![]() ![]() The blood sample of a Bckdha −/− mice was insufficient for analysis. Source data are provided as a Source data file.Ī Weight curves and b plasma leucine concentrations at sacrifice ( Bckdha −/− injected with 10 13 vg/kg and sacrificed <4 weeks n = 2 (2 females), Bckdha −/− injected with 10 13 vg/kg and sacrificed at 4 weeks n = 5 (1 male/4 females), Bckdha −/− injected with 10 14 vg/kg and sacrificed at 4 weeks n = 2 (1 male/1 female), Bckdha +/− n = 18 (9 males/9 females), Bckdha +/+ n = 9 (2 males/7 females). d Western blot analyses of human BCKDHA in Huh7 cells transfected with hAAT GFP, hAAT iSV40 h BCKDHA WT, hAAT iSV40 h BCKDHA co1 (codon-optimized sequence 1), and hAAT iSV40 h BCKDHA co2 (codon-optimized sequence 2), the three of them carrying the SV40 intron ( n = 3 biological replicates for each condition). c AAV8-hAAT-h BCKDHA vector genome consisting of an expression cassette including hepatocyte-specific hAAT promoter composed of the ApoE enhancer and the human alpha 1 antitrypsin promoter (hAAT), a SV40 intronic sequence, the h BCKDHA coding sequence, a WPRE sequence and a polyadenylation signal (bGH poly(A)), flanked by AAV2 ITRs. b Western blot analyses of human BCKDHA in Huh7 cells transfected with EF1α GFP, EF1α h BCKDHA WT, EF1α h BCKDHA co1 (codon-optimized sequence 1) and EF1α h BCKDHA co2 (codon-optimized sequence 2) ( n = 3 biological replicates for each condition). ![]() Here we show efficacy of gene therapy for MSUD demonstrating its potential for clinical translation.Ī AAV8-EF1α-h BCKDHA vector genome consisting of an expression cassette including a ubiquitous human elongation factor 1-alpha promoter (hEF1a) with an extra intronic sequence, the h BCKDHA coding sequence, a WPRE sequence, and a polyadenylation signal (bGH poly(A)), flanked by AAV2 ITRs. While the use of a ubiquitous promoter fully and sustainably rescued the disease (long-term survival, normal phenotype and correction of biochemical abnormalities), liver-specific expression of BCKDHA led to partial, though sustained rescue. BCKDHA gene transfer rescued the lethal phenotype. Animals were treated at P0 with intravenous human BCKDHA AAV8 vectors under the control of either a ubiquitous or a liver-specific promoter. We establish and characterize the Bckdha (branched chain keto acid dehydrogenase a) -/- mouse that exhibits a lethal neonatal phenotype mimicking human MSUD. Here, based on significant liver contribution to the catabolism of the branched chain amino acid leucine, we develop a liver-directed adeno-associated virus (AAV8) gene therapy for MSUD. Treatment for MSUD represents an unmet need because the current treatment with life-long low-protein diet is challenging to maintain, and despite treatment the risk of acute decompensations and neuropsychiatric symptoms remains. syrup urine disease (MSUD) is a rare recessively inherited metabolic disorder causing accumulation of branched chain amino acids leading to neonatal death, if untreated. 13 Inserm UMR_S1163, Institut Imagine, Paris, France.
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