High Throughput Chemical Genomic screening to identify novel ER stress-inducing

高通量化学基因组筛选以识别新型 ER 应激诱导物

• 批准号: 7642746
• 负责人: Andrew Michael Fribley
• 金额: $ 13.73万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-05 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7642746
• 关键词: Academia; Address; Adjuvant Therapy; Affect; Apoptosis; Apoptotic; Biological Assay; Breathing; Caring; Cell Death; Cell Line; Cells; Chemicals; Cisplatin; Development; Disease; Dose; Drug Interactions; Drug Kinetics; Enzymes; Funding; Genetic Programming; Genomics; Glean; Glycoproteins; Growth; Head and Neck Neoplasms; Head and Neck Squamous Cell Carcinoma; Head and neck structure; Human; In Vitro; Individual; Investigation; Laboratories; Lead; Libraries; Luciferases; Malignant Epithelial Cell; Mediating; Metabolic; Metabolism; Michigan; Molecular Target; Neoplasm Metastasis; Operative Surgical Procedures; Oxidation-Reduction; Pain; Pathway interactions; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Physiological; Principal Investigator; Probability; Property; Proteasome Inhibition; Protein Biosynthesis; Proteins; Pump; Radiation; Radiation therapy; Reporter; Reporting; Resistance; Rodent; Screening procedure; Sensitivity and Specificity; Series; Signal Transduction; Stress; Structure; Survival Rate; Technology; Testing; Tumor Burden; Universities; Upper arm; Work; Xenograft Model; Xenograft procedure; analog; base; cellular targeting; chemotherapy; cost; cytotoxic; drug development; drug metabolism; high throughput screening; improved; in vivo; insight; killings; mouth squamous cell carcinoma; neoplastic cell; new therapeutic target; novel; protein expression; protein folding; public health relevance; research study; response; retinal rods; small molecule; small molecule libraries; success; therapeutic development; transcription factor CHOP; treatment strategy; tumor

DESCRIPTION (provided by applicant): Despite technological advances in surgery and radiotherapy for patients who suffer from oral squamous cell carcinoma (OSCC), the survival rate has remained un-improved during the last two decades indicating our ability to treat patients has reached a plateau. We have reported that proteasome inhibition induced ER stress and C/EBP-homologous protein- (CHOP) mediated apoptosis in a variety of HNSCC cell lines, including several that are not sensitive to cisplatin. ER stress leads to a physiological response known as the unfolded protein response (UPR) which consists of distinct parallel genetic programs that attempt to restore homeostatic protein folding or lead to apoptosis if the stress is prolonged or robust. We hypothesized that we could use high throughput screening (HTS) with large diverse chemical libraries to identify novel small molecule activators CHOP to induce apoptosis in OSCC cells. We have developed a complementary cellbased assay using stably transfected CHO-K1 cells that individually report on the PERK/elF2o/CHOP (apoptotic) and the IRE1/XBP1 (adaptive) pathways of the UPR. Identifying compounds that specifically activate only PERK/elF2a/CHOP and not IRE1/XBP1 has allowed us to rule out many compounds that are generally toxic as well as those that alter general properties of protein synthesis and/or folding, such as Ca2+ or redox status. 65 "hit" compounds have been identified that robustly activated the CHOP but not the XBP1 reporter with chemical signatures amenable for synthetic analoging and pharmaceutical development. We propose to test the ability of these 65 compounds along with commercially available analog series to induce CHOP expression and apoptosis in a panel of OSCC cells. The most promising candidates will be evaluated in vivo in a well-characterized rodent xenograft model. The long-term objectives of the proposed studies are to elucidate the ability of these novel small molecules to induce CHOP and inhibit growth or induce apoptosis in OSCC cells and to further screen very large diverse libraries of chemical probes with our complementary cell-based screen. Small molecule UPR agonisits identified through these studies will provide a convenient cost-effective means of treating patients suffering from a disease for which therapies are limited to minimally effective chemotherapies, damaging radiation or painful surgeries that often interfere with their ability to speak, chew or even breathe. PUBLIC HEALTH RELEVANCE: The survival rate for patients with tumors in the head and neck region has not improved in over 20 years. We screened a chemical library of 66,000 small molecules to identify novel drug-like compounds. The proposed studies will determine the ability of "hits" from this screen to interfere with the growth of or kill head and neck tumor cells.

描述（由申请人提供）：尽管对患有口腔鳞状细胞癌（OSCC）的患者进行了手术和放疗的技术进步，但在过去的二十年中，生存率仍未得到改善，表明我们治疗患者的能力已经达到了高原。我们已经报道了蛋白酶体抑制诱导的ER应激和C/EBP同源蛋白（CHOP）在多种HNSCC细胞系中介导的凋亡，其中包括几种对顺铂不敏感的。 ER应激会导致一种称为展开的蛋白质反应（UPR）的生理反应，该反应由不同的平行遗传程序组成，该程序试图恢复稳态蛋白质折叠或导致稳定性（如果应力延长或稳定）导致凋亡。我们假设我们可以使用具有大型化学文库的高吞吐量筛选（HTS）来鉴定新型的小分子激活剂切碎以诱导OSCC细胞中的细胞凋亡。我们使用稳定转染的CHO-K1细胞开发了一种互补的细胞测定，该测定单独报告UPR的PERK/ELF2O/CHOP（凋亡）和IRE1/XBP1（自适应）途径。识别专门激活PERK/ELF2A/CHOP而不是IRE1/XBP1的化合物使我们能够排除许多通常有毒的化合物，以及那些改变蛋白质合成和/或折叠的一般特性的化合物，例如Ca2+或dedox状态。已经确定了65个“命中”化合物，可以牢固地激活CHOP，而不是具有可用于合成类似物和药物开发的化学特征的XBP1报告基因。我们建议测试这65种化合物的能力以及市售的模拟系列，以诱导OSCC细胞小组中的CHOP表达和凋亡。最有希望的候选者将在特征良好的啮齿动物异种移植模型中在体内评估。拟议研究的长期目标是阐明这些新型小分子诱导斩波和抑制生长或诱导OSCC细胞中的凋亡的能力，并通过基于互补的细胞筛选进一步筛选出非常大的化学探针文库。通过这些研究鉴定出的小分子UPR激动剂将提供一种方便的成本效益手段，用于治疗患有疾病的患者，该疾病仅限于最小有效的化学疗法，损害辐射或痛苦的手术，通常会干扰其说话，咀嚼甚至呼吸的能力。公共卫生相关性：在20多年来，头颈部肿瘤患者的生存率尚未提高。我们筛选了一个66,000个小分子的化学文库，以鉴定出新型药物样化合物。拟议的研究将确定从此屏幕中“命中”的能力，以干扰头颈肿瘤细胞的生长或杀死头颈肿瘤细胞的生长。