Point-of care process transfer for GMP-grade manufacturing of ex-vivo expanded and activated γδ T Cells (EAGD) following haploidentical bone marrow transplantation and post-BMT cyclophosphamide

Abstract

Background & Aim Background Previous studies show that tumor infiltrating gamma delta (γδ) T cells are linked to improved survival in solid tumors and that high numbers of circulating γδ T cells are linked to improved overall survival in hematopoietic malignancies, both suggesting that γδ T cell-based therapies could improve patient outcomes. This report describes the manufacturing optimization for our first-in-human Phase I trial of haploidentical allogeneic hematopoietic cell transplantation (HCT), post-HCT Cy, and subsequent infusion of expanded/activated γδ T cells (EAGD) (NCT03533816). The aim of this study was to determine whether an industry-academic process transfer could be successfully executed and carried out in a point-of-care (POC) setting. Methods, Results & Conclusion Methods The EAGD process was co-developed by the University of Alabama at Birmingham and Incysus Therapeutics, Inc. for the Miltenyi CliniMACS Prodigy®, a commercially available, automated closed-system bioreactor readily adaptable to POC cell manufacturing. Process transfer and GMP-grade manufacturing optimization was conducted at the Midwest Stem Cell Therapy Center (MSTCT) at the University of Kansas, the initial clinical trial site. Four manufacturing qualification procedures were conducted. Mononuclear cell preparation was followed by γδ T cell culture in OpTmizer® media + Zoledronate and Interleukin-2 (ZOL/IL-2). Cell number, viability, and lymphocyte phenotype were monitored until an infusible γδ T cell dose was ascertained at which time immunomagnetic αβ T cell depletion was performed and final product harvested and cryopreserved. Results ZOL/IL-2 EAGD manufacturing at MSCTC resulted in γδ T cell enrichment from 3.0% to 91.0% ±12.0% (66.5% to 95.0%) with a final viability of >90%. All products met criteria for a minimal infusible dose of 1.0 × 106 γδ T cells/kg and a maximum final αβ T cell concentration of Conclusions The EAGD process was successfully transferred from Incysus Therapeutics to the MSCTC thereby enabling POC manufacturing for the clinical protocol at the treating institution. Although the challenges of donor-to-donor γδ T cell expansion variability and the critical timing for this protocol remain, our experience represents the integrated effort of industry and academic settings to facilitate the potential pharmacoeconomic impact for bedside delivery of cell therapy products.