Evaluation of voltage-gated calcium ion channels as a therapeutic target in intrahepatic cholangiocarcinoma

项目摘要/摘要 肝内胆管细胞癌(ICC)是一种侵袭性的原发性肝癌，总生存率的中位数为 11.7个月。大多数患者患有不能切除的疾病，显示出显著的需要 改进了系统疗法。在这里，我们使用CRISPR/Cas9筛查来确定ICC生存所必需的基因 可以作为新的治疗靶点。我们证明了编码CACNA1a和CACNA1G的 两个电压门控钙离子通道(VGCC)的主要成孔亚基是 在五种ICC细胞系中的重要性。我们假设电压门控离子通道的异常表达是 对ICC生存至关重要，并建议现有的FDA批准的钙通道阻滞剂可以 重新用于治疗国际刑事法院。我们的初步数据表明，这两个基因不仅对ICC是必不可少的 而且，抗高血压药和抗精神病药物在降低ICC方面也可能有效 体外存活能力。为了验证我们的中心假设，我们提出了以下目标：1)确定VGCC的效果 体外对ICC细胞的基因敲除和药物阻断，2)探讨其作用机制。 这些钙通道的阻断降低了ICC的活性，3)决定了钙通道 阻滞剂可以单独使用或在体内和新患者中增强化疗的效果- 派生模型。

Project Summary/Abstract Intrahepatic cholangiocarcinoma (ICC) is an aggressive primary liver cancer with a median overall survival of 11.7 months. The majority of patients present with unresectable disease, demonstrating a significant need for improved systemic therapies. Here, we use CRISPR/Cas9 screens to identify genes essential to ICC survival that could serve as novel therapeutic targets. We demonstrate that CACNA1A and CACNA1G, which encode the main pore-forming subunits of two voltage-gated calcium ion channels (VGCCs), are two genes of essentiality in five ICC cell lines. We hypothesize that aberrant expression of voltage-gated ion channels is essential to ICC survival and propose that existing FDA-approved calcium channel blockers could be repurposed to treat ICC. Our preliminary data demonstrate that not only are these two genes essential to ICC proliferation, but also that both antihypertensives and antipsychotics may be effective in decreasing ICC viability in vitro. To test our central hypothesis, we propose the following aims: 1) determine the effect of VGCC genetic knockdown and pharmacologic blockade on ICC cells in vitro, 2) investigate the mechanisms by which blockade of these calcium channels decreases ICC viability, and 3) determine whether calcium channel blockers can be used individually or to augment the effects of chemotherapy both in vivo and in novel patient- derived models.