Translating a Stem Cell-Based Therapy for Epidermolysis Bullosa into the Clinic
Abstract
The proposal develops a stem-cell based therapy for recessive dystrophic epidermolysis bullosa (RDEB), which is one of the most severe forms of epidermolysis bullosa (EB), a group of rare inherited skin blistering diseases. To accomplish this goal, we have utilized genetic correction of patient-specific induced pluripotent stem cells (iPSCs) followed by the differentiation of these corrected iPSCs into epidermal cells and fibroblasts for the generation of composite full thickness skin grafts for transplantation. During the current award, we validated the reproducibility of our combined gene editing and reprogramming approach. We also produced and characterized genetically corrected RDEB iPSCs that we will use to continue the generation of IND-enabling data. We developed a cGMP-compliant procedure for the generation of these iPSCs in the Gates Biomanufacturing Facility (GBF). In addition, we developed a cGMP-compatible procedure for enriching keratinocytes derived from iPSCs (iKCs) using a cGMP-compliant CliniMACS Prodigy enrichment approach. We did face a problem of inconsistent engraftment of iKCs and iPSC-derived fibroblasts (iFBs). For this reason, we modified our protocols for the derivation of iKCs and iFBs. We now derive these cells from hair-bearing human skin organoids. We can now consistently generate hair bearing skin organoids from all our genetically corrected RDEB iPSCs. We also found that iKCs and iFBs isolated from these organoids show more consistent functionality as compared to iKCs and iFBs generated using our conventional monolayer protocol. The organoid-based differentiation protocol will now allow us to generate both genetically corrected RDEB iKCs and iFBs in one differentiation procedure followed by CliniMACS Prodigy enrichment using cell type specific surface markers. This will simplify and reduce the cost of manufacturing our stem cell therapy for EB.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 2024
- Accession Number
- AD1230037
Entities
People
- Dennis R. Roop
Organizations
- University of Colorado Boulder