Development of a Gectosome Therapy for Cardiovascular Diseases

项目摘要 Vesicle Therapeutics Inc旨在开发和商业化一种新的治疗纯合子 家族性高胆固醇血症（hoFH）。大多数hoFH是由两个等位基因的突变引起的 低密度脂蛋白受体（LDL receptor，LDLR）由于他汀类药物和PCSK 9的疗效 抗体治疗在很大程度上取决于功能性LDL受体，hoFH患者表现出有限的 对现有疗法的反应。hoFH没有治愈方法，可用的选择很少 来治疗疾病血管生成素样3（ANGPTL 3）已成为一种可能的治疗方法， hoFH靶点，因为ANGPTL 3缺陷个体不会发生冠状动脉粥样硬化 斑块靶向CRISPR酶LwaCas 13的RNA可以通过RNA耗尽关闭基因 类似于RNAi，但具有非常低的脱靶基因沉默率。安全的缺席 递送方法目前限制了LwaCas 13的治疗潜力。刘实验室在 科罗拉多大学博尔德分校开发了一种创新的细胞内生物制剂输送系统 称为Gectosomes。第一阶段STTR项目的总体目标是证明， 通过LwaCas 13 a/ANGPTL 3 crRNA的外泌体递送来沉默ANGPTL 3有效地抑制ANGPTL 3的表达。 降低LDL-C，在小鼠中具有可接受的安全性特征。拟议战略结合了两个 创新技术，用于潜在的临床翻译。建议的研究是可行的 我们之前成功地用CRISPR RNP的外泌体递送， 小鼠肝脏中的PCSK 9。ANGPTL 3主要在肝细胞中表达，并分泌到肝细胞中。 血流肝易于被外泌体接近。LwaCas 13 a介导的RNA缺失 是可逆的，并且可能比基因编辑具有更少的安全性问题。康贝特人将以 LwaCas 13 a在小鼠中抑制ANGPTL 3的有效性和安全性，这项工作是必要的 进一步研究，以推进一种罕见疾病的潜在治疗方案。

Project Summary Vesicle Therapeutics Inc aims to develop and commercialize a new therapy for homozygous familial hypercholesterolemia (hoFH). A majority of hoFH is caused by mutations in both alleles of the gene encoding the LDL receptor (LDLR). Since the efficacy of both statins and PCSK9 antibody therapies largely depends on functional LDL receptors, patients with hoFH show limited responses to these existing therapies. There is no cure for hoFH, and few options are available to treat the diseases. Angiopoietin-like 3 (ANGPTL3) has emerged as a possible therapeutic target for hoFH as individuals deficient in ANGPTL3 do not develop coronary atherosclerotic plaque. The RNA targeting CRISPR enzyme LwaCas13 can turn off genes by RNA depletion analogous to RNAi but with very lower rate of off-target gene silencing. The absence of safe delivery methods currently limits the therapeutic potential of LwaCas13. The Liu laboratory at the University of Colorado-Boulder developed an innovative intracellular biologics delivery system called Gectosomes. The overall objective of this phase I STTR project is to demonstrate that silencing of ANGPTL3 by gectosome delivery of LwaCas13a/ANGPTL3 crRNA is efficacious in lowering LDL-C with acceptable safety profile in mice. The proposed strategy combines two innovative technologies for potential clinical translation. The proposed studies are feasible based on our previous success with gectosome delivery of CRISPR RNP that causes inactivation of PCSK9 in mouse liver. ANGPTL3 is primarily expressed in hepatocytes and secreted into the bloodstream. The liver is readily accessible by gectosomes. LwaCas13a-mediated RNA depletion is reversible and may have fewer safety concerns than gene editing. We will determine the efficacy and safety of ANGPTL3 suppression by LwaCas13a in mice and this work is necessary for further studies to advance a potential therapeutic solution for a rare disease.