Reverse-engineering precision liver cancer chemoprevention

临床相关癌症化学预防靶点的鉴定一直具有挑战性。我们的多学科 团队（精准肝癌预防联盟）将采用创新的逆向工程方法， 从长期临床随访的存档临床标本的转录组分析开始，然后 移动到只有临床验证良好的目标的多重实验验证，以阐明 化学预防靶点具有最高的成功临床应用的可能性。通过这种方法，我们 可以成功地识别肝癌风险特征和化学预防药物，从而进行临床试验 （NCT02273362）。为了实现我们建立临床适用的化学预防策略的长期目标， 在这里，我们的目的是阐明在受非肿瘤性肝损伤影响的肝脏中潜在的致癌环境的分子失调， 酒精性脂肪性肝炎（NASH）是增长最快的肝癌病因，作为精细化学预防的线索 靶点、药物和生物标志物检测，有望实现个性化患者管理， 更具有成本效益的肝癌化学预防临床试验，并导致革命性的改善， 患者预后和建立一个新的范例，逆向工程精确预防癌症。 目标1。肝癌化学预防药物的计算机靶向筛选。候选肝脏 癌症化学预防化合物将在计算上优先考虑，并与LPA一起筛选 在肝癌风险信号诱导细胞系统中的通路抑制剂文库中进行基因信号逆转。为 筛选中的选定化合物，将通过功能获得或丧失来询问作用机制 在细胞系统中进行评估。 目标二。候选肝癌化学预防剂的功能验证。体内肝癌 将在饮食诱导的纤维化/致癌大鼠中验证候选药剂的化学预防作用 模拟全球人类肝硬化转录组的模型。将在以下文件中评价药物的人体相关性： 临床纤维化肝组织的器官型离体培养（n=30）。确定靶细胞类型， 为了确定药物的作用机制，将从大鼠中分离主要肝细胞类型， 将进行谱分析以评估肝癌风险特征成员基因，即肝癌风险特征成员基因的推断靶基因。 化合物和相关的分子途径。人类纤维化/硬化NASH肝脏将类似地进行分析， 细胞类型特异性转录组失调，以验证调节的基因在人类中是相关的。 目标3。开发基于组织和血清的肝癌风险生物标志物测定。基于肝脏组织 将在临床适用组织（NanoString）和血清（Luminex）中实施癌症风险特征 基于分析，并评估技术有效性和预测200人队列中未来癌症风险的能力 NASH患者配对的血清和肝组织标本以及完成的长期临床研究 观察癌症的发展。

Identification of clinically relevant cancer chemoprevention targets has been challenging. Our multidisciplinary team (Precision Liver Cancer Prevention Consortium) will employ an innovative reverse-engineering approach, starting from transcriptome analysis of archived clinical specimens with long-term clinical follow-up, then moving to multifold experimental verification of only clinically well-validated targets, to elucidate chemoprevention targets with the highest likelihood of successful clinical application. With this approach, we could successfully identify liver cancer risk signatures and chemoprevention dugs, leading to a clinical trial (NCT02273362). To achieve our long-term goal of establishing clinically applicable chemoprevention strategies, here we aim to elucidate molecular dysregulation underlying carcinogenic milieu in livers affected with non- alcoholic steatohepatitis (NASH), the fastest rising liver cancer etiology, as clues to refined chemoprevention targets, drugs, and biomarker assays, which are expected to enable personalized patient management and more cost-effective liver cancer chemoprevention clinical trials, and lead to revolutionary improvement of patient prognosis and establishment of a new paradigm, reverse-engineering precision cancer prevention. Aim 1. Computationally-targeted screening of liver cancer chemoprevention agents. Candidate liver cancer chemopreventive compounds will be computationally prioritized, and screened together with LPA pathway inhibitor library in liver cancer risk signature-inducible cell system for the gene signature reversal. For selected compounds in the screen, mechanisms of action will be interrogated by gain- or loss-of-function assessment in the cell system. Aim 2. Functional validation of candidate liver cancer chemoprevention agents. In vivo liver cancer chemopreventive effect of the candidate agents will be validated in a diet-induced fibrotic/carcinogenic rat model mimicking global human cirrhosis transcriptome. Human relevance of the agents will be evaluated in organotypic ex vivo culture of clinical fibrotic liver tissues (n=30). To determine target cell type(s) and mechanisms of action for the agents, major hepatic cell types will be isolated from the rats, and transcriptome profiling will be performed to assess liver cancer risk signature member genes, the inferred target genes for the compounds, and related molecular pathways. Human fibrotic/cirrhotic NASH livers will be similarly profiled for cell type-specific transcriptomic dysregulation to verify that the modulated genes are relevant in human. Aim 3. Development of tissue- and serum-based liver cancer risk biomarker assays. Liver tissue-based cancer risk signatures will be implemented in clinically applicable tissue- (NanoString) and serum- (Luminex) based assays, and evaluated for technical validity and capability to predict future cancer risk in a cohort of 200 NASH patients with paired serum and liver tissue specimens as well as completed long-term clinical observation for cancer development.