Demonstrating the Manufacture of Human CAR-T Cells in an Automated Stirred-Tank Bioreactor.

Elena Costariol,Arman Amini,Thomas R J Heathman,Christopher J Hewitt,Alvin W Nienow,Qasim A Rafiq,Marco C Rotondi

doi:10.1002/biot.202000177

Abstract

Chimeric antigen receptor T-cell (CAR-T) therapies have proven clinical efficacy for the treatment of hematological malignancies. However, CAR-T cell therapies are prohibitively expensive to manufacture. The authors demonstrate the manufacture of human CAR-T cells from multiple donors in an automated stirred-tank bioreactor. The authors successfully produced functional human CAR-T cells from multiple donors under dynamic conditions in a stirred-tank bioreactor, resulting in overall cell yields which were significantly better than in static T-flask culture. At agitation speeds of 200 rpm and greater (up to 500 rpm), the CAR-T cells are able to proliferate effectively, reaching viable cell densities of >5 × 106 cells ml-1 over 7 days. This is comparable with current expansion systems and significantly better than static expansion platforms (T-flasks and gas-permeable culture bags). Importantly, engineered T-cells post-expansion retained expression of the CAR gene and retained their cytolytic function even when grown at the highest agitation intensity. This proves that power inputs used in this study do not affect cell efficacy to target and kill the leukemia cells. This is the first demonstration of human CAR-T cell manufacture in stirred-tank bioreactors and the findings present significant implications and opportunities for larger-scale allogeneic CAR-T production.

Highlights

Chimeric antigen receptor T-cell (CAR-T) therapies have proven clinical efficacy for the treatment of hematological malignancies
Do not affect cell efficacy to target and kill the leukemia cells. This is the first demonstration of human CAR-T cell manufacture in stirred-tank bioreactors and the findings present significant implications and opportunities for larger-scale allogeneic CAR-T production
This study demonstrated, for the first time, the expansion of CAR-T cells in a STR

Summary

Introduction

Chimeric antigen receptor T-cell (CAR-T) therapies have proven clinical efficacy for the treatment of hematological malignancies. At agitation speeds of 200 rpm and greater (up to 500 rpm), the CAR-T cells are able to proliferate cell therapies in particular have demonstrated significant clinical efficacy for conditions such as acute lymphoblastic leukemia and non-Hodgkin’s lymphoma and have received US Food and Drug Administration (FDA) and European Medicines Agency effectively, reaching viable cell densities of >5 × 106 cells ml-1 over 7 days This is comparable with current expansion systems and significantly better than static expansion platforms (T-flasks and gas-permeable culture bags). Engineered T-cells post-expansion retained expression of the CAR gene and retained their cytolytic function even when grown at the highest agitation intensity This proves that power inputs used in this study (EMA) regulatory approval (e.g., Novartis’ Kymriah and Gilead’s Yescarta).[1] Despite the positive clinical outcomes, these patient-specific therapies are expensive to manufacture, cost in excess of $250 000 and their current manufacture involves open, manual processes.[2] To reduce the cost, do not affect cell efficacy to target and kill the leukemia cells. This is the first demonstration of human CAR-T cell manufacture in stirred-tank bioreactors and the findings present significant implications and opportunities for larger-scale allogeneic CAR-T production

Results

Discussion

Conclusion