Targeting of mTOR Activity for Chemoprevention of Solar UV-induced Skin Cancer

项目总结（见说明）： 本项目的假设是，P1 -3激酶-Akt-mTOR和AMP活化蛋白激酶（即AMPK）-mTOR信号通路是开发局部化学预防剂的靶点，通过阻断遗传改变和太阳紫外线引发的角质形成细胞的生长或诱导其死亡来抑制太阳紫外线诱导的非黑素瘤皮肤癌。具体目标1：利用转基因小鼠 确定Akt-1和mTOR在太阳紫外线诱导的小鼠皮肤癌发生中的功能作用。实验方法是开发表皮特异性诱导的Akt-1和mTOR的条件性敲除小鼠，并使用这些小鼠来验证这些分子靶标。这些小鼠将被给予项目2， 研究Akt/mTOR和Fyn/RSK 2通路之间的串扰。具体目标2：使用转基因小鼠来确定AMPK在太阳紫外线诱导的小鼠皮肤癌变中的功能作用。实验方法是开发其中组成型活性AMPK在表皮中特异性表达的转基因小鼠，并使用这些小鼠来验证分子靶标。具体目标3：单独或联合使用Akt和mTOR抑制剂以及AMPK激活剂，以确定这些激酶的药理学调节是否抑制太阳紫外线诱导的mTOR信号传导、细胞增殖、抗凋亡信号传导和皮肤致癌作用。使用的Akt抑制剂是PHT-427。的 mTOR抑制剂是雷帕霉素，AMPK激活剂是蛋氨酸，与项目2合作，我们将测试靶向Akt/mTOR和Fyn/RSK 2信号通路的药物组合具体目标4：交叉验证Akt、mTOR和AMPK作为太阳紫外线诱导皮肤化学预防的分子靶点。 使用组织微阵列和反相蛋白质微阵列对小鼠和人类皮肤之间的癌症进行比较。 这一目标将与项目3合作进行，并将涉及受试者的太阳光照射和用于研究靶信号通路激活的打孔活检。在本临床前项目中显示出化学预防活性的成功先导药物或药物组合将由项目3在临床中进行测试。 这个高度互动和临床转化的研究计划项目的重点是在创新的小鼠模型（项目1和2）中成功进行靶向化学预防剂的临床前测试，然后在风险人群中设计和实施临床试验（项目3）。关于决策树选择过程以及项目和核心之间的交互的详细描述可以在资源格式页面上找到。

PROJECT SUMMARY (See instructions): The hypothesis in this Project is that the Pl-3 kinase-Akt-mTOR and the AMP-activated protein kinase (i.e. AMPK)-mTOR signaling pathways are targets forthe development of topical chemoprevention agents to inhibit solar UV induced non-melanoma skin cancer by blocking the growth of or inducing the death of genetically altered and solar UV-initiated keratinocytes. Specific Aim 1: To use genetically modified mice to determine the functional roles of Akt-1 and mTOR in solar UV-induced mouse skin carcinogenesis. The experimental approach is to develop epidermis specific inducible, conditional knockout mice for Akt-1 and mTOR and use these mice to validate these molecular targets. These mice will be given to Project 2 to study cross talk between the Akt/mTOR and Fyn/RSK2 pathways. Specific Aim 2: To use genetically modified mice to determine the functional role of AMPK in solar UV-induced mouse skin carcinogenesis. The experimental approach Is to develop transgenic mice where constitutively active AMPK is specifically expressed in the epidermis and use these mice to validate the molecular target. Specific Aim 3: To utilize inhibitors of Akt and mTOR as well as AMPK activators independently or in combination to determine whether pharmacological modulation of these kinases inhibits solar UV-induced mTOR signaling, cell proliferation, anti-apoptotic signaling and skin carcinogenesis. The Akt inhibitor to be used is PHT-427. The mTOR Inhibitor is rapamycin and the AMPK activator is Metformin, In collaboration with Project 2 we will test combinations of drugs targeting the Akt/mTOR and Fyn/RSK2 signaling pathways Specific Aim 4: To crossvalidate Akt, mTOR and AMPK as molecular targets for the chemoprevention of solar UV-induced skin cancer between mouse and human skin using tissue microarrays and reverse phase protein microarrays. This aim will be carried out in collaboration with Project 3 and will involve solar light-irradiation of subjects and punch biopsies to be used for studying the activation of the target signaling pathways. Successful lead drugs or drug combinations that show chemoprevention activity in this preclinical Project will be tested in the clinic by Project 3. This highly interactive and clinically translational research program project focuses on the successful preclinical testing of targeted chemoprevention agents in innovative mouse models (Projects 1 and 2) followed by the design and implementation of clinical trials in at risk human populations (Project 3). Detailed descriptions of the decision-tree selection process as well as the interactions between Projects and Cores are found on the Resources Format Page.