High Throughput Screening of Agents in Bladder Cancer Cell Lines

膀胱癌细胞系中药物的高通量筛选

基本信息

批准号：
9344189
负责人：
Piyush Agarwal
金额：
$ 8.72万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9344189
关键词：
Accounting Animal Model Binding Biological Assay Bladder Calmette-Guerin Bacillus Cancer cell line Caspase Cell Line Cell Proliferation Cells Cessation of life Data Diagnosis Disease Dose Drug usage Epithelial Explosion Formulation Funding Health Heat-Shock Proteins 90 Histone Deacetylase Inhibitor Immunohistochemistry In Vitro Intravesical Administration Life Malignant Neoplasms Malignant neoplasm of prostate Malignant neoplasm of urinary bladder Mesenchymal Mitomycins Muscle Operative Surgical Procedures Patients Pharmaceutical Preparations Pharmacotherapy Prevalence Publications Publishing Recurrent disease Renal carcinoma Robotics SN-38 Secure Topoisomerase-I Inhibitor Topotecan Treatment Efficacy United States Urologic Cancer Water Woman Work base burden of illness cancer therapy chemotherapy cytotoxicity flavopiridol gemcitabine high throughput screening in vivo inhibitor/antagonist intravesical material transfer agreement men mimetics novel novel therapeutics oncology pre-clinical prevent response targeted agent tumor

项目摘要

I initially chose two cell lines that represented the spectrum of bladder cancer cell lines: UMUC-3 and RT4. The former is a mesenchymal, high-grade, and a luminal tumor. The latter is an epithelial, low-grade, and a basal tumor. The cells were treated with 1,912 oncology-focused drugs using a 48 hr cell proliferation assay with an ATP-based readout (CellTiterGlo), and activity of the compounds in a dose response manner was calculated. Encouraged by the preliminary results showing HSP90 inhibitors, topoisomerase I inhibitors, and HDAC inhibitors all being effective, we extended the screen to 8 bladder cancer cell lines. We found similar results except now combinations were also explored. Some of the combinations that we hope to explore in animal models include a novel SMAC mimetic + gemcitabine or mitomycin C. We also identified 8 novel agents that may function as intravesical agents and are currently evaluating their use as well. Finally, we identified a topotecan derivative (SN-38) from the screen that looks promising. We are asking our collaborators to bind existing Pan-IR700 conjugate to SN-38 to increase the cytotoxicity of this new conjugate. During this past year, we have focused our efforts on two new novel compounds: flavopiridol and bardoxolone methyl. For flavopiridol, we have established in vitro and in vivo activity across a wide range of bladder cancer cell lines. In addition, we have secured a material transfer agreement with a company to obtain a new water soluble formulation of flavopiridol that we will evaluate for possible intravesical administration. The pre-clinical work is pending immunohistochemistry analysis prior to submission for publication. For bardoxolone methyl, we have established that the drug is active in vitro especially in spheroid cultures. Furthermore, we have worked out a potential mechanism of action. Ongoing in vivo work demonstrates activity across different bladder cancer cell lines. This work will be published in conjunction with a summary of the high throughput screening data.

我最初选择了两种代表膀胱癌细胞系的细胞系：UMUC-3和RT4。前者是间充质，高级和腔肿瘤。后者是上皮，低度和基底肿瘤。使用基于ATP的读数（CellTiterGlo）的48小时细胞增殖测定法（CellTiterGlo），用1,912个以肿瘤学为中心的药物处理细胞，并计算化合物的活性。受到初步结果的鼓励，显示HSP90抑制剂，拓扑异构酶I抑制剂和HDAC抑制剂均有效，我们将筛网扩展到了8个膀胱癌细胞系。我们发现了类似的结果，除了现在还探索了组合。我们希望在动物模型中探索的某些组合包括一种新型的SMAC模拟 +吉西他蛋白滨或丝裂霉素C。我们还确定了8种可能充当静脉内药物的新型药物，目前也正在评估它们的使用。最后，我们从屏幕上鉴定了一个看起来很有希望的拓扑衍生物（SN-38）。我们要求我们的合作者将现有的Pan-IR700共轭物绑定到SN-38，以增加这种新结合物的细胞毒性。在过去的一年中，我们将精力集中在两种新的新颖化合物上：黄叶替葡萄托和bardoxolone甲基。对于黄叶肽，我们已经在各种膀胱癌细胞系中建立了体外和体内活性。此外，我们已经与公司获得了一项重大转移协议，以获得黄虫寡醇的新水溶性公式，我们将评估可能的静脉内给药。临床前的工作是在提交出版之前等待免疫组织化学分析。对于Bardoxolone甲基，我们已经确定该药物在体外是活性的，尤其是在球体培养物中。此外，我们已经确定了一种潜在的作用机制。正在进行的体内工作表明了不同膀胱癌细胞系的活性。这项工作将与高通量筛选数据的摘要一起发布。