Probing the cellular Mechanisms of Transthyretin Cytotoxicity

甲状腺素运载蛋白细胞毒性的细胞机制探讨

基本信息

批准号：
8680097
负责人：
Natalia Reixach
金额：
$ 37.42万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8680097
关键词：
Abbreviations Accounting Affect Affinity Aging Alzheimer&apos s Disease Amyloid Amyloid Fibrils Amyloidosis Animal Model Antioxidants Apoptotic Biochemical Cardiomyopathies Cell Death Cell Line Cells Cellular Assay Cerebrospinal Fluid Data Deposition Disease Dissociation Evaluation Event Extracellular Matrix Familial Amyloid Neuropathies Family Functional disorder Generations Glycosaminoglycans Goals HSF1 Hand Human Pathology Kinetics Knowledge Lead Light Mass Spectrum Analysis Mediating Modeling Molecular Mutate Organ Pathogenesis Pathway interactions Plants Prealbumin Predisposition Process Proteins Proteomics Reactive Oxygen Species Receptor Cell Resveratrol Retinol Binding Proteins Risk Factors Role Secondary Protein Structure Specificity System Testing Therapeutic Agents Therapeutic Intervention Thyroxine Time Tissues Transgenic Model Variant Work age related analog analytical ultracentrifugation base cell injury conformer cytotoxic cytotoxicity extracellular heat-shock factor 1 in vivo inhibitor/antagonist insight molecular marker mutant novel therapeutics oxidation polyphenol prevent protein misfolding receptor research study response small molecule tissue culture tool

项目摘要

DESCRIPTION (provided by applicant): The transthyretin (TTR) amyloidoses are fatal disorders of secondary protein structure characterized by extracellular deposition of aggregates, including amyloid fibrils, derived from the secreted transthyretin protein. Aging is a risk factor for both the senile systemic (normal sequence) and familial (mutated sequence) forms of these disorders. At this time, no pharmacologic treatment is available. Recent data suggest that oligomers generated early in the course of TTR misfolding may be critical in the in vivo pathogenesis. We have developed the cell and molecular biologic tools to precisely analyze the molecular mechanisms underlying cell damage. Such knowledge is critical to the long-term goal of finding new therapeutic strategies to treat these diseases. Our working hypotheses are 1) TTR cytotoxicity in tissue culture reflects the human pathology regarding the susceptibility of target tissues and the amyloidogenicity of different TTR variants. 2) There is a relationship between the age-related increase in protein oxidation and the onset of TTR amyloidoses. 3) The interaction between the appropiate TTR conformer and target cells results in the generation of reactive oxygen species which, in turn, trigger an apoptotic cascade leading to cell death. 4) There is a cell receptor mediating the cytotoxic response. 5) The sequence of events leading to cell death can be prevented with compounds structurally derived from the plant polyphenol resveratrol and other antioxidants. These hypotheses will be tested in our 3 specific aims. In aim 1 we will establish tissue culture systems that mimic the human pathology in terms of tissue specificity and TTR variant. With these models in hand we will precisely identify the responsible TTR cytotoxic species, and define the role of protein oxidation and of the extracellular matrix on TTR-induced cytotoxicity. In aim 2 we will characterize and identify the cell receptor involved in the cytotoxic response using biochemical, immunochemical and proteomic mass spectrometry. In aim 3 we will use the cell system to screen compounds that inhibit TTR-induced cytotoxicity to find substances that may prevent TTR amyloidosis in vivo. We will define the mechanism of inhibition of TTR- induced cytotoxicity by the active small molecules which might in turn, be applicable to other amyloidoses. The insights gained from these studies will provide lead compounds and potential new pathways for therapeutic intervention.

描述（由申请人提供）：运甲状腺素蛋白（TTR）淀粉样变性是二级蛋白质结构的致命性疾病，其特征在于聚集物的细胞外沉积，包括衍生自分泌的运甲状腺素蛋白的淀粉样原纤维。衰老是这些疾病的老年系统性（正常序列）和家族性（突变序列）形式的危险因素。目前尚无可用的药物治疗。最近的数据表明，TTR 错误折叠过程早期产生的寡聚体可能在体内发病机制中至关重要。我们开发了细胞和分子生物学工具来精确分析细胞损伤的分子机制。这些知识对于寻找治疗这些疾病的新治疗策略的长期目标至关重要。我们的工作假设是 1) 组织培养中的 TTR 细胞毒性反映了有关靶组织易感性和不同 TTR 变体的淀粉样蛋白生成性的人类病理学。 2）年龄相关的蛋白质氧化增加与TTR淀粉样变性的发生之间存在关系。 3) 适当的TTR构象异构体和靶细胞之间的相互作用导致活性氧的产生，进而引发细胞凋亡级联反应，导致细胞死亡。 4) 存在介导细胞毒性反应的细胞受体。 5) 可以使用结构上源自植物多酚白藜芦醇和其他抗氧化剂的化合物来预防导致细胞死亡的一系列事件。这些假设将在我们的 3 个具体目标中得到检验。在目标 1 中，我们将建立在组织特异性和 TTR 变异方面模仿人类病理学的组织培养系统。有了这些模型，我们将精确识别负责的 TTR 细胞毒性物质，并定义蛋白质氧化和细胞外基质对 TTR 诱导的细胞毒性的作用。在目标 2 中，我们将使用生化、免疫化学和蛋白质组质谱来表征和鉴定参与细胞毒性反应的细胞受体。在目标 3 中，我们将利用细胞系统筛选抑制 TTR 诱导的细胞毒性的化合物，以找到可能在体内预防 TTR 淀粉样变性的物质。我们将定义活性小分子抑制 TTR 诱导的细胞毒性的机制，这可能反过来适用于其他淀粉样变性。从这些研究中获得的见解将为治疗干预提供先导化合物和潜在的新途径。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Quantification of quaternary structure stability in aggregation-prone proteins under physiological conditions: the transthyretin case.

DOI：
10.1021/bi500739q
发表时间：
2014-10-21
期刊：
BIOCHEMISTRY
影响因子：
2.9
作者：
Robinson, Lei Z.;Reixach, Natalia
通讯作者：
Reixach, Natalia

Repositioning tolcapone as a potent inhibitor of transthyretin amyloidogenesis and associated cellular toxicity.

DOI：
10.1038/ncomms10787
发表时间：
2016-02-23
期刊：
Nature communications
影响因子：
16.6
作者：
Sant'Anna R;Gallego P;Robinson LZ;Pereira-Henriques A;Ferreira N;Pinheiro F;Esperante S;Pallares I;Huertas O;Almeida MR;Reixach N;Insa R;Velazquez-Campoy A;Reverter D;Reig N;Ventura S
通讯作者：
Ventura S