Correcting APDS2 syndrome-caused mutation for future gene and cell therapies

I Romayor(1,2) M Inglés-Ferrándiz(1,2) M Martin-Inaraja(1,2) L Herrera(1,2) S Santos(1,2) M A Vesga(1,2) J Anguita(3) L M Allende(4,5) L I Gonzalez-Granado(4,5) C Eguizabal(1,2)

1:BioCruces/Hospital Universitario Cruces; 2:Centro Vasco de Transfusión y Tejidos Humanos (CVTTH); 3:CIC bioGUNE; 4:Hospital Universitario 12 de Octubre; 5:Universidad Complutense de Madrid

APDS2 syndrome is a genetic disorder classified as a primary immunodeficiency and caused by mutations in PIK3R1 gene. Such alteration results in a dysfunctional immune system, affecting NK cells among others. Giving the lack of specialized treatments against APDS2 syndrome, we aim to develop novel strategies to uncover effective specific-therapies directed to APDS2 patients by correcting the mutation in their own cells. For this purpose, we will use the CRISPR/Cas9 molecular tool to edit the genome of an APDS2-derived iPS cell line recently generated in our laboratory. According to the characteristics of the iPS cells, we will utilize nucleofection as the transfection method for CRISPR/Cas9 complex internalization, which supplies a less aggressive and more reproducible gene editing technology. Finally, we will employ corrected APDS2-derived iPS to differentiate them into hematopoietic stem cells expressing the CD34+ marker. These hematopoietic progenitors could be also differentiated into the different types of blood cells, including NK cells. To this extent, we will be able to obtain not only normal NK cells, but also a personal source of healthy transfusable components from the APDS2 patient. As a whole, our work represents an innovative therapy that will improve current treatments, providing an exceptional and powerful tool to study APDS2 syndrome as well as personalized gene and cell therapy. Moreover, our findings could be extrapolated to the research of similar pathologies, facilitating the development of advanced targeted gene and cell therapies.