Genetic and Functional Analysis of Drosophila Mmp 1

果蝇 Mmp 1 的遗传和功能分析

• 批准号: 7036004
• 负责人: Andrea W Page-McCaw
• 金额: $ 27.49万
• 依托单位: RENSSELAER POLYTECHNIC INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-25 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7036004
• 关键词: Drosophilidae; antibody; arthropod genetics; cell adhesion; cell migration; collagenase; developmental genetics; enzyme mechanism; enzyme substrate; gene interaction; immune response; immunologic assay /test; intracellular; larva; membrane proteins; protein localization; protein protein interaction; septicemia; trachea; transposon /insertion element; wound healing

DESCRIPTION (provided by applicant): Matrix metalloproteinases (MMP) are a class of enzymes that degrade extracellular matrix in vitro and are upregulated in cancer, inflammation, and other human pathologies. Although inhibitor studies have been tested in clinical trials, they have not been effective against cancer and have had unacceptable side effects. These failed trials indicate that before effective therapies can be designed, the scientific community needs a more complete understanding of MMP biology and their normal physiological roles: what their functions are and what substrates they cleave. However, because there are 22 MMPs in mice and they can compensate for each other's loss, mutant analysis in mice does not give complete information about function. The fruitfly, Drosophila melanogaster, is an ideal organism for the study of MMPs, because they have only two MMPs in their genome. Previously mutations were generated in both genes, and the mutants have specific phenotypes indicating that the genes are non-redundant. In preliminary studies, two-hybrid screening identified candidate proteins that specifically bind to the dMmp1 hemopexin domain, and the physical interaction with one candidate has been confirmed by co-immunoprecipitation. In Specific Aim 1, the relationship between this candidate and dMmp1 will be determined in culture, in vitro, and in vivo; in preliminary studies, it appears to be proteolyzed in an Mmp1 -specific manner. Both Mmp1 and the candidate interactor may be important for the animal's response to septic wounding, and their roles in septic wound healing will be determined in Specific Aim 2. To gain a better understanding of MMP function in vivo, the cellular functions of the dMmp1 and its C-terminal hemopexin domain will be identified in the tracheal system in Specific Aim 3. (for lay people:) In order to grow and metastasize, cancers are believed to rely on proteins that break down and remodel body tissues. One such protein family, called MMPs, has been targeted by pharmaceutical companies for designing anti-cancer drugs, but these have had crippling side effects because we do not understand what MMPs normally do in the body. We will identify the functions and mechanisms of an MMP in a simple model organism in the hopes of learning how to make better cancer drugs.

描述（由申请人提供）： 基质金属蛋白酶（MMP）是一类酶，在体外降解细胞外基质，并在癌症，炎症和其他人类病理中上调。尽管抑制剂研究已经在临床试验中进行了测试，但它们对癌症没有有效，并且具有不可接受的副作用。这些失败的试验表明，在设计有效的疗法之前，科学界需要对MMP生物学及其正常生理角色有更全面的了解：它们的功能是什么以及它们切割的底物。但是，由于小鼠有22个MMP，并且可以弥补彼此的损失，因此小鼠突变分析不会提供有关功能的完整信息。果蝇（果蝇）果蝇（Drosophila Melanogaster）是研究MMP的理想生物，因为它们的基因组中只有两个MMP。以前在两个基因中都产生了突变，突变体具有特定的表型，表明这些基因是非冗余的。在初步研究中，两种杂交筛查确定了特异性结合DMMP1呼吸蛋白结构域的候选蛋白，并且通过共免疫沉淀证实了与一个候选者的物理相互作用。在特定的目标1中，该候选者与DMMP1之间的关系将在培养物，体外和体内确定。在初步研究中，它似乎是以MMP1特异性方式进行的。 MMP1和候选者的相互作用对于动物对败血性伤口的反应可能很重要，并且将在特定的目标2中确定它们在败血症伤口愈合中的作用。为了更好地了解体内MMP功能，DMMP1的细胞功能，DMMP1的细胞功能及其C-末端溶血结构域的细胞功能将在特定特定目标中的气管系统中鉴定出来。 （对于外行人:)为了生长和转移，癌症被认为依赖于分解和重塑体组织的蛋白质。这样的蛋白质家族称为MMP，是制药公司针对设计抗癌药物的目标，但是这些蛋白质家族具有严重的副作用，因为我们不了解MMPS在体内通常会做什么。我们将确定MMP在简单模型生物体中的功能和机制，以期学习如何制造更好的癌症药物。