Exosome based therapeutics in Huntington's disease

基于外泌体的亨廷顿病疗法

基本信息

批准号：
8581918
负责人：
NEIL ARONIN
金额：
$ 50万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-01 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8581918
关键词：
Adolescence Adolescent Adult Affect Age Age-Years Alleles Animal Model Antigens Antisense Oligonucleotides Area Behavior Behavior assessment Blood Blood - brain barrier anatomy Blood Circulation Brain Brain Diseases 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 Immune Immune response Impaired cognition Infusion procedures Inherited Injection of therapeutic agent Measurement Measures Memory Mental Depression Messenger RNA Methods MicroRNAs Movement Multivesicular Body Mus Nerve Degeneration Neuroglia Neurons Parents Pathogenesis Pathway interactions Patients 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 Time Trinucleotide Repeats Variant Veins base brain cell combinatorial deep sequencing human DICER1 protein human Huntingtin protein immunogenicity in vivo knock-down mouse model mutant neuropathology nonhuman primate prevent public health relevance rabies virus glycoprotein G small hairpin RNA theories uptake

项目摘要

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跨血脑屏障传递 siRNA 进入神经元和其他脑细胞，直接目标是突变型亨廷顿病 (HD)。由 CAG 三核苷酸重复次数增加引起 ¿系列 36；由于纹状体和皮质神经退行性变，HD 患者会出现认知障碍、抑郁和运动异常，因此需要在高级护理机构中呆上数年。少数患者在青少年时期发病，合理的治疗方法是减少亨廷顿蛋白突变体 mRNA 的表达；这种治疗方法可以通过 siRNA、反义寡核苷酸 (ASO) 和腺相关药物在 HD 小鼠模型中实现。然而，在非人类灵长类动物中，递送 siRNA 和 ASO 并不能到达纹状体和纹状体。尽管 siRNA 在大脑中的传播受到限制，但 AAV-shRNA 需要多次注射到大脑区域，而 HD 治疗中的空白可以通过通常挤出的微泡来填补。通过细胞，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，并进入神经元和神经胶质细胞，RVG-外泌体可以携带siRNA货物进入血液循环。外泌体将 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 的治疗剂。将研究针对亨廷顿蛋白 mRNA 穿过血脑屏障进入神经元的 siRNA 定位和 RNAi 依赖性敲低，因为外泌体是由细胞质、外泌体 mRNA、miRNA 和植入的。 siRNA 将通过深度测序进行鉴定，并且将研究免疫反应性和免疫中和作用，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 小鼠模型中的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。外泌体生产和脑递送将进行研究，利用外泌体进行脑递送有望形成一种可行的治疗方法，以减少患有其他遗传性疾病的患者中突变亨廷顿蛋白的表达。神经退行性疾病将会受益。