Genomic screening to identify novel stress-inducing chemotherapies for carcinoma

尽管口腔鳞状细胞癌患者的手术和放射治疗技术不断进步， 口腔鳞状细胞癌（OSCC）的生存率在过去的二十年中一直没有提高，这表明我们 治疗病人的能力已经达到了一个平台。我们已经报道了蛋白酶体抑制诱导ER 在HNSCC细胞系中应激CHOP介导的凋亡，包括几种顺铂耐药细胞。儿 应激导致称为未折叠蛋白质反应（UPR）的生理反应，其包括 不同的平行遗传程序，试图恢复稳态蛋白质折叠或导致细胞凋亡， 压力是长期的或强大的。我们假设我们可以使用高通量筛选（HTS）， 不同的化学库，以确定新的小分子CHOP激活剂，以诱导OSCC细胞凋亡 细胞我们已经开发了一种基于互补细胞的检测方法，该方法使用稳定转染的CMQ-K1细胞， 分别报告了PERK/eIF 2D/CH 0 P（凋亡）和IRE 1/XBP 1（适应性）通路。 普遍定期审议。鉴定仅特异性激活PERK/eIF 2n/CHOP而不激活1 RE 1/XBP 1的化合物， 使我们能够排除许多通常会干扰蛋白质折叠的化合物。已经有65个被击中的化合物 确定了激活CHOP但不激活XBP 1报告基因的化学特征， 合成类比和治疗开发。我们建议测试这65种化合物的能力， 沿着与市售类似物系列在一组小鼠中诱导CHOP表达和凋亡。 口腔鳞癌细胞。最有希望的候选者将在体内进行评估，在一个良好的特点rbd^ht 异种移植模型拟议研究的长期目标是阐明这些新的 诱导CHOP和抑制生长或诱导OSCC细胞凋亡的小分子，并进一步筛选 非常大的不同化学探针库与我们的互补细胞为基础的筛选，小分子 通过这些研究确定的UPR激动剂将提供一种方便的成本有效的治疗方法， 患有治疗仅限于最低限度有效化疗的疾病的患者， 有害的辐射或痛苦的手术，往往会干扰他们说话，咀嚼甚至呼吸的能力。

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.