Unlocking the Potential of Allogeneic Vd2 T Cells for Ovarian Cancer Therapy through CD16 Biomarker Selection and CAR/IL-15 Engineering

Allogeneic V?9Vd2 (Vd2) T cells are attractive candidates in the development of cancer therapies due to their demonstrated safety in allogeneic settings and innate ability to fight tumors. However, the limited clinical success of Vd2 T cell-based treatments may be attributed to donor variability, short-lived persistence, and tumor evasion. To address these limitations, we have generated Vd2 T cells with improved properties. By utilizing CD16 as a donor selection biomarker, we have generated Vd2 T cells with high levels of cytotoxicity and potent antibody-dependent cell-mediated cytotoxicity (ADCC) function, and RNA sequencing characterization supports the increased effector function of Vd2 T cells obtained from CD16 high (CD16Hi) donors. Further improvement was achieved through CAR and IL-15 engineering techniques. Preclinical studies in two ovarian cancer models showed that engineered CD16Hi Vd2 T cells are both effective and safe, targeting tumors through multiple mechanisms, exhibiting long-term persistence in vivo, and not causing graft-versus-host disease. These findings support the potential of engineered CD16Hi Vd2 T cells as a viable cancer therapy option. Overall design: To assess the differences between Vd2 T cells that carry different CD16 expression levels, we performed bulk RNA-Seq on Vd2 T cells expanded from 13 PBMC donors (all Vd2 T cells were cultured and expanded in the same way). In this study, 7 new Vd2 T cell samples were sequenced, and 6 Vd2 T cell samples were sequenced in our previous study (GEO Accession: GSE164425), thus we downloaded the raw sequencing data and reanalyzed them. Besides the Vd2 T cell samples, we also sequenced 4 new NK cell samples and retrieved 3 abT cell samples from our previous study (GEO Accession: GSE164425) as a reference. All the samples were subject to the same processing and analysis procedure as denoted in the PROTOCOLS section. Re-analysed Samples PRJNA690603/GSE164425: GSM5009488 SRR13381427/SRX9804343 SAMN17255257 GSM5009489 SRR13381428/SRX9804344 SAMN17255248 GSM5009490 SRR13381429/SRX9804345 SAMN17255241 GSM5009491 SRR13381430/SRX9804346 SAMN17255231 GSM5009492 SRR13381431/SRX9804347 SAMN17255222 GSM5009493 SRR13381432/SRX9804348 SAMN17255214 GSM5009496 SRR13381435/SRX9804351 SAMN17255123 GSM5009497 SRR13381436/SRX9804352 SAMN17255122 GSM5009499 SRR13381438/SRX9804354 SAMN17255120