Gene Therapy in Hutchinson-Gilford Progeria Syndrome

项目摘要和摘要 Hutchinson-Gilford Progeria综合征(HGPS)是一种无法治愈的、一致致命的疾病，涉及点突变 在一个叫做LMNA(LMNA)的基因中。儿童通常在出生后两年内出现HGPS的迹象， 死亡年龄中值为14岁，最常见的是进行性动脉粥样硬化性心血管疾病。虽然 HGPS的原因突变是在18年前发现的，目前还没有治愈这种疾病的方法。可编程基 编辑DNA现在使以前前所未有的能力能够改变DNA中的单核苷酸并纠正 带有DNA链断裂的致病突变。HGPS代表了一种测试基地编辑的易治疾病， 然而，目前还完全不清楚这种策略是否会改善与以下相关的疾病表型 HGPS。因此，迫切需要研究DNA碱基编辑如何改变驱动HGPS的分子缺陷 以确定这种基因组疗法能否实现其治愈疾病的承诺。我们的总体目标 在这项建议中，发展腺嘌呤碱基编辑(ABE)作为HGPS的治疗策略。我们的中心假设 成年小鼠的ABE治疗可以在动脉中实现足够的编辑，通过 细胞自主对血管平滑肌细胞存活和克隆性增殖的影响。我们的假设是基于 新发表的和新的初步数据表明：1)致病突变的无疤痕矫正 ABE在患者成纤维细胞和人源化HGPS小鼠模型中的应用；2)预防血管病理 Abe处理后6个月幼鼠VSMCs的恢复；3)VSMCs的总体 安倍处理过的幼年动物的存活。该项目基本原理是对基础编辑疗法进行验证 以确定它们在治疗系统性人类疾病方面的潜力。为了实现我们的目标，我们将 追求以下两个具体目标：1)测试DNA编辑是否改善了已建立的 通过一次注射AAV-ABE治疗不同年龄的成年HGPS动物的疾病；2)鉴定 促进ABE治疗后VSMC恢复的机制(S)。这项工作的总体贡献将是 阐明腺嘌呤DNA碱基编辑如何改善HGPS的表型。这项提议的核心创新之处在于 是HGP治疗方法的观念转变，侧重于纠正潜在的致病因素 细胞和组织中的突变。

Project Summary and Abstract Hutchinson-Gilford Progeria Syndrome (HGPS) is an incurable, uniformly fatal disease involving a point mutation in a gene called Lamin A (LMNA). Children develop signs of HGPS typically within the two years after birth and die at a median age of 14, most commonly from progressive atherosclerotic cardiovascular disease. Although the causal mutation in HGPS was identified 18 years ago, no cures for this disease exist. Programmable base editing of DNA now enables the previously unprecedented ability to change single nucleotides in DNA and correct pathogenic mutations with DNA strand breaks. HGPS represents a tractable disease to test base editing, however it remains completely unknown whether this strategy will improve disease phenotypes associated with HGPS. As such, there is a critical need to study how DNA base editing alters the molecular defects driving HGPS in order to determine whether this genome therapy can fulfill its promise to cure disease. Our overall objective in this proposal is develop adenine base editing (ABE) as a treatment strategy for HGPS. Our central hypothesis is that ABE-treatment of adult mice can achieve sufficient editing in aortas to reverse vascular pathology through cell-autonomous effects on survival and clonal proliferation in VSMCs. Our hypothesis is formulated based on newly published and new preliminary data that demonstrate: 1) scarless correction of the pathogenic mutation by ABE in patient fibroblasts and in a humanized mouse model of HGPS; 2) prevention of vascular pathology and recovery of VSMCs at 6 months after ABE treatment of juvenile mice; 3) a significant increase in overall survival of ABE-treated juvenile animals. The rationale for this project is that validation of base editing therapies is needed to determine their potential in treating systemic human diseases. To attain our objectives, we will pursue the following two specific aims: 1) Test whether DNA editing improves vascular pathology in established disease by treating adult HGPS animals at different ages with a single injection of AAV-ABE; 2) Identify the mechanism(s) promoting VSMC recovery after ABE treatment. The overall contribution of this work will be to elucidate how adenine DNA base editing improves phenotypes in HGPS. The central innovation of this proposal is a conceptual shift in therapeutic treatment of HGPs by focusing on correcting the underlying pathogenic mutation in cells and tissues.