BCL-2 inhibition to amplify chimeric antigen receptor therapy

Abstract

Provided herein are methods of preparing an immunotherapeutic with enhanced efficacy by treating lymphocytes with a B-cell lymphoma 2 (BCL-2) inhibitor, immunotherapeutic compositions produced by the methods herein, and methods of treating cancer therewith.

Description

[0001]Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 BCL-2 INHIBITION TO AMPLIFY CHIMERIC ANTIGEN RECEPTOR THERAPY CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 596,189, filed on November 3, 2023, which is incorporated by reference herein. FIELD Provided herein are methods of preparing an immunotherapeutic with enhanced efficacy by treating lymphocytes with a B-cell lymphoma 2 (BCL-2) inhibitor, immunotherapeutic compositions produced by the methods herein, and methods of treating cancer therewith. BACKGROUND Immune-based therapies including immune checkout blockade and adoptive T-cell therapies (e.g., using T cells engineered to express CD19-specific chimeric antigen receptor) have provided recent breakthroughs in treating certain cancers (e.g., hematologic malignancies). Despite great advances in cancer immunotherapies, durable success of these treatments has been limited to a fraction of patients and cancer types. Strategies to enhance the efficacy of immunotherapies are urgently needed to deliver long-term therapeutic outcomes to a greater number of patients with a broader array of tumor types. SUMMARY Provided herein are methods of preparing an immunotherapeutic with enhanced efficacy by treating lymphocytes with a B-cell lymphoma 2 (BCL-2) inhibitor, immunotherapeutic compositions produced by the methods herein, and methods of treating cancer therewith. In some embodiments, provided herein are immunotherapeutic compositions comprising a lymphocyte (i) engineered to express a chimeric antigen receptor (CAR), and (ii) treated with a B-cell lymphoma 2 (BCL-2) inhibitor. In some embodiments, provided herein are methods of enhancing a chimeric antigen receptor (CAR) lymphocyte comprising treating the CAR lymphocyte with a B-cell lymphoma 2 (BCL-2) inhibitor. In some embodiments, an immunotherapeutic composition is provided Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 comprising a chimeric antigen receptor (CAR) lymphocyte enhanced by the methods described herein. In some embodiments, provided herein are methods of preparing an immunotherapeutic composition comprising: (a) obtaining lymphocytes from a subject; (b) activating the lymphocytes; (c) inducing the lymphocytes to express a chimeric antigen receptor (CAR) to produce CAR lymphocytes; and (d) treating the CAR lymphocytes with a B-cell lymphoma 2 (BCL-2) inhibitor. In some embodiments, immunotherapeutic compositions prepared by the methods described herein are provided. In some embodiments, provided herein are immunotherapeutic compositions wherein the CAR comprises an extracellular antigen recognition domain capable of binding to a target selected from the group consisting of TSHR, CD19, CD123, CD22, BAFF-R, CD30, CD171, CS-1, CLL-1, CD33, EGFRvIII, GD2, GD3, BCMA, GPRC5D, Tn Ag, PSMA, ROR1, FLT3, FAP, TAG72, CD38, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, mesothelin, IL-11Ra, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-0, SSEA-4, CD20, AFP, Folate receptor α, ERBB2 (Her2 / neu), MUC1, EGFR, CS1, CD138, NCAM, Claudin18.2, Prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gp100, bcr-ab1, tyrosinase, EphA2, Fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, Folate receptor R, TEM1 / CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD 179a, ALK, polysialic acid, PLAC1, GloboH, NY- BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-1a, MAGE-A1, legumain, HPV E6 / E7, MAGE A1, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, prostate specific protein, survivin and telomerase, PCTA-1 / Galectin 8, MelanA / MART1, Ras mutant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2ETS fusion gene), NA17, PAX3, androgen receptor, Cyclin B1, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES 1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal tract carboxylesterase, mut hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, PD1, PDL1, PDL2, TGFβ, APRIL, and NKG2D. In some embodiments, the extracellular antigen recognition domain capable of binding to CD19. In some embodiments, the extracellular antigen recognition domain is a short-chain variable fragment (scFv). In some embodiments, the CAR further comprises a transmembrane domain and intracellular signaling domain. In some embodiments, Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 the CAR further comprises a costimulatory domain. In some embodiments, in methods or compositions herein, the BCL-2 inhibitor is a BCL- 2 homology 3 (BH3) mimetic. In some embodiments, the BH3 mimetic is selected from venetoclax, ABT-199 (4-[4-[[2-(4-Chlorophenyl)-4,4-dimethylcyclohex-1-en-1- yl]methyl]piperazin-1-yl]-N-[[3-nitro-4-[[(tetrahydro-2H-pyran-4- yl)methyl]amino]phenyl]sulfonyl]-2-[(1H-pyrrolo[2,3-b]pyridin-5-yl)oxy]benzamide), ABT-737 (4-[4-[[2-(4-chlorophenyl)phenyl]methyl]piperazin-1-yl]-N-[4-[[(2R)-4-(dimethylamino)-1- phenylsulfanylbutan-2-yl] amino]-3-nitrophenyl]sulfonylbenzamide) (navitoclax), ABT-263 ((R)-4-(4-((4′-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1′-biphenyl]-2-yl)methyl)piperazin-1- yl)-N-((4-((4-morpholino-1-(phenylthio)butan-2-yl)amino)- 3((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide), GX15-070 (obatoclax mesylate, (2Z)- 2-[(5Z)-5-[(3,5-dimethyl-1H-pyrrol-2-yl)methylidene]-4-methoxypyrrol-2-ylidene]indole; methanesulfonic acid))), 2-methoxy-antimycin A3, YC137 (4-(4,9-dioxo-4,9- dihydronaphtho[2,3-d]thiazol-2-ylamino)-phenyl ester), pogosin, ethyl 2-amino-6-bromo-4-(1- cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate, Nilotinib-d3, TW-37 (N-[4-[[2-(1,1- Dimethylethyl)phenyl]sulfonyl]phenyl]-2,3,4-trihydroxy-5-[[2-(1- methylethyl)phenyl]methyl]benzamide), Apogossypolone (ApoG2), HA14-1, AT101 (gossypol), sabutoclax, gambogic acid, and G3139 (Oblimersen). In some embodiments, In some embodiments, in methods or compositions herein, the BCL-2 inhibitor inhibits expression of BCL-2. In some embodiments, the BCL-2 inhibitor is an shRNA, a miRNA, a morpholino, a ribozyme, an antisense nucleic acid molecule, or a CRISPR-Cas9-based construct. In some embodiments, in methods or compositions herein, the lymphocyte(s) are T cells. In some embodiments, in methods or compositions herein, the lymphocyte(s) are NK cells. In some embodiments, provided herein are methods of treating a subject suffering from cancer comprising administering to the subject the immunotherapeutic composition described herein. In some embodiments, the subject suffers from a blood or bone marrow cancer. In some embodiments, the subject suffers from a leukemia, lymphoma or myeloma. In some embodiments, the subject suffers from B-cell acute lymphoblastic leukemia (B-ALL), chronic lymphocytic leukemia (CLL), or B-cell non-Hodgkin lymphoma (B-NHL), In some embodiments, the subject suffers from a solid tumor cancer. In some embodiments, methods further comprise co-administering one or more additional cancer treatments. In some Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 embodiments, the one or more additional cancer treatments are selected from chemotherapies, immunotherapeutics, radiation treatment, and surgery. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1A-C. Venetoclax does not affect T cell expansion and upregulates anti-apoptotic protein expression in T cells. T cells were isolated from PBMCs of a healthy donor on day 0 and activated overnight with human anti-CD3 / CD28 beads, followed by transduction with lentivirus designed to express CD19CAR on day 1. On day 2, cells were either treated with vehicle (DMSO) or with the indicated doses of venetoclax daily until day 13. On day 13, cells were harvested, counted and intracellularly stained for BCL-2 family protein expression. As shown in (A) Venetoclax pre-treated hCD19CART at various doses show no significant change in T cell expansion compared to vehicle (DMSO)-pretreated hCD19CART. (B) hCD19CART show increased anti-apoptotic protein expression upon expansion in presence of venetoclax. (C) Venetoclax-pretreated hCD19CARTs show slight, but inconsistent, change in CART memory phenotype. Fig. 2A-B. Venetoclax enhances hCD19CART cytotoxicity in vitro. (A) Flow-based killing assay. Human diffuse large B cell lymphoma (DLBCL) OCI-Ly8.Chili tumor cells were co-cultured with vehicle or venetoclax pre-treated hCD19CAR T cells in different effector: tumor (E:T) ratios. Killing was assessed by flow cytometry at different time points. (B) Bioluminescent imaging (BLI)-based cytotoxic assay. BLI image (on the left) shows 96 well plate seeded with untreated (NoTx), vehicle (DMSO) or venetoclax-pretreated hCD19CART harvested on day 13 after expansion and co-cultured with OCI-Ly8.Luci tumor cells (expressing luciferase enzyme) in E:T = 1:5. After 48 hour - incubation, luciferin was added to the culture and the presence of tumor cells was measured by BLI. The bar graph shows that venetoclax enhances the killing potency of pre-treated hCD19CART in a dose dependent manner. (Note: Venetoclax was not present in any assay). Fig. 3A-B. Venetoclax pre-treated hCD19CAR T cells exhibit enhanced cytotoxicity through generation of polyfunctional T cells capable of generating multiple cytokines and death inducer molecules. hCD19CART prepared in the absence or presence of increasing doses of venetoclax were co-cultured with OCI-Ly8.Luci tumor cells in E:T = 1:5. Upon incubation for 48 hours, cells were harvested for FACS staining as in (A) and the supernatant was collected for Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 detection of cytokines and effector molecules as in (B). (A) Venetoclax pre-treated hCD19CAR T cells show upregulated expression of TNF-related apoptosis-inducing ligand (TRAIL) and FAS in addition to enhanced IFNg production compared to vehicle (DMSO)-pretreated hCD19CART. (B) Venetoclax pre-treated hCD19CAR T cells demonstrate enhanced cytokine and effector molecule production compared to vehicle (DMSO)-pretreated hCD19CAR T cells when co-cultured with OCI-Ly8 tumor cells. Fig. 4. Venetoclax effect on hCD19CART is independent on the costimulatory molecule used in the CART vector. hCD19CART cells were manufactured with constructs that include CD28 or 4-1BB costimulatory regions. hCD19CART cells were expanded and tested for their ability to kill human DLBCL as described in Fig. 2. hCD19CART cells were incubated with human DLBCL at effector to target ratios of 1:3 and 1:5 and tumor cell death was measured at 24 hrs and 48 hours after co-incubation. Fig. 5. In vitro stress (antigen-challenge) assay. hCD19CART cells expanded in the presence of vehicle (DMSO) or 800nM venetoclax were cultured with CD19+DLBCL. CAR T cells were harvested and re-plated in new culture with fresh CD19+DLBCL tumor cells every two to three days. Flow cytometric analysis of hCD19CART cells was performed following each stimulation for expression of markers of T cell exhaustion including LAG3, PD-1, and TIM3. Both CD4+and CD8+hCD19CART cells expanded in venetoclax following 1stand 3rdantigen stimulations had decreased expression of exhaustion markers. Fig. 6. Venetoclax-induced transcriptional alterations in hCD19CART. After expansion of hCD19CART cells in presence or absence of venetoclax, cells were subjected to single cell RNA sequencing (scRNAseq). (A) Venn diagram of the number of significantly upregulated (red) and downregulated (blue) genes in CD8 T cells in response to treatment. BCL-2 transcript / gene expression is markedly increased in venetoclax pre-treated CD19CART (at the bottom). (B) Gene expression heatmap (on the top) of differentially expressed genes (log- fold change > 0.3 and adjusted p value < .05) between all vehicle or venetoclax pre-treated and untreated hCD19CART cells.. Fig. 7. Gene-set enrichment analysis (GSEA) of CD4+and CD8+hCD19CART cells expanded in venetoclax shows expression enrichment of TNFα and IFNγ signaling pathways. Fig. 8. Gene-set enrichment analysis (GSEA) of CD4+and CD8+hCD19CART cells expanded in venetoclax shows expression enrichment of IL-2 / STAT5 and PI3K / AKT signaling Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 pathways. This was supported by increased pAKT and pSTAT5 CD4+and CD8+hCD19CART cells as shown by flow cytometry. Fig. 9. Venetoclax augments the efficacy of hCD19CART in vivo. NSG immunocompromised animals were challenged with subcutaneous injection of OCI-Ly8 DLBCL. When tumors reached 50mm3, mice were treated with vehicle (black line) hCD19CART cells expanded in vehicle alone (blue line) or hCD19CART cells expanded in venetoclax (red line). Tumor volume (right) and survival (left) were measured over time. Fig. 10. NSG mice challenged with disseminated (IV injection) of hDLBCL (OCI-Ly8) followed by injection of hCD19CART were sacrificed and persistence of hCD19CART in the spleens of these animals was evaluated using flow cytometry. While all mice had evidence of disease 28 days following challenge, those animals treated with hCD19CART expanded in venetoclax showed >17x fold expansion compared to hCD19CART expanded in vehicle (DMSO). Fig. 11A-G. Venetoclax shows no effects on T cell expansion, content or memory profile, but upregulates anti-apoptotic protein expression in CAR T cells. (A) The schema depicts the timeline of CART production. T cells were isolated from PBMCs of a healthy donor on day 0 and activated overnight with human anti-CD3 / CD28 beads, followed by transduction with lentivirus designed to express CD19CAR on day 1. On day 2, cells were either treated with vehicle (DMSO) or with the indicated doses of venetoclax daily until day 13. (B) Venetoclax- pretreated hCD19CARTs at various doses show no significant change in T cell expansion compared to vehicle-pretreated hCD19CARTs. On left: proliferation curve indicating CART cell count under each treatment condition at different time points. On right: comparing and summarizing the cell count of CARTs under different conditions on day 10 of expansion. (C, D) hCD19CARTs show increased anti-apoptotic protein expression upon expansion in presence of venetoclax. On day 13, cells were harvested and intracellularly stained for BCL-2 family protein expression. (C) Showing BCL-2, BCLXL and MCL-1 histogram overlay of venetoclax- pretreated hCD19CARTs (red) and vehicle-pretreated hCD19CARTs (black). (D) BCL-2 family protein expression was determined by Mean fluorescence intensity (MFI) of venetoclax- pretreated hCD19CARTs or vehicle-pretreated hCD19CARTs normalized to vehicle-pretreated hCD19CARTs. (E-G) Venetoclax-pretreated hCD19CARTs show no significant changes in hCD19CART composition or memory phenotype. No significant difference was detected Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 between Venetoclax-pretreated hCD19CARTs in (E) CD4:CD8 ratio, (F) transduction efficacy detected by CAR+GFP expression or (G) T cell memory phenotype. TN: naïve (CCR7+CD45RA+), TCM: central memory (CCR7+CD45RA-), TEM: effector memory (CCR7- CD45RA-), and TEMRA: terminally differentiated (CCR7-CD45RA+). Data are summarized from at least three independent experiments from different healthy donors (n=3) with two-three technical replicates in bar graphs and shown as mean ± SD. One-way ANOVA followed by Tukey’s multiple comparison tests was used for (B), unpaired Student’s t-test for (D-F) and Two-way ANOVA followed by SIDAK's multiple comparison tests for (G). *P < 0.05, **P<0.01, ***p<0.001, ****p<0.0001, ns: non-significant. Fig. 12A-G. Venetoclax potentiate hCD19CART cytotoxicity in vitro via increased cytokines and cell death- and cell exhaustion- resistance. (A, B) BLI-based cytotoxic assay using a bioluminescent imaging (BLI) system. BLI image shows 96 well plate seeded with either tumor alone, vehicle (DMSO)- pretreated hCD19CARTs or hCD19CARTs pretreated with escalating doses of venetoclax harvested on day13 and co-cultured with (A) human diffuse large B cell lymphoma (DLBCL) OCI-Ly8.Luci tumor cells (expressing luciferase enzyme) or (B) Leukemia: NALM6 tumors in E:T = 1:5. After 48-hour incubation, luciferin was added to the culture and the presence of tumor cells was measured by BLI. The bar graph shows that venetoclax enhances the killing potency of pretreated hCD19CARTs in a dose dependent manner. (C-G) venetoclax enhances the effector function of CARTs. After 48-hour incubation with OCI-Ly8.Luci tumor cells, (C) supernatant was collected for ELISA and venetoclax- pretreated hCD19CARTs demonstrate enhanced cytokine and effector molecule release compared to vehicle-pretreated hCD19CARTs. (D) Memory profiling shows no significant difference in CD4+CAR T cell phenotype and indicates accumulation of TCM in venetoclax- pretreated CD8+CARTs while more TEMare detected in vehicle-pretreated CD8+CARTs. (E) Venetoclax-pretreated hCD19CARTs demonstrate longer persistence marked by a reduced tendency to undergo apoptosis. (F) Venetoclax-pretreated CD8+CARTs upregulated expression of TNF-related apoptosis-inducing ligand (TRAIL) and increased intracellular IFNγ upon tumor stimulation. (G) Additionally, venetoclax-pretreated hCD19CAR T cells exhibit less exhausted CAR T cell phenotype after two days incubation with tumors. Data represent mean ± SD. Statistics for (A-C) is one-way ANOVA with post hoc Tukey’s tests was performed and two-way ANOVA with SIDAK's multiple comparison tests for (D). *P < 0.05, ** P < 0.01, *** P < Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 0.001, and ****P < 0.0001. (A-C, E-G) Results shown are from single experiment representative of at least three independent experiments with similar results using CARTs of three healthy donors. Fig. 13A-G. Venetoclax improves hCD19CART antitumor activities in a BCL-2 dependent manner. (A-E) Using the a CD19CAR or a newly designed CD19CAR constructs that overexpress the wildtype or mutant BCL-2, T cells were transduced and treated as indicated in schema Fig. 1A. Different constructs pretreated with vehicle are indicated by empty symbol and dashed line, while venetoclax-pretreated hCD19CARTs at dose 800nM have filled symbol and solid connecting lines. (A) Proliferation curves summary indicates CART cell count under each construct condition at different time points. Comparing and summarizing the cell count of CARTs of each CD19CAR construct on day 10 of expansion demonstrate no significant changes in expansion between vehicle vs venetoclax-pretreated CARTs. (B-D) Anti-apoptotic Bcl-2 family protein expression in different BCL-2 overexpressing CARTs as measured by intracellular staining, analyzed by flow cytometry and summarized and normalized into MFI fold change from two healthy donors. CARTs were first gated on the GFP positive fraction, and untraduced T cells (UTDs) were ungated. (B) BCL-2 expression was upregulated in all CARTs with WT or mutant BCL-2 and further upregulated to different extent after treatment with venetoclax. (C, D) CARTs with BCL-2 mutant show limited to no increase in BCLXL and MCL-1 expression upon expansion in presence of venetoclax. (E) CAR transduction, detected by gating GFP+, is homogenous and comparable among T cells transduced with different CAR19 and CAR19_BCL-2 constructs that either pretreated with vehicle or venetoclax. (F) CD19CARTs expressing BCL-2 mutants show attenuation of venetoclax-induced antitumor activity compared to venetoclax-pretreated CD19CARTs expressing wild-type BCL-2. (G) Venetoclax-pretreated CARTs with BCL-2 mutants show gradual loss of venetoclax ability to resist exhaustion. (A-D) Data are from single experiment of CARTs prepared from two healthy donors with three technical replicates and MFI normalized to the corresponding vehicle- pretreated donor’s CART. (E) Data summarized from 3 independent experiments with 3 healthy donor samples and three technical replicates each. (F, G) Data from one experiment that represent similar results of 3 independent experiments. (A-F) Data shown as mean ± SD. Two- Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 way ANOVA with SIDAK's multiple comparison tests was performed. *P < 0.05, **P<0.01, ***p<0.001, ****p<0.0001, ns: non-significant. Fig. 14A-G. Transcriptomic analysis of venetoclax-pretreated CARTs reveal ubiquitous transcriptional reprogramming in several T cell signaling, proliferative, and metabolic pathways. Vehicle or venetoclax-pretreated CARTs were generated as in Fig. 11A, sorted for live cells and analyzed by scRNA-seq on day 13 after expansion without tumor stimulation. The experiment was performed in three biological replicates. (A) Principal component analysis (PCA) of VST transformed counts in DESeq2 for bulk RNA-seq, of CD4+ and CD8+ CART-cells. (B) Uniform manifold approximation and projection (UMAP) embedding of clusters in CD4+ and CD8+ CART cells in scRNAseq. Clustering and UMAP performed on harmony reductions integrated for donor and treatment variables. (C) Proportion of each cluster within its respective treatment group, with dots indicating individual donors. (D) BCL-2 expression level between VX800 and vehicle within each cluster. (E) Heatmap of pseudo-bulk Z-score scaled expression of DEGs with BH-adjusted p-value<0.05 avg_log2FC>0.58. (F) GSEA results against HALLMARK database of pseudo-bulked scRNA-seq. Genes were ranked by the DESeq2 Wald test statistic and ran through cluster Profiler’s GSEA implementation. (G) Cluster-wise GSEA results of selected HALLMARK pathways in scRNA-seq data. Counts for each cluster were pseudo bulked and genes were ranked by DESeq2 Wald test statistic before running cluster-Profiler’s GSEA implementation. (H) Most frequent leading-edge genes across common significantly enriched pathways in bulk RNA-seq and scRNA-seq GSEA against HALLMARK pathways with genes ranked according to DESeq2. (C, D) Data shown as mean ± SD. Two-way ANOVA with SIDAK's multiple comparison tests was performed. ****p<0.0001, ns: non-significant. Fig. 15A-J. Venetoclax improves CAR T cell signaling and metabolic fitness. (A, C, G) Enrichment plots of GSEA of select HALLMARK pathways; genes ranked by DESeq2 Wald test-statistic in pseudobulked scRNA-seq data. (B, D) Venetoclax-pretreated hCD19CARTs demonstrate increased intracellular protein expression of phospho- STAT5 (Tyr694) and phosphor-AKT (Ser473). (E, F) Vehicle or venetoclax-pretreated hCD19CARTs prepared as indicated in Fig.1A were harvested on day 13 and incubated overnight with STAT5 inhibitor (E) or escalating doses of AKT inhibitor (F). Next day, hCD19CARTs were washed from inhibitors, counted and used to set up killing assay. hCD19CARTs treated with or without inhibitors were co-cultured with OCI-Ly8.Luci tumors in effector to tumor (E:T) ratio (1:5) and incubated for 48 Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 hours before BLI imaging. (H-J) Analysis of vehicle or venetoclax-pretreated hCD19CARTs oxidative consumption and glycolysis by Seahorse Mito Stress and Glycolysis Stress assays. Data represent analysis of n = 8 technical replicates from n =2 donors of n=2 independent experiments in mean ± SD. OCR: oxygen consumption rate. ECAR: extracellular acidification rate. Fig. 16A-I. Venetoclax augments the efficacy of hCD19CART prepared from healthy donor or patient samples in vivo. (A-C) Following the experiment’s schema, NSG mice were intravenously injected with NALM6 tumors. After three days, mice were randomly divided into three groups that either received no treatment (No Tx) or treated with vehicle or venetoclax- pretreated hCD19CARTs. Then, mice were subject to bioluminescence imaging at different time points after immediately intraperitonially injected with D-luciferin (150mg / kg) to monitor tumor growth by tracking tumor signal intensity and following mice survival. (D-E) Following the indicated experiment timeline, NSG mice with established subcutaneous (SQ) OCI-Ly8 tumors (25-50mm2) received the indicated treatments. Mice received venetoclax-treated hCD19CART show better tumor control and longer survival. (A-E) The results are pooled for analysis from at least two independent experiments. The number of mice in each group is given. Mouse survival is summarized in the Kaplan-Meier plot. (F-I) PBMCs from patients at the time of commercial CAR T cell apheresis were used to isolate T cells and prepare vehicle or venetoclax-pretreated hCD19CARTs following the method indicated in Fig.1A. Vehicle or venetoclax-pretreated hCD19CARTs prepared from patient apheresis were used to setup killing assay as in (F) or to treat NSG mice bearing subcutaneous OCI-Ly8 tumors (G-I). Mouse survival is summarized in the Kaplan-Meier plot. The number of mice in each group is given. Data shown as mean ± SD. *P < 0.05, **P<0.01, ***p<0.001, ****p<0.0001, ns: non-significant. Fig. 17A-E. Venetoclax directly enhances the effector function of murine CD19CARTs. (A) Schematic depicting experimental setp. (B) Administration of venetoclax either in vivo, ex vivo or both demonstrates enhanced antitumor cytotoxicity compared to vehicle only condition. (C) Short-term treatment of murine T cells with venetoclax for 2 days during CART manufacturing is enough to exhibit superior tumor killing effect over vehicle-treated CARTs. (D / E) CD19CAR expression and the memory profile of the CART product after the short-term treatment with vehicle or venetoclax. Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 Fig. 18A-D. (A / B) Minimum doses of A-1331852, a BCL-XLInhibitor, or ABT-263 (navitoclax) results in low expansion of hCD19CART. (C) A-133 and navitoclax-pretreated hCD19CARTs show increase in the BCL-2 expression but to less extent compared to the same venetoclax dose. (D) hCD19CARTs preconditioning with Navitoclax at low dose 25nM show enhanced antitumor effect. However, pretreatment of hCD19CARTs with other BH3 mimetics such as A-133 is unable to enhance their killing efficacy. Fig. 19A-D. (A) cytotoxicity assay was repeated using CD19 knock out OCI-Ly8 tumor cells (OCI-Ly8.CD19KO.Luci). (B) Venetoclax-pretreated hCD19CARTs fail to kill the CD19- cell line. Venetoclax-pretreated hCD19CARTs induced similar enhanced cytotoxicity in vitro regardless the type of the co-stimulatory molecule in the CD19CAR construct (C), or the cytokine used in the CAR T cell expansion (D). Fig. 20A-D. Gene set enrichment analysis (GSEA) has also demonstrated the upregulation of transcriptomic signature of IFN-α, IFN-γ and TNF-α signaling. Fig. 21A-B. (A) Incubating venetoclax-pretreated CARTs with escalating doses of the antioxidant N-Acetyl-L-cysteine (LNAC) for 6 or 24 hours before co-culturing with tumor shows no effect on the venetoclax-induced killing capacity of CARTs. (B) This result demonstrates that ROS induction is not involved in the observed effect of ventoclax on CARTs in the system. Fig. 22A-H. Venetoclax augments the efficacy of hCD19CARTs prepared from patient apheresis bulb in vivo. (A) Most of CR-samples contain the highest percent of CD4 and consequently the lowest percent of CD8 while most of PR-samples show higher percent of CD8 T cells. (B, F, G) T cell memory profiling reveals that HD-samples tend to possess T cells in naïve (TN), to central memory (TCM) status, PR-samples tend to have cells in a more differentiated less naïve status while CR-samples memory status fall in the middle between HD and PR-samples which also reflected by the expression level of CD62L,CD127 and TCF1 HD > CR > PR. (C) PR-samples contains % of Tregs comparable to HD but higher than the CR- samples. (D, E) Most of exhaustion surface markers like LAG3 and TIM3 and the intracellular marker TOX indicate that PR-samples contain more exhausted cells than HD and CR-samples. (H) CR-samples show the highest expression level of the anti-apoptotic proteins BCL-2, BCL- XL and MCL-1. Fig. 23A-C. Venetoclax augments the efficacy of hCD19CARTs prepared from patient apheresis bulb in vivo. (A) Transduction efficiency was comparable between HD and CR- Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 samples, but PD-samples show lower transduction efficiency. (B) While HD-samples show higher content of CD8+T cells and CR-samples has the lowest content of CD8+reflecting the starting material status, PR-samples show unexpected shrink of CD8+population in favor of more expanded CD4+T cells. (C) Tregs content was comparable between CR- and PR-samples which is slightly higher than HD-samples. DEFINITIONS Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments described herein, some preferred methods, compositions, devices, and materials are described herein. However, before the present materials and methods are described, it is to be understood that this invention is not limited to the particular molecules, compositions, methodologies or protocols herein described, as these may vary in accordance with routine experimentation and optimization. It is also to be understood that the terminology used in the description is for the purpose of describing the particular versions or embodiments only, and is not intended to limit the scope of the embodiments described herein. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. However, in case of conflict, the present specification, including definitions, will control. Accordingly, in the context of the embodiments described herein, the following definitions apply. As used herein and in the appended claims, the singular forms “a”, “an” and “the” include plural reference unless the context clearly dictates otherwise. Thus, for example, reference to “a BCL-2 inhibitor” is a reference to one or more BCL-2 inhibitors and equivalents thereof known to those skilled in the art, and so forth. As used herein, the term “comprise” and linguistic variations thereof denote the presence of recited feature(s), element(s), method step(s), etc. without the exclusion of the presence of additional feature(s), element(s), method step(s), etc. Conversely, the term “consisting of” and linguistic variations thereof, denotes the presence of recited feature(s), element(s), method step(s), etc. and excludes any unrecited feature(s), element(s), method step(s), etc., except for ordinarily-associated impurities. The phrase “consisting essentially of” denotes the recited feature(s), element(s), method step(s), etc. and any additional feature(s), element(s), method Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 step(s), etc. that do not materially affect the basic nature of the composition, system, or method. Many embodiments herein are described using open “comprising” language. Such embodiments encompass multiple closed “consisting of” and / or “consisting essentially of” embodiments, which may alternatively be claimed or described using such language. As used herein, the term “pharmaceutically acceptable carrier” refers to non-toxic solid, semisolid, or liquid filler, diluent, encapsulating material, formulation auxiliary, excipient, or carrier conventional in the art for use with a therapeutic agent for administration to a subject. A pharmaceutically acceptable carrier is non-toxic to recipients at the dosages and concentrations employed and is compatible with other ingredients of the formulation. The pharmaceutically acceptable carrier is appropriate for the formulation employed. For example, if the therapeutic agent is to be administered orally, the carrier may be a gel capsule. A “pharmaceutical composition” typically comprises at least one active agent (e.g., the copolymers described herein) and a pharmaceutically acceptable carrier. As used herein, the term “effective amount” refers to the amount of a composition (e.g., pharmaceutical composition) sufficient to effect beneficial or desired results. An effective amount can be administered in one or more administrations, applications or dosages and is not intended to be limited to a particular formulation or administration route. As used herein, the term “administration” refers to the act of giving a drug, prodrug, or other agent, or therapeutic treatment (e.g., pharmaceutical compositions of the present invention) to a subject or in vivo, in vitro, or ex vivo cells, tissues, and organs. Exemplary routes of administration to the human body can be through the eyes (e.g., intraocularly, intravitreally, periocularly, ophthalmic, etc.), mouth (oral), skin (transdermal), nose (nasal), lungs (inhalant), oral mucosa (buccal), ear, rectal, by injection (e.g., intravenously, subcutaneously, intratumorally, intraperitoneally, etc.) and the like. As used herein, the terms “co-administration” and “co-administer” refer to the administration of at least two agent(s) or therapies to a subject. In some embodiments, the co- administration of two or more agents or therapies is concurrent (e.g., in the same or separate formulations). In other embodiments, a first agent / therapy is administered prior to a second agent / therapy. Those of skill in the art understand that the formulations and / or routes of administration of the various agents or therapies used may vary. The appropriate dosage for co- administration can be readily determined by one skilled in the art. In some embodiments, when Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 agents or therapies are co-administered, the respective agents or therapies are administered at lower dosages than appropriate for their administration alone. Thus, co-administration is especially desirable in embodiments where the co-administration of the agents or therapies lowers the requisite dosage of a potentially harmful (e.g., toxic) agent(s). As used herein, the term “subject” broadly refers to any animal, including but not limited to, human and non-human animals (e.g., dogs, cats, cows, horses, sheep, poultry, fish, crustaceans, etc.). As used herein, the term “patient” typically refers to a subject that is being treated for a disease or condition. The term “chimeric receptor” or “chimeric antigen receptor” as used herein refers to a polypeptide or a set of polypeptides, which when expressed in a cell, such as an immune effector cell, provides the cell with specificity for a target cell and the ability to negatively regulate intracellular signal transduction. DETAILED DESCRIPTION Provided herein are methods of preparing an immunotherapeutic with enhanced efficacy by treating lymphocytes with a B-cell lymphoma 2 (BCL-2) inhibitor, immunotherapeutic compositions produced by the methods herein, and methods of treating cancer therewith. Evading programed cell death, or apoptosis, is one of the many hallmarks of cancer. Several types of cancers have shown genetic mutations or protein overexpression of the anti- apoptotic BCL-2 family members that control cell survival and apoptosis. Therefore, small molecules, BH3 mimetics, targeting the BCL-2 family of proteins have become increasingly promising anti-cancer therapeutics when used alone or in combination with other chemotherapies. Venetoclax, the first FDA-approved BCL-2-specific BH3 mimetics, has accomplished unprecedent success as a monotherapy in treating patients with chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL) and in combination with chemotherapy in treating patients with acute myeloid leukemia (AML). Most of these studies focus on the direct apoptotic effect of venetoclax on cancers. However, little is known about the direct effect of venetoclax on other stromal and immune cells present on tumor microenvironment (TME) among them T cells. This is particularly relevant given that T cells are highly reliant on the BCL-2 family of proteins during stages of development, activation, contraction, and homeostasis. Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 Recent studies indicate that in mice venetoclax can enhance the antitumor efficacy of immune checkpoint blockade in a T cell manner. Another study illustrates that venetoclax can directly enhance the antileukemic activity of human T cells. BCL-2 inhibition by venetoclax during expansion of murine T cells results in an increased anti-apoptotic protein expression, cell death resistance, and alteration of anti-apoptotic dependency patterns while developing transcriptional reprogramming of naïve T cells towards gaining active like signature that characterized by upregulation of Jak-STAT and downregulation of MAPK and FOXO signaling pathways. Additionally, venetoclax induces BCL-2–dependent Treg to Th17 plasticity that enhances the antitumor efficacy of anti–PD-1 checkpoint blockade in a PI3K / AKT / FOXO dependent pathway. Mutations in BCL-2 have not only proven to cause cancer resistant to a wide array of conventional chemotherapies but also correlate with poor responses in patients treated with CD19CAR T cell therapy. Several studies have shown that the combination of CD19CAR T cells (CD19CARTs) with venetoclax can amplify the therapeutic outcomes in venetoclax-sensitive and resistant cancers. This combination aims to overcome the BCL-2 induced cancer relapse one of the significant barriers to the long-term therapeutic efficacy of CAR T cell therapy and suggest using CART overexpressing mutant BCL-2 to protect from any unwanted apoptotic effects of venetoclax on CARTs. Experiments conducted during development of embodiments herein reveal that pressuring the apoptotic system by venetoclax during the production and expansion of CARTs induces an adaptive-like reprogramming of T cells that resist cell death and exhaustion, improve the antitumor efficacy and metabolic fitness of CARTs in a non-apoptotic BCL-2 / STAT5 / AKT dependent manner. Further, the venetoclax-preconditioning approach can be beneficial to improve the therapeutic efficacy of CARTs prepared from patient apheresis product regardless the status of exhaustion or T cell composition of the patient’s PBMC sample. In experiments conducted during development of embodiments herein, human CD19CAR T cells (hCD19CARTs) were prepared and expanded in the presence of vehicle or venetoclax, followed by phenotypic and genetic analysis using flowcytometry and single cell sequencing (scRNA-seq). Ex vivo CART cell cytotoxicity and cytokine production following incubation with human diffuse large B cell lymphoma (DLBCL) was measured along with measurement of their in vivo efficacy in DLBCL xenograft murine models. Expanding hCD19CARTs in the Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 presence of venetoclax resulted in increased expression of anti-apoptotic proteins, upregulated transcription factors involved in T cell signaling and function and enhanced antitumor cytotoxicity with less exhaustion, higher cytokine and granzyme B production in vitro. Furthermore, venetoclax-treatedhCD19CARTs resulted in significantly improved tumor control and survival in pre-clinical xenograft models. scRNA-seq of venetoclax-treated hCD19CARTs before antigen stimulation revealed improved effector-like transcriptional profiles in various T cell compartments marked by enriched TNFα and IFNγ signatures and significant PI3K / AKT andSTAT5 signaling pathway enrichment and shifts towards oxidative phosphorylation metabolism, suggesting broad involvement of these pathways in the enhanced cytolytic function and survival of venetoclax-treated hCD19CARTs. Experiments conducted during development of embodiments herein functionally and mechanistically reveal that drugging BCL-2 in T cells with venetoclax, a BCL-2 specific BH3 mimetic, induces adaptive reprogramming profile that can be employed to benefit the fitness and therapeutic efficacy of CAR T cell products. Chimeric Antigen Receptor (CAR) T-cell therapy is a revolutionary immunotherapy approach that has shown great promise in treating various types of cancer. This therapy involves genetically engineering a patient's own T cells to express CARs on their surface, enabling them to target and destroy cancer cells more effectively. In some embodiments, the immune receptor is a chimeric immune receptor. In some embodiments, the immune receptor is a chimeric antigen receptor (CAR). In general, as used herein and unless otherwise specified, immune receptors in a CAR format refer to activating CARs that typically are a recombinant polypeptide construct comprising at least an extracellular antigen-binding domain, a transmembrane domain and a cytoplasmic signaling domain (also referred to herein as "an intracellular signaling domain") comprising a functional signaling domain derived from a stimulatory molecule. A CAR of the present disclosure may be a first, second, third, fourth, fifth, or subsequent generations of CARs. "First generation" CARs comprise a single intracellular signaling domain, generally derived from a T cell receptor chain. "First generation" CARs generally have the intracellular signaling domain from the CD3-zeta (CD3ζ) chain, which is the primary transmitter of signals from endogenous TCRs. "First generation" CARs can provide de novo antigen recognition and cause activation of both CD4+and CD8+T cells through their CD3ζ chain Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 signaling domain in a single fusion molecule, independent of HLA-mediated antigen presentation. "Second generation" CARs add a second intracellular signaling domain from one of various co-stimulatory molecules (e.g., CD28, 4-1BB, ICOS, OX40) to the cytoplasmic tail of the CAR to provide additional signals to the T cell. "Second generation" CARs provide both co- stimulation (e.g., CD28 or 4- 1BB) and activation (CD3ζ). Preclinical studies have indicated that "Second Generation" CARs can improve the anti-tumor activity of immunoresponsive cell, such as a T cell. "Third generation" CARs have multiple intracellular co-stimulation signaling domains (e.g., CD28 and 4-1BB) and an intracellular activation signaling domain (CD3ζ). “Fourth Generation” CARs are designed to secrete specific cytokines upon target engagement, enhancing the local immune response and potentially recruiting other immune cells. “Fifth Generation” CARs utilize membrane receptors. In some embodiments, the domains in the CAR polypeptide construct are in the same polypeptide chain, e.g., comprise a chimeric fusion protein. In some embodiments, the domains in the CAR polypeptide construct are not contiguous with each other, e.g., are in different polypeptide chains. In some embodiments, the stimulatory molecule is the zeta chain associated with the T cell receptor complex. In some embodiments, the cytoplasmic signaling domain comprises a primary signaling domain (e.g., a primary signaling domain of CD3-zeta). In some embodiments, the cytoplasmic signaling domain further comprises one or more functional signaling domains derived from at least one costimulatory molecule as defined below. In some embodiments, the costimulatory molecule is chosen from 4-1BB (i.e., CD 137), CD27, ICOS, and / or CD28. In some embodiments, the CAR. comprises a chimeric fusion protein comprising an extracellular antigen-binding domain, a transmembrane domain and an intracellular signaling domain comprising a functional signaling domain derived from a stimulatory molecule. In some embodiments, the CAR comprises a chimeric fusion protein comprising an extracellular antigen- binding domain, a transmembrane domain and an intracellular signaling domain comprising a functional signaling domain derived from a co-stimulatory molecule and a functional signaling domain derived from a stimulatory molecule. In some embodiments, the CAR comprises a chimeric fusion protein comprising an extracellular antigen-binding domain, a transmembrane domain and an intracellular signaling domain comprising two functional signaling domains derived from one or more co-stimulatory molecule(s) and a functional signaling domain derived from a stimulatory molecule. In some embodiments, the CAR comprises a chimeric fusion Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 protein comprising an extracellular antigen-binding domain, a transmembrane domain and an intracellular signaling domain comprising at least two functional signaling domains derived from one or more co-stimulatory molecule(s) and a functional signaling domain derived from a stimulatory molecule. In some embodiments, the CAR comprises an optional leader sequence (also referred to as a signal sequence) at the amino-terminus (N-term) of the CAR fusion protein. In some embodiments, the CAR further comprises a leader sequence at the N-terminus of the extracellular antigen-binding domain, wherein the leader sequence is optionally cleaved from the antigen recognition domain (e.g., an scFv) during cellular processing and localization of the CAR to the cellular membrane. Various chimeric antigen receptors are known in the art including, but not limited to, ScFv-FcεRIγCAIX, ScFv-FcεRIγ, ScFv-CD3ζ, ScFv-CD28-CD3ζ, ScFv-CD28-CD3ζ, ScFv - CD3ζ, ScFv-CD4-CD3ζ, CD3 ζ / CD137 / CD28, ScFv-CD28-41BB-CD3ζ, ScFv-CD8-CD3ζ, ScFv-FceRIγ, CD28 / 4-1BB-CD3ζ, ScFv-CD28mut-CD3ζ, Heregulin-CD3ζ, ScFv-CD28, ScFv- CD28-OX40-CD3ζ, ScFv-CD3ξ, IL-13-CD28-4-1BB-CD3ζ, IL-13-CD3ζ, IL-13-CD3ζ, ScFv- FcεRIγ, ScFV-CD4-FcεRIγ, ScFV-CD28-FcεRIγ, Ly49H-CD3ζ, NKG2D-CD3ζ, ScFv-b2c- CD3ζ, and FceRI-CD28-CD3ζ. In some embodiments, the chimeric antigen receptor has been modified to include control elements. In some embodiments, the chimeric antigen receptor is a split chimeric antigen receptor; see e.g., WO2017 / 091546. In some embodiments, the extracellular antigen recognition domain (e.g., ScFv) is capable of binding to a target selected from the group consisting of TSHR, CD19, CD123, CD22, BAFF-R, CD30, CD171, CS-1, CLL-1, CD33, EGFRvIII, GD2, GD3, BCMA, GPRC5D, Tn Ag, PSMA, ROR1, FLT3, FAP, TAG72, CD38, CD44v6, CEA, EPCAM, B7H3, KIT, IL- 13Ra2, mesothelin, IL-11Ra, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-0, SSEA-4, CD20, AFP, Folate receptor α, ERBB2 (Her2 / neu), MUC1, EGFR, CS1, CD138, NCAM, Claudin18.2, Prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gp100, bcr-ab1, tyrosinase, EphA2, Fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, Folate receptor R, TEM1 / CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD 179a, ALK, polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-1a, MAGE-A1, legumain, HPV E6 / E7, MAGE A1, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, prostate specific protein, survivin and telomerase, PCTA-1 / Galectin 8, Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 MelanA / MART1, Ras mutant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2ETS fusion gene), NA17, PAX3, androgen receptor, Cyclin B1, MYCN, RhoC, TRP- 2, CYP1B1, BORIS, SART3, PAX5, OY-TES 1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal tract carboxylesterase, mut hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, PD1, PDL1, PDL2, TGFβ, APRIL, and NKG2D. In some embodiments, the immune receptor is a chimeric TCR. A chimeric TCR generally includes an extracellular ligand binding domain grafted onto one or more constant domains of a TCR chain, for example a TCR alpha chain or TCR beta chain, to create a chimeric TCR that binds specifically to an antigen of interest, such a tumor-associated antigen. Without wishing to be bound by theory, it is believed that chimeric TCRs may signal through the TCR complex upon antigen binding. For example, an antibody or antibody fragment (e.g., scFv) can be grafted to the constant domain (e.g., at least a portion of the extracellular constant domain, the transmembrane domain and cytoplasmic domain) of a TCR chain, such as the TCR alpha chain and / or the TCR beta chain. As another example, the CDRs of an antibody or antibody fragment may be grafted into a TCR alpha chain and / or beta chain to create a chimeric TCR that binds specifically to an antigen. Such chimeric TCRs may be produced by methods known in the art (e.g., Willemsen RA et al., Gene Therapy 2000; 7:1369-1377; Zhang T et al., Cancer Gene Ther 200411: 487-496; and Aggen et al., Gene Ther. 2012 Apr; 19(4): 365-74; herein incorporated by reference for all purposes). The antigen of an immune receptor, such as a chimeric antigen receptor, can be a tumor- associated antigen. Immune receptors generally are capable of inducing signal transduction or changes in protein expression in the immune receptor-expressing cell that results in the modulation of an immune response upon binding to a cognate ligand (e.g., regulate, activate, initiate, stimulate, increase, prevent, attenuate, inhibit, reduce, decrease, inhibit, or suppress an immune response). For example, when CD3 chains present in a TCR / CAR cluster in response to ligand binding, an immunoreceptor tyrosine-based activation motifs (ITAMs)-meditated signal transduction cascade is produced. Specifically, in certain embodiments, when an endogenous TCR, exogenous TCR, chimeric TCR, or a CAR (specifically an activating CAR) binds their respective antigen, a formation of an immunological synapse occurs that includes clustering of Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 many molecules near the bound receptor (e.g. CD4 or CD8, CD3γ / δ / ε / ζ, etc.). This clustering of membrane bound signaling molecules allows for ITAM motifs contained within the CD3 chains to become phosphorylated that in turn can initiate a T cell activation pathway and ultimately activates transcription factors, such as NF-κΒ and AP-1. These transcription factors are capable of inducing global gene expression of the T cell to increase IL-2 production for proliferation and expression of master regulator T cell proteins in order to initiate a T cell mediated immune response, such as cytokine production and / or T cell mediated killing. Provided herein, in other aspects, are nucleic acids encoding at least one chimeric receptor as described above. In some embodiments, the nucleic acid encoding the at least one chimeric receptor is a vector. In some embodiments, the vector is selected from a plasmid vector, a viral vector, a lentiviral vector, or a phage vector. When the chimeric receptor is a multichain receptor, a set of polynucleotides is used. In this case, the set of polynucleotides can be cloned into a single vector or a plurality of vectors. In some embodiments, the polynucleotide comprises a sequence encoding a chimeric receptor, wherein the sequence encoding an extracellular ligand binding domain is contiguous with and in the same reading frame as a sequence encoding an intracellular signaling domain and a membrane localization domain. The polynucleotide can be codon optimized for expression in a mammalian cell. In some embodiments, the entire sequence of the polynucleotide has been codon optimized for expression in a mammalian cell. Codon optimization refers to the discovery that the frequency of occurrence of synonymous codons (i.e., codons that code for the same amino acid) in coding DNA is biased in different species. Such codon degeneracy allows an identical polypeptide to be encoded by a variety of nucleotide sequences. A variety of codon optimization methods is known in the art, and include, e.g., methods disclosed in at least US Patent Numbers 5,786,464 and 6,114,148, herein incorporated by reference for all purposes. The polynucleotide encoding a chimeric receptor can be obtained using recombinant methods known in the art, such as, for example by screening libraries from cells expressing the polynucleotide, by deriving it from a vector known to include the same, or by isolating directly from cells and tissues containing the same, using standard techniques. Alternatively, the polynucleotide can be produced synthetically, rather than cloned. Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 The polynucleotide can be cloned into a vector. In some embodiments, an expression vector known in the art is used. Accordingly, the present disclosure includes retroviral and lentiviral vector constructs expressing a chimeric receptor that can be directly transduced into a cell. The present disclosure also includes an RNA construct that can be directly transfected into a cell. A method for generating mRNA for use in transfection involves in vitro transcription (IVT) of a template with specially designed primers, followed by polyA addition, to produce a construct containing 3’ and 5’ untranslated sequence (“UTR”) (e.g., a 3’ and / or 5’ UTR described herein), a 5’ cap (e.g., a 5’ cap described herein) and / or Internal Ribosome Entry Site (IRES) (e.g., an IRES described herein), the nucleic acid to be expressed, and a polyA tail. RNA so produced can efficiently transfect different kinds of cells. In some embodiments, an RNA chimeric receptor vector is transduced into a cell, e.g., a T cell or a NK cell, by electroporation. In some embodiments, a vector of the present disclosure may further comprise a signal sequence to facilitate secretion, a polyadenylation signal and transcription terminator, an element allowing episomal replication, and / or elements allowing for selection. Engineered Cells Also provided herein are genetically engineered cells comprising a nucleic acid encoding at least one chimeric receptor of the present disclosure or that express a chimeric receptor of the present disclosure. Various ways of introducing nucleic acids / vectors (i.e., genetically engineering) are known to those having skill in the art and include, are not limited to, transduction (i.e., viral infection), transformation, and transfection. Mechanisms of transfection include chemical-based transfection (e.g., calcium phosphate-mediated, lipofection / liposome mediated, etc.), non-chemical-based transfection (e.g., electroporation, cell squeezing, sonoporation, optical transfection, protoplast fusion, impalefection, hydrodynamic delivery, etc.), and particle-based transfection (e.g., gene gun, magnetofection, particle bombardment, etc.). In some embodiments, a genetically engineered cell of the present disclosure is an immunomodulatory cell. Immunomodulatory cells include, but are not limited to, a T cell, a CD8+ T cell, a CD4+ T cell, a gamma-delta T cell, a cytotoxic T lymphocyte (CTL), a regulatory T cell, a viral-specific T cell, a Natural Killer T (NKT) cell, a Natural Killer (NK) cell, a B cell, a tumor-infiltrating lymphocyte (TIL), an innate lymphoid cell, a mast cell, an Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 eosinophil, a basophil, a neutrophil, a myeloid cell, a macrophage, a monocyte, a dendritic cell, an ESC-derived cell, and an iPSC-derived cell. In some embodiments, a genetically engineered cell of the present disclosure is an immune cell. In some embodiments, the immune cell is a T cell. Examples of T cells include, but are not limited to CD8+ T cells, CD4+ T cells, effector cells, helper cells (TH cells), cytotoxic cells (TC cells, CTLs, T-killer cells, killer T cells), memory cells (central memory T cells, effector memory T cells, tissue resident memory T cells, virtual memory T cells, etc.), regulatory T cells (e.g., CD4+, FOXP3+, CD25+), natural killer T cells, mucosal associated invariant cells, and gamma delta T cells. In some embodiments, a genetically engineered cell is autologous. In some embodiments, a genetically engineered cell is allogeneic. In some embodiments, the genetically engineered cell is further engineered to express an exogenous immune receptor. For example, the genetically engineered cell can be engineered to express a chimeric immune receptor, such as a CAR. In another example, the genetically engineered cell can be engineered to express a naturally-occurring immune receptor exogenous to the engineered cell. In some embodiments, the genetically engineered cell is engineered to express a chimeric receptor and an exogenous immune receptor. The genetically engineered cell can be engineered to express both a chimeric receptor and an exogenous immune receptor simultaneously (e.g., polynucleotides encoding each receptor are introduced simultaneously). The genetically engineered cell can be engineered to express both a chimeric receptor and an exogenous immune receptor sequentially (e.g., first engineered to express either the chimeric receptor and the exogenous immune receptor, then subsequently engineered to express the other receptor). In another aspect, the present disclosure provides a method of preparing a genetically engineered cell (e.g., a genetically engineered immunomodulatory cell) expressing or capable of expressing a chimeric receptor for experimental or therapeutic use. In another aspect, the present disclosure provides a method of preparing a genetically engineered cell (e.g., a genetically engineered immunomodulatory cell) expressing or capable of expressing a chimeric receptor and an immune receptor for experimental or therapeutic use. Ex vivo procedures for making therapeutic chimeric receptor- engineered cells are well known in the art. For example, cells are isolated from a mammal (e.g., a human) and genetically Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 engineered (i.e., transduced or transfected in vitro) with a vector expressing a chimeric receptor disclosed herein. The chimeric receptor- engineered cell can be administered to a mammalian recipient to provide a therapeutic benefit. The mammalian recipient may be a human and the chimeric receptor-modified cell can be autologous with respect to the recipient. Alternatively, the cells can be allogeneic, syngeneic or xenogeneic with respect to the recipient. In some embodiments, the population of cells is cultured for a period of time that results in the production of an expanded cell population that comprises at least 2-fold the number of cells of the starting population. In some embodiments, the population of cells is cultured for a period of time that results in the production of an expanded cell population that comprises at least 4-fold the number of cells of the starting population. In some embodiments, the population of cells is cultured for a period of time that results in the production of an expanded cell population that comprises at least 16-fold the number of cells of the starting population. In some embodiments, methods described herein employ a BCL-2 inhibitor or a composition / method for the inhibition of BCL-2 activity or expression. In some embodiments, compositions are provided comprising CAR lymphocytes that have been enhanced through the inhibition of BCL-2. In some embodiments, provided herein are inhibitors of BCL-2 activity. In some embodiments, an inhibitor is a small molecule, peptide, antibody, antibody fragment, a genome editing agent, etc. that upon administration to a cell, reduces the activity of BCL-2. In some embodiments, a BCL-2 inhibitor is an antibody or antibody fragment. In some embodiments, a BCL-2 inhibitor is an antibody or antibody fragment that binds to BCL-2 and reduces BCL-2 activity. In some embodiments, an anti-BCL-2 antibody or antibody fragment is provided that prevents or reduces binding of BCL-2 to a ligand thereof. In some embodiments, a BCL-2 inhibitor is a BH3 mimetic. In some embodiments, a BH3 mimetic is selected from venetoclax, A1155463, A-1331852, S63845, AMG-176, S55746, ABT-199 (4-[4-[[2-(4-Chlorophenyl)-4,4-dimethylcyclohex-1-en-1-yl]methyl]piperazin-1-yl]-N- [[3-nitro-4-[[(tetrahydro-2H-pyran-4-yl)methyl]amino]phenyl]sulfonyl]-2-[(1H-pyrrolo[2,3- b]pyridin-5-yl)oxy]benzamide), ABT-737 (4-[4-[[2-(4-chlorophenyl)phenyl]methyl]piperazin-1- yl]-N-[4-[[(2R)-4-(dimethylamino)-1-phenylsulfanylbutan-2-yl] amino]-3- nitrophenyl]sulfonylbenzamide) (navitoclax), ABT-263 ((R)-4-(4-((4′-chloro-4,4-dimethyl- 3,4,5,6-tetrahydro-[1,1′-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((4-((4-morpholino-1- Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 (phenylthio)butan-2-yl)amino)-3((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide), GX15- 070 (obatoclax mesylate, (2Z)-2-[(5Z)-5-[(3,5-dimethyl-1H-pyrrol-2-yl)methylidene]-4- methoxypyrrol-2-ylidene]indole; methanesulfonic acid))), 2-methoxy-antimycin A3, YC137 (4- (4,9-dioxo-4,9-dihydronaphtho[2,3-d]thiazol-2-ylamino)-phenyl ester), pogosin, ethyl 2-amino- 6-bromo-4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate, Nilotinib-d3, TW-37 (N-[4-[[2-(1,1-Dimethylethyl)phenyl]sulfonyl]phenyl]-2,3,4-trihydroxy-5-[[2-(1- methylethyl)phenyl]methyl]benzamide), Apogossypolone (ApoG2), HA14-1, AT101 (gossypol), sabutoclax, gambogic acid, and G3139 (Oblimersen). In some embodiments, provided herein are inhibitors of BCL-2 expression. In some embodiments, an inhibitor is a small molecule, peptide, antibody, antibody fragment, a genome editing agent, etc. In particular embodiments, a BCL-2 inhibitor is a nucleic acid-based inhibitor. In some embodiments, the inhibitor is a small molecule, an aptamer, a siRNA, a shRNA, a miRNA, a morpholino, a ribozyme, an antisense nucleic acid molecule, a CRISPR-Cas9-based construct, a CRISPR-Cpf1-based construct, a meganuclease, a zinc finger nuclease, a transcription activator-like (TAL) effector (TALE) nuclease, etc. In some embodiments, the BCL-2 inhibitor is a small interfering RNA (siRNA), also known as short interfering RNA or silencing RNA. In some embodiments, an siRNA is an 18 to 30 nucleotide, preferably 19 to 25 nucleotide, most preferred 21 to 23 nucleotide or even more preferably 21 nucleotide-long double-stranded RNA molecule. siRNA is involved in the RNA interference (RNAi) pathway where the siRNA interferes with the expression of a specific gene (e.g., the BCL-2 ). siRNAs naturally found in nature have a well-defined structure: a short double-strand of RNA (dsRNA) with 2-nt 3' overhangs on either end. Each strand has a 5' phosphate group and a 3' hydroxyl (--OH) group. This structure is the result of processing by dicer, an enzyme that converts either long dsRNAs or small hairpin RNAs into siRNAs. siRNAs can also be exogenously (artificially) introduced into cells to bring about the specific knockdown of a gene of interest (e.g., the BCL-2 ). Essentially any gene for which the sequence is known can thus be targeted based on sequence complementarity with an appropriately tailored siRNA. The double-stranded RNA molecule or a metabolic processing product thereof is capable of mediating target-specific nucleic acid modifications, particularly RNA interference and / or DNA methylation. Exogenously introduced siRNAs may be devoid of overhangs at their 3' and 5' ends, however, in some embodiments at least one RNA strand has a 5'- and / or 3'-overhang. Preferably, Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 one end of the double-strand has a 3'-overhang from 1 to 5 nucleotides, more preferably from 1 to 3 nucleotides and most preferably 2 nucleotides. The other end may be blunt-ended or has up to 6 nucleotides 3'-overhang. In general, any RNA molecule suitable to act as siRNA and inhibit the BCL-2 is envisioned in the present invention. In some embodiments, siRNA duplexes are provided composed of 21-nt sense and 21-nt antisense strands, paired in a manner to have a 2-nt 3'-overhang. The sequence of the 2-nt 3' overhang makes a small contribution to the specificity of target recognition restricted to the unpaired nucleotide adjacent to the first base pair. 2'- deoxynucleotides in the 3' overhangs are as efficient as ribonucleotides, but are often cheaper to synthesize and probably more nuclease resistant. Delivery of siRNA may be accomplished using any of the methods known in the art, for example by combining the siRNA with saline and administering the combination intravenously or intranasally or by formulating siRNA in glucose (such as for example 5% glucose) or cationic lipids and polymers can be used for siRNA delivery in vivo through systemic routes either intravenously (IV) or intraperitoneally (IP). In some embodiments, provided herein are siRNA molecules that target and inhibit the expression (e.g., knock down) of the BCL-2 A short hairpin RNA (shRNA) is a sequence of RNA that makes a tight hairpin turn that can be used to silence gene expression (e.g., of BCL-2) via RNA interference. In some embodiments, shRNA uses a vector introduced into cells and utilizes the U6 promoter to ensure that the shRNA is always expressed. This vector is usually passed on to daughter cells, allowing the gene silencing to be inherited. The shRNA hairpin structure is cleaved by the cellular machinery into siRNA, which is then bound to the RNA-induced silencing complex (RISC). The RISC binds to and cleaves RNAs which match the siRNA that is bound to (e.g., comprising the sequence of the BCL-2 ). In some embodiments, si / shRNAs to be used in the present invention are chemically synthesized using appropriately protected ribonucleoside phosphoramidites and a conventional DNA / RNA synthesizer. In some embodiments, provided herein are shRNA molecules that target and inhibit the expression (e.g., knock down) of the BCL-2. Further molecules effecting RNAi (and useful herein for the inhibition of expression of the BCL-2 ) include, for example, microRNAs (miRNA). Said RNA species are single-stranded RNA molecules. Endogenously present miRNA molecules regulate gene expression by binding to a complementary mRNA transcript and triggering of the degradation of said mRNA transcript through a process similar to RNA interference. Accordingly, exogenous miRNA may be Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 employed as an inhibitor of the BCL-2 after introduction into target cells. In some embodiments, provided herein are miRNA molecules that target and inhibit the expression (e.g., knock down) of the BCL-2 . Morpholinos (or morpholino oligonucleotides) are synthetic nucleic acid molecules having a length of about 20 to 30 nucleotides and, typically about 25 nucleotides. Morpholinos bind to complementary sequences of target transcripts (e.g., the BCL-2 ) by standard nucleic acid base-pairing. They have standard nucleic acid bases which are bound to morpholine rings instead of deoxyribose rings and linked through phosphorodiamidate groups instead of phosphates. Due to replacement of anionic phosphates into the uncharged phosphorodiamidate groups, ionization in the usual physiological pH range is prevented, so that morpholinos in organisms or cells are uncharged molecules. The entire backbone of a morpholino is made from these modified subunits. Unlike inhibitory small RNA molecules, morpholinos do not degrade their target RNA molecules. Rather, they sterically block binding to a target sequence within a RNA and prevent access by molecules that might otherwise interact with the RNA. In some embodiments, provided herein are morpholino oligonucleotides that target and inhibit the expression (e.g., knock down) of the BCL-2 . A ribozyme (ribonucleic acid enzyme, also called RNA enzyme or catalytic RNA) is an RNA molecule that catalyzes a chemical reaction. Many natural ribozymes catalyze either their own cleavage or the cleavage of other RNAs, but they have also been found to catalyze the aminotransferase activity of the ribosome. Non-limiting examples of well-characterized small self-cleaving RNAs are the hammerhead, hairpin, hepatitis delta virus, and in vitro-selected lead- dependent ribozymes, whereas the group I intron is an example for larger ribozymes. The principle of catalytic self-cleavage is well established. Since it was shown that hammerhead structures can be integrated into heterologous RNA sequences and that ribozyme activity can thereby be transferred to these molecules, catalytic antisense sequences can be engineered for almost any target sequence can be created, provided the target sequence contains a potential matching cleavage site. The basic principle of constructing hammerhead ribozymes is as follows: A region of interest of the RNA (e.g., a portion of the BCL-2 ), which contains the GUC (or CUC) triplet, is selected. Two oligonucleotide strands, each usually with 6 to 8 nucleotides, are taken and the catalytic hammerhead sequence is inserted between them. In some embodiments, provided herein are ribozyme inhibitors oligonucleotides of the BCL-2. Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 In some embodiments, BCL-2 is inhibited (and / or BCL-2 activity is inhibited) by modifying the BCL-2 sequence in target cells. In some embodiments, the alteration of the BCL-2 is carried out using one or more DNA-binding nucleic acids, such as alteration via an RNA- guided endonuclease (RGEN). For example, the alteration can be carried out using clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins. In general, "CRISPR system" refers collectively to transcripts and other elements involved in the expression of or directing the activity of CRISPR-associated ("Cas") genes, including sequences encoding a Cas gene, a tracr (trans-activating CRISPR) sequence (e.g. tracrRNA or an active partial tracrRNA), a tracr-mate sequence (encompassing a "direct repeat" and a tracrRNA-processed partial direct repeat in the context of an endogenous CRISPR system), a guide sequence (also referred to as a "spacer" in the context of an endogenous CRISPR system), and / or other sequences and transcripts from a CRISPR locus. The CRISPR / Cas nuclease or CRISPR / Cas nuclease system can include a non-coding RNA molecule (guide) RNA, which sequence-specifically binds to DNA, and a Cas protein (e.g., Cas9), with nuclease functionality (e.g., two nuclease domains). One or more elements of a CRISPR system can derive from a type I, type II, or type III CRISPR system, e.g., derived from a particular organism comprising an endogenous CRISPR system, such as Streptococcus pyogenes. In some aspects, a Cas nuclease and gRNA (including a fusion of crRNA specific for the target sequence (e.g., a sequence within the BCL-2 ) and fixed tracrRNA) are introduced into the cell. In general, target sites at the 5' end of the gRNA target the Cas nuclease to the target site, e.g., the BCL-2 , using complementary base pairing. The target site may be selected based on its location immediately 5' of a protospacer adjacent motif (PAM) sequence, such as typically NGG, or NAG. In this respect, the gRNA is targeted to the desired sequence by modifying the first 20, 19, 18, 17, 16, 15, 14, 14, 12, 11, or 10 nucleotides of the guide RNA to correspond to the target DNA sequence (e.g., sequence within the BCL-2 ). In general, a CRISPR system is characterized by elements that promote the formation of a CRISPR complex at the site of a target sequence. Typically, "target sequence" generally refers to a sequence to which a guide sequence is designed to have complementarity, where hybridization between the target sequence and a guide sequence promotes the formation of a CRISPR complex. Full complementarity is not necessarily required, provided there is sufficient complementarity to cause hybridization and promote formation of a CRISPR complex. The CRISPR system can induce double stranded Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 breaks (DSBs) at the SRC-3 target site, followed by disruptions or alterations as discussed herein. In other embodiments, Cas9 variants, deemed "nickases," are used to nick a single strand at the target site (e.g., within the BCL-2 ). Paired nickases can be used, e.g., to improve specificity, each directed by a pair of different gRNAs targeting sequences such that upon introduction of the nicks simultaneously, a 5' overhang is introduced. In other embodiments, catalytically inactive Cas9 is fused to a heterologous effector domain such as a transcriptional repressor or activator, to affect gene expression (e.g., to inhibit expression of the BCL-2 ). In some embodiments, the CRISPR system is used to alter the BCL-2 , inhibit expression of the BCL-2 , and / or to inactivate the expression product of the BCL-2 . The term "antisense nucleic acid molecule" or “antisense oligonucleotide” as used herein, refers to a nucleic acid which is complementary to a target nucleic acid. An antisense molecule in accordance with the invention is capable of interacting with the target nucleic acid, more specifically it is capable of hybridizing with the target nucleic acid. Due to the formation of the hybrid, transcription of the target gene(s) and / or translation of the target mRNA is reduced or blocked. Standard methods relating to antisense technology have been described (see, e.g., Melani et al., Cancer Res. (1991) 51:2897-2901). In some embodiments, provided herein are antisense oligonucleotides capable of inhibiting expression of BCL-2when administered to cell or subject. In some embodiments, the antisense oligonucleotides are antisense DNA- and / or RNA-oligonucleotides. In some embodiments, provided herein are modified antisense oligonucleotides, such as, antisense 2'-O-methyl oligo-ribonucleotides, antisense oligonucleotides containing phosphorothiaote linkages, antisense oligonucleotides containing Locked Nucleic Acid LNA(R) bases, morpholino antisense oligonucleotides, PPAR-gamma agonists, antagomirs. In some embodiments, ASOs comprise Locked Nucleic Acid (LNA) or 2’- methoxyethyl (MOE) modifications (inter-nucleotide linkages are phosphorothioates interspersed with phosphodiesters, and all cytosine residues are 5’-methylcytosines). In some embodiments, genetically engineered cells (e.g., CAR T cells), having been prepared and / or enhanced by the methods herein (e.g., with BCL-2 inhibition) can be formulated in pharmaceutical compositions. Pharmaceutical compositions of the present disclosure can comprise genetically engineered cells (e.g., CAR T cells), as described herein, in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents or excipients. Such materials should be non-toxic and should not interfere with the efficacy of the active Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 ingredient. The precise nature of the carrier or other material can depend on the route of administration, e.g. oral, intravenous, cutaneous or subcutaneous, nasal, intramuscular, intraperitoneal routes. In certain embodiments, the composition is directly injected into an organ of interest (e.g., an organ affected by a disorder). Alternatively, the composition may be provided indirectly to the organ of interest, for example, by administration into the circulatory system. Expansion and differentiation agents can be provided prior to, during, or after administration of the composition to increase production of T cells in vitro or in vivo. In certain embodiments, the compositions are pharmaceutical compositions comprising genetically engineered cells, such as immunomodulatory or immune cells, or their progenitors and a pharmaceutically acceptable carrier. Administration can be autologous or heterologous. For example, immunomodulatory or immune cells, or progenitors, can be obtained from one subject, and administered to the same subject or a different, compatible subject. In some embodiments, genetically engineered cells, such as immunomodulatory or immune cells, or their progeny may be derived from peripheral blood cells (e.g., in vivo, ex vivo, or in vitro derived) and may be administered via localized injection, including catheter administration, systemic injection, localized injection, intravenous injection, or parenteral administration. When administering a therapeutic composition of the present disclosure (e.g., a pharmaceutical composition containing a genetically engineered cell of the present disclosure), it will generally be formulated in a unit dosage injectable form (solution, suspension, emulsion). Certain aspects of the present disclosure relate to formulations of compositions comprising chimeric receptors of the present disclosure or genetically engineered cells (e.g., immunomodulatory or immune cells of the present disclosure) expressing such chimeric receptors. In some embodiments, compositions of the present disclosure comprising genetically engineered cells may be provided as sterile liquid preparations, including without limitation isotonic aqueous solutions, suspensions, emulsions, dispersions, and viscous compositions, which may be buffered to a selected pH. Liquid preparations are typically easier to prepare than gels, other viscous compositions, and solid compositions. Additionally, liquid compositions may be more convenient to administer, especially by injection. In some embodiments, viscous compositions can be formulated within the appropriate viscosity range to provide longer contact periods with specific tissues. Liquid or viscous compositions can comprise carriers, which can be a solvent or dispersing medium containing, for example, water, saline, phosphate buffered saline, Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 polyol (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.) and suitable mixtures thereof. Pharmaceutical compositions for oral administration can be in tablet, capsule, powder or liquid form. A tablet can include a solid carrier such as gelatin or an adjuvant. Liquid pharmaceutical compositions generally include a liquid carrier such as water, petroleum, animal or vegetable oils, mineral oil or synthetic oil. Physiological saline solution, dextrose or other saccharide solution or glycols such as ethylene glycol, propylene glycol or polyethylene glycol can be included. For intravenous, cutaneous or subcutaneous injection, or injection at the site of affliction, the active ingredient will be in the form of a parenterally acceptable aqueous solution which is pyrogen-free and has suitable pH, isotonicity and stability. Those of relevant skill in the art are well able to prepare suitable solutions using, for example, isotonic vehicles such as Sodium Chloride Injection, Ringer's Injection, Lactated Ringer's Injection. Preservatives, stabilizers, buffers, antioxidants and / or other additives can be included, as required. In some embodiments, compositions of the present disclosure can be isotonic, i.e., having the same osmotic pressure as blood and lacrimal fluid. In some embodiments, the desired isotonicity may be achieved using, for example, sodium chloride, dextrose, boric acid, sodium tartrate, propylene glycol, or other inorganic or organic solutes. In some embodiments, compositions of the present disclosure may further include various additives that may enhance the stability and sterility of the compositions. Examples of such additives include, without limitation, antimicrobial preservatives, antioxidants, chelating agents, and buffers. In some embodiments, microbial contamination may be prevented by the inclusions of any of various antibacterial and antifungal agents, including without limitation parabens, chlorobutanol, phenol, sorbic acid, and the like. Prolonged absorption of an injectable pharmaceutical formulation of the ;present disclosure can be brought about by the use of suitable agents that delay absorption, such as aluminum monostearate and gelatin. In some embodiments, sterile injectable solutions can be prepared by incorporating genetically modified cells of the present disclosure in a sufficient amount of the appropriate solvent with various amounts of any other ingredients, as desired. Such compositions may be in admixture with a suitable carrier, diluent, or excipient such as sterile water, physiological saline, glucose, dextrose, or the like. In Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 some embodiments, the compositions can also be lyophilized. The compositions can contain auxiliary substances such as wetting, dispersing agents, pH buffering agents, and antimicrobials depending upon the route of administration and the preparation desired. In some embodiments, the components of the formulations of the present disclosure are selected to be chemically inert and to not affect the viability or efficacy of the genetically modified cells of the present disclosure. One consideration concerning the therapeutic use of the genetically engineered cells of the present disclosure is the quantity of cells needed to achieve optimal efficacy. In some embodiments, the quantity of cells to be administered will vary for the subject being treated. In certain embodiments, the quantity of genetically engineered cells that are administered to a subject in need thereof may range from 1 x 104cells to 1 x 1010cells. In some embodiments, the precise quantity of cells that would be considered an effective dose may be based on factors individual to each subject, including their size, age, sex, weight, and condition of the particular subject. Dosages can be readily ascertained by those skilled in the art based on the present disclosure and the knowledge in the art. A composition can be administered alone or in combination with other treatments, either simultaneously or sequentially dependent upon the condition to be treated. In some embodiments, the immunotherapeutics with enhanced efficacy described herein are administered to a subject for the treatment of a cancer. Certain embodiments herein are directed to administration of immunotherapeutics with enhanced efficacy described herein to a subject with cancer, in remission from cancer, or at elevated risk of cancer. In some embodiments, immunotherapeutics with enhanced efficacy described herein are administered as part of therapeutic or prophylactic regimen for the treatment or prevention of acute myeloid leukemia, cancer in adolescents, adrenocortical carcinoma childhood, AIDS-related cancers (e.g., Lymphoma and Kaposi's Sarcoma), anal cancer, appendix cancer, astrocytomas, atypical teratoid, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem glioma, brain tumor, bronchial tumors, burkitt lymphoma, carcinoid tumor, atypical teratoid, embryonal tumors, germ cell tumor, primary lymphoma, cervical cancer, childhood cancers, chordoma, cardiac tumors, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myleoproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), embryonal tumors, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, esthesioneuroblastoma, ewing sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, fibrous histiocytoma of bone, gall bladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumors (GIST), germ cell tumor, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumors, pancreatic neuroendocrine tumors, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic squamous neck cancer with occult primary, midline tract carcinoma, mouth cancer multiple endocrine neoplasia syndromes, multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplastic syndromes, myelodysplastic / myeloproliferative neoplasms, multiple myeloma, merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma of bone and osteosarcoma, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-hodgkin lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pleuropulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, stomach (gastric) cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T- Cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell cancer of the renal pelvis and ureter, trophoblastic tumor, unusual cancers of childhood, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or viral-induced cancer. In some embodiments, immunotherapeutics with enhanced efficacy described herein are co-administered along with administration of a chemotherapy agent. In some embodiments, the chemotherapeutic is selected from the group consisting of mitotic inhibitors, alkylating agents, anti-metabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzyme inhibitors, topoisomerase inhibitors, protein-protein interaction inhibitors, biological response modifiers, anti-hormones, angiogenesis inhibitors, and anti-androgens. Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 Non-limiting examples are chemotherapeutic agents, cytotoxic agents, and non-peptide small molecules such as Gleevec® (Imatinib Mesylate), Velcade® (bortezomib), Casodex (bicalutamide), Iressa® (gefitinib), and Adriamycin as well as a host of chemotherapeutic agents. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide (CYTOXANTM); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethylenethiophosphaoramide and trimethylolomelamine; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, CasodexTM, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5- oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6- mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6- azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllinic acid; 2- ethylhydrazide; procarbazine; PSK.RTM.; razoxane; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2''-trichlorotriethylamine; urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxanes, e.g., paclitaxel (TAXOLTM, Bristol-Myers Squibb Oncology, Princeton, N.J.) Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 and docetaxel (TAXOTERETM, Rhone-Poulenc Rorer, Antony, France); retinoic acid; esperamicins; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the above. Also included as suitable chemotherapeutic cell conditioners are anti-hormonal agents that act to regulate or inhibit hormone action on tumors such as anti-estrogens including for example tamoxifen, (NolvadexTM), raloxifene, aromatase inhibiting 4(5)-imidazoles, 4- hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone, and toremifene (Fareston); and anti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; camptothecin-11 (CPT-11); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO). Where desired, the compounds or pharmaceutical composition of the present invention can be used in combination with commonly prescribed anti- cancer drugs such as Herceptin®, Avastin®, Erbitux®, Rituxan®, Taxol®, Arimidex®, Taxotere®, ABVD, AVICINE, Abagovomab, Acridine carboxamide, Adecatumumab, 17-N- Allylamino-17-demethoxygeldanamycin, Alpharadin, Alvocidib, 3-Aminopyridine-2- carboxaldehyde thiosemicarbazone, Amonafide, Anthracenedione, Anti-CD22 immunotoxins, Antineoplastic, Antitumorigenic herbs, Apaziquone, Atiprimod, Azathioprine, Belotecan, Bendamustine, BIBW 2992, Biricodar, Brostallicin, Bryostatin, Buthionine sulfoximine, CBV (chemotherapy), Calyculin, cell-cycle nonspecific antineoplastic agents, Dichloroacetic acid, Discodermolide, Elsamitrucin, Enocitabine, Epothilone, Eribulin, Everolimus, Exatecan, Exisulind, Ferruginol, Forodesine, Fosfestrol, ICE chemotherapy regimen, IT-101, Imexon, Imiquimod, Indolocarbazole, Irofulven, Laniquidar, Larotaxel, Lenalidomide, Lucanthone, Lurtotecan, Mafosfamide, Mitozolomide, Nafoxidine, Nedaplatin, Olaparib, Ortataxel, PAC-1, Pawpaw, Pixantrone, Proteasome inhibitor, Rebeccamycin, Resiquimod, Rubitecan, SN-38, Salinosporamide A, Sapacitabine, Stanford V, Swainsonine, Talaporfin, Tariquidar, Tegafur- uracil, Temodar, Tesetaxel, Triplatin tetranitrate, Tris(2-chloroethyl)amine, Troxacitabine, Uramustine, Vadimezan, Vinflunine, ZD6126 or Zosuquidar. Embodiments herein further relate to methods for using a immunotherapeutics with enhanced efficacy described herein in combination with radiation therapy for inhibiting abnormal cell growth or treating the hyperproliferative disorder in the mammal. Techniques for Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 administering radiation therapy are known in the art, and these techniques can be used in the combination therapy described herein. The administration of immunotherapeutics with enhanced efficacy described herein in this combination therapy can be determined as described herein. Radiation therapy can be administered through one of several methods, or a combination of methods, including without limitation external-beam therapy, internal radiation therapy, implant radiation, stereotactic radiosurgery, systemic radiation therapy, radiotherapy and permanent or temporary interstitial brachytherapy. The term “brachytherapy,” as used herein, refers to radiation therapy delivered by a spatially confined radioactive material inserted into the body at or near a tumor or other proliferative tissue disease site. The term is intended without limitation to include exposure to radioactive isotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and radioactive isotopes of Lu). Suitable radiation sources for use as a cell conditioner of the present invention include both solids and liquids. By way of non-limiting example, the radiation source can be a radionuclide, such as I-125, I-131, Yb-169, Ir-192 as a solid source, I-125 as a solid source, or other radionuclides that emit photons, beta particles, gamma radiation, or other therapeutic rays. The radioactive material can also be a fluid made from any solution of radionuclide(s), e.g., a solution of I-125 or I-131, or a radioactive fluid can be produced using a slurry of a suitable fluid containing small particles of solid radionuclides, such as Au-198, Y-90. Moreover, the radionuclide(s) can be embodied in a gel or radioactive micro spheres. Immunotherapeutics with enhanced efficacy described herein may also be used in combination with an amount of one or more substances selected from anti-angiogenesis agents, signal transduction inhibitors, antiproliferative agents, glycolysis inhibitors, or autophagy inhibitors. Anti-angiogenesis agents, such as MMP-2 (matrix-metalloproteinase 2) inhibitors, MMP- 9 (matrix-metalloprotienase 9) inhibitors, and COX-11 (cyclooxygenase 11) inhibitors, can be used in conjunction with immunotherapeutics with enhanced efficacy described herein. Anti- angiogenesis agents include, for example, rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II inhibitors include CELEBREXTM (alecoxib), valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in WO 96 / 33172 (published October 24,1996), WO 96 / 27583 (published March 7,1996), European Patent Application No. 97304971.1 (filed July Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 8,1997), European Patent Application No. 99308617.2 (filed October 29, 1999), WO 98 / 07697 (published February 26,1998), WO 98 / 03516 (published January 29,1998), WO 98 / 34918 (published August 13,1998), WO 98 / 34915 (published August 13,1998), WO 98 / 33768 (published August 6,1998), WO 98 / 30566 (published July 16, 1998), European Patent Publication 606,046 (published July 13,1994), European Patent Publication 931, 788 (published July 28,1999), WO 90 / 05719 (published May 31,1990), WO 99 / 52910 (published October 21,1999), WO 99 / 52889 (published October 21, 1999), WO 99 / 29667 (published June 17,1999), PCT International Application No. PCT / IB98 / 01113 (filed July 21,1998), European Patent Application No. 99302232.1 (filed March 25,1999), Great Britain Patent Application No. 9912961.1 (filed June 3, 1999), United States Provisional Application No. 60 / 148,464 (filed August 12,1999), United States Patent 5,863, 949 (issued January 26,1999), United States Patent 5,861, 510 (issued January 19,1999), and European Patent Publication 780,386 (published June 25, 1997), all of which are incorporated herein in their entireties by reference. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity inhibiting MMP-1. More preferred, are those that selectively inhibit MMP-2 and / or AMP-9 relative to the other matrix- metalloproteinases (e.g., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP- 7, MMP-8, MMP- 10, MMP-ll, MMP-12, andMMP-13). Some specific examples of MMP inhibitors useful in the invention are AG-3340, RO 32-3555, and RS 13-0830. Autophagy inhibitors include, but are not limited to chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil™), bafilomycin A1, 5-amino-4-imidazole carboxamide riboside (AICAR), okadaic acid, autophagy-suppressive algal toxins which inhibit protein phosphatases of type 2A or type 1, analogues of cAMP, and drugs which elevate cAMP levels such as adenosine, LY204002, N6-mercaptopurine riboside, and vinblastine. In addition, antisense or siRNA that inhibits expression of proteins including but not limited to ATG5 (which are implicated in autophagy), may also be used. In some embodiments, medicaments which are administered in conjunction with immunotherapeutics with enhanced efficacy described herein include any suitable drugs usefully delivered by inhalation for example, analgesics, e.g., codeine, dihydromorphine, ergotamine, fentanyl or morphine; anginal preparations, e.g., diltiazem; antiallergics, e.g., cromoglycate, ketotifen or nedocromil; anti-infectives, e.g., cephalosporins, penicillins, streptomycin, sulphonamides, tetracyclines or pentamidine; antihistamines, e.g., methapyrilene; anti- Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 inflammatories, e.g., beclomethasone, flunisolide, budesonide, tipredane, triamcinolone acetonide or fluticasone; antitussives, e.g., noscapine; bronchodilators, e.g., ephedrine, adrenaline, fenoterol, formoterol, isoprenaline, metaproterenol, phenylephrine, phenylpropanolamine, pirbuterol, reproterol, rimiterol, salbutamol, salmeterol, terbutalin, isoetharine, tulobuterol, orciprenaline or (-)-4-amino-3,5-dichloro-α-[[[6-[2-(2- pyridinyl)ethoxy]hexyl]-amino]methyl]benzenemethanol; diuretics, e.g., amiloride; anticholinergics e.g., ipratropium, atropine or oxitropium; hormones, e.g., cortisone, hydrocortisone or prednisolone; xanthines e.g., aminophylline, choline theophyllinate, lysine theophyllinate or theophylline; and therapeutic proteins and peptides, e.g., insulin or glucagon Exemplary therapeutic agents useful for a combination therapy with immunotherapeutics with enhanced efficacy described herein include but are not limited to agents as described above, radiation therapy, hormone antagonists, hormones and their releasing factors, thyroid and antithyroid drugs, estrogens and progestins, androgens, adrenocorticotropic hormone; adrenocortical steroids and their synthetic analogs; inhibitors of the synthesis and actions of adrenocortical hormones, insulin, oral hypoglycemic agents, and the pharmacology of the endocrine pancreas, agents affecting calcification and bone turnover: calcium, phosphate, parathyroid hormone, vitamin D, calcitonin, vitamins such as water-soluble vitamins, vitamin B complex, ascorbic acid, fat-soluble vitamins, vitamins A, K, and E, growth factors, cytokines, chemokines, muscarinic receptor agonists and antagonists; anticholinesterase agents; agents acting at the neuromuscular junction and / or autonomic ganglia; catecholamines, sympathomimetic drugs, and adrenergic receptor agonists or antagonists; and 5- hydroxytryptamine (5-HT, serotonin) receptor agonists and antagonists. Other suitable therapeutic agents for coadministration with immunotherapeutics with enhanced efficacy described herein also include agents for pain and inflammation such as histamine and histamine antagonists, bradykinin and bradykinin antagonists, 5- hydroxytryptamine (serotonin), lipid substances that are generated by biotransformation of the products of the selective hydrolysis of membrane phospholipids, eicosanoids, prostaglandins, thromboxanes, leukotrienes, aspirin, nonsteroidal anti-inflammatory agents, analgesic-antipyretic agents, agents that inhibit the synthesis of prostaglandins and thromboxanes, selective inhibitors of the inducible cyclooxygenase, selective inhibitors of the inducible cyclooxygenase-2, autacoids, paracrine hormones, somatostatin, gastrin, cytokines that mediate interactions Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 involved in humoral and cellular immune responses, lipid-derived autacoids, eicosanoids, β- adrenergic agonists, ipratropium, glucocorticoids, methylxanthines, sodium channel blockers, opioid receptor agonists, calcium channel blockers, membrane stabilizers and leukotriene inhibitors. Additional therapeutic agents contemplated for co-administration with immunotherapeutics with enhanced efficacy described herein include diuretics, vasopressin, agents affecting the renal conservation of water, rennin, angiotensin, agents useful in the treatment of myocardial ischemia, anti-hypertensive agents, angiotensin converting enzyme inhibitors, β-adrenergic receptor antagonists, agents for the treatment of hypercholesterolemia, and agents for the treatment of dyslipidemia. Other therapeutic agents contemplated for co-administration with immunotherapeutics with enhanced efficacy described herein include drugs used for control of gastric acidity, agents for the treatment of peptic ulcers, agents for the treatment of gastroesophageal reflux disease, prokinetic agents, antiemetics, agents used in irritable bowel syndrome, agents used for diarrhea, agents used for constipation, agents used for inflammatory bowel disease, agents used for biliary disease, agents used for pancreatic disease. Therapeutic agents used to treat protozoan infections, drugs used to treat Malaria, Amebiasis, Giardiasis, Trichomoniasis, Trypanosomiasis, and / or Leishmaniasis, and / or drugs used in the chemotherapy of helminthiasis. Other therapeutic agents include antimicrobial agents, sulfonamides, trimethoprim-sulfamethoxazole quinolones, and agents for urinary tract infections, penicillins, cephalosporins, and other, β-lactam antibiotics, an agent comprising an aminoglycoside, protein synthesis inhibitors, drugs used in the chemotherapy of tuberculosis, mycobacterium avium complex disease, and leprosy, antifungal agents, antiviral agents including nonretroviral agents and antiretroviral agents. Examples of therapeutic antibodies that can be combined with immunotherapeutics with enhanced efficacy described herein include but are not limited to anti-receptor tyrosine kinase antibodies (cetuximab, panitumumab, trastuzumab), anti CD20 antibodies (rituximab, tositumomab), and other antibodies such as alemtuzumab, bevacizumab, and gemtuzumab. Moreover, therapeutic agents used for immunomodulation, such as immunomodulators, immunosuppressive agents, tolerogens, and immunostimulants are contemplated by the methods herein. In addition, therapeutic agents acting on the blood and the blood-forming organs, Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 hematopoietic agents, growth factors, minerals, and vitamins, anticoagulant, thrombolytic, and antiplatelet drugs. EXPERIMENTAL Methods Cell Lines and General Cell Culture A20 murine B cell lymphoma tumor (ATCC) was used. Different human B cell lymphoma and leukemia were used including diffuse large B-cell lymphoma (DLBCL): OCI- Ly8.Luci.Chili, and OCI-Ly8.CD19KO.Luci.Chili; leukemia (NALM6.Luci.Chili). These human tumor cell lines were obtained originally from Dr. Jun Huang’s lab (University of Chicago). Cells were cultured at a concentration of 1 × 106cells / mL of standard culture media (RPMI 1640 (Gibco) + 10% fetal bovine serum albumin (FBS), 1% penicillin / streptomycin, 1% HEPES, 1% GlutaMAX, 1% Sodium pyruvate, 1% non-essential amino acids and 0.1µM 2-mercaptoethanol) at 37°C in 5%CO2 incubator. All cell lines were tested for Mycoplasma contamination (Lonza). Virus Production and Transduction of CAR-Engineered Murine and Human T Cells Replication-defective, second-generation lentiviral vectors were produced using Lenti-X 293T cells (Takara Bio). Approximately 5 × 106to 7 × 106cells were plated in T75 in standard culture media (DMEM + 10%FBS, without antibiotics) and incubated for 48 hours at 37°C. Then, cells were transfected with hCD19CAR lentiviral vector using a combination of Polyethylenimine (PEI) (Polysciences #23966-2) (64 μL, Invitrogen); pMD2.G (2 μg), and pCMV-dR8.2-dvpr (6 μg) packaging plasmids; and 8 μg of expression plasmid (CAR). PEI and plasmid DNA were diluted in 0.8 mL Opti-MEM (Thermo Scientific # 31985062) media prior to transfer into lentiviral production flasks. At both 24- and 48-hours following transfection, culture media containing viral particles were harvested and concentrated using Lenti-X Concentrator (Takara Bio) following the manufacturer’s instructions. PBMC samples were either collected from healthy donor volunteers under an approved IRB #14-0221-CR009 or from patient apheresis samples provided by UChicago cellular therapy biobank under the approved IRB #18-0025. Human T cells were isolated from PBMC using the Pan T Cell Isolation Kit following the manufacturer’s instructions (Miltenyi Biotec). Purified T cells were activated by T-Activator CD3 / CD28 Dynabeads (ThermoFisher) at a ratio 1:1 beads / cell in the presence of human IL-2 (200-500 U / mL) and incubated at 37°C overnight. Activated T cells were spin-infected with Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 hCD19CAR lentiviral supernatant using protamine sulfate (Sigma-Aldrich # P3369-10G) at a multiplicity of infection (MOI) = 5. Human T cells were further expanded for 10-13 days in the presence of IL-2 before harvesting for in vitro and in vivo studies. Designing CAR Constructs Overexpressing BCL-2 HiFi Assembly (NEB) was used to combine CAR and BCL2 transgenes into the second- generation lentiviral transfer plasmid pHR. The P2A ribosomal skip peptide was inserted between CAR and BCL2 sequences for bicistronic expression under the same promoter. Assembled products were transformed into STABLE chemically-competent E. coli (NEB) and plated on carbenicillin antibiotic plates. Individual colonies were inoculated and plasmid DNA isolated (Qiagen). Size verification was carried out by restriction enzyme digestion and gel electrophoresis. Sanger sequencing spanning at minimum the transgene promoter through stop codon was used to select final colonies. Mutagenic primers were then used to introduce F104L, G101V, or F104C substitutions into the BCL2 gene via site-directed mutagenesis. Flow Cytometry Assays For surface marker staining, cells were resuspended in FACS staining buffer (PBS + 2% FBS), stained with fluorophore-conjugated surface antibodies for 20 minutes on room temperature, washed and re-suspended in FACS buffer before analysis. For intracellular staining, initially surface marker staining was performed, followed by fixation and permeabilization using the FoxP3 Transcription Factor Staining Buffer Set according to the manufacturer’s protocol (eBioscience). To monitor cell apoptosis, fluorescent caspase-3 / 7 substrates for detecting apoptosis in intact cells; NucView® Caspase-3 Enzyme Substrates (Biotium #10405) were used following the manufacturer's protocol. Data were acquired on an NovoCyte Penteon (Agilent). For bulk RNA sequencing, CAR+T cells were sorted using a BD FACSAria Fusion. All data analyses were performed using FlowJo 10.8.1 software (FlowJo, LLC) or NovExpress software (Agilent). Experimental Mice, Xenograft Mouse Model, In vivo Treatment and Imaging BALB / c mice on the CD45.1 background and immunodeficient nonobese diabetic (NOD)–severe combined immunodeficient (Scid) IL-2RƔ-null (NSG) mice were purchased from Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 the Jackson Laboratory. Splenocytes from CD45.1 mice (6-10 weeks old) treated with vehicle or venetoclax 5 days per weeks for 3 weeks (total 15 doses) were harvested and used to prepare murine CARTs for in vitro studies. Venetoclax was reconstituted in 10% ethanol, 30% PEG 400, and 60% Phosal and was administered via oral gavage at a dose of 50 mg / kg. To establish the OCI-Ly8 subcutaneous (SQ) xenograft mouse model, 5 × 106cells of OCI-Ly8 tumors were prepared in 100 μL of PBS containing 50% Matrigel (Corning) and implanted into the right flank of NSG mice (5-8 weeks old) via subcutaneous injection. Suboptimal doses of CARTs (3.5 × 106CAR+cells) were administered intravenously when tumor size reached ∼ 30-50 mm2. Tumors were measured three times week by caliper, and tumor size was calculated according to the equation: tumor volume = (L × W), where L is the longest axis of the tumor and W is the axis perpendicular to L. For SQ tumor model, experiment humane end point was achieved when tumor size reached ~ 350mm2. For systemic intravenous (i.v.) xenograft mouse model, 1 × 106cells of NALM6.Luci.Chilli tumors in 200 μL of PBS were injected in tail NSG mice vein. After there days mice were either untreated or treated with the minimum effective dose of vehicle of venetoclax pre-treated CARTs (1.5 × 106CAR+cells). NALM6 engraftment and progression was monitored by imaging using Spectrum IVIS bioluminescence imager and quantified using Living Image software v.4.7.3 (Perkin Elmer), or by a Lago X imager and quantified using Aura software v.4.0.7 (Spectral Instruments Imaging), after mice after being injected intraperitoneal with 150mg / kg D-Luciferin (PerkinElmer # 122799). Imaging was performed under isoflurane anesthesia. For this systemic tumor model, animals were monitored for signs of disease progression and humane endpoints such as xenoGVHD, as evidenced by >15% loss in body weight, fur loss, diarrhea, and disease-related hind limb paralysis. All mice were housed under specific pathogen-free conditions. All animal care and use were followed by NIH guidelines, and all experimental protocols were approved by the Institutional Animal Care and Use Committee of the University of Chicago. Cytokine Secretion Assays Killing assay was setup by co-culturing 0.1 × 106CAR T cells were co-cultured with 0.5 ×106tumor cells in 200 μl of complete T cell medium without IL-2 in a 96-well plate, all in triplicate. Forty-eight hours after co-culture, culture supernatants were collected, diluted 10- to 100-fold and analyzed for IFNγ, TNFα, IL-2 and Granzyme B using ELISA LEGEND MAX kits Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 (BioLegend). Absorbance readings were collected on SpectraMax iD5 Multi-Mode Microplate Reader (Molecular Devices). Seahorse Assays Seahorse Bioscience Analyzer XFe96 (Agilent) was used to perform metabolic analyses. 0.2 × 106cells were resuspended in Seahorse XF DMEM Medium supplemented with 10 mM glucose, 1 mM sodium pyruvate, 2 mM glutamine and plated on a Cell-Tak (Corning #354240)- coated microplate allowing the adhesion of CAR T cells. Mitochondrial activities were measured by the oxygen consumption rate (OCR) (pmol / min) using Seahorse Mito Stress Test kit (Agilent # 103015-100) with the real-time injections of oligomycin (2 μM), carbonyl cyanide 4- (trifluoromethoxy) phenylhydrazone (FCCP; 1 μM) and rotenone and antimycin (Rot / AA 0.5 μM). Glycolytic parameters were evaluated by measuring extracellular acidification rate (ECAR) (mpH / min) using Glycolysis Stress Test Kit (Agilent #103020-100) with the addition of glucose (10 mM), oligomycin (2 μM) and 2-deoxy-glucose (2-DG; 50 mM). Respiratory parameters were acquired and calculated according to the manufacturer’s instructions (Seahorse Bioscience). scRNA-seq and Analysis Single-cell suspensions of vehicle or venetoclax-pretreated CARTs of three healthy donors were prepared and stained with Zombie NIR Dye to sort live cells. Sorted cells were counted (Logos Biosystems, LUNA-FL), and 10000 live cells were partitioned into droplets for single-cell omics assays via Chromium Next GEM Single-Cell 5’Kit v2 (10x Genomics, 1000263). RNA-seq libraries were prepared according to manufacturer protocols. All libraries were quantified via the Qubit dsDNA HS Assay Kit (Invitrogen, Q32851), quality-checked for fragment sizes via High Sensitivity D5000 ScreenTapes (Agilent, 5067-5592), pooled, and sequenced (Illumina, NovaSeq 6000). The raw scRNA-seq data were preprocessed using the Cell Ranger software (version 7.1.0; 10X Genomics). Feature-barcode matrices were obtained after aligning reads to a custom GRCh38 human reference genome with the CAR sequence. Sample counts matrices were imported into R version 4.3.0 for further analysis in Seurat version 5.1.0. Cells with more than 15% mtRNA, less than 500 identified features, and more than 50,000 RNA reads were filtered out. Cell cycle state was computationally identified and regressed using Seurat’s CellCycleScoring and ScaleData functions. Doublets were computationally identified Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 and removed using the DoubletFinder algorithm. Filtered matrices were normalized and centered, and principal component analysis was applied. The harmony algorithm was used to remove donor and treatment effects in dimensional reduction. Uniform manifold approximation and projection (UMAP) dimensional reduction was applied on the first 20 principal components. Graph-based clustering was performed on the reduced data to generate distinct Seurat objects for CD4+ and CD8+ CAR T-cells. Differential expression analysis was performed using the Seurat FindMarkers function with Wilcoxon test and Bonferroni p-value correction. Factor analysis was performed with SPECTRA. Pseudobulk gene set enrichment analysis was performed by first applying Seurat’s FetchData function to procure counts matrices, then ranking genes according to the Wald statistic in DESeq2, and running the cluterProfiler implementation of GSEA against the MSigDB’s HALLMARK database. Bulk RNA-Sequencing and Analysis Single-cell suspensions of vehicle or venetoclax-pretreated CARTs of three healthy donors were prepared and stained with Zombie NIR Dye, CD4, CD8, CAR+GFP to sort CD4+CAR+, CD8+CAR+live T cells. RNA was extracted from ~ 0.8X106cells using RNeasy Micro Kit (Qiagen). RNA was quantified via the Qubit RNA HS Assay Kit (Invitrogen, Q32852), and quality-assessed for integrity via High Sensitivity RNA ScreenTape (Agilent, 5067-5579). 500 ng high-quality RNA with a RIN score > 7 of each sample was used for library preparation using NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs, E7760S). Libraries were prepared according to manufacturer protocols. All libraries were quantified via the Qubit dsDNA HS Assay Kit (Invitrogen, Q32851), quality-checked for fragment sizes via High Sensitivity D5000 ScreenTapes (Agilent, 5067-5592), pooled, and sequenced (Illumina, NovaSeq X). RNA-sequencing reads were processed with the nf-core / rnaseq pipline, with default settings on the University of Chicago CRI’s Randi HPC. All analysis was performed in R version 4.3.0. Gene counts were imported to R / Bioconductor package DESeq2 for differential expression analysis. Differential expression analysis was performed using DESeq2 and differentially expressed genes were identified by Benjamini-Hochberg adjusted p-values less than or equal to 0.05 and log2FoldChange greater than 0.58 (corresponding to a 1.5 fold change in expression). Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 Functional enrichment analysis was performed using R / Bioconductor package clusterProfiler with functions for gene set enrichment analysis (GSEA, fGSEA implementation) and overrepresentation analysis (enricher) in reference to MSigDB’s HALLMARK database and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Heatmap visualizations of Z-score scaled expression are based on TPM normalization of counts. Statistical Analysis Data analysis was performed using GraphPad Prism V.10. All in vitro data presented are representative of at least two independent experiments except for scRNA-seq (performed once with three biological replicates). Each human CART data set show at least 2-3 different donors. All comparisons between two groups were performed using a Student’s t-test. Comparisons among multiple groups were evaluated with one-way ANOVA followed by Tukey’s multiple testing correction. All results are represented as mean ± SD unless otherwise noted. Survival data were analyzed using the log-rank (Mantel–Cox) test. Data analysis was performed using GraphPad Prism v10. P values > 0.05 were considered statistically significant. Results Venetoclax directly enhances the effector function of murine CD19CARTs Reconstitution of naive T cells in mice treated continuously with venetoclax, following T cell depleted bone marrow transplantation, results in acquisition of T cell active like-naïve genotype, increasing anti-apoptotic protein expression and cell death resistance. Additionally, BCL-2 blockade by venetoclax mediated Treg plasticity toward a Th17-like Treg phenotype, resulting in increased IL17A and IFN-γ production that synergizes with anti-PD-1 checkpoint blockade to enhance the overall antitumor control in mice bearing the colorectal cancer MC38. Experiments were conducted during development of embodiments herein to determine the direct effect of venetoclax on T cell effector function using a validated antitumor adoptive T cell model, murine T cells engineered to express CD19 chimeric antigen receptor (mCD19CART) against CD19+tumor cells. In this context, CD45.1 mice either orally received vehicle or venetoclax 50mg / kg for 15 days (5 days / week for 3 weeks). Then, total T cells were isolated from mouse splenocytes, activated and transduced to express CD19CAR in the presence of vehicle or venetoclax ex vivo (Fig. 17). To study the effect of venetoclax on mCD19CART Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 cytotoxic function, a killing assay was used in which CARTs treated under different conditions were co-cultured with CD19+murine B cell lymphoma (A20) in different effector to tumor (E:T) ratios. Administration of venetoclax either in vivo, ex vivo or both demonstrates enhanced antitumor cytotoxicity compared to vehicle only condition (Fig. 17B). Notably, short-term treatment of murine T cells with venetoclax for 2 days during CART manufacturing is enough to exhibit superior tumor killing effect over vehicle-treated CARTs (Fig. 17C). By checking the CD19CAR expression and the memory profile of the CART product after the short-term treatment with vehicle or venetoclax, neither the CAR expression nor the T cell memory content was dramatically affected by treatment with venetoclax (Fig. 17D, E). These results indicate that venetoclax immunomodulates CARTs during manufacturing to possess enhanced effector-like characteristics. Venetoclax is a compatible BH3 mimetic with hCD19CAR T cell manufacturing Experiments were conducted during development of embodiments herein to determine whether the results in the murine CD19CART model could be established in a human CD19CART system. Given that BH3 mimetics were originally designed to induce cancer cell apoptosis, it was suspected that these compounds could initially interfere with the human CD19CART production, limit the cell proliferation and expansion and induce cell death. To test this, after activation and CD19CAR lentivirus transduction, healthy donor T cells were incubated with escalating doses of venetoclax during expansion from day 2 till harvesting on day 10-13 (Fig. 1A). Venetoclax shows minimal inhibitory effect on CD19CART proliferation measured by T cell count (Fig. 11B). Paradoxically, the very minimum doses of A-1331852, a BCL-XL Inhibitor, or ABT-263 (navitoclax), the first BH3 mimetics binding both BCL-2, BCL-XL results in a drastically low expansion of hCD19CART (Fig. 18A-B). Taken together, this data indicates that, under the ex-vivo treatment schedule, venetoclax is a suitable BH3 mimetic for combination with hCD19CART during manufacture. Venetoclax-pretreated hCD19CARTs show upregulated expression of anti-apoptotic BCL-2 family of proteins Treating murine T cells in vitro or in vivo in the setting of BCL-2 blockade by venetoclax results in increased expression of the anti-apoptotic BCL-2 family proteins BCL-2, BCL-XL, Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 MCL-1. To test if this phenomenon is consistent in human T cells, hCD19CARTs expanded in presence of vehicle or venetoclax were flow cytometrically analyzed for the expression of different anti-apoptotic BCL-2 family proteins. Aligned with the murine results, hCD19CARTs show upregulation of BCL-2, BCL-XL, MCL-1 in both CD4+and CD8+hCD19CARTs in a dose dependent manner with venetoclax dose at 800nM demonstrating the highest upregulation in the expression (Fig. 11C, D). Notably, A-133 and navitoclax-pretreated hCD19CARTs show increase in the BCL-2 expression but to less extent compared to the same venetoclax dose (Fig. 18C). Further assessment of the CART CD4 / CD8 content and memory phenotype indicates that venetoclax did not affect the CD4 / CD8 ratio, CAR expression or final memory population composition of the CART product compared to the vehicle-treated CARTs (Fig. 11E-G). Collectively, this indicates that pressuring the apoptotic machinery by continuously and specifically blocking BCL-2 results in a compensatory upregulation of different anti-apoptotic proteins without affecting CART phenotype and content. Venetoclax enhances hCD19CART cytotoxicity in vitro Treating conventional or double negative patient derived T cells with venetoclax enhanced their anti-leukemic activity. Additionally, the murine CART model shows better antitumor efficacy of venetoclax-pretreated mCD19CARTs. Experiments were conducted during development of embodiments herein to determine whether venetoclax-pretreated hCD19CARTs have better antitumor activity. To test this hypothesis, killing assays were performed using vehicle or venetoclax-pretreated hCD19CARTs cocultured with CD19+expressing human diffuse large B cell lymphoma (DLBCL) OCI-Ly8.Luci. Aligned with previous findings in mCD19CART, venetoclax-pretreated hCD19CARTs dose-dependently killed tumor cells more effectively than vehicle-pretreated hCD19CARTs. (Fig. 12A). Venetoclax was not present in culture with tumor cell targets ensuring that tumor cell death was not the result of direct BCL-2 drugging in tumor cells. The same effect was detected against CD19+acute lymphoblastic leukemia NALM6.Luci (Fig. 12B). To ensure that venetoclax induces antigen specific killing, the cytotoxicity assay was repeated using CD19 knock out OCI-Ly8 tumor cells (OCI- Ly8.CD19KO.Luci) (Fig. 19A). Venetoclax-pretreated hCD19CARTs fail to kill the CD19- cell line (Fig. 19B). Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 To test whether the enhanced cytotoxicity of venetoclax-pretreated hCD19CARTs is dependent on a certain co-stimulatory molecule, the killing assay was repeated against OCI- Ly8.Luci tumor cells using 4-1BB CD19CAR (used in the whole study) versus CD28 CD19CAR treated with vehicle or venetoclax. Additionally, to determine whether the effect is based on certain cytokines used during CART expansion, vehicle or venetoclax-pretreated CD19CARTs expanded was used in the presence of IL-2 (used in the whole study) versus CD19CARTs expanded in IL-7 / IL-15. Venetoclax-pretreated hCD19CARTs induced similar enhanced cytotoxicity in vitro regardless the type of the co-stimulatory molecule in the CD19CAR construct (Fig. 19C), or the cytokine used in the CAR T cell expansion (Fig. 19D). Pretreatment of hCD19CARTs with other BH3 mimetics such as A-133 is unable to enhance their killing efficacy (Fig. 18D). However, hCD19CARTs preconditioning with Navitoclax at low dose 25nM show enhanced antitumor effect, indicating that targeting BCL-2 is necessary for the observed improved potency of pretreated CARTs (Fig. 18D). Together, this data indicates that venetoclax, under specified manufacture conditions, can amplify the killing capacity of hCD19CARTs in an antigen specific manner. Advanced T cell effector function mediates venetoclax-pretreated hCD19CART enhanced cytotoxicity. To better understand the cytotoxic mechanisms that venetoclax induced to allow better antitumor activity of hCD19CARTs, experiments were conducted during development of embodiments herein to study the effector, persistence and exhaustion characteristics of venetoclax-pretreated hCD19CARTs upon specific tumor antigen stimulation. After 48 hours of tumor stimulation, increased effector activity of venetoclax-pretreated hCD19CARTs was detected, as reflected in dose-dependent increases in the cytokine release of IFN-γ, IL-2, TNF-α, and granzyme B (GZMB) (Fig. 12C). Without being bound by theory, it was contemplated that the induced effector function of venetoclax-pretreated hCD19CARTs is due to increased expansion of effector T cells. The memory profiling of CARTs after tumor stimulation indicates increase in central memory (TCM) in venetoclax-pretreated CD8+CARTs in the expenses of effector T cells (TEM) that is more enriched in vehicle-pretreated CARTs while no significant changes was detected in CD4+CARTs memory phenotype under the two treatment conditions (Fig. 12D). Venetoclax-pretreated hCD19CARTs show cell death resistance upon tumor Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 stimulation detected by higher percent of live CARTs (G2: 9.16% vs 2.31%), less dead CARTs (G1-CD8+: 8.81% vs 23.86%) compared to significant CD8+dead cells in vehicle-pretreated hCD19CARTs, and lower caspase3 / 7 enzymatic activity (Fig. 12E). By focusing on CD8+CARTs, it was found that venetoclax upregulates the expression of intracellular IFN-γ and surface death receptor ligand, TRAIL that known to be essential for CAR T cell cytotoxicity (Fig. 12F). Additionally, venetoclax-pretreated hCD19CARTs demonstrate lower expression of exhaustion markers LAG3, PD-1, TIM3, and TIGIT upon acute tumor stimulation compared to vehicle-pretreated hCD19CARTs (Fig. 12G). These results indicate that expansion of hCD19CARTs in the presence of venetoclax improves their effector function, exhaustion and cell death resistance upon activation by specific tumor antigen stimulation. BCL-2 mutation F104L or F104C located within BCL-2’s BH3-binding groove, rendered human leukemia and lymphoma cell lines resistant to venetoclax, but not yet observed in patient, while the BCL-2 mutant G101V was reported in chronic lymphocytic leukemia (CLL) patients. To evaluate whether the observed effect of venetoclax on CARTs is due to on-target effect on BCL-2, CD19CART vectors that either overexpress wild type BCL-2 (BCL-2 WT), or one of the BCL-2 mutants were designed to induce at least partial resistance to venetoclax. Profiling the effect of venetoclax was conducted on the CAR19_BCL-2 G101V and F104C mutants as they were reported to reduce the affinity to the drug ~ 18-50-fold more than the BCL-2 F104L that has 10-fold less binding affinity to venetoclax than the wild type BCL-2. Comparing the proliferation and expansion of CARTs under each construct, no significant difference was detected in the final product cell count between vehicle versus venetoclax-pretreated CARTs (Fig. 13A). Overexpression was confirmed of BCL-2 protein in all BCL-2 expressing constructs and observed further upregulation of BCL-2 in response to venetoclax (Fig. 13B). The delta change of venetoclax-induced BCL-2 was higher in CAR19_BCL-2 WT compared to the mutant BCL-2. The expression of BCL-XL and MCL-1 reflects the reduced response to the effect of venetoclax in case of using CAR19_BCL-2 mutants compared to the CAR19_BCL-2WT, with much reduced response observed in the clinically reported BCL-2 mutant, G101V (Fig. 13C, D). To test whether the killing capacity of CAR19_BCL-2 mutants are affected by expanding cells in presence of venetoclax, it was assured that each construct upon treatment of vehicle or venetoclax has same CAR+level (Fig. 13E). Then, a killing assay was setup to compare the effect of venetoclax in the effector function of wildtype vs mutant BCL-2 overexpressing Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 CARTs. It was found that the venetoclax- improved killing efficacy was partially diminished in the CAR19_BCL-2 mutants compared to CAR19_BCL-2WT with CAR19_BCL-2 G101V showing the greatest significant loss of venetoclax-induced CART cytotoxicity (Fig. 13F). Furthermore, profiling the CARTs for exhaustion upon tumor stimulation indicates that CARTs expressing the BCL-2 mutants have lost the venetoclax-exhaustion control feature in a manner correlated to the decrease of the mutant binding affinity to venetoclax (Fig. 13G). Collectively, these results confirm that the enhanced effect of venetoclax on CARTs is leaded by direct and on-target effect on BCL-2 protein that significantly affected by the binding affinity of the drug to the protein. Moreover, venetoclax effect on CARTs is beyond the known apoptotic features of targeting BCL-2 by venetoclax and involving more T cell signaling pathways. Venetoclax induces universal transcriptional alterations of hCD19CARTs Since the enhanced cytotoxicity observed with venetoclax-pretreatead CARTs was neither accompanied by changes in the CART product content of CD4+ / CD8+nor a distinctive change in the memory phenotype of the product but associated with enhanced effector function while maintaining higher central memory phenotype, it was contemplated that venetoclax induces disparate gene expression reprograming that leads to acquire effector like characteristics that are functional upon tumor stimulation. To clarify whether venetoclax-induced the expansion of distinct T cell subpopulation or specific T cell states with distinctive transcriptional features, the expansion timeline was followed (Fig. 11A) and performed scRNA-seq on three CART products prepared from three healthy donors’ PBMC and harvested on day 13. Additionally, CD4+CAR+and CD8+CAR+were sorted from CART products prepared from other three healthy donors’ PBMC for bulk RNA sequencing to get transcription profiling of vehicle versus venetoclax-pretreated CART from a total of six healthy donors. Venetoclax led to distinctive transcriptional profiles as principal component analysis (PCA) show that ventoclax-pretreated CD4+and CD8+results in pronounced changes for each donor (Fig. 14A) with homogenous expression of differentially expressed genes (DEG) within each treatment group (Fig. 14E). Cell clusters within CD4+and CD8+CART product were computationally identified on a set of harmony dimensions, integrated to remove donor and treatment effect, clustered using the Louvain algorithm and projected to UMAP (Fig. 14B). Cells under vehicle and venetoclax Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 conditions segregates into the same number of clusters in both CD4+and CD8+CART (Fig. 14B). Despite some clusters under CD8+CART show different extent of expansion with more predominant proliferative and central memory clusters in venetoclax-pretreated CART compared to vehicle, this difference in cluster proportions is not significant among the tested three donors CART products (Fig. 14C). scRNA-seq identifies around 514 and 1078 DEGs in CD8+and CD4+T cells, respectively, with a log2FoldChange of >0.58 and a BH-adjusted p-value threshold of 0.05 (Fig. 14E). Among the different components and regulators of the apoptotic machinery that significantly changed its expression, BCL-2 has significantly higher expression across all venetoclax-pretreated CD4+and CD8+clusters relative to vehicle-pretreated clusters (Figs. 14D, 20A). Gene set enrichment analysis (GSEA) against MSigDB’s HALLMARK pathways demonstrates that venetoclax promotes enrichment of similar pathways between CD4+and CD8+CARTs suggesting a shared transcription program associated with T cell activation, effector function, cell cycle, survival and metabolism including positive enrichment for IL- 2 / STAT5, PI3K / AKT / MTOR, MTORC1, MYC targets, TNFα / NFKB signaling, fatty acid metabolism and oxidative phosphorylation (OXPHOS) and negative enrichment for glycolysis and hypoxia (Figs. 14F, 20B). Significant enrichment of these pathways is consistent across clusters in both CD4+and CD8+CART product (Fig. 14G). In line with factor analysis, gene set enrichment analysis (GSEA) has also demonstrated the upregulation of transcriptomic signature of IFN-α, IFN-γ and TNF-α signaling (Fig. 20C). Given common enriched pathways between pseudobulked scRNAseq and bulk RNA-seq, experiments were conducted during development of embodiments herein to identify whether select genes were core contributors to these enrichments. Genes were identified that were most frequently present in the leading edge of each enrichment across our bulk RNA-seq and scRNA-seq data in the 6 donors. Indeed, it was observed that many genes are shared across pathways and between sequencing datasets, indicating that a core transcriptomic mechanism exists across pathways under venetoclax treatment (Fig. 14H). Altogether these data indicate that the enhanced effect observed with venetoclax is due to broad-spectrum transcriptional reprogramming of whole T cells, not a specific subset and that this transcriptional reprogramming relates to improved T cell signaling, functionality, and metabolic fitness of the CART product. Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 The antitumor effect of venetoclax-pretreated hCD19CART is mediated by improved T cell signaling Recent data suggests that PI3K / AKT and JAK / STAT5 signaling are key determinants of CART fitness, as defined by increased proliferation, effector function, memory and survival. Homeostatically expanding naïve murine CD4+and CD8+T cells following during venetoclax administration leads to acquiring active like effector function marked by upregulation of PI3K / AKT, Jak-STAT and downregulation of MAPK and FOXO signaling pathways. Aligned with our published murine data, further GSEA of scRNAseq reveals a significant upregulation of PI3K / AKT / MTOR and IL-2 / STAT5 signaling (Fig. 15A, C). To confirm these results, CARTs pretreated with vehicle or venetoclax were intracellularly stained to detect the active phosphorylated form of key regulator of each pathway. pAKT and pSTAT5 were upregulated in both CD4+and CD8+CARTs after expansion in presence of venetoclax (Fig. 15B, D). Pharmacologic inhibitors were used to further evaluate how these pathways are involved in the improved functionality of venetoclax-pretreated CARTs. Following the timeline indicated (Fig. 11A), incubating venetoclax-pretreated CARTs harvested on day 13 with titrated doses of STAT5-inhibitor AC-4-13 (JAK / STAT5i) for 6 or 24 hours then setting up killing assay led to diminished cytotoxic efficacy of venetoclax-pretreated CARTs in a time and dose dependent manner (Figs. 15E, 21A). Similarly, incubating CARTs overnight with AKT Inhibitor VIII (AKTi) for 24 hours results in decline in the antitumor efficacy of venetoclax-pretreated CARTs in a dose dependent manner (Fig. 11F). Venetoclax promotes hCD19CARTs metabolic fitness Besides being reported for increasing reactive oxygen species (ROS) in cancer cells, short-term treatment with venetoclax was reported to enhance the antileukemic activity of T cells by inducing ROS through impairing respiratory chain supercomplex formation to increase T cell effector function without affecting other cell metabolic parameters. HALLMARK pathway analysis illustrates positive enrichment of OXPHOS and ROS pathway and negatively enriched glycolysis in both CD4+and CD8+CARTs (Fig. 14F). GSEA validates these data and shows upregulation of genes regulated by PPARGC1A as well (Fig. 15G). PPARGC1A gene encodes for PGC1α, a key metabolic regulator that promotes mitochondrial biogenesis, OXPHOS, fatty acid β-oxidation, thus enhancing T cell fitness, memory formation and antitumor potency. Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 Therefore, experiments were conducted during development of embodiments herein to test whether there is interconnection between these transcription alterations induced by venetocalx and the actual metabolic profile of CART product. Using Mito stress assay, the oxygen consumption rate (OCR) of vehicle and venetoclax-pretreated CARTs was examined at the end of production on day 13. Initially, basal OCR was measured, followed by addition of an inhibitor of ATP synthesis (oligomycin), an uncoupling ionophore (FCCP) and complex I / III blockers (rotenone with antimycin A) respectively to decipher the relative contribution of mitochondrial and non-mitochondrial mechanisms of oxygen consumption. Venetoclax significantly increases basal OCR and maximal respiration and spare respiratory capacity (SRC) (Fig. 15I). Additionally, the extracellular acidification rate (ECAR) was measured to determine the glycolysis status and lactic acid level in cells. While glycolysis is not affected, venetoclax- pretreated CARTs show elevated glycolytic capacity and glycolytic reserve. Several studies illustrate that TCM cells display higher basal OCR and SRC compared to TEM and TEMRA through increased dependence on fatty acid oxidation (FAO). Further, several metabolic strategies that inhibit glycolysis, promote OXPHOS or induce PGC1α have been reported to maintain less differentiated T cells and enhance the efficacy of CARTs. Therefore, the observed venetoclax- induced metabolic features indicate enhanced metabolic fitness of the CART product that promotes memory over effector metabolic status while having capability of meeting the metabolic requirements of effector function under stress or upon tumor stimulation. Additionally, incubating venetoclax-pretreated CARTs with escalating doses of the antioxidant N-Acetyl-L-cysteine (LNAC) for 6 or 24 hours before co-culturing with tumor shows no effect on the venetoclax-induced killing capacity of CARTs. This result demonstrates that ROS induction is not involved in the observed effect of ventoclax on CARTs in the system (Fig. 21B). Venetoclax-pretreated hCD19CARTs exhibits boosted therapeutic activity against leukemia and lymphoma in vivo Given the observed improvement of venetoclax-pretreated CARTs in tumor cell lysis in vitro, experiments were conducted during development of embodiments herein to investigate whether this can be reflected into augmented antitumor efficacy in vivo in the absence of venetoclax direct administration. In this context, NALM6 tumors were intravenously injected in Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 NOD-scid IL2Rgnull(NSG) mice. On day 3, mice were randomly divided in to three groups that either untreated or treated with vehicle or venetoclax-pretreated CARTs (Fig. 16A). Mice received venetoclax-pretreated CARTs show delayed tumor growth as observed around day 9 to day 16 after CART infusion as detected by in vivo bioluminescent imaging (BLI) (Fig. 16B). Mice subjected to control CARTs show modest tumor control over the untreated, while mice received venetoclax-pretreated CARTs show more sustained delay in tumor growth over time and enhanced overall survival over the other treatment conditions (Fig. 16C). To figure out whether these findings apply to other tumor models, mice with established subcutaneous OCI.Ly8 tumors were either untreated, received vehicle or venetoclax-pretreated CARTs. Tumors were measured to monitor tumor growth and survival. While mice treated with vehicle-pretreated CARTs show slight tumor control over the untreated group, administration of venetoclax-pretreated CARTs demonstrate significantly improved antitumor control and prolonged survival (Fig. 16D, E). Venetoclax augments the efficacy of hCD19CARTs prepared from patient apheresis bulb in vivo. Experiments were conducted during development of embodiments herein to determine whether venetoclax enhances the efficacy of CARTs prepared from cancer patient-derived T cells. PBMCs from six patients at the time of commercial CAR T cell apheresis were provided by University of Chicago Cellular Therapy Biobank to test the effect of venetoclax in lab-grade CART. Samples were divided into two groups based on 1- and 3-month follow up on responding to the commercial CART product the patient received; Three samples were reported as partial responding (PR) while the other three samples were from complete responders (CR) (Table 1). The samples against healthy donor (HD) PBMC were first assayed to determine the CD4:CD8, regulatory T cells (Tregs) content, T cell memory phenotype, exhaustion and anti-apoptotic machinery status. Most of CR-samples contain the highest percent of CD4 and consequently the lowest percent of CD8 while most of PR-samples show higher percent of CD8 T cells (Fig. 22A). T cell memory profiling reveals that HD-samples tend to possess T cells in naïve (TN), to central memory (TCM) status, PR-samples tend to have cells in a more differentiated less naïve status while CR-samples memory status fall in the middle between HD and PR-samples which also reflected by the expression level of CD62L,CD127 and TCF1 HD > CR > PR (Fig. 22B, F, Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 G). Additionally, PR-samples contains % of Tregs comparable to HD but higher than the CR- samples (Fig. 22C). Most of exhaustion surface markers like LAG3 and TIM3 and the intracellular marker TOX indicate that PR-samples contain more exhausted cells than HD and CR-samples (Fig. 22D, E). Regarding the apoptotic status, CR-samples show the highest expression level of the anti-apoptotic proteins BCL-2, BCL-XL and MCL-1 (Fig. 22H) which is aligned with the recent finding that BCL-2 expression in patient apheresis is positively correlated to CART response. Table 1. 30-day 3-month Lines of se Following the manufacture timeline in Fig. 11A, CD19CAR was successfully we transduced to all samples. Transduction efficiency was comparable between HD and CR- samples, but PD-samples show lower transduction efficiency (Fig. 23A). While HD-samples show higher content of CD8+T cells and CR-samples has the lowest content of CD8+reflecting the starting material status, PR-samples show unexpected shrink of CD8+population in favor of more expanded CD4+T cells (Fig. 23B). Tregs content was comparable between CR- and PR- samples which is slightly higher than HD-samples (Fig. 23C). Venetoclax demonstrates ability to enhance the antitumor efficacy of CARTs in vitro and in vivo against OCI-Ly8 tumors regardless of the memory, exhaustion, anti-apoptotic status and CD4:CD8 content in the starting Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 material of CART product of each sample category (Fig. 16F-I). 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Claims

Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 CLAIMS 1. An immunotherapeutic composition comprising a lymphocyte (i) engineered to express a chimeric antigen receptor (CAR), and (ii) treated with a B-cell lymphoma 2 (BCL-2) inhibitor.

2. A method of enhancing a chimeric antigen receptor (CAR) lymphocyte comprising treating the CAR lymphocyte with a B-cell lymphoma 2 (BCL-2) inhibitor.

3. An immunotherapeutic composition comprising a chimeric antigen receptor (CAR) lymphocyte enhanced by the method of claim 2.

4. A method of preparing an immunotherapeutic composition comprising: (a) obtaining lymphocytes from a subject; (b) activating the lymphocytes; (c) inducing the lymphocytes to express a chimeric antigen receptor (CAR) to produce CAR lymphocytes; and (d) treating the CAR lymphocytes with a B-cell lymphoma 2 (BCL-2) inhibitor.

5. An immunotherapeutic composition prepared by the method of claim 4.

6. The immunotherapeutic composition of one of claims 1, 3, and 5 or method of claim 2 or 4, wherein the CAR comprises an extracellular antigen recognition domain capable of binding to a target selected from the group consisting of TSHR, CD19, CD123, CD22, BAFF-R, CD30, CD171, CS-1, CLL-1, CD33, EGFRvIII, GD2, GD3, BCMA, GPRC5D, Tn Ag, PSMA, ROR1, FLT3, FAP, TAG72, CD38, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, mesothelin, IL-1 1Ra, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-0, SSEA-4, CD20, AFP, Folate receptor α, ERBB2 (Her2 / neu), MUC1, EGFR, CS1, CD138, NCAM, Claudin18.2, Prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gp100, bcr-ab1, tyrosinase, EphA2, Fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, Folate receptor R, TEM1 / CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD 179a, ALK, polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP,Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 WT1, NY-ESO-1, LAGE-1a, MAGE-A1, legumain, HPV E6 / E7, MAGE A1, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, prostate specific protein, survivin and telomerase, PCTA-1 / Galectin 8, MelanA / MART1, Ras mutant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2ETS fusion gene), NA17, PAX3, androgen receptor, Cyclin B1, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES 1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal tract carboxylesterase, mut hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, PD1, PDL1, PDL2, TGFβ, APRIL, and NKG2D.

7. The immunotherapeutic composition or method of claim 6, wherein the extracellular antigen recognition domain capable of binding to CD19.

8. The immunotherapeutic composition or method of claim 6, wherein the extracellular antigen recognition domain is a short-chain variable fragment (scFv).

9. The immunotherapeutic composition or method of claim 6, wherein the CAR further comprises a transmembrane domain and intracellular signaling domain.

10. The immunotherapeutic composition or method of claim 6, wherein the CAR further comprises a costimulatory domain.

11. The immunotherapeutic composition of one of claims 1, 3, and 5-10 or method of claim 2 or 4, wherein the BCL-2 inhibitor is a BCL-2 homology 3 (BH3) mimetic.

12. The immunotherapeutic composition or method of claim 11, wherein the BH3 mimetic is selected from venetoclax, ABT-199 (4-[4-[[2-(4-Chlorophenyl)-4,4-dimethylcyclohex-1-en-1- yl]methyl]piperazin-1-yl]-N-[[3-nitro-4-[[(tetrahydro-2H-pyran-4- yl)methyl]amino]phenyl]sulfonyl]-2-[(1H-pyrrolo[2,3-b]pyridin-5-yl)oxy]benzamide), ABT-737 (4-[4-[[2-(4-chlorophenyl)phenyl]methyl]piperazin-1-yl]-N-[4-[[(2R)-4-(dimethylamino)-1- phenylsulfanylbutan-2-yl] amino]-3-nitrophenyl]sulfonylbenzamide) (navitoclax), ABT-263Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 ((R)-4-(4-((4′-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1′-biphenyl]-2-yl)methyl)piperazin-1- yl)-N-((4-((4-morpholino-1-(phenylthio)butan-2-yl)amino)- 3((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide), GX15-070 (obatoclax mesylate, (2Z)- 2-[(5Z)-5-[(3,5-dimethyl-1H-pyrrol-2-yl)methylidene]-4-methoxypyrrol-2-ylidene]indole; methanesulfonic acid))), 2-methoxy-antimycin A3, YC137 (4-(4,9-dioxo-4,9- dihydronaphtho[2,3-d]thiazol-2-ylamino)-phenyl ester), pogosin, ethyl 2-amino-6-bromo-4-(1- cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate, Nilotinib-d3, TW-37 (N-[4-[[2-(1,1- Dimethylethyl)phenyl]sulfonyl]phenyl]-2,3,4-trihydroxy-5-[[2-(1- methylethyl)phenyl]methyl]benzamide), Apogossypolone (ApoG2), HA14-1, AT101 (gossypol), sabutoclax, gambogic acid, and G3139 (Oblimersen).

13. The immunotherapeutic composition of one of claims 1, 3, and 5-10 or method of claim 2 or 4, wherein the BCL-2 inhibitor inhibits expression of BCL-2.

14. The immunotherapeutic composition of claim 13, wherein the BCL-2 inhibitor is an shRNA, a miRNA, a morpholino, a ribozyme, an antisense nucleic acid molecule, or a CRISPR- Cas9-based construct.

15. The immunotherapeutic composition of one of claims 1, 3, and 5-14 or method of claim 2 or 4, wherein the lymphocyte(s) are T cells.

16. The immunotherapeutic composition of one of claims 1, 3, and 5-14 or method of claim 2 or 4, wherein the lymphocyte(s) are NK cells.

17. A method of treating a subject suffering from cancer comprising administering to the subject the immunotherapeutic composition of one of claims 1, 3, 5, and 6-16.

18. The method of claim 17, wherein the subject suffers from a blood or bone marrow cancer.Attorney Docket No. UCHI-42274.601 Client Ref. No. 24-T-003 19. The method of claim 18, wherein the subject suffers from a leukemia, lymphoma or myeloma.

20. The method of claim 19, wherein the subject suffers from B-cell acute lymphoblastic leukemia (B-ALL), chronic lymphocytic leukemia (CLL), or B-cell non-Hodgkin lymphoma (B- NHL), 21. The method of claim 17, wherein the subject suffers from a solid tumor cancer.

22. The method of claim 17, further comprising co-administering one or more additional cancer treatments.

23. The method of claim 20, wherein the one or more additional cancer treatments are selected from chemotherapies, immunotherapeutics, radiation treatment, and surgery.

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