Targeting Glycolytic Control of Melanoma Cell Survival

描述（由申请方提供）：改变的存活信号传导是恶性黑色素瘤细胞的标志，导致对药物诱导的细胞凋亡的抗性。先前的研究表明，癌细胞的存活取决于甲基乙二醛（MG）对细胞死亡途径的调节，甲基乙二醛是糖酵解的反应性副产物，通过翻译后修饰和参与细胞存活的靶蛋白的功能改变起作用。我们的初步数据表明，热休克蛋白27（Hsp 27）是唯一的目标MG-加合在人黑色素瘤和MG-拮抗剂选择性诱导人黑色素瘤细胞凋亡和化疗增敏。在这个建议中，我们将测试的假设，糖酵解控制黑色素瘤细胞的生存是由MG加合热休克蛋白27（MG-Hsp 27），一种新的治疗目标，适合小分子调制特定的MG拮抗剂介导。首先，将通过绘制人黑色素瘤MG-Hsp 27蛋白质组图在分子、细胞和组织水平上表征MG-Hsp 27（具体目标#1）。将在黑素瘤细胞系和组织中确定通过磷酸化和MG-加合的人黑素瘤Hsp 27翻译后修饰的确切结构和位点。MG-Hsp 27随后将被验证为用于调节黑素瘤细胞存活的分子靶标（具体目标#2）。将阐明MG-Hsp 27的生存信号传导的分子机制，并通过MG-Hsp 27的代谢、药理学和遗传调节实现靶点验证。接下来，基于细胞筛选小分子MG拮抗剂的靶标调节效力和抗黑素瘤活性将鉴定一系列领先的生物活性MG-Hsp 27抑制剂（具体目标#3）。最后，将在异种移植小鼠黑色素瘤模型中使用MG拮抗剂作为单一药剂或与其他化疗剂组合测试MG拮抗剂的化疗潜力的原理证明证据（具体目标#4）。成功完成拟议的研究将确定一个独特的代谢脆弱性的癌细胞，可以攻击的小分子拮抗剂。铺设说明：黑色素瘤是一种高度侵袭性的肿瘤，起源于人类皮肤中的色素产生细胞，其发病率不断增加，目前超过任何其他癌症。我最近的研究表明，糖能量代谢的副产品甲基乙二醛可能是人类黑色素瘤细胞的致命弱点。该研究旨在阐明甲基乙二醛调节黑色素瘤细胞存活的分子机制，并测试甲基乙二醛的拮抗剂作为新型抗黑色素瘤治疗剂。

DESCRIPTION (provided by applicant): Altered survival signaling is a hallmark of malignant melanoma cells resulting in resistance to drug-induced apoptosis. Previous studies suggest that cancer cell survival depends on the modulation of cell death pathways by methylglyoxal (MG), a reactive by-product of glycolysis that acts by posttranslational modification and functional alteration of target proteins involved in cellular survival. Our preliminary data indicate that heat shock protein 27 (Hsp27) is the exclusive target of MG-adduction in human melanoma and that MG-antagonists selectively induce apoptosis and chemosensitization in human melanoma cells. In this proposal we will test the hypothesis that the glycolytic control of melanoma cell survival is mediated by MG adducted heat shock protein 27 (MG-Hsp27), a novel therapeutic target amenable to small molecule modulation by specific MG-antagonists. First, MG-Hsp27 will be characterized at the molecular, cellular, and tissue level by mapping the human melanoma MG-Hsp27 proteome (specific aim #1). The exact structure and site(s) of human melanoma Hsp27 posttranslational modification by phosphorylation and MG-adduction will be determined in melanoma cell lines and tissue. MG-Hsp27 will then be validated as a molecular target for the modulation of melanoma cell survival (specific aim #2). The molecular mechanism of survival signaling by MG-Hsp27 will be elucidated, and target validation will be achieved by metabolic, pharmacological, and genetic modulation of MG-Hsp27. Next, cell-based screening of small molecule MG antagonists for potency of target modulation and anti-melanoma activity will identify a lead series of bioactive MG-Hsp27-inhibitors (specific aim #3). Finally, proof-of-principle evidence for chemotherapeutic potential of MG-antagonists will be tested in a xenograft mouse melanoma model using MG-antagonists as single agents or in combination with other chemotherapeutic agents (specific aim #4). Successful completion of the proposed research will identify a unique metabolic vulnerability of cancer cells that can be attacked by small molecule antagonists. Lay description: Melanoma, a highly aggressive tumor that originates from pigment producing cells in human skin, has an increasing incidence that currently surpasses that of any other cancer. My recent research suggests that a byproduct of sugar energy metabolism called methylglyoxal may represent an Achilles heel of human melanoma cells. The proposed research aims to elucidate the molecular mechanism that regulates melanoma cell survival by methylglyoxal and to test antagonists of methylglyoxal as novel anti-melanoma therapeutics.