scholarly journals Preclinical Evaluation of Prgn-3007, a Non-Viral, Multigenic, Autologous ROR1 Ultracar-T ® Therapy with Novel Mechanism of Intrinsic PD-1 Blockade for Treatment of Hematological and Solid Cancers

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 1694-1694
Author(s):  
Tim Chan ◽  
Sean P Scott ◽  
Mengyan Du ◽  
Cheryl Bolinger ◽  
Carol Poortman ◽  
...  
Keyword(s):  
T Cells ◽  
Manufacturing Process ◽  
Cell Lymphoma ◽  
Viral Gene ◽  
Publicly Traded ◽  
Car T Cells ◽  
Tumor Bearing ◽  
Car T ◽  
Mantle Cell ◽  
Viral Gene Delivery

Abstract Traditional methods for chimeric antigen receptor (CAR) T manufacturing utilize viral vectors, ex vivo activation and expansion of T cells to achieve clinically relevant cell numbers, which leads to an exhausted T cell phenotype, high manufacturing costs, and treatment delays. The UltraCAR-T platform is designed to overcome these limitations using our advanced non-viral gene delivery system and a rapid, overnight manufacturing process (Blood 2019 134 (Supplement_1):2660; Blood 2020 136 (Supplement 1):17); Cancer Research 2020 80 (16Suppl):6593). UltraCAR-T cells, which express antigen specific CAR, membrane-bound IL-15 (mbIL15), and kill switch genes, are manufactured at the medical center's cGMP facility using autologous T cells and administered back to the patient only one day after gene transfer. UltraCAR-T cells are currently under clinical investigation for hematological (NCT03927261) and solid tumors (NCT03907527). Here we describe the advancement of the UltraCAR-T platform to address the inhibitory tumor microenvironment by incorporating intrinsic checkpoint blockade without the need for complex and expensive gene editing techniques. PRGN-3007, based on the next generation of the UltraCAR-T platform, is engineered to simultaneously express CAR for targeting receptor tyrosine kinase-like orphan receptor 1 (ROR1), which is overexpressed on many hematological and solid tumors; mbIL15 for enhanced in vivo expansion and persistence; kill switch for improved safety profile; and a novel mechanism for the intrinsic blockade of PD-1 gene expression. This approach of intrinsic blockade of PD-1 expression, only on UltraCAR-T cells, is aimed to avoid systemic toxicity and high cost of checkpoint inhibitors by eliminating the need for combination treatment. PRGN-3007 is manufactured using the already established rapid and streamlined UltraCAR-T manufacturing process. PRGN-3007 was generated using multiple healthy donor T cells using multi-cistronic non-viral vector and the overnight manufacturing process. The co-expression of CAR, mbIL15 and kill switch transgenes was confirmed by flow cytometry, western blotting, and qPCR. Furthermore, PRGN-3007 showed significant reduction in PD-1 expression on UltraCAR-T cells compared to ROR1 CAR-T cells lacking PD-1 blockade (Control ROR1 CAR-T). The downregulation of PD-1 expression on PRGN-3007 resulted in enhanced ROR1-specific cytotoxicity and release of inflammatory cytokines upon co-culture with various ROR1 + PD-L1 + hematological and solid tumor cells compared to Control ROR1 CAR-T, especially at low effector to target cell ratios. Single-cell cytokine proteomics showed that the downregulation of PD-1 expression on PRGN-3007 resulted in a significantly higher number of polyfunctional CAR-T cells compared to Control ROR1 CAR-T. Expression of mbIL15 on UltraCAR-T, with or without downregulation of PD-1 expression, resulted in robust expansion in presence of ROR1 antigen, lack of autonomous expansion in absence of ROR1, and durable persistence even in absence of exogenous cytokines in vitro. Furthermore, PRGN-3007 was selectively and effectively eliminated by the kill switch activator antibody treatment. A single administration of PRGN-3007, only one day after gene transfer, effectively reduced tumor burden and significantly improved overall survival (p<0.05) of tumor bearing mice compared to Control ROR1 CAR-T in an aggressive xenograft model of mantle cell lymphoma (Figure). Blood analyses demonstrated sustained downregulation of PD-1 expression, rapid expansion, long-term persistence, and a predominant central memory phenotype of PRGN-3007 in tumor bearing mice. In summary, these preclinical data highlight the overall safety and improved efficacy of incorporating intrinsic downregulation of PD-1 expression on UltraCAR-T cells using non-viral gene delivery and the established rapid, decentralized manufacturing process. These data provide a strong rationale for the evaluation of PRGN-3007 for the treatment of ROR1 + malignancies. Figure: Overall survival in in an established model of mantle cell lymphoma in NSG mice. Tumor cells were engrafted in mice on Day 0 and treatments were administered on Day 8. Data shown is from 8 mice/group at the start of the study. * p<0.05, ***p<0.001; log rank test. Figure 1 Figure 1. Disclosures Chan: Precigen, Inc: Current Employment, Current equity holder in publicly-traded company. Scott: Precigen, Inc: Current Employment. Du: Precigen, Inc: Current Employment. Bolinger: Precigen, Inc: Current Employment. Poortman: Precigen, Inc: Current Employment. Shepard: Precigen, Inc: Current Employment. Koenitzer: Precigen, Inc: Current Employment. Govekung: Precigen, Inc: Current Employment. Sailor: Precigen, Inc: Current Employment. Johnson: Precigen, Inc: Current Employment. Plummer: Precigen, Inc: Current Employment. Zilko: Precigen, Inc: Current Employment. Dutta: Precigen, Inc: Current Employment. Kunchithapautham: Precigen, Inc: Current Employment. Athwal: Precigen, Inc: Current Employment. Klocke: Precigen, Inc: Current Employment. Zinser: Precigen, Inc: Current Employment. Abdeladhim: Precigen, Inc: Current Employment. Ahmad: Precigen, Inc: Current Employment; Kite, A Gilead Company: Ended employment in the past 24 months. Metenou: Precigen, Inc: Current Employment. Semnani: Precigen, Inc: Current Employment. Brough: Precigen, Inc: Current Employment, Current equity holder in publicly-traded company. Shah: Precigen: Current Employment, Current equity holder in publicly-traded company. Sabzevari: Precigen: Current Employment, Current equity holder in publicly-traded company; Kinnate BioPharma: Membership on an entity's Board of Directors or advisory committees; Compass Therapeutics: Current equity holder in publicly-traded company.

scholarly journals A BAFF ligand-based CAR-T cell targeting three receptors and multiple B cell cancers

2022 ◽  
Vol 13 (1) ◽  
Author(s):  
Derek P. Wong ◽  
Nand K. Roy ◽  
Keman Zhang ◽  
Anusha Anukanth ◽  
Abhishek Asthana ◽  
...  
Keyword(s):  
T Cells ◽  
B Cell ◽  
Lymphoblastic Leukemia ◽  
Viral Gene ◽  
Car T Cells ◽  
Car T ◽  
Almost All ◽  
Viral Gene Delivery

AbstractB cell-activating factor (BAFF) binds the three receptors BAFF-R, BCMA, and TACI, predominantly expressed on mature B cells. Almost all B cell cancers are reported to express at least one of these receptors. Here we develop a BAFF ligand-based chimeric antigen receptor (CAR) and generate BAFF CAR-T cells using a non-viral gene delivery method. We show that BAFF CAR-T cells bind specifically to each of the three BAFF receptors and are effective at killing multiple B cell cancers, including mantle cell lymphoma (MCL), multiple myeloma (MM), and acute lymphoblastic leukemia (ALL), in vitro and in vivo using different xenograft models. Co-culture of BAFF CAR-T cells with these tumor cells results in induction of activation marker CD69, degranulation marker CD107a, and multiple proinflammatory cytokines. In summary, we report a ligand-based BAFF CAR-T capable of binding three different receptors, minimizing the potential for antigen escape in the treatment of B cell cancers.

scholarly journals CAR T Cells for Mantle Cell Lymphoma: Is it Time to Reshuffle the Deck?

Cancer Cell ◽  
2020 ◽  
Vol 37 (6) ◽  
pp. 761-763
Author(s):  
Craig S. Sauter ◽  
Renier J. Brentjens
Keyword(s):  
T Cells ◽  
Mantle Cell Lymphoma ◽  
Cell Lymphoma ◽  
Car T Cells ◽  
Car T ◽  
Mantle Cell

scholarly journals Phase 1 Study of CD37-Directed CAR T Cells in Patients with Relapsed or Refractory CD37+ Hematologic Malignancies

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 653-653
Author(s):  
Matthew J. Frigault ◽  
Yi-Bin Chen ◽  
Kathleen M.E. Gallagher ◽  
Nora K. Horick ◽  
Areej El-Jawahri ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Stock Options ◽  
Research Funding ◽  
Ad Hoc ◽  
Cell Lymphoma ◽  
Publicly Traded ◽  
Car T Cells ◽  
Advisory Boards ◽  
Car T

Abstract Background: CD37 is a tetraspanin molecule expressed in B-cell and some T-cell lymphomas. We designed a Chimeric Antigen Receptor (CAR) targeting CD37 and with 4-1BB and CD3z intracellular signaling domains (CART37). Preclinical studies indicated comparable anti-tumor activity to CD19-directed CAR-T cells against B-cell lymphomas, promising activity against CD37 + T-cell lymphomas, and no evidence of off-tumor activity (PMID: 30089630). The lentiviral vector used in the clinical study includes a truncated EGFR reporter gene. NCT04136275 is a Phase 1, single-site, open-label, dose-escalation trial enrolling subjects with lymphoma who have received ≥ 2 prior regimens, and whose tumor cells express CD37. Methods: We developed a clinical assay to assess CD37 expression on patient tumor samples using flow cytometry and IHC. Peripheral blood mononuclear cells are collected via leukapheresis and manufactured using the CliniMACS Prodigy®. Following release testing, fresh or cryopreserved cell product is infused. Subjects undergo lymphodepletion with fludarabine and cyclophosphamide, then receive CART37 as a single infusion. Planned starting dose was 100 x 10 6 CAR + T cells, with options to dose-escalate to 300 x 10 6 CAR + T cells or dose de-escalate to 30 x 10 6 CAR + T cells in the event of dose-limiting toxicities (DLT) using a 3+3 design. The primary outcome measure is incidence of adverse events (AEs), including DLTs. Additional outcome measures are clinical response, progression-free and overall survival; correlative studies focus on quantification and persistence of CAR + cells in blood, residual tumor, and cytokine modulation. Results: As of July 13, 2021, 4 subjects (ages 35-70 years) have received CART37. Subjects had a median of 5.5 prior lines of systemic therapy. Two subjects had primary refractory double-hit high-grade B cell lymphoma (HGBCL) that had relapsed after commercial CD19 CAR-T; one subject had cutaneous T cell lymphoma relapsed after extracorporeal photopheresis, alemtuzumab, total skin electron beam radiation, allogeneic hematopoietic stem cell transplant (HSCT), brentuximab, and donor lymphocyte infusion, and one subject had Hodgkin's lymphoma refractory to six prior regimens, including chemotherapy, brentuximab vedotin, nivolumab and everolimus. All subjects were infused in the DL1 cohort, but one subject (with cutaneous T cell lymphoma) received only 19 x 10 6 CAR + due to limited ex vivo expansion. Three subjects developed low-grade CRS and ICANS, and one subject developed refractory Grade 3 CRS and Grade 3 ICANS which resolved with medical management. One patient with HGBCL developed CD19 neg and CD37 neg progressive disease prior to the day 28 evaluation. The three other subjects demonstrated deep responses (2 CR, 1 PR that converted to CR) as best response. Two subjects are alive 208 and 272 days from CAR37 infusion. All subjects had detectable expansion of CART37 by flow cytometry and molecular assays. Two subjects (who received ≥ 100 x 10 6 CART37 had robust expansion and developed prolonged pancytopenia with marrow aplasia; cetuximab infusion decreased detection of truncated EGFR on circulating T cells but had no impact on vector copy number. Both subjects underwent allogeneic HSCT after conditioning with flu/cy/TBI(400) and post-transplant cyclophosphamide (PTCy) based GVHD prevention and successfully engrafted, and had no detectable CART37 after HSCT. Conclusions: In this initial cohort, CART37 infusion resulted in CRS or ICANs as is common for CAR T cell products. Bone marrow aplasia was unexpected and was observed in two subjects who received at least 100 x 10 6 CART37; this was successfully rescued with allogeneic HSCT. Three of four subjects had deep clinical responses in heavily pretreated, refractory disease of diverse lymphoma subtypes. The protocol is open to enrollment with dose de-escalation to 30 x10 6 CART37 and has been amended to require identification of a potential donor prior to treatment in the case that rescue allogeneic HSCT is needed. CART37 has a potential role in enabling allogeneic transplantation in patients with relapsed or refractory hematologic malignancies. Disclosures Frigault: Takeda: Consultancy; Editas: Consultancy; BMS: Consultancy; Novartis: Consultancy, Research Funding; Iovance: Consultancy; Arcellx: Consultancy; Kite: Consultancy, Research Funding. Chen: Gamida: Consultancy; Incyte: Consultancy. Wehrli: CSL Behring: Patents & Royalties; Nestle: Current equity holder in publicly-traded company; Novartis: Current equity holder in publicly-traded company. Spitzer: Bluebird Bio: Consultancy; Jazz Pharmaceuticals: Consultancy; Qihan Bio: Consultancy; Syneos Health: Consultancy. Preffer: Cytek Biosciences: Other: Unspecified Relationship. Shaw: Orchard Therapeutics, Ltd: Current equity holder in publicly-traded company. Nikiforow: Kite/Gilead: Other: Ad hoc advisory boards; Novartis: Other: Ad hoc advisory boards; Iovance: Other: Ad hoc advisory boards; GlaxoSmithKline (GSK): Other: Ad hoc advisory boards. Ritz: Amgen: Research Funding; Equillium: Research Funding; Kite/Gilead: Research Funding; Avrobio: Membership on an entity's Board of Directors or advisory committees; Akron: Consultancy; Biotech: Consultancy; Blackstone Life Sciences Advisor: Consultancy; Clade Therapeutics, Garuda Therapeutics: Consultancy; Immunitas Therapeutic: Consultancy; LifeVault Bio: Consultancy; Novartis: Consultancy; Rheos Medicines: Consultancy; Talaris Therapeutics: Consultancy; TScan Therapeutics: Consultancy. Maus: Novartis: Consultancy; Micromedicine: Consultancy, Current holder of stock options in a privately-held company; Kite Pharma: Consultancy, Research Funding; GSK: Consultancy; Intellia: Consultancy; In8bio (SAB): Consultancy; CRISPR therapeutics: Consultancy; Cabaletta Bio (SAB): Consultancy; BMS: Consultancy; Bayer: Consultancy; Atara: Consultancy; AstraZeneca: Consultancy; Astellas: Consultancy; Arcellx: Consultancy; Agenus: Consultancy; Adaptimmune: Consultancy; WindMIL: Consultancy; Tmunity: Consultancy; Torque: Consultancy, Current holder of stock options in a privately-held company; tcr2: Consultancy, Divested equity in a private or publicly-traded company in the past 24 months; century: Current equity holder in publicly-traded company; ichnos biosciences: Consultancy, Current holder of stock options in a privately-held company.

scholarly journals Relapsed/Refractory Diffuse Large B-Cell Lymphoma and Mantle Cell Lymphoma: Is It Time to Consider CAR-T for All?

2020 ◽  
Vol 18 (12.5) ◽  
pp. 1764-1766
Author(s):  
Andrew D. Zelenetz
Keyword(s):  
T Cells ◽  
B Cell ◽  
Cell Lymphoma ◽  
B Cell Lymphoma ◽  
Car T Cells ◽  
Non Hodgkin Lymphoma ◽  
Large B Cell Lymphoma ◽  
Car T ◽  
Mantle Cell ◽  
Large B Cell

CAR T cells have demonstrated activity in relapsed/refractory non-Hodgkin lymphoma, including diffuse large B-cell lymphoma (DLBCL), indolent non-Hodgkin lymphoma, and mantle cell lymphoma. For patients with chemorefractory disease, CAR T cells can provide a durable complete response in a portion of patients, which represents a major advance in the field. For patients with chemosensitive disease, however, additional data are needed to determine whether CAR T cells are preferable to conventional approaches. Studies in DLBCL have shown that patients experiencing a PET-positive partial response after second-line chemotherapy have long-term outcomes after high-dose therapy and autologous stem cell rescue that are similar to CAR T-cell therapy, with decreased toxicity and cost. Alternative third-line options such as tafasitamab/lenalidomide and bispecific antibodies may also have a role for patients with chemorefractory disease.

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