Targeting Ischemia-Induced Dependencies on the Metabolic Stress Response in Hepatocellular Carcinoma Through Image-Guided, Locoregional Therapy

手术切除或肝移植仍然是肝细胞癌患者唯一的治疗选择。 癌（HCC）。然而，只有不到20%的HCC患者适合切除。经动脉 化疗栓塞（TACE）是一种涉及肝动脉的血管内局部栓塞治疗 动脉内灌注化疗剂的栓塞，被认为是治疗 治疗其余80%的不可切除的HCC患者。虽然TACE的生存率已得到证实， 但是，局部复发是常见的，长期生存率很低。此外，只有44%的接受治疗的HCC 在TACE后病理学上显示广泛坏死，表明肿瘤细胞发展为适应性坏死， 代谢应激反应（MSR），使他们的生存下TACE诱导的营养和氧气剥夺。 在初步研究中，我们已经证明HCC细胞可能被预先编程以在TACE诱导的肿瘤细胞中存活。 通过增强MSR的功能，包括自噬和缺氧诱导因子（HIF），可改善缺血。 此外，TACE诱导的局部缺血导致存活的HCC细胞静止，并进一步激活细胞内的凋亡。 MSR。这些数据表明，TACE提供了一个独特的机会，以限制代谢表型， 以在HCC中生成可确定目标的依赖关系。拟议的项目将建立在这一先前的工作，以：1）进一步 表征靶向MSR途径在TACE样缺血下使HCC细胞存活中的作用， 2)验证靶向这种MSR依赖性的协同治疗策略，以定义一种新的， 更有效的TACE方法。基因编辑将用于检查果糖二磷酸酶1的作用 和尿素循环酶分别影响HCC细胞中HIF和自噬的基础活性。的 HCC细胞存活对每种MSR途径单独和组合的依赖性将是 通过基因编辑和药理学抑制来确定。我们最近开发的，独特的本土 将使用HCC和TACE的大鼠模型来评估TACE诱导的每个MSR途径的重要性， 体内使用MSR的药理学抑制剂。 我们推测，诱导TACE诱导的缺血后存活的HCC细胞的静止使它们 依赖于MSR通路，其可以靶向增强TACE的细胞毒性作用。为了验证这一 假设拟议的项目将追求三个目标：（1）定义有助于 在体外和体内HCC中的分子和细胞应激反应途径激活;（2）功能上 证明存活于严重缺血的HCC细胞对体外MSR诱导的依赖性;和（3） 确定通过单独和同时药理学方法增强TACE诱导的缺血的功效， 体内靶向MSR。重要的是，拟议的项目强调了一种翻译方法， 这些目标的实现将直接影响不可切除HCC患者的治疗， 不治之症

Surgical resection or liver transplantation remain the only curative options for patients with hepatocellular carcinoma (HCC). However, fewer than 20% of patients with HCC are candidates for resection. Transarterial chemoembolization (TACE) is an endovascular locoregional embolotherapy that involves hepatic artery embolization with intra-arterial infusion of a chemotherapeutic agent and is considered the standard of care for treating unresectable HCC in the remaining 80% of patients with this disease. While TACE has a proven survival benefit, local recurrence is common, and long-term survival rates are poor. Moreover, only 44% of treated HCCs demonstrate extensive necrosis on pathology following TACE, indicating that tumor cells develop an adaptive metabolic stress response (MSR) enabling their survival under TACE-induced nutrient and oxygen deprivation. In preliminary studies, we have demonstrated that HCC cells may be pre-programmed to survive TACE-induced ischemia through enhanced function of the MSR, including autophagy and hypoxia-inducible factors (HIFs). Moreover, TACE-induced ischemia results in quiescence in surviving HCC cells and further activation of the MSR. These data demonstrate that TACE offers a unique opportunity to constrain metabolic phenotypes in order to generate targetable dependencies in HCC. The proposed project will build on this prior work to: 1) further characterize the role of targetable MSR pathways in enabling HCC cell survival under TACE-like ischemia and 2) validate a synergistic therapeutic strategy that targets this MSR dependence in order to define a novel, and more effective, approach to TACE. Gene editing will be used to examine the roles of fructose bisphosphatase 1 and urea cycle enzymes in influencing the basal activity of HIFs and autophagy, respectively in HCC cells. The dependence of HCC cell survival on each of the MSR pathways individually and in combination will be determined through gene editing and pharmacologic inhibition. Our recently developed, unique autochthonous rat model of HCC and TACE will be utilized to assess the TACE-induced essentiality of each MSR pathway in vivo using pharmacological inhibitors of the MSR. We hypothesize that the induction of quiescence in HCC cells surviving TACE-induced ischemia renders them dependent on MSR pathways which can be targeted to potentiate the cytotoxic effects of TACE. To test this hypothesis the proposed project will pursue three aims: (1) to define genetic alterations that contribute to molecular and cellular stress response pathway activation in HCC in vitro and in vivo; (2) to functionally demonstrate the dependence of HCC cells surviving severe ischemia on induction of the MSR in vitro; and (3) to determine the efficacy of potentiating TACE-induced ischemia by individual and simultaneous pharmacologic targeting of the MSR in vivo. Importantly, the proposed project emphasizes a translational approach, so that the achievement of these aims will directly inform and influence the treatment of patients with unresectable HCC, an incurable disease.