Genetic and Functional Analysis of Drosophila Mmp 1

描述(由申请人提供)： 基质金属蛋白酶(MMPs)是一类在体外降解细胞外基质的酶，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，但它们对癌症并不有效，而且产生了不可接受的副作用。这些失败的试验表明，在设计有效的治疗方法之前，科学界需要更全面地了解基质金属蛋白酶的生物学及其正常的生理作用：它们的功能是什么，它们分解的底物是什么。然而，由于小鼠体内有22个MMPs，它们可以弥补彼此的损失，因此对小鼠的突变分析并不能给出关于功能的完整信息。果蝇是研究MMPs的理想生物，因为它们的基因组中只有两种MMPs。此前，这两个基因都产生了突变，并且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，双杂交筛选确定了与dMmp1血凝蛋白结构域特异结合的候选蛋白，并通过免疫共沉淀证实了与一个候选蛋白的物理相互作用。在特定目标1中，该候选基因与dMmp1之间的关系将在培养、体外和体内确定；在初步研究中，它似乎以MMP1特异的方式被蛋白质降解。MMP1和候选的相互作用因子在动物对脓毒症创伤的反应中都可能是重要的，它们在脓毒症伤口愈合中的作用将在特定的目标2中确定。为了更好地了解MMP1在体内的功能，我们将在特定的目标3中确定dMmp1及其C末端血凝蛋白结构域在气管系统中的细胞功能。 (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身体组织的蛋白质。其中一种名为MMPs的蛋白质家族因设计抗癌药物而成为制药公司的目标，但这些药物产生了严重的副作用，因为我们不了解MMPs通常在体内起什么作用。我们将在一个简单的模型生物体中确定基质金属蛋白酶的功能和机制，以期学习如何制造更好的抗癌药物。

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.