A novel regimen to target both pancreatic cancer K-ras and antiapoptotic proteins

一种针对胰腺癌 K-ras 和抗凋亡蛋白的新疗法

• 批准号: 8616128
• 负责人: Fengzhi Li
• 金额: $ 8.49万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8616128
• 关键词: Animal Model; Apoptosis; Apoptosis Inhibitor; Area; BCL2 gene; BIRC4 gene; Cancer Cell Growth; Cancer Patient; Cancer cell line; Cell Line; Cell Survival; Cells; Chemicals; Clinical; Colon; Comprehensive Cancer Center; Development; Disease; Dose; Early Diagnosis; Ensure; Environment; Epithelial; Epithelial Cells; Exhibits; Family; Future; Genetic; Goals; Head and neck structure; Human; Induction of Apoptosis; K-ras Gene; K-ras Oncogene; Lead; Malignant Neoplasms; Malignant neoplasm of pancreas; Medicinal Plants; Modeling; Mus; Mutate; Mutation; Nature; Neoplasm Metastasis; Normal Cell; Oncogenic; Ovarian; Pancreatic duct; Pathway interactions; Patients; Pilot Projects; Proteins; RNA Interference; Regimen; Relapse; Reporting; Research Personnel; Resistance; Severe Combined Immunodeficiency; Side; Signal Pathway; Survival Rate; System; Testing; Tissues; Toxic effect; Treatment Protocols; Tumor Volume; Xenograft Model; Xenograft procedure; cancer cell; cancer type; chemical reaction; design; effective therapy; experience; gain of function; gain of function mutation; gemcitabine; inhibitor/antagonist; innovation; killings; leukemia; mouse model; mutant; novel; pancreatic cancer cells; pancreatic neoplasm; public health relevance; screening; survivin; treatment duration; tumor; tumor growth; tumor xenograft

Abstract The goal of this R03 pilot project is to demonstrate a proof of concept for an innovative combination treatment regimen designed to target pancreatic cancer with K-ras mutations and aberrant expression of one or more antiapoptotic proteins. K-ras oncogenic mutations and aberrant expression of major antiapoptotic proteins (e.g. survivin, Mcl-1, XIAP, cIAP2) in pancreatic cancer heavily contribute to pancreatic cancer development and aggressiveness (treatment resistance, metastasis, and relapse). Gain-of-K-ras-function mutations is observed in >90% of pancreatic cancer patients. Genetic silencing of mutated K-ras induces apoptosis and inhibits pancreatic cancer cell growth, invasiveness, malignant tumor formation, and xenograft tumor growth. However, there are no effective targeted therapies available for pancreatic cancer K-ras mutations. Using a novel screening approach with K-ras mutant cells versus normal cells, fifteen chemical constituents from the medicinal plant Amoora rohituka were identified, and over 50 derivatives were generated using semi-synthetic approaches from the 15 hits. These compounds were then rescreened using K-ras mutant cells versus normal cells. AMR-Me and AMR-MeOAc were identified as the most potent compounds selectively against the K-ras mutant cells. Our previous studies indicated that AMR-Me targets the K-ras pathway, and that 3 mg/kg daily for 28-day treatment of mice shows no clear toxicity, while it extends leukemia mouse survival. We plan to combine AMR-MeOAc (best selectivity) with our novel compound, FL118, which selectively inhibits survivin, Mcl-1, XIAP, and cIAP2, for testing this novel combinational-targeted treatment regimen. It has previously been shown that combination of K-ras silencing with gemcitabine dramatically reduces tumor volumes in mice compared with either single agent alone. Therefore, we hypothesize that inhibition of both mutated K-ras and the major antiapoptotic proteins with the novel agents AMR-MeOAc and FL118 would lead to a strikingly enhanced induction of apoptosis and inhibition of pancreatic cancer cell and tumor growth than the inhibition from either agent alone. The following three specific aims are proposed in this project. Aim 1: Determine the efficacy of AMR-MeOAc in the presence or absence of low dose FL118 on pancreatic cancer cell growth, apoptosis, and modulation of proteins in the relevant signaling pathways. Aim 2: Determine the efficacy of AMR-MeOAc in the presence or absence of low dose FL118 using human pancreatic cancer cell line-derived xenograft models. Aim 3: Determine the efficacy of AMR-MeOAc in the presence or absence of low dose FL118 against xenografts directly derived from patient pancreatic cancer tissues. Pancreatic cancer with K-ras gain-of-function mutations and aberrant expression of one or more antiapoptotic proteins (survivin, Mcl-1, XIAP, cIAP2) is hard to treat. This project may develop a novel and targeted combination strategy to effectively control this challenging and difficult-to-treat cancer.

摘要 该R 03试点项目的目标是证明创新组合治疗的概念验证 设计用于靶向具有K-ras突变和一种或多种 抗凋亡蛋白。K-ras癌基因突变和主要抗凋亡蛋白的异常表达（例如， 存活素、Mcl-1、XIAP、cIAP 2）在胰腺癌中对胰腺癌发展有很大贡献， 侵袭性（治疗抗性、转移和复发）。观察到K-ras功能获得性突变 >90%的胰腺癌患者。突变K-ras基因沉默诱导细胞凋亡并抑制 胰腺癌细胞生长、侵袭性、恶性肿瘤形成和异种移植肿瘤生长。然而，在这方面， 对于胰腺癌K-ras突变没有有效的靶向治疗。使用一种新 K-ras突变细胞与正常细胞的筛选方法， 药用植物Amoora rohituka进行了鉴定，并使用半合成方法产生了50多个衍生物 从15支安打中接近。然后使用K-ras突变细胞与正常细胞对这些化合物进行再筛选。 细胞AMR-Me和AMR-MeOAc被鉴定为选择性地针对K-ras的最有效的化合物 突变细胞我们以前的研究表明AMR-Me靶向K-ras通路，并且每天3 mg/kg， 28-对小鼠的24天治疗没有显示出明显的毒性，而它延长了白血病小鼠的存活。我们计划 将联合收割机AMR-MeOAc（最佳选择性）与我们的新型化合物FL 118结合，FL 118选择性抑制存活素， Mcl-1、XIAP和cIAP 2，用于测试这种新型组合靶向治疗方案。其原先已经 已经显示K-ras沉默与吉西他滨的组合显著减少小鼠中的肿瘤体积 与单独的单一药剂相比。因此，我们假设抑制两种突变的K-ras基因， 而主要的抗凋亡蛋白与新型药物AMR-MeOAc和FL 118将导致一种新的抗凋亡蛋白。 显著增强诱导胰腺癌细胞凋亡和抑制肿瘤生长 比单独使用任何一种药物的抑制效果都要好。该项目提出了以下三个具体目标。 目的1：确定AMR-MeOAc在存在或不存在低剂量FL 118的情况下对胰腺癌的功效。 癌细胞生长、凋亡和相关信号通路中蛋白质的调节。 目的2：使用人类细胞在低剂量FL 118存在或不存在的情况下确定AMR-MeOAc的功效。 胰腺癌细胞系衍生的异种移植模型。 目的3：确定在存在或不存在低剂量FL 118的情况下AMR-MeOAc针对以下的功效： 直接来源于患者胰腺癌组织的异种移植物。 胰腺癌伴K-ras功能获得性突变和一种或多种抗凋亡基因异常表达 蛋白（生存素，Mcl-1，XIAP，cIAP 2）是很难治疗的。该项目可能会开发一种新的，有针对性的 因此，我们需要采取联合策略来有效控制这种具有挑战性和难以治疗的癌症。