Renee Cottle, PhD, Clemson University
PROJECT TITLE: Ex vivo CRISPR-Cas9 induced gene correction of inborn errors of liver metabolism
Inherited metabolic liver disease is characterized by the aberrant synthesis of enzymes or other proteins essential for proper metabolism in the liver, which occurs in one in one thousand births. Liver transplantation is the only definitive therapy available for treatment of metabolic liver disorders, but it has severe drawbacks. We aim to develop new cell-based therapies for inherited metabolic liver disorders using gene-editing tools to correct the disease at the genomic level in the patient’s hepatocytes.
Homozygous familial hypercholesterolemia (HoFH) is characterized by impaired LDL metabolism in the liver caused by mutations in the LDL receptor (LDLR) gene, the loss of which results in the rapid development of severe cardiovascular disease as early as childhood. Although the only curative therapy for severe HoFH, allogeneic liver transplantation is not a standard therapy for HoFH because of related complications. Clustered regularly interspaced short palindromic repeats (CRISPR) and associated Cas nucleases has unleashed new possibilities for HoFH and other inherited metabolic liver disorders. A novel curative approach involves CRISPR-Cas induced homologous directed repair (HDR) for precise insertion of a therapeutic transgene in the patient’s hepatocytes. One challenge of this approach is that corrected hepatocytes lack a natural selective advantage over mutant cells for repopulation of the liver. Therefore, fixing the LDLR deficiency is not enough. We must also provide a competitive advantage in gene corrected hepatocytes over native hepatocytes for a repopulation of the liver. The overall objective of this proposal is to develop a novel strategy for gene modification in primary hepatocytes.
Developmental Research Project Program (DRP) funding through SC INBRE has enabled our laboratory to purchase the materials and supplies as well as equipment for this work. Dr. Cottle, the project PI, has had access to career and research mentoring from established PIs in her field as a result of her participation in the DRP program.
During the funding period, we established protocols for culturing and transfection into primary hepatocytes ex vivo. We designed gene editing tools for targeting a therapeutic gene that will providing hepatocytes a competitive advantage for the repopulation of the liver. These advances have placed us closer to publishing high impact articles on gene editing in primary hepatocytes as a novel therapeutic strategy for metabolic liver disease.
Publications as a result of SC INBRE funding for this project include:
- Bryson TE, Anglin CM, Bridges PH, Cottle RN. Nuclease-mediated gene therapies for inherited metabolic diseases of the liver. YJBM. Submitted for publication (December 2017).
January 12, 2018