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 癌症(HCC)。然而，只有不到20%的肝细胞癌患者适合切除。经动脉 化疗栓塞术(TACE)是一种涉及肝动脉的血管内局部栓塞术 动脉内注入化疗药物的栓塞，被认为是治疗的标准 治疗剩余80%不能切除的肝细胞癌患者。虽然TACE的存活率得到了证实 受益，局部复发很常见，长期存活率很低。此外，只有44%的经过处理的氟氯烃 TACE后病理上显示广泛的坏死，表明肿瘤细胞发展成适应性 代谢应激反应(MSR)使它们能够在TACE诱导的营养和缺氧剥夺下存活。 在初步研究中，我们已经证明，肝癌细胞可能是预先编程的，能够在TACE诱导下存活 缺血通过增强MSR的功能，包括自噬和低氧诱导因子(HIF)。 此外，TACE诱导的缺血导致存活的肝癌细胞停滞，并进一步激活 MSR。这些数据表明，TACE提供了一个独特的机会来限制代谢表型的顺序 在肝细胞癌中生成目标依赖关系。拟议的项目将在这一先前工作的基础上：1)进一步 表征靶向MSR通路在使肝癌细胞在TACE样缺血和死亡中存活的作用 2)验证针对这种MSR依赖的协同治疗策略，以定义一种新的 更有效的TACE方法。基因编辑将被用来研究果糖二磷酸酶1的作用 和尿素循环酶分别影响肝癌细胞HIFs和自噬的基础活性。这个 肝癌细胞的存活依赖于每个MSR通路的单独和组合将是 通过基因编辑和药物抑制来确定。我们最近开发的独一无二的原产地 将利用大鼠肝癌模型和TACE来评估TACE诱导的每一条MSR途径在 体内使用MSR的药理抑制剂。 我们假设，在TACE诱导的缺血中存活的肝细胞癌细胞的静止诱导使它们 依赖于MSR通路，可靶向增强TACE的细胞毒效应。为了测试这一点 假设拟议的项目将追求三个目标：(1)定义有助于 分子和细胞应激反应通路在肝癌体内和体外的激活；(2)功能性 证明肝癌细胞在严重缺血中存活对体外诱导MSR的依赖性；以及(3) 单药和同时用药对TACE诱导的脑缺血的增强作用 体内靶向MSR。重要的是，拟议的项目强调翻译方法，因此 这些目标的实现将直接通知和影响不能切除的肝癌患者的治疗。 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.