Furin and the Etiopathogenesis of UV-Induced Skin Cancer

弗林蛋白酶和紫外线诱发的皮肤癌的发病机制

• 批准号: 7576602
• 负责人: ANDRES J KLEIN-SZANTO
• 金额: $ 36.16万
• 依托单位: FOX CHASE CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7576602
• 关键词: Biological Process; Biomedical Engineering; Cell Proliferation; Complementary DNA; Development; Early Diagnosis; Enzymes; Epidermis; Experimental Animal Model; Family; Growth; Growth Factor; Growth Factor Receptors; Growth and Development function; Human; Impairment; Lead; MMP11 gene; Malignant Neoplasms; Matrix Metalloproteinases; Modeling; Mus; Pathogenesis; Peptide Hydrolases; Peptides; Physiological; Play; Predisposition; Premalignant; Process; Proprotein Convertase 2; Proprotein Convertases; Proteins; Protocols documentation; Public Health Applications Research; Relative (related person); Role; Series; Skin; Skin Cancer; Skin Carcinogenesis; Skin Neoplasms; Squamous cell carcinoma; Substrate Specificity; Testing; Transgenic Animals; Transgenic Mice; Transgenic Model; Transgenic Organisms; UV Radiation Exposure; UV carcinogenesis; UV induced; Ultraviolet Rays; base; cancer cell; carcinogenesis; design; in vivo; inhibitor/antagonist; keratinocyte; mouse model; neoplastic; neoplastic cell; overexpression; public health relevance; receptor; research study; synergism; tumor; tumor growth

DESCRIPTION (provided by applicant): Furin is a protein processing enzyme of the family of proprotein convertases (PCs), which has a significant role in cancer etiopathogenesis. In this application, we propose to investigate the function of furin during skin tumor development, focusing on the early and late changes that take place after UV exposure of the mouse skin. PACE4, another related PC, has been shown to have a role in murine tumor cell invasiveness by activating relevant proteases. Furin is also able to enhance cell proliferation in human precancerous and cancer cells by activating growth factor and growth factor receptors, such as IGF-R1. The lack of a mouse model to study the function of furin during carcinogenesis prompted us to design a series of experiments using transgenic mice to facilitate the better understanding of this prominent PC in the etiopathogenesis of skin cancer. Our objective in this proposal is to determine the mechanisms by which furin contributes to tumor development alone or in combination with PACE4 using transgenic mice overexpressing PCs. Two aims have been designed to this effect: First we will determine if the targeted in vivo overexpression of furin alone will enhance the development of experimental skin cancer induced by UV. We shall investigate the simultaneous in vivo overexpression of PACE4 and furin using bigenic mice in order to establish the possible cooperation of the 2 PCs in skin carcinogenesis. Substrate- specificity will be evaluated in keratinocyte cultures derived from monogenic (K5-PACE4 and K5-Furin) as well as from bigenic mice (K5-PACE4/Furin). These experiments will clarify the significance of furin's pro-proliferative effect and whether its pro-invasive effects are the same or different from PACE4's predominant pro-invasive function. In addition, this specific aim will determine whether the combination of both PCs have an enhancing effect on any or both of these biological processes. Second, we will also determine if PC inhibition blocks tumor growth and development in transgenic models. Crossing monogenic PC transgenic mice as well as double transgenics K5-Furin/PACE4 with K5-PDX and K5-PC-Propeptide transgenic mice expressing either the cDNA of a competitive PC peptide inhibitor or that of the physiological inhibitors of PCs, we will determine if inhibition of PCs alters the formation of skin tumors induced by UV carcinogenesis. PUBLIC HEALTH RELEVANCE: This application will use modern experimental animal models to dissect the function of proprotein convertases, enzymes that activate relevant cancer-associated biomolecules, during the early formation and growth of squamous cell carcinomas, one of the most common cancers of the human skin. The proposed studies will lead to the identification of mechanisms involved in the origination of cancer that could be of great practical value in its early diagnosis and treatment.

描述（由申请人提供）：弗林蛋白酶是原蛋白转化酶（PC）家族的一种蛋白质加工酶，其在癌症发病机制中具有重要作用。在本申请中，我们建议研究弗林蛋白酶在皮肤肿瘤发展过程中的功能，重点关注小鼠皮肤紫外线照射后发生的早期和晚期变化。另一种相关的 PC PACE4 已被证明通过激活相关蛋白酶在小鼠肿瘤细胞侵袭中发挥作用。弗林蛋白酶还能够通过激活生长因子和生长因子受体（例如 IGF-R1）来增强人类癌前细胞和癌细胞的细胞增殖。由于缺乏研究弗林蛋白酶在致癌过程中功能的小鼠模型，促使我们设计了一系列使用转基因小鼠的实验，以促进更好地了解这种重要的 PC 在皮肤癌发病机制中的作用。我们本提案的目标是使用过表达 PC 的转基因小鼠确定弗林蛋白酶单独或与 PACE4 联合促进肿瘤发展的机制。为此设计了两个目标：首先，我们将确定单独的弗林蛋白酶体内靶向过度表达是否会增强紫外线诱导的实验性皮肤癌的发展。我们将使用双基因小鼠研究 PACE4 和弗林蛋白酶在体内同时过度表达，以便确定两种 PC 在皮肤癌发生中可能的合作。将在源自单基因（K5-PACE4和K5-Furin）以及双基因小鼠（K5-PACE4/Furin）的角质形成细胞培养物中评估底物特异性。这些实验将阐明弗林蛋白酶促增殖作用的重要性以及其促侵袭作用与PACE4的主要促侵袭功能是否相同或不同。此外，这一具体目标将确定两种 PC 的组合是否对这些生物过程中的任何一个或两个都有增强作用。其次，我们还将确定 PC 抑制是否会阻止转基因模型中的肿瘤生长和发育。将单基因 PC 转基因小鼠以及双转基因 K5-Furin/PACE4 与表达竞争性 PC 肽抑制剂 cDNA 或 PC 生理抑制剂 cDNA 的 K5-PDX 和 K5-PC-Propeptide 转基因小鼠杂交，我们将确定是否PCs 的抑制改变了由紫外线致癌引起的皮肤肿瘤的形成。公共健康相关性：该应用程序将使用现代实验动物模型来剖析前蛋白转化酶（激活相关癌症相关生物分子的酶）在鳞状细胞癌（人类皮肤最常见的癌症之一）的早期形成和生长过程中的功能。拟议的研究将有助于确定癌症起源的机制，这对于癌症的早期诊断和治疗具有重要的实用价值。