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 溶酶体贮积症（LSD）是一组以功能障碍为特征的遗传性疾病 溶酶体，累积频率为七千分之一的活产中，超过 2/3 的 LSD 患者出现受累。 中枢神经系统 (CNS) 的严重程度广泛 (nLSD)，这使得 LSD 成为最严重的 儿童神经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，但它们大多是。 由于酶对中枢神经系统的渗透性较差，未能成功逆转神经系统并发症， 治疗 nLSD 的一个主要障碍 拟议研究的影响是由未满足的医疗需求驱动的。 遗传性 nLSD 的有效治疗以及酶输送到 CNS 的主要限制。 阳离子非依赖性甘露糖 6 磷酸受体 (M6PR) 在溶酶体酶中发挥关键作用 大多数溶酶体酶的运输和细胞间转移，这对于代谢交叉至关重要 纠正产后早期 M6PR 对血脑屏障 (BBB) 的发育下降。 小鼠和人类的这一时期归因于缺乏将中枢神经系统酶输送到成人大脑中。 具有位点突变的荧光素酶报告系统，我们最近将 microRNA-143 (miR143) 鉴定为 使用小鼠模型降低脑微血管 (BrMV) 上的 M6PR 蛋白水平的表观遗传调节剂。 Hurler 综合征（I 型严重粘多糖贮积症，MPS I），由 α-L- 缺乏引起 艾杜糖醛酸酶 (IDUA)，我们证明了 M6PR 介导的 IDUA 在双脑大脑中转移的功能性拯救 敲除 (MPS/miR-143KO) 小鼠具有长期中枢神经系统治疗益处，以及对人类血管的益处 通过将 miR-143 与 miR-143-海绵序列隔离来观察内皮细胞的结果提供了强有力的数据。 开发一种新方法的科学前提，该方法将有选择地向系统性“开放”BBB 当前任何治疗方案或未来酶/基因/细胞疗法为协同中枢神经系统提供的酶 在这项提案中，我们的目标是开发一种基于腺相关病毒载体（AAV）的药物。 可翻译平台“恢复”成熟 BBB 上的 M6PR 通路，以先进地递送治疗酶 进入中枢神经系统有 3 个目标，包括开发人工 miR143 抑制剂 (143in) 和表达 用于强力和有针对性地减少脑内皮细胞上 miR143 的盒（目标 1），体内检查 AAV/143in 递送小鼠的“中靶”和“脱靶”表达和效果（目标 2）以及临床前 BrMV 靶向 AAV/143in 通过酶疗法纠正 MPS I 小鼠中枢神经系统异常的评估 源自转基因红细胞/巨核细胞谱系（目标 3）。 一种新的体内 miRNA 抑制剂介导的脑靶向方法的概念，该方法可适用于 许多其他涉及 M6PR 通路的 nLSD 和神经系统疾病受益于先进的 CNS 传递 通过用 M6P 残基或 IGF2 标签进行修饰来调整 M6PR 介导的转运途径来进行治疗。