High Throughput Screening for SCAP Inhibitors as Pancreas Cancer Therapeutics

项目总结 摘要胰腺导管腺癌是美国第四大癌症致死原因。 到2030年，它将成为癌症相关死亡的第二大原因。PDAC通常是 在疾病进程的后期诊断时，治疗选择严重有限，导致总体 确诊后存活时间为12个月，5年存活率为10%。因此，新的药物 预防PDAC原发和转移肿瘤的生长是迫切需要的。 PDAC肿瘤细胞具有极强的增殖性，因此对膜脂的需求很高。 然而，pDAC肿瘤微环境血运不良，导致缺氧和营养有限。 供给。脂肪合成是高度耗氧的，因此肿瘤细胞在低氧环境中是 面临着满足脂类需求的挑战。固醇调节元件结合蛋白(SREBP) 转录因子通过增加胆固醇、脂肪的合成和摄取来维持细胞的脂质供应 酸和甘油三酯。当脂质供应不足时，SREBP被激活，SREBP裂解激活 蛋白(SCAP)是途径活动所必需的。因此，肿瘤应该需要SCAP来适应营养- 限制条件，为膜结构提供脂类。 到目前为止，SREBP通路已经被认为与几种癌症的生长和发展有关， 包括结肠、前列腺、乳房和胶质母细胞瘤。然而，SREBPS在PDAC中的作用并不是 检查过了。使用多个人PDAC细胞系和小鼠肿瘤模型，我们证明了SCAP是 这是PDAC肿瘤发生和生长所必需的，使SCAP成为治疗的新靶点。 尽管在多种癌症中对SREBP途径有既定的要求，但没有有效的化学物质 抑制物存在。为了确定新的候选治疗药物，我们开发了一种强大的高通量检测方法 SCAP抑制剂。在这里，我们以我们的初步研究为基础，确定有效的SCAP化学抑制剂 胰腺癌的治疗。 目的1.通过高通量筛选筛选出SCAP的候选抑制剂。 目的2.验证SCAP抑制剂并进行初步的合成孔径雷达研究。 这些研究的总体目标是确定一系列有效的、化学上易处理的、有效的化合物。 在至少两个正交试验中抑制SCAP，剂量反应超过100倍的浓度范围。 针对癌症脂质代谢的传统方法主要集中在抑制一种单一的酶。 胆固醇、类异戊二烯或脂肪酸合成所需。SREBPs直接调控30多个基因 脂类代谢。因此，抑制SCAP将协同阻断多种合成和摄取途径，并 代表了一种新的靶向癌症脂代谢的方法。

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of death due to cancer in the United States and will become the 2nd leading cause of cancer-related death by 2030. PDAC is typically diagnosed late in the disease process when therapeutic options are severely limited, resulting in an overall survival time of <12 months post-diagnosis and a 5-year survival rate of <10%. Therefore, new drugs that prevent PDAC primary and metastatic tumor growth are desperately needed. PDAC tumor cells are extremely proliferative and thus have a high requisite demand for membrane lipids. However, the PDAC tumor microenvironment is poorly vascularized, leading to hypoxia and limited nutrient supply. Lipid synthesis is highly oxygen-consumptive, so neoplastic cells in a hypoxic environment are challenged with meeting the demand for lipids. The sterol regulatory element-binding protein (SREBP) transcription factors maintain cellular lipid supply by increasing synthesis and uptake of cholesterol, fatty acids, and triglycerides. SREBPs are activated when lipid supply is low, and SREBP cleavage activating protein (SCAP) is required for pathway activity. Thus, tumors should require SCAP to adapt to nutrient- limiting conditions and supply lipid for membrane construction. To date, the SREBP pathway has been implicated in the growth and progression of several cancers, including colon, prostate, breast, and glioblastoma. However, the role of the SREBPs in PDAC has not been examined. Using multiple human PDAC cell lines and mouse tumor models, we demonstrated that SCAP is required for PDAC tumor development and growth, making SCAP a novel target for therapeutics. Despite the established requirement for the SREBP pathway in multiple cancers, no potent chemical inhibitor exists. To identify new candidate therapeutics, we developed a robust high throughput assay for SCAP inhibitors. Here, we build on our preliminary studies to identify potent SCAP chemical inhibitors for the treatment of pancreas cancer. AIM 1. To identify candidate SCAP inhibitors by high throughput screening. AIM 2. To validate SCAP inhibitors and conduct preliminary SAR studies. The overall goal of these studies is to identify a series of potent, chemically tractable, validated compounds that inhibit SCAP in at least two orthogonal assays with a dose-response over 100-fold concentration range. Traditional approaches to target lipid metabolism in cancer have focused on inhibition of a single enzyme required for cholesterol, isoprenoid or fatty acid synthesis. SREBPs directly regulate more than 30 genes of lipid metabolism. Thus, inhibiting SCAP will coordinately block multiple synthesis and uptake pathways and represents a novel approach to targeting lipid metabolism in cancer.