Targeting cell cycle and metabolic pathways of high risk breast cancers using mouse models of hyperinsulinemia

项目摘要/摘要 控制肿瘤的复发和扩散是乳腺癌的一大挑战。这种情况会进一步恶化 对于患有糖尿病的妇女来说，因为许多流行病学研究表明，患有2型糖尿病的妇女 与女性相比，患乳腺癌、复发和死于乳腺癌的风险明显更高 不是糖尿病患者。研究表明，与2型糖尿病相关的高胰岛素血症是一个重要因素 乳腺癌患者死亡率的因素，表明需要有效的治疗方法 在高胰岛素血症条件下抑制肿瘤细胞增殖。我们早期的研究表明，CDK4会敲打- Out小鼠未发生ERBB2或RAS癌基因驱动的乳腺癌，提示CDK4/6 对于某些乳腺癌，抑制剂可能是有效的治疗剂。CDK4/6抑制剂被批准为 乳腺癌治疗学证实了这一理论。另一种在胰岛素信号转导中起关键作用的激酶 肿瘤进展为NUAK1/ARK5，属于调节代谢的AMPK基因家族。 研究了NUAK1/ARK5在整个生物体中的正常生理作用 肌肉特异性基因敲除小鼠，表明NUAK1通过 胰岛素信号的调节。因此，当这些基因敲除的小鼠被喂以高脂肪饮食时，它们 表现出较低的空腹血糖水平，较高的葡萄糖耐量，较高的胰岛素敏感性，以及 与对照组小鼠相比，肌糖原浓度更高，这表明抑制 ARK5/NUAK1可以克服高血糖的影响。有趣的是，ARK5最初被发现 作为一种转移基因，其过度表达可促进多种肿瘤的转移 类型。为了达到抑制乳腺肿瘤细胞生长和代谢的目的，我们开发了一种有效的双重 CDK4和ARK5的抑制剂(ON123300)，它是一种非常有效的乳腺肿瘤生长抑制物。在……里面 此外，这种化合物对高糖饮食诱导的肿瘤发生和发展具有深远的影响。 果蝇模型系统中的转移。在本申请中，我们建议将这些研究扩展到小鼠模型 LeRoith博士建立的高胰岛素血症(MKR小鼠)和PDX三阴性乳腺癌模型 (TNBCS)由Irie博士开发。目的：(1)检测ON123300对肿瘤生长和转移的影响 对表现为高胰岛素血症、糖尿病前期表型的MKR小鼠的乳腺肿瘤进行研究；(2)检查 高胰岛素血症对PDX乳腺肿瘤生长和转移的影响及其模型的应用 检测ON123300治疗价值的最高相对水平ARK5；以及(3)使用新的 已开发的多重激酶抑制物珠(MIB)和“癌症工具包功能增益”(CTK)技术 确定乳腺癌细胞在长期暴露后是否对ON123300产生耐药性 确定可能是这种抗性的根本原因的信号通路的性质。

Project Summary/Abstract Managing tumor recurrence and spread is a major challenge in breast cancer. This scenario is further aggravated for women with diabetes as numerous epidemiological studies show that women with type 2 diabetes are at significantly greater risk of developing, relapsing with, and dying from breast cancer compared to women who are not diabetic. Studies show that hyperinsulinemia associated with type 2 diabetes is a significant contributing factor for the mortality seen in breast cancer patients, suggesting an important need for effective therapies that inhibit tumor cell proliferation under hyperinsulinemic conditions. Our early studies showed that cdk4 knock- out mice fail to develop breast cancers driven by the ERBB2 or RAS oncogenes suggesting that CDK4/6 inhibitors may be effective therapeutic agents for certain breast cancers. The approval of CDK4/6 inhibitors as breast cancer therapeutics validated this theory. Another kinase which plays a critical role in insulin signaling and cancer progression is NUAK1/ARK5 which belongs to the AMPK gene family which regulate metabolism. The normal physiological role played by NUAK1/ARK5 in the whole organism was studied using muscle-specific knock-out mice which showed that NUAK1 controls glucose metabolism through regulation of the insulin signaling. Thus, when these knock-out mice were fed a high fat diet, they exhibited a lower fasting blood glucose level, greater glucose tolerance, higher insulin sensitivity, and higher concentrations of muscle glycogen compared to control mice suggesting that inhibition of ARK5/NUAK1 can overcome the effects of hyperglycemia. Interestingly, ARK5 was originally identified as a metastasis gene and its over-expression has been shown to promote metastasis of several tumor types. To achieve the goal of inhibiting breast tumor cell growth and metabolism, we developed a potent dual inhibitor of CDK4 and ARK5 (ON123300), which was a very effective inhibitor of breast tumor growth. In addition, this compound has shown a profound effect on high sugar diet-induced tumor development and metastasis in a Drosophila model system. In this application, we propose to extend these studies to mouse models of hyperinsulinemia (MKR mice), developed by Dr. LeRoith and PDX models of Triple Negative Breast cancer (TNBCs) developed by Dr. Irie. The aims are: (1) To test the effects of ON123300 on the growth and metastasis of mammary tumors in MKR mice which exhibit hyperinsulinemic, pre-diabetic phenotype; (2) To examine the effects of hyperinsuminemia on the growth and metastasis of PDX breast tumors and utilize the models with the highest relative levels of ARK5 to examine the therapeutic value of ON123300; and (3) To use the newly developed Multiplexed-kinase Inhibitor Beads (MIB) and “Cancer Toolkit gain-of-function” (CTK) technologies to determine whether breast cancer cells develop resistance to ON123300 upon prolonged exposure and to determine the nature of signaling pathways that might be the root cause of such resistance.