Genome engineering therapeutics for cystinuria and its metabolic consequences.

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ABSTRACT Cystinuria is an inherited human kidney disease with significant morbidity affecting 1 in 7000, including veterans. The disease is caused by mutation of genes involved in renal cystine transport resulting in elevated urinary cystine with kidney stone formation. With the genetic basis of the disorder defined (mutation in SLC3A1, cystinuria type A), opportunities for targeted molecular therapies exist. Building upon our previous grant, we propose an innovative experimental design to demonstrate long-term phenotypic correction of cystinuria in an intact animal using a combination of transposon and adeno-associated virus (AAV) technologies. In specific aim 1, we will test the ability of hybrid AAV-piggyBac transposon technology to mediate long-term transgene expression in mouse kidney and correct cystinuria in a Slc3a1-/- mouse model. In specific aim 2, we will test the ability of novel proximal tubule targeted AAV to deliver transgenes to proximal tubule cells in mice in vivo. In both aims, we will test delivery to and evaluate for long-term transgene expression in both neonatal and adult mice. In specific aim 3, we propose to bridge our foundational pre-clinical studies to human application by testing AAV delivery to human kidney organoids including cystinuria organoids. Our innovative but feasible studies will expand use of AAV and transposon technology for kidney gene delivery and lay a pre-clinical foundation for ultimate human therapeutic application for kidney disease in veterans and others.