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: 8029552
• 负责人: ANDRES J KLEIN-SZANTO
• 金额: $ 35.12万
• 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/8029552
• 关键词: Biological Process; Biomedical Engineering; Cell Proliferation; Complementary DNA; Development; Early Diagnosis; Early treatment; Enzymes; Epidermis; Experimental Animal Model; Family; Growth; Growth Factor; Growth Factor Receptors; Growth and Development function; Health; 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; 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; 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）：弗林蛋白酶是前蛋白转化酶（PC）家族的蛋白加工酶，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，重点是小鼠皮肤暴露于紫外线后发生的早期和晚期变化。PACE 4是另一种相关的PC，已显示通过激活相关蛋白酶在鼠肿瘤细胞侵袭中起作用。弗林蛋白酶还能够通过激活生长因子和生长因子受体（如IGF-R1）来增强人类癌前和癌细胞中的细胞增殖。缺乏一个小鼠模型来研究弗林蛋白酶在癌变过程中的功能，促使我们设计了一系列的实验，使用转基因小鼠，以促进更好地了解这一突出的PC在皮肤癌的发病机制。我们在本提案中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PACE 4联合促进肿瘤发展的机制。两个目标已被设计为这种效果：首先，我们将确定是否有针对性的体内过度表达弗林蛋白酶单独将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PACE 4和弗林蛋白酶的同时体内过表达，以建立2个PC在皮肤癌发生中的可能合作。将在来源于单基因（K5-PACE 4和K5-Furin）以及双基因小鼠（K5-PACE 4/Furin）的角质形成细胞培养物中评价底物特异性。这些实验将阐明弗林蛋白酶促增殖作用的意义，以及其促侵袭作用与PACE 4的主要促侵袭功能是否相同或不同。此外，这一特定目标将确定两种PC的组合是否对这些生物过程中的任何一种或两种具有增强作用。其次，我们还将确定PC抑制是否会阻断转基因模型中的肿瘤生长和发展。将单基因PC转基因小鼠以及双转基因K5-Furin/PACE 4与表达竞争性PC肽抑制剂或PC生理抑制剂的cDNA的K5-PDX和K5-PC-前肽转基因小鼠杂交，我们将确定PC的抑制是否改变UV致癌诱导的皮肤肿瘤的形成。公共卫生关系：该应用将使用现代实验动物模型来解剖前蛋白转化酶的功能，前蛋白转化酶是在鳞状细胞癌（人类皮肤最常见的癌症之一）的早期形成和生长期间激活相关癌症相关生物分子的酶。这些研究将有助于确定癌症起源的机制，这对癌症的早期诊断和治疗具有重要的实用价值。