PSCA car-t cells

Abstract

To provide T cell compositions and methods for treating diseases and disorders associated with presence of tumor cells that express a prostate stem cell antigen.SOLUTION: In one aspect, a method for treating a human subject diagnosed so as to suffer from a disease associated with presence of one or more solid tumors expressing a prostate stem cell antigen (PSCA) comprises administering, to the subject, 0.3×106 cells / kg to about 9×106 cells / kg of modified T cells comprising (i) a first polynucleotide encoding an inducible MyD88 / CD40 polypeptide and (ii) a second polynucleotide encoding a prostate stem cell antigen chimeric antigen receptor (PSCA-CAR) polypeptide.SELECTED DRAWING: None

Description

[Technical field]

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 62 / 779,394, filed December 13, 2018, which is hereby incorporated by reference in its entirety. Sequence Listing

[0002] This application contains a Sequence Listing that has been submitted electronically in ASCII format, which is hereby incorporated by reference in its entirety. Said ASCII copy, created on December 4, 2019, is named 14562-068-228_SL.txt and is 61,765 bytes in size. Technical Field

[0003] The present technology relates generally to the field of immunology, and in part to T cell compositions and methods for treating diseases and disorders associated with the presence of tumor cells that express prostate stem cell antigen. [Background technology]

[0004] Prostate stem cell antigen (PSCA) is a glycosylphosphatidylinositol-anchored cell surface protein with 123 amino acids. PSCA is expressed poorly in normal epithelial cells of the prostate, bladder, kidney, skin, esophagus, stomach, eye and placenta, but PSCA is upregulated in cancers of the pancreas, prostate, stomach, ovary and bladder (Abate-Daga D 2014 Hum Gene Ther 25: 1003-1012). PSCA expression positively correlates with advanced clinical stage and metastasis of prostate cancer (Saeki N 2010 Cancer Res 16: 3533-3538).

[0005] PSCA represents a potential target for anti-cancer immunotherapy. The safety and clinical activity of monoclonal antibodies against PSCA (AGS-1C4D4 or AGS-PSCA) have been evaluated in phase I and II clinical trials for the treatment of advanced pancreatic cancer (Wolpin BM 2013 Ann Oncol 24: 1792) and prostate cancer (Antonarakis ES 2012 Cancer Chemother Pharmacol 69: 763-771, Morris MJ 2012 Ann Oncol 23: 2714-2719). In approximately 200 subjects, treatment with anti-PSCA antibodies showed low toxicity and moderate efficacy at doses of 48 mg / kg or less. This favorable safety profile has been attributed to the limited expression of PSCA on normal tissues (Abate-Daga D 2014 Hum Gene Ther 25: 1003-1012), which may help to reduce on-target, off-tumor side effects. Collectively, these data provide clinical evidence that PSCA is an interesting and valid target for therapeutic intervention.

[0006] Inducible PSCA CAR-T cells may be a useful type of immunotherapy for the treatment of PSCA-expressing solid tumor malignancies. Clinical data show enhanced T cell proliferation, persistence and in vivo anti-cancer activity compared to conventional CAR-T therapies (Foster AE 2017 Mol Ther 25: 2176-2188, Mata M 2017 Cancer Discov 7: 1306-1319). So far, meaningful clinical activity of conventional CAR-T cell immunotherapy has been limited to hematological malignancies, while efficacy in solid tumor indications may be limited by generally poor CAR-T cell persistence and thus lack of sustained anti-tumor activity over time. Adding an inducible "on" switch to anti-PSCA CAR-T cells controls activation and proliferation signals, thereby improving the persistence of modified T cells as well as prolonging potential anti-tumor efficacy.

[0007] Certain embodiments are further described in the following description, examples, claims and figures. [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] Abate-Daga D 2014 Hum Gene Ther 25: 1003-1012 [Non-Patent Document 2] Saeki N 2010 Cancer Res 16: 3533-3538 [Non-Patent Document 3] Wolpin BM 2013 Ann Oncol 24: 1792 Summary of the Invention [Means for solving the problem]

[0009] In one aspect, a method is provided for treating a human subject diagnosed with a disease associated with the presence of one or more solid tumors expressing prostate stem cell antigen (PSCA), comprising administering to the subject 0.3×10 mAbs containing (i) a first polynucleotide encoding an inducible MyD88 / CD40 polypeptide and (ii) a second polynucleotide encoding a prostate stem cell antigen-chimeric antigen receptor (PSCA-CAR) polypeptide. 6 cells / kg ~ approx. 9×10 6 Provided herein are methods comprising administering cells / kg of the modified T cells.

[0010] In some embodiments, the methods for treating a human subject provided herein include administering 0.1×10 unmodified polyclonal T cells to a subject. 6 cells / kg~30×10 6The method further comprises administering an amount between about 20% and about 90% of the T cells are modified T cells. In certain embodiments, the subject is infused with a heterogeneous T cell population comprising modified and unmodified T cells, and about 20% to about 80% of the T cells are modified T cells. In some embodiments, the subject is infused with a heterogeneous T cell population comprising modified and unmodified T cells, and about 20% to about 70% of the T cells are modified T cells. In some embodiments, the subject is infused with a heterogeneous T cell population comprising modified and unmodified T cells, and about 20% to about 60% of the T cells are modified T cells. In some embodiments, the subject is infused with a heterogeneous T cell population comprising modified and unmodified T cells, and about 20% to about 50% of the T cells are modified T cells.

[0011] In certain embodiments, the T cells are about 20×10 per milliliter. 6 ~120×10 6 The cells are administered to the subject in a suitable dilution for injection into the subject, comprising:

[0012] In some embodiments, the methods for treating a human subject provided herein further comprise administering multiple doses of rimiducid to the subject. In certain embodiments, rimiducid is administered to the subject once daily. In other embodiments, rimiducid is administered to the subject three times per week. In still other embodiments, rimiducid is administered to the subject twice per week. In some embodiments, rimiducid is administered to the subject once per week. In some embodiments, rimiducid is administered to the subject once every other week. In other embodiments, rimiducid is administered to the subject twice per month.

[0013] In certain embodiments, each administration of rimducside to a subject is about 0.4 mg / kg of the subject's body weight. In certain embodiments, rimducside is administered to a subject once a week, and each administration of rimducside to a subject is about 0.4 mg / kg of the subject's body weight. In other embodiments, rimducside is administered to a subject once a day, and each administration of rimducside to a subject is about 0.4 mg / kg of the subject's body weight. In yet other embodiments, rimducside is administered to a subject three times a week, and each administration of rimducside to a subject is about 0.4 mg / kg of the subject's body weight. In some embodiments, rimducside is administered to a subject twice a week, and each administration of rimducside to a subject is about 0.4 mg / kg of the subject's body weight. In some embodiments, rimducside is administered to a subject once every other week, and each administration of rimducside to a subject is about 0.4 mg / kg of the subject's body weight.

[0014] In some embodiments of any one of the methods for treating a human subject provided herein, the solid tumor is pancreatic cancer, gastric cancer, or prostate cancer.

[0015] In some embodiments of any one of the methods for treating a human subject provided herein, the inducible MyD88 / CD40 polypeptide comprises: i) two multimeric ligand-binding domains; ii) a MyD88 polypeptide, or a truncated MyD88 polypeptide lacking the TIR domain, and iii) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain In certain embodiments, each multimeric ligand binding region comprises an FKBP12 variant polypeptide. In some embodiments, each FKBP12 variant polypeptide comprises an amino acid substitution at position 36 and binds to a multimeric ligand with higher affinity than a wild-type FKBP12 polypeptide. In certain embodiments, the amino acid substitution at position 36 is selected from the group consisting of valine, isoleucine, leucine and alanine. In some embodiments, each multimeric ligand binding region is an FKB12v36 region.

[0016] In some embodiments, the truncated MyD88 polypeptide has the amino acid sequence of SEQ ID NO: 21, or a functional fragment thereof.

[0017] In certain embodiments, the CD40 cytoplasmic polypeptide comprises the amino acid sequence of SEQ ID NO:23, or a functional fragment thereof.

[0018] In some embodiments of any one of the methods provided herein, the PSCA-CAR polypeptide comprises an antigen recognition portion, a transmembrane region, and a T cell activation molecule. In certain embodiments, the antigen recognition portion is derived from a PSCA antibody selected from the group consisting of 1G8, 2A2, 2H9, 3C5, 3E6, 3G3, 4A10, and A11, where 1G8, 2A2, 2H9, 3C5, 3E6, 3G3, and 4A10 are produced by hybridomas designated HB-12612, HB-12613, HB-12614, HB-12616, HB-12618, HB-12615, and HB-12617, respectively, deposited at the American Type Culture Collection.

[0019] In some embodiments, the antigen recognition portion comprises the complementarity determining regions (CDRs) of the heavy chain variable domain and the light chain variable domain of an amino acid sequence selected from the group consisting of SEQ ID NOs: 65, 67, 69 and 71.

[0020] In other embodiments, the antigen recognition portion comprises a heavy chain variable amino acid sequence and a light chain variable amino acid sequence selected from the group consisting of SEQ ID NOs: 51 and 49, SEQ ID NOs: 55 and 53, SEQ ID NOs: 57 and 59, and SEQ ID NOs: 61 and 63.

[0021] In yet other embodiments, the antigen recognition portion is a single chain variable (scFV) comprising the light chain variable amino acid sequence of SEQ ID NO:49 and the heavy chain variable amino acid sequence of SEQ ID NO:51.

[0022] In some embodiments, the antigen recognition portion is an scFV comprising the light chain variable amino acid sequence of SEQ ID NO:53 and the heavy chain variable amino acid sequence of SEQ ID NO:55.

[0023] In other embodiments, the antigen recognition portion is an scFV comprising the light chain variable amino acid sequence of SEQ ID NO:57 and the heavy chain variable amino acid sequence of SEQ ID NO:59.

[0024] In yet other embodiments, the antigen recognition portion is an scFV comprising the light chain variable amino acid sequence of SEQ ID NO:61 and the heavy chain variable amino acid sequence of SEQ ID NO:63.

[0025] In some embodiments, the antigen recognition portion is an scFV comprising the light chain variable amino acid sequence of SEQ ID NO:65 and the heavy chain variable amino acid sequence of SEQ ID NO:67.

[0026] In other embodiments, the antigen recognition portion is an scFV comprising the light chain variable amino acid sequence of SEQ ID NO:69 and the heavy chain variable amino acid sequence of SEQ ID NO:71.

[0027] In another aspect, provided herein is a heterogeneous T cell population comprising modified and unmodified T cells, wherein the modified T cells comprise a) a first polynucleotide encoding an inducible MyD88 / CD40 polypeptide, and b) a second polynucleotide encoding a PSCA-CAR polypeptide.

[0028] In some embodiments, about 20% to about 90% of the T cells are modified T cells. In some embodiments, about 20% to about 80% of the T cells are modified T cells. In some embodiments, about 20% to about 70% of the T cells are modified T cells. In certain embodiments, about 20% to about 60% of the T cells are modified T cells. In some embodiments, about 20% to about 50% of the T cells are modified T cells.

[0029] In some embodiments, the T cell population is suspended in freezing medium. In certain embodiments, the T cell population provided herein is a cryopreserved T cell population.

[0030] In some embodiments, the T cell populations provided herein comprise about 20×10 T cells per milliliter at a suitable dilution for infusion into a subject. 6 ~150×10 6 In other embodiments, the T cell populations provided herein comprise about 20×10 cells per milliliter at a suitable dilution for infusion into a subject. 6 ~120×10 6 Contains cells of.

[0031] In some embodiments, the T cell populations provided herein are engineered T cells comprising a first polynucleotide encoding an inducible MyD88 / CD40 polypeptide, wherein the inducible MyD88 / CD40 polypeptide is i) two multimeric ligand-binding domains; ii) a MyD88 polypeptide, or a truncated MyD88 polypeptide lacking the TIR domain, and iii) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain The modified T cell comprises:

[0032] In some embodiments, each multimeric ligand binding region comprises an FKBP12 variant polypeptide. In other embodiments, each FKBP12 variant polypeptide binds to a multimeric ligand with higher affinity than wild-type FKBP12 polypeptide. In certain embodiments, each FKBP12 variant polypeptide comprises an amino acid substitution at position 36 and binds to a multimeric ligand with higher affinity than wild-type FKBP12 polypeptide. In some embodiments, the amino acid substitution at position 36 is selected from the group consisting of valine, isoleucine, leucine and alanine. In some embodiments, each multimeric ligand binding region is an FKB12v36 region.

[0033] In some embodiments, the T cell populations provided herein are engineered T cells comprising a first polynucleotide encoding an inducible MyD88 / CD40 polypeptide, wherein the inducible MyD88 / CD40 polypeptide is i) two multimeric ligand-binding domains; ii) a MyD88 polypeptide, or a truncated MyD88 polypeptide lacking the TIR domain, and iii) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain and wherein the truncated MyD88 polypeptide has the amino acid sequence of SEQ ID NO:21, or a functional fragment thereof. In certain embodiments, the truncated MyD88 polypeptide consists of the amino acid sequence of SEQ ID NO:21.

[0034] In some embodiments, the T cell populations provided herein are engineered T cells comprising a first polynucleotide encoding an inducible MyD88 / CD40 polypeptide, wherein the inducible MyD88 / CD40 polypeptide is i) two multimeric ligand-binding domains; ii) a MyD88 polypeptide, or a truncated MyD88 polypeptide lacking the TIR domain, and iii) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain and the CD40 cytoplasmic polypeptide comprises the amino acid sequence of SEQ ID NO: 23, or a functional fragment thereof. In some embodiments, the CD40 cytoplasmic polypeptide consists of the amino acid sequence of SEQ ID NO: 23, or a functional fragment thereof.

[0035] In some embodiments, provided herein is a heterogeneous T cell population comprising modified and unmodified T cells, wherein the modified T cells comprise a) a first polynucleotide encoding an inducible MyD88 / CD40 polypeptide, and b) a second polynucleotide encoding a PSCA-CAR polypeptide, wherein the PSCA-CAR polypeptide comprises an antigen recognition moiety, a transmembrane domain, and a T cell activation molecule. In certain embodiments, the antigen recognition portion is derived from a PSCA antibody selected from the group consisting of 1G8, 2A2, 2H9, 3C5, 3E6, 3G3, 4A10, and A11, where 1G8, 2A2, 2H9, 3C5, 3E6, 3G3, and 4A10 are produced by hybridomas designated HB-12612, HB-12613, HB-12614, HB-12616, HB-12618, HB-12615, and HB-12617, respectively, deposited at the American Type Culture Collection.

[0036] In some embodiments, the antigen recognition portion comprises the complementarity determining regions (CDRs) of the heavy and light chain variable domains of the amino acid sequences selected from the group consisting of SEQ ID NOs: 65, 67, 69 and 71. In other embodiments, the antigen recognition portion comprises the heavy and light chain variable amino acid sequences selected from the group consisting of SEQ ID NOs: 51 and 49, SEQ ID NOs: 55 and 53, SEQ ID NOs: 57 and 59, and SEQ ID NOs: 61 and 63. In some embodiments, the antigen recognition portion is an scFV comprising the light chain variable amino acid sequence of SEQ ID NO: 49 and the heavy chain variable amino acid sequence of SEQ ID NO: 51. In some embodiments, the antigen recognition portion is an scFV comprising the light chain variable amino acid sequence of SEQ ID NO: 53 and the heavy chain variable amino acid sequence of SEQ ID NO: 55. In some embodiments, the antigen recognition portion is an scFV comprising the light chain variable amino acid sequence of SEQ ID NO: 57 and the heavy chain variable amino acid sequence of SEQ ID NO: 59. In some embodiments, the antigen recognition portion is an scFV comprising the light chain variable amino acid sequence of SEQ ID NO: 61 and the heavy chain variable amino acid sequence of SEQ ID NO: 63. In some embodiments, the antigen recognition portion is an scFV that comprises a light chain variable amino acid sequence of SEQ ID NO: 65 and a heavy chain variable amino acid sequence of SEQ ID NO: 67. In some embodiments, the antigen recognition portion is an scFV that comprises a light chain variable amino acid sequence of SEQ ID NO: 69 and a heavy chain variable amino acid sequence of SEQ ID NO: 71.

[0037] The drawings are illustrative of certain embodiments of the present technology and are not limiting. For clarity and ease of illustration, the drawings have not been made to scale and in some instances various aspects may be shown exaggerated or enlarged to facilitate understanding of particular embodiments. [Brief description of the drawings]

[0038] [Figure 1] FIG. 1 shows PSCA expression in tumor tissues.

[0039] [Diagram 2] Figure 2 shows a retroviral vector construct containing an inducible MyD88 / CD40 component and an anti-PSCA CAR component. Q, CD34.

[0040] [Diagram 3] Figure 3 shows an illustration of inducible MyD88 / CD40. MyD88 and CD40 signaling pathways are utilized to generate inducible costimulation. IFN, interferon; iMC, inducible MyD88 / CD40; TIR, Toll / interleukin-1 receptor; TLR, Toll-like receptor.

[0041] [Figure 4] Figure 4 shows an illustration of T cells engineered with inducible costimulation (i.e., inducible MyD88 / CD40 PSCA CAR-T cells). Cell characteristics include potential autologous T cell products; antigen-specific activation by PSCA-CD3ζ CAR; and rimiduside-dependent costimulation by inducible MyD88 / CD40 domains. CAR, chimeric antigen receptor; iMC, inducible MyD88 / CD40; PSCA, prostate stem cell antigen; Rim, rimiduside; TCR, T cell receptor.

[0042] [Figure 5-1] Figures 5A-5B show dose-dependent tumor killing by inducible MyD88 / CD40 PSCA CAR-T cells in HPAC-bearing NSG mice. Figure 5A: Tumor volumes measured over time after injection of non-transduced cells or inducible MyD88 / CD40 PSCA CAR-T cells at the indicated doses. Red arrows indicate rimiduside injection for the applicable dose groups. Figure 5B: Viability after injection of non-transduced cells or inducible MyD88 / CD40 PSCA CAR-T cells at the indicated doses. CID = rimiduside; NT = non-transduced T cells. [Figure 5-2] Same as above.

[0043] [Figure 6] Figure 6 shows data supporting the rationale for prostate stem cell antigen (PSCA) targeting. GPI, glycosylphosphatidylinositol; hACTB, human beta actin; LoQ, limit of quantification.

[0044] [Figure 7] FIG. 7 shows an illustration of the cell dosing study design (i.e., using inducible MyD88 / CD40 PSCA CAR-T cells). Objectives of Part 1 of the study included safety, tolerability, MTD and / or RDE for use in Part 2. MTD, maximum tolerated dose; PSCA, prostate stem cell antigen; RDE, recommended dose expansion.

[0045] [Figure 8] Figure 8 provides a table showing patient demographics and clinical characteristics. *-Cytoxan 1g / m2 by IV infusion on day 3; †PSCA (copies per 106 copies of hACTB) measured at screening. CTX, cyclophosphamide; hACTB, human beta-actin; LD, lymphocyte depletion; PSCA, prostate stem cell antigen; Rim, rimiduside.

[0046] [Figure 9] Figure 9 shows a safety summary. No DLTs, neurotoxicity, or cytokine release syndrome events were observed. Fever was the only treatment-related AE (grade 1-2 on day 0 after infusion of inducible MyD88 / CD40 PSCA CAR-T cells) reported by more than one patient (n=2). Both events resolved within 24-36 hours with supportive care. AE = adverse event; DLT = dose-limiting toxicity.

[0047] [Figure 10] Figure 10 shows inducible MyD88 / CD40 PSCA CAR-T cell expansion and persistence. Observations include: little evidence of lymphodepletion (LD) with cyclophosphamide (CTX)-only regimen (79% ± 25% of cells remained); rapid cell expansion by day 4, but no persistence without rimiduside; and 3- to 20-fold cell expansion within 7 days in 4 patients and cell persistence for >3 weeks in 3 patients with single dose rimiduside. *Vector copy number < limit of quantification. CTX, cyclophosphamide; LD, lymphodepletion; PSCA, prostate stem cell antigen; Rim, rimiduside.

[0048] [Figure 11] Figure 11 shows peripheral cytokine profiles over time. Rim, rimiduside.

[0049] [Figure 12] Figure 12 shows evidence of anti-tumor activity in patients treated with inducible MyD88 / CD40 PSCA CAR-T cells. Scans were performed at week 4 (month 1), week 8 (month 2), and every 8 weeks thereafter (every 2 months) until progression. CA19-9, cancer antigen 19-9; CR, complete response; PD, progressive disease; PR, partial response; PSCA, prostate stem cell antigen; SD, stable disease.

[0050] [Figure 13] Figure 13 shows evidence of anti-tumor activity in patients treated with inducible MyD88 / CD40 PSCA CAR-T cells. PSCA, prostate stem cell antigen.

[0051] [Figure 14] Figure 14 shows time to next treatment after administration of inducible MyD88 / CD40 PSCA CAR-T cells. PD, progressive disease; pseudo-PD, pseudo-progression, progressive disease; SD, stable disease. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0052] Prostate stem cell antigen (PSCA) is a glycosylphosphatidylinositol-anchored cell surface protein with 123 amino acids. PSCA is poorly expressed in normal epithelial cells of the prostate, bladder, kidney, skin, esophagus, stomach, eye, and placenta, but PSCA is upregulated in cancers of the pancreas, prostate, stomach, ovary, and bladder (Figure 1; Abate-Daga). D 2014 Hum Gene Ther 25: 1003-1012). Expression of PSCA positively correlates with advanced clinical stage and metastasis of prostate cancer (Saeki N 2010 Cancer Res 16: 3533-3538). PSCA-Expressing Solid Tumors with Significant Unmet Medical Need Pancreatic cancer

[0053] Pancreatic adenocarcinoma is the fourth leading cause of cancer-related deaths in the United States (Siegel RL 2017 CA Cancer J Clin 67: 7-30). Because of poor survival rates, the demographic distribution of people who die from pancreatic cancer is similar to that of people diagnosed with the disease. It is estimated that 55,440 people will be diagnosed with pancreatic adenocarcinoma in 2018, and approximately 44,330 people will die from pancreatic adenocarcinoma (SEER Pancreatic Cancer 2018, from https: / / seer.cancer.gov / statfacts / html / prost.html, accessed May 14, 2018). Radiologic imaging, liver function tests, and levels of circulating tumor biomarkers (CA 19-9) are the primary tools used for diagnosis and staging.

[0054] Surgical resection is the only potentially curative treatment for managing resectable and borderline resectable pancreatic cancer. However, the disease is often difficult to detect at these early stages, and many patients are only diagnosed when the disease has already progressed to an unresectable state, locally advanced, or with distant metastases. For previously untreated subjects with locally advanced or metastatic disease and good performance status, combination chemotherapy remains the standard of care. Randomized clinical trials have shown a survival advantage for both FOLFIRINOX and gemcitabine in combination with nab-paclitaxel as first-line therapy, but median overall survival remains less than 12 months, and severe treatment-related toxicities (particularly myelosuppression and peripheral neuropathy) are common. These data highlight the need to develop safe and effective second-line therapies beyond the current standard of care, which is limited to single-agent chemotherapy or palliative care (Conroy T 2011 N Engl J Med 364: 1817-1825, Von Hoff DD 2013 N Engl J Med 369: 1691-1703). Prostate cancer

[0055] Prostate cancer is the most common non-cutaneous cancer in men in the United States (Litwin MS 2017 JAMA 317: 2532-2542). In 2018, approximately 165,000 men will be diagnosed with prostate cancer, making it the second leading cause of cancer death in men with an estimated annual mortality rate of 29,000 (Siegel RL 2017 CA Cancer J Clin 67: 7-30). Diagnosis is typically made by ultrasound-guided needle biopsy. Histologic patterns are scored using the Gleason system.

[0056] For men with metastatic prostate cancer, androgen deprivation therapy (ADT) with or without docetaxel is the first-line treatment (Sweeney CJ 2015 N Engl J Med 373:737-746, James ND 2016 Lancet 387: 1163-1177, Litwin MS 2017 JAMA 317: 2532-2542). For patients whose metastatic prostate cancer becomes unresponsive to ADT, other drugs that block the androgen pathway (abiraterone, enzalutamide) may slow disease progression, improve survival, and enhance quality of life (de Bono JS 2011 N Engl J Med 364: 1995-2005, Scher H 2012 N Engl J Med 367: 1187-1197, Ryan CJ 2013 N Engl J Med 368:138-148, Beer TM 2014 N Engl J Med 371: 424-433). Other drugs, such as sipuleucel-T, autologous cell therapy (Kantoff PW2010, Schellhammer PF 2013 Urology 81: 1297-1302), and cabazitaxel (de Bono JS 2011 N Engl J Med 364: 1995-2005), taxanes, may be incorporated into treatment. Other drugs, such as denosumab, zoledronic acid, or radium-223, may be used to protect against bone metastasis and bone-related events (Beer TM 2014 N Engl J Med 371: 424-433, Parker C 2013 N Engl J Med 369: 1659-1660). Although improvements have been made with the advent of these therapies, patients with recurrent metastatic prostate cancer have limited survival. Overall survival after chemotherapy for patients with recurrent disease ranges from 14.0 to 18.4 months. Stomach cancer

[0057] Gastric cancer is the fourth most common cancer and the second leading cause of cancer-related deaths worldwide. There are 950,000 new cases per year. In 2012, an estimated 720,000 patients died from gastric cancer (Ferlay J 2015 Int J Cancer 136: E359-E386). Gastric cancer is divided into gastric adenocarcinoma and gastroesophageal junction adenocarcinoma. Histologically, gastric cancer is divided into two main types: diffuse and intestinal. H. pylori infection is the most important cause of distal gastric cancer (Bornschein J 2010 Dig Dis 28: 609-614).

[0058] Treatment options for gastric cancer depend on the treatment setting and include surgery, radiation therapy, chemotherapy, and more recently, targeted therapy with monoclonal antibodies, and immunotherapy. For patients with metastatic gastric cancer, combination chemotherapy, particularly 5-fluorouracil plus platinum, has been established as the state-of-the-art treatment (Kim NK 1993 Cancer 71: 3813-3818, Ohtsu S 2003 J Clin Oncol 21: 54-59, Van Cutsem E 2006 J Clin Oncol 24: 4991-4997). The primary treatment objectives are to prolong survival, relieve symptoms, and improve quality of life, but all patients will eventually experience disease progression with or without clinically significant treatment-related toxicity. For patients undergoing second-line or later treatment, results from clinical trials in Asia have demonstrated improved survival benefits of chemotherapy compared with best supportive care (Takashima A 2014 Gastric Cancer 17: 522-528). Despite these efficacy benefits, the use of chemotherapy after first-line treatment is often restricted to patients with good performance status, highlighting the importance of developing additional subsequent treatment options that not only improve treatment outcomes and prolong survival, but also maintain the quality of life for all patients with advanced gastric cancer. PSCA-specific CAR-T cells

[0059] PSCA-specific CAR-T cells (e.g., inducible MyD88 / CD40 PSCA CAR-T cells) are PSCA-directed, genetically modified, T cells that bind to PSCA-expressing cells.

[0060] T cells bearing first-generation chimeric antigen receptors (CARs), which contain tumor antigen-specific, single-chain variable fragment (scFv) domains and the T cell receptor (TCR)-associated CD3ζ intracellular signaling molecule, are unable to persist or expand in vivo (Kershaw, MH et al. (2006) Clinical cancer research: an official journal of the American Association for Cancer Research 12: 6106-6115;Pule, MA et al. (2008) Nature Med. 14: 1264-1270;Till, BG et al. (2012) Blood 119: 3940-3950), because tumor cells often lack essential costimulatory molecules required for full T cell activation (Inman, BA, et al. (2007) Current cancer drug targets 7: 15-30). Second-generation CAR-T cells incorporating potent intracellular costimulatory domains such as CD28 or 4-1BB (Carpenito, C. et al. (2009) Proc. Natl. Acad. Sci. USA 106: 3360-3365; Song, DG et al. (2012) Blood 119: 696-706) show improved survival and in vivo expansion after adoptive transfer (Kalos, M. et al. (2011) Science translational medicine 3:95ra73; Porter, DL, et al. (2011) The New Eng. J. Med. 365: 725-733; Brentjens, RJ et al. (2013) Science translational medicine 5:177ra138; Kochenderfer, JN et al. (2015) J. Clin. Oncol. 33(6): 540-549). Although CAR-expressing T cells have shown long-term efficacy for the treatment of some types of cancer, toxicities associated with excessive T cell activation, such as cytokine release syndrome (CRS), remain a concern. In addition, CAR-T cell efficacy is more limited in solid tumors due to poor CAR-T cell survival, activation, and proliferation, likely due to the more pronounced inhibitory effect of the tumor microenvironment (Jena, B., et al. (2014) Current hematologic malignancy reports 9: 50- 56;Dotti, G., et al. (2014) Immunological reviews 257: 107-126).Therefore, strategies that allow the controlled expansion and survival of tumor-targeted T cells would maximize efficacy of treatment while minimizing toxicity.

[0061] The antitumor activity of CAR-T cells is associated with robust CAR-T cell expansion following infusion, often accompanied by toxicity (i.e., severe cytokine release syndrome and neurotoxicity), but patients with poor CAR-T proliferation and persistence show reduced long-term remission rates. Examples presented herein demonstrate that controllable signaling from an inducible chimera stimulatory polypeptide, such as inducible MyD88 / CD40 (iMC), can enhance CAR-T survival, proliferation capacity and antitumor activity.

[0062] T cells (also called T lymphocytes) belong to a group of white blood cells called lymphocytes. Lymphocytes are generally involved in cell-mediated immunity. The "T" in "T cell" refers to a cell that originates from the thymus or whose maturation is influenced by the thymus. T cells can be distinguished from other lymphocyte types, such as B cells and natural killer (NK) cells, by the presence of a cell surface protein known as the T cell receptor. The term "activated T cell," as used herein, refers to a T cell that has been stimulated to generate an immune response (e.g., clonal expansion of activated T cells) by recognition of an antigenic determinant, such as one presented in the context of a class II major histocompatibility (MHC) marker. T cells are activated by the presence of antigenic determinants, cytokines and / or lymphokines, and cluster of differentiation cell surface proteins (e.g., CD3, CD4, CD8, etc., and combinations thereof). Cells that express a cluster of differentiation protein are often said to be "positive" for expression of that protein on the cell surface of T cells (e.g., cells positive for CD3, CD4, or CD8 expression are said to be "positive" for CD3, CD4, or CD8 expression). + , CD4 + or CD8 + The CD3 and CD4 proteins are cell surface receptors or co-receptors that may be directly and / or indirectly involved in signal transduction in T cells.

[0063] T cells express receptors on their surface (i.e., T cell receptors) that recognize antigens presented on the surface of cells. During a normal immune response, binding of these antigens to the T cell receptor in the context of MHC antigen presentation initiates intracellular changes that lead to T cell activation. Chimeric antigen receptors (CARs) are artificial receptors designed to convey antigen specificity to T cells without the need for MHC antigen presentation. CARs contain antigen-specific, transmembrane, and intracellular components that are selected to activate T cells and provide specific immunity. Chimeric antigen receptor-expressing T cells can be used in a variety of therapies, including cancer treatment.

[0064] "Chimeric antigen receptor" or "CAR" refers to a chimeric polypeptide that includes a polypeptide sequence (antigen recognition domain, antigen recognition region, antigen recognition portion, or antigen binding region) that recognizes a target antigen, for example, linked to a transmembrane polypeptide and an intracellular domain polypeptide selected to activate T cells and provide specific immunity. The antigen recognition domain can be any polypeptide or fragment thereof, naturally occurring or synthetic, that binds to an antigen, such as an antibody fragment variable domain. For example, antigen recognition portions include, but are not limited to, polypeptides derived from antibodies, such as single chain variable fragments (scFv), Fab, Fab', F(ab')2, and Fv fragments; polypeptides derived from T cell receptors, such as TCR variable domains; polypeptides derived from pattern recognition receptors; and any ligand or receptor fragment that binds to an extracellular cognate protein.

[0065] "T cell activation molecule" refers to a polypeptide that enhances the activation of T cells when incorporated into T cells expressing chimeric antigen receptors. Examples include, but are not limited to, ITAM-containing, Signal 1-conferring molecules, such as CD3ζ (CD3 zeta; CD247) polypeptides, and Fc receptor gamma, such as Fc epsilon receptor gamma (FcεR1γ) subunits (Haynes, NM, et al. J. Immunol. 166:182-7 (2001)). The intracellular domain can include at least one polypeptide that causes the activation of T cells, such as, but not limited to, CD3ζ.

[0066] In some embodiments, the basic components of a chimeric antigen receptor (CAR) include: The variable heavy chain (VH) and variable light chain (VL) of a tumor-specific monoclonal antibody are fused in frame with the CD3 zeta chain (zeta) from the T cell receptor complex. The VH and VL are generally linked to each other using a flexible glycine-serine linker, and then linked to the transmembrane domain by a spacer (e.g., CD8a stalk or CH2CH3) to extend the scFv away from the cell surface so that the scFv can interact with tumor antigens.

[0067] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first-generation CAR construct that contains the classical CD3ζ cytoplasmic signaling domain together with the single-chain variable region domain (scFV) of a humanized PSCA antibody. In some embodiments, anti-PSCA CARs are engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC).

[0068] In some embodiments, the single chain variable region domain (scFv) may be derived from a PSCA antibody selected from the group consisting of 1G8, 2A2, 2H9, 3C5, 3E6, 3G3 and 4A10, produced by the hybridomas designated HB-12612, HB-12613, HB-12614, HB-12616, HB-12618, HB-12615 and HB-12617, respectively, deposited at the American Type Culture Collection, and those discussed in, for example, U.S. Pat. No. 7,527,786 to Reiter, RE et al., issued May 5, 2009, published as U.S. Patent Application Publication No. 2003 / 0113810A1 on June 19, 2003 (all of these references are hereby incorporated by reference in their entirety).

[0069] In some embodiments, the single chain variable region domain (scFV) may be derived from the variable heavy and variable light chains of a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69) and 1G8(2B3-A11) (SEQ ID NO:71).

[0070] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first generation CAR construct containing a classical CD3ζ cytoplasmic signaling domain together with a single chain variable region domain (scFV) containing the variable heavy and light chains A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51), respectively. In some embodiments, an anti-PSCA CAR containing the variable heavy and light chains A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51) is engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC).

[0071] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first generation CAR construct containing a classical CD3ζ cytoplasmic signaling domain together with a single chain variable region domain (scFV) containing the variable heavy and light chains bm2B3 VL (SEQ ID NO: 53) and bm2B3 VH (SEQ ID NO: 55), respectively. In some embodiments, an anti-PSCA CAR containing the variable heavy and light chains bm2B3 VL (SEQ ID NO: 53) and bm2B3 VH (SEQ ID NO: 55) is engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC).

[0072] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first generation CAR construct containing a classical CD3ζ cytoplasmic signaling domain together with a single chain variable region domain (scFV) containing the variable heavy and light chains HA1-4.121 VH (SEQ ID NO: 57) and HLA1-4.121 VL (SEQ ID NO: 59), respectively. In some embodiments, an anti-PSCA CAR containing the variable heavy and light chains HA1-4.121 VH (SEQ ID NO: 57) and HLA1-4.121 VL (SEQ ID NO: 59) is engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC).

[0073] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first generation CAR construct containing a classical CD3ζ cytoplasmic signaling domain together with a single chain variable region domain (scFV) containing the variable heavy and variable light chains H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63), respectively. In some embodiments, an anti-PSCA CAR containing the variable heavy and variable light chains H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63) is engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC).

[0074] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first generation CAR construct containing a classical CD3ζ cytoplasmic signaling domain together with a single chain variable region domain (scFV) containing the variable heavy and variable light chains 1G8(2B3) (SEQ ID NO:65) and 1G8(2B3-C5) (SEQ ID NO:67), respectively. In some embodiments, an anti-PSCA CAR containing the variable heavy and variable light chains 1G8(2B3) (SEQ ID NO:65) and 1G8(2B3-C5) (SEQ ID NO:67) is engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC).

[0075] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first generation CAR construct containing a classical CD3ζ cytoplasmic signaling domain together with a single chain variable region domain (scFV) containing the variable heavy and variable light chains 1G8(2B3-A2) (SEQ ID NO:69) and 1G8(2B3-A11) (SEQ ID NO:71), respectively. In some embodiments, an anti-PSCA CAR containing the variable heavy and variable light chains 1G8(2B3-A2) (SEQ ID NO:69) and 1G8(2B3-A11) (SEQ ID NO:71) is engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC). co-stimulation

[0076] In some embodiments, compositions and methods are provided herein that include CAR-T cell populations that include inducible chimeric stimulating polypeptides. Inducible chimeric stimulating polypeptides can include one or more costimulatory signaling regions, such as CD27, ICOS, RANK, TRANCE, CD28, 4-1BB, OX40, DAP10, MyD88, or CD40, or, for example, cytoplasmic regions thereof. Inducible chimeric stimulating polypeptides can include one or more suitable costimulatory signaling regions that activate signaling pathways activated by CD27, ICOS, RANK, TRANCE, CD28, 4-1BB, OX40, DAP10, MyD88, or CD40. Inducible chimeric stimulating polypeptides can include any polypeptide that activates the NF-κB pathway, Akt pathway, and / or p38 pathway of tumor necrosis factor receptor (TNFR) family (i.e., CD40, RANK / TRANCE-R, OX40, 4-1BB) and CD28 family members (CD28, ICOS). More than one costimulatory polypeptide or costimulatory polypeptide cytoplasmic region can be expressed in the engineered T cells discussed herein. Costimulation provided by MyD88 and CD40

[0077] In some embodiments, the CAR-T cell populations described herein comprise an inducible chimeric stimulating polypeptide. The inducible chimeric stimulating polypeptide can include one or more costimulatory signaling regions that activate a signaling pathway activated by CD27, ICOS, RANK, TRANCE, CD28, 4-1BB, OX40, DAP10, MyD88, or CD40.

[0078] One of the key functions of second generation CARs is their ability to produce IL-2, which supports T cell survival and growth through activation of the nuclear factor of activated T cells (NFAT) transcription factor by CD3ζ (signal 1) and NF-κB (signal 2) by CD28 or 4-1BB.32. Other molecules that similarly activate NF-κB can also be paired with the CD3ζ chain in the CAR molecule. One approach utilizes T cell costimulatory molecules that were originally developed as adjuvants for dendritic cell (DC) vaccines (Narayanan et al. (2011) J Clin Invest 121:1524-1534; Kemnade et al. (2012) Mol Ther 20(7):1462-1471). Toll-like receptor (TLR) signaling is usually required for full activation or licensing of DCs. In TLR signaling, the cytoplasmic TLR / IL-1 domains (called TIR domains) of TLRs dimerize, which recruits and associates with cytoplasmic adaptor proteins, such as myeloid differentiation primary response protein (MyD88; see SEQ ID NO:29 or 47 for the full-length amino acid sequence, and SEQ ID NO:30 or SEQ ID NO:48 for the nucleotide sequence encoding it).

[0079] In some embodiments, the CAR T cell populations described herein comprise an inducible chimeric stimulatory polypeptide that comprises one or more costimulatory signaling regions that activate a signaling pathway activated by a MyD88 polypeptide, a CD40 polypeptide and / or a MyD88-CD40 fusion chimeric polypeptide.

[0080] MyD88 is a universal adaptor molecule for TLRs and key signaling components of the innate immune system that trigger alarm against foreign invaders and prime immune cell recruitment and activation. MyD88 is a cytoplasmic adaptor protein that plays a central role in innate and adaptive immune responses. The protein functions as an essential signal transducer in the interleukin-1 and Toll-like receptor signaling pathways. These pathways regulate the activation of numerous inflammatory genes. The encoded protein consists of an N-terminal death domain and a C-terminal Toll-interleukin-1 receptor domain. The MyD88 TIR domain can heterodimerize with TLRs and homodimerize with other MyD88 proteins. This dimerization in turn recruits and activates IRAK family kinases through interaction of the amino-terminal death domain (DD) of MyD88 with IRAK kinases, thereby initiating signaling pathways that lead to activation of JNK, p38 MAPK (mitogen-activated protein kinase) and NF-κB (transcription factors that induce expression of cytokine- and chemokine-encoding genes (as well as other genes)). MyD88 acts through IRAK1, IRAK2, IRF7 and TRAF6, resulting in NF-κB activation, cytokine secretion and inflammatory responses. MyD88 also activates IRF1, which then rapidly translocates into the nucleus to mediate efficient transduction of IFN-beta, NOS2 / INOS and IL12A genes. MyD88-mediated signaling in intestinal epithelial cells is crucial for maintaining gut homeostasis and controls expression of the antimicrobial lectin REG3G in the small intestine. TLR signaling also involves members of the tumor necrosis factor receptor (TNFR) family and antigen-sensitized CD4 + It interacts with CD40 ligand (CD154 or CD40L) on T cells and upregulates the expression of CD40.

[0081] CD40 is an important part of the adaptive immune response, helping to activate APCs by association with its cognate CD40L, thus biasing stronger CTL responses. The CD40 / CD154 signaling cascade is a key component in T cell function and B cell-T cell interactions. CD40 signaling proceeds through the formation of CD40 homodimers and interaction with TNFR-associated factors (TRAFs), which are executed by the recruitment of TRAFs to the cytoplasmic domain of CD40, leading to T cell activation with the involvement of several secondary signals, such as the NF-κB, JNK and AKT pathways.

[0082] Besides survival and growth advantages, costimulation based on MyD88 or MyD88-CD40 fusion chimeric polypeptides can also provide additional functions to CAR-modified T cells. MyD88 signaling is essential for both Th1 and Th17 responses and, via IL-1, inhibits CD4 + It acts to render T cells resistant to regulatory T cell (Treg)-driven inhibition (see, e.g., Schenten et al. (2014) Immunity 40:78-90). In addition, CD8 + CD40 signaling through Ras, PI3K and protein kinase C in T cells is a key regulator of CD4 + CD25 + This results in NF-κB-dependent induction of granzymes and perforin, cytotoxic mediators that lyse Treg cells (Martin et al. (2010) J Immunol 184:5510-5518). Thus, MyD88 and CD40 coactivation can render CAR-T cells resistant to the immunosuppressive effects of Treg cells, a function that may be of great importance in the treatment of solid tumors and other types of cancer.

[0083] In immune cells, including T cells, MyD88 and CD40 can act together downstream of transcription factors to upregulate the inflammatory cytokine type I IFN and promote proliferation and survival. In addition to the signaling input from CD3ζ from the CAR, MyD88 / CD40 becomes a potent and pleiotropic costimulatory molecule. In some embodiments, the technology described herein provides CAR T cells that contain an inducible chimeric stimulatory polypeptide that includes one or more costimulatory signaling regions that activate the signaling pathways activated by MyD88, CD40 and / or MyD88-CD40 fusion chimeric polypeptide. Examples of suitable costimulatory signaling regions include, but are not limited to, IRAK-4, IRAK-1, TRAF6, TRAF2, TRAF3, TRAF5, Act, JAK3, or any functional fragment thereof.

[0084] One approach for costimulation of CAR-T cells is to express a fusion protein of the signaling element of MyD88 (called MC). The survival and growth of such cells is enhanced by the activation of NFAT activators by the NFAT transcription factor of CD3ζ (signal 1), which is part of the chimeric antigen receptor, and NF-κB (signal 2) by MyD88 and CD40. Activation of CAR-T cells expressing MC is observed with cytoplasmic MyD88 / CD40 chimeric fusion proteins that lack membrane targeting regions, and with chimeric fusion proteins that include MyD88 / CD40 and a membrane targeting region, such as a myristoylation region. CAR-T cells can co-express an inducible chimeric signaling polypeptide that includes a multimeric ligand binding region, such as FKBP12v36, and a MyD88 polypeptide or a truncated MyD8 polypeptide, or a MyD88-CD40 or a truncated MyD88-CD40 polypeptide (iMC). Cells expressing both iMC and first generation CARs allow full T cell activation requiring recognition of both iMC and tumor by CAR, resulting in IL-2 production, upregulation of CD25 receptor, T cell expansion, and therapeutic efficacy regulated by AP1903 in vivo. In some embodiments, the CAR-T cells comprise a nucleic acid encoding a first polynucleotide encoding an inducible chimeric signaling polypeptide and a second polynucleotide encoding a CAR. In some embodiments, the first polynucleotide is located 5' of the second polynucleotide. In some embodiments, the first polynucleotide is located 3' of the second polynucleotide. In some embodiments, a third polynucleotide encoding a linker polypeptide is located between the first polynucleotide and the second polynucleotide. In some embodiments, the linker polypeptide is a 2A polypeptide that can separate the polypeptides encoded by the first and second polynucleotides during or after translation.

[0085] MyD88 or MyD88 polypeptide refers to the polypeptide product of myeloid differentiation primary response gene 88, including, but not limited to, the human version described as NCBI gene ID 4615. An example of a MyD88 polypeptide is provided as SEQ ID NO: 47. Another example of a MyD88 polypeptide is provided as SEQ ID NO: 29. "Truncated" means that the protein is not full-length, e.g., may lack a domain. For example, a truncated MyD88 is not full-length, e.g., may lack a TIR domain. In some embodiments, a truncated MyD88 polypeptide is encoded by the nucleic acid sequence of SEQ ID NO: 22 and comprises the amino acid sequence of SEQ ID NO: 21. A nucleic acid sequence encoding "truncated MyD88" refers to a nucleic acid sequence that encodes a truncated MyD88 peptide, and the term may also refer to a nucleic acid sequence that includes portions that encode any amino acids added as an artifact of cloning, including any amino acids encoded by linkers. Where a method or construct refers to a truncated MyD88 polypeptide, it will be understood that the method may be used, or the construct may be designed, to refer to another MyD88 polypeptide, such as a full-length MyD88 polypeptide. Where a method or construct refers to a full-length MyD88 polypeptide, it will be understood that the method may be used, or the construct may be designed, to refer to a truncated MyD88 polypeptide.

[0086] A "functional equivalent" or "functional fragment" of a MyD88 polypeptide refers to a truncated MyD88 polypeptide, whether or not it lacks a TIR domain, that, when expressed in a cell, e.g., a T cell, a NK cell, or a NK-T cell, is capable of amplifying a cell-mediated tumor killing response, e.g., a T cell-mediated, NK cell-mediated, or NK-T cell-mediated response, e.g., by activating the NFκB pathway. A truncated MyD88 polypeptide can include, e.g., amino acid residues 1-172 of the full-length MyD88 amino acid sequence, e.g., residues 1-172 of SEQ ID NO:29 or SEQ ID NO:47. In some embodiments, a truncated MyD88 polypeptide can include, e.g., amino acid residues 1-151 or 1-155 of the full-length MyD88 amino acid sequence, e.g., residues 1-151 or 1-155 of SEQ ID NO:29 or SEQ ID NO:47. In some embodiments, the truncated MyD88 polypeptide includes, for example, amino acid residues 1 to 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, or 171 of the full-length MyD88 amino acid sequence, examples of full-length MyD88 amino acid sequences being provided as SEQ ID NO: 29 or SEQ ID NO: 47. In some embodiments, the truncated MyD88 amino acid sequence does not include contiguous amino acid residues 173 to 296 of the full-length MyD88 amino acid sequence. In some embodiments, the truncated MyD88 amino acid sequence does not include contiguous amino acid residues 152 to 296 of the full-length MyD88 amino acid sequence. In some embodiments, the truncated MyD88 amino acid sequence does not include contiguous amino acid residues 156 to 296 of the full-length MyD88 amino acid sequence. In some embodiments, the truncated MyD88 amino acid sequence does not include consecutive amino acid residues 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, or 172 to 296 of the full-length MyD88 amino acid sequence. By "full-length MyD88 amino acid sequence" is meant the full-length MyD88 amino acid sequence, e.g., corresponding to SEQ ID NO:29 or SEQ ID NO:47.In the embodiments provided herein, a cytoplasmic CD40 polypeptide lacking an extracellular domain can be present either upstream or downstream from the MyD88 or truncated MyD88 polypeptide portion.

[0087] The term "chimeric stimulating polypeptide" is interchangeable with "chimeric stimulating molecule," "costimulatory polypeptide," "costimulatory molecule," "chimeric costimulatory molecule," and "chimeric costimulatory polypeptide," as well as grammatical variations thereof.

[0088] The terms "chimeric", "fusion" and "chimeric fusion" are used interchangeably herein in reference to a polypeptide containing two or more proteins (or portions of one or more of two or more proteins) that have been linked to generate a chimeric polypeptide. The two or more proteins (or portions thereof) may be directly linked to each other (wherein a terminal amino acid residue of one protein (or portion thereof) is directly attached to a terminal amino acid residue of the other protein (or portion thereof)) or may be linked via one or more intervening elements (e.g., one or more amino acids that are not part of either protein, such as a linker or adapter, or a non-amino acid polymer). For example, a polypeptide generated from a nucleic acid encoding a fusion of a multimerizing protein (or portion thereof) with another protein (e.g., a DNA-binding protein, a transcriptional activator protein, a proapoptotic protein, or a protein component of an immune cell activation pathway) or portion thereof may be referred to as a chimeric, fusion, or chimeric fusion polypeptide.

[0089] Costimulation in T cells expressing chimeric antigen receptors by MyD88 and CD40 polypeptides, and by chimeric signaling polypeptides comprising a costimulatory polypeptide cytoplasmic signal region, is discussed in U.S. Patent Application Publication No. 2016 / 0058857, and in international patent application publication numbers such as WO2018 / 208849, each of which is incorporated by reference herein in its entirety for all purposes.

[0090] For example, non-limiting examples of chimeric polypeptides useful for inducing cell activation and related methods for inducing CAR-T cell activation, including expression constructs, methods for constructing vectors, and activity or fusion assays, can be found in the following patents and patent applications: U.S. Patent Application Publication No. 2014 / 0286987, International Patent Application Publication No. WO2014 / 151960, U.S. Patent Application Publication No. 2016 / 0046700, International Patent Application Publication No. WO2015 / 123527, U.S. Patent Application Publication No. 2004 / 0209836, U.S. Patent No. 7,404,950, International Patent Application Publication No. WO2004 / 073641, U.S. Patent Application Publication No. 2011 / 0033388, U.S. Patent No. 8,691,210, International Patent Application Publication No. WO2008 No. 9,315,559, International Patent Application Publication No. WO2010 / 33949, U.S. Patent Application Publication No. 2011 / 0287038, International Patent Application Publication No. WO2011 / 130566, U.S. Patent Application Publication No. 2016 / 0175359, International Patent Application Publication No. WO2016 / 036746, International Patent Application Publication No. WO2016 / 100241, U.S. Patent Application Publication No. 2017 / 0166877, International Patent Application Publication No. WO2017 / 106185, and International Patent Application Publication No. WO2018 / 208849, each of which is incorporated by reference herein in its entirety, including all of its text, tables, and figures, for all purposes.

[0091] Inducible chimeric stimulating polypeptides (e.g., inducible MyD88 / CD40(iMC)) may include one or more ligand binding regions, including, for example, one or more FKBP regions. In some embodiments, inducible chimeric stimulating polypeptides (e.g., inducible MyD88 / CD40(iMC)) include one or more FKBP12 polypeptides. In some embodiments, inducible chimeric stimulating polypeptides (e.g., inducible MyD88 / CD40(iMC)) include one or more suitable FKBP12 variant polypeptides, including variant polypeptides that bind to AP1903, or other synthetic homodimerizers, such as, for example, AP20187 or AP2015. Variant polypeptides may include, for example, an FKBP region having an amino acid substitution at position 36 selected from the group consisting of valine, leucine, isoleucine, and alanine (Clackson T, et al. Proc Natl Acad Sci USA (1998) 95:10437-10442). In some embodiments, the inducible chimeric stimulating polypeptide (eg, inducible MyD88 / CD40 (iMC)) comprises one or more FKBP12v36 polypeptides.

[0092] In some embodiments, an inducible chimeric stimulating polypeptide (eg, an inducible MyD88 / CD40 (iMC)) comprises two FKBP12v36 polypeptides.

[0093] AP1903 (CAS index name: 2-piperidinecarboxylic acid, 1-[(2S)-1-oxo-2-(3,4,5-trimethoxyphenyl)butyl]-, 1,2-ethanediylbis[imino(2-oxo-2,1-ethanediyl)oxy-3,1-phenylene[(1R)-3-(3,4-dimethoxyphenyl)propylidene]] ester, also known as rimiduside, * ),2R * [S * [S * [1(R * ),2R *]]]]]-(9CI) CAS Registry Number: 195514-63-7; molecular formula: C78H98N4O20 Molecular weight: 1411.65) is a synthetic molecule that has been proven safe in healthy volunteers (Iuliucci JD, et al. (2001) J Clin Pharmacol. 41:870-879).

[0094] As used herein, the term "pharmacologically or pharmacologically acceptable" refers to molecular entities and compositions that do not produce adverse, allergic or other untoward reactions when administered to animals or humans.

[0095] As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. The use of such media and agents for pharmaceutical active substances is known in the art. Except where any conventional media or agent is incompatible with the vector or cell presented herein, its use in therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the composition. In some embodiments, the subject is a mammal. Engineering of expression constructs

[0096] Expression constructs that express the subject chimeric antigen receptors and inducible chimeric stimulating polypeptides are provided herein.

[0097] As used herein, the term "cDNA" is intended to refer to DNA prepared using messenger RNA (mRNA) as template.The advantage of using cDNA, rather than genomic DNA or DNA polymerized from genomic unprocessed or partially processed RNA template, is that cDNA mainly contains the coding sequence of the corresponding protein.There are times when complete or partial genomic sequence is used, such as when non-coding regions are required for expression on demand, or when antisense strategies should target non-coding regions such as introns.

[0098] As used herein, the term "polypeptide" is defined as a chain of amino acid residues, usually having a defined sequence. As used herein, the term polypeptide may be interchangeable with the term "protein."

[0099] As used herein, the term "expression construct" or "transgene" is defined as any type of genetic construct containing a nucleic acid encoding a gene product, where part or all of the nucleic acid coding sequence can be inserted into a vector to be transcribed. A transcript is translated into a protein, but is not necessarily transcribed. In certain embodiments, expression includes both transcription of a gene and translation of mRNA into a gene product. In other embodiments, expression includes only transcription of a nucleic acid encoding a gene of interest. The term "therapeutic construct" is sometimes used to refer to an expression construct or transgene. An expression construct or transgene can be used, for example, as a therapy to treat a hyperproliferative disease or disorder, such as cancer, and thus the expression construct or transgene is a therapeutic or prophylactic construct. As used herein with respect to a disease, disorder, or condition, the terms "treatment," "treat," "treated," or "treating" refer to prevention and / or therapy.

[0100] As used herein, the term "expression vector" refers to a vector that contains a nucleic acid sequence that codes for at least a portion of a gene product that can be transcribed. In some cases, the RNA molecule is then translated into a protein, polypeptide or peptide. In other cases, these sequences are not translated, for example, in the case of the production of antisense molecules or ribozymes. Expression vectors can contain various control sequences, which refer to the nucleic acid sequences necessary for the transcription and possibly translation of an operably linked coding sequence in a particular host organism. In addition to the control sequences that govern transcription and translation, vectors and expression vectors can contain nucleic acid sequences that perform other functions, which are discussed below.

[0101] In certain examples, the polynucleotide encoding the chimeric antigen receptor is in the same vector as the polynucleotide encoding the second polypeptide, such as a viral vector or a plasmid vector. This second polypeptide can be, for example, a chimeric signaling polypeptide, an inducible costimulatory polypeptide, an inducible chimeric stimulatory polypeptide, or a marker polypeptide, as discussed herein. In these examples, the construct can be designed with one promoter operably linked to a nucleic acid comprising a polynucleotide encoding two polypeptides linked by a 2A polypeptide. In this example, the first and second polypeptides are separated during translation, resulting in two polypeptides, or in the example with leaky 2A, either one or two polypeptides. In other examples, the two polypeptides can be expressed separately from the same vector when each nucleic acid comprising a polynucleotide encoding one of the polypeptides is operably linked to a separate promoter. In yet other instances, one promoter may be operably linked to two polynucleotides, thus directing the production of two separate RNA transcripts and thus two polypeptides; in one instance, the promoter may be bidirectional, with the coding regions present in opposite orientations 5'-3'. Thus, the expression constructs discussed herein may include at least one, or at least two promoters.

[0102] In some embodiments, the nucleic acid construct is contained in a viral vector. In certain embodiments, the viral vector is a retroviral vector. In certain embodiments, the viral vector is an adenoviral vector or a lentiviral vector. It will be understood that in some embodiments, the cell is contacted with the viral vector ex vivo, and in some embodiments, the cell is contacted with the viral vector in vivo. Thus, the expression construct can be inserted into a vector, such as a viral vector or a plasmid. The steps of the provided methods can be performed using any suitable method, including, but not limited to, the transduction, transformation, or otherwise providing nucleic acid to a cell described herein.

[0103] As used herein, the term "gene" is defined as a functional protein-, polypeptide-, or peptide-encoding unit. It will be understood that this functional term includes genomic sequences, cDNA sequences, and smaller engineered gene segments that express, or are configured to express, proteins, polypeptides, domains, peptides, fusion proteins, and / or variants.

[0104] As used herein, the term "polynucleotide" is defined as a chain of nucleotides. Furthermore, a nucleic acid is a polymer of nucleotides. Thus, nucleic acid and polynucleotide are interchangeable as used herein. A nucleic acid is a polynucleotide that can be hydrolyzed into monomeric "nucleotides". Monomeric nucleotides can be hydrolyzed into nucleosides. As used herein, a polynucleotide includes, but is not limited to, any nucleic acid sequence obtained by any means available in the art, including, but not limited to, recombinant means, i.e., cloning a nucleic acid sequence from a recombinant library or a cell genome, using conventional cloning techniques and PCR™ and the like, as well as by synthetic means. Furthermore, a polynucleotide includes mutations of polynucleotides, including, but not limited to, mutations of nucleotides or nucleosides by methods well known in the art. A nucleic acid can include one or more polynucleotides.

[0105] "Function-conserving variants" are proteins or enzymes that have been subjected to exchange of a given amino acid residue without altering the overall conformation and function of the protein or enzyme, including but not limited to the replacement of amino acids with those that have similar properties, including polar or non-polar properties, size, shape and charge. Conservative amino acid substitutions for many of the commonly known amino acids that are not encoded by genes are well known in the art. Conservative substitutions of other non-encoded amino acids can be determined based on their physical properties, compared to the properties of the amino acids encoded by genes.

[0106] Amino acids other than those shown to be conserved may differ in a protein or enzyme, and thus the percent protein or amino acid sequence similarity between any two proteins of similar function may vary, for example, at least 70%, at least 80%, at least 90% and at least 95%, as determined according to an alignment scheme. As referred to herein, "sequence similarity" refers to the degree to which nucleotide or protein sequences are related. The degree of similarity between two sequences may be based on sequence identity and / or percent conservation. "Sequence identity" herein refers to the degree to which two nucleotide or amino acid sequences are invariant. "Sequence alignment" refers to the process of lining up two or more sequences to achieve the maximum level of identity (and conservation in the case of amino acid sequences) for the purpose of assessing the degree of similarity. Numerous methods for assessing similarity / identity for aligning sequences are known in the art, such as the cluster method where similarity is based on the MEGALIGN algorithm, as well as BLASTN, BLASTP and FASTA. When using any of these programs, it is possible to select the settings that give the highest sequence similarity.

[0107] As used herein, the term "promoter" is defined as a DNA sequence that is recognized by the synthetic machinery of a cell or by an introduced synthetic machinery and is required to initiate the specific transcription of a gene. In some embodiments, the promoter is a developmentally regulated promoter. As used herein, the term "under transcriptional control," "operably linked," or "operably linked" is defined as a promoter being in the correct position and orientation relative to a nucleic acid to control the initiation of RNA polymerase and the expression of a gene. In some instances, one or more polypeptides are said to be "operably linked." In general, the term "operably linked" is intended to indicate that a promoter sequence is functionally linked to a second sequence, such that the promoter sequence initiates and mediates the transcription of the DNA corresponding to the second sequence.

[0108] The individual promoter utilized to control the expression of the polynucleotide sequence of interest is not considered to be important, as long as it can direct the expression of the polynucleotide in target cells.For example, when targeting human cells, the polynucleotide sequence coding region can be placed adjacent to, for example, a promoter that can be expressed in human cells, and placed under the control of such a promoter.Generally speaking, such a promoter can include either a human promoter or a viral promoter.A promoter that is suitable for the vector used to express CAR or other polypeptides provided herein can be selected.

[0109] In various embodiments, for example, when the expression vector is a retrovirus, one example of a suitable promoter is the Moloney murine leukemia virus promoter. In other embodiments, the promoter can be, for example, the (CMV) immediate early gene promoter, the SV40 early promoter, the Rous sarcoma virus long terminal repeat, β-actin, the rat insulin promoter, and the glyceraldehyde-3-phosphate dehydrogenase promoter, which can be used to achieve high levels of expression of the coding sequence of interest. The use of other viral or mammalian cell or bacterial phage promoters known in the art to achieve expression of the coding sequence of interest is also contemplated, provided that the expression level is sufficient for a given purpose. By utilizing a promoter with well-known properties, the expression level and expression pattern of the protein of interest after transfection or transformation can be optimized.

[0110] Promoters and other regulatory elements are selected so that they are functional in the desired cells or tissues.In addition, this list of promoters should not be construed as exhaustive or limiting.Other promoters that can be used with the promoters and methods disclosed herein.

[0111] The nucleic acids discussed herein may include one or more polynucleotides. In some embodiments, one or more polynucleotides may be described as being located at the "5" or "3" of another polynucleotide, or as being "at" the "5" or "3", or as being located in "5' to 3' order". References to 5' to 3' in these contexts are understood to refer to the direction of the coding region of the polynucleotide in the nucleic acid, for example, where a first polynucleotide is located 5' of a second polynucleotide and connected to a third polynucleotide that encodes a non-cleavable linker polypeptide, and the translation product results in a polypeptide encoded by the first polynucleotide located at the amino terminus of a larger polypeptide that includes the translation products of the first, third and second polynucleotides.

[0112] In some embodiments, two or more polypeptides, such as, for example, a chimeric antigen receptor, an inducible chimeric stimulating polypeptide (e.g., inducible MyD88 / CD40 (iMC)), and / or an additional polypeptide, can be expressed in a cell using two separate vectors. The vectors can be co-transfected or co-transformed into the cell, or the vectors can be introduced into the cell at different times.

[0113] Polypeptides may vary in their order from amino terminus to carboxy terminus. For example, the order of the vector components presented in FIG. 2 and discussed in Example 1 may vary. In some embodiments, the order of the components of an inducible chimeric stimulating polypeptide may vary. In some embodiments, the order of the components of a chimeric antigen receptor polypeptide may vary. Various domain orders may be assayed to obtain optimal expression and activity, for example using methods such as those discussed herein.

[0114] In some embodiments, when the expression construct encodes a MyD88 polypeptide, the polypeptide may be a portion of a full-length MyD88 polypeptide. By MyD88, or MyD88 polypeptide, is meant the polypeptide product of myeloid differentiation primary response gene 88, such as, but not limited to, the human version described as NCBI gene ID 4615. In some embodiments, the expression construct encodes a portion of a MyD88 polypeptide that lacks the TIR domain. In some embodiments, the expression construct encodes a portion of a MyD88 polypeptide that contains a DD (death domain), or that contains a DD and a middle domain. "Truncated" means that the protein is not full-length, e.g., may lack a domain. For example, a truncated MyD88 may not be full-length, e.g., may lack the TIR domain. In some embodiments, the truncated MyD88 polypeptide has the amino acid sequence of SEQ ID NO: 21, or a functionally equivalent fragment thereof. In some embodiments, the truncated MyD88 polypeptide is encoded by the nucleotide sequence of SEQ ID NO: 22, or a functionally equivalent fragment thereof. A functionally equivalent portion of a MyD88 polypeptide has substantially the same ability to stimulate intracellular signaling as the polypeptide of SEQ ID NO:21 and has at least 50%, 60%, 70%, 80%, 90% or 95% of the activity of the polypeptide of SEQ ID NO:21. In some embodiments, the expression construct encodes a portion of a MyD88 polypeptide that lacks the TIR domain. A nucleic acid sequence encoding a "truncated MyD88" refers to a nucleic acid sequence that encodes a truncated MyD88 polypeptide, which term may also refer to a nucleic acid sequence that includes portions that encode any amino acids added as an artifact of cloning, including any amino acids encoded by linkers.

[0115] It will be understood that when a method or construct refers to a truncated MyD88 polypeptide, the method may be used or the construct may be designed to refer to another MyD88 polypeptide, such as a full-length MyD88 polypeptide. It will be understood that when a method or construct refers to a full-length MyD88 polypeptide, the method may be used or the construct may be designed to refer to a truncated MyD88 polypeptide. In the methods herein, where a chimeric polypeptide comprises a MyD88 polypeptide (or a portion thereof) and a CD40 polypeptide (or a portion thereof), the MyD88 polypeptide of the chimeric polypeptide may be located upstream or downstream from the CD40 polypeptide. In certain embodiments, the MyD88 polypeptide (or a portion thereof) is located upstream of the CD40 polypeptide (or a portion thereof). As used herein, the term "functionally equivalent" refers to, for example, a MyD88 polypeptide that stimulates a cell signaling response, or a nucleic acid encoding such a MyD88 polypeptide, when the term relates to MyD88 or a portion thereof. "Functionally equivalent" refers, for example, to a MyD88 polypeptide that lacks the TIR domain but is capable of stimulating a cell signaling response. Selectable markers

[0116] In certain embodiments, the expression construct contains a nucleic acid construct whose expression is identified in vitro or in vivo by including a marker in the expression construct. Such a marker confers an identifiable change to the cell, which allows easy identification of cells containing the expression construct. Usually, the inclusion of a drug selection marker aids in cloning and selection of transformants. For example, genes that confer resistance to neomycin, puromycin, hygromycin, DHFR, GPT, zeocin, and histidinol are useful selectable markers. Alternatively, enzymes such as herpes simplex virus thymidine kinase (tk) are utilized. Immunological surface markers that contain extracellular non-signaling domains or various proteins (e.g., CD34, CD19, LNGFR) can also be utilized, which allows for easy magnetic or fluorescent antibody-mediated selection methods. The selectable marker is not considered important, so long as it is capable of being expressed simultaneously with the nucleic acid encoding the gene product. Further examples of selectable markers include reporters, such as GFP, EGFP, β-gal or chloramphenicol acetyltransferase (CAT).In certain embodiments, marker proteins, such as CD19, are used to select cells for transfusion, such as by immunomagnetic selection.As discussed herein, CD19 markers are distinct from anti-CD19 antibodies, or from, for example, scFv, TCR, or other antigen recognition moieties that bind to CD19.

[0117] In certain embodiments, the marker polypeptide is linked to the inducible chimeric stimulating polypeptide. For example, the marker polypeptide can be linked to the inducible chimeric stimulating polypeptide via a polypeptide sequence, such as a cleavable 2A-like sequence.

[0118] The CAR-T cells provided herein can express a cell surface transgene marker that is present on an expression vector expressing the CAR, or, in some embodiments, is present on an expression vector encoding a protein other than the CAR that is co-expressed with the CAR, such as an inducible chimeric stimulating polypeptide.

[0119] In one embodiment, the cell surface transgene marker is a truncated CD19 (ΔCD19) polypeptide (Di Stasi et al. (2011) supra), which comprises human CD19 truncated at amino acid 333 to remove most of the cytoplasmic domain. The extracellular CD19 domain can still be recognized (e.g., by flow cytometry, FACS or MACS), but the potential to induce intracellular signaling is minimized. Because CD19 is normally expressed by B cells but not by T cells, selection of CD19+ T cells allows for the separation of genetically modified T cells from non-modified donor T cells.

[0120] In some embodiments, the marker polypeptide can include a polypeptide, for example, a CAR encoded by an expression vector to aid in cell sorting. In some embodiments, the expression vector used to express the chimeric antigen receptor or inducible chimeric stimulating polypeptide provided herein further includes a polynucleotide encoding the 16 amino acid CD34 minimal epitope. In some embodiments, the CD34 minimal epitope is incorporated into the amino terminal portion of the CD8 stalk. Linker Polypeptides

[0121] Linker polypeptides include, for example, cleavable and non-cleavable linker polypeptides. Non-cleavable polypeptides can include, for example, any polypeptide that can be operably linked between a MyD88-CD40 chimeric polypeptide, a MyD88 polypeptide, a CD40 polypeptide, or a costimulatory polypeptide cytoplasmic signaling region of a chimeric antigen receptor and the CD3ζ portion. Linker polypeptides can include, for example, those consisting of about 2 to about 30 amino acids (e.g., a furin cleavage site, (GGGGS) n ). In some embodiments, the linker polypeptide consists of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 amino acids. In some embodiments, the linker polypeptide consists of about 18-22 amino acids. In some embodiments, the linker polypeptide consists of 20 amino acids. In some embodiments, the cleavable linker comprises a linker that is cleaved by an enzyme exogenous to the modified cells in the population, e.g., an enzyme encoded by a polynucleotide introduced into the cells by transfection or transduction, either simultaneously or at a different time as the polynucleotide encoding the linker. In some embodiments, the cleavable linker comprises a linker that is cleaved by an enzyme endogenous to the modified cells in the population, including, e.g., an enzyme naturally expressed in the cells and an enzyme encoded by a polynucleotide native to the cells, e.g., a ribozyme. 2A peptide bond skipping sequence

[0122] 2A-like sequences, or "peptide bond skipping" 2A sequences, are derived from, for example, many different viruses, among them, Thosea asigna. These sequences are also known as "peptide skipping sequences". When this type of sequence is placed in a cistron between two polypeptides that are intended to be separated, the ribosome appears to skip the peptide bond in the case of Thosea asigna sequences, and the bond "PGP" between the carboxy-terminal Gly and Pro amino acids is deleted. This may result in two to three polypeptides remaining, for example, an inducible chimeric stimulating polypeptide and a chimeric antigen receptor, or, for example, a marker polypeptide and an inducible chimeric stimulating polypeptide. When this sequence is used, the polypeptide encoded 5' of the 2A sequence may end with additional amino acids at the carboxy-terminus, including a Gly residue and any upstream residues in the 2A sequence. The peptide encoded 3' of the 2A sequence may end with additional amino acids at the amino-terminus, including a Pro residue and any downstream residues following the 2A sequence. In some embodiments, the cleavable linker is a 2A polypeptide (P2A) from porcine teschovirus-1. In some embodiments, the 2A cotranslational sequence is a 2A-like sequence. In some embodiments, the 2A cotranslational sequence is T2A (Thosea asigna virus 2A), F2A (foot and mouth disease virus 2A), P2A (porcine teschovirus-1 2A), BmCPV 2A (cytoplasmic polyhedrosis virus 2A), BmIFV 2A (B. mori flacherie virus 2A), or E2A (equine rhinitis A virus 2A). In some embodiments, the 2A cotranslational sequence is T2A-GSG, F2A-GSG, P2A-GSG, or E2A-GSG. In some embodiments, the 2A cotranslational sequence is selected from the group consisting of T2A, P2A, and F2A. By "cleavable linker" is meant that the linker is cleaved by any means, including, for example, non-enzymatic means, such as peptide skipping, or enzymatic means. (Donnelly, ML 2001, J. Gen. Virol. 82:1013-25).

[0123] 2A-like sequences can also be "leaky" since parts of the polypeptide are not separated during translation but remain as one long polypeptide after translation. One theory for the cause of leaky linkers is that short 2A sequences may not be able to fold into the required structure (the "2A fold") that promotes ribosome skipping. In these cases, the ribosome may miss the proline peptide bond, thus resulting in a fusion protein. To reduce the level of leakiness and thus the number of fusion proteins formed, a GSG (or similar) linker can be added to the amino-terminal side of the 2A polypeptide, which prevents the secondary structure of the newly translated polypeptide from spontaneously folding and disrupting the "2A fold".

[0124] In certain embodiments, the 2A linker comprises the amino acid sequence of SEQ ID NO: 19. In certain embodiments, the 2A linker further comprises a GSG amino acid sequence at the amino terminus of the polypeptide, and in other embodiments, the 2A linker further comprises a GSGPR amino acid sequence at the amino terminus of the polypeptide. Thus, with respect to a "2A" sequence, the term may refer to the 2A sequence in the examples described herein, or may refer to a 2A sequence as recited herein that further comprises a GSG or GSGPR sequence at the amino terminus of the linker.

[0125] In some embodiments, the linker, e.g., the 2A linker, is cleaved in about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the chimeric antigen receptors, i.e., the chimeric antigen receptor portion is spaced apart from the chimeric MyD88 and CD40, MyD88 polypeptide, CD40 polypeptide, or costimulatory polypeptide cytoplasmic signaling region, e.g., CD28, OX40, or 4-1BB. In other embodiments, the 2A linker is cleaved in about 75%, 80%, 85%, 90%, 95%, 98% or 99% of the chimeric antigen receptors. In some embodiments, the 2A linker is cleaved in about 80%-99% of the chimeric antigen receptors. In other embodiments, the 2A linker is cleaved in about 90% of the chimeric antigen receptors. In some embodiments, a constitutively active chimeric antigen receptor polypeptide is present in the modified cell in which the 2A linker is not cleaved, i.e., a chimeric antigen receptor portion representing about 1, 2, 5, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, or 90% of the chimeric antigen receptor polypeptide is linked to a chimeric MyD88 and CD40, a MyD88 polypeptide, a CD40 polypeptide, or a costimulatory polypeptide cytoplasmic signaling region, such as CD28, OX40, or 4-1BB. In other embodiments, the 2A linker is not cleaved in about 5%, 10%, 15%, 20%, or 25% of the chimeric antigen receptors. In some embodiments, the 2A linker is not cleaved in about 5-20% of the chimeric antigen receptors. In some embodiments, the 2A linker is not cleaved in about 10% of the chimeric antigen receptors. Chimeric Antigen Receptor antigen recognition part

[0126] An "antigen recognition portion" can be any polypeptide or fragment thereof, naturally derived or synthetic, that binds to an antigen, such as an antibody fragment variable domain. Examples of antigen recognition portions include, but are not limited to, polypeptides derived from antigens, such as single chain variable fragments (scFv), Fab, Fab', F(ab')2, and Fv fragments; polypeptides derived from T cell receptors, such as TCR variable domains; secreted factors (e.g., cytokines, growth factors) that can be artificially fused to signaling domains (e.g., "zytokines"), as well as any ligand or receptor fragment that binds to extracellular cognate proteins (e.g., CD27, NKG2D). Combinatorial libraries can also be used to identify peptides that bind to tumor-associated targets with high affinity. In addition, a "universal" CAR can be created by fusing aviden to the signaling domain in combination with a biotinylated tumor targeting antibody, or by using Fc gamma receptor / CD16 to bind to tumors targeted by IgG. transmembrane region

[0127] The chimeric proteins herein may contain single-pass or multiple-pass transmembrane sequences (e.g., at the N-terminus or C-terminus of the chimeric protein). Single-pass transmembrane regions are found in certain CD molecules, tyrosine kinase receptors, serine / threonine kinase receptors, TGFβ, BMP, activins, and phosphatases. Single-pass transmembrane regions often contain a signal peptide region and a transmembrane region of about 20 to about 25 amino acids, many of which are hydrophobic and capable of forming alpha helices. A short track of positively charged amino acids is often followed by a transmembrane span that anchors the protein in the membrane. Multiple-pass proteins include ion pumps, ion channels, and transporters, and contain two or more helices that span the membrane multiple times. All or substantially all of the multiple-pass proteins may be incorporated into the chimeric protein. The sequences of single-pass and multiple-pass transmembrane regions are known and may be selected for incorporation into the chimeric protein molecule.

[0128] In some embodiments, the transmembrane domain is fused to the extracellular domain of the CAR. In one embodiment, a transmembrane domain that is naturally associated with one of the domains in the CAR is used. In other embodiments, a transmembrane domain that is not naturally associated with one of the domains in the CAR is used. In some cases, the transmembrane domain can be selected or modified by amino acid substitution (e.g., typically from charged residues to hydrophobic residues) to avoid such domains binding to the transmembrane domains of the same or different surface membrane proteins, minimizing interaction with other members of the receptor complex.

[0129] The transmembrane domain may be derived from, for example, the alpha, beta or zeta chain of the T cell receptor, CD3-ε, CD3 ζ, CD4, CD5, CD8, CD8α, CD9, CD16, CD22, CD28, CD33, CD38, CD64, CD80, CD86, CD134, CD137 or CD154. Alternatively, in some cases, the transmembrane domain may be synthesized de novo, comprising primarily hydrophobic residues, such as leucine and valine. In certain embodiments, a short polypeptide linker forms the link between the transmembrane domain and the intracellular domain of the chimeric antigen receptor. The chimeric antigen receptor may further comprise a stalk, i.e., an extracellular region of amino acids between the extracellular domain and the transmembrane domain. For example, the stalk may be a sequence of amino acids that are naturally associated with the selected transmembrane domain. In some embodiments, a chimeric antigen receptor comprises a CD8 transmembrane domain, in certain embodiments, a chimeric antigen receptor comprises a CD8 transmembrane domain and additional amino acids on the extracellular portion of the transmembrane domain, in certain embodiments, a chimeric antigen receptor comprises a CD8 transmembrane domain and a CD8 stalk. A chimeric antigen receptor may further comprise a region of amino acids between the transmembrane domain and the cytoplasmic domain that are naturally associated with the polypeptide from which the transmembrane domain is derived.

[0130] In some embodiments, the chimeric antigen receptor is an anti-PSCA chimeric antigen receptor.

[0131] Anti-PSCA chimeric antigen receptors include an antigen recognition portion that binds to PSCA. Examples of antigen recognition portion polypeptides and heavy and light chain variable amino acid sequences that bind to PSCA are described, for example, in U.S. Patent Application Publication No. 2009 / 0181034A1 published on July 16, 2009, and issued on September 6, 2011, by Gudas, J. U.S. Patent Application Publication No. 2009 / 0202548A1 published on August 13, 2009, and issued on June 26, 2012, by Gudas, J. No. 32; U.S. Patent No. 7,541,442, issued June 2, 2009, published on July 6, 2006 as U.S. Patent Application Publication No. 2006 / 0147375A1; and U.S. Patent No. 8,940,298, issued January 27, 2015, by Wu, AW et al., published on November 25, 2010 as U.S. Patent Application Publication No. 2010 / 0297004, all of which are hereby incorporated by reference in their entireties. Exemplary antigen recognition portion polypeptides may be derived from a PSCA antibody selected from the group consisting of 1G8, 2A2, 2H9, 3C5, 3E6, 3G3 and 4A10 produced by hybridomas designated HB-12612, HB-12613, HB-12614, HB-12616, HB-12618, HB-12615 and HB-12617, respectively, deposited at the American Type Culture Collection, and those discussed in U.S. Patent No. 7,527,786, issued May 5, 2009, by Reiter, RE et al., published as U.S. Patent Application Publication No. 2003 / 0113810A1 on June 19, 2003 (all of these references are hereby incorporated by reference in their entireties). "Derived from" means that the antigen recognition portion that binds to PSCA comprises a polypeptide comprising the variable heavy and variable light chain polypeptides of a PSCA antibody produced by the specified hybridoma, or a polypeptide having 70, 80, 90, 95, 97, 98 or 99 percent homology to these polypeptides.

[0132] In some embodiments, the antigen recognition portion polypeptide is a single chain variable region domain (scFV) from a PSCA antibody selected from the group consisting of 1G8, 2A2, 2H9, 3C5, 3E6, 3G3, and 4A10, produced by hybridomas designated HB-12612, HB-12613, HB-12614, HB-12616, HB-12618, HB-12615, and HB-12617, respectively, deposited at the American Type Culture Collection, and those discussed in U.S. Pat. No. 7,527,786, issued May 5, 2009, by Reiter, RE et al., published as U.S. Patent Application Publication No. 2003 / 0113810A1 on June 19, 2003 (all of these references are hereby incorporated by reference in their entireties). In some embodiments, the single chain variable region domain (scFV) is A11 VL (SEQ ID NO: 49), A11 VH (SEQ ID NO: 51); bm2B3 VL (SEQ ID NO: 53), bm2B3 The variable heavy and light chains are derived from a PSCA antibody selected from the group consisting of VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0133] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first generation CAR construct containing a classical CD3ζ cytoplasmic signaling domain together with a single chain variable region domain (scFV) containing the variable heavy and light chains A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51), respectively. In some embodiments, an anti-PSCA CAR containing the variable heavy and light chains A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51) is engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC).

[0134] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first generation CAR construct containing a classical CD3ζ cytoplasmic signaling domain together with a single chain variable region domain (scFV) containing the variable heavy and light chains bm2B3 VL (SEQ ID NO: 53) and bm2B3 VH (SEQ ID NO: 55), respectively. In some embodiments, an anti-PSCA CAR containing the variable heavy and light chains bm2B3 VL (SEQ ID NO: 53) and bm2B3 VH (SEQ ID NO: 55) is engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC).

[0135] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first generation CAR construct containing a classical CD3ζ cytoplasmic signaling domain together with a single chain variable region domain (scFV) containing the variable heavy and light chains HA1-4.121 VH (SEQ ID NO: 57) and HLA1-4.121 VL (SEQ ID NO: 59), respectively. In some embodiments, an anti-PSCA CAR containing the variable heavy and light chains HA1-4.121 VH (SEQ ID NO: 57) and HLA1-4.121 VL (SEQ ID NO: 59) is engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC).

[0136] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first generation CAR construct containing a classical CD3ζ cytoplasmic signaling domain together with a single chain variable region domain (scFV) containing the variable heavy and variable light chains H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63), respectively. In some embodiments, an anti-PSCA CAR containing the variable heavy and variable light chains H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63) is engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC).

[0137] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first generation CAR construct containing a classical CD3ζ cytoplasmic signaling domain together with a single chain variable region domain (scFV) containing the variable heavy and variable light chains 1G8(2B3) (SEQ ID NO:65) and 1G8(2B3-C5) (SEQ ID NO:67), respectively. In some embodiments, an anti-PSCA CAR containing the variable heavy and variable light chains 1G8(2B3) (SEQ ID NO:65) and 1G8(2B3-C5) (SEQ ID NO:67) is engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC).

[0138] In some embodiments, PSCA-specific CAR-T cells target PSCA with a first generation CAR construct containing a classical CD3ζ cytoplasmic signaling domain together with a single chain variable region domain (scFV) containing the variable heavy and variable light chains 1G8(2B3-A2) (SEQ ID NO:69) and 1G8(2B3-A11) (SEQ ID NO:71), respectively. In some embodiments, an anti-PSCA CAR containing the variable heavy and variable light chains 1G8(2B3-A2) (SEQ ID NO:69) and 1G8(2B3-A11) (SEQ ID NO:71) is engineered to express an inducible dual costimulatory domain composed of two FK binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC). Methods for gene transfer / modification of T cells

[0139] In order to mediate the effects of transgene expression in a cell, it is necessary to transfer the expression construct into the cell. Such transfer may utilize viral or non-viral gene transfer methods. This selection leads to a discussion of gene transfer methods and compositions.

[0140] A transformed cell containing an expression vector is produced by introducing the expression vector into a cell. Polynucleotide delivery methods suitable for transforming an organelle, cell, tissue or organism for use in the current methods include virtually any method by which a polynucleotide (e.g., DNA) can be introduced into an organelle, cell, tissue or organism.

[0141] The terms "cell," "cell line," and "cell culture" as used herein may be used interchangeably. All of these terms include their progeny, which are any and all subsequent generations. It will be understood that all progeny may not be identical due to deliberate or inadvertent mutations. As used herein, the term "ex vivo" refers to "outside" the body. The terms "ex vivo" and "in vitro" may be used interchangeably herein.

[0142] The terms "transfection" and "transduction" are interchangeable and refer to the process by which exogenous nucleic acid sequences are introduced into a eukaryotic host. Transfection (or transduction) can be accomplished by any one of a number of means, including electroporation, microinjection, gene gun delivery, retroviral infection, lipofection, superfection, and the like.

[0143] Any suitable method can be used to transfect or transform cells, such as T cells, or administer the nucleotide sequence or composition of the present method.Certain non-limiting examples are provided herein.In some embodiments, the viral vector is SFG-based viral vector, as discussed in Tey et al. (2007) Biol Blood Marrow Transpl 13:913-24 and by Di Stasi et al. (2011) N Engl J Med 365:1673-83 (2011).

[0144] T cells that are genetically modified as disclosed herein are useful for administration to subjects who can benefit from donor lymphocyte administration. These subjects are typically humans, and thus the methods herein are typically performed using human T cells.

[0145] The modified cells may be obtained from a donor or may be cells obtained from a patient, for example, the cells may be autologous, syngeneic or allogeneic. The cells may be used, for example, for regeneration, for example, to replace the function of diseased cells. The cells may also be modified to express heterologous genes, thus delivering biological agents to specific microenvironments, for example, diseased bone marrow or metastatic deposits. "Therapeutic cells" refers to cells used in cell therapy, i.e., cells administered to a subject to treat or prevent a condition or disease.

[0146] For example, as with cells, "obtained or prepared" means that the cells or cell cultures are isolated, purified, or partially purified from a source, which may be, for example, umbilical cord blood, bone marrow, or peripheral blood. These terms may also apply when the original source or cell culture is cultured, the cells are replicated, and the progeny cells are in turn derived from the original source.

[0147] Peripheral Blood: The term "peripheral blood," as used herein, refers to the cellular components of blood (e.g., red blood cells, white blood cells, and platelets) obtained or prepared from the circulating pool of blood and not sequestered within the lymphatic system, spleen, liver, or bone marrow.

[0148] Umbilical cord blood: Umbilical cord blood is distinct from peripheral blood and blood sequestered within the lymphatic system, spleen, liver, or bone marrow. The terms "umbilical cord blood," "umbilical blood," or "cord blood," which may be used interchangeably, refer to the blood that remains in the placenta and in the umbilical cord attached after birth. Umbilical cord blood often contains stem cells, including hematopoietic cells.

[0149] The term "allogeneic" as used herein refers to antigenically distinct HLA or MHC loci between host and donor cells. For example, cells or tissues transferred from the same species may be antigenically distinct. Syngeneic mice may differ at one or more loci (congenic), and allogeneic mice may have the same background. The term "autologous" refers to cells, nucleic acids, proteins, or polypeptides, etc., that are derived from the same individual as the individual to whom they are subsequently administered. The modified cells of the present method may be, for example, autologous cells, such as autologous T cells, etc.

[0150] Donor T cells are generally cultured (usually under activating conditions, e.g., using anti-CD3 and / or anti-CD28 antibodies, optionally with IL-2) before being genetically modified. This step results in a higher cell yield of T cells at the end of the modification process.

[0151] In some embodiments, the sample may or may not be subjected to allogeneic depletion.In the example provided herein, the sample is not subjected to allogeneic depletion, and thus is allogeneic depleted as discussed in Zhou et al.(2015) Blood 125:4103-13.These populations provide a more robust T cell repertoire for providing therapeutic benefits of donor cells.

[0152] T cells can be transduced using viral vectors encoding the polynucleotides of the present application. Suitable transduction techniques include fibronectin fragment CH-296. As an alternative to transduction using viral vectors, cells can be transfected by any suitable method known in the art, for example, with DNA encoding the inducible chimeric stimulating polypeptide of interest and / or chimeric antigen receptor, for example, using calcium phosphate, cationic polymers (e.g., PEI), magnetic beads, electroporation, and commercially available lipid-based reagents, such as Lipofectamine™ and Fugene™. One result of the transduction / transfection step is that various donor T cells are now genetically modified T cells that can express the polypeptide of interest.

[0153] Viral vectors encoding the desired protein may be used in certain embodiments, hi some embodiments, retroviral vectors capable of providing the highest copy number of proviral integrants per cell are used for transduction.

[0154] After transduction / transfection, the cells can be separated from the transduction / transfection material and cultured again to expand the genetically modified T cells. The T cells can be expanded to obtain the desired minimum number of genetically modified T cells.

[0155] The genetically modified T cells can then be separated from the resulting population of cells. Because the inducible chimeric stimulating polypeptide and / or chimeric antigen receptor may not be suitable for positive selection of the desired T cells, in some embodiments, the genetically modified T cells express a cell surface transgene marker of interest. Cells expressing this surface marker can be selected, for example, using immunomagnetic techniques. For example, paramagnetic beads conjugated to monoclonal antibodies that recognize the cell surface transgene marker of interest can be used, for example, using the CliniMACS system (available from Miltenyi Biotec). In some embodiments, no selection step is performed and a mixture of genetically modified and unmodified T cells is administered to the subject.

[0156] In an alternative procedure, the genetically modified T cells are selected, cultured, and then fed after the transduction step. For example, the order of transduction, feeding, and selection can be reversed.

[0157] The result of these procedures is a composition containing genetically modified donor T cells, and thus capable of expressing, for example, an inducible chimeric stimulating polypeptide and / or chimeric antigen receptor of interest (and sometimes a cell surface transgenic marker of interest). These genetically modified T cells can be administered to a recipient, or may be cryopreserved (optionally after further expansion) prior to administration. Treatment

[0158] The modified cell populations provided herein may be used in methods of treating a human subject in need thereof, or may be used to prepare a medicament for treating such a subject. The cells are typically delivered to the recipient subject by injection.

[0159] The terms "patient" or "subject" are interchangeable and as used herein include, but are not limited to, organisms or animals; mammals, including, for example, humans, non-human primates (e.g., monkeys), mice, pigs, cows, goats, rabbits, rats, guinea pigs, hamsters, horses, monkeys, sheep or other non-human mammals; non-mammals, including, for example, non-mammalian vertebrates, such as birds (e.g., chickens or ducks) or fish, and non-mammalian invertebrates. The subject may be, for example, a human, such as a patient who is immunocompromised or suffers from a hyperproliferative disease. The subject may be, for example, a human, such as a patient with cancer and / or solid tumors.

[0160] As used herein, the terms "treatment", "treat", "treated", "treating" refer to prevention and / or therapy. For example, when used in relation to solid tumors, such as cancerous solid tumors, this term refers to prevention by prophylactic treatment, which increases the resistance of the subject against solid tumors or cancer. In some cases, the subject can be treated to prevent familial or genetically related cancers. For example, when used in relation to infectious diseases, this term refers to prophylactic treatment, which increases the resistance of the subject against infection by pathogens, or in other words, reduces the likelihood of the subject being infected by pathogens or showing symptoms of illness caused by said infection, as well as treatment to combat said infection after the subject is infected, for example, to reduce or eliminate said infection or to prevent said infection from worsening.

[0161] A typical dose of T cells for a subject is 10 4 ~10 7 In some embodiments, the dose of T cells for a subject is between about 0.1 x 10 cells / kg. 6 cells / kg ~ approx. 9×10 6 In some embodiments, the dose of T cells for a subject is between about 0.1 x 10 cells / kg. 6 cells / kg ~ approx. 8×10 6In some embodiments, the dose of T cells for a subject is between about 0.1 x 10 cells / kg. 6 cells / kg ~ approx. 7.5×10 6 In some embodiments, the dose of T cells for a subject is between about 0.1 x 10 cells / kg. 6 cells / kg~about 7×10 6 In some embodiments, the dose of T cells for a subject is between about 0.1 x 10 cells / kg. 6 cells / kg~about 6×10 6 In some embodiments, the dose of T cells for a subject is between about 0.3×10 6 cells / kg ~ approx. 5×10 6 In some embodiments, the dose of T cells for a subject is between about 1.25×10 6 cells / kg ~ approx. 2.5×10 6 In some embodiments, the dose of T cells for a subject is between about 0.3×10 6 In some embodiments, the dose of T cells for a subject is about 0.625×10 6 In some embodiments, the dose of T cells for a subject is about 1.25×10 6 In some embodiments, the dose of T cells for a subject is about 2.5×10 6 In some embodiments, the dose of T cells for a subject is about 5×10 6 In some embodiments, the dose of T cells for a subject is about 6×10 6 In some embodiments, the dose of T cells for a subject is about 7×10 6 In some embodiments, the dose of T cells for a subject is about 7.5×10 6 cells / kg (±20%).

[0162] The recipient may be subjected to myeloablative conditioning before being administered the modified cell population comprising genetically modified T cells. As a result, the recipient's own α / β T cells (and B cells) may be depleted before being administered the genetically modified T cells. Similarly, the hematopoietic (stem) cells administered to the recipient may be depleted for α / β cells. In contrast, the genetically modified donor T cells administered to the subject are generally not depleted for α / β cells.

[0163] In some embodiments, a lymphodepleting chemotherapy regimen is administered prior to the infusion of the modified T cells. In some embodiments, the lymphodepleting chemotherapy regimen includes cyclophosphamide 500 mg / m intravenously 5, 4, and 3 days prior to the infusion of the PSCA CAR T cells. 2 and fludarabine 30 mg / m intravenously. 2 unless the combination is not tolerated. If it is determined that combination-based lymphodepletion poses an unfavorable safety risk to the subject, lymphodepletion may proceed with cyclophosphamide alone.

[0164] The recipient can be a child, for example, a child between the ages of 0 and 16, or between the ages of 0 and 10. In some embodiments, the recipient is an adult.

[0165] The subjects who receive the genetically modified T cells may also receive other tissues from an allogeneic donor, for example, they may receive hematopoietic cells and / or hematopoietic stem cells (e.g., CD34+ cells). The allogeneic tissue and the genetically modified T cells are ideally derived from the same donor, and therefore the cells are genetically compatible. In some embodiments, the donor and the recipient are matched unrelated donors, or suitable family members. For example, the donor may be the parent or child of the recipient. Thus, when a subject is identified as needing genetically modified T cells, a suitable donor may be identified as the T cell donor.

[0166] When modified cell populations provided herein, e.g., modified cell populations comprising modified T cells, are used in conjunction with hematopoietic cells and / or hematopoietic stem cells, in some examples, the modified cell population can be administered at a later time point, e.g., 20 to 100 days later.

[0167] In certain embodiments, the method utilizes a chemically induced dimerization (CID) technique to generate conditionally provided costimulation to modified cells. In addition to this technique being inducible, the method is also reversible due to the degradation of the labile dimerizer.

[0168] In some embodiments, the ligand is a small molecule. The ligand is often dimeric, and the ligand may be a dimeric FK506 or a dimeric FK506 analog. In certain embodiments, the ligand is AP1903 (CAS index name: 2-piperidinecarboxylic acid, 1-[(2S)-1-oxo-2-(3,4,5-trimethoxyphenyl)butyl]-, 1,2-ethanediylbis[imino(2-oxo-2,1-ethanediyl)oxy-3,1-phenylene[(1R)-3-(3,4-dimethoxyphenyl)propylidene]] ester, [2S-[1(R * ),2R * [S * [S * [1(R * ),2R * ]]]]]-(9CI) CAS Registry Number: 195514-63-7; Molecular Formula: C78H98N4O20 Molecular Weight: 1411.65). In certain embodiments, the ligand is AP20187. In certain embodiments, the ligand is an AP20187 analog, such as AP1510. In some embodiments, certain analogs are suitable for FKBP12, and certain analogs are suitable for wobble versions of FKBP12. In certain embodiments, one ligand binding region is included in the chimeric protein. In other embodiments, two or more ligand binding regions are included. For example, if the ligand binding region that includes two of these regions is FKBP12, one can be, for example, a wobble version.

[0169] In some embodiments, the dose of rimiduside is between 0.01 mg / kg and 1.0 mg / kg. In some embodiments, a fixed dose of AP1903 is used for injection, which may be, for example, 0.4 mg / kg infused intravenously over 2 hours. The amount of AP1903 required for effective signaling of cells in vitro is about 10-100 nM (MW: 1412 Da). This amount is equivalent to 14-140 μg / L or about 0.014-0.14 mg / kg (1.4-140 μg / kg). Dosages may vary depending on the application, but in certain instances may be in the range of 0.1-10 nM, or in the range of 50-150 nM, 10-200 nM, 75-125 nM, 100-500 nM, 100-600 nM, 100-700 nM, 100-800 nM, or 100-900 nM. Multimeric ligands or CIDs, such as AP1903 (rimiduside), may be administered at, for example, a dose of about 0.01-1 mg / kg of subject body weight, a dose of about 0.05-0.5 mg / kg of subject body weight, 0.1-2 mg / kg of subject body weight, a dose of about 0.05-1.0 mg / kg of subject body weight, a dose of about 0.1-5 mg / kg of subject body weight, a dose of about 0.2-4 mg / kg of subject body weight, a dose of about 0.5-1.0 ... The ligand may be delivered at a dose of about 0.3-3 mg / kg of the subject's body weight, at a dose of about 0.3-2 mg / kg of the subject's body weight, or at a dose of about 0.3-1 mg / kg of the subject's body weight, e.g., about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, or 10 mg / kg of the subject's body weight. In some embodiments, the ligand is provided at a concentration of 0.4 mg / kg per dose, e.g., 5 mg / mL. For example, vials or other containers may be provided containing the ligand in a volume of about 0.25 ml to about 10 ml per vial, e.g., about 0.25, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 ml, e.g., about 2 ml.

[0170] The administration schedule for the ligand (e.g., rimducide) can be determined as appropriate for the patient, and can include, for example, an administration schedule in which the modified T cells are administered at week 0, followed by induction by administration of a chemical inducer of dimerization, followed by administration of additional inducers if necessary to obtain an effective therapeutic result, i.e., at 2, 4, 6, 8, 10, 12, 14, 16, 18, 20 week intervals thereafter, for a total of, for example, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20 weeks.

[0171] The administration schedule of the ligand (e.g., rimducide) can be determined as appropriate for the patient and may include, for example, an administration schedule in which inducible MyD88 / CD40 PSCA CAR-T cells are administered at week 0, followed by induction by administration of a chemical inducer of dimerization, followed by administration of additional inducers if necessary to obtain an effective therapeutic result, i.e., administered at 2, 4, 6, 8, 10, 12, 14, 16, 18, 20 week intervals thereafter, for a total of, for example, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20 weeks.

[0172] In some embodiments, for administration of transduced T cells, one dose of cells is sufficient, followed by multiple doses of ligand. In some embodiments, T cells can be provided more than once. Thus, for example, the administration schedule can be determined to suit the patient, and can include, for example, administration of T cells at week 0, followed by induction by administration of a chemical inducer of dimerization, followed by administration of additional T cells and inducer at two-week intervals for a total of, for example, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 or 30 weeks.

[0173] Other dosing schedules include, for example, a schedule in which one dose of cells and one dose of inducer are administered. In another example, a schedule can include administering cells and inducer at week 0, followed by administration of additional cells and inducer at 4 week intervals for a total of, for example, 4, 8, 12, 16, 20, 24, 28, or 32 weeks.

[0174] In another example, the schedule may include administering the cells once (at week 0) and then administering an inducer (e.g., rimiduside) together at week 0 or after administration of the cells. For example, administration of the cells and the inducer may be performed together, or the cells may be followed by administration of the inducer on the same or next day or 2, 3, 4, 5, or __ days later. In some embodiments, the first administration of the inducer is followed by administration of an additional inducer (e.g., rimiduside) at a certain interval (e.g., 1, 2, 3, or 4 week intervals). In some embodiments, the inducer (rimiduside) is administered to the subject once a day. In some embodiments, the inducer (rimiduside) is administered to the subject three times a week. In some embodiments, the inducer (rimiduside) is administered to the subject twice a week. In some embodiments, the inducer (rimiduside) is administered to the subject once a week. In some embodiments, the inducer (rimiduside) is administered to the subject once every other week. In some embodiments, the inducer (rimiduside) is administered to the subject twice a month. In some embodiments, the inducer (rimiduside) is administered to the subject once a month.

[0175] In another example, a schedule may include administering cells once (at week 0), then administering an inducer (e.g., rimiduside) at week 1, followed by administering additional inducers (rimiduside) at certain intervals (e.g., 1, 2, 3, or 4 week intervals). In some embodiments, the inducer (rimiduside) is administered to the subject once per day. In some embodiments, the inducer (rimiduside) is administered to the subject three times per week. In some embodiments, the inducer (rimiduside) is administered to the subject twice per week. In some embodiments, the inducer (rimiduside) is administered to the subject once per week. In some embodiments, the inducer (rimiduside) is administered to the subject once every other week. In some embodiments, the inducer (rimiduside) is administered to the subject twice per month. In some embodiments, the inducer (rimiduside) is administered to the subject once per month.

[0176] In some embodiments, the dosing schedule for the ligand (e.g., rimducide) includes a dosing schedule in which inducible MyD88 / CD40 PSCA CAR-T cells are administered at week 0, followed by induction by administration of a chemical inducer of dimerization once daily.

[0177] In some embodiments, the dosing schedule for the ligand (e.g., rimducide) comprises a dosing schedule in which inducible MyD88 / CD40 PSCA CAR-T cells are administered at week 0, followed by induction by administration of a chemical inducer of dimerization six times per week.

[0178] In some embodiments, the dosing schedule for the ligand (e.g., rimducide) comprises a dosing schedule in which inducible MyD88 / CD40 PSCA CAR-T cells are administered at week 0, followed by induction by administration of a chemical inducer of dimerization five times a week.

[0179] In some embodiments, the dosing schedule for the ligand (e.g., rimducide) comprises a dosing schedule in which inducible MyD88 / CD40 PSCA CAR-T cells are administered at week 0, followed by induction by administration of a chemical inducer of dimerization four times a week.

[0180] In some embodiments, the dosing schedule for the ligand (e.g., rimducide) comprises a dosing schedule in which inducible MyD88 / CD40 PSCA CAR-T cells are administered at week 0, followed by induction by administration of a chemical inducer of dimerization three times a week.

[0181] In some embodiments, the dosing schedule for the ligand (e.g., rimducide) comprises a dosing schedule in which inducible MyD88 / CD40 PSCA CAR-T cells are administered at week 0, followed by induction by administration of a chemical inducer of dimerization twice weekly.

[0182] In some embodiments, the dosing schedule for the ligand (e.g., rimducide) comprises a dosing schedule in which inducible MyD88 / CD40 PSCA CAR-T cells are administered at week 0, followed by induction by administration of a chemical inducer of dimerization once a week.

[0183] In some embodiments, the dosing schedule for the ligand (e.g., rimducide) comprises a dosing schedule in which inducible MyD88 / CD40 PSCA CAR-T cells are administered at week 0, followed by induction by administration of a chemical inducer of dimerization twice a month.

[0184] In some embodiments, the dosing schedule for the ligand (e.g., rimducide) comprises a dosing schedule in which inducible MyD88 / CD40 PSCA CAR-T cells are administered at week 0, followed by induction by administration of a chemical inducer of dimerization once a month.

[0185] In some embodiments, the subject is administered about 0.1×10 6 cells / kg~9×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose between 100 and 200 cells / kg, followed by rimiduside at intervals of daily, 3 times per week, twice per week, weekly, twice per month, or once per week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice per month, or once per week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once per week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0186] In some embodiments, the subject is administered about 0.1×10 6 cells / kg~8×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose between 100 and 200 cells / kg, followed by rimiduside at intervals of daily, 3 times per week, twice per week, weekly, twice per month, or once per week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice per month, or once per week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once per week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1- 1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0187] In some embodiments, the subject is administered about 0.1×10 6 cells / kg~7.5×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose between 100 and 200 cells / kg, followed by rimiduside at intervals of daily, 3 times per week, twice per week, weekly, twice per month, or once per week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice per month, or once per week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once per week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0188] In some embodiments, the subject is administered about 0.1×10 6 cells / kg~7×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose between 100 and 200 cells / kg, followed by rimiduside at intervals of daily, 3 times per week, twice per week, weekly, twice per month, or once per week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice per month, or once per week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once per week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1- 1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0189] In some embodiments, the subject is administered about 0.1×10 6 cells / kg~6×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose between 100 and 200 cells / kg, followed by rimiduside at intervals of daily, 3 times per week, twice per week, weekly, twice per month, or once per week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice per month, or once per week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once per week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0190] In some embodiments, the subject is administered about 0.3×10 6 cells / kg~5×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose between 100 and 200 cells / kg, followed by rimiduside at intervals of daily, 3 times per week, twice per week, weekly, twice per month, or once per week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice per month, or once per week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once per week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0191] In some embodiments, the subject is administered about 1.25×10 6 cells / kg~2.5×10 6A dose of MyD88 / CD40 PSCA CAR-T cells is administered at a dose of cells / kg, followed by administration of rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0192] In some embodiments, the subject is administered about 0.3×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0193] In some embodiments, the subject is administered about 0.625×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0194] In some embodiments, the subject is administered about 1.25×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0195] In some embodiments, the subject is administered about 2.5×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of 1000 cells / kg (±20%), followed by rimiduside at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the required period (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the required period (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once per week. In some embodiments, the dose of PSCA The CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0196] In some embodiments, the subject is administered about 5×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of 100 mg / kg ± 20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the required period (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the required period (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0197] In some embodiments, the subject is administered about 6×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0198] In some embodiments, the subject is administered about 7×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0199] In some embodiments, the subject is administered about 7.5×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of 100 mg / kg ± 20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the required period (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the required period (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0200] In some embodiments, the subject is administered about 8×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of 100 mg / kg ± 20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the required period (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the required period (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the PSCA CAR-T cells contain scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0201] In some embodiments, the subject is administered about 0.3×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0202] In some embodiments, the subject is administered about 0.625×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0203] In some embodiments, the subject is administered about 1.25×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0204] In some embodiments, the subject is administered about 2.5×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0205] In some embodiments, the subject is administered about 5×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0206] In some embodiments, the subject is administered about 6×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0207] In some embodiments, the subject is administered about 7×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0208] In some embodiments, the subject is administered about 7.5×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0209] In some embodiments, the subject is administered about 8×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0210] In some embodiments, the subject is administered about 0.3×10 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0211] In some embodiments, the subject is administered about 0.625×10 scFVs containing scFVs derived from bm2B3 VL (SEQ ID NO:53) and bm2B3 VH (SEQ ID NO:55). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0212] In some embodiments, the subject is administered about 1.25×10 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0213] In some embodiments, the subject is administered about 2.5×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0214] In some embodiments, the subject is administered about 5×10 scFVs containing scFVs derived from bm2B3 VL (SEQ ID NO:53) and bm2B3 VH (SEQ ID NO:55). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0215] In some embodiments, the subject is administered about 6×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0216] In some embodiments, the subject is administered about 7×10 scFVs containing scFVs derived from bm2B3 VL (SEQ ID NO:53) and bm2B3 VH (SEQ ID NO:55). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0217] In some embodiments, the subject is administered about 7.5×10 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0218] In some embodiments, the subject is administered about 8×10 scFVs containing scFVs derived from bm2B3 VL (SEQ ID NO:53) and bm2B3 VH (SEQ ID NO:55). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0219] In some embodiments, the subject is administered about 0.3×10 scFVs containing HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0220] In some embodiments, the subject is administered about 0.625×10 scFVs containing HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0221] In some embodiments, the subject is administered about 1.25×10 scFVs containing HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0222] In some embodiments, the subject is administered about 2.5×10 scFVs containing the scFVs derived from HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0223] In some embodiments, the subject is administered about 5×10 scFVs containing scFVs derived from HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0224] In some embodiments, the subject is administered about 6×10 scFVs containing HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0225] In some embodiments, the subject is administered about 7×10 scFVs containing HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0226] In some embodiments, the subject is administered about 7.5×10 scFVs containing HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0227] In some embodiments, the subject is administered about 8×10 scFVs containing HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0228] In some embodiments, the subject is administered about 0.3×10 scFVs containing HA1-4.121 VH H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6 MyD88 / CD40 PSCA at a dose of cells / kg (±20%) CAR-T cells are administered, followed by rimiducide at intervals of daily, three times a week, twice a week, weekly, twice a month, or once a week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiducide is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiducide is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiducide is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once a week for the period required (e.g., cancer remission). In some embodiments, the dose of rimiducide is 0.4 mg / kg once a week.

[0229] In some embodiments, the subject is administered about 0.625×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0230] In some embodiments, the subject is administered about 1.25×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0231] In some embodiments, the subject is administered about 2.5×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0232] In some embodiments, the subject is administered about 5×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of 100 mg / kg (±20%) of cells / kg, followed by rimiducid at daily, 3 times per week, twice per week, weekly, twice per month, or once per month intervals for the required period (e.g., cancer remission). In some embodiments, the dose of rimiducid is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiducid is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month or once a month for as long as needed (e.g., cancer remission).In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0233] In some embodiments, the subject is administered about 6×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0234] In some embodiments, the subject is administered about 7×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0235] In some embodiments, the subject is administered about 7.5×10 scFVs containing H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0236] In some embodiments, the subject is administered about 8×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0237] In some embodiments, the subject is administered about 0.3×10 mAbs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0238] In some embodiments, the subject is administered about 0.625×10 scFVs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0239] In some embodiments, the subject is administered about 1.25×10 scFVs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0240] In some embodiments, the subject is administered about 2.5×10 mAbs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0241] In some embodiments, the subject is administered about 5×10 scFVs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0242] In some embodiments, the subject is administered about 6×10 IgG1-derived scFVs containing 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0243] In some embodiments, the subject is administered about 7×10 scFVs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0244] In some embodiments, the subject is administered about 7.5×10 scFVs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0245] In some embodiments, the subject is administered about 8×10 scFVs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0246] In some embodiments, the subject is administered about 0.3×10 mAbs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0247] In some embodiments, the subject is administered about 0.625×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0248] In some embodiments, the subject is administered about 1.25×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0249] In some embodiments, the subject is administered about 2.5×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0250] In some embodiments, the subject is administered about 5×10 mAbs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0251] In some embodiments, the subject is administered about 6×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0252] In some embodiments, the subject is administered about 7×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0253] In some embodiments, the subject is administered about 7.5×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0254] In some embodiments, the subject is administered about 8×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6 MyD88 / CD40 PSCA CAR-T cells are administered at a dose of cells / kg (±20%), followed by rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week.

[0255] In some embodiments, the modified T cells are administered together with unmodified polyclonal T cells. 6 cells / kg ~ approx. 30×10 6 Polyclonal T cells can be administered in amounts of 0.1 x 10 cells / kg. 6 , 0.5×10 6 , 1×10 6, 1.25×10 6 , 2×10 6 , 3×10 6 , 4×10 6 , 5×10 6 , 10×10 6 , 15×10 6 , 20×10 6 , 25×10 6 or 30×10 6 Cells / kg can be dosed.

[0256] In some embodiments, the subject is administered about 0.1×10 6 cells / kg ~ approx. 9×10 6 The modified T cells were administered at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0257] In some embodiments, the subject is administered about 0.1×10 6 cells / kg ~ approx. 8×10 6 The modified T cells were administered at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0258] In some embodiments, the subject is administered about 0.1×10 6 cells / kg ~ approx. 7.5×10 6 The modified T cells were administered at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0259] In some embodiments, the subject is administered about 0.1×10 6 cells / kg~about 7×10 6 The modified T cells were administered at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0260] In some embodiments, the subject is administered about 0.1×10 6 cells / kg~about 6×10 6 The modified T cells were administered at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0261] In some embodiments, the subject is administered about 0.3×10 6 cells / kg ~ approx. 5×10 6 The modified T cells were administered at a dose of 0.1 × 10 6 cells / kg ~ approx. 25×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0262] In some embodiments, the subject is administered about 1.25×10 6 cells / kg ~ approx. 2.5×10 6 The modified T cells were administered at a dose of 0.1 × 10 6 cells / kg ~ approx. 20×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0263] In some embodiments, the subject is administered about 1.25×10 6 Modified T cells at a dose of 0.1 × 10 cells / kg (±20%) 6 cells / kg~20×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0264] In some embodiments, the subject is administered about 2.5×10 6 Modified T cells at a dose of 0.1 × 10 cells / kg (±20%) 6 cells / kg ~ approx. 25×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the modified T cells are A11 VL (SEQ ID NO: 49), A11 VH (SEQ ID NO: 51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO: 57), HLA1-4.121 VL (SEQ ID NO: 59); H1-1.10 VH (SEQ ID NO: 61), H1-1.10 VL (SEQ ID NO: 63); 1G8(2B3) (SEQ ID NO: 65), 1G8(2B3-C5) (SEQ ID NO: 67); 1G8(2B3-A2) (SEQ ID NO: 69), and 1G8(2B3-A11) (SEQ ID NO: 71).

[0265] In some embodiments, the subject is administered about 5×10 6 Modified T cells at a dose of 0.1 × 10 cells / kg (±20%) 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0266] In some embodiments, the subject is administered about 6×10 6 Modified T cells at a dose of 0.1 × 10 cells / kg (±20%) 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0267] In some embodiments, the subject is administered about 7×10 6 Modified T cells at a dose of 0.1 × 10 cells / kg (±20%) 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0268] In some embodiments, the subject is administered about 7.5×10 6 Modified T cells at a dose of 0.1 × 10 cells / kg (±20%) 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the modified T cells are A11 VL (SEQ ID NO: 49), A11 VH (SEQ ID NO: 51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO: 57), HLA1-4.121 VL (SEQ ID NO: 59); H1-1.10 VH (SEQ ID NO: 61), H1-1.10 VL (SEQ ID NO: 63); 1G8(2B3) (SEQ ID NO: 65), 1G8(2B3-C5) (SEQ ID NO: 67); 1G8(2B3-A2) (SEQ ID NO: 69), and 1G8(2B3-A11) (SEQ ID NO: 71).

[0269] In some embodiments, the subject is administered about 8×10 6 Modified T cells at a dose of 0.1 × 10 cells / kg (±20%) 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0270] In some embodiments, the subject is administered about 1.25×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg~20×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0271] In some embodiments, the subject is administered about 2.5×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 25×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0272] In some embodiments, the subject is administered about 5×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0273] In some embodiments, the subject is administered about 6×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0274] In some embodiments, the subject is administered about 7×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0275] In some embodiments, the subject is administered about 7.5×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0276] In some embodiments, the subject is administered about 8×10 scFVs containing scFVs derived from A11 VL (SEQ ID NO: 49) and A11 VH (SEQ ID NO: 51). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0277] In some embodiments, the subject is administered about 1.25×10 6MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg~20×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0278] In some embodiments, the subject is administered about 2.5×10 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg~25×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0279] In some embodiments, the subject is administered about 5×10 scFVs containing scFVs derived from bm2B3 VL (SEQ ID NO:53) and bm2B3 VH (SEQ ID NO:55). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0280] In some embodiments, the subject is administered about 6×10 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0281] In some embodiments, the subject is administered about 7×10 scFVs containing scFVs derived from bm2B3 VL (SEQ ID NO:53) and bm2B3 VH (SEQ ID NO:55). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0282] In some embodiments, the subject is administered about 7.5×10 6MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0283] In some embodiments, the subject is administered about 8×10 scFVs containing scFVs derived from bm2B3 VL (SEQ ID NO:53) and bm2B3 VH (SEQ ID NO:55). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0284] In some embodiments, the subject is administered about 1.25×10 scFVs containing HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg~20×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0285] In some embodiments, the subject is administered about 2.5×10 scFVs containing the scFVs derived from HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 25×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0286] In some embodiments, the subject is administered about 5×10 scFVs containing scFVs derived from HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0287] In some embodiments, the subject is administered about 6×10 scFVs containing HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0288] In some embodiments, the subject is administered about 7×10 scFVs containing HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0289] In some embodiments, the subject is administered about 7.5×10 scFVs containing HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0290] In some embodiments, the subject is administered about 8×10 scFVs containing HA1-4.121 VH (SEQ ID NO:57) and HLA1-4.121 VL (SEQ ID NO:59). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0291] In some embodiments, the subject is administered about 1.25×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg~20×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0292] In some embodiments, the subject is administered about 2.5×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 25×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0293] In some embodiments, the subject is administered about 5×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0294] In some embodiments, the subject is administered about 6×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0295] In some embodiments, the subject is administered about 7×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0296] In some embodiments, the subject is administered about 7.5×10 scFVs containing H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0297] In some embodiments, the subject is administered about 8×10 scFVs containing scFVs derived from H1-1.10 VH (SEQ ID NO: 61) and H1-1.10 VL (SEQ ID NO: 63). 6MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0298] In some embodiments, the subject is administered about 1.25×10 scFVs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 20×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0299] In some embodiments, the subject is administered about 2.5×10 mAbs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 25×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0300] In some embodiments, the subject is administered about 5×10 scFVs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0301] In some embodiments, the subject is administered about 6×10 IgG1-derived scFVs containing 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0302] In some embodiments, the subject is administered about 7×10 scFVs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0303] In some embodiments, the subject is administered about 7.5×10 scFVs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0304] In some embodiments, the subject is administered about 8×10 scFVs containing scFVs derived from 1G8(2B3) (SEQ ID NO: 65) and 1G8(2B3-C5) (SEQ ID NO: 67). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0305] In some embodiments, the subject is administered about 1.25×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg~20×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0306] In some embodiments, the subject is administered about 2.5×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 25×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0307] In some embodiments, the subject is administered about 5×10 mAbs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0308] In some embodiments, the subject is administered about 6×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0309] In some embodiments, the subject is administered about 7×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0310] In some embodiments, the subject is administered about 7.5×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0311] In some embodiments, the subject is administered about 8×10 scFVs containing scFVs derived from 1G8 (2B3-A2) (SEQ ID NO: 69) and 1G8 (2B3-A11) (SEQ ID NO: 71). 6 MyD88 / CD40 PSCA CAR-T cells at a dose of 0.1 × 10 6 cells / kg ~ approx. 30×10 6 The T cells are administered in combination with unmodified polyclonal T cells in an amount between 1000 and 10000 cells / kg. Following T cell administration, rimiduside is administered at intervals of daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, 3 times per week, twice per week, weekly, twice monthly or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once weekly.

[0312] The methods also include methods of treatment or prevention of diseases caused by hyperproliferative diseases.

[0313] Cancers, including solid tumors, that can be treated using the pharmaceutical compositions include, but are not limited to, prostate cancer, pancreatic cancer, gastric cancer, ovarian cancer, bladder cancer, lung cancer, esophageal cancer, endometrial cancer, cervical cancer, or breast cancer.

[0314] In some embodiments, the present invention provides for the treatment or prevention of prostate cancer, pancreatic cancer and gastric cancer.

[0315] In some embodiments, the present invention provides a method for administering to a subject a dose of about 0.1×10 6 cells / kg~8×10 6 The present invention provides for the treatment or prevention of prostate cancer by administering modified T cells at a dose between 1×10 cells / kg, followed by administration of rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the modified T cells are administered at a dose between 0.1×10 6 cells / kg~30×10 6In some embodiments, the modified T cells are administered in combination with unmodified polyclonal T cells in an amount between 100 and 1500 cells / kg. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0316] In some embodiments, the present invention provides a method for administering to a subject a dose of about 5×10 6 The present invention provides for the treatment or prevention of prostate cancer by administering modified T cells at a dose of 0.1×10 cells / kg (±20%), followed by administration of rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, modified T cells are administered at a dose of 0.1×10 6 cells / kg ~ approx. 30×10 6 In some embodiments, the modified T cells are administered in combination with unmodified polyclonal T cells in an amount between 100 and 150 cells / kg. ... VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO: 57), HLA1-4.121 VL (SEQ ID NO: 59); H1-1.10 VH (SEQ ID NO: 61), H1-1.10 VL (SEQ ID NO: 63); 1G8(2B3) (SEQ ID NO: 65), 1G8(2B3-C5) (SEQ ID NO: 67); 1G8(2B3-A2) (SEQ ID NO: 69), and 1G8(2B3-A11) (SEQ ID NO: 71).

[0317] In some embodiments, the present invention provides a method for administering to a subject a dose of about 7×10 6 The present invention provides for the treatment or prevention of prostate cancer by administering modified T cells at a dose of 0.1×10 cells / kg (±20%), followed by administration of rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, modified T cells are administered at a dose of 0.1×10 6 cells / kg ~ approx. 30×10 6 In some embodiments, the modified T cells are administered in combination with unmodified polyclonal T cells in an amount between 100 and 150 cells / kg. ... VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO: 57), HLA1-4.121 VL (SEQ ID NO: 59); H1-1.10 VH (SEQ ID NO: 61), H1-1.10 VL (SEQ ID NO: 63); 1G8(2B3) (SEQ ID NO: 65), 1G8(2B3-C5) (SEQ ID NO: 67); 1G8(2B3-A2) (SEQ ID NO: 69), and 1G8(2B3-A11) (SEQ ID NO: 71).

[0318] In some embodiments, the present invention provides a method for administering to a subject a dose of about 0.1×10 6 cells / kg~8×10 6 The present invention provides for the treatment or prevention of pancreatic cancer by administering modified T cells at a dose between 1×10 cells / kg, followed by administration of rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the modified T cells are administered at a dose of 0.1×10 6 cells / kg ~ approx. 30×10 6The antibody will be administered in combination with unmodified polyclonal T cells in an amount between 100 and 150 cells / kg. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0319] In some embodiments, the present invention provides a method for administering to a subject a dose of about 7×10 6 The present invention provides for the treatment or prevention of pancreatic cancer by administering modified T cells at a dose of 0.1×10 cells / kg (±20%), followed by administration of rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, modified T cells are administered at a dose of 0.1×10 6 cells / kg ~ approx. 30×10 6The antibody will be administered in combination with unmodified polyclonal T cells in an amount between 100 and 150 cells / kg. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0320] In some embodiments, the present invention provides a method for administering to a subject a dose of about 5×10 6 The present invention provides for the treatment or prevention of pancreatic cancer by administering modified T cells at a dose of 0.1×10 cells / kg (±20%), followed by administration of rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, modified T cells are administered at a dose of 0.1×10 6 cells / kg ~ approx. 30×10 6The antibody will be administered in combination with unmodified polyclonal T cells in an amount between 100 and 150 cells / kg. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0321] In some embodiments, the present invention provides a method for administering to a subject a dose of about 0.1×10 6 cells / kg~8×10 6 The present invention provides for the treatment or prevention of gastric cancer by administering modified T cells at a dose between 1×10 cells / kg, followed by administration of rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, the modified T cells are administered at a dose between 0.1×10 6 cells / kg ~ approx. 30×10 6The antibody will be administered in combination with unmodified polyclonal T cells in an amount between 100 and 150 cells / kg. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0322] In some embodiments, the present invention provides a method for administering to a subject a dose of about 7×10 6 The present invention provides for the treatment or prevention of gastric cancer by administering modified T cells at a dose of 0.1×10 cells / kg (±20%), followed by administration of rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, modified T cells are administered at a dose of 0.1×10 6 cells / kg ~ approx. 30×10 6The antibody will be administered in combination with unmodified polyclonal T cells in an amount between 100 and 150 cells / kg. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0323] In some embodiments, the present invention provides a method for administering to a subject a dose of about 5×10 6 The present invention provides for the treatment or prevention of gastric cancer by administering modified T cells at a dose of 0.1×10 cells / kg (±20%), followed by administration of rimiduside at intervals of daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.01-1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, or 1.0 mg / kg. In some embodiments, the dose of rimiduside is 0.4 mg / kg daily, three times a week, twice a week, weekly, twice a month, or once for the period required (e.g., cancer remission). In some embodiments, the dose of rimiduside is 0.4 mg / kg once a week. In some embodiments, modified T cells are administered at a dose of 0.1×10 6 cells / kg ~ approx. 30×10 6The antibody will be administered in combination with unmodified polyclonal T cells in an amount between 100 and 150 cells / kg. In some embodiments, the modified T cells are MyD88 / CD40 PSCA CAR-T cells containing scFVs derived from the variable heavy and variable light chains from a PSCA antibody selected from the group consisting of A11 VL (SEQ ID NO:49), A11 VH (SEQ ID NO:51); bm2B3 VL (SEQ ID NO:53), bm2B3 VH (SEQ ID NO:55); HA1-4.121 VH (SEQ ID NO:57), HLA1-4.121 VL (SEQ ID NO:59); H1-1.10 VH (SEQ ID NO:61), H1-1.10 VL (SEQ ID NO:63); 1G8(2B3) (SEQ ID NO:65), 1G8(2B3-C5) (SEQ ID NO:67); 1G8(2B3-A2) (SEQ ID NO:69), and 1G8(2B3-A11) (SEQ ID NO:71).

[0324] In some embodiments, the copy number of tumor marker in the subject is monitored. In some embodiments, the reduction of the copy number of tumor marker is measured. For example, this reduction can include about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% reduction of the copy number of tumor marker. In some embodiments, the copy number of prostate stem cell antigen (PSCA) in the subject is monitored. In some embodiments, the copy number of prostate stem cell antigen (PSCA) is measured. For example, this reduction can include about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% reduction of the copy number of prostate stem cell antigen (PSCA).

[0325] In some embodiments, the tumor burden in the subject is monitored. Tumor burden may refer to the number of cancer cells, the size of the tumor, and / or the amount of cancer in the body. Tumor burden may also refer to tumor burden. In some embodiments, the reduction of tumor burden is measured. For example, this reduction may include about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% reduction of tumor burden.

[0326] "Reducing tumor size" or "inhibiting tumor growth" or "reducing tumor burden" of a solid tumor refers to a response to treatment or stabilization of the disease according to standard guidelines, such as, for example, Response Evaluation Criteria in Solid Tumors (RECIST) criteria. For example, the reduction can include about a 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% reduction in the diameter of a solid tumor, or about a 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% reduction in the number of tumors, circulating tumor cells, or tumor markers. The size of the tumor can be analyzed by any method, including, for example, a CT scan; an MRI, e.g., CT-MRI; a chest X-ray (for lung tumors); or molecular imaging, e.g., a PET scan, such as a PET scan after administration of an iodine-123 labeled PSA, e.g., a PSMA ligand, such as, for example, where the inhibitor is TROFEX™ / MIP-1072 / 1095; or molecular imaging, e.g., a SPECT, or a PET scan using a PSA, e.g., a PSMA antibody, e.g., capromad pendetide (Prostascint), a 111-iridium labeled PSMA antibody, and the like.

[0327] "Reducing, slowing or inhibiting tumor angiogenesis" means about a 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% reduction in tumor angiogenesis, or about a 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% reduction in the appearance of new vasculature compared to the amount of tumor angiogenesis before treatment. This reduction may refer to a single tumor or may be the total or average angiogenesis in more than one tumor. Methods of measuring tumor angiogenesis include, for example, CAT scan, MRI, e.g., CT-MRI, or molecular imaging, e.g., SPECT, or PET scan, e.g., PET scan after administration of iodine-123 labeled PSA, e.g., a PSMA ligand, such as, for example, where the inhibitor is TROFEX™ / MIP-1072 / 1095, or a PET scan using a PSA, e.g., a PSMA antibody, e.g., capromab pentetide (Prostascint), an indium-111 labeled PSMA antibody, and the like.

[0328] In some embodiments, the progression of the cancer (e.g., prostate cancer, pancreatic cancer, gastric cancer, ovarian cancer) is prevented or delayed for at least 6 months. In some embodiments, the progression of the cancer is prevented or delayed for at least 12 months. In some embodiments, the cancer (e.g., prostate cancer) has a Gleason score of 7, 8, 9, 10 or greater. In some embodiments, the subject exhibits a partial or complete response up to 3 months after administration of the inducible MyD88 / CD40 PSCA CAR-T cells. In some embodiments, the subject exhibits a partial or complete response up to 3 months after administration of the CID (e.g., rimducide). In some embodiments, the subject exhibits a partial or complete response up to 6 months after administration of the inducible MyD88 / CD40 PSCA CAR-T cells. In some embodiments, the subject exhibits a partial or complete response up to 6 months after administration of the CID (e.g., rimducide). In some embodiments, the subject exhibits a partial or complete response up to 9 months after administration of the inducible MyD88 / CD40 PSCA CAR-T cells. In some embodiments, the subject exhibits a partial or complete response up to 9 months after administration of the CID (eg, rimducide).

[0329] If a tumor is present in an organ, or originates from or metastasizes from another tumor in an organ, or produces one or more organ-specific tumor markers, the tumor is classified and named as part of the organ (e.g., prostate, pancreas, stomach, ovary).For example, if a tumor is determined to have the same or similar chromosomal breakpoint as the tumor in the target organ, the tumor is metastasized from the tumor in the organ.In certain cases, tumor (e.g., prostate tumor, pancreas tumor, stomach tumor, ovarian tumor) expresses prostate stem cell antigen (PSCA).

[0330] For example, if a tumor is present in prostate gland, or originates or metastasizes from another tumor in prostate gland, or produces PSA, it will be classified and named as a part of organ, such as prostate cancer tumor.For example, if it is determined that tumor has the same or similar chromosomal breakpoint as the tumor in the prostate gland of the subject, tumor is metastasized from the tumor in prostate gland.

[0331] For example, the indication to adjust or maintain the subsequent dose of the modified T cell and / or the subsequent dose of the drug, such as the subsequent dose of CID (e.g., rimducide), can be provided in any conventional manner. In some embodiments, the indication can be provided in tabular form (e.g., in a physical or electronic medium). For example, the size of the tumor or the number or level of tumor cells in the sample can be provided in a table, and the clinician can compare the symptoms with the list or table of disease stages. The clinician can then identify the indication for the subsequent drug dose from the table. In certain embodiments, the indication can be presented (e.g., displayed) by the computer after the symptoms are provided to the computer (e.g., entered into the computer's memory). For example, this information can be provided to the computer (e.g., entered into the computer by a user or sent to the computer via a remote device in a computer network), and the computer's software can generate an indication to adjust or maintain the subsequent drug dose and / or provide the subsequent drug dose.

[0332] Once the subsequent dose is determined based on the indication, the clinician can administer the subsequent dose or can instruct another person or entity to adjust the dose. The term "clinician" as used herein refers to a decision maker, which in certain embodiments is a medical professional. The decision maker can in some embodiments be a computer or a displayed computer program output, and the medical service provider can act according to the indication or subsequent drug dose displayed by the computer. The decision maker can administer the subsequent dose directly (e.g., inject the subsequent dose into the subject) or can administer remotely (e.g., pump parameters can be changed remotely by the decision maker).

[0333] Treatment of solid tumor cancers (including, for example, prostate, pancreatic, gastric, and ovarian cancers) can be optimized by determining the concentration of tumor-associated biomarkers (e.g., PSCA) over the course of treatment. Patients may respond differently to the course of treatment, so the response to treatment can be monitored by tracking the biomarker concentration or level in various body fluids or tissues. Determining the concentration, level, or amount of a biomarker polypeptide can include determining the full-length polypeptide or a fragment or variant thereof. The fragment or variant is sufficient to be detected, for example, by immunological methods, mass spectrometry, nucleic acid hybridization, and the like. Optimizing treatment for an individual patient can help avoid side effects as a result of overdosing, help determine when treatment is ineffective and change the course of treatment, or help determine when the dose can be increased, or help determine the timing of treatment.

[0334] For example, it has been determined that the amount or concentration of a certain biomarker (e.g., PSCA) changes during the course of treatment of solid tumors. A predetermined target level of such biomarker, or a biomarker threshold that can be identified in normal subjects, is provided, allowing clinicians to determine whether the subsequent dose of drug administered to a subject in need thereof, for example, a subject with a solid tumor, such as a prostate tumor, a pancreatic tumor, a gastric tumor, an ovarian tumor, etc., can be increased, decreased, or maintained. In certain embodiments, clinicians can make such a decision based on whether the presence, absence, or amount of the biomarker is below, above, or about the biomarker threshold, respectively. When clinical applications are contemplated, it is necessary to prepare the pharmaceutical compositions - expression constructs, expression vectors, fusion proteins, transduced cells, activated T cells, transduced and loaded T cells - in a form appropriate for the intended application. Combination therapy

[0335] To increase the effectiveness of the expression vectors presented herein, it may be desirable to combine these compositions and methods with agents effective in the treatment of disease.

[0336] In certain embodiments, anti-cancer agents can be used in combination with the method. An "anti-cancer" agent can adversely affect cancer in a subject by, for example, killing one or more cancer cells, inducing apoptosis of one or more cancer cells, reducing the growth rate of one or more cancer cells, reducing the incidence or number of metastases, reducing the size of a tumor, inhibiting the growth of a tumor, reducing the blood supply to a tumor or one or more cancer cells, promoting an immune response to one or more cancer cells or tumors, preventing or inhibiting the progression of cancer, or prolonging the lifespan of a subject with cancer. Anti-cancer agents include, for example, chemotherapeutic agents (chemotherapy), radiotherapy agents (radiotherapy), surgical procedures (surgery), immunotherapy agents (immunotherapy), gene therapy agents (gene therapy), hormone therapy, other biological agents (biological therapy) and / or alternative therapies.

[0337] In some embodiments, antibiotics can be used in combination with the pharmaceutical composition to treat and / or prevent infectious diseases, including amikacin, aminoglycosides (e.g., gentamicin), amoxicillin, amphotericin B, ampicillin, antimonials, atovaquone, sodium stibogluconate, azithromycin, capreomycin, cefotaxime, cefoxitin, ceftriaxone, chloramphenicol, clarithromycin, clindamycin, clofazimine, cycloserine, dapsone, doxycycline, ethambutol, ethionamide, fluconazole, fluoroquinolones, isoniazid, itraconazole, kanamycin, ketoconazole, minocycline, ofloxacin, para-aminosalicylic acid, pentamidine, polymyxin defensin, prothionamide, pyrazinamide, pyrimethamine. These include, but are not limited to, sulfadiazine, quinolones (eg, ciprofloxacin), rifabutin, rifampin, sparfloxacin, streptomycin, sulfonamides, tetracyclines, thiacetazone, trimethaprim-sulfamethoxazole, viomycin, or combinations thereof.

[0338] More generally, such agents are provided in a combined amount with an expression vector effective to kill or inhibit the growth of cancer cells and / or microorganisms. This process may include contacting a cell(s) with an agent(s) and a pharmaceutical composition, either simultaneously or within a period during which separate administration of the pharmaceutical composition and agent to the cell, tissue, or organism produces the desired therapeutic benefit. This may be accomplished by contacting the cell, tissue, or organism with a single composition or pharmacological formulation that contains both the pharmaceutical composition and one or more agents, or by contacting the cell with two or more separate compositions or formulations, one composition containing the pharmaceutical composition and the other containing one or more agents.

[0339] The terms "contacted" and "exposed" as applied to a cell, tissue, or organism are used herein to describe the process by which a pharmaceutical composition and / or another agent, such as a chemotherapeutic or radiotherapeutic agent, is delivered to or directly juxtaposed with a target cell, tissue, or organism. To effect cell killing or stasis, the pharmaceutical composition and / or additional agent are delivered to one or more cells in a combined amount effective to kill the cell or prevent the cell from dividing. In some embodiments, the chemotherapeutic agent is selected from the group consisting of carboplatin, estramustine phosphate (Emcyt), and thalidomide. In some embodiments, the chemotherapeutic agent is a taxane. The taxane may be selected from the group consisting of, for example, docetaxel (taxotere), paclitaxel, and cabazitaxel. In some embodiments, the taxane is docetaxel. In some embodiments, the chemotherapeutic agent is administered simultaneously with or within one week after administration of the modified cell or nucleic acid. In other embodiments, the chemotherapeutic agent is administered 1-4 weeks, or 1 week-1 month, 1 week-2 months, 1 week-3 months, 1 week-6 months, 1 week-9 months, or 1 week-12 months after administration of the modified cells or nucleic acid. In some embodiments, the chemotherapeutic agent is administered at least 1 month prior to administration of the cells or nucleic acid.

[0340] The administration of the pharmaceutical composition may precede, be combined with, or be subsequent to the other agent(s) by intervals ranging from minutes to weeks. In embodiments in which the pharmaceutical composition and the other agent(s) are applied separately to the cell, tissue, or organism, generally no significant period of time will elapse between the respective delivery times, so that the pharmaceutical composition and the agent(s) can still exert their beneficial combined effect on the cell, tissue, or organism. For example, in such cases, it is contemplated that the cell, tissue, or organism may be contacted with the pharmaceutical composition using two, three, four, or more modalities substantially simultaneously (i.e., within less than about one minute). In other aspects, the one or more agents may be administered substantially simultaneously, before and / or after administration of the expression vector, within a range of about one minute to about 24 hours, to about seven days, to about one to about eight weeks, or more, and any range derivable therein. Still further, various combination regimens of the pharmaceutical composition and one or more agents presented herein may be utilized.

[0341] In some embodiments, the chemotherapeutic agent may be a lymphodepleting chemotherapeutic agent.

[0342] In other examples, the chemotherapeutic agent can be taxotere (docetaxel) or another taxane, such as cabazitaxel. The chemotherapeutic agent can be administered before, during, or after treatment with cells and inducer. For example, the chemotherapeutic agent can be administered about 1 year, 11, 10, 9, 8, 7, 6, 5, or 4 months, or 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 weeks, or 1 week before administration of the first dose of activating nucleic acid. Or, for example, the chemotherapeutic agent can be administered about 1 week, or 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 weeks, or ... months, or 1 year after administration of the first dose of cells or inducer.

[0343] The administration of a chemotherapeutic agent may include administration of more than one chemotherapeutic agent, for example, cisplatin may be administered in addition to taxotere or other taxanes, such as cabazitaxel.

[0344] In some embodiments, the invention provides combination therapies comprising a modified cell population as described herein and a cytokine or chemokine neutralizing agent, e.g., a neutralizing antibody. In some embodiments, the invention provides combination therapies comprising a modified cell population as described herein and a TNFα neutralizing agent, e.g., an anti-TNFα antibody. Indications for adjusting or maintaining subsequent drug doses

[0345] An indication for adjusting or maintaining a subsequent drug dose may be based on the presence or absence of the biomarker. For example, if (i) a low sensitivity determination of the biomarker level was available, (ii) the biomarker level shifted sharply in response to the drug, (iii) low or high levels of the biomarker were present, and / or (iv) the drug is not appreciably toxic at the administered level, the presence or absence of the biomarker may be sufficient to generate an indication for adjusting or maintaining a subsequent drug dose.

[0346] The index for adjusting or maintaining the subsequent drug dose is often based on the amount or level of a biomarker (e.g., PSCA). The amount of a biomarker can be a mean value, a median value, a nominal value, a range, an interval, a maximum value, a minimum value, or a relative amount in some embodiments. The amount of a biomarker can be expressed with or without a measurement error range in certain embodiments.

[0347] The amount of biomarker can be expressed in some embodiments as biomarker concentration, weight of biomarker per unit weight, weight of biomarker per unit volume, moles of biomarker, moles of biomarker per unit volume, moles of biomarker per unit weight, weight of biomarker per unit cell, volume of biomarker per unit cell, moles of biomarker per unit cell, etc. Weight can be expressed, for example, in femtograms, picograms, nanograms, micrograms, milligrams, and grams. Volume can be expressed, for example, as femtoliters, picoliters, nanoliters, microliters, milliliters, and liters. Moles can be expressed, for example, in picomoles, nanomoles, micromoles, millimoles, and moles. In some embodiments, the unit weight can be the body weight of the subject or the weight of a sample from the subject, the unit volume can be the volume of a sample from the subject (e.g., blood volume), and the unit cell can be per cell or per a certain number of cells (e.g., micrograms of biomarker per 1000 cells). In some embodiments, the amount of a biomarker determined from one tissue or fluid can be correlated with the amount of the biomarker in another fluid or tissue known in the art.

[0348] An indication for adjusting or maintaining a subsequent drug dose is often generated by comparing the determined level of the biomarker in the subject with a predetermined level of the biomarker. The predetermined level of the biomarker may in certain embodiments be associated with a therapeutic or effective amount of the drug in the subject, may be associated with a toxic level of the drug, may be associated with the presence of a condition, may be associated with a midpoint of treatment, and may be associated with an end point of treatment. The predetermined level of the biomarker may include time as a factor, and in some embodiments, the threshold is a time-dependent signature.

[0349] Some treatment methods include the steps of (i) administering a drug to a subject in one or more doses (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 doses), (ii) determining the presence, absence or amount of a biomarker (e.g., PSCA) in or from the subject after (i), (iii) obtaining an indication to increase, decrease or maintain a subsequent dose of the drug for administration to the subject, and (iv) optionally administering a subsequent dose to the subject, where the subsequent dose is increased, decreased or maintained relative to the preceding dose in (i). In some embodiments, the presence, absence or amount of the biomarker is determined after each dose of the drug is administered to the subject, and the presence, absence or amount of the biomarker may not be determined after each dose of the drug is administered to the subject (e.g., the biomarker is evaluated after one or more of the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth or tenth doses, but not after each dose is administered).

[0350] The indication for adjusting the subsequent drug dose can also be considered as the need to increase or decrease the subsequent drug dose. The indication for adjusting or maintaining the subsequent drug regimen can be taken into consideration by the clinician, who may act on the indication in certain embodiments. In some embodiments, the clinician may not choose to act on the indication. Thus, the clinician can choose to adjust or not adjust the subsequent drug dose based on the indication provided.

[0351] The indicator for adjusting or maintaining the subsequent drug dose and / or the subsequent drug administration can be provided in a conventional manner. In some embodiments, the indicator can be provided in a table format (e.g., in a physical or electronic medium). For example, a biomarker threshold value may be provided in a table, and the clinician can compare the presence, absence, or amount of the biomarker determined for the subject with the threshold value. The clinician can then identify the indicator for the subsequent drug administration from the table. In certain embodiments, the indicator can be presented (e.g., displayed) by the computer after the presence, absence, or amount of the biomarker is provided to the computer (e.g., entered into the computer's memory). For example, the presence, absence, or amount of the biomarker determined for the subject can be provided to the computer (e.g., entered into the computer by a user or sent to the computer via a remote device in a computer network), and the computer's software can generate an indicator for adjusting or maintaining the subsequent drug administration and / or provide the subsequent drug administration. For example, subsequent doses can be determined based on certain factors other than the presence, absence or amount of a biomarker, such as the subject's body weight, the subject's level of one or more metabolites (e.g., metabolite levels associated with liver function), etc.

[0352] Once the subsequent dose is determined based on the indication, the clinician can administer the subsequent dose or can instruct another person or entity to adjust the dose. The term "clinician" as used herein refers to a decision maker, which in certain embodiments is a medical professional. The decision maker can in some embodiments be a computer or a displayed computer program output, and the medical service provider can act according to the indication or subsequent drug dose displayed by the computer. The decision maker can administer the subsequent dose directly (e.g., inject the subsequent dose into the subject) or can administer remotely (e.g., pump parameters can be changed remotely by the decision maker).

[0353] Subjects can be pre-screened to determine whether it is possible to determine the presence, absence or amount of a particular biomarker (e.g., PSCA).Non-limiting examples of pre-screening include: identifying the presence or absence of a genetic marker (e.g., polymorphism, particular nucleotide sequence); identifying the presence, absence or amount of a particular metabolite.Clinicians can use the pre-screening results in combination with the presence, absence or amount of a biomarker to determine whether the subsequent drug dosage can be adjusted or maintained. EXAMPLES

[0354] The examples provided below are illustrative of certain embodiments and are not intended to limit the present technology. Example 1 PSCA-specific CAR-T cell clinical research

[0355] The PSCA-specific CAR-T cells of this example are PSCA-directed, genetically engineered, autologous candidates that bind to PSCA-expressing cells. The PSCA-specific CAR-T cells target PSCA with a first-generation CAR construct that contains the traditional CD3ζ cytoplasmic signaling domain together with the single-chain variable region domain (scFV) of a humanized PSCA antibody. In addition to the anti-PSCA CAR, the PSCA-specific CAR-T cells were engineered to express an inducible dual costimulatory domain composed of two FK-binding proteins (FKBPs) in frame with the signaling domains from MyD88 and CD40 (inducible MyD88 / CD40 or iMC).

[0356] iMCs function as molecular switches to enhance and control T cell activation and proliferation in the presence of the small molecule dimerizer rimiduside (Figure 3). The tandem FKBP domains provide a ligand dimerization scaffold that induces iMCs in a rimiduside-dependent manner to initiate downstream signaling pathways that not only promote cell proliferation and survival but also upregulate inflammatory cytokines and type I interferons.

[0357] The inducible MyD88 / CD40 PSCA CAR retroviral construct contains, in the 5' to 3' direction, a polynucleotide encoding:

[0358] (1) a fusion protein (the entire fusion protein may be designated MC) containing a truncated human MyD88 polypeptide (the amino terminal 172 amino acids of MyD88 containing the DD and middle domains; SEQ ID NO:21, encoded by SEQ ID NO:22) fused to a portion of a human CD40 polypeptide (the carboxy terminal 62 amino acids, i.e., amino acids 216-277 of CD40; SEQ ID NO:23, encoded by SEQ ID NO:24) fused to two copies of a mutant human FKBP12 protein in which the phenylalanine at amino acid position 36 (or 37 counting the first methionine of the protein) has been replaced with valine (FKBP12(F36V), also known as FKBP12v36, FV36, FKBPv or Fv; SEQ ID NOs:41 and 43, encoded by SEQ ID NOs:42 and 44);

[0359] (2) T2A polypeptide (SEQ ID NO:5 encoded by SEQ ID NO:6); and

[0360] (3) A first generation CAR polypeptide comprising, in the 5' to 3' direction, an scFv sequence (including a membrane signal peptide (SEQ ID NO:7 encoded by SEQ ID NO:8) fused to the light and heavy chain variable regions of an anti-PSCA monoclonal antibody with a flexible 8 amino acid glycine-serine linker between the chains, i.e., a flex peptide (SEQ ID NO:9 encoded by SEQ ID NO:10)), a human CD34 epitope polypeptide to which it is fused (amino acids 30-45 of CD34; SEQ ID NO:11 encoded by SEQ ID NO:12), an alpha stalk of human CD8 to which it is fused (amino acids 141-182 of CD8; SEQ ID NO:13 encoded by SEQ ID NO:14), a transmembrane domain of human CD8 to which it is fused (amino acids 183-219 of CD8; SEQ ID NO:15 encoded by SEQ ID NO:16), and a portion of human CD3ζ to which it is fused (amino acids 83-194 of CD3ζ isoform X2; SEQ ID NO:17 encoded by SEQ ID NO:18).

[0361] Inducible MyD88 / CD40 PSCA CAR-T cells were rationally designed to proliferate in response to two distinct signals: 1) antigen-specific activation by PSCA recognition and CD3ζ signaling domain (signal 1), and 2) rimiduside-dependent costimulation by iMC (signal 2) (Figure 4). This design allows inducible MyD88 / CD40 PSCA CAR-T cells to be optimized for both antigen-directed and antigen-independent T cell activation, proliferation, and persistence, thereby resulting in enhanced clinical activity for the treatment of solid tumor malignancies compared to older CARs (signal 3; Figure 4).

[0362] To determine whether a dose-response relationship exists for antigen-dependent cytotoxicity and to assess whether iMCs may affect this, inducible MyD88 / CD40 PSCA CAR-T cells (0.625 × 10 6 cells, 1.25×10 6 cells, 2.5 x 10 6 cells, 5.0×10 6cells, and 10.0 × 10 6 Tumor-bearing NSG mice were treated with a single ascending dose of rimiduside (T cells) administered by intravenous injection on day 0 (Figure 5A, Figure 5B). When tumor measurements showed growth based on two consecutive measurements after T cell injection, weekly intraperitoneal rimiduside 5 mg / kg was initiated. By day 14, doses ≥ 2.5 × 10 6 The cells led to rapid control and eventual disappearance of HPAC tumors. 6 The cells were insufficient to reduce tumor volume in the absence of rimiduside. However, after iMC activation, tumor growth was reduced by 0.625 × 10 6 or 1.25×10 6 In mice receiving 1.0 × 10 iMCs, the IL-16 expression level was stabilized or reduced, respectively. Collectively, these data not only support that activation of iMCs with rimiduside in HPAC tumor-bearing mice can support antitumor function, but also demonstrate that activation of iMCs with rimiduside can support antitumor function in HPAC tumor-bearing mice. 6 The dose level is defined as the minimal dose level at which cells affect tumor growth in vivo. Furthermore, all inducible MyD88 / CD40 PSCA CAR-T cell-treated mice maintained or gained weight over the course of the experiment, regardless of T cell dose level with or without rimiducide treatment, suggesting that the treatment was well tolerated without any overt signs of toxicity.

[0363] The study depicted in Figure 7 was designed to determine the feasibility, safety, and clinical activity of PSCA-specific CAR-T cells administered with rimiduside in subjects with previously treated PSCA-positive advanced solid tumors (pancreatic, gastric / GEJ, prostate cancer).

[0364] FIG. 6 shows PSCA screening of pancreatic tumors.

[0365] Part 1 (Phase 1): Inducible MyD88 / CD40 PSCA CAR-T cells (1.25 × 10 6 cells / kg~5.0×10 6Cell dose escalation to identify the maximum dose (escalating dose of cells / kg).

[0366] Part 2 (Phase 2): Indication-specific dose expansion to evaluate safety, pharmacodynamics (including inducible MyD88 / CD40 PSCA CAR-T cell persistence), and clinical activity at the doses identified in Part 1 in a variety of PSCA-expressing solid tumors.

[0367] The initial starting dose of inducible MyD88 / CD40 PSCA CAR-T cells was 1.25 × 10 6 Rimiducide was administered at a fixed dose of 0.4 mg / kg.

[0368] Cohort 0 was performed as follows: 3 subjects were enrolled and administered PSCA CAR-T cells (1.25 × 10 6 These subjects were administered a fixed, single dose of rimiducide (0.4 mg / kg) at 4 °C for 12 h. No dose-limiting toxicity events were observed, and there were no adverse events (AEs) elevating cytokine levels or other signs of toxicity associated with PSCA CAR-T cell infusion. Quantitative polymerase chain reaction (qPCR) analysis of peripheral blood samples revealed that PSCA CAR-T cell numbers increased approximately between days 0 and 15, with maximal expansion observed between days 5 and 15, before eventually declining below assay detection levels by day 21. Based on the favorable safety and early clearance of PSCA CAR-T cells demonstrated in cohort 0, a fixed, single dose of rimiducide (0.4 mg / kg) was added to the investigational treatment regimen on day 7.

[0369] Cohort 3 (1.25 × 10 6 The first cohort in part 1 was the first to begin cell dose escalation determination for PSCA CAR-T cells administered with rimducaside (0.4 mg / kg cells). A cohort of 3 subjects was treated with escalating doses of PSCA CAR-T cells on day 0 and a fixed, single dose of rimducaside (0.4 mg / kg) on ​​day 7. The doses of CAR-T cells are listed in Figure 8.

[0370] Results are presented in Figures 9-14. A safety summary is provided in Figure 9. No DLTs, neurotoxicity, or cytokine release syndrome events were observed. Fever was the only treatment-related AE reported by two or more patients. Both events resolved within 24-36 hours with supportive care. Specifically, one patient in cohort 4 experienced grade 1 fever after infusion of inducible MyD88 / CD40 PSCA CAR-T cells on day 0, an event that resolved within 24 hours with paracetamol. One patient in cohort 5 experienced grade 2 fever after infusion of inducible MyD88 / CD40 PSCA CAR-T cells on day 0. This event resolved within 36 hours with paracetamol, but was reported as an SAE because the fever prolonged the patient's planned infusion-related hospitalization. No other treatment-related SAEs were reported. Both fever events were associated with time-matched, transient peripheral cytokine increases. Grade 1 events were associated with moderate hypotension that resolved within 2 hours with IV fluids, and grade 2 events were isolated and occurred in the absence of other systemic clinical symptoms. Six patients reported 14 SAEs, and all but one SAE (grade 2 fever) was related to the disease under study.

[0371] Expansion and persistence of inducible MyD88 / CD40 PSCA CAR-T cells is shown in Figure 10. Only slight evidence of lymphodepletion (LD) (79% ± 25% of cells remained) was observed with the cyclophosphamide (CTX)-only regimen. Rapid cell expansion was observed by day 4, but was not sustained without rimiduside. Rapid cell engraftment was observed by day 4 in the majority of patients, despite insufficient lymphodepletion in all but one subject. The mean percentage of lymphocytes remaining after lymphodepletion with cyclophosphamide was 79% (SD 25%). Of the eight patients who received a single dose of rimiduside, four had at least 3-fold and up to 20-fold cell expansion within 7 days, and three had cell persistence for more than 3 weeks after rimiduside infusion. Cell persistence for more than 4 months was observed in one patient, who also had the highest PSCA expression at screening. In cohort 0, only modest peripheral expansion was observed when inducible MyD88 / CD40 PSCA CAR-T cells were infused alone.

[0372] Peripheral cytokine profiles over time are shown in Figure 11. All eight patients who received rimduca had increases in cytokines (IP-10, TNF-α) that correlated with cell expansion. Increases in serum IP-10 indicate prior IFNγ production, and along with increases in serum TNF-α, support T cell activation in response to activation of iMCs with rimduca. Increases in cell dose are associated with increased overall serum cytokine and chemokine levels. Increased levels of chemokines, including MCP-1, MIP-1α and MIP-1β, which are produced by activated lymphocytes and macrophages and have important chemotactic and proinflammatory functions, were observed in many patients after rim infusion. Reductions in peripheral lymphocytes (associated with increased serum IL-7 and IL-15 levels) were observed in one patient, and increases in IL-7 and IL-15 after lymphodepletion were associated with reductions in peripheral lymphocyte counts to 31% of starting levels. Overall, lymphocytes were reduced to an average of 78.7±25% of starting levels.

[0373] Evidence of anti-tumor activity in patients treated with inducible MyD88 / CD40 PSCA CAR-T cells is shown in Figure 12 and Figure 13. Scans were performed at week 4 (month 1), week 8 (month 2), and every 8 weeks thereafter (every 2 months) until progression. Two patients had tumor shrinkage of more than 20%.

[0374] Figure 14 shows the time to next treatment following administration of PSCA CAR-T cells.

[0375] Thus, taken together, these results indicate that administration of PSCA CAR-T cells and a single dose of rimiduside was well tolerated. No cytokine release syndrome or neurotoxicity of any grade was observed. The most frequent AEs were consistent with those experienced by patients with advanced cancer receiving cytotoxic chemotherapy or other cancer immunotherapies.

[0376] Treatment-related AEs were few and mild to moderate in intensity and resolved with supportive care. In addition, PSCA CAR-T cells demonstrated enhanced expansion and prolonged persistence in some patients, despite the lack of lymphodepletion with cyclophosphamide alone. Thus, bioactivity / disease control was observed in this heavily pretreated patient population. method

[0377] Description of the manufacturing process: PSCA CAR-T cells are prepared from patients' peripheral blood mononuclear cells (PBMCs), which are obtained by standard apheresis procedures as described below. PBMCs are enriched for T cells, which are then washed and expanded in cell culture. Once the target number of cells is available, the expanded T cells are transduced with a replication-incompetent retroviral vector containing the anti-PSCA CAR and the rimiduside-inducible iMC transgene. The transduced T cells are formulated into a suspension and cryopreserved. The final PSCA CAR-T cell product must pass release testing, including sterility testing, before being shipped as a frozen suspension in patient-specific infusion bags.

[0378] Packaging and formulation: PSCA CAR-T cells are cryopreserved in 10-15 mL of freezing medium and stored frozen in cryopreservation bags in the vapor phase of liquid nitrogen. Apheresis

[0379] Peripheral blood mononuclear cells (PBMCs) will be collected from the subject according to the site's standards. Prior to apheresis, the subject's white blood cell count and differential cell counts will be measured and recorded. Results should demonstrate that the subject has adequate bone marrow function that is not transfusion dependent and not supported by growth factors: Absolute lymphocyte count ≥ 1 × 10 2 White blood cell count ≥ 2 x 10 / µL 3 / μL Absolute neutrophil count ≥ 1 × 10 without granulocyte colony-stimulating factor support 3 / μL Platelets ≥ 100 × 10 3 / μL Hemoglobin ≥ 9g / dL or as per apheresis institution guidelines.

[0380] One to two blood volumes were drawn and processed using standardized continuous-flow centrifugation according to the institution's standard procedures. The volume processed and the duration of apheresis were recorded. Subjects were monitored for infectious diseases during and after the apheresis procedure per established regulatory guidelines. Before pre-apheresis evaluation, venous access, monitoring and treatment of apheresis complications were performed according to institution guidelines. Lymphocyte depletion prior to PSCA CAR-T cell infusion

[0381] Cyclophosphamide 500 mg / m IV 2 and fludarabine 30 mg / m intravenously. 2was administered 5, 4, and 3 days prior to infusion of PSCA CAR T cells unless the combination was not tolerated. If it was determined that combination-based lymphodepletion posed an unfavorable safety risk to the subject, lymphodepletion was pursued with cyclophosphamide alone. Example 2 Nucleic acid and amino acid sequences

[0382] Table 1: Amino acid sequences [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0383] Table 2: Nucleic acid sequences [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] Example 3 Example of an embodiment

[0384] The examples provided below are illustrative of certain embodiments and are not intended to limit the present technology.

[0385] A1. A heterogeneous T cell population comprising modified and unmodified T cells, wherein the modified T cells comprise a) a first polynucleotide encoding an inducible MyD88 / CD40 polypeptide, and b) a second polynucleotide encoding a prostate stem cell antigen-chimeric antigen receptor (PSCA-CAR) po...

Claims

[Claim 1] The invention described in the specification.

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