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

研究建议摘要 肝细胞癌（HCC）是一种高致死性的恶性肿瘤，是一个世界性的公共卫生问题.的 HCC患者的主要癌症相关死亡原因是转移性肿瘤进展。但是， HCC转移的分子机制还不清楚。此外，没有准确的预测 可以帮助早期检测侵袭性HCC并从而预测临床结果的血浆生物标志物， 指导治疗选择。我们已经开发了一种转移性HCC的本地小鼠模型， MYC和Twist 1基因在小鼠肝脏中的条件性过表达导致肝脏的逐步进展 模拟人HCC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，并确定HCC的蛋白质组学生物标志物。我们的中心假设是 Twist1调节先天免疫系统特异性蛋白质组和糖蛋白质组的变化， 转移性进展在目标1中，我们将对MYC HCC进行蛋白质组学和糖蛋白质组学分析， MYC/Twist 1 HCC的质谱分析。然后我们将采用系统生物学方法， 蛋白质组学数据，并进行基因敲除实验，以获得关于Twist1如何促进 转移在目标2中，我们将鉴定和验证转移性肿瘤的血浆蛋白质组学和糖蛋白质组学生物标志物。 MYC/Twist 1小鼠中与人HCC相关的进展。成功完成这项研究， 通过鉴定预后生物标志物，这将改善治疗分配，具有显著的转化潜力。 候选人职业发展计划 此K08指导奖申请的目的是使Renumathy Dhanasekaran博士能够承担 监督研究和职业发展培训，成为一个独立的医生科学家。她是一个 董事会认证的肝病学家，2015年加入斯坦福大学医学系， 在Dean Felsher博士的指导下进行转化研究培训。她的长期职业目标是建立 一个独立的转化研究计划，以确定HCC的生物标志物。这款K08将为她提供 保护时间和支持需要进行以下培训：（1）接受先进的实践培训 癌症生物学技术（2）获得计算生物学技能，以进行基因组的综合分析 数据;（3）获得在学术医学中取得成功所需的关键人才， 沟通技巧和（4）收集申请NIH R01补助金所需的初步数据，在第4 - 5年， K08奖。候选人有一个优秀的导师，在癌症生物学方面具有丰富的专业知识，并在一个良好的工作- 资助实验室消化科主任也将担任她的共同导师，他本人 致力于她的学业成功。她组建了一个由经验丰富的癌症专家组成的导师团队 生物学，高级蛋白质组学，生物医学信息学和临床肝病学，谁将担任她的导师 咨询委员会。斯坦福大学是一所世界级的机构，在癌症生物学、计算 生物信息学和转化医学，因此是理想的环境，以促进她的学术生涯。

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.