NANOG-positive cancer stem cells in liver oncogenesis induced by alcohol and HCV

摘要：由于转移性肝细胞癌的治疗成功率较低，导致的死亡人数持续上升。 临床干预。靶向癌症治疗消除了所有启动恶性肿瘤的干细胞( TICS)至 防止复发和转移。肝细胞癌的发生与酒精和/或丙型肝炎病毒感染有关 常见于成分基因异常患者 复杂的SWI/SNF。基于CRISPR基因敲除的全基因组合成致死基因筛选 (壁虎)慢病毒文库在我们人源化小鼠模型的ARID1A突变细胞中鉴定出成分 (EZH2和EED)的多梳抑制复合体2(PRC2)。我们证明了多能性转录 因子NANOG通过抑制氧化作用代谢重新编程TIC以促进自我更新 磷酸化(OXPHOS)和激活脂肪酸氧化(FAO)。我们描述了酒精相关的 人肝细胞癌和酒精喂养的丙型肝炎病毒NS5A转基因(TG)小鼠在肝癌发展中的关键作用 Nanog是继TLR4激活之后的核心干细胞因子，是发生和发展所需的关键成分 抽搐的维护。 这些发现支持我们的假设，酒精介导的NANOG诱导和ARID1A突变 在酒精相关的肝细胞癌中协同产生化疗耐药药物。因此，目标是 ARID1a，a 染色质重塑 Nanog-PRC2复合体和粮农组织应消除TIC亚群。我们提出了三个具体目标。在……里面 目的1，我们将测试具有PRC2复合体的NANOG是否抑制OXPHOS并产生对索拉非尼耐药的TICE 酒精介导的人肝癌细胞中的突变ARID1A。在目标2中，我们将研究粮农组织的抑制如何抑制 对索拉非尼耐药突变ARID1A肿瘤的自我更新和促进细胞凋亡。在目标3中，我们将测试 索拉非尼联合PRC2/FAO抑制诱导细胞代谢重编程和细胞凋亡 肝细胞癌。我们将测试靶向合成致死基因(EZH2)是否通过使用以下方法消除具有ARID1A突变的TICS 酒精西方饮食喂养和/或感染丙型肝炎病毒的FRG-Hu-Hep/HSC人源化小鼠和患者来源 异种移植。 总之，我们的基因分型肝细胞癌模型将简化用于疾病建模/毒性评估的临床试验 通过提供“在人性化小鼠身上进行的临床试验”。使用PDX和人性化的鼠标系统将提供 针对精确的合成致命性靶向策略的范式转换个性化医学的路径。适时的 根除因长期接触酒精而产生的耐药药物将是 有益且经济实惠 患有不治之症的病人。 这项提议将建立一种个性化的治疗方法 针对突变-ARID1A型肝癌的合成致死途径(PRC2)-NANOG复合体和粮农组织的战略。一个 成功的结果将导致范式的转变，从而促进个性化未来的发展 针对酒精相关性肝癌中出现的NANOG抽搐的治疗策略。

Abstract: Deaths due to metastatic hepatocellular carcinoma (HCC) continue to mount due to a low success of clinical intervention. Targeted cancer therapy eliminates all malignant tumor-initiating stem-like cells ( TICs) to prevent relapse and metastasis. Development of HCC is associated with alcohol and/or HCV infection and frequently observed in patients with gene aberrations in component complex SWI/SNF. Our synthetic lethal gene screening of genome-wide CRISPR-based knockout (GeCKO)-lentivirus library in ARID1A-mutant cells of our humanized mouse models identified components (EZH2 and EED) of polycomb suppressor complex 2 (PRC2). We demonstrated that pluripotency transcription factor NANOG metabolically reprograms TICs to promote self-renewal via inhibition of oxidative phosphorylation (OXPHOS) and activation of fatty acid oxidation (FAO). We characterized alcohol-associated human HCCs and alcohol-fed, HCV NS5A transgenic (Tg) mice for HCC development and a critical role of NANOG as a core stem cell factor following TLR4 activation as a crucial component needed for genesis and maintenance of TICs. These findings support our hypothesis that alcohol-mediated NANOG induction and ARID1A mutations cooperatively generate chemoresistant TICs in alcohol-associated HCC. Therefore, targeting ARID1A, a of chromatin remodeling NANOG-PRC2 complex and FAO should eliminate the TIC subpopulation. We propose three specific aims. In Aim 1, we will test if NANOG with PRC2 complex inhibits OXPHOS and generates sorafenib-resistant TICs with mutant ARID1A in alcohol mediated human HCCs. In Aim 2, we will examine how inhibition of FAO inhibit self-renewal and promote apoprtosis in sorafenib-resistant mutant ARID1A tumors. In Aim 3, we will test if combination therapy of sorafenib plus PRC2/FAO inhibition induces metabolic reprogramming and apoptosis in HCC. We will test if targeting synthetic lethality genes (EZH2) eliminates TICs with ARID1A mutations by use of alcohol Western diet-fed and/or HCV-infected FRG-hu-Hep/HSC humanized mice and patient-derived xenograft. In summary our genotyped HCC model will streamline clinical trials for disease modeling/toxicity evaluation by offering “a clinical trial in a humanized mouse”. Use of PDX and humanized mouse systems will provide a path to paradigm-shifting personalized medicine for an accurate synthetic lethality targeting strategy. The timely eradication of chemoresistant TICs arising from chronic alcohol exposure will be beneficial and cost-effective for patients suffering from an otherwise incurable disease. This proposal will establish a personalized therapy strategy targeting a synthetic lethal pathway (PRC2)-NANOG complex and FAO in mutant-ARID1A HCCs. A successful outcome will lead to a paradigm shift, thus facilitating the future development of personalized treatment strategies targeted towards NANOG+ TICs appearing in alcohol-related HCC.