Functional Proteomic Analysis and Biomarker Identification in a Novel Mouse Model of Metastatic Hepatocellular Carcinoma (HCC)

研究提案摘要 肝细胞癌是一种高度致命性的恶性肿瘤，是一个世界性的公共卫生问题。这个 肝细胞癌患者的主要癌症相关死亡原因是转移的肿瘤进展。但准确的是 肝细胞癌转移的分子机制还不是很清楚。此外，也没有准确的预测 血浆生物标记物有助于侵袭性肝癌的早期发现，从而预测临床预后或 指导治疗选择。我们已经建立了一种转移性肝癌的自体小鼠模型 MYC和Twist1基因在小鼠肝脏的条件性过表达导致肝脏的渐进性进展 模拟人类肝细胞癌的癌症。本研究建议使用这种新的转移性肝癌小鼠模型来探索 肿瘤进展的机制和确定肝细胞癌的蛋白质组生物标志物。我们的中心假设是 Twist1调节先天免疫系统特异的蛋白质组和糖蛋白质组变化，从而促进 转移的进展。在目标1中，我们将进行MYC肝细胞癌和糖蛋白组学分析。 MYC/Twist1肝癌的质谱学分析。然后，我们将采用大规模的系统生物学方法。 蛋白质组学数据和执行基因敲除实验，以获得Twist1如何促进 转移。在目标2中，我们将识别和验证转移的血浆蛋白质组和糖蛋白质组生物标记物。 与人类肝细胞癌相关的MYC/Twist1小鼠的进展。这项研究的成功完成已 通过识别将改善治疗分配的预后生物标记物，具有显著的翻译潜力。 应聘者职业发展计划 K08导师奖申请的目标是使Renumathy Dhanasekaran博士能够进行 指导研究和职业发展培训，成为一名独立的内科科学家。她是个 董事会认证肝病专家，2015年加入斯坦福大学医学系，目前正在攻读高级 在Dean Felsher博士的指导下进行翻译研究培训。她的长期职业目标是建立 一项独立的翻译研究计划，以确定肝细胞癌的生物标记物。这款K08将为她提供 受保护的时间和开展以下培训所需的支持：(1)接受高级实践培训 癌症生物学中的技术(2)获得计算生物学中的技能以执行基因组的综合分析 数据；(3)获取在学术医学领域取得成功所需的关键人才，如格兰特精神；以及 沟通技能和(4)收集在第四至第五年申请NIH R01拨款所需的初步数据 K08奖。候选人有一位出色的导师，在癌症生物学方面拥有丰富的专业知识，并在一口井中工作- 资助的实验室。消化内科主任也将担任她的联合导师，他本人 致力于她的学术成就。她已经组建了一支经验丰富、具有癌症专业知识的导师团队 生物学、高级蛋白质组学、生物医学信息学和临床肝病学，她将担任她的导师 咨询委员会。斯坦福大学是一所世界级的学府，在癌症生物学、计算机科学方面具有特别的优势。 生物信息学和转化医学，因此是推进她的学术生涯的理想环境。

Research Proposal Summary Hepatocellular carcinoma (HCC) is a highly lethal malignancy, and is a worldwide public health problem. The main cancer-related cause of death in patients with HCC is metastatic tumor progression. But the precise molecular mechanisms of HCC metastasis are not well understood. Also, there are no accurate prognostic plasma biomarkers which can help in early detection of aggressive HCC and thereby predict clinical outcome or guide treatment selection. We have developed an autochthonous mouse model of metastatic HCC where conditional overexpression of MYC and Twist1 genes in the mouse liver leads to step-wise progression of liver cancer simulating human HCC. This study proposes to use this novel mouse model of metastatic HCC to explore the mechanisms of tumor progression and identify proteomic biomarkers for HCC. Our central HYPOTHESIS is that Twist1 modulates innate immune system specific proteomic and glycoproteomic changes which promote metastatic progression. In Aim 1 we will perform proteomic and glycoproteomic analyses of MYC HCC and MYC/Twist1 HCC using mass spectrometry. We will then undertake a systems biology approach to large scale proteomics data and perform gene knockout experiments to gain mechanistic insights into how Twist1 promotes metastasis. In Aim 2 we will identify and validate plasma proteomic and glycoproteomic biomarkers of metastatic progression in the MYC/Twist1 mice that are relevant to human HCC. Successful completion of this study has significant translational potential by identifying prognostic biomarkers which will improve treatment allocation. Candidate Career Development Plan The objective of this K08 Mentored Award application is to enable Dr. Renumathy Dhanasekaran to undertake supervised research and career development training to become an independent physician scientist. She is a board certified Hepatologist who joined the Department of Medicine at Stanford in 2015 and is pursuing advanced translational research training under the mentorship of Dr. Dean Felsher. Her long term career goal is to build an independent translational research program to identify biomarkers for HCC. This K08 will provide her with the protected time and support needed to undertake the following training: (1) receive hands-on training in advanced techniques in cancer biology (2) acquire skills in computational biology to perform integrated analyses of genomic data; (3) acquire critical talents required for success in academic medicine like grantsmanship, and communication skills and (4) gather preliminary data required to apply for a NIH R01 grant during year 4-5 of the K08 award. The candidate has an excellent mentor with vast expertise in cancer biology and is working in a well- funded lab. The Chief of the Division of Gastroenterology, will also serve as her co-mentor and he is personally committed to her academic success. She has assembled a team of seasoned mentors with expertise in cancer biology, advanced proteomics, biomedical informatics and clinical hepatology, who will serve on her mentorship advisory committee. Stanford is a world-class institution with particular strength in cancer biology, computational bioinformatics and translational medicine, and hence is the ideal environment to advance her academic career.