Identification of potentially active tyrosine kinases in lung cancer

肺癌中潜在活性酪氨酸激酶的鉴定

• 批准号: 9153902
• 负责人: Udayan Guha
• 金额: $ 40.54万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9153902
• 关键词: Antibodies; Biological Assay; Clinical Trials; Collaborations; Data; EPHA2 gene; Epidermal Growth Factor Receptor; Genes; Goals; Human; Institutes; Intramural Research Program; KRAS2 gene; Lung; Lung Adenocarcinoma; Malignant Neoplasms; Malignant neoplasm of lung; Measures; Memorial Sloan-Kettering Cancer Center; Mutate; Mutation; NTRK1 gene; Patients; Phosphotransferases; Phosphotyrosine; Protein Tyrosine Kinase; Sequence Analysis; Somatic Mutation; Structure of parenchyma of lung; Transcript; Tumor Biology; Tumor Tissue; Tyrosine; Tyrosine Kinase Inhibitor; Tyrosine Phosphorylation; chemotherapy; genetic information; individualized medicine; lung tumorigenesis; nano-string; targeted treatment

In collaboration with Todd Golub at the Broad Institute, we have performed a luminex assay to quantitate tyrosine phosphorylation in tyrosine kinases. Active tyrosine kinases are also generally associated with increased overall tyrosine phosphorylation. Our assay captured more than 150 tyrosine kinases in a multiplex assay and detected the level of tyrosine phosphorylation in all these kinases using a pan-phosphotyrosine antibody (4G10). We used 150 human lung adenocarcinoma tumor tissue and corresponding adjacent normal lung tissue to profile tyrosine phosphorylation of these tyrosine kinases. We identified around 30 specific tyrosine kinases differentially phosphorylated in tumor tissue compared to adjacent normal lung. Examples of such potentially active tyrosine kinases are NTRK1, EPHA2, DDR. We then completed a nanostring assay in collaboration with Dr. Marc Ladanyi at MSKCC to measure transcript expression of majority of tyrosine kinases in a multiplex assay. Interestingly, as we had predicted, expression of several tyrosine kinases do not change at least at the level of trasnscription, however there is significant change in tyrosine phosphorylation, which is generally a surrogate of activation. In collaboration with Dr. Paul Meltzer in the NCI intramural program, we have sequenced around 1300 cancer-related genes, including the tyrosine kinases used in our luminex assay to identify specific mutations in a targeted manner. We are currently analyzing the sequencing data to identify somatic changes in the genes selected. The somatic mutation data is being correlated with the luminex data to identify possible somatic changes in kinases resulting in increased activity.

我们与 Broad 研究所的 Todd Golub 合作，进行了 luminex 测定，以定量酪氨酸激酶中的酪氨酸磷酸化。活性酪氨酸激酶通常也与整体酪氨酸磷酸化增加有关。我们的检测在多重检测中捕获了 150 多种酪氨酸激酶，并使用泛磷酸酪氨酸抗体 (4G10) 检测了所有这些激酶中的酪氨酸磷酸化水平。我们使用 150 个人类肺腺癌肿瘤组织和相应的邻近正常肺组织来分析这些酪氨酸激酶的酪氨酸磷酸化。我们鉴定出大约 30 种特定的酪氨酸激酶在肿瘤组织中与邻近的正常肺相比有差异性磷酸化。此类潜在活性酪氨酸激酶的例子有 NTRK1、EPHA2、DDR。然后，我们与 MSKCC 的 Marc Ladanyi 博士合作完成了一项纳米线测定，以在多重测定中测量大多数酪氨酸激酶的转录表达。有趣的是，正如我们所预测的，几种酪氨酸激酶的表达至少在转录水平上没有变化，但是酪氨酸磷酸化存在显着变化，这通常是激活的替代。我们与 NCI 校内项目的 Paul Meltzer 博士合作，对大约 1300 个癌症相关基因进行了测序，包括我们的 luminex 测定中使用的酪氨酸激酶，以有针对性地识别特定突变。我们目前正在分析测序数据，以确定所选基因的体细胞变化。体细胞突变数据与 luminex 数据相关联，以确定激酶中可能发生的体细胞变化，从而导致活性增加。