Exososomal CNS-Delivery of Therapies for a Lysosomal Disorder

外泌体中枢神经系统治疗溶酶体疾病

• 批准号: 10540037
• 负责人: Gustavo Henrique Boff Maegawa
• 金额: $ 10.21万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-13 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10540037
• 关键词: Exososomal; CNS; Delivery; Therapies; Lysosomal

ABSTRACT Lysosomal storage diseases (LSDs) are genetic disorders caused by pathogenic variants in genes encoding enzymes that are essential for the lysosomal function. These inborn organelle diseases have a cumulative incidence of ~1/2,000-3,000. LSDs have a broad clinical spectrum, mostly associated with a progressive neurodegenerative course, resulting in severe disability and death. The current advances achieved in LSD treatment are exemplified by the enzyme replacement therapy (ERT), which consist of weekly or biweekly intravenous administrations of the recombinant enzymes. Although generally well-tolerated, ERT showed limited biodistribution in the central nervous system (CNS). ERT agents are large molecules, and so they are incapable of crossing the blood-brain barrier (BBB). The lack of reliable and efficacious delivery across the BBB severely limits the ERT and other therapies to treat neurological symptoms. Hence, the development of delivery strategies for therapeutic agents for LSDs is an urgent and unmet need. Unfortunately, 98% of small- molecule drugs and every biological agent, including proteins, cannot cross the BBB. Numerous CNS delivery strategies have been developed to overcome the often stated ‘problem-behind the problem’ of many neurological conditions. Recent studies have shown that exosomes provide a physiological means to deliver proteins or miRNA across BBB. Exosomes, “fingerprints of the cell,” are extracellular nanovesicles are released by cells into the circulation and bodily fluids, displaying distinct cargo profile dependable upon their cellular origin. In 2018, clinic-approved exosomes started to be produced under GMP standards. We hypothesize that exosomes are therapeutic nanovesicles that allow penetration through the BBB to treat neuropathic forms of LSDs. We established a murine neural cell line that secretes neural-derived exosomes containing specific lysosomal enzymes. Leveraging the unique resources we have built over the years, along with our work investigating neurological LSDs and function of exosomes, we will address the following aims: Aim 1. Characterize and further improve the exosomal production from an established murine neural cell line derived from a Twitcher (Twi) mouse model of a severe neurological LSD. We will optimize the production of exosomes from neural immortalized Twi murine cell line stably expressing human GALC (galactocerebrosidase). Using targeted proteomics and sphingolipidomics, we will characterize neuronal exosomes Aim 2. Examine the therapeutic potential of neural-derived exosomes as a CNS- delivery system in the Twitcher, mouse model. We will examine the biodistribution and efficacy of neural- derived exosomes in the neurological course using Twi mice. As mediators of intercellular communication and surrogates of intracellular changes in pathological states, exosomes become a reliable source to deliver therapies to LSDs. This proof-of-principle study will reveal exosomes as a powerful and robust CNS- ‘nanovesicle-delivery’ strategy of therapeutic applications to treat neurodegenerative disorders.

摘要 溶酶体贮藏疾病(LSD)是由编码基因的致病变异引起的遗传病 溶酶体功能所必需的酶。这些先天的细胞器疾病有累积的 发病率为1/2,000-3,000。LSD具有广泛的临床谱系，大多与进行性疾病有关 神经退行性变过程，导致严重残疾和死亡。LSD目前取得的进展 治疗的例子是酶替代疗法(ERT)，它包括每周或两周一次 静脉注射重组酶。虽然总体上耐受性良好，但ERT显示 中枢神经系统(CNS)的生物分布有限。ERT试剂是大分子，它们也是 不能跨越血脑屏障(BBB)。缺乏可靠和有效的跨地区交付 血脑屏障严重限制了ERT和其他治疗神经症状的方法。因此，中国的发展 为LSD提供治疗药物的策略是一个迫切而未得到满足的需求。不幸的是，98%的小型- 分子药物和每一种生物制剂，包括蛋白质，都不能越过血脑屏障。大量CNS交付 已经制定了一些策略来克服许多人经常说的“问题背后的问题” 神经方面的情况。最近的研究表明，外切体提供了一种生理方式来传递 跨血脑屏障的蛋白质或miRNA。胞外体，“细胞的指纹”，是细胞外的纳米囊泡。 由细胞释放到循环和体液中，显示出明显的货物轮廓，取决于它们的 细胞起源。2018年，临床批准的外切体开始根据GMP标准生产。我们 假设外切体是治疗性纳米囊泡，允许通过血脑屏障穿透 治疗神经性形式的LSD。我们建立了一种分泌神经源性物质的小鼠神经细胞系 含有特定溶酶体酶的外切体。利用我们在过去几年中建立的独特资源 几年来，随着我们对神经性LSD和外切体功能的研究，我们将解决 以下目标：目标1.确定并进一步改进已建立的外切体生产 来自严重神经性LSD的Twitcher(TWI)小鼠模型的小鼠神经细胞系。我们 将优化从稳定表达神经永生化的TWI小鼠细胞系中生产外切体 人半乳脑苷酶(GALC)利用靶向蛋白质组学和鞘脂组学，我们将表征 神经源性外切体的目标2.检测神经源性外切体作为中枢神经系统的治疗潜力 在Twitcher小鼠模型中的递送系统。我们将检查神经的生物分布和疗效- 使用TWI小鼠在神经学课程中衍生的外切体。作为细胞间通讯的媒介和 替代细胞内病理状态的变化，外切体成为传递的可靠来源 对LSD的治疗。这项原理验证研究将揭示外体是一个强大而强大的中枢神经系统-- 治疗应用于治疗神经退行性疾病的“纳米胶囊递送”策略。

项目成果

CNS-Targeting Therapies for Lysosomal Storage Diseases: Current Advances and Challenges.

• DOI: 10.3389/fmolb.2020.559804
• 发表时间: 2020
• 期刊: Frontiers in molecular biosciences
• 影响因子: 5
• 作者: Edelmann MJ;Maegawa GHB
• 通讯作者: Maegawa GHB

Antigen-encapsulating host extracellular vesicles derived from Salmonella-infected cells stimulate pathogen-specific Th1-type responses in vivo.

• DOI: 10.1371/journal.ppat.1009465
• 发表时间: 2021-05
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Hui WW;Emerson LE;Clapp B;Sheppe AE;Sharma J;Del Castillo J;Ou M;Maegawa GHB;Hoffman C;Larkin Iii J;Pascual DW;Ferraro MJ
• 通讯作者: Ferraro MJ