Development of Small Molecule Radiation Sensitizers

小分子放射增敏剂的开发

基本信息

批准号：
8518497
负责人：
MICHAEL L. FREEMAN
金额：
$ 1.56万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-12-11 至 2014-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8518497
关键词：
A549 Acute Apoptosis Biochemical Biological Assay Blood Vessels Bolus Infusion CD34 gene Cell Proliferation Cell membrane Colon Carcinoma Coupled DNA biosynthesis Data Development Disease Disulfides Dorsal Endothelial Cells Enzymes Epithelial Cells Exhibits Genetic Genetic Screening Goals Human Injection of therapeutic agent Ionizing radiation Lead Lewis Lung Carcinoma Malignant Neoplasms Mediating Modeling Molecular Molecular Target Morphogenesis Mus Neoplasm Metastasis Non-Small-Cell Lung Carcinoma Oxidation-Reduction Proteins Radiation Radiation Tolerance Radiation therapy Radiation-Sensitizing Agents Research Risk Screening Result Skin Structure Sulfhydryl Compounds Testing Therapeutic Toxic effect Tumor-Associated Vasculature Xenograft Model analog angiogenesis cancer cell cell motility chemical genetics clinical efficacy density design inhibitor/antagonist irradiation matrigel migration neoplastic cell novel public health relevance scaffold small hairpin RNA small molecule small molecule libraries tumor tumor growth

项目摘要

DESCRIPTION (provided by applicant): We synthesized a novel synthetic chemical library and used it in a forward chemical genetics screen designed to identify small molecule inhibitors of endothelial morphogenesis. This screen resulted in the identification of structurally-related small molecules that inhibited endothelial cell proliferation, migration, the ability to form tubule-like structures in matrigel, as well as neo- angiogenesis visualized using a dorsal skin fold vascular window chamber. Biochemical analysis revealed that the novel small molecules inhibited ENOX1, a plasma membrane- associated enzyme that exhibits protein disulfide-thiol interchange activity. While ENOX activity is important for cellular proliferation, the molecular mechanisms are not well understood. shRNA-mediated inhibition of ENOX activity in HUVECs inhibited endothelial cell migration and formation of tubule-like structures in matrigel, reproducing the effects of the small molecule inhibitors. Small molecule inhibition of ENOX1 was associated with a significant increase in endothelial cell apoptosis induced by ionizing radiation. Colony formation assays demonstrated that these small molecules increased the radiation sensitivity of endothelial cells. In contrast, the radiation sensitivity of tumor epithelial cells was unaffected by the small molecules. Administration of a small molecule ENOX1 inhibitor prior to fractionated X-irradiation produced a statistically significant decrease in the number and density of CD34 expressing Lewis Lung Carcinoma (LLC) tumor-associated microvasculature. Use of LLC and human HT29 colon carcinoma tumor models demonstrated that the small molecule ENOX1 inhibitors coupled with fractionated X-irradiation produced a statistically significant increase in tumor growth delay compared to irradiation alone. No evidence of acute toxicity was observed over a 30 day interval when mice received a bolus injection of 120 mg/kg. These preliminary data support the hypothesis that ENOX1 represents a rationale molecular target for increasing the ability of ionizing radiation to control tumor growth. The short term goal of this application is to valid this hypothesis. The long term goal is to develop these small molecule inhibitors into a radiation sensitizer that exhibits clinical efficacy. PUBLIC HEALTH RELEVANCE: A forward chemical genetics screen identifies small molecule inhibitors of endothelial morphogenesis. Biochemical analysis revealed that these novel small molecules inhibited the enzyme ENOX1, increased the radiation sensitivity of endothelial cells, increased radiation- mediated suppression of tumor-associated vasculature, thereby suppressing tumor growth. These preliminary data support the hypothesis that ENOX1 represents a rationale molecular target for increasing the ability of ionizing radiation to control tumor growth.

描述（由申请人提供）：我们合成了一个新型的合成化学文库，并将其用于旨在鉴定内皮形态发生的小分子抑制剂的正向化学遗传学筛选中。该屏幕导致鉴定与结构相关的小分子，这些小分子抑制了内皮细胞增殖，迁移，在Matrigel中形成小管状结构的能力以及使用背面皮肤折叠式血管窗口室可视化的新血管生成。生化分析表明，新型的小分子抑制了ENOX1，ENOX1是一种质膜相关的酶，表现出蛋白质二硫键 - 硫醇互换活性。虽然ENOX活性对于细胞增殖很重要，但分子机制尚不清楚。 SHRNA介导的HUVEC中eNOX活性的抑制作用抑制了内皮细胞迁移和Matrigel中小管状结构的形成，从而再现了小分子抑制剂的作用。 ENOX1的小分子抑制与电离辐射引起的内皮细胞凋亡的显着增加有关。菌落形成分析表明，这些小分子增加了内皮细胞的辐射灵敏度。相反，肿瘤上皮细胞的辐射敏感性不受小分子的影响。在分离X辐射之前，给予小分子ENOX1抑制剂在表达Lewis Lung Carcinoma（LLC）肿瘤相关的微腔的CD34的数量和密度上统计学上显着降低。使用LLC和人类HT29结肠癌肿瘤模型的使用表明，与单独的辐照相比，与分馏的小分子ENOX1抑制剂与分馏X-辐射相结合的肿瘤生长延迟在统计上显着增加。在小鼠注射120 mg/kg时，在30天的间隔内未观察到急性毒性的证据。这些初步数据支持以下假设：ENOX1代表了提高电离辐射控制肿瘤生长的能力的基本原理分子靶标。该应用程序的短期目标是有效此假设。长期目标是将这些小分子抑制剂开发为具有临床功效的辐射敏化剂。公共卫生相关性：正向化学遗传学筛选鉴定出内皮形态发生的小分子抑制剂。生化分析表明，这些新型的小分子抑制了酶ENOX1，增加了内皮细胞的辐射敏感性，增加了辐射介导的抑制与肿瘤相关的脉管系统的抑制，从而抑制了肿瘤的生长。这些初步数据支持以下假设：ENOX1代表了提高电离辐射控制肿瘤生长的能力的基本原理分子靶标。