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.

Public Health Relevance

The proposed studies address the pathogenic development of hepatocellular carcinoma (HCC), an important public health problem mechanisms underlying the genesis of TIC-reprograming factors and pathways that continues to evade our best therapeutic efforts. We will examine tumor initiating stem-like cells (TIC) and elucidate the with a major emphasis on as a major reprogramming factor for oncogenesis and therapy resistance. Completion of the proposed aims will lead to ground-breaking personalized medicine with drug therapy targeting TICs and development of an ARID1A mutant HCC to streamline clinical trials for disease modeling/toxicity evaluation to achieve ?a clinical trial in human livers and human immune systems in mice? system.

National Institute of Health (NIH)
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Research Project (R01)
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Hepatobiliary Pathophysiology Study Section (HBPP)
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Murray, Gary
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University of Southern California
Schools of Medicine
Los Angeles
United States
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