Dissecting A-to-I RNA Editome of Liver Macrophages in Non-Alcoholic Fatty Liver Disease

pup growth, serum BCAA levels through mass spectrometry, DBT protein expression via immunoblotting and vector genome copy via digital droplet PCR.Immunohistochemistry was used to assess presence of DBT protein in liver sections.As neonatal vector administration improved survival but did not rescue all pups, the timing of vector delivery was suspected to be critical, and the same vector was administered in utero to pups on E15.Equivalent studies are being initiated to compare outcomes following in utero treatment.Results: Gene addition with a codon optimised DBT human transgene significantly improved survival, growth and BCAA levels ( p < 0.05) compared to untreated knockout pups when administered on D0 of life.DBT protein in vector treated homozygous knock out pups was expressed at 2.5 times higher than untreated wild type pups.Immunohistochemistry demonstrated presence of DBT protein in vector treated pups (Figure 1).We will also present the results from the in utero vector administration.Figure 1.Presence of DBT protein ( purple) in murine liver cells in A) untreated DBT -/-and B) D0 vector treated DBT -/-mouse pups.Hepatocyte nuclei were co-stained (blue, DAPI).Conclusion: Neonatal murine liver transduction with a human DBT transgene significantly improves the disease phenotype in a murine neonatal lethal model of MSUD.Liver directed AAV mediated gene therapy represents a potential novel treatment for MSUD.