Cell-based analysis of CAD variants identifies individuals likely to benefit from uridine therapy.

PURPOSE: Pathogenic autosomal recessive variants in CAD, encoding the multienzymatic protein initiating pyrimidine de novo biosynthesis, cause a severe inborn metabolic disorder treatable with a dietary supplement of uridine. This condition is difficult to diagnose given the large size of CAD with over 1000 missense variants and the nonspecific clinical presentation. We aimed to develop a reliable and discerning assay to assess the pathogenicity of CAD variants and to select affected individuals that might benefit from uridine therapy. METHODS: Using CRISPR/Cas9, we generated a human CAD-knockout cell line that requires uridine supplements for survival. Transient transfection of the knockout cells with recombinant CAD restores growth in absence of uridine. This system determines missense variants that inactivate CAD and do not rescue the growth phenotype. RESULTS: We identified 25 individuals with biallelic variants in CAD and a phenotype consistent with a CAD deficit. We used the CAD-knockout complementation assay to test a total of 34 variants, identifying 16 as deleterious for CAD activity. Combination of these pathogenic variants confirmed 11 subjects with a CAD deficit, for whom we describe the clinical phenotype. CONCLUSIONS: We designed a cell-based assay to test the pathogenicity of CAD variants, identifying 11 CAD-deficient individuals who could benefit from uridine therapy

Degradation of subcells and tunnel junctions during growth of GaInP/Ga(In)As/GaNAsSb/Ge 4-junction solar cells

A GaInP/Ga(In)As/GaNAsSb/Ge 4J solar cell grown using the combined MOVPE + MBE method is presented. This structure is used as a test bench to assess the effects caused by the integration of subcells and tunnel junctions into the full 4J structure. A significant degradation of the Ge bottom subcell emitter is observed during the growth of the GaNAsSb subcell, with a drop in the carrier collection efficiency at the high energy photon range that causes a ~15% lower Jsc and a Voc drop of ~50 mV at 1-sun. The Voc of the GaNAsSb subcell is shown to drop by as much as ~140 mV at 1-sun. No degradation in performance is observed in the tunnel junctions, and no further degradation is neither observed for the Ge subcell during the growth of the GaInP/Ga(In)As subcells. The hindered efficiency potential in this lattice-matched 4J architecture due to the degradation of the Ge and GaNAsSb subcells is discussed. (C) John Wiley & Sons Lt