Showing papers by "Helen Sabzevari published in 2023"
TL;DR: The role of IL-35 in systemic sclerosis (SSc) patients was investigated in this article , focusing on CD4+ T cell response and immunomodulatory cytokine production.
Abstract: This study investigated the role of IL-35 in systemic sclerosis (SSc) patients, focusing on CD4+ T cell response and immunomodulatory cytokine production. By comparing the cytokine levels in healthy donors (HD) and SSc patients using ELISAs, we found a significantly lower plasma IL-35 concentration in the SSc patients (52.1 ± 5.6 vs. 143 ± 11.1, p < 0.001). Notably, the IL-35 levels showed a negative correlation with TGF-β (p < 0.001) and IL-17 (p = 0.04). Assessing the IL-35R expression across cell types in the SSc patients and HDs via flow cytometry, we found higher levels on monocytes (40.7 + 5.7 vs. 20.3 ± 1.9, p < 0.001) and lower levels on CD8+ T cells (61.8 ± 9.2 vs. 83.4 ± 0.8, p < 0.05) in the SSc patients. The addition of recombinant IL-35 to stimulated peripheral blood mononuclear cells reduced the IL-17+CD4+ T cell percentage (9.0 ± 1.5 vs. 4.8 ± 0.7, p < 0.05) and increased the IL-35+CD4+ T percentage (4.1 ± 2.3 vs. 10.2 ± 0.8, p < 0.001). In a Treg:Tresponder cell Sco-culture assay with HD and SSc samples, rIL35 decreased the cell proliferation and levels of IL-17A (178.2 ± 30.5 pg/mL vs. 37.4 ± 6.4 pg/mL, p < 0.001) and TGF-β (4194 ± 777 pg/mL vs. 2413 ± 608 pg/mL, p < 0.01). Furthermore, we observed a positive correlation between the modified Rodnan skin score (mRSS) and TGF-β (p < 0.001), while there was a negative correlation between mRSS and IL-35 (p = 0.004). Interestingly, higher levels of plasmatic IL-35 were detected in individuals with limited disease compared to those with diffuse disease (60.1 ± 8.0 vs. 832.3 ± 4.1, p < 0.05). These findings suggest that IL-35 exhibits anti-inflammatory properties in SSc and it may serve as a marker for disease severity and a therapeutic target.
TL;DR: In this paper , Giorgio Zenere et al. developed a robust in vitro and in vivo model of continuous tumor antigen exposure to evaluate antitumor activity of the UltraCAR-T cells.
Abstract:
The UltraCAR-T® platform, with decentralized overnight manufacturing, has shifted the autologous CAR-T manufacturing paradigm. This disruptive approach has been validated in clinical and pre-clinical studies for both hematologic and solid tumors. UltraCAR-T cells, manufactured using an overnight process with minimal ex vivo manipulation of autologous T cells. UltraCAR-T cells are engineered to co-express CAR, membrane bound IL-15, and kill switch genes using non-viral gene transfer via high-throughput UltraPorator™ system. Multigenic design and overnight manufacturing renders UltraCAR-T cells with enhanced in vivo expansion and persistence for durable antitumor activity.
Chronic antigen stimulation from the tumor can lead to CAR T having an exhausted phenotype contributing to treatment failures. Next generation UltraCAR-T cells also incorporate a novel cell intrinsic blockade of PD-1, superseding the need for combination therapy with checkpoint inhibitors and mitigate classic T cell exhaustion that occurs from chronic stimulation, thereby expanding the therapeutic window for efficacy. Using mesothelin (MSLN) as an exemplar from the CAR-target library, we developed a robust in vitro and in vivo model of continuous tumor antigen exposure to evaluate antitumor activity of the UltraCAR-T cells.
MSLN UltraCAR-T cells were generated with and without intrinsic PD-1 blockade. In vitro, UltraCAR-T cells with intrinsic PD-1 blockade demonstrated enhanced inflammatory cytokine expression and cytotoxicity at low effector:target ratios. In long-term co-culture assays with recurring challenge of MSLN+ PD-L1+ tumor cells every 2-3 days, UltraCAR-T cells with intrinsic PD-1 blockade showed sustained potent cytotoxicity which was superior to CAR-T cells lacking PD-1 blockade and in combination with αPD-1 antibody. Intrinsic PD-1 blockade markedly enhanced polyfunctionality of UltraCAR-T cells in the presence of MSLN+ PD-L1+ tumor.
In xenograft solid tumor model, a single administration of MSLN UltraCAR-T cells manufactured using UltraPorator demonstrated significant antitumor efficacy. Blood analysis showed sustained downregulation of PD-1, robust antigen specific expansion and durable persistence with central memory/stem-cell memory as the dominant phenotype of UltraCAR-T in vivo. Rechallenging the mice, who became tumor-free after a single UltraCAR-T infusion, with tumor for a second time to simulate tumor relapse led to the significant reduction in tumor burden without additional UltraCAR-T treatment.
Collectively, these pre-clinical data highlight the improved efficacy of intrinsic PD-1 blockade in next generation UltraCAR-T cells using non-viral gene delivery and an established rapid, decentralized manufacturing process.
Citation Format: Giorgio Zenere, Carol Poortman, Afreen Sayed, Devi Gunasekera, Cheryl Bolinger, Pooja Chauhan, Jacques Plummer, Shamim Ahmad, Rutul R. Shah, Helen Sabzevari, Douglas E. Brough, Tim Chan. Next generation UltraCAR-T® cells with intrinsic checkpoint inhibition and overnight manufacturing overcome suppressive tumor microenvironment leading to sustained antitumor activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1791.
TL;DR: In this paper , the authors evaluated the safety and recommended Phase 2 dose (RP2D) of PRGN-3005 in pts with r/r ovarian cancer (NCT03907527).
Abstract: 5590 Background: Relapsed or refractory (r/r) ovarian cancer patients (pts) have limited treatment options. PRGN-3005 UltraCAR-T cells express a chimeric antigen receptor (CAR) targeting unshed MUC16, membrane bound IL-15 (mbIL15) and a kill switch with robust activity in preclinical models. UltraCAR-T investigational therapies are manufactured overnight using the proprietary UltraPorator electroporation system at the medical center and administered to patients one day after gene transfer. Methods: The Phase 1/1b study of PRGN-3005 evaluates the safety and recommended Phase 2 dose (RP2D) of PRGN-3005 in pts with r/r ovarian cancer (NCT03907527). In Phase 1, pts received PRGN-3005 without lymphodepletion (LD) via Intraperitoneal (IP) infusion (Cohort 1, C1) or Intravenous (IV) infusion (Cohort 2, C2) at Dose level 1-3. Dose level 3, IV infusion was further evaluated in pts treated with cyclophosphamide LD (60mg/kg/day on days -4 to -3 (IV LD)). As of the February 6, 2023 data cut-off, 25 evaluable pts have been treated in C1 (n = 12), C2 (n = 6), and IV LD (n = 7). Pts had a median age of 64 years (38-76) and were heavily pre-treated with a median of 8 prior regimens (4-11). Pts treated in C1 received 1 to 65.5 x 105 cells/kg, pts treated in C2 received 0.1 to 50 x 105 cells/kg and pts treated in IV LD received 0.1 to 39 x 105 cells/kg. Results: PRGN-3005 was well tolerated at up to 65.5 x 105 cells/kg. There have been no deaths, dose-limiting toxicities, neurotoxicity, grade ≥3 Cytokine Release Syndrome (CRS), or unexpected on-target/off-target toxicities related to PRGN-3005, and no use of the kill switch as of data cut-off. Grade 1 CRS occurred in 3/7 IV LD pts and one pt had transient grade 2 CRS, which resolved in < 24 hours. Plasma levels of IL-15 did not increase with treatment confirming mbIL15 is not shed. A dose dependent increase in PRGN-3005 was observed in peripheral blood of patients even in the absence of LD, and i n vivo persistence of PRGN-3005 could be observed in some patients for up to 9 months post infusion. Response was evaluated per RECIST 1.1, and 20% of all subjects experienced a response in at least 1 target lesion. In the IV LD pts, the disease control rate (DCR) was 85.7% at first restaging, with a decreased target tumor burden in 4/7 (57%), and a 27.4% average decrease in CA125. One pt had a 28% decrease in target tumor burden after receiving a second infusion of PRGN-3005 after 12 months. Conclusions: PRGN-3005 UltraCAR-T targeting MUC16 has been well tolerated with minimal toxicity. PRGN-3005 demonstrated a dose-dependent expansion and persistence in blood with or without LD. Encouraging DCR rates and a reduction in overall tumor burden have been observed in heavily pretreated, ovarian cancer pts treated with LD. The study continues to enroll to the Phase 1b expansion phase and is treating pts at DL3 with LD with an option to receive a second dose. Clinical trial information: NCT03907527 .