Exosome based therapeutics in Huntington's disease

基于外泌体的亨廷顿病疗法

• 批准号: 9325094
• 负责人: NEIL ARONIN
• 金额: $ 97.24万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9325094
• 关键词: Adolescence; Adolescent; Adult; Affect; Age; Age-Years; Alleles; Animal Disease Models; Antigens; Antisense Oligonucleotides; Area; Behavior; Behavior assessment; Blood; Blood - brain barrier anatomy; Blood Circulation; Brain; Brain Diseases; CAG repeat; Caring; Catheters; Cells; Cerebrospinal Fluid; Child; Cleaved cell; Clinical; Cognitive; Corpus striatum structure; Cytoplasm; Dependovirus; Deposition; Development; Discipline of Nursing; Disease; Dose; Dyskinetic syndrome; Foundations; Gene Silencing; Genetic; Goals; Human; Huntington Disease; Huntington gene; Immune; Immune response; Impaired cognition; Infusion procedures; Inherited; Injectable; Injection of therapeutic agent; Measurement; Measures; Memory; Mental Depression; Messenger RNA; Methods; MicroRNAs; Movement; Multivesicular Body; Mus; Nerve Degeneration; Neuroglia; Neurons; Nurses; Parents; Pathogenesis; Pathway interactions; Patients; Peripheral; Prevention; Production; Proteins; RNA; RNA Interference; RNA Splicing; Risk; Secure; Series; Site; Small Interfering RNA; Small RNA; Source; Structure; Surface; Tail; Testing; Therapeutic; Therapeutic Studies; Treatment Efficacy; Variant; Veins; base; brain cell; combinatorial; cost; deep sequencing; exosome; gene product; immunogenicity; in vivo; knock-down; microvesicles; mouse model; mutant; neuropathology; nonhuman primate; prevent; public health relevance; rabies virus glycoprotein G; small hairpin RNA; theories; uptake

DESCRIPTION (provided by applicant): The goal of this UH2 and UH3 is to study how exosomes can deliver siRNAs across the blood brain barrier to enter neurons and other brain cells. The immediate target is the mutant huntingtin mRNA. Huntington's disease (HD) is caused by an increase in the CAG trinucleotide repeats to � 36 in series; it necessitates years in a high level nursing facility because of neurodegeneration first in striatum and cortex and then to other brain structures. HD patients have cognitive impairment, depression and aberrant movements. Most HD patient present by 30 to 40 years of age; a few have a juvenile onset. A rational treatment is to decrease expression of mutant huntingtin mRNA; this therapeutic can be accomplished in HD mouse models by siRNA, antisense oligonucleotides (ASO) and adeno-associated virus (AAV) with shRNAmir directed against huntingtin mRNA. However, delivery remains a pitfall to practical implementation of the therapeutics. siRNA and ASO require long-term infusion. In non-human primates, ASO administered to spinal fluid does not reach the striatum and spread of siRNA is limited in brain. Although promising, AAV-shRNA requires several injections into brain areas and the shRNAmir is unregulated. A gap in HD therapeutics can be filled by microvesicles normally extruded by cells, exosomes. Exosomes with rabies virus glycoprotein (RVG) on their surface can be injected into the blood, cross the blood brain barrier, and enter neurons and glia. RVG-exosomes can carry siRNA cargo. Delivered into the blood circulation, the exosomes deposit siRNA in neurons to engage in RNA interference. Our purpose is to develop exosomes as a therapeutic in HD. The UH2 examines the ability of RVG-exosomes carrying siRNA against huntingtin mRNA to cross the blood brain barrier to enter neurons. Localization in brain and RNAi dependent knock down will be studied. Hyper-functional siRNAs will be sought. Because exosomes are made from cytoplasm of cells, exosome mRNA, miRNA, and implaced siRNA will be identified by deep sequencing. Immune reactivity and immune-neutralization will be studied, since exosomes have potential antigens, like RVG, and will need to be administrated often. The UH3 further establishes exosome-based therapeutics, by study of reversal or prevention of neuropathology and aberrant movement in HD mouse models. Dosing of exosomes will be secured. A team of experts in HD pathogenesis, siRNA development, RNA identification and measurement, RNAi mechanisms and exosome production and brain delivery will carry out the studies. Harnessing exosomes for brain delivery is expected to form a viable therapeutic to reduce expression of mutant huntingtin in patients with HD. Patients with other genetically- based neurodegeneration will benefit.

描述（由申请人提供）：该 UH2 和 UH3 的目标是研究外泌体如何递送 siRNA 穿过血脑屏障进入神经元和其他脑细胞。直接目标是突变亨廷顿蛋白 mRNA。亨廷顿舞蹈病 (HD) 是由 CAG 三核苷酸重复序列增加至 36 个引起的；由于纹状体和皮质首先发生神经退行性变，然后是其他大脑结构，因此需要在高级护理机构中呆上数年。 HD 患者存在认知障碍、抑郁和运动异常。大多数 HD 患者发病年龄为 30 至 40 岁；少数青少年发病。合理的治疗方法是减少突变型亨廷顿蛋白 mRNA 的表达；这种治疗方法可以在 HD 小鼠模型中通过 siRNA、反义寡核苷酸 (ASO) 和腺相关病毒 (AAV) 以及针对亨廷顿蛋白 mRNA 的 shRNAmir 来实现。然而，递送仍然是治疗实际实施的一个陷阱。 siRNA和ASO需要长期输注。在非人类灵长类动物中，给予脊髓液的 ASO 不会到达纹状体，并且 siRNA 在大脑中的传播受到限制。尽管前景广阔，但 AAV-shRNA 需要多次注射到大脑区域，而且 shRNAmir 不受监管。 HD 疗法的空白可以通过通常由细胞挤出的微泡、外泌体来填补。表面带有狂犬病病毒糖蛋白（RVG）的外泌体可以被注射到血液中，穿过血脑屏障，进入神经元和神经胶质细胞。 RVG-外泌体可以携带 siRNA 货物。外泌体被输送到血液循环中，将 siRNA 沉积在神经元中以参与 RNA 干扰。我们的目的是开发外泌体作为 HD 的治疗方法。 UH2 检查携带针对亨廷顿蛋白 mRNA 的 siRNA 的 RVG 外泌体穿过血脑屏障进入神经元的能力。将研究大脑中的定位和 RNAi 依赖性敲除。人们将寻求超功能的 siRNA。由于外泌体是由细胞的细胞质制成的，因此外泌体的 mRNA、miRNA 和植入的 siRNA 将通过深度测序进行鉴定。将研究免疫反应性和免疫中和，因为外泌体具有潜在的抗原，如 RVG，并且需要经常施用。 UH3 通过研究 HD 小鼠模型中神经病理学和异常运动的逆转或预防，进一步建立了基于外泌体的疗法。外泌体的剂量将得到保证。 HD发病机制、siRNA开发、RNA鉴定和测量、RNAi机制以及外泌体产生和脑递送方面的专家团队将开展这些研究。利用外泌体进行脑递送有望形成一种可行的治疗方法，以减少 HD 患者突变亨廷顿蛋白的表达。患有其他遗传性神经退行性疾病的患者将会受益。