodiments, administration can be topical, local, or systemic depending on the location of treatment. In some embodiments, local administration to the area requiring treatment can be achieved, for example, but not limited to, by local infusion during surgery, by topical application in combination with a wound dressing after surgery, by injection, by using a catheter, by using a suppository, or by using an implant. In some embodiments, the composition can also be administered together with other biologically active agents, continuously, intermittently, or in the same composition. In some embodiments, administration can also include controlled release systems, including controlled release formulations and device-controlled release by pumps, etc. In some embodiments, administration is oral administration.

[0338] In some embodiments, pharmaceutically and therapeutically active compounds and their derivatives are typically formulated and administered in unit or multiple dosage forms. Each unit dosage contains a predetermined amount of the therapeutically active compound sufficient to produce the desired therapeutic effect, along with the necessary pharmaceutical carrier, vehicle, or diluent. In some embodiments, unit dosage forms include, but are not limited to, tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, oral solutions or suspensions, and oil-and-water emulsions, each containing an appropriate amount of the compound or a pharmaceutically acceptable derivative thereof. Unit dosage forms may be sealed ampoules and syringes, or individually packaged tablets or capsules. Unit dosage forms may be administered in fractions or multiples thereof. In some embodiments, multiple dosage forms are multiple identical unit dosage forms packaged in a single container to be administered in separate unit dosage forms. Examples of multiple dosage forms include vials, bottles of tablets or capsules, or bottles of pints or gallons.

[0339] 3. Dosage and Schedule of Tryptophan Metabolism and / or Kynurenine Pathway Modulators In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a tryptophan metabolism and / or kynurenine pathway modulator, e.g., an inhibitor of IDO1, and an immunotherapy, e.g., T cell therapy (e.g., CAR-expressing T cells) or T cell-associated therapy. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an inhibitor of IDO1, is administered prior to, after, during, during the course of, simultaneously with, about simultaneously with, continuously, and / or intermittently with the administration of the immunotherapy, e.g., T cell therapy (e.g., CAR-expressing T cells) or T cell-associated therapy. In some embodiments, the method comprises administering the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an inhibitor of IDO1, prior to administration of the T cell therapy. In other embodiments, the method comprises administering the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an inhibitor of IDO1, after administration of the T cell therapy. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an inhibitor of IDO1, is not further administered after initiation of T cell therapy. In some embodiments, the administration schedule comprises administering the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an inhibitor of IDO1, before and after initiation of T cell therapy. In some embodiments, the administration schedule comprises administering the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an inhibitor of IDO1, concurrently with administration of T cell therapy.

[0340] In some embodiments, the tryptophan metabolism and / or kynurenine pathway regulator, for example, an IDO1 inhibitor, is administered multiple times in multiple doses. In some embodiments, the tryptophan metabolism and / or kynurenine pathway regulator, for example, an IDO1 inhibitor, is administered once. In some embodiments, the tryptophan metabolism and / or kynurenine pathway regulator, for example, an IDO1 inhibitor, is administered 6 times a day, 5 times a day, 4 times a day, 3 times a day, 2 times a day, once a day, every other day, once every 3 days, twice a week, once a week, or only once before or after the start of immunotherapy (e.g., T cell therapy, such as CAR-T cell therapy). In some aspects, the administration frequency of the tryptophan metabolism and / or kynurenine pathway regulator, for example, an IDO1 inhibitor, is twice a week, once every 14 days, once every 21 days, or once a month. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered in multiple doses at regular intervals before, during, and / or after the administration period of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered in a single dose or in multiple doses at regular intervals before the administration of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy). In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered once or in multiple doses at regular intervals after the administration of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy). In some embodiments, one or more administrations of the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, can be administered simultaneously with the administration of a dose of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy).

[0341] In some embodiments, the dose, frequency, duration, timing and / or sequence of administration of the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is determined based on certain thresholds or criteria resulting from the screening step and / or an assessment of a treatment outcome described herein, e.g., a treatment outcome described in Section III herein.

[0342] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered prior to the administration of immunotherapy (e.g., T cell therapy, such as CAR-T cell therapy). In some aspects, the administration of the tryptophan metabolism and / or kynurenine pathway modulator is timed to allow sufficient time to increase tryptophan (TRP) levels or decrease kynurenine (KYN) levels in a subject's biological sample, e.g., in the tumor microenvironment (TME) of the tumor being treated. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered at a time when the modulator (e.g., an IDO1 inhibitor) has caused or is likely to have caused a change in the level of a metabolite in the kynurenine pathway, such as an increase in the level of TRP or a decrease in the level of KYN or a KYN derivative (e.g., an increase or decrease of at least at least 1.2-fold, at least 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 10-fold, or more, or at least about 1.2-fold, at least about 1.5-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 10-fold, or more), compared to the level at the time point immediately before administration of the tryptophan metabolism and / or kynurenine pathway modulator. Exemplary methods for assessing or determining parameters associated with changes in the level of such metabolites in the kynurenine pathway are described in Section III.

[0343] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered 0 to 90 days, e.g., 0 to 30 days, 0 to 15 days, 0 to 6 days, 0 to 96 hours, 0 to 24 hours, 0 to 12 hours, 0 to 6 hours, or 0 to 2 hours, 2 hours to 30 days, 2 hours to 15 days, 2 hours to 6 days, 2 hours to 96 hours, 2 hours to 24 hours, 2 hours to 12 hours, 2 hours to 6 hours, 6 hours to 90 days, 6 hours to 30 days, 6 hours to 15 days, 6 hours to 6 days, The drug is administered 6 to 96 hours, 6 to 24 hours, 6 to 12 hours, 12 hours to 90 days, 12 hours to 30 days, 12 hours to 15 days, 12 hours to 6 days, 12 hours to 96 hours, 12 hours to 24 hours, 24 hours to 90 days, 24 hours to 30 days, 24 hours to 15 days, 24 hours to 6 days, 24 hours to 96 hours, 96 hours to 90 days, 96 hours to 30 days, 96 hours to 15 days, 96 hours to 6 days, 6 days to 90 days, 6 days to 30 days, 6 days to 15 days, 15 days to 90 days, 15 days to 30 days, or 30 days to 90 days, or approximately within the aforementioned periods. In some aspects, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered within about 96 hours, 72 hours, 48 ​​hours, 24 hours, 12 hours, 6 hours, 2 hours, or 1 hour prior to the initiation of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy).

[0344] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered at least 1 hour or at least about 1 hour, at least 2 hours or at least about 2 hours, at least 6 hours or at least about 6 hours, at least 12 hours or at least about 12 hours, at least 1 day or at least about 1 day, at least 2 days or at least about 2 days, at least 3 days or at least about 3 days, at least 4 days or at least about 4 days, at least 5 days or at least about 5 days, at least 6 days or at least about 1 hour prior to the start of administration of the immunotherapy (e.g., T cell therapy such as CAR-T cell therapy). The administration is at least about 6 days, at least about 7 days, at least about 12 days, at least about 14 days, at least about 15 days, at least about 21 days, at least about 24 days, at least about 28 days, at least about 30 days, at least about 35 days, at least about 42 days, at least about 60 days, or at least about 60 days, or at least about 90 days. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered up to 2, 3, 4, 5, 6, 7, 8, 12, 14, 15, 21, 24, 28, 30, 35, 42, 60, or 90 days prior to the start of administration of the immunotherapy (e.g., T cell therapy, such as CAR-T cell therapy). In some embodiments, administration of the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is initiated more than 1 day prior to the start of administration of the immunotherapy (e.g., T cell therapy, such as CAR-T cell therapy).

[0345] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered after administration (e.g., initiation of administration) of a T cell therapy, such as CAR-T cell therapy, but before the time point at which a portion of T cells reach peak or maximal proliferation, prolonged tryptophan starvation, exhaustion, anergy, cell death, regression, suppressed anti-tumor immune response, or tolerance to tumor antigens, and / or are unable to recover from and / or reverse the effects of starvation, e.g., tryptophan starvation. In some cases, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered before the time point at which T cells exhibit slowed or halted proliferation and / or functional activity due to tryptophan depletion. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered less than or about 1 hour, less than or about 2 hours, less than or about 6 hours, less than or about 12 hours, less than or about 1 day, less than or about 2 days, less than or about 3 days, less than or about 4 days, less than or about 5 days, less than or about 6 days, less than or about 7 days after administration (e.g., initiation of administration) of T cell therapy. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered within 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 5 days, 6 days, 7 days, 14 days, 21 days, 28 days, or within about said time periods after initiation of T cell therapy. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered within, or about, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 5 days, 6 days, or 7 days after initiation of T cell therapy. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered within, or about, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, or 14 days after initiation of T cell therapy.In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered within or about 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, or 48 hours after initiation of T cell therapy.

[0346] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered after administration (e.g., initiation of administration) of T cell therapy, such as CAR-T cell therapy, at or after the subject receiving T cell therapy exhibits increased IDO1 expression and / or expression of tryptophan transporters, e.g., amino acid transporters such as LAT1, LAT2, CD98hc, and / or PAT4, in a tumor sample from the subject compared to a sample obtained before administration of T cell therapy, a baseline, and / or a sample from a different subject. In some embodiments, the subject is identified or selected for administration of a tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, if a tumor sample from the subject exhibits increased IDO1 expression and / or expression of tryptophan transporters, e.g., amino acid transporters such as LAT1, LAT2, CD98hc, and / or PAT4, compared to a sample obtained before administration of T cell therapy, a baseline, and / or a sample from a different subject.

[0347] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered at a time when there is a decrease in tryptophan levels, an increase in kynurenine levels, and / or an increase in IDO1, IDO2, or TDO expression or activity in a biological sample from the subject compared to before administration of T cell therapy or compared to an earlier time point after administration of T cell therapy; before the number of T cell therapy cells detectable in the blood from the subject is decreased compared to in the subject at an earlier time point after administration of T cell therapy; before the T cells exhibit prolonged tryptophan starvation, depletion, decreased effector function, expression of inhibitory receptors, and / or loss of proliferative capacity; before peak or maximum levels of T cell therapy cells are detectable in the blood of the subject; and / or when levels of interferon-gamma (IFNγ) are increased in a biological sample from the subject compared to before administration of T cell therapy or compared to an earlier time point after administration of T cell therapy. In some embodiments, the increase or decrease is greater than 1.2-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, or more, or greater than about 1.2-fold, about 1.5-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 10-fold, or more.

[0348] In some of any such embodiments in which tryptophan metabolism and / or kynurenine pathway modulation is given prior to immunotherapy (e.g., T cell therapy, such as CAR-T cell therapy), administration of the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is continued at regular intervals until and / or for a period after initiation of immunotherapy.

[0349] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered after administration of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy). In some embodiments, the administration of the tryptophan metabolism and / or kynurenine pathway modulator is at a time when the prior administration of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy) has directly or indirectly caused or induced, or is likely to have caused or induced, a change in the level of a metabolite in the kynurenine pathway, such as a decrease in the level of TRP or an increase in the level of KYN or a KYN derivative (e.g., at least a 1.2-fold, at least a 1.5-fold, at least a 2-fold, at least a 3-fold, at least a 4-fold, at least a 5-fold, at least a 6-fold, at least a 7-fold, at least a 8-fold, at least a 10-fold, or more increase or decrease, or at least about a 1.2-fold, at least about a 1.5-fold, at least a 2-fold, at least a 3-fold, at least a 4-fold, at least a 5-fold, at least a 6-fold, at least a 7-fold, at least a 8-fold, at least a 10-fold, or more increase or decrease), compared to the level at a time immediately prior to or after the initiation of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy). Exemplary methods for assessing or determining parameters associated with changes in the levels of such metabolites in the kynurenine pathway are described in Section III.

[0350] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is administered within, or about, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 96 hours, 4 days, 5 days, 6 days, 7 days, 14 days, 15 days, 21 days, 24 days, 28 days, 30 days, 36 days, 42 days, 60 days, 72 days, or 90 days after initiation of administration of immunotherapy (e.g., T cell therapy). In some embodiments, provided methods comprise continued administration of the tryptophan metabolism and / or kynurenine pathway modulator, such as at regular intervals, after initiation of administration of immunotherapy.

[0351] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered for up to or about 1 day, up to or about 2 days, up to or about 3 days, up to or about 4 days, up to or about 5 days, up to or about 6 days, up to or about 7 days, up to or about 12 days, up to or about 14 days, up to or about 14 days, or up to 21 days after administration of an immunotherapy (e.g., a T cell therapy such as CAR-T cell therapy). or for up to or about 21 days, up to or about 24 days, up to or about 28 days, up to or about 30 days, up to or about 35 days, up to or about 42 days, up to or about 60 days, up to or about 90 days, up to or about 120 days, up to or about 180 days, up to or about 240 days, or up to or about 360 days, or more.

[0352] In some of any such above embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered before and after initiation of immunotherapy (e.g., T cell therapy such as CAR-T cell therapy).

[0353] In some embodiments of the methods provided herein, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, and the immunotherapy (e.g., T cell therapy such as CAR-T cell therapy) are administered simultaneously or near simultaneously.

[0354] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator (e.g., an IDO1 inhibitor) is administered twice daily.

[0355] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered in a dose of about 0.02 mg / kg to about 200 mg / kg of the subject's body weight (mg / kg), e.g., 0.02 mg / kg to 100 mg / kg, 0.02 mg / kg to 100 mg / kg, 0.02 mg / kg to 50 mg / kg, 0.02 mg / kg to 10 mg / kg, 0.02 mg / kg to 1.0 mg / kg, 0.02 mg / kg to 0.2 mg / kg, 0.2 mg / kg to 100 mg / kg, 0.2 mg / kg to 50 mg / kg, They are independently administered at a dose of 0.2 mg / kg to 10 mg / kg, 0.2 mg / kg to 1.0 mg / kg, 1.0 mg / kg to 200 mg / kg, 1.0 mg / kg to 100 mg / kg, 1.0 mg / kg to 50 mg / kg, 1.0 mg / kg to 10 mg / kg, 10 mg / kg to 200 mg / kg, 10 mg / kg to 100 mg / kg, 10 mg / kg to 50 mg / kg, 50 mg / kg to 200 mg / kg, 50 mg / kg to 100 mg / kg, or 100 mg / kg to 200 mg / kg, or approximately within the above ranges.In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered at 0.02 mg / kg, 0.03 mg / kg, 0.04 mg / kg, 0.05 mg / kg, 0.06 mg / kg, 0.07 mg / kg, 0.08 mg / kg, 0.09 mg / kg, 0.1 mg / kg, 0.15 mg / kg, 0.2 mg / kg, 0.25 mg / kg, 0.1 ... .30mg / kg, 0.35mg / kg, 0.40mg / kg, 0.45mg / kg, 0.5mg / kg, 0.55mg / kg, 0.6mg / kg, 0.7mg / kg, 0.8mg / kg , 0.9mg / kg, 1.0mg / kg, 1.1mg / kg, 1.2mg / kg, 1.3mg / kg, 1.4mg / kg, 1.5mg / kg, 1.6mg / kg, 1.7mg / kg, 1.8 1.9mg / kg, 2mg / kg, 2.5mg / kg, 3mg / kg, 3.5mg / kg, 4mg / kg, 4.5mg / kg, 5mg / kg, 5.5mg / kg, 6mg / kg, 6.5mg / kg, 7mg / kg, 7.5mg / kg, 8mg / kg, 8.5mg / kg, 9mg / kg, 9.5mg / kg, 10mg / kg, 11mg / kg, 12mg / kg, 13mg / kg, 14mg / kg, 15mg / kg, 16mg / kg, 17mg / kg, 18mg / kg, 19mg / kg, 20mg / kg, 21mg / kg, 22mg / kg, 23mg / kg, 24mg / kg, 25mg / kg, 50mg / kg, 75mg / kg, 100mg / kg, or 250mg / kg or more.

[0356] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered in a range of 2.5 mg to 5000 mg, e.g., 2.5 mg to 2000 mg, 2.5 mg to 1000 mg, 2.5 mg to 500 mg, 2.5 mg to 200 mg, 2.5 mg to 100 mg, 2.5 mg to 50 mg, 2.5 mg to 25 mg, 2.5 mg to 10 mg, 25 mg to 5000 mg, 25 mg to 2000 mg, 25 mg to 1000 mg, 25 mg to 500 mg, 25 mg to 200 mg, 25 mg to 100 mg, 25 mg to 50 mg, 50 mg to 5000 mg, 50 mg to 2000 mg, 50 mg, inclusive. 200mg-1000mg, 50mg-500mg, 50mg-200mg, 50mg-100mg, 100mg-5000mg, 100mg-2000mg, 100mg-1000mg, 100mg-500mg, 100mg-200mg, 200mg-5000mg, 200mg-2000mg, 200mg-1000mg, 200mg-500mg, 500mg-5000mg, 500mg-2000mg, 500mg-1000mg, or 1000mg-2000mg, or each dose of tryptophan metabolism and / or kynurenine pathway modulator is administered independently.

[0357] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered in a range of 2.5 mg to 5000 mg, e.g., 2.5 mg to 2000 mg, 2.5 mg to 1000 mg, 2.5 mg to 500 mg, 2.5 mg to 200 mg, 2.5 mg to 100 mg, 2.5 mg to 50 mg, 2.5 mg to 25 mg, 2.5 mg to 10 mg, 25 mg to 5000 mg, 25 mg to 2000 mg, 25 mg to 1000 mg, 25 mg to 500 mg, 25 mg to 200 mg, 25 mg to 100 mg, 25 mg to 50 mg, 50 mg to 5000 mg, 50 mg to 2000 mg, 50 mg to or 1000 mg, 50 mg to 500 mg, 50 mg to 200 mg, 50 mg to 100 mg, 100 mg to 5000 mg, 100 mg to 2000 mg, 100 mg to 1000 mg, 100 mg to 500 mg, 100 mg to 200 mg, 200 mg to 5000 mg, 200 mg to 2000 mg, 200 mg to 1000 mg, 200 mg to 500 mg, 500 mg to 5000 mg, 500 mg to 2000 mg, 500 mg to 1000 mg, or 1000 mg to 2000 mg, or each dose of tryptophan metabolism and / or kynurenine pathway modulator is administered orally.

[0358] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is orally administered six times per day, five times per day, four times per day, three times per day, twice per day, once per day, every other day, every three days, twice per week, or once per week, or each dose of the tryptophan metabolism and / or kynurenine pathway modulator is orally administered independently. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered six times per day, five times per day, four times per day, three times per day, twice per day, or once per day, or each dose of the tryptophan metabolism and / or kynurenine pathway modulator is administered independently.

[0359] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered at a dose of at least 5 mg / day, at least 10 mg / day, at least 25 mg / day, at least 50 mg / day, at least 100 mg / day, at least 200 mg / day, at least 400 mg / day, at least 500 mg / day, at least 600 mg / day, at least 800 mg / day, at least 1000 mg / day, at least 1200 mg / day, at least 1600 mg / day, at least 2000 mg / day, at least 5000 mg / day, or at least 10000 mg / day, or at least about 5 mg / day , at least about 10 mg / day, at least about 25 mg / day, at least about 50 mg / day, at least about 100 mg / day, at least about 200 mg / day, at least about 400 mg / day, at least about 500 mg / day, at least about 600 mg / day, at least about 800 mg / day, at least about 1000 mg / day, at least about 1200 mg / day, at least about 1600 mg / day, at least about 2000 mg / day, at least about 5000 mg / day, or at least about 10000 mg / day, or wherein each dose of tryptophan metabolism and / or kynurenine pathway modulator is administered independently.

[0360] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered in an amount of at least 0.02 mg / kg, at least 0.2 mg / kg, at least 1 mg / kg, at least 3 mg / kg, at least 6 mg / kg, at least 10 mg / kg, at least 20 mg / kg, at least 30 mg / kg, at least 50 mg / kg, at least 100 mg / kg, or at least 200 mg / kg, or at least At least about 0.02 mg, at least about 0.2 mg / kg, at least about 1 mg / kg, at least about 3 mg / kg, at least about 6 mg / kg, about 10 mg / kg, at least about 20 mg / kg, at least about 30 mg / kg, at least about 50 mg / kg, at least about 100 mg / kg, or at least about 200 mg / kg, or at least about 0.02 mg, at least about 0.2 mg / kg, at least about 1 mg / kg, at least about 3 mg / kg, at least about 6 mg / kg, about 10 mg / kg, about 20 mg / kg, at least about 30 mg / kg g, at a dose of about 50 mg / kg, about 100 mg / kg, or about 200 mg / kg, or at least 2.5 mg, at least 25 mg, at least 50 mg, at least 100 mg, at least 200 mg, at least 400 mg, at least 500 mg, at least 600 mg, at least 800 mg, at least 1000 mg, at least 1200 mg, at least 1600 mg, at least 2000 mg, or at least 5000 mg, or at least about 2.5 mg, at least about 25 mg, at least about 50 mg, at least about 100 mg, at least about 200 mg, at least about 400 mg, at least about 500 mg, at least about 600 mg, at least about 800 mg, at least about 1000 mg, at least about 1200 mg, at least about 1600 mg, at least about 2000 mg, or at least about 5000 mg, or each dose of tryptophan metabolism and / or kynurenine pathway modulator is administered independently.

[0361] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a protein or polypeptide or derivative thereof, and the tryptophan metabolism and / or kynurenine pathway modulator is administered at a dosage of about 1 μg / kg body weight, about 5 μg / kg body weight, about 10 μg / kg body weight, about 50 μg / kg body weight, about 100 μg / kg body weight, about 200 μg / kg body weight, about 350 μg / kg body weight, about 500 μg / kg body weight, about 1 mg / kg body weight, about 5 mg / kg body weight, about 10 mg / kg body weight, about 50 mg / kg body weight, about 100 mg / kg body weight, about 200 mg / kg body weight, about 350 mg / kg body weight, about 500 mg / kg body weight, about 1000 mg / kg body weight or more per administration, and any range therebetween.

[0362] In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor, is administered intravenously, e.g., by intravenous injection, or each dose of the tryptophan metabolism and / or kynurenine pathway modulator is administered independently. In some embodiments, the tryptophan metabolism and / or kynurenine pathway modulator is a protein or polypeptide or derivative thereof, and the tryptophan metabolism and / or kynurenine pathway modulator is administered at a dosage of about 1 μg / kg body weight, about 5 μg / kg body weight, about 10 μg / kg body weight, about 50 μg / kg body weight, about 100 μg / kg body weight, about 200 μg / kg body weight, about 350 μg / kg body weight, about 500 μg / kg body weight, about 1 mg / kg body weight, about 5 mg / kg body weight, about 10 mg / kg body weight, about 50 mg / kg body weight, about 100 mg / kg body weight, about 200 mg / kg body weight, about 350 mg / kg body weight, about 500 mg / kg body weight, about 1000 mg / kg body weight or more per administration, and any range therebetween.

[0363] C. Lymphodepletion Therapy In some embodiments, the provided methods may further include administering one or more lymphodepleting therapies, such as before or simultaneously with the administration of immunotherapy, such as T cell therapy (e.g., CAR-expressing T cells) or T cell combination therapy. In some embodiments, the lymphodepleting therapy includes the administration of a phosphamide, such as cyclophosphamide. In some embodiments, the lymphodepleting therapy may include the administration of fludarabine. In some embodiments, fludarabine is excluded from the lymphodepleting therapy. In some embodiments, no lymphodepleting therapy is administered.

[0364] In some embodiments, subjects pretreated with immunodepleting therapy (e.g., lymphodepleting therapy) can improve the efficacy of adoptive cell therapy (ACT). Pretreatment with lymphodepleting agents, including a combination of cyclosporine and fludarabine, is effective in improving the outcome of tumor-infiltrating lymphocyte (TIL) cells transferred in cell therapy, including improving the response and / or persistence of the administered cells. See, for example, Dudley et al., Science, 298, 850-54 (2002); Rosenberg et al., Clin Cancer Res, 17(13):4550-4557 (2011). Similarly, CAR + In relation to T cells, several trials have incorporated lymphodepleting agents, most commonly cyclophosphamide, fludarabine, bendamustine, or combinations thereof, sometimes accompanied by low-dose irradiation. See Han et al., Journal of Hematology & Oncology, 6:47 (2013); Kochenderfer et al., Blood, 119:2709-2720 (2012); Kalos et al., Sci Transl Med, 3(95):95ra73 (2011); Clinical Trial Registration Numbers NCT02315612; NCT01822652.

[0365] Such pretreatment may be performed with the goal of reducing the risk of one or more of a variety of consequences that may diminish the activity, outcome, or response of the treatment or to the treatment. These include the phenomenon known as "cytokine sink," in which T cells, B cells, and NK cells compete with TILs for homeostasis and activation of cytokines such as IL-2, IL-7, and / or IL-15; suppression of TILs by regulatory T cells, NK cells, or other cells of the immune system; and the influence of negative regulators in the tumor microenvironment. Muranski et al., Nat Clin Pract Oncol. December; 3(12):668-681 (2006).

[0366] In some embodiments, the provided method further comprises administering a lymphodepleting therapy to the subject. In some embodiments, the method comprises administering the lymphodepleting therapy to the subject before administering the cell dose for the T cell-based therapy. In some embodiments, the lymphodepleting therapy comprises a chemotherapeutic agent such as fludarabine and / or cyclophosphamide. In some embodiments, the lymphodepleting therapy does not comprise fludarabine. In some embodiments, the lymphodepleting therapy comprises only cyclophosphamide. In some embodiments, the administration of the cells and / or the lymphodepleting therapy is performed by outpatient delivery.

[0367] In some embodiments, the method further comprises administering to the subject a conditioning agent, such as a lymphodepleting agent or a chemotherapeutic agent, for example, cyclophosphamide, fludarabine, or a combination thereof, prior to administering the cell dose. For example, the subject may be administered the conditioning agent at least 2 days, for example, at least 3, 4, 5, 6, or 7 days, prior to the first or subsequent administration of immunotherapy, such as T cell therapy (e.g., CAR-expressing T cells) or T cell conjugate therapy. In some embodiments, the subject is administered the conditioning agent within 7 days, for example, within 6, 5, 4, 3, or 2 days prior to administering the cell dose.

[0368] In some embodiments, the subject is pretreated with cyclophosphamide at a dose of 20 mg / kg to 100 mg / kg or about 20 mg / kg to 100 mg / kg, e.g., 40 mg / kg to 80 mg / kg or about 40 mg / kg to 80 mg / kg. In some embodiments, the subject is pretreated with cyclophosphamide at 60 mg / kg or about 60 mg / kg. In some embodiments, cyclophosphamide can be administered in a single dose or in multiple doses, such as daily, every other day, or every three days. In some embodiments, cyclophosphamide is administered once daily for one or two days.

[0369] In some embodiments, when the lymphodepleting agent comprises fludarabine, the subject is administered 1 mg / m 2 ~100mg / m 2 or approximately 1 mg / m 2 ~100mg / m 2 , e.g., 10 mg / m 2 ~75mg / m 2 , 15 mg / m 2 ~50mg / m 2 , 20 mg / m 2 ~30mg / m 2 , or 24 mg / m 2 ~26mg / m 2 In some cases, the subject is administered fludarabine at a dose of 25 mg / m 2 In some embodiments, fludarabine is administered in a single dose or in multiple doses, such as every day, every other day, or every three days. In some embodiments, fludarabine is administered daily for one to five days, such as three to five days.

[0370] In some embodiments, the lymphodepleting agent comprises a combination of agents, such as a combination of cyclophosphamide and fludarabine. Thus, the combination of agents can include cyclophosphamide at any dose or administration schedule as described above, and fludarabine at any dose or administration schedule as described above. For example, in some aspects, the subject is administered 60 mg / kg (~2 g / m) of cyclophosphamide prior to the cell dose.2 ) of cyclophosphamide and 3 to 5 doses of 25 mg / m 2 of fludarabine is administered.

[0371] In some embodiments, prior to receiving T cells, e.g., CAR-expressing T cells, the subject is administered a tryptophan metabolism and / or kynurenine pathway modulator at least 1, 2, 3, 4, 5, 6, 7, 14, 21, 28, or 30 days prior to administration of the cells, and is administered lymphodepleting conditioning chemotherapy of cyclophosphamide at least 2 days prior to administration of the T cells, e.g., CAR-expressing T cells, and generally within 7 days prior to administration of the cells. In some cases, for example, cyclophosphamide is administered at least 1, 2, 3, 4, 5, 6, 7, 14, 21, 28, or 30 days after administration of the tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor. Following the conditioning treatment, the subject is administered a dose of CAR-expressing T cells as described herein.

[0372] In some embodiments, administering a conditioning agent, for example, for lymphodepletion therapy, before injecting a cell dose improves the outcome of the treatment.For example, in some aspects, the conditioning improves the efficacy, outcome or activity of the cell dose treatment in the subject, or enhances the persistence of recombinant receptor-expressing cells (e.g., CAR-expressing cells, such as CAR-expressing T cells).In some embodiments, the conditioning treatment enhances disease-free survival, such as the proportion of subjects who are alive and do not show minimal residual disease or molecularly detectable disease after a given period of time after administration of cells.In some embodiments, the time to median disease-free survival increases.

[0373] Once the cells are administered to a subject (e.g., a human), the biological activity of the engineered cell population in some aspects is measured by any of a number of known methods or any of the evaluation methods or assays described herein, e.g., in Section III. Evaluated parameters include specific binding of engineered or natural T cells or other immune cells to an antigen in vivo, e.g., by imaging, or ex vivo, e.g., by ELISA or flow cytometry. In certain embodiments, the ability of the engineered cells to destroy target cells can be measured using any suitable method known in the art, such as the cytotoxicity assays described in Kochenderfer et al., J. Immunotherapy, 32(7):689-702 (2009) and Herman et al., J. Immunological Methods, 285(1):25-40 (2004). In certain embodiments, the biological activity of the cells can also be measured by assaying the expression and / or secretion of specific cytokines. In some aspects, biological activity is measured by assessing a clinical outcome, such as a reduction in tumor burden or burden. In some aspects, the toxic outcome, persistence and / or proliferation of the cells, and / or the presence or absence of a host immune response is assessed.

[0374] In some embodiments, administration of a pretreatment agent, e.g., lymphodepletion therapy, prior to infusion of the cell dose improves the outcome of the treatment, such as by improving the efficacy, outcome, or activity of the cell dose treatment, or enhances the persistence of the recombinant receptor-expressing cells (e.g., CAR-expressing cells, such as CAR-expressing T cells) in the subject. In some embodiments, lymphodepletion therapy is not administered prior to administration of immunotherapy, e.g., T cell therapy, and / or a tryptophan metabolism and / or kynurenine pathway modulator, e.g., an IDO1 inhibitor.

[0375] III. Screening, evaluation, or selection of subjects and evaluation of treatment outcomes In some embodiments of the methods, compositions, combinations, kits, and uses provided herein, the combination therapy may further include one or more screening and / or evaluation steps to determine the suitability of the combination therapy and to identify or select subjects for treatment with the combination therapy and / or continuation of the combination therapy, and / or a step for evaluating and / or monitoring the treatment outcome. In some embodiments, the screening step may include a step of identifying subjects for the treatment, such as evaluating or assessing specific parameters such as those described herein. In some embodiments, the step for evaluating the treatment outcome may include a step for evaluating and / or monitoring the treatment and / or identifying subjects for administration of additional or remaining steps of the treatment and / or for repeat therapy. In some embodiments, the screening step and / or evaluation of the treatment outcome can be used to determine the dose, frequency, duration, timing, and / or sequence of the combination therapy provided herein.

[0376] In some embodiments, the provided combination therapy results in any one or more therapeutic outcomes, such as characteristics associated with any one or more of the parameters associated with the therapy or treatment, as described below.

[0377] In some embodiments, combination therapy can be used to overcome the immunosuppressive state of the TME and enhance the activity of immunotherapies such as T cell therapy (e.g., CAR-expressing T cells) or T cell-combined therapy. In some embodiments, modulation of the kynurenine pathway, for example, tryptophan (TRP) depletion and / or inhibition of the production of kynurenine (KYN) and its derivatives, results in enhanced T cell proliferation and / or activity, thereby increasing the activity of immunotherapies such as T cell therapy (e.g., CAR-expressing T cells) or T cell-combined therapy compared to the activity of the immunotherapy alone. In some embodiments, screening processes and / or evaluation of treatment outcomes can be used to determine and / or monitor the activity of combination therapy.

[0378] In some aspects, screening or evaluation of the specific parameters described can be used to identify subjects who are suitable, sensitive, and / or responsive to a particular treatment, such as the combination therapy provided herein, or to monitor the treatment and / or identify effective dosages for subjects or subgroups or other groups of subjects. In some embodiments, such screening or evaluation can be used to select or identify patients who are predicted to be responsive to the treatment. In some embodiments, the screening process is used to determine the dose, frequency, duration, timing, and / or sequence of the combination therapy provided herein.

[0379] In some embodiments, any of the screening steps and / or evaluations of treatment outcomes described herein can be used before, during, during the course of, or after administration of one or more steps of a provided combination therapy, e.g., immunotherapy such as T cell therapy (e.g., CAR-expressing T cells) or T cell binding therapy, and / or tryptophan metabolism and / or kynurenine pathway modulator. In some embodiments, the evaluation is performed before, during, during the course of, or after performing any of the methods provided herein. In some embodiments, the evaluation is performed before performing a method provided herein. In some embodiments, the evaluation is performed after performing one or more steps of a method provided herein. In some embodiments, the evaluation is performed before administration of one or more steps of a provided combination therapy, e.g., to screen and identify patients suitable and / or susceptible to receiving the combination therapy. In some embodiments, evaluation is performed during, over the course of, or after administration of one or more steps of a provided combination therapy, e.g., to assess intermediate or final treatment outcomes, e.g., to determine activity of the treatment, and / or to decide whether to continue or repeat treatment, and / or to decide whether to administer remaining steps of the combination therapy.

[0380] In some embodiments, the screening step and / or evaluation of treatment outcome includes evaluating the levels and changes in levels, e.g., alterations, of factors, e.g., enzymes, and / or metabolites, in the kynurenine pathway. In some embodiments, the screening step and / or evaluation of treatment outcome includes evaluating immune function, e.g., the immune function of T cells administered for cell therapy and / or endogenous T cells in the body. In some embodiments, the screening step and / or evaluation of treatment outcome includes evaluating the survival and / or function of T cells administered for cell therapy. In some embodiments, the screening step and / or evaluation of treatment outcome includes evaluating cytokine or growth factor levels. In some embodiments, the screening step and / or evaluation of treatment outcome includes evaluating disease burden and / or improvement, e.g., evaluating tumor burden and / or clinical outcome. In some embodiments, any of the screening step and / or evaluation of treatment outcome can include any of the evaluation methods and / or assays described herein and / or known in the art, and can be performed one or more times, e.g., before, during, over the course of, or after administration of one or more steps of the combination therapy.

[0381] In some embodiments, certain parameters can be evaluated to select patients for combination therapy, determine the sequence, dosage, or regimen of combination therapy, assess treatment outcome, and / or determine the activity of combination therapy.In some embodiments, such parameters include, for example, the level of metabolites in the kynurenine pathway, such as TRP, KYN, or KYN derivatives; the expression level of factors or effectors involved in the kynurenine pathway, such as enzymes, for example, IDO1; T cell proliferation; T cell activity; cell phenotype evaluation, such as T cell surface marker expression, and / or the activity of treatment.Evaluation can be performed in vivo, ex vivo, or in vitro, and certain parameters can be determined from biological samples, such as serum samples, plasma samples, tumor biopsies, or by in vivo imaging.Biological samples include, but are not limited to, body fluids such as blood, plasma, serum, cerebrospinal fluid, synovial fluid, urine, and sweat; tissue and organ samples; or tumor samples, including isolated cells and / or processed samples derived from tumors, such as tissue sections from tumor biopsies.

[0382] An exemplary set of parameters associated with treatment outcome that can be assessed in some embodiments of the methods provided herein include plasma kynurenine / tryptophan ratio, peripheral blood immune cell population profile, tumor burden, and plasma tumor markers and immunomodulatory and IDO1. + The expression levels of markers of expression are included.

[0383] A. Screening, Identifying, and / or Selecting Subjects and Determining Dosage and Schedule In some embodiments, subjects can be screened prior to administration of one or more steps of the combination therapy. For example, subjects can be screened for kynurenine pathway factors, e.g., enzyme and / or metabolite levels, availability, concentration, action, activity, metabolism, synthesis, and / or degradation, and / or disease characteristics and / or disease burden, e.g., tumor burden, and / or characteristics or history of prior treatments, and / or response to prior treatments, before or after administration of one or more steps of the combination therapy to determine suitability, responsiveness, and / or sensitivity to the administration of one or more steps of the combination therapy.

[0384] In some embodiments, subjects can be screened for the level, availability, concentration, action, activity, metabolism, synthesis, and / or degradation of kynurenine pathway factors or effectors, e.g., enzymes, and / or metabolites, from a biological sample from the subject, e.g., tumor, blood, and / or serum sample, e.g., the level of a metabolite in the kynurenine pathway, e.g., TRP, KYN, or a derivative of KYN, the expression level of a factor or effector involved in the kynurenine pathway, e.g., an enzyme, e.g., IDO1, and / or an amino acid transporter, e.g., LAT1, LAT2, CD98hc, and / or PAT4. In some embodiments, any of the parameters associated with therapy, screening, identification, or treatment outcome described herein, such as those described in Section III.C, below, can be used as relevant parameters for determining therapeutic suitability, responsiveness, and / or sensitivity, and / or for screening, identifying, and / or selecting subjects for treatment, e.g., using one or more steps of some of the embodiments provided herein, e.g., combination therapy.

[0385] In some embodiments, subjects can be screened after administering one of the steps of combination therapy to determine and identify subjects who will receive the remaining steps of combination therapy.For example, in some embodiments, tryptophan metabolism and / or kynurenine pathway modulator is administered before administering immunotherapy, such as T cell therapy (for example, CAR-expressing T cells) or T cell-binding therapy, and the level of factors of kynurenine pathway, such as enzymes and / or metabolites, is evaluated before administering immunotherapy.In some embodiments, immunotherapy, such as T cell therapy (for example, CAR-expressing T cells) or T cell-binding therapy, is administered before administering tryptophan metabolism and / or kynurenine pathway modulator, and the proliferation and / or activity of administered T cells and / or the level of factors of kynurenine pathway, such as enzymes and / or metabolites, are evaluated before administering tryptophan metabolism and / or kynurenine pathway modulator.In some embodiments, subjects can be screened for specific characteristics of disease, such as tumor or tumor microenvironment (TME) and / or disease burden, such as tumor burden, before administering combination therapy. For example, in some embodiments, subjects who have received T cell therapy, e.g., CAR T cell therapy, are assessed for increased IDO1 expression and / or circulating metabolite levels, such as circulating tryptophan or kynurenine levels, in tumor samples to select and / or identify subjects for administration of a tryptophan metabolism and / or kynurenine pathway modulator. Any of the assessment methods and / or assays described herein and / or known in the art can be performed as a step to screen subjects for combination therapy.

[0386] In some embodiments, provided is a method for selecting a subject with disease or condition for treatment.In some embodiments, provided is a method for evaluating a parameter, for example, any parameter described herein, to screen and / or identify a subject for treatment.In some embodiments, the treatment is administering tryptophan metabolism and / or kynurenine pathway regulator, particularly in combination with T cell therapy.

[0387] In some embodiments, methods for screening, identification, and / or selection include methods for evaluating parameters such as the level and / or amount of factors, molecules, and / or agents that are indicative of or associated with the induction or production of factors, molecules, and / or agents involved in or indicative of adaptive immune resistance, activation of the tryptophan metabolism / kynurenine pathway, IDO1 expression, and / or tryptophan depletion. In some embodiments, such evaluation includes determining the level and / or amount of a parameter such as a factor, molecule, and / or agent, and / or determining a change in the parameter. In some embodiments, the level and / or amount of such a parameter and / or a change in the level and / or amount of a parameter directly correlates or inversely correlates with activation of the tryptophan metabolism / kynurenine pathway, IDO1 expression, and / or tryptophan depletion in a subject.

[0388] In some embodiments, the method includes identifying and / or selecting a subject with a disease or condition for treatment based on the results of the screening and / or assessment. In some embodiments, subjects are selected or identified whose parameter levels and / or amounts and / or changes in parameter levels and / or amounts indicate, for example, directly or inversely correlate with, tryptophan metabolism / kynurenine pathway activation, IDO1 expression, and / or tryptophan depletion. In some embodiments, such subjects are identified and / or selected for administration of a tryptophan metabolism and / or kynurenine pathway modulator and / or T cell therapy.

[0389] In some embodiments, parameters such as levels and / or amounts of factors, molecules and / or agents indicative of or associated with adaptive immune resistance, activation of the tryptophan metabolism / kynurenine pathway, IDO1 expression and / or tryptophan depletion include levels of tryptophan or tryptophan metabolites, levels of expression or activity of IDO1, IDO2 or TDO, or levels of interferon-gamma (IFNγ).

[0390] In some embodiments, a high level of a parameter, such as a level above a threshold level, or an increase in a parameter compared to a previous time point, is associated with or indicates adaptive immune resistance, activation of the tryptophan metabolism / kynurenine pathway, IDO1 expression, and / or tryptophan depletion. In some embodiments, a low level of a parameter, such as a level below a threshold level, or a decrease in a parameter compared to a previous time point, is associated with or indicates adaptive immune resistance, activation of the tryptophan metabolism / kynurenine pathway, IDO1 expression, and / or tryptophan depletion. In some embodiments, if the results of the assessment are associated with or indicate adaptive immune resistance, activation of the tryptophan metabolism / kynurenine pathway, IDO1 expression, and / or tryptophan depletion, the subject is selected for administration of a tryptophan metabolism and / or kynurenine pathway modulator.

[0391] In some embodiments, the parameters can be asse...

Claims

1. 1. A medicament for the treatment of cancer comprising an IDO inhibitor in combination with a composition for T cell therapy comprising a T cell, wherein the IDO inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat), and further wherein (a) the composition is administered to a subject having cancer, wherein the cancer comprises a tumor that comprises cells that are negative for IDO1 prior to the initiation of T cell therapy; and (b) administering to the subject a therapeutically effective amount of an IDO inhibitor within 14 days after initiating administration of the composition; Medicine.

2. A medicament for the treatment of cancer comprising an IDO inhibitor, wherein the IDO inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat), further wherein the medicament is for use in combination with a composition for T cell therapy comprising T cells for treating cancer in a subject, wherein a therapeutically effective amount of the IDO inhibitor is administered to the subject within 14 days after initiation of administration of the composition, further wherein the cancer comprises a tumor comprising cells negative for IDO1 prior to initiation of T cell therapy; Medicine.

3. A medicament for the treatment of cancer comprising an IDO inhibitor, wherein the IDO inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat), further wherein a therapeutically effective amount of the IDO inhibitor is administered to a subject having cancer within 14 days of administration of a composition for T cell therapy comprising T cells, the subject having previously been administered the composition, and further wherein the cancer comprises a tumor comprising cells that are negative for IDO1 prior to the initiation of T cell therapy.

4. The method of any one of claims 1 to 3, wherein the IDO inhibitor is administered within 7 days after administration of the composition.

5. IDO inhibitors prevent or reduce the formation, concentration, availability, metabolism, and / or action of kynurenine pathway enzymes and / or metabolites, and / or IDO inhibitors prevent or reduce the catabolism of tryptophan, prevent or reduce the synthesis or accumulation of tryptophan metabolites, or reduce the ratio of kynurenine to tryptophan; The pharmaceutical composition according to any one of claims 1 to 4.

6. IDO inhibitors at the following times: a time point at which there is a decrease in tryptophan levels, an increase in kynurenine levels, and / or an increase in expression or activity of IDO1, IDO2, or TDO in a biological sample from the subject compared to before administration of the composition began or compared to an earlier time point after administration of the composition began; a time point before the number of detectable cells of the composition in blood from the subject is decreased compared to that in the subject at an earlier time point after initiation of administration of the composition; A time point before T cells exhibit prolonged tryptophan starvation, exhaustion, reduced effector function, expression of inhibitory receptors, and / or loss of proliferative capacity; a time point before the peak or maximum level of cells of the composition is detectable in the blood of the subject; and / or a time point at which the level of interferon-gamma (IFNγ) is increased in a biological sample from the subject compared to before administration of the composition began or compared to an earlier time point after administration of the composition began. The pharmaceutical agent according to any one of claims 1 to 5, which is administered

7. The pharmaceutical composition of any one of claims 1 to 6, wherein the composition comprises tumor infiltrating lymphocytes (TILs) or genetically engineered T cells expressing a recombinant receptor that specifically binds to an antigen.

8. The pharmaceutical of claim 7, wherein the recombinant receptor comprises a chimeric antigen receptor (CAR) or a transgenic T cell receptor (TCR).

9. 9. The method of claim 7 or 8, wherein the recombinant receptor binds to, recognizes or targets an antigen associated with cancer.

10. The antigen is selected from the group consisting of ROR1, Her2, L1-CAM, CD19, CD20, CD22, mesothelin, CEA, hepatitis B surface antigen, antifolate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, EGP-2, EGP-4, EPHa2, ErbB2, ErbB3, ErbB4, FBP, fetal acetylcholine e receptor, GD2, GD3, HMW-MAA, IL-22R-α, IL-13R-α 2, kdr, kappa light chain, Lewis Y, L1 cell adhesion molecule, MAGE-A1, mesothelin, MUC1, MUC16, PSCA, NKG2D ligand, NY-ESO-1, MART-1, gp100, fetal tumor antigen, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), prostate specific antigen, PSMA, estrogen receptor, progesterone receptor, ephrin B2, CD123, CS-1, c-Met, GD-2, MAGE A3, CE7, Wilms' tumor 1 (WT-1), and cyclin A1 (CCNA1).

11. The pharmaceutical composition according to any one of claims 1 to 10, wherein the cancer is a myeloma, lymphoma, leukemia, or non-blood cancer, or a solid tumor.

12. The pharmaceutical agent according to any one of claims 1 to 11, wherein the cancer is non-Hodgkin's lymphoma (NHL), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), or myeloma.

13. The method of any one of claims 1 to 12, wherein the IDO inhibitor is administered orally.

14. IDO inhibitors are inclusive of the following values: 2.5mg-5000mg, 2.5mg-2000mg, 2.5mg-1000mg, 2.5mg-500mg, 2.5mg-200mg, 2.5mg-100mg, 2.5mg-50mg, 2.5mg-25mg, 2.5mg-10mg, 25mg-5000mg, 25mg-2000mg, 25mg-1000mg, 25mg-500mg, 25mg-200mg, 25mg-100mg, 25mg-50mg, 50mg-5000mg, 50mg-200 0mg, 50mg-1000mg, 50mg-500mg, 50mg-200mg, 50mg-100mg, 100mg-5000mg, 100mg-2000mg, 100mg-1000mg, 100mg-500mg, 100mg-200mg, 200mg-5000mg, 200mg-2000mg, 200mg-1000mg, 200mg-500mg, 500mg-5000mg, 500mg-2000mg, 500mg-1000mg or 1000mg-2000mg, the IDO inhibitor is administered six times per day, five times per day, four times per day, three times per day, twice per day, or once per day; and / or the IDO inhibitor is administered at a total daily dose of at least 5 mg / day, 10 mg / day, 25 mg / day, 50 mg / day, 100 mg / day, 200 mg / day, 400 mg / day, 500 mg / day, 600 mg / day, 800 mg / day, 1000 mg / day, 1200 mg / day, 1600 mg / day, 2000 mg / day, 5000 mg / day, or 10000 mg / day; The pharmaceutical composition according to any one of claims 1 to 13.

15. Administration of an IDO inhibitor continues until a time point following initiation of administration of the composition: until there is an increase in tryptophan levels, a decrease in kynurenine levels, and / or a decrease in expression or activity of IDO1, IDO2, or TDO in a biological sample from the subject compared to immediately before administration of the IDO inhibitor or compared to immediately before administration of the composition began; until the number of cells of the composition detectable in blood from the subject is increased compared to that in the subject at an earlier time point immediately prior to administration of the IDO inhibitor or compared to an earlier time point after administration of the composition; until the number of cells of the composition detectable in the blood is within 2.0 times (more or less) the peak or maximum number of cells of the composition detectable in the subject's blood after initiation of administration of the composition; until the number of detectable cells of the composition in blood from the subject exceeds 10%, 15%, 20%, 30%, 40%, 50%, or 60% of the total peripheral blood mononuclear cells (PBMCs) in the subject's blood; The concentration or number of engineered cells in the subject's blood is (i) at least 10 or at least 10 engineered cells per microliter; (ii) at least 20%, 30%, 40%, or 50% of the total number of peripheral blood mononuclear cells (PBMCs); or (iii) at least 1 × 10 5 engineered cells, or (iv) until the subject's blood contains at least 5,000 copies of recombinant receptor-encoding DNA per microgram of DNA; (a) until CAR-expressing cells are detectable in the subject's blood or serum 90 days after the start of administration; and / or (a) 90 days after the start of administration, the subject's blood contains at least 20% CAR-expressing cells, at least 10 CAR-expressing cells per microliter, or at least 1 x 10 4 until it contains CAR-expressing cells. To be continued, The pharmaceutical composition according to any one of claims 1 to 14.

16. The pharmaceutical composition of any one of claims 1 to 15, wherein the composition comprises CD4+ T cells and / or CD8+ T cells.

17. (a) the composition has a concentration of 0.5×10 per kg of the subject's body weight, inclusive; 6 cells / kg ~ 5×10 6 cells / kg, 0.5×10 6 cells / kg ~ 3×10 6 cells / kg, 0.5×10 6 cells / kg ~ 2×10 6 cells / kg, 0.5×10 6 cells / kg ~ 1×10 6 Cells / kg, 1.0 x 10 per kg of subject body weight 6 cells / kg ~ 5×10 6 cells / kg, 1.0×10 6 cells / kg ~ 3×10 6 cells / kg, 1.0×10 6 cells / kg ~ 2×10 6 Cells / kg, 2.0 x 10 per kg of subject body weight 6 cells / kg ~ 5×10 6 cells / kg, 2.0×10 6 cells / kg ~ 3×10 6 cells / kg or 3.0 x 10 cells / kg of subject body weight 6 cells / kg ~ 5×10 6 containing a dose containing many cells / kg, or (b) the composition is 1×10 6 ~3×10 8 Or 1×10 7 ~2×10 8 of total recombinant receptor-expressing cells, optionally 1 x 10 7 , 5×10 7 , 1×10 8 , or 1.5 × 10 8 or a range between any two of the preceding values. The pharmaceutical composition according to any one of claims 1 to 16.

18. The pharmaceutical agent according to any one of claims 1 to 17, which is used in combination with lymphodepleting chemotherapy prior to administration of the composition.

19. The use of a composition for T cell therapy comprising T cells and an IDO inhibitor in the manufacture of a medicament for combined use for the treatment of cancer, wherein the IDO inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat), and further wherein the treatment comprises (a) administering the composition to a subject having cancer, wherein the cancer comprises a tumor that comprises cells that are negative for IDO1 prior to the initiation of T cell therapy; and (b) administering to the subject a therapeutically effective amount of an IDO inhibitor within 14 days after initiating administration of the composition. Including, use.

20. Use of an IDO inhibitor in the manufacture of a medicament for the treatment of cancer, wherein the IDO inhibitor is 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (INCB024360, epacadostat), further comprising: the treatment comprises administering a therapeutically effective amount of an IDO inhibitor to a subject having cancer within 14 days of administration of a composition for T cell therapy comprising T cells, the subject having previously been administered the composition, and the cancer comprises a tumor comprising cells negative for IDO1 prior to the initiation of T cell therapy; use.

21. 21. The use of claim 19 or 20, wherein the IDO inhibitor is administered within 7 days after administration of the composition.

22. IDO inhibitors prevent or reduce the formation, concentration, availability, metabolism, and / or action of kynurenine pathway enzymes and / or metabolites, and / or IDO inhibitors prevent or reduce the catabolism of tryptophan, prevent or reduce the synthesis or accumulation of tryptophan metabolites, or reduce the ratio of kynurenine to tryptophan; 22. Use according to any one of claims 19 to 21.

23. IDO inhibitors at the following times: a time point at which there is a decrease in tryptophan levels, an increase in kynurenine levels, and / or an increase in expression or activity of IDO1, IDO2, or TDO in a biological sample from the subject compared to before administration of the composition began or compared to an earlier time point after administration of the composition began; a time point before the number of detectable cells of the composition in blood from the subject is decreased compared to that in the subject at an earlier time point after initiation of administration of the composition; A time point before T cells exhibit prolonged tryptophan starvation, exhaustion, reduced effector function, expression of inhibitory receptors, and / or loss of proliferative capacity; a time point before the peak or maximum level of cells of the composition is detectable in the blood of the subject; and / or a time point at which the level of interferon-gamma (IFNγ) is increased in a biological sample from the subject compared to before administration of the composition began or compared to an earlier time point after administration of the composition began. The use according to any one of claims 19 to 22, wherein the administration is

24. The use according to any one of claims 19 to 23, wherein said composition comprises tumor infiltrating lymphocytes (TIL) or genetically engineered cells expressing a recombinant receptor that specifically binds to an antigen.

25. 25. The use according to any one of claims 19 to 24, wherein the cancer is a myeloma, lymphoma, leukemia, or non-hematological cancer, or is a solid tumor.

26. 26. The use of any one of claims 19 to 25, wherein the cancer is non-Hodgkin's lymphoma (NHL), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), or myeloma.

27. The use according to any one of claims 19 to 26, wherein the IDO inhibitor is administered orally.

28. IDO inhibitors are inclusive of the following values: 2.5mg-5000mg, 2.5mg-2000mg, 2.5mg-1000mg, 2.5mg-500mg, 2.5mg-200mg, 2.5mg-100mg, 2.5mg-50mg, 2.5mg-25mg, 2.5mg-10mg, 25mg-5000mg, 25mg-2000mg, 25mg-1000mg, 25mg-500mg, 25mg-200mg, 25mg-100mg, 25mg-50mg, 50mg-5000mg, 50mg-200 0mg, 50mg-1000mg, 50mg-500mg, 50mg-200mg, 50mg-100mg, 100mg-5000mg, 100mg-2000mg, 100mg-1000mg, 100mg-500mg, 100mg-200mg, 200mg-5000mg, 200mg-2000mg, 200mg-1000mg, 200mg-500mg, 500mg-5000mg, 500mg-2000mg, 500mg-1000mg or 1000mg-2000mg, the IDO inhibitor is administered six times per day, five times per day, four times per day, three times per day, twice per day, or once per day; and / or the IDO inhibitor is administered at a total daily dose of at least 5 mg / day, 10 mg / day, 25 mg / day, 50 mg / day, 100 mg / day, 200 mg / day, 400 mg / day, 500 mg / day, 600 mg / day, 800 mg / day, 1000 mg / day, 1200 mg / day, 1600 mg / day, 2000 mg / day, 5000 mg / day, or 10000 mg / day; 28. Use according to any one of claims 19 to 27.

29. Administration of an IDO inhibitor continues until a time point following initiation of administration of the composition: until there is an increase in tryptophan levels, a decrease in kynurenine levels, and / or a decrease in expression or activity of IDO1, IDO2, or TDO in a biological sample from the subject compared to immediately before administration of the IDO inhibitor or compared to immediately before administration of the composition began; until the number of cells of the composition detectable in blood from the subject is increased compared to that in the subject at an earlier time point immediately prior to administration of the IDO inhibitor or compared to an earlier time point after administration of the composition; until the number of cells of the composition detectable in the blood is within 2.0 times (more or less) the peak or maximum number of cells of the composition detectable in the subject's blood after initiation of administration of the composition; until the number of detectable cells of the composition in blood from the subject exceeds 10%, 15%, 20%, 30%, 40%, 50%, or 60% of the total peripheral blood mononuclear cells (PBMCs) in the subject's blood; The concentration or number of engineered cells in the subject's blood is (i) at least 10 engineered cells per microliter; (ii) at least 20%, 30%, 40%, or 50% of the total number of peripheral blood mononuclear cells (PBMCs); or (iii) at least 1 × 10 5 engineered cells, or (iv) until the subject's blood contains at least 5,000 copies of recombinant receptor-encoding DNA per microgram of DNA; (a) until CAR-expressing cells are detectable in the subject's blood or serum 90 days after the start of administration; and / or (a) 90 days after the start of administration, the subject's blood contains at least 20% CAR-expressing cells, at least 10 CAR-expressing cells per microliter, or at least 1 x 10 4 until it contains CAR-expressing cells. To be continued, 29. The use according to any one of claims 19 to 28.

30. The use according to any one of claims 19 to 29, wherein the composition comprises CD4+ T cells and / or CD8+ T cells.

31. (a) the composition has a concentration of 0.5×10 per kg of the subject's body weight, inclusive; 6 cells / kg ~ 5×10 6 cells / kg, 0.5×10 6 cells / kg ~ 3×10 6 cells / kg, 0.5×10 6 cells / kg ~ 2×10 6 cells / kg, 0.5×10 6 cells / kg ~ 1×10 6 Cells / kg, 1.0 x 10 per kg of subject body weight 6 cells / kg ~ 5×10 6 cells / kg, 1.0×10 6 cells / kg ~ 3×10 6 cells / kg, 1.0×10 6 cells / kg ~ 2×10 6 Cells / kg, 2.0 x 10 per kg of subject body weight 6 cells / kg ~ 5×10 6 cells / kg, 2.0×10 6 cells / kg ~ 3×10 6 cells / kg or 3.0 x 10 cells / kg of subject body weight 6 cells / kg ~ 5×10 6 containing a dose containing many cells / kg, or (b) the composition is 1×10 6 ~3×10 8 Or 1×10 7 ~2×10 8 of total recombinant receptor-expressing cells, optionally 1 x 10 7 , 5×10 7 , 1×10 8 , or 1.5 × 10 8 or a range between any two of the preceding values. Use according to any one of claims 19 to 30.

32. The use according to any one of claims 19 to 31, wherein the treatment further comprises the administration of lymphodepleting chemotherapy prior to administration of said composition.

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