Genomic screening to identify novel stress-inducing chemotherapies for carcinoma

尽管口腔鳞状细胞癌患者在手术和放射治疗方面取得了技术进步 癌症(OSCC)，生存率在过去的二十年里一直没有改善，这表明我们的 治疗病人的能力已经达到了平台期。我们已经报道了蛋白酶体抑制诱导内质网 应激CHOP介导的HNSCC细胞株的凋亡，包括几个顺铂耐药的细胞。呃 应激导致一种称为未折叠蛋白反应(UPR)的生理反应，它包括 独特的并行遗传程序，试图恢复体内平衡的蛋白质折叠或在以下情况下导致细胞凋亡 压力是持久的或强烈的。我们假设我们可以使用高通量筛选(HTS) 不同化学文库寻找诱导口腔鳞癌细胞凋亡的新型小分子CHOP激活剂 细胞。我们已经开发了一种基于细胞的互补分析方法，使用稳定转染的CMQ-K1细胞 PERK/elF2D/CH0P(凋亡)和IRE1/XBP1(适应性)通路的单独报告 UPR。鉴定只激活PERK/elF2n/CHOP而不激活1RE1/XBP1的化合物 使我们能够制造出许多通常扰乱蛋白质折叠的化合物。65种热门化合物已经被 已确定激活了印章，但不是具有符合以下条件的化学签名的XBP1报告器 合成类比和治疗发展。我们建议测试这65种化合物的能力 以及商业上可用的类似物系列来诱导CHOP表达和细胞凋亡 口腔鳞癌细胞。迄今最有希望的候选人将在具有良好特征的rbd？ht中进行活体评估。 异种移植模型。拟议研究的长期目标是阐明这些小说的能力 诱导口腔鳞癌细胞CHOP和抑制生长或诱导细胞凋亡的小分子及进一步筛选 非常大的不同的化学探针库，带有我们的基于细胞的补充屏幕，小分子 通过这些研究确定的UPR疼痛将提供一种方便、成本效益高的治疗方法。 患有一种疾病的患者，其治疗方法仅限于最低有效的化疗， 破坏性的辐射或痛苦的手术往往会干扰他们的说话、咀嚼甚至呼吸能力。

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 CHOP-mediated apoptosis in HNSCC cell lines, including several that are cisplatin-resistent. 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 diverse chemical libraries to identify novel small molecule CHOP activators to induce apoptosis in OSCC cells. We have developed a Complementary cell-based assay using stably transfected CMQ-K1 cells that individually report on the PERK/elF2D/CH0P (apoptotic) and the IRE1/XBP1 (adaptive) pathways of the UPR. Identifying compounds that specifically activate only PERK/elF2n/CHOP and not 1RE1/XBP1 has allowed us to aile out many compounds that generally perturb protein folding. 65 hit compounds have been identified that activated the CHOP but not the XBP1 reporter with chemical signatures amenable for synthetic analoging and therapeutic 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 hiost promising candidates will be evaluated in vivo in a well-characterized rbd^ht 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 pfovide a convenient cost-effective rneans 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.