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

项目概要/摘要 控制肿瘤复发和扩散是乳腺癌的主要挑战。这种情况进一步恶化 许多流行病学研究表明，患有2型糖尿病的妇女在 患乳腺癌、复发和死于乳腺癌的风险显著高于 他们不是糖尿病患者。研究表明，与2型糖尿病相关的高胰岛素血症是一个重要的贡献， 在乳腺癌患者中观察到的死亡率的因素，这表明对有效治疗的重要需求， 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cdk 4敲- 我们的小鼠没有发生由ERBB 2或RAS癌基因驱动的乳腺癌，这表明CDK 4/6 抑制剂可能是某些乳腺癌的有效治疗剂。批准CDK 4/6抑制剂作为 乳腺癌治疗学证实了这一理论。另一种在胰岛素信号传导中起关键作用的激酶 而肿瘤的进展是NUAK 1/ARK 5，属于AMPK基因家族，调节代谢。 NUAK 1/ARK 5在整个生物体中发挥的正常生理作用是使用 肌肉特异性基因敲除小鼠显示，NUAK 1通过以下途径控制葡萄糖代谢： 胰岛素信号的调节。因此，当这些基因敲除小鼠被喂食高脂肪饮食时， 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， 与对照小鼠相比，较高浓度的肌糖原表明， ARK 5/NUAK 1可以克服高血糖的影响。有趣的是，ARK 5最初被认为是 作为一种转移基因，其过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 CDK 4和ARK 5的抑制剂（ON 123300），这是一种非常有效的乳腺肿瘤生长抑制剂。在 此外，该化合物对高糖饮食诱导的肿瘤发展显示出深刻的影响， 转移的果蝇模型系统。在本申请中，我们建议将这些研究扩展到小鼠模型 高胰岛素血症（MKR小鼠），由LeRoith博士和PDX三阴性乳腺癌模型开发 （TNBC）由Irie博士开发。目的：（1）检测ON 123300对人肝癌细胞生长和转移的影响 表现出高胰岛素血症、糖尿病前期表型的MKR小鼠中乳腺肿瘤的发生率;（2）检查 高胰岛素血症对PDX乳腺肿瘤生长和转移的影响， ARK 5的最高相对水平以检查ON 123300的治疗价值;以及（3）使用新的 开发了多重激酶抑制剂珠（MIB）和“癌症工具包功能获得”（CTK）技术 以确定乳腺癌细胞在长期暴露后是否对ON 123300产生抗性， 确定信号通路的性质，这可能是这种耐药性的根本原因。

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.