Gene therapy for disorders of the extracellular matrix

项目摘要 COL 4A 1和COL 4A 2突变导致Gould综合征（GS）-一种多系统疾病， 异质性脑血管病是主要后果。脑血管病患者 GS的范围可以从由子宫内的生殖器基质损伤引起的脑穿通症，到婴儿癫痫发作，到老年痴呆症。 相关的脑小血管病（cSVD）和血管性认知障碍和痴呆（VCID）。标志 在GS患者中观察到的cSVD包括皮质下微出血，血管周围间隙扩大， 腔隙性梗塞重要的是，Col 4a 1突变小鼠忠实地模拟患者表型。此外，Col 4a 1突变体 小鼠具有与年龄相关的脑血管功能障碍，包括肌原性张力丧失和充血性心力衰竭受损。 这些反应被认为对VCID进展至关重要。 由COL 4A 1和COL 4A 2引起的细胞外损伤是异质的和复杂的，这代表了 这是基于机制的干预措施的一个重大障碍。然而，由于GS是一种毁灭性的单基因 疾病与一个明确的遗传原因，它是一个理想的候选人，纠正疾病的根本原因，通过 基因组编辑技术。在这项提案中，我们将利用大大改进的CRISPR核酸酶，碱基编辑器， 和引物编辑器沿着与新的病毒载体一起测试使用原代GS患者细胞的治疗方法 和GS的小鼠模型。该项目将为开发第一个基因组编辑提供重要的临床前数据- 治疗这种严重的单基因疾病这项工作的成功完成最终可以 为GS患者提供预防儿童卒中和年龄相关VCID的一次性终身治疗 并为类似疾病的纠正制定路线图。

PROJECT SUMMARY COL4A1 and COL4A2 mutations cause Gould syndrome (GS) – a multisystem disorder for which clinically heterogeneous cerebrovascular disease is the major consequence. Cerebrovascular disease in individuals with GS can range from porencephaly caused by germinal matrix hemorrhages in utero, to infantile seizures, to age- related cerebral small vessel disease (cSVD) and vascular cognitive impairment and dementia (VCID). Hallmarks of cSVD observed in individuals with GS include subcortical microbleeds, enlarged perivascular spaces, and lacunar infarcts. Importantly, Col4a1 mutant mice faithfully model patient phenotypes. Moreover, Col4a1 mutant mice have age-related cerebrovascular dysfunction including loss of myogenic tone and impaired hyperemic responses that are thought to be critical to VCID progression. The extracellular insults resulting from COL4A1 and COL4A2 are heterogeneous and complex, which represents a significant barrier to mechanism-based interventions. However, because GS is a devastating monogenic disease with a defined genetic cause, it is an ideal candidate for correction of the root cause of the disease via genome editing technologies. In this proposal, we will leverage vastly improved CRISPR nucleases, base editors, and prime editors along with novel viral vectors to test therapeutic approaches using primary GS patient cells and mouse models of GS. This project will provide important pre-clinical data to develop the first genome editing- based therapy for this severe monogenic disorder. The successful completion of this work could eventually provide a one-time, lifelong treatment that prevents both childhood stroke and age-related VCID for GS patients and create a roadmap for correction of similar diseases.