Stabilizing fALS SOD1 Variants by Crosslinking Subunits

描述(申请人提供)：编码SOD1基因的120多个突变与肌萎缩侧索硬化症有关。SOD1突变会导致毒性，一个流行的假设是，功能的获得涉及SOD1二聚体的解离和聚集。因此，稳定SOD1二聚体成为SOD1二聚体治疗的一种途径。我们已经发现了一种通过将Cys111残基与共价交联剂捆绑在一起来稳定SOD1二聚体的特定的化学计量方法。Cys111是位于二聚体界面的表面残基，距离相对单体上的Cys111(8？)只有一小段距离。我们的假设是，桥接Cys111残基的化合物将作为新的药理伴侣发挥作用，稳定Fals SOD1变体，并且基于多肽的交联剂适合于开发可用于创建第二代改进多肽的定量结构活性模型。为了激发人们的兴趣，我们的第一个目标是扩大我们最初发现的范围，将最流行的五种FAL变体包括在内。虽然我们的第一代交联剂在体外效果良好，但它们在体内是有毒的。因此，我们的第二个目标是创造一种新的基于多肽的化学物质，这种化学物质经过优化可以使Cys111交联。 公共卫生相关性：包括肌萎缩侧索硬化症在内的神经退行性疾病已被证明非常难以治疗，例如，肌萎缩侧索硬化症只有一种FDA批准的药物用于治疗，报告对患者生存和生活质量几乎没有影响。直到最近，研究人员还不了解肌萎缩侧索硬化症相关蛋白的毒性机制，但现在许多人认为，这涉及到蛋白质SOD1从一个二聚体解离成两个单体。防止这种解离已成为治疗的重点，我们已经发现了一种新颖而有效的体外稳定SOD1的策略，该策略可以使单个单体交联。这项提议旨在通过测试最流行的SOD1变体来扩大我们最初发现的范围，并开发和优化比我们第一代交联剂毒性更小的分子。

DESCRIPTION (provided by applicant): Over 120 mutations in the gene encoding SOD1 are associated with amyotrophic lateral sclerosis. SOD1 mutations result in a toxic property, and a prevailing hypothesis is that a gain-of-function involves dissociation of the SOD1 dimer followed by aggregation. As a result, stabilization of the SOD1 dimer has become an approach for SOD1 therapy. We have discovered a specific and stoichiometric method for stabilizing the SOD1 dimer by tethering Cys111 residues with a covalent cross-linker. Cys111 is a surface residue situated at the dimer interface, and only a short distance apart from the Cys111 (8¿) on an opposing monomer. Our hypothesis is that compounds that bridge Cys111 residues will function as novel pharmacological chaperones that stabilize fALS SOD1 variants and that peptide-based cross-linkers are suitable for developing a quantitative structure activity model that can be used to create a second generation of improved peptides. In the hopes of inspiring interest, our first aim is to expand the scope of our original findings to include the five most prevalent fALS variants. While our first generation cross-linkers work well in vitro, they known to be toxic in vivo. Therefore, our second aim is to create a new class of peptide-based chemicals that are optimized to cross-link Cys111. PUBLIC HEALTH RELEVANCE: The neurodegenerative diseases, including ALS, have proven extraordinarily difficult to treat, for example, ALS has only one FDA-approved drug for its treatment, reporting little or no effect on patient survival and quality of life. Until recently researchers did not understand the mechanism of ALS- associated proteins' toxicity, but now many believe this involves the dissociation of the protein SOD1 from a dimer into two monomers. Preventing this dissociation has become a focus for therapy, and we have discovered a novel and effective in vitro strategy for stabilizing SOD1 that cross-links individual monomers. This proposal aims to expand the scope of our original findings by testing the most prevalent SOD1 variants, and to develop and optimize molecules that are less toxic than our first generation of cross-linkers.