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(申请人提供)：UH2和UH3的目标是研究外体如何通过血脑屏障运送siRNAs进入神经元和其他脑细胞。直接的目标是突变的Huntingtin mRNA。亨廷顿病(HD)是由CAG三核苷酸重复序列增加到�36引起的；由于神经退行性变，首先是纹状体和皮质，然后是其他大脑结构，因此需要在高水平的护理设施中呆上数年。HD患者有认知障碍、抑郁和运动异常。大多数HD患者的年龄在30-40岁之间，少数人起病为青少年。一种合理的治疗方法是减少突变的Huntingtin mRNA的表达；在HD小鼠模型中，这种治疗可以通过siRNA、反义寡核苷酸(ASO)和腺相关病毒(AAV)与针对Huntingtin mRNA的shRNAmir来完成。然而，分娩仍然是这些疗法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，将ASO注射到脊髓液中不会到达纹状体，siRNA在大脑中的传播也受到限制。尽管前景看好，AAV-shRNA需要向大脑区域注射几次，而shRNAmir不受监管。HD疗法中的空白可以由通常由细胞、外切体挤出的微囊来填补。表面带有狂犬病病毒糖蛋白(RVG)的外切体可以被注射到血液中，穿过血脑屏障，进入神经元和胶质细胞。RVG-exosome可以携带siRNA货物。外切体进入血液循环，在神经元中沉积siRNA，参与RNA干扰。我们的目的是开发外切体作为HD的治疗药物。UH2检测携带针对Huntingtin mRNA的siRNA的RVG-exosome跨越血脑屏障进入神经元的能力。脑内定位和依赖RNAi的基因敲除将被研究。人们将寻找功能超强的siRNA。由于外体是由细胞质组成的，因此外体mRNA、miRNA和嵌入的siRNA将通过深度测序来鉴定。将研究免疫反应性和免疫中和，因为外切体具有潜在的抗原，如RVG，需要经常给药。UH3通过逆转或预防HD小鼠模型的神经病理和异常运动的研究，进一步建立了基于外切体的治疗方法。外切体的剂量将得到保障。一个由HD发病机制、siRNA开发、RNA鉴定和测量、RNAi机制以及外切体生产和脑递送方面的专家团队将开展这些研究。利用外切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变性的患者将受益。