Natural killer (NK) cells are innate immune cells comprising 5–10% of peripheral blood (PB) lymphocytes. As key players in innate immunity, these cytotoxic cells hold significant promise for cancer treatment. However, current NK cell therapies based on primary NK cells face considerable challenges, including scalability, consistency, donor variability, and safety concerns associated with gene-modified NK cells aimed at enhancing anti-tumour activity. The yield and quality of primary NK cells also vary widely among donors.

In contrast, NK cells derived from induced pluripotent stem cells (iNK cells) present a scalable, off-the-shelf alternative to overcome these limitations. Induced pluripotent stem cells (iPSCs), generated by reprogramming somatic cells into a pluripotent state, can differentiate into any cell type, including NK cells. iPSC-derived NK cells provide a homogeneous, reproducible, and unlimited source of NK cells, eliminating donor variability, ensuring consistent quality, and enabling multi-dose treatment regimens. Clinical trials using iPSC-derived NK cells have received FDA approval and are ongoing, highlighting their potential as uniform, highly cytotoxic NK cell products for cancer immunotherapy. Despite their promise, challenges persist. There is no universal protocol effective across iPSCs from various somatic sources, the role of transcription factors in iNK development remains unclear, and while genetic modifications enhance anti-tumour activity, they raise safety concerns.

My research addresses these challenges by establishing a universal iNK differentiation protocol with rapid ex vivo expansion. iNK cells are collected at different developmental stages, and single-cell RNA sequencing is performed to evaluate the role of transcription factors. Additionally, the project aims to develop a robust and safe CRISPR/Cas9-AAVS1 genome knock-in platform to enhance the anti-tumour capabilities of iNK cells. These outcomes promise to advance iNK cell therapy by overcoming current obstacles, facilitating clinical trials, and accelerating the development and approval of iNK-based cancer immunotherapies.