Quantitative proteomic identification of novel, bioactive substrates of matrix metalloproteinase (MMP)-11 and system-wide identification of new proteases involved in breast cancer progression and metastasis

基质金属蛋白酶 (MMP)-11 新型生物活性底物的定量蛋白质组学鉴定以及参与乳腺癌进展和转移的新蛋白酶的全系统鉴定

• 批准号: 34099195
• 负责人: Dr. Ulrich auf dem Keller
• 金额: --
• 依托单位: Institut für Zellbiologie
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2006
• 资助国家: 德国
• 起止时间: 2005-12-31 至 2008-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/34099195?language=en
• 关键词: Quantitative; proteomic; identification; novel; bioactive

Rather than just cleaving extracellular matrix as their name suggests, proteolytic enzymes such as matrix metalloproteinases (MMPs) modulate angiogenesis, growth, invasion, metastasis, and phenotypic evolution of cancer cells by the irreversible processing of bioactive proteins and signaling molecules. One member of the MMP family, MMP-11, was first discovered in the reactive stroma of invasive breast carcinoma, and since then in over 200 publications is very strongly correlated with poor patient outcome in breast carcinoma. But despite extensive characterization, MMP-11 has no known natural substrates to account for its pivotal activities in breast cancer. Therefore, I will employ novel, systemwide quantitative proteomic technologies developed in the laboratory of Prof. Christopher Overall to identify and characterize bioactive substrates of MMP-11 in breast carcinogenesis. To understand the role of proteases in breast cancer development and metastasis it is not only important to define the substrate spectrum of an individual protease but also to identify and characterize new proteases involved in these processes. Therefore, in a second project I will determine the protease degradome, that is the expression levels of all proteases in a murine breast cancer model. For this purpose, an oligonucleotide microarray termed the CLIP-CHIPTM comprising all known human and murine proteases, inactive homologues and inhibitors that was designed and made in the host laboratory will be used. Interesting proteases that are highly expressed in cancer or reactive stromal tissues will be analyzed for their spectrum of substrates termed the substrate degradome. These experiments will provide further insight into the role of proteases including matrix metalloproteinases in cancer progression, especially how they modify the activity of drug target substrates and so contribute to drug target validation in breast cancer.

蛋白水解酶如基质金属蛋白酶（MMP）通过生物活性蛋白和信号分子的不可逆加工来调节癌细胞的血管生成、生长、侵袭、转移和表型进化，而不是如其名称所暗示的那样仅切割细胞外基质。MMP家族的一个成员MMP-11首先在浸润性乳腺癌的反应性基质中被发现，并且从那时起在超过200篇出版物中与乳腺癌中的不良患者结局非常密切相关。但是，尽管有广泛的表征，MMP-11没有已知的天然底物来解释其在乳腺癌中的关键活性。因此，我将采用Christopher Overall教授实验室开发的新的全系统定量蛋白质组学技术来鉴定和表征MMP-11在乳腺癌发生中的生物活性底物。为了了解蛋白酶在乳腺癌发展和转移中的作用，不仅重要的是要确定单个蛋白酶的底物谱，而且要鉴定和表征参与这些过程的新蛋白酶。因此，在第二个项目中，我将确定蛋白酶降解组，即小鼠乳腺癌模型中所有蛋白酶的表达水平。为此目的，将使用在宿主实验室中设计和制备的称为CLIP-CHIPTM的寡核苷酸微阵列，其包含所有已知的人和鼠蛋白酶、无活性同源物和抑制剂。将分析在癌症或反应性基质组织中高度表达的感兴趣的蛋白酶的底物谱，称为底物降解组。这些实验将进一步深入了解蛋白酶（包括基质金属蛋白酶）在癌症进展中的作用，特别是它们如何修饰药物靶底物的活性，从而有助于乳腺癌中的药物靶点验证。