Single cell RNA sequencing to study the in vivo behaviour of allogeneic HSC-derived MCAR-engineered NKT cells

Ovarian cancer (OC) poses a significant challenge due to frequent recurrence linked to tumor heterogeneity and an immunosuppressive tumor microenvironment (TME). Here, we generated allogeneic MSLN-targeting CAR (MCAR)-engineered invariant natural killer T (MCAR-NKT) cells, demonstrating high-yield and high-purity production, potent antitumor efficacy, multiple tumor targeting mechanism, and significant in vivo expansion and persistence. Two therapeutic cell products (i.e., CAR-T and CAR-NKT cells) were analyzed at five time points: day 0 (pre-infusion), and days 14, 28, 42, and 56 post-infusion. Day 0 samples were collected prior to cell administration, whereas post-infusion samples were isolated from tumor sites, peripheral blood, spleen, and liver. These findings underscore the feasibility and therapeutic potential of allogeneic MCAR-NKT cell products for OC, laying a foundation for further translation and clinical development.