Mechanisms of Aggregation in Light Chain Amyloidosis

轻链淀粉样变性的聚集机制

• 批准号: 8665966
• 负责人: Marina Ramirez-Alvarado
• 金额: $ 31.11万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8665966
• 关键词: Affect; Aftercare; Amino Acid Sequence; Amyloid; Amyloid Fibrils; Amyloid Proteins; Amyloidosis; Biological Assay; Blood Circulation; Bone Marrow; Cardiac; Cardiac Myocytes; Caspase; Cell Survival; Cessation of life; Clinical; Collaborations; Cross Circulation; Data; Deposition; Development; Disease; Disease Progression; Drug Design; Equipment; Funding; Future; Genetic Polymorphism; Goals; Human; Immunoglobulins; Kinetics; Knowledge; Laboratories; Light; Light-Chain Immunoglobulins; Malignant - descriptor; Nature; Organ; Organ failure; Patients; Peptide Sequence Determination; Plasma Cells; Play; Population; Process; Property; Proteins; Qualifying; Reagent; Recruitment Activity; Reporting; Role; Seeds; Somatic Mutation; Structure; Survival Rate; Testing; Therapeutic; Thermodynamics; Toxic effect; Work; amyloid formation; arteriole; base; cytotoxic; cytotoxicity; cytotoxicity test; design; dimer; innovation; novel; primary amyloidosis of light chain type; protein misfolding; protein structure; therapeutic development

DESCRIPTION (provided by applicant): The overall goal of this proposal is to identify the misfolded cytotoxic species populated by light chain amyloidosis (AL) proteins. AL is a rare, fatal misfolding disease characterized by the proliferation of monoclonal plasma cells that secrete immunoglobulin light chains that misfold as amyloid deposits in vital organs. Current treatments reduce the population of the malignant monoclonal plasma cells. Little is known about the misfolding process converting soluble proteins into insoluble AL fibrils and the nature of the toxic species generated during this process; identifying the toxic species is crucial to developing targeted therapies for AL. We have determined that the protein AL-09 forms an altered dimer interface. A single somatic mutation is responsible for the change in the dimer interface, causing loss of thermodynamic stability, and promoting amyloid formation with respect to its germline ('wild type') protein. Our preliminary data show that AL-09 is the most amyloidogenic AL protein studied in our laboratory, it is highly toxic to cardiomyocytes in its soluble form, and its amyloid fibril core is consistent with the altered dimer interface structure. In addition, we want to explore alternative mechanisms of cytotoxicity that would explain the phenomenon observed with some patients who have achieved significant reduction of circulating light chain post-treatment but still have disease progression. Based on this information, our central hypothesis is that soluble AL light chain species are highly toxic. There are different mechanisms that generate AL toxic species. Specific somatic mutations in AL light chains cause the protein to favor one or more cytotoxic mechanisms. In aim 1, we will test the hypothesis that protein internalization is required for cytotoxicity using toxicity assays on cardiomyocytes and human arteriole dilation assays. For aim 2, we will test the hypothesis that low thermodynamic stability of some AL fibrils could cause disaggregation that may play a role in cytotoxicity. In addition, we will identify alternative mechanisms of cytotoxicity by testing the hypothesis that some AL proteins are able to cross seed with non-pathogenic light chains present in the normal repertoire of immunoglobulin molecules. For aim 3, we will test the hypothesis that AL fibril core composition is mostly driven by the predominant dimer structure populated by each AL protein and will correlate to protein cytotoxicity. Our results will have a collective impact because they will provide new knowledge into the mechanism of AL by identifying common cytotoxic species and AL fibril core regions. This knowledge will be essential for the development of rational drug design to ultimately eliminate the devastating consequences of this disease.

描述(由申请人提供)：这项提案的总体目标是鉴定由轻链淀粉样变性(AL)蛋白填充的错误折叠的细胞毒性物种。阿尔茨海默病是一种罕见的致命的错误折叠疾病，其特征是分泌免疫球蛋白轻链的单克隆浆细胞增殖，这些轻链错误折叠为重要器官中的淀粉样沉积。目前的治疗方法减少了恶性单克隆性浆细胞的数量。关于将可溶性蛋白质转化为不溶性AL纤维的错误折叠过程以及在此过程中产生的有毒物种的性质，人们知之甚少；识别有毒物种对于开发针对AL的靶向治疗至关重要。我们已经确定，蛋白质AL-09形成了一个改变的二聚体界面。一个单一的体细胞突变导致二聚体界面的变化，导致热力学稳定性的丧失，并促进相对于其种系(‘野生型’)蛋白的淀粉样蛋白的形成。我们的初步数据显示，AL-09是我们实验室研究的最具淀粉样变性的AL蛋白，它以可溶性形式对心肌细胞具有高度毒性，其淀粉样原纤维核心与改变的二聚体界面结构一致。此外，我们希望探索细胞毒性的替代机制，以解释一些患者在治疗后循环轻链显著减少但仍有疾病进展的现象。基于这一信息，我们的中心假设是，可溶性AL轻链物种具有高度毒性。有不同的机制产生AL有毒物种。AL轻链上的特定体细胞突变导致蛋白质倾向于一种或多种细胞毒机制。在目标1中，我们将通过对心肌细胞的毒性试验和人小动脉扩张试验来验证蛋白质内化是细胞毒性所必需的假设。对于目标2，我们将检验这样的假设，即某些AL纤维的低热力学稳定性可能会导致解聚，这可能在细胞毒性中发挥作用。此外，我们将通过测试一些AL蛋白能够与正常免疫球蛋白分子库中存在的非致病轻链杂交的假设来确定细胞毒性的替代机制。对于目标3，我们将检验这样的假设，即AL原纤维核心成分主要由每个AL蛋白填充的主要二聚体结构驱动，并将与蛋白质的细胞毒性相关。我们的结果将产生集体影响，因为它们将通过识别常见的细胞毒性物种和AL原纤维核心区为AL的发病机制提供新的知识。这些知识对于开发合理的药物设计以最终消除这种疾病的破坏性后果至关重要。

项目成果

Monosialoganglioside-Containing Nanoliposomes Restore Endothelial Function Impaired by AL Amyloidosis Light Chain Proteins.

• DOI: 10.1161/jaha.116.003318
• 发表时间: 2016-06-13
• 期刊: Journal of the American Heart Association
• 影响因子: 5.4
• 作者: Franco DA;Truran S;Weissig V;Guzman-Villanueva D;Karamanova N;Senapati S;Burciu C;Ramirez-Alvarado M;Blancas-Mejia LM;Lindsay S;Hari P;Migrino RQ
• 通讯作者: Migrino RQ

Principles of protein misfolding.

蛋白质错误折叠的原理。

• DOI: 10.1016/s0079-6603(08)00404-2
• 发表时间: 2008
• 期刊: Progress in molecular biology and translational science
• 作者: Ramirez-Alvarado,Marina

Kinetic control in protein folding for light chain amyloidosis and the differential effects of somatic mutations.

• DOI: 10.1016/j.jmb.2013.10.016
• 发表时间: 2014-01-23
• 期刊: Journal of molecular biology
• 影响因子: 5.6
• 作者: Blancas-Mejía LM;Tischer A;Thompson JR;Tai J;Wang L;Auton M;Ramirez-Alvarado M
• 通讯作者: Ramirez-Alvarado M

Nanoliposomes protect against AL amyloid light chain protein-induced endothelial injury.

• DOI: 10.3109/08982104.2013.838258
• 发表时间: 2014-03
• 期刊: Journal of liposome research
• 影响因子: 4.4
• 作者: Truran S;Weissig V;Ramirez-Alvarado M;Franco DA;Burciu C;Georges J;Murarka S;Okoth WA;Schwab S;Hari P;Migrino RQ
• 通讯作者: Migrino RQ

Structural alterations within native amyloidogenic immunoglobulin light chains.

• DOI: 10.1016/j.jmb.2009.04.010
• 发表时间: 2009-05-29
• 期刊: JOURNAL OF MOLECULAR BIOLOGY
• 影响因子: 5.6
• 作者: Randles, Edward G.;Thompson, James R.;Martin, Douglas J.;Ramirez-Alvarado, Marina
• 通讯作者: Ramirez-Alvarado, Marina