Research Projects

Beta-thalassemia and HBE are genetic disorders that hinder the production of functional beta hemoglobin, causing moderate to severe anemia, hepatosplenomegaly, and other secondary complications. These conditions reduce quality of life and shorten life expectancy. Allogenic stem cell transplant is the only curative therapy, but it is often impeded by the lack of HLA-matched donors. Advances in genome engineering have opened up new therapeutic opportunities for beta-thalassemia and HbE. Direct correction of HBE mutations can reduce disease severity and improve transfusion dependency. Current gene editing approaches rely on programmable nuclease-mediated homology-directed repair (HDR), which is inefficient and generates unwanted insertions or deletions. Base editing allows for the creation of mutations without causing double-strand breaks (DSB). This proposal aims to correct the highly prevalent HbE and CD 15 (G-A) hemoglobin variant point mutations in the beta-globin gene using a base editor. Guide RNAs will be designed to correct these mutations using a base editor with minimal indels and without large deletions. The validated gRNAs and base editor mRNA will be used in CD34+ HSPCs derived from HBE/beta-thalassemia patients for in-vitro correction. After confirming editing at target sites, edited CD34+ HSPCs will be differentiated into erythroid cells and functional gene reconstitution will be validated at RNA and protein levels using RT-PCR and HPLC. The long-term multilineage engraftment potential of the base edited CD34+ HSPCs will be determined in immunocompromised NBSGW mice.