Develop AAV9 gene replacement therapy for treating MPS I

项目摘要 NeuroGT，Inc.是由基因疗法副教授傅海燕博士创立的初创公司 位于教堂山的北卡罗来纳大学中心，其使命是有效地开发和商业化 治疗人类罕见神经遗传性疾病的基因治疗产品。这个项目的目标是开发一种 针对治本的有效基因治疗产品 粘多糖症(MPS)I是一种由常染色体隐性遗传引起的致死性溶酶体病 α-L-艾杜糖苷酶缺陷。严重形式的MPS I(MPS IH，Hurler综合征)占大多数 已知病例，通常在10岁之前过早死亡，主要是由于神经系统 病情恶化和心肺衰竭。目前还没有有效的治疗方法来治疗神经系统疾病的适应症 下院议员IH。由于全局性弥漫性神经病变和血脑屏障(BBB)，MPS IH并不顺从 重组酶替代疗法或骨髓移植，这是治疗的标准 治疗MPS躯体症状的护理I.针对根本原因的基因替代疗法 已被证明是治疗单基因疾病的理想策略。大量研究表明， 成功地静脉或鞘内(IT)注射反式BBB-嗜神经AAV9用于治疗神经遗传性疾病。 重要的是，静脉注射和IT rAAV9的有效性和安全性已经被证明是非常高的 可在包括LSD在内的不同神经遗传性疾病中重现。 为了优化治疗潜力，我们开发了一种新型的自互补(Sc)AAV载体，scAAV- MCMV-∆hIDUAop，构建了密码子优化的小型化人IDUA基因。当在体外测试时 在人MPS IH细胞中，该ScAAV-∆hIDUAop载体构建的表达载体在 Level 8-比单链(Ss)rAAV-hIDUAop载体高8倍。重要的是，转导与 ScAAV-∆hIDUAop载体还可导致rIDUA的分泌增加11倍，从而可进入非 转换MPS I细胞并清除那里的GAG存储。我们的初步数据有力地支持了 人类临床应用的进一步发展。在这个拟议的项目中，出于临床意义，我们 我将使用AAV9、Via IV、鞘内(IT)和IV+IT在MPS I小鼠模型中测试新的ScAAV-∆hIDUAop载体 以评估治疗潜力并确定治疗MPS的最佳方案。 拟议的项目将使我们能够生成严格的临床前疗效和安全性数据，以支持 随后的临床开发和商业化。

Project Summary NeuroGT, Inc is a start-up company founded by Dr. Haiyan Fu, an associate professor in the Gene Therapy Center at University of North Carolina at Chapel Hill, with the mission of develop and commercialize effective gene therapy products to treat rare neurogenetic diseases in humans. The goal of this project is to develop an effective gene therapy product targeting the root cause for treating Mucopolysaccharidosis (MPS) I is a fatal lysosomal storage disease (LSD) caused by autosomal recessive defects in α-L-iduronidase (IDUA). Severe form of MPS I (MPS IH, Hurler syndrome) represents the majority of known cases, with premature deaths usually before age 10 years, predominantly due to neurological deterioration and cardiorespiratory failure. No effective treatment is available for neurological indications of MPS IH. Because of the global diffuse neuropathy and the blood brain barrier (BBB), MPS IH is not amenable to either recombinant enzyme replacement therapy or bone marrow transplantation, which are the standard of care for treating somatic symptoms of MPS I. Gene replacement therapy targeting the root cause has been demonstrated to be an ideal strategy for treating monogenic diseases. Numerous studies have demonstrated successful in IV or intrathecal (IT) delivery of trans-BBB-neurotropic AAV9 for treating neurogenetic diseases. Importantly, the efficacy and safety profiles of IV and IT rAAV9 delivery have been demonstrated to be highly reproducible across different neurogenetic diseases, including LSDs. For optimal therapeutic potential, we have developed a novel self-complementary (sc) AAV vector, scAAV- mCMV-∆hIDUAop, to deliver a miniaturized human IDUA cDNA with codon-optimization. When tested in vitro in human MPS IH cells, this scAAV-∆hIDUAop vector construct was shown to express functional IDUA protein at a level 8-fold higher, than the single-stranded (ss) rAAV-hIDUAop vector. Importantly, the transduction with scAAV--∆hIDUAop vector also lead to 11-fold increase in the secretion of rIDUA, which can enter non- transduce MPS I cells and clear GAG storage there. Our preliminary data strongly support the potential of further development towards clinical application in humans. In this proposed project, for clinical relevance, we will test this new scAAV-∆hIDUAop vector in MPS I mouse model using AAV9, via IV, intrathecal (IT) and IV+IT delivery, to assess the therapeutic potential and determine the optimal regimen for treating MPS I. The proposed project will allow us to generate rigorous preclinical efficacy and safety data, to support the subsequent clinical development and commercialization.