SOD1/Bcl-2 induced mitochondrial dysfunction in FALS and SALS.

SOD1/Bcl-2 诱导 FALS 和 SALS 线粒体功能障碍。

基本信息

批准号：
8600732
负责人：
PIERA PASINELLI
金额：
$ 34万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-08-02 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8600732
关键词：
Affect Amyotrophic Lateral Sclerosis Astrocytes BCL2 gene BH3 Domain Binding Cell Death Cells Cessation of life Coculture Techniques Complex Data Defect Development Diagnosis Disease Etiology Exposure to Family Functional disorder Gene Mutation Genetic Goals Impairment In Vitro Knock-in Mouse Link Mediating Metabolic Microglia Minority Mitochondria Morphology Motor Neurons Mus Mutate Nerve Degeneration Neurodegenerative Disorders Neuroglia Onset of illness Paralysed Pathogenesis Pathology Pathway interactions Patients Peptides Proteins Reporting Role Scientist Specificity Spinal Cord System Testing Therapeutic Effect Toxic effect Toxicant exposure Transgenic Mice Transgenic Organisms Treatment Efficacy Voltage-Dependent Anion Channel base design immortalized cell in vivo lymphoblast mitochondrial dysfunction motor neuron degeneration mouse model mutant novel prevent public health relevance therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Amyotrophic Lateral Sclerosis (ALS) is a devastating neurodegenerative disease of the motor neurons, which leads to paralysis and death within 3-5 years from diagnosis. ALS is mainly sporadic (SALS) without a known cause. Only a small fraction of ALS is familial (FALS). Thus, one of the biggest challenges in the study of the disease is how to reconcile disease mechanisms among the small percentage of familial cases and the vast majority of sporadic cases with no known etiology. It is crucial that we identify common pathogenic mechanisms between the two forms of the disease. Mitochondrial pathology is one of these common pathways, as mitochondria defects have been found in both SALS patients and transgenic mutant SOD1 (mutSOD1) mice model of ALS. Whether similar triggers in FALS and SALS damage the mitochondria is not known. Using mutSOD1 expressing cells and transgenic mice (to mimic FALS), as well as EVB immortalized lymphoblasts from SALS patients, we identified a potentially common trigger mechanism. In mutSOD1 mice, we showed that mutSOD1 aberrantly binds and forms a toxic complex with Bcl-2 in mitochondria. Upon this aberrant binding, mutSOD1 induces a conformational change in Bcl-2 that transforms it into a harmful protein by exposing the normally hidden toxic BH3 domain. Together, mutSOD1 and conformationally modified Bcl-2 impair mitochondrial viability, eventually inducing cell death. Interestingly, in a subset (~ 30%) of SALS patients with upper motor neuron onset, an oxidized form of wild type SOD1 aberrantly binds to Bcl-2, transforming Bcl- 2 into a toxic molecule through exposure of the BH3 domain, similarly to what we have reported for mutSOD1. With this competing renewal, we intend to focus on this common pathway of mitochondrial dysfunction shared by FALS-SOD1 and a subset of SALS patients. We will test in vivo the hypothesis that the conformational change in Bcl-2 leading to exposure of the toxic BH3 domain is an important mechanism in SOD1-induced mitochondrial dysfunction (AIM 1). We will then characterize the functional implications of the toxic complex between SOD1 and Bcl-2 by identifying key downstream mitochondrial target(s) (AIM 2) and determining the cellular specificity of the SOD1/Bcl-2-mediated mitochondrial dysfunction (AIM 3). Finally, we will test the beneficial effect of SOD1-like peptides that inhibit binding to Bcl-2 against SOD1-mediated cell death (AIM 4). The ultimate goal is to identify target-based therapies whose efficacy goes beyond the limited portion of familial cases.

描述（由申请人提供）：肌萎缩性侧索硬化症（ALS）是运动神经元的毁灭性神经退行性疾病，在诊断后的3 - 5年内导致瘫痪和死亡。 ALS主要是零星（萨尔），没有已知原因。只有一小部分ALS是家族性的（fals）。因此，该疾病研究的最大挑战之一是如何调和少数家族病例和绝大多数零星病因的疾病机制，没有已知的病因。至关重要的是，我们确定两种疾病形式之间的常见致病机制。线粒体病理是这些常见途径之一，因为在SALS患者和ALS的转基因突变体SOD1（MUTSOD1）小鼠模型中都发现了线粒体缺陷。尚不清楚线粒体的类似触发和萨尔的类似触发因素是否损害线粒体。我们使用表达细胞和转基因小鼠（模仿伪造）以及来自萨尔斯患者的EVB永生淋巴细胞的EVB永生的淋巴细胞，我们确定了潜在的常见触发机制。在mutsod1小鼠中，我们表明mutsod1异常结合并与线粒体中的Bcl-2形成有毒复合物。在这种异常结合后，mutsod1诱导了Bcl-2的构象变化，该变化通过暴露正常隐藏的有毒BH3结构域将其转化为有害蛋白质。 MUTSOD1和构象修饰的Bcl-2一起损害了线粒体的生存能力，最终诱导细胞死亡。有趣的是，在一个上部运动神经元发作的萨尔盐患者中（约30％）中，一种氧化的野生型SOD1形式异常结合了Bcl-2，将BCl-2转化为有毒分子，通过暴露BH3域，与我们报道的MUTSOD1相似。通过这种竞争的续约，我们打算专注于FALS-SOD1和一部分SALS患者共享的线粒体功能障碍的常见途径。我们将在体内检验一个假设，即Bcl-2的构象变化导致有毒BH3结构域暴露是SOD1诱导的线粒体功能障碍的重要机制（AIM 1）。然后，我们将通过识别下游线粒体靶标（AIM 2）并确定SOD1/BCl-2介导的线粒体功能障碍的细胞特异性来表征SOD1和BCL-2之间有毒复合物的功能含义。最后，我们将测试SOD1样肽的有益作用，该肽抑制与SOD1介导的细胞死亡的结合（AIM 4）。最终目标是确定基于目标疗法的疗效超出了家族病例的有限部分。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

ALS-linked mutant SOD1 damages mitochondria by promoting conformational changes in Bcl-2.

DOI：
10.1093/hmg/ddq202
发表时间：
2010-08-01
期刊：
Human molecular genetics
影响因子：
3.5
作者：
Pedrini S;Sau D;Guareschi S;Bogush M;Brown RH Jr;Naniche N;Kia A;Trotti D;Pasinelli P
通讯作者：
Pasinelli P

In vivo and in vitro determination of cell death markers in neurons.

神经元细胞死亡标志物的体内和体外测定。

DOI：
10.1007/978-1-61779-328-8_2
发表时间：
2011
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
0
作者：
Naniche,Nicole;Sau,Daniela;Pasinelli,Piera
通讯作者：
Pasinelli,Piera

ABC transporter-driven pharmacoresistance in Amyotrophic Lateral Sclerosis.

DOI：
10.1016/j.brainres.2014.08.060
发表时间：
2015-05-14
期刊：
BRAIN RESEARCH
影响因子：
2.9
作者：
Jablonski, Michael;Miller, David S.;Pasinelli, Piera;Trotti, Davide
通讯作者：
Trotti, Davide

Small peptides against the mutant SOD1/Bcl-2 toxic mitochondrial complex restore mitochondrial function and cell viability in mutant SOD1-mediated ALS.

针对突变型 SOD1/Bcl-2 毒性线粒体复合物的小肽可恢复突变型 SOD1 介导的 ALS 中的线粒体功能和细胞活力。

DOI：
10.1523/jneurosci.5385-12.2013
发表时间：
2013
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
0
作者：
Tan,Wenzhi;Naniche,Nicole;Bogush,Alexey;Pedrini,Steve;Trotti,Davide;Pasinelli,Piera
通讯作者：
Pasinelli,Piera

Voltage-dependent inwardly rectifying potassium conductance in the outer membrane of neuronal mitochondria.

神经元线粒体外膜中电压依赖性内向整流钾电导。

DOI：
10.1074/jbc.m110.131243
发表时间：
2010
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Fieni,Francesca;Parkar,Anjum;Misgeld,Thomas;Kerschensteiner,Martin;Lichtman,JeffW;Pasinelli,Piera;Trotti,Davide
通讯作者：
Trotti,Davide