Validating cGAS-STING pathway as drug target in Huntington disease mouse model

在亨廷顿病小鼠模型中验证 cGAS-STING 通路作为药物靶点

基本信息

批准号：
10508092
负责人：
Srinivasa Subramaniam
金额：
$ 53.13万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-20 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10508092
关键词：
Address Affect Amyotrophic Lateral Sclerosis Anatomy Animal Diseases Anti-Inflammatory Agents Atrophic Autophagocytosis Behavioral Body Weights and Measures CAG repeat Catabolic Process Cell Culture Techniques Cell model Cells Clinical Trials Code Corpus Callosum Corpus striatum structure Cyclic GMP Data Defect Development Disease Disease Progression Disease model Drug Targeting Etiology Exons Functional disorder Gene Activation Genes Genetic Genetic Transcription Human Huntington Disease Huntington gene Immunohistochemistry Impaired cognition Inflammation Inflammatory Inflammatory Response Knock-in Mouse Knock-out Knockout Mice Knowledge Kynurenine Link Mediating Messenger RNA Microglia Mission Modeling Modification Molecular Monkey Diseases Motor Motor Activity Mus Mutation Natural Immunity Nerve Degeneration Neurodegenerative Disorders Onset of illness Pain Pathogenesis Pathogenicity Pathologic Pathology Pathway interactions Patients Performance Peripheral Persons Pharmaceutical Preparations Pharmacology Phenotype Pre-Clinical Model Public Health Research Research Personnel Ribosomes Role Severity of illness Signal Pathway Signal Transduction Stimulator of Interferon Genes Symptoms Techniques Testing Therapeutic Therapeutic Agents Time Tissue Model Tissues Tryptophan United States National Institutes of Health Walking Western Blotting Work age related base brain tissue chemokine cytokine disability disease phenotype effective therapy experimental study inhibitor insight intraperitoneal monocyte mouse model mutant nervous system disorder novel novel therapeutic intervention polyglutamine pre-clinical prevent response small molecule small molecule inhibitor targeted treatment therapeutic target transcription factor transcriptome sequencing

项目摘要

Project summary Huntington disease (HD) is a fatal neurodegenerative disorder caused by the huntingtin (HTT) CAG expansion mutation (coding for polyglutamine, mHTT). mHTT is a ubiquitously expressed gene, yet it prominently damages the striatum and cortex, followed by widespread peripheral defects as the disease progresses. Prior works have linked innate and adaptive inflammatory responses in HD. Microglia, a cellular indicator of inflammation, is also increased in the striatum of HD animal and cell culture models and HD patients. Increased levels of reactive monocytes, inflammatory cytokines, chemokines, and the n-kynurenine/tryptophan ratio, an indicator of persistent inflammation, have all been observed in pre-manifest HD patients and correlated with HD progression. Furthermore, RNA-seq analysis of tissue obtained from human HD patients and HD monkey models reveals extensive transcriptional dysregulation associated with proinflammatory pathway activation. Inflammation is also closely linked to autophagy, a catabolic process that is dysregulated in HD. Despite these studies, the mechanisms or treatment targets for the reduction of inflammatory responses remain less clear. To fill this knowledge gap, this proposal tests the hypothesis that cyclic GMP–AMP synthase (cGAS)–stimulator of interferon genes (STING), a major innate immunity response pathway, is a disease modifier in HD pathogenesis. This hypothesis was formulated based on our recent finding that the cGAS-STING pathway is upregulated in HD patients’ tissue and cell models and mediates autophagy and inflammatory responses. We found high ribosome occupancy in exon 1 of cGAS mRNA and cGAS-dependent inflammatory transcription factors (Irf3, Irf7), and inflammatory chemokine (Ccl5, and Cxcl10) are upregulated in HD. Depletion of cGAS diminishes downstream effector STING activation and also inhibits autophagy flux in HD cells. The latter is consistent with the primordial function of the cGAS-STING pathway in autophagy induction. These data underline an important role of cGAS-STING signaling in the pathological mechanisms of HD. Subsequently, an independent study confirmed activation of cGAS-STING in HD models. Yet, it is unclear whether the cGAS- STING pathway is a passive or active contributor to HD development. In Aim 1, we will determine whether the genetic deletion of cGAS alters the behavioral and pathological hallmarks of HD KI (z175HD) mice. We will determine age-related behavioral alterations and pathological changes in cGAS–/–;HD KI mice. In Aim 2, we will determine whether the small molecule STING inhibitor diminishes the HD phenotype in HD KI mice. We will treat z175HD mice with the small molecule STING inhibitor, H-151 and determine changes in the HD-like deficits compared vehicle treated z175HD. The proposed experiments will clarify the role of the cGAS–STING pathway as anti-inflammatory therapeutic agents in HD.

项目摘要亨廷顿病（HD）是由亨廷顿（HTT）CAG扩张引起的致命神经退行性疾病突变（编码聚谷氨酰胺，MHTT）。 MHTT是一个普遍表达的基因，但它显着损坏纹状体和皮质，随着疾病进展的宽度宽度。先前的作品有 HD中有联系的先天和适应性炎症反应。小胶质细胞是炎症的细胞指标，也是 HD动物和细胞培养模型和HD患者的纹状体中的增加。反应性水平增加单核细胞，炎性细胞因子，趋化因子和N-酮氨酸/色氨酸比率，这是一个指标持续性炎症，全部观察到了手势前HD患者，并且与HD进展相关。此外，从人类高清患者和HD猴子模型获得的组织的RNA-seq分析显示与促炎途径激活相关的广泛转录失调。炎症也是与自噬密切相关，自噬是一种在HD中失调的分解代谢过程。尽管进行了这些研究，减少炎症反应的机制或治疗目标尚不清楚。填写这个知识差距，该建议检验了环环GMP – AMP合酶（CGA）的假设 - 干扰素基因（Sting）是一种主要的先天免疫反应途径，是HD中的疾病修饰剂发病。该假设是根据我们最近的发现，即CGAS刺道途径是在HD患者的组织和细胞模型中进行了更新，并介导自噬和炎症反应。我们发现在CGAS mRNA和CGAS依赖性炎症转录的外显子1中发现高核糖体占用率因素（IRF3，IRF7）和炎症趋化因子（CCL5和CXCL10）以HD更新。 CGA的耗竭减少下游效应子的激活，并抑制HD细胞中的自噬通量。后者是与自噬诱导中CGAS刺途径的原始功能一致。这些数据强调 CGAS信号传导在HD的病理机制中的重要作用。随后，一个独立研究证实了HD模型中CGAS刺的激活。但是，尚不清楚CGAS-是否 Sting Pathway是高清开发的被动或积极贡献者。在AIM 1中，我们将确定是否 CGA的遗传缺失改变了HD KI（Z175HD）小鼠的行为和病理标志。我们将确定与年龄相关的行为改变和CGA中的病理变化 - / - ； HD KI小鼠。在AIM 2中，我们将确定小分子刺激抑制剂是否会降低HD Ki小鼠的HD表型。我们将对待 Z175HD小鼠具有小分子刺激器H-151并确定HD样缺陷的变化比较了用车辆处理的Z175HD。提出的实验将阐明CGA -sting途径的作用作为HD中的抗炎治疗剂。