Exploiting synthetic-lethal interactions to target triple-negative breast cancers

项目总结/摘要 目前没有针对三阴性（TN）乳腺癌的靶向治疗策略， 最难治疗的一种乳腺癌，它不会过度表达人类表皮生长因子 受体2（HER 2），并且缺乏雌激素和孕激素受体的表达。因此， 迫切需要加深我们对这种侵袭性乳腺癌亚型的理解， 治疗干预的临床相关目标。我们最近发现， 转录因子MYC以及MYC依赖性信号通路在 原发性人类三阴性乳腺肿瘤。此外，我们发现MYC激活与 患者预后不佳，这表明MYC通路可能在驱动糖尿病的发生中发挥重要作用。 这些恶性肿瘤的形成。 我们如何杀死MYC驱动的TN肿瘤？由于MYC是一种转录因子， 可直接抑制其活性的小分子抑制剂不能用于临床应用。一个替代 选择性杀死MYC驱动的肿瘤的方法是利用“合成-致死”相互作用的存在。 我们的研究小组以前采用了细胞周期偏向的方法，发现抑制有丝分裂激酶， 细胞周期蛋白依赖性激酶（CDK）1导致细胞凋亡工程过度表达MYC。的 这种细胞死亡的机制涉及促凋亡BCL-2家族成员BIM的上调。我们 随后使用该方法处理具有升高的MYC表达的TN细胞系和异种移植肿瘤。 这些观察结果表明，MYC依赖的合成-致死相互作用存在，重要的是， 靶向选择性杀死MYC驱动的肿瘤。 本研究的目的1是进一步研究小分子CDK的临床应用潜力 对MYC驱动的TN癌症的抑制，特别是与抗凋亡的临床抑制剂组合 BCL-2成员。这是基于我们的新假设，因为CDK抑制上调BIM， 激活线粒体内源性途径，CDK和抗凋亡抑制剂的联合使用 BCL-2家族成员可显著提高细胞死亡率。目标2将描述一个新的 发现MYC激活和Pim 1抑制之间的合成致死相互作用，Pim 1是一种非必需的， 先前显示与MYC基因相互作用的激酶。目标3将进行高通量小 分子筛选以鉴定能够诱导MYC依赖性合成致死性的潜在先导分子 在乳腺细胞中。我们将使用模型人类乳腺癌的组合进行这些实验， 上皮细胞、基因确定的癌细胞系和一组新型人鼠原位肿瘤 嫁接模型如果成功，拟议的研究不仅将扩大我们对MYC生物学的知识， 还将提供新的治疗概念，以再次测试TN肿瘤患者。

PROJECT SUMMARY/ABSTRACT No targeted therapeutic strategies are currently available against triple-negative (TN) breast cancer, the most difficult-to-treat form of breast cancer, which does not overexpress human epidermal growth factor receptor 2 (HER2) and lacks the expression of the estrogen and progesterone receptors. Thus, there is an urgent need in deepening our understanding of this aggressive breast cancer subtype and identifying clinically relevant targets for therapeutic intervention. We have recently discovered that an oncogenic transcription factor MYC as well as the MYC-dependent signaling pathways are significantly up-regulated in primary human triple-negative breast tumors. In addition, we found that MYC activation is associated with patientsʼ poor prognosis suggesting that the MYC pathways may play a fundamental role in driving the formation of these aggressive tumors. How can we kill MYC-driven TN tumors? Because MYC is a transcription factor, rationally designed small molecule inhibitors that can directly inhibit its activity are not available for clinical use. An alternative approach in selectively killing MYC-driven tumors is to exploit the existence of “synthetic-lethal” interactions. Our group previously took a cell cycle-biased approach and discovered that inhibition of the mitotic kinase cyclin-dependent kinase (CDK) 1 resulted in apoptosis in cells engineered to overexpress MYC. The mechanism of such cell death involved an up-regulation of a pro-apoptotic BCL-2 family member BIM. We subsequently used this approach to treat TN cell lines and xenograft tumors with elevated MYC expression. These observations suggest that MYC-dependent synthetic-lethal interactions exist, and importantly, can be targeted to selectively kill MYC-driven tumors. Aim 1 of this proposed research will further study the clinical potential of small molecule CDK inhibition against MYC-driven TN cancers particularly in combination with clinical inhibitors of anti-apoptotic BCL-2 members. This is based on our novel hypothesis that, because CDK inhibition up-regulates BIM and activates mitochondrial intrinsic pathway, the combined use of the inhibitors for CDK and anti-apoptotic BCL-2 family members may significantly enhance the rate of cell death. Aim 2 will characterize a newly discovered synthetic-lethal interaction between MYC activation and inhibition of Pim1, a non-essential, kinase previously shown to genetically interact with MYC. Aim 3 will conduct a high-throughout small molecule screen to identify potential lead molecules capable of inducing MYC-dependent synthetic lethality in mammary cells. We will carry out these experiments using a combination of model human mammary epithelial cells, genetically defined cancer cell lines, and a panel of novel human-in-mouse orthotopic tumor grafts models. If successful, the proposed research will not only expand our knowledge on MYC biology but will also provide novel therapeutic concepts to be tested again patients with TN tumors.