Advancing CNS drug delivery via epigenetic modulation

通过表观遗传调节促进中枢神经系统药物输送

• 批准号: 10679755
• 负责人: Dao Pan
• 金额: $ 58.37万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-18 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679755
• 关键词: Adult; Aftercare; Allogenic; Behavioral; Blood - brain barrier anatomy; Brain; Capsid; Cell Therapy; Cells; Childhood; Circulation; DNA cassette; Data; Dependovirus; Development; Disease; Down-Regulation; Drug Delivery Systems; Enzymes; Epigenetic Process; Erythroid; Excision; Foundations; Frequencies; Functional disorder; Future; Gene Transfer; Genes; Genetic; Glycosaminoglycans; Goals; Hematopoietic Stem Cell Transplantation; Hereditary Disease; Histopathology; Human; Hybrids; IGF Type 2 Receptor; IGF2 gene; IGF2R gene; Inherited; Knock-out; L-Iduronidase; Live Birth; Luciferases; Lysosomal Storage Diseases; Lysosomes; MEKs; Mediating; Medical; Megakaryocytes; Messenger RNA; Metabolic; MicroRNAs; Modeling; Modification; Mucopolysaccharidosis I; Mucopolysaccharidosis I H; Mus; Mutagenesis; Nerve; Neurodegenerative Disorders; Neurologic; Organ; Pathologic; Pathway interactions; Patients; Penetration; Peripheral; Play; Porifera; Production; Receptor Down-Regulation; Reporter; Resolution; Role; Severities; Site; Specificity; System; Testing; Therapeutic; Transfection; Translations; Up-Regulation; Variant; Vascular Endothelial Cell; Viral Vector; Visceral; adeno-associated viral vector; brain cell; brain endothelial cell; caspase 14; cell type; cerebral microvasculature; correctional system; derepression; design; enzyme replacement therapy; enzyme therapy; improved; in vivo; inhibitor; intravenous injection; macromolecule; microRNA delivery; mouse model; nervous system disorder; novel; novel strategies; postnatal period; preclinical evaluation; promoter; prototype; receptor; receptor downregulation; receptor expression; therapeutic enzyme; therapeutic protein; trafficking; transgene expression

Abstract Lysosomal storage disorders (LSDs) are a group of inherited diseases characterized by dysfunctions in lysosomes, with cumulative frequency of 1 in 7000 live births. Over 2/3 of LSD patients present an involvement of the central nerve system (CNS) with a broad spectrum of severity (nLSD), which makes LSDs the most common cause of pediatric neuronopathic diseases. Allogeneic hematopoietic stem cell transplantation (HSCT) or enzyme replacement therapy (ERT) are main treatment options for LSDs. However, they are largely unsuccessful in reversing neurological complications due to the poor penetration of the enzymes into the CNS, a major obstacle in treating nLSD. The impact of the proposed study is driven by the unmet medical need for efficient treatment of inherited nLSDs AND the major limitation of enzyme-delivery into the CNS. The cation-independent mannose-6-phosphate receptor (M6PR) plays a critical role in lysosomal enzyme trafficking and intercellular transfer for the majority of lysosomal enzymes, which is essential for metabolic cross- correction in treating LSDs. Developmental decline of M6PR on blood-brain-barrier (BBB) during early postnatal period in mouse and human is attributable to the lack of CNS enzyme delivery into adult brain. Using a dual luciferase reporter system with site-mutagenesis, we have recently identified microRNA-143 (miR143) as an epigenetic modulator to reduce M6PR protein levels on brain microvessels (BrMV). Using a mouse model of Hurler syndrome (severe mucopolysaccharidosis type I, MPS I), which is caused by the deficiency of α-L- iduronidase (IDUA), we demonstrated functional rescue of M6PR-mediated IDUA transfer in the brain of double- knockout (MPS/miR-143KO) mice with long-term CNS therapeutic benefits, as well as in human vascular endothelial cells by sequestration of miR-143 with miR-143-sponge sequences. The data provide strong scientific premise for the development of a novel approach that would selectively “open” BBB to systemic enzymes provided by any current treatment options or future enzyme/gene/cell therapies for synergistic CNS benefits in many nLSDs. In this proposal, we aim to develop an adeno-associated viral vector (AAV)-based translatable platform to “restore” M6PR pathway on mature BBB for advanced delivery of therapeutic enzymes into the CNS with 3 aims, including developing optimal artificial miR143 inhibitor (143in) and expression cassette(s) for robust and targeted reduction of miR143 on brain endothelia cells (Aim 1), in vivo examination of “on-target” and “off-target” expression and effects in mice with AAV/143in delivery (Aim 2), as well as preclinical evaluation of BrMV-targeted AAV/143in in correcting CNS abnormalities in MPS I mice by enzyme therapy derived from genetically modified erythroid/megakaryocytic lineages (Aim 3). The studies will provide a proof- of concept for a new in vivo miRNA-inhibitor mediated, brain-targeted approach that could be applicable for many other nLSDs involving M6PR pathway AND neurological diseases benefiting from advanced CNS delivery of therapeutics via adapting M6PR-mediated transport pathway by modification with M6P residues or IGF2-tag.

抽象的 溶酶体存储障碍（LSD）是一组遗传性疾病 溶酶体，累积频率为7000个活生生中的1个。超过2/3的LSD患者参与 具有广泛严重程度（NLSD）的中枢神经系统（CNS），这使LSD成为最多 小儿神经疾病疾病的常见原因。同种异体造血干细胞移植（HSCT） 或酶替代疗法（ERT）是LSD的主要治疗选择。但是，他们在很大程度上是 由于酶进入中枢神经系统的渗透率不佳，无法逆转神经系统并发症 治疗NLSD的主要障碍。拟议的研究的影响是由未满足的医疗需求驱动的 有效治疗遗传的NLSD和酶 - 递送到中枢神经系统的主要局限性。 非阳离子甘露糖-6-磷酸受体（M6PR）在溶酶体酶中起关键作用 大多数溶酶体酶的运输和细胞间转移，这对于代谢跨度至关重要 治疗LSD的校正。 M6PR在产后早期血脑屏障（BBB）上的发育下降 小鼠和人的周期归因于缺乏中枢神经系统酶递送到成年大脑中。使用双重 荧光素酶记者系统具有现场 - 毒素，我们最近将MicroRNA-143（miR143）确定为 表观遗传调节剂可降低脑微血管上的M6PR蛋白水平（BRMV）。使用鼠标模型 班勒综合征（严重的粘多糖含量I型，MPS I），这是由α-L-缺乏引起的 iDuronidase（idua），我们证明了在双重介导的M6PR介导的IDUA转移的功能性拯救 具有长期CNS治疗益处的敲除（MPS/mir-143KO）小鼠以及人血管 通过miR-143与miR-143海绵序列的miR-143序列的内皮细胞。数据提供强大 开发一种新型方法的科学前提，该方法将有选择地“开放” BBB到系统 任何当前治疗方案或未来酶/基因/细胞疗法提供的酶为协同中枢神经系统 许多NLSD的好处。在此提案中，我们旨在开发基于腺相关的病毒载体（AAV） 可翻译的平台，可在成熟BBB上“还原” M6PR途径，用于先进的治疗酶 带有3个目标的中枢神经系统，包括开发最佳人工mir143抑制剂（143英寸）和表达 磁带（S）用于在脑内皮细胞上稳健和靶向降低miR143（AIM 1），体内检查 AAV/143英寸递送（AIM 2）以及临床前的小鼠中的“靶标”和“脱靶”表达和效果 通过酶治疗纠正MPS I小鼠中枢神经系统异常的评估，对BRMV靶向的AAV/143IN评估 源自一般修饰的红细胞/巨核细胞谱系（AIM 3）。研究将提供证明 - 一种新的体内miRNA抑制剂介导的，脑针对性的方法的概念，可以适用 许多其他涉及M6PR途径和神经系统疾病的NLSD受益于高级中枢神经系统的交付 通过用M6P残差或IGF2-TAG修饰M6PR介导的传输途径的理论。