AKT inhibition generates potent polyfunctional clinical grade AUTO1 CAR T-cells, enhancing function and survival

Area of Science:

• Immunotherapy
• Cellular Therapy
• Oncology

Background:

• AUTO1 is a CD19-targeting chimeric antigen receptor (CAR) T-cell therapy effective in adult lymphoblastic leukemia.
• Optimal CAR-T cell efficacy relies on Tscm/Tcm cell populations, which are scarce in heavily pretreated adult patients.
• Strategies are needed to enhance Tscm/Tcm cell numbers and function in CAR-T cell manufacturing.

Purpose of the Study:

• To evaluate the use of an AKT inhibitor (VIII) to uncouple T-cell expansion from differentiation.
• To enrich Tscm/Tcm subsets within AUTO1 CAR-T cell products.
• To assess the impact of VIII on AUTO1 cell function and manufacturing scalability.

Main Methods:

• Incorporation of AKT inhibitor (VIII) into the AUTO1 manufacturing process using the CliniMACS Prodigy platform.
• Evaluation of Tscm/Tcm enrichment, cell expansion, cytotoxicity, and polyfunctionality in vitro.
• Assessment of in vivo anti-tumor activity and metabolic profiles.
• cGMP scale manufacturing validation.

Main Results:

• AUTO1 manufactured with VIII demonstrated significant Tscm/Tcm enrichment, improved expansion, and enhanced cytotoxicity in vitro.
• VIII-treated AUTO1 cells exhibited superior anti-tumor activity in vivo, Th1/Th17 skewing, and increased polyfunctionality.
• A unique metabolic profile and autophagy signature were observed, supporting enhanced cell function.
• VIII-based manufacturing is scalable to cGMP levels and yields AUTO1 products with superior phenotype and function.

Conclusions:

• The use of AKT inhibitor (VIII) during manufacturing enhances AUTO1 CAR-T cell product quality and anti-tumor efficacy.
• VIII promotes Tscm/Tcm enrichment and improves key functional parameters, addressing limitations in heavily pretreated patients.
• This approach may help overcome product-related factors contributing to CD19+ relapse in lymphoblastic leukemia.