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

翻译后摘要：由于转移性肝细胞癌（HCC）的死亡继续安装由于低成功率， 临床干预。靶向癌症治疗消除了所有恶性肿瘤引发的干细胞样细胞（ TICs）至 防止复发和转移。HCC的发生与酒精和/或HCV感染有关， 常见于基因畸变的患者， SWI/SNF复合体。我们基于全基因组CRISPR敲除的合成致死基因筛选 我们的人源化小鼠模型的ARID 1A突变细胞中的（GeCKO）-慢病毒文库鉴定了组分 (EZH2和EED）。我们证明了多能性转录 NANOG因子代谢重编程TIC，通过抑制氧化应激促进自我更新。 磷酸化（OXPHOS）和脂肪酸氧化活化（FAO）。我们将酒精相关的 人肝癌和酒精喂养的HCV NS 5A转基因（Tg）小鼠在肝癌发展中的关键作用 NANOG作为核心干细胞因子，在TLR 4活化后作为发生和分化所需的关键组分， 维护TIC。 这些发现支持了我们的假设，即酒精介导的NANOG诱导和ARID 1A突变 在酒精相关性HCC中协同产生耐药TIC。因此，针对 ARID1A，a 在染色质重塑 NANOG-PRC 2综合体和粮农组织应消除TIC亚群。我们提出三个具体目标。在 目的1，我们将测试NANOG与PRC 2复合物是否抑制OXPHOS并产生索拉非尼抗性TIC， 突变ARID 1A在酒精介导的人HCC中的表达。在目标2中，我们将研究抑制FAO如何抑制 自我更新和促进索拉非尼耐药突变ARID 1A肿瘤的细胞凋亡。在目标3中，我们将测试 索拉非尼加PRC 2/FAO抑制的联合治疗诱导代谢重编程和细胞凋亡， HCC。我们将测试靶向合成致死基因（EZH 2）是否通过使用以下方法消除具有ARID 1A突变的TIC： 酒精西方饮食喂养和/或HCV感染的FRG-hu-Hep/HSC人源化小鼠和患者来源的 异种移植 总之，我们的基因型HCC模型将简化疾病建模/毒性评价的临床试验 通过提供“在人源化小鼠中进行临床试验”。PDX和人源化小鼠系统的使用将提供 这是一条通往范式转变的个性化医疗的道路，以实现准确的合成致命性靶向策略。及时 根除由长期酒精暴露引起的耐药TIC， 有益且具有成本效益， 患有不治之症的病人。 这项建议将建立一个个性化的治疗 在muco-ARID 1A HCC中靶向合成致死途径（PRC 2）-NANOG复合物和FAO的策略。一 成功的结果将导致范式的转变，从而促进个性化的未来发展 针对酒精相关HCC中出现的NANOG+ TIC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.