Reverse-engineering precision liver cancer chemoprevention

确定临床上相关的癌症化学预防靶点一直是具有挑战性的。我们的多学科 Team(精密肝癌预防联盟)将采用创新的逆向工程方法， 从对长期临床随访的存档临床标本进行转录组分析开始，然后 转移到只对临床上经过充分验证的靶点进行多重实验验证，以阐明 临床应用成功的可能性最高的化学预防目标。通过这种方法，我们 可以成功识别肝癌风险特征和化学预防措施，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，我们的目的是阐明受非传染性疾病影响的肝脏致癌环境潜在的分子失调。 酒精性脂肪性肝炎(NASH)，上升最快的肝癌病因，作为改进化学预防的线索 靶标、药物和生物标记物分析，有望实现个性化的患者管理和 更具成本效益的肝癌化学预防临床试验，并导致革命性的改善 患者预后与建立新范式，逆向工程精准预防癌症。 目的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通过计算优先排序，并与LPA一起进行筛选 肝癌风险信号诱导细胞系统中用于基因信号逆转的途径抑制物库。为 在屏幕上选择的化合物，作用机制将通过功能的得失来询问 细胞系统中的评估。 目的2.候选肝癌化学预防药物的功能验证。活体肝癌 候选药物的化学预防作用将在饮食诱导的纤维化/致癌大鼠中得到验证 模仿全球人类肝硬变转录组的模型。代理人的人类相关性将在以下方面进行评估 临床肝纤维化组织器官型体外培养(n=30)。确定目标细胞类型(S)和 作用机制为药剂，主要的肝细胞类型将从大鼠体内分离出来，并转录组 将进行简档分析以评估肝癌风险签名成员基因，推断的目标基因 化合物，以及相关的分子途径。人类纤维化/肝硬变的纳什肝脏将被类似地描述为 细胞类型特异性转录调控失调，以验证调节基因在人类中是否相关。 目的3.发展基于组织和血清的肝癌风险生物标记物分析。以肝组织为基础 癌症风险标记将在临床适用的组织-(NanoString)和血清-(Luminex)中实施 基于分析，并在200人的队列中评估技术有效性和预测未来癌症风险的能力 NASH患者的配对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.