Genetic and Functional Analysis of Drosophila Mmp 1

描述（由申请人提供）： 基质金属蛋白酶（MMP）是一类在体外降解细胞外基质的酶，并且在癌症、炎症和其他人类病理中上调。虽然抑制剂研究已经在临床试验中进行了测试，但它们对癌症无效，并且具有不可接受的副作用。这些失败的试验表明，在设计有效的治疗方法之前，科学界需要更全面地了解MMP生物学及其正常的生理作用：它们的功能是什么以及它们切割什么底物。然而，由于小鼠中有22种MMPs，并且它们可以补偿彼此的损失，因此小鼠中的突变分析不能提供有关功能的完整信息。果蝇（Drosophila melanogaster）是研究MMPs的理想生物，因为它们的基因组中只有两种MMPs。先前在两个基因中产生突变，并且突变体具有特定的表型，表明基因是非冗余的。在初步研究中，双杂交筛选确定了特异性结合dMmp 1血红素结合蛋白结构域的候选蛋白，并且与一个候选蛋白的物理相互作用已通过免疫共沉淀得到证实。在特定目标1中，将在培养、体外和体内确定该候选物与dMmp 1之间的关系;在初步研究中，它似乎以Mmp 1特异性方式被蛋白水解。Mmp 1和候选相互作用物可能对动物对脓毒性创伤的反应很重要，它们在脓毒性伤口愈合中的作用将在具体目标2中确定。为了更好地了解MMP在体内的功能，将在特定目的3中鉴定气管系统中dMmp 1及其C末端血红素结合蛋白结构域的细胞功能。 (for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。其中一个蛋白质家族，称为MMPs，已被制药公司用于设计抗癌药物，但这些药物具有严重的副作用，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式生物中确定MMP的功能和机制，希望能够学习如何制造更好的癌症药物。

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.