Unraveling the Role of MMP3 in the Cisplatin Resistance of Ovarian Cancer

摘要 这项资助申请的目的是为了促进代表不足的人在生物医学研究方面的卓越表现。 波多黎各大学医学院校区的研究生和本科生。世界范围内， 与癌症相关的死亡人数继续增加，原因是他们有能力对化疗产生抗药性和 转移瘤。对于患有卵巢癌的女性来说，70%的人会对一线治疗产生抵抗力， 顺铂。虽然顺铂的耐药机制已经被广泛研究，但还没有有效的治疗方法。 结果，使这样的研究变得至关重要和重要。在寻找新的治疗策略以逆转 在这些可怕的趋势中，我们发现基质金属蛋白酶3(MMP3)在顺铂中高度丰富 使用差异基因表达研究，将耐药卵巢癌细胞与有反应的卵巢癌细胞进行比较。我们的 种子研究结果进一步表明，MMP3水平高的卵巢癌肿瘤复发速度更快 这进一步支持了这种酶在疾病中发挥关键作用的观点 进步。MMP3属于MMPs蛋白水解酶家族，可降解MMPs中的多种成分 细胞外基质(ECM)。MMPs的蛋白质结构包括催化区和血凝素(HPX)。 域。已经设计了几种合成的和天然的基质金属蛋白酶抑制剂来抑制其催化结构域 MMPs。然而，这些研究几年前就被放弃了，因为非 这些抑制物的特定特征。然而，来自我们实验室的令人兴奋的数据试图逆转这些 令人失望的趋势表明，MMP3靶向小干扰RNA(SiRNAs)显著 降低顺铂耐药卵巢癌细胞的细胞增殖和侵袭能力。我们没有 当我们使用一种结合到催化结构域的抑制物时，观察到增殖或侵袭性的减少 MMP3。我们的初步结果与MMP3的HPX结构域结合的证据是一致的 连接细胞外基质和细胞内分子通路的蛋白质。因此，我们准备研究不 MMP3不仅在顺铂耐药中起作用，而且还能识别可能的相关蛋白 它们本身就是研究的新的治疗目标。因此， 我们计划验证我们的假设，即MMP3(HPX结构域)的其他区域与促进顺铂的蛋白质相互作用 抵抗。结合使用分子生物学工具和我们广泛而成熟的卵巢 癌症模型，我们将用以下具体目标检验这一中心假说：(1)确定MMP3是否 在卵巢癌模型中调节对化疗药物顺铂的敏感性，(2)鉴定蛋白质 在顺铂耐药的卵巢癌细胞中直接与MMP3相互作用，以及(3)确定其生物学特性 SiRNA介导的MMP3靶向在卵巢癌模型中的作用和治疗效果。结果： 这项研究项目有可能通过加强对治疗耐药性的理解而产生影响。 MMP3在卵巢癌中的非催化作用。

Abstract The purpose of this grant application is to promote the excellence in biomedical research of underrepresented graduate and undergraduate students of the University of Puerto Rico Medical Sciences Campus. World-wide, cancer related deaths continue to increase due to their ability to become chemotherapy resistant and metastasize. For women with ovarian cancer, a staggering 70% will become resistant to the front-line therapy, cisplatin. While the mechanism of cisplatin resistance has been extensively studied, no effective treatments have resulted, making such research critical and important. During the search for new therapeutic strategies to reverse these horrendous trends, we identified matrix metalloproteinase 3 (MMP3) to be highly abundant in cisplatin resistant ovarian cancer cells, as compared to responsive ones, using differential gene expression studies. Our seminal findings have further demonstrated that ovarian cancer tumors with high MMP3 levels relapse at a faster rate than those expressing lower levels, further supporting the notion that this enzyme plays a key role in disease progression. MMP3 belongs to the MMPs family of proteolytic enzymes that degrade multiple components of the extracellular matrix (ECM). The protein structure of MMPs includes a catalytic domain and a hemopexin (HPX) domain. Several synthetic and natural MMP inhibitors have been designed to inhibit the catalytic domain of MMPs. However, these studies were abandoned years ago due to the overt toxicities resulting from the non- specific profile of those inhibitors. However, exciting data from our laboratory seeks to reverse these disappointing trends by demonstrating that MMP3-targeting small-interfering RNAs (siRNAs) significantly reduced cell proliferation and the invasiveness ability of cisplatin resistant ovarian cancer cells. We did not observe reduced proliferation or invasiveness when we used an inhibitor that binds to the catalytic domain of the MMP3. Our preliminary results are in agreement with evidence that the HPX domain of MMP3 could binds to proteins connecting the ECM with intracellular molecular pathways. Therefore, we are poised to examine not only the contribution of MMP3 to the cisplatin resistance, but also to identify associated proteins that might contribute to this disease and they themselves represent new therapeutic targets for investigation. Therefore, we plan to test our hypothesis that other regions of MMP3 (HPX domain) interact with proteins promoting cisplatin resistance. Using a combination of molecular biology tools and our extensive and well established ovarian cancer models, we will test this central hypothesis with the following specific aims: (1) determine whether MMP3 in ovarian cancer models mediates susceptibility to the chemotherapy agent cisplatin, (2) identify the protein directly interacting with MMP3 in cisplatin resistant ovarian cancer cells, and (3) determine the biological consequences and therapeutic effects of siRNA-mediated MMP3 targeting in ovarian cancer models. Results of this research project have the potential to provide impact by enhancing understanding of therapy resistance and non-catalytic functions of MMP3 in ovarian cancer.