Development of a targeted All-in-One Delivery Vehicle (AiDV) for delivery of gene editing Homology-Directed Repair (HDR) components to specific cells in vivo to treat inborn error of immunity

The end goal of the project is to efficiently deliver Cas9 and a cDNA of a gene to integrate the cDNA site specifically by homology-directed repair (HDR) to treat inborn errors of immunity.

I intend (aim 1) to bring state-of-the-art Cas9 HDR technology to Sweden to treat X-linked agammaglobulinemia (XLA), based on ex vivo electroporation of Cas9 and subsequent delivery of the cDNA by adeno-associated virus 6 (AAV6) developed by my collaborator at UCLA.

In Aims 2&3, the HDR delivery will be further improved by developing a novel targeted nanoparticle delivering Cas9 and the cDNA template to specific cells in vivo.

Thus, delivering the entire HDR machinery by a single nanoparticle to only hematopoietic stem cells (HSCs) or T-cells in vivo. I will leverage my expertise in extracellular vesicles (EVs) to create an All-in-one Delivery Vehicle (AiDV).

The AiDV will be more efficient and less toxic than the electroporation/AAV6 technique and allow for the development of in vivo delivery of the HDR machinery.

The project will benefit patients in a short perspective (Aim 1), by bringing HDR gene editing closer to a clinical trial, and in the long perspective by improving the treatment with the final goal of developing an in vivo treatment, which would revolutionize the treatment of primary immunodeficiencies.

Hence, the project is translational and will further our arsenal to treat HSC genetic diseases and advance the ATMP and precision medicine fields at Karolinska, and Sweden.