Structure-Function Analysis of Breast Cancer-Associated Mutations in ADAM12

ADAM12 中乳腺癌相关突变的结构功能分析

• 批准号: 7940388
• 负责人: Anna Zolkiewska
• 金额: $ 44.4万
• 依托单位: KANSAS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-02 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7940388
• 关键词: Acute; Biochemical; Biological Assay; Breast Cancer Cell; Cancer Patient; Catalytic Domain; Cell Surface Proteins; Cell membrane; Cell surface; Cells; Chemicals; Co-Immunoprecipitations; Disintegrin Domain; Disintegrins; Dominant-Negative Mutation; Drosophila genus; Endoplasmic Reticulum; Extracellular Domain; Family; Genes; Human; In Vitro; Label; Lead; Link; Malignant Neoplasms; Mammary Neoplasms; Membrane; Membrane Proteins; Metalloproteases; Molecular Chaperones; Molecular and Cellular Biology; Mutate; Mutation; Patients; Pattern; Performance; Proteins; Proteolytic Processing; Proteome; Proteomics; Quality Control; Recombinants; Relative (related person); Research; Somatic Mutation; Stimulus; Structure; System; Systems Biology; TP53 gene; Testing; Training; Western Blotting; cancer cell; cancer therapy; chaperone machinery; comparative; endoplasmic reticulum stress; graduate student; human disease; malignant breast neoplasm; member; mutant; protein degradation; protein folding; public health relevance; research study; response; tool; trafficking

DESCRIPTION (provided by applicant): Metalloprotease disintegrin ADAM12 is emerging as an important breast cancer-related gene and a potential target for breast cancer therapies. Two somatic mutations, D301H in the metalloprotease domain and G479E in the disintegrin domain, are present in ~10% of primary breast tumors. We found that the D/H and G/E mutations lead to the retention of the nascent, inactive ADAM12 in the endoplasmic reticulum (ER) and a loss of active ADAM12 at the cell surface. The D/H and G/E mutants have a dominant-negative effect on wild-type ADAM12. The mechanisms responsible for the altered trafficking of the D/H and G/E mutants are not known. The main objectives of this proposal are to understand why the breast cancer-associated ADAM12 mutants are retained in the ER and what the consequences are of ADAM12 mislocalization in cancer cells. Our main hypothesis is that the D/H and G/E mutants are misfolded and retained by the ER quality control system, leading to ER stress and/or changing the repertoire of proteins secreted/shed to medium and the pattern of membrane-anchored proteins. The Specific Aims of this proposal are: 1. Determine why the D/H and G/E ADAM12 mutants are retained in the endoplasmic reticulum, and test approaches to restore the wild-type ADAM12 functionality to these mutants. 2. Assess the effect of the D/H and G/E ADAM12 mutants on the functional performance of the ER. 3. Compare the secretome and cell surface proteome of breast cancer cells expressing the wild-type ADAM12 and the D/H and G/E ADAM12 mutants. These studies will uncover the mechanism by which the breast cancer associated mutations change the function of ADAM12 in cancer cells. PUBLIC HEALTH RELEVANCE: This project is focused on the understanding the cause of malfunction of ADAM12 mutants that are frequently found in human breast tumors. The results of our studies will have a strong impact on breast cancer therapies tailored to patients harboring mutations in the ADAM12 gene.

描述（由申请人提供）：金属蛋白酶解整合素 ADAM12 正在成为一种重要的乳腺癌相关基因和乳腺癌治疗的潜在靶点。约 10% 的原发性乳腺肿瘤中存在两种体细胞突变，即金属蛋白酶结构域中的 D301H 和解整合素结构域中的 G479E。我们发现 D/H 和 G/E 突变导致新生的、无活性的 ADAM12 在内质网 (ER) 中保留，而活性 ADAM12 在细胞表面丢失。 D/H 和 G/E 突变体对野生型 ADAM12 具有显性负效应。导致 D/H 和 G/E 突变体运输改变的机制尚不清楚。该提案的主要目的是了解为什么乳腺癌相关的 ADAM12 突变体保留在 ER 中，以及 ADAM12 在癌细胞中错误定位会产生什么后果。我们的主要假设是 D/H 和 G/E 突变体被错误折叠并被 ER 质量控制系统保留，导致 ER 应激和/或改变分泌/脱落到培养基的蛋白质库和膜锚定蛋白的模式。该提案的具体目标是： 1. 确定为什么 D/H 和 G/E ADAM12 突变体保留在内质网中，并测试恢复这些突变体野生型 ADAM12 功能的方法。 2. 评估 D/H 和 G/E ADAM12 突变体对 ER 功能性能的影响。 3. 比较表达野生型 ADAM12 和 D/H 和 G/E ADAM12 突变体的乳腺癌细胞的分泌组和细胞表面蛋白质组。这些研究将揭示乳腺癌相关突变改变癌细胞中 ADAM12 功能的机制。 公共健康相关性：该项目的重点是了解人类乳腺肿瘤中常见的 ADAM12 突变体功能障碍的原因。我们的研究结果将对针对 ADAM12 基因突变患者的乳腺癌治疗产生重大影响。

项目成果

Force-induced unfolding simulations of the human Notch1 negative regulatory region: possible roles of the heterodimerization domain in mechanosensing.

人类 Notch1 负调控区的力诱导展开模拟：异二聚化结构域在机械传感中的可能作用。

• 发表时间: 2011
• 影响因子: 3.7
• 作者: Chen, Jianhan;Zolkiewska, Anna
• 通讯作者: Zolkiewska, Anna

Hesr1 and Hesr3 are essential to generate undifferentiated quiescent satellite cells and to maintain satellite cell numbers.

Hesr1 和 Hesr3 对于产生未分化的静止卫星细胞和维持卫星细胞数量至关重要。

• 发表时间: 2011-11
• 作者: Fukada, So;Yamaguchi, Masahiko;Kokubo, Hiroki;Ogawa, Ryo;Uezumi, Akiyoshi;Yoneda, Tomohiro;Matev, Miroslav M;Motohashi, Norio;Ito, Takahito;Zolkiewska, Anna;Johnson, Randy L;Saga, Yumiko;Miyagoe;Tsujikawa, Kazutake;Takeda
• 通讯作者: Takeda