APDS2 syndrome is a primary immunodeficiency caused by mutations in PIK3R1 gene. Such genetic alterations affect to immune cells, resulting in an immunologic dysfunction. To date, the only available treatment against APDS2 disease consists in hematopoietic stem cells transplantation, bringing poor efficacy and many adverse effects. For this reason, our aim is to develop novel gene and cell strategies to uncover effective specific-therapies directed to APDS2 patients. For this purpose, we use CRISPR/Cas9 molecular tool for genetic edition to correct the PIK3R1 mutation in the genome of an APDS2-derived iPSC line generated in our laboratory.

Results show that our nucleofected CRISPR/Cas9 complex present an efficiency of a 60% without off-target effects or chromosome aberrations. The corrected clones maintain their identity as pluripotent stem cells and are able to differentiate towards the three germ layers more efficiently compared to the APDS2-mutated cells. Furthermore, our clones are able to be driven towards CD34+ hematopoietic progenitors, obtaining functional immune cell precursors from the APDS2 patient. Finally, our corrected clones show a better differentiation potential towards the hematopoietic lineage respect to the mutated cells.

Taking into account such findings, our work represents an innovative therapy that could improve current treatments, providing an exceptional and powerful tool to study APDS2 syndrome as well as personalized gene and cell therapy. Moreover, our results could be extrapolated to the research of similar pathologies, facilitating the development of advanced targeted gene and cell therapies.