Genome engineering therapeutics for cystinuria and its metabolic consequences.

胱氨酸尿症的基因组工程疗法及其代谢后果。

基本信息

批准号：
10588590
负责人：
MATTHEW H WILSON
金额：
--
依托单位：
VETERANS HEALTH ADMINISTRATION
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-04-01 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10588590
关键词：
Acute Adult Affect Amino Acids Animal Model Animals Blood Cells Chronic Chronic Kidney Failure Clinical Cystine Cystinuria Data Dependovirus Development Disease Disease model Experimental Designs Feasibility Studies Foundations Gene Delivery Gene Mutation Gene Transfer Genetic Genetic Recombination Genome engineering Glutathione Grant Health Human Hybrids Image Inherited Injury Injury to Kidney Kidney Kidney Calculi Kidney Diseases Knowledge Mediating Medical Metabolic Metabolic Pathway Mission Modeling Modification Morbidity - disease rate Mus Mutation Neonatal Organ Organoids Phenotype Plague Plasma Proximal Kidney Tubules Recombinants Research Site Sodium Chloride Technology Technology Transfer Testing Therapeutic Tissues Trans-Splicing Transgenes Urine Validation Veterans Water absorption adeno-associated viral vector cellular transduction in vivo innovation mass spectrometric imaging military veteran molecular targeted therapies mortality mouse model neutralizing antibody novel novel therapeutics pre-clinical preclinical study prevent reconstitution side effect transgene delivery transgene expression translational therapeutics urinary vector

项目摘要

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.

抽象的胱氨酸尿症是一种遗传性人类肾脏疾病，发病率高达七千分之一，其中包括退伍军人。该疾病是由参与肾脏胱氨酸转运的基因突变引起的，导致尿蛋白升高胱氨酸与肾结石形成有关。确定了该疾病的遗传基础（SLC3A1 突变， A 型胱氨酸尿症），存在靶向分子治疗的机会。在我们之前的资助的基础上，我们提出了一种创新的实验设计，以证明胱氨酸尿症的长期表型校正使用转座子和腺相关病毒（AAV）技术相结合的完整动物。在具体目标1中，我们将测试混合AAV-piggyBac转座子技术介导长期的能力小鼠肾脏中的转基因表达并纠正 Slc3a1-/- 小鼠模型中的胱氨酸尿症。在具体目标 2 中，我们将测试新型近端小管靶向 AAV 将转基因传递至小鼠近端小管细胞的能力体内。在这两个目标中，我们将测试新生儿和新生儿的长期转基因表达并评估其长期转基因表达。成年小鼠。在具体目标 3 中，我们建议通过以下方式将我们的基础临床前研究与人类应用联系起来：测试 AAV 向人类肾脏类器官（包括胱氨酸尿类器官）的传递。我们的创新但可行研究将扩大 AAV 和转座子技术在肾脏基因传递中的应用，并奠定临床前基础为退伍军人和其他人的肾脏疾病的最终人类治疗应用奠定了基础。