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149 Prolonged Persistence and Functional Activity of Anti-BCMA CAR-T Durcabtagene Autoleucel (PHE885) Manufactured Using the T-Charge™ Platform

Program: Oral and Poster Abstracts
Type: Oral
Session: 711. Cell Collection and Manufacturing of HSPCs, CAR-T Cells, and Other Cellular Therapy Products: Innovations in Mobilization, Collection, and Manufacturing for Cellular Therapies
Hematology Disease Topics & Pathways:
Adult, Research, Translational Research, Chimeric Antigen Receptor (CAR)-T Cell Therapies, Plasma Cell Disorders, Diseases, Biological therapies, Treatment Considerations, Lymphoid Malignancies, Human
Saturday, December 7, 2024: 1:00 PM

Shuntaro Ikegawa, MD, PhD1*, Adam S. Sperling, MD, PhD1,2, Sarah Nikiforow, MD, PhD1,2, Andrew Ghazi, PhD3*, Anthony Christidis, PhD3*, David Quinn, BA4*, Dexiu Bu, MD, PhD4*, Jennifer Mataraza, PhD4*, David Pearson, PhD5*, Allison Charles, BS4*, Marc Credi, MA4*, Nina Orwitz, PhD6*, Madisyn Strange1*, Rakuyo Tamada, BA1*, Kevin M. Panaro, BS1*, Casey Gervais, MS1*, Michela Ansuinelli, MD, PhD1,7*, Yohei Arihara, MD, PhD8*, Rao Prabhala, PhD1,2*, Carol G. Reynolds, PhD1*, Robert Gentleman, PhD3*, Serena De Vita, MD, PhD4*, Nikhil C. Munshi, MD2,9 and Jerome Ritz, MD1,2

1Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA
2Harvard Medical School, Boston, MA
3Center for Computational Biomedicine, Harvard Medical School, Boston
4Novartis Biomedical Research, Cambridge, MA
5Novartis Biomedical Research, Basel, Switzerland
6Novartis Pharmaceuticals Corporation, East Hanover, NJ
7Hematology, Department of Translational and Precision Medicine, Sapienza University of Rome, Rome, Italy
8The Cancer Institute Hospital of Japanese Foundation for Cancer Research, Tokyo, Japan
9Dana-Farber Cancer Institute, The Jerome Lipper Multiple Myeloma Center, Boston, MA

T-ChargeTM is a novel platform for manufacturing CAR-T cells that reduces ex-vivo processing time to <2 days. A phase 1 clinical trial (NCT04318327) of durcabtagene autoleucel (PHE885) manufactured using the T-ChargeTM platform in relapsed/refractory multiple myeloma (R/RMM) has shown promising clinical activity. Previous correlative studies showed that T-ChargeTM manufacturing preserved T cell stemness in the CAR-T product and PHE885 expanded robustly with high TCR clonal diversity (Ikegawa et al., ASH 2023). CAR-T cells persisted for prolonged periods in some patients, allowing the detailed analysis of long-term persisting CAR-T cells.

Among 32 patients with R/RMM treated at our center, we analyzed samples from 25 patients who had been followed for at least six months. At six months, CAR-T cells remained detectable in peripheral blood in 12 of 25 patients (median 38.3%; range, 2.1-70.8% of CD3+) by flow cytometry. CAR-T cells persisted for 12 months in 7 of 14 patients (median 15.6%; range, 1.04-75.9% of CD3+). At 24 months after CAR-T infusion, two of four patients continued to have detectable CAR-T by flow cytometry (61% and 0.8% of CD3+, respectively).

CAR-T cells were predominantly CD4+ with effector memory phenotype at peak expansion, whereas the persisting CAR-T cells at later time points contained more less-differentiated T cells, including naïve and stem-like memory T cells. Mass cytometry revealed that the expression of activation, proliferation, and exhaustion markers on CAR-T cells declined after peak expansion. CAR-T TCR repertoire continued to be highly diverse 6-18 months after infusion and CAR-T cells present at later time points shared TCR clonotypes with those at peak expansion.

Next, we evaluated the anti-BCMA function of post-infusion CAR-T cells in 12 patients who had persistent CAR-T cells in peripheral blood for >6 months. CAR-T cells were isolated by cell sorting from cryopreserved PBMC obtained during early expansion and at later time points (>3 months) and assayed their ability to directly kill reporter cells expressing BCMA. Isolated CAR-T cells continued to have anti-BMCA-specific cytotoxic activity that was not impaired at late time points. Notably, anti-BCMA-specific cytotoxicity in vitro was comparable in CAR T cells obtained from patients with ongoing clinical response and those with disease relapse. sBCMA levels at disease relapse were very low in 6 of 7 patients with CAR-T persistence. In contrast, sBCMA levels increased in 4 of 8 patients who relapsed after loss of CAR-T cells. These results indicate that T-chargeTM manufacturing produced CAR-T cells capable of long-term persistence with sustained anti-BCMA specific cytotoxic activity and BCMA loss likely contributes to relapse in these patients.

We also explored various factors that could affect CAR-T persistence. The number of CAR-T cells at peak expansion and the number of Tscm/central memory T cells in the CAR-T product/leukapheresis were not associated with long-term CAR-T persistence. CD127 (IL-7Ra) expression on CAR-T cells was significantly higher in patients with long-term CAR-T persistence in CD4 T cells (day14; p = 0.04, day21; p = 0.01, and day28; p = 0.02) and CD8 T cells (day14; p = 0.04 and day21; p = 0.005), and this difference was not observed for non-CAR-T cells. Mass cytometry data confirmed that the CAR-T subset with high expression of CD127 at peak expansion was significantly increased in patients with long-term CAR-T persistence (p = 0.03).

Because the T-chargeTM platform reduces ex-vivo CAR-T expansion, PHE885 expands predominantly in vivo. In conventional CAR-T manufacturing, common g-chain receptor cytokines, especially IL-7 and IL-15, promote ex-vivo expansion and long-term CAR-T persistence. Similarly, baseline cytokine levels may have an important role in the memory formation of post-infusion PHE885. Among common g-chain receptor cytokines, IL-7 levels at CAR-T infusion were significantly higher in patients with long-term CAR-T persistence (p = 0.03).

In summary, BCMA CAR-T cells manufactured using the T-Charge process exhibit robust expansion in vivo and long-term polyclonal persistence for >6 months in 48% of patients. Long-term persisting CAR-T cells remain functionally active against BCMA expressing target cells in vivo and in vitro, and the IL-7/IL-7R pathway appears to have an important role in promoting long-term persistence of memory CD4+ CAR-T cells.

Disclosures: Sperling: Novartis: Consultancy. Quinn: Novartis: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months. Bu: Novartis: Current Employment, Patents & Royalties: Novartis. Mataraza: Novartis: Current Employment. Pearson: Novartis: Current Employment, Current equity holder in publicly-traded company. Charles: Novartis: Current Employment. Credi: Novartis: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months. Orwitz: Novartis: Current Employment. Gentleman: 23AndMe: Current equity holder in publicly-traded company; Scorpion Tx: Current holder of stock options in a privately-held company, Membership on an entity's Board of Directors or advisory committees; Exai Tx: Current holder of stock options in a privately-held company; Boimap USA: Consultancy. Vita: Novartis: Current Employment. Munshi: AbbVie, Adaptive Bio, Amgen, Bristol Myers Squibb, Celgene, GlaxoSmithKline, Janssen, Karyopharm, Legend Bio, Novartis, Oncopep, Pfizer, Recordati, Sebia, Takeda: Consultancy; Oncopep: Current holder of stock options in a privately-held company. Ritz: Kite/Gilead: Research Funding; Novartis: Research Funding; Oncternal: Research Funding; Oncternal: Research Funding; Clade Therapeutics: Membership on an entity's Board of Directors or advisory committees; Garuda Therapeutics: Membership on an entity's Board of Directors or advisory committees; LifeVault Bio: Membership on an entity's Board of Directors or advisory committees; Smart Immune: Membership on an entity's Board of Directors or advisory committees; TriArm Bio: Membership on an entity's Board of Directors or advisory committees.

*signifies non-member of ASH