Alpha-synuclein degradation mechanisms

α-突触核蛋白降解机制

• 批准号: 8644325
• 负责人: Jianhua Zhang
• 金额: $ 31.09万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-15 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8644325
• 关键词: Affect; Aging; Animal Model; Attenuated; Autophagocytosis; Brain; Cathepsins; Cell model; Cessation of life; Dementia; Dependovirus; Electron Microscopy; Enzymes; Essential Genes; Exhibits; Foundations; Gene Amplification; Gene Mutation; Genetic Transcription; Goals; Immunohistochemistry; In Vitro; Lewy Bodies; Lewy Body Disease; Location; Lysosomes; Mediating; Metabolism; Methods; Molecular; Mus; Mutate; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Organelles; Parkinson Disease; Pathway interactions; Proteins; Regulation; Research; Role; Site; Staining method; Stains; Stress; Substantia nigra structure; Testing; Therapeutic; Toxic effect; Transgenic Mice; Tyrosine 3-Monooxygenase; Up-Regulation; Work; aging brain; alpha synuclein; base; cresyl violet; cytotoxicity; design; dopaminergic neuron; effective therapy; functional decline; gain of function; improved; in vivo; insight; multicatalytic endopeptidase complex; mutant; neuroprotection; neurotoxicity; novel; overexpression; protein aggregate; protein degradation; public health relevance; research study; synucleinopathy; tau Proteins; therapeutic target; treatment strategy

DESCRIPTION (provided by applicant): a-synuclein (a-syn) mutations or gene amplification cause a small subset of Parkinson's disease (PD), with Lewy body (LB) formation, neurodegeneration and often an associated dementia. a-syn aggregation in LB is also widespread in sporadic PD and other LB diseases without apparent upregulation of transcription. a-syn overexpression leads to its aggregation and/or neurotoxicity in various animal models. Reduction of a-syn is neuroprotective in animal models. These observations suggest a therapeutic potential of reducing a-syn in treatment of PD and other LB diseases. Our long-term objective is to determine the mechanisms and regulation of a-synucleinopathy in neurodegenerative diseases, and to provide novel and effective treatment strategies. We focused on the lysosomal function in modulating neuronal a-syn aggregation and toxicity, because lysosomes are high capacity organelles responsible for clearance of damaged and aggregated proteins, and are implicated in aging and several neurodegenerative diseases. We found that mice with deficient lysosomal cathepsin D (CD) exhibited significant a-syn accumulation in the brains, indicating a critical role for CD in mediating a-syn metabolism. In vitro we have shown that overexpression of CD reduces a-syn aggregation and protects against a-syn-mediated toxicity. To further establish CD as a therapeutic target against a-synucleinopathy, we will examine its effects in vivo, and define the mechanisms of its action in vitro. We hypothesize that CD protects against a-syn neurotoxicity by increasing autophagic clearance of toxic species of a-syn. We will test this hypothesis by performing experiments with the following aims: 1. Test the hypothesis that CD haploinsufficiency increases sensitivity to a-syn-induced neurotoxicity in vivo. 2. Test the hypothesis that stereotaxic delivery of AAV-CD to the SN attenuates a-syn-mediated neurotoxicity in vivo. 3. Test the hypothesis that neuroprotection by CD is through clearance of a-syn by autophagy. Completion of these studies will determine the effect of partial loss-of-CD in a-syn-induced DA neuron death, and the effect of gain-of-function of CD in neuroprotection in vivo. Furthermore, insights into the potential molecular mechanisms of CD-mediated neuroprotection and the molecular mechanisms and regulation of clearance of aggregation-prone proteins, will be gained that will permit further refinement of CD therapeutic strategies.

描述（由申请人提供）：a-突触核蛋白（a-syn）突变或基因扩增导致一小部分帕金森病（PD），伴路易体（LB）形成、神经退行性变，通常伴有痴呆。在散发性PD和其他LB疾病中，LB中的a-syn聚集也广泛存在，但没有明显的转录上调。在各种动物模型中，A-syn过表达导致其聚集和/或神经毒性。在动物模型中，a-syn的减少具有神经保护作用。这些观察结果表明，减少a-syn在治疗PD和其他LB疾病方面具有治疗潜力。我们的长期目标是确定a-突触核蛋白病在神经退行性疾病中的机制和调控，并提供新的有效的治疗策略。我们关注的是溶酶体在调节神经元a-syn聚集和毒性方面的功能，因为溶酶体是负责清除受损和聚集蛋白的高容量细胞器，并且与衰老和几种神经退行性疾病有关。我们发现，溶酶体组织蛋白酶D （CD）缺失的小鼠在大脑中表现出显著的a-syn积累，表明CD在介导a-syn代谢中的关键作用。在体外，我们已经证明过表达CD可以减少a-syn聚集，并防止a-syn介导的毒性。为了进一步确定CD作为a-突触核蛋白病的治疗靶点，我们将研究其在体内的作用，并确定其在体外的作用机制。我们假设CD通过增加a-syn有毒物质的自噬清除来保护a-syn神经毒性。我们将通过以下目的的实验来验证这一假设：1。验证CD单倍体功能不全增加体内a-syn诱导的神经毒性敏感性的假设。2. 在体内验证AAV-CD立体定向递送到SN减弱a-syn介导的神经毒性的假设。3. 验证CD的神经保护是通过自噬清除a-syn的假设。这些研究的完成将确定部分CD丢失在a-syn诱导的DA神经元死亡中的作用，以及CD功能获得在体内神经保护中的作用。此外，深入了解CD介导的神经保护的潜在分子机制，以及易于聚集蛋白清除的分子机制和调节，将有助于进一步完善CD治疗策略。

项目成果

Reduction of mutant huntingtin accumulation and toxicity by lysosomal cathepsins D and B in neurons.

• DOI: 10.1186/1750-1326-6-37
• 发表时间: 2011-06-01
• 期刊: Molecular neurodegeneration
• 影响因子: 15.1
• 作者: Liang Q;Ouyang X;Schneider L;Zhang J
• 通讯作者: Zhang J

Inhibition of autophagy and glycolysis by nitric oxide during hypoxia-reoxygenation impairs cellular bioenergetics and promotes cell death in primary neurons.

• DOI: 10.1016/j.freeradbiomed.2013.09.006
• 发表时间: 2013-12
• 期刊: FREE RADICAL BIOLOGY AND MEDICINE
• 影响因子: 7.4
• 作者: Benavides, Gloria A.;Liang, Qiuli;Dodson, Matthew;Darley-Usmar, Victor;Zhang, Jianhua
• 通讯作者: Zhang, Jianhua

Autophagy and Mitophagy in Cellular Damage Control.

• DOI: 10.1016/j.redox.2012.11.008
• 发表时间: 2013
• 期刊: REDOX BIOLOGY
• 影响因子: 11.4
• 作者: Zhang, Jianhua

Bioenergetic and autophagic control by Sirt3 in response to nutrient deprivation in mouse embryonic fibroblasts.

• DOI: 10.1042/bj20130414
• 发表时间: 2013-09-01
• 期刊: The Biochemical journal
• 作者: Liang Q;Benavides GA;Vassilopoulos A;Gius D;Darley-Usmar V;Zhang J
• 通讯作者: Zhang J

Specific distribution of the autophagic protein GABARAPL1/GEC1 in the developing and adult mouse brain and identification of neuronal populations expressing GABARAPL1/GEC1.

自噬蛋白 GABARAPL1/GEC1 在发育中和成年小鼠大脑中的具体分布以及表达 GABARAPL1/GEC1 的神经元群体的鉴定。

• DOI: 10.1371/journal.pone.0063133
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: LeGrand,JaclynNicole;Bon,Karine;Fraichard,Annick;Zhang,Jianhua;Jouvenot,Michèle;Risold,Pierre-Yves;Boyer-Guittaut,Michaël;Delage-Mourroux,Régis
• 通讯作者: Delage-Mourroux,Régis