TAS::75 0849::TAS MULTIPLEXED ASSAYS FOR AKT/M TOR PATHWAY - PHASE I SBIR TOPIC

PTEN/Akt/mTORC1&2信号通路是细胞存活、生长和增殖、血管生成、细胞代谢、细胞迁移和侵袭以及葡萄糖摄取的重要调节因子。一些临床试验中的抗癌药物正在针对这一途径。在第一阶段，我们将建立可靠、准确、定量和灵敏的多重ELISA/qPCR检测方法(ELISA和定量PCR相结合)，以准确监测抗癌药物雷帕霉素和LY294002暴露的细胞中途径蛋白的蛋白水平和翻译后修饰。将生产酶联免疫吸附试验/定量聚合酶链式反应试剂盒，并将制定书面的标准操作规程。在第二阶段，我们将进一步改进检测方法，并使用它来量化多种癌症临床样本中含有重要的翻译后修饰组合的靶分子的数量。一种灵敏、定量和标准化的多重酶联免疫吸附试验/定量聚合酶链式反应的出现将有助于临床医生和研究人员对多种疾病中的PTEN/Akt通路蛋白成员进行分析，并预测癌症治疗的疗效。

PTEN/Akt/mTORC 1&2 signal pathway is a critical regulator of cell survival, cell growth and proliferation, angiogenesis, cellular metabolism, cell migration and invasion and glucose uptake. A number of anticancer drugs in clinical trials are targeting this pathway. In Phase I, we will develop reliable, accurate, quantitative and sensitive multiplexed hybrid ELISA/qPCR assay (ELISA combined with quantitative PCR) to accurately monitor protein level and post-translational modifications of the pathway proteins in cells exposed to anticancer drugs rapamycin and LY294002. The ELISA/qPCR kit will be manufactured and written SOPs will be created. In Phase II, we will further improve the assay and use it to quantify the number of target molecules containing important combinations of post-translational modifications in clinical samples of a wide range of cancers. The availability of a sensitive, quantitative and standardized multiplexed ELISA/qPCR will aid clinicians and researchers to profile PTEN/Akt pathway protein members in a number of diseases and predict the efficacy of cancer treatment.