Characterization of AMH as part of a novel TGF-beta signaling loop

AMH 作为新型 TGF-β 信号环路一部分的表征

基本信息

批准号：
9319636
负责人：
Tim Nico Beck
金额：
$ 3.86万
依托单位：
DREXEL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-01 至 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9319636
关键词：
A549 Address Adenocarcinoma Adverse effects Alpha Cell Anti-Mullerian Hormone Receptor Type II Antibodies Autocrine Communication Automobile Driving Bioinformatics Biological Assay Biological Markers Cancer cell line Cell Line Cell Surface Receptors Cell surface Cells Cellular Morphology Chimeric Proteins Client Clinical Clinical Management Clinical Trials Communities Coupled Data Databases Development Diagnosis Drug resistance ERBB2 gene Elements Epithelial Generations Genes Goals Growth H1299 HSP 90 inhibition Heat-Shock Proteins 90 Histopathology Human Immunofluorescence Immunologic In Vitro Incidence Investigation KRAS2 gene Laboratories Libraries Ligands Link Malignant Neoplasms Malignant neoplasm of lung Mesenchymal Molecular Chaperones Mutate Neoplasm Metastasis Non-Small-Cell Lung Carcinoma Outcome Pathogenesis Pathway interactions Patients Phosphotransferases Play Progression-Free Survivals Proteins Radiation Regimen Reporter Resistance Reverse Transcriptase Polymerase Chain Reaction Role Sampling Signal Pathway Signal Transduction Signaling Protein Small Interfering RNA Sorting - Cell Movement Staining method Stains Structure of parenchyma of lung Survival Rate TGFBR1 gene Testing Therapeutic Time Tissue Microarray Tissue Sample Tissues Transforming Growth Factor beta Transforming Growth Factors Work Xenograft procedure base cancer cell cancer therapy cancer type chemotherapy design epithelial to mesenchymal transition experimental study female reproductive system genome sequencing improved in vitro testing in vivo in vivo imaging inhibitor/antagonist interest killings knock-down male member mouse model mullerian-inhibiting hormone mutant novel novel therapeutics outcome forecast overexpression pre-clinical public health relevance receptor screening small hairpin RNA support network synergism targeted treatment therapeutic evaluation transcriptome sequencing tumor tumorigenesis whole genome

项目摘要

Project Summary/Abstract. Lung cancer kills approximately 160,000 people in the US annually, making it the deadliest type of cancer, with a 5-year survival rate of only 15.9% of diagnosed cases. As a chaperone protein, heat shock protein 90 (HSP90) is essential to support activity of many proteins involved in oncogenesis, and its overexpression correlates with worse outcomes. Although early HSP90 inhibitors were not clinically successful due to toxic side effects, second-generation compounds, such as ganetespib, are showing considerable promise in late stage clinical trials, including some for lung cancer. Following extensive preliminary short interfering (si)RNA screening, I found that depletion of anti-Müllerian hormone (AMH) and the AMH type II receptor (AMHRII) significantly sensitized three KRAS-mutated lung cancer cell lines to ganetespib. Importantly, AMH is a member of the transforming growth factor (TGF)-β superfamily and relates to TGF-β signaling by sharing parts of an effector cascade. TGF-β signaling is frequently involved in lung cancer proliferation, survival and metastasis, the last being the deadliest aspect of this malignancy. TGF-β has been studied extensively, but aspects of its function in aggressive cancer remain opaque. Our preliminary data suggests a novel autocrine-signaling loop involving AMH and AMHRII, representing the first time these proteins have been identified as functioning in a non-gonadal tissue cancer context. Intriguingly, Taipale et al. recently discovered that AMHRII stability is most dependent on HSP90 out of 314 tested kinases, while online databases and my own investigations have established significant expression of AMH and AMHRII in a subset of lung cancer cell lines as well as in many primary lung cancers. Thus, we propose that AMH and AMHRII play a critical role in a subpopulation of lung cancers, and may be exploitable targets to potentiate the efficacy of current and novel therapies or at minimum be utilized as biomarkers. Our broad, long-term objectives are to further elucidate the role TGF-β plays in cancer proliferation and metastasis and how this information can be used to improve the status quo of patients afflicted with lung cancer. Our three aims will better delineate AMH as a pro-survival ligand linked to TGF-β signaling and explore the translational possibilities of our findings. For our first aim, we will use siRNA and overexpression experiments to probe interactions of AMH and the TGF-β effector cascades and explore links of AMH to epithelial-mesenchymal transition. The second aim will use in vitro testing of combinations of ganetespib with AMHIIR-targeting antibodies or an AMH-targeting fusion protein, in the context of altered AMH function, and xenograft mouse models will be used to study the effects of short hairpin RNA (shRNA) knockdown of AMH in combination with ganetespib treatment in vivo. Lastly, we will stain tissue microarrays of human lung cancer for AMH, AMHRII, HSP90 and several other proteins as part of a preclinical assessment.

项目摘要/摘要。肺癌每年在美国夺去约 16 万人的生命，使其成为最致命的癌症类型，确诊病例的 5 年生存率仅为 15.9%。作为伴侣蛋白，热休克蛋白 90 (HSP90) 对于支持至关重要。尽管早期的 HSP90 抑制剂由于毒副作用而在临床上并不成功，但第二代化合物，诸如 ganetespib 等药物在后期临床试验中显示出相当大的前景，其中包括一些针对肺癌的临床试验。经过广泛的初步短干扰 (si)RNA 筛选，我发现抗苗勒氏管激素 (AMH) 和 AMH II 型受体的消耗。 AMHRII) 使三种 KRAS 突变的肺癌细胞系对 ganetespib 显着敏感。重要的是，AMH 是转化生长因子 (TGF)-β 超家族的成员，与相关。 TGF-β 信号传导通过共享效应子级联的一部分经常参与肺癌的增殖、存活和转移，最后一个是 TGF-β 最为致命的方面已得到广泛研究。 Taipale 等人有趣的是，我们的初步数据表明，AMH 和 AMHRII 涉及一种新的自分泌信号环路，这代表着这些蛋白质首次在非性腺组织癌症中发挥作用。最近发现，在 314 种测试的激酶中，AMHRII 的稳定性最依赖于 HSP90，而在线数据库和我自己的研究已经确定了 AMH 和 AMHRII 在肺的一个子集中的显着表达因此，我们认为 AMH 和 AMHRII 在肺癌亚群中发挥着关键作用，并且可能是增强当前和新疗法疗效或至少被利用的靶标。作为生物标志物。我们广泛的长期目标是进一步阐明 TGF-β 在癌症增殖和转移中的作用，以及如何利用这些信息来改善肺癌患者的现状。我们的三个目标将更好地将 AMH 描述为一种治疗方法。与 TGF-β 信号传导相关的促生存配体，并探索我们的研究结果的转化可能性。对于我们的第一个目标，我们将使用 siRNA 和过表达实验来探测 AMH 和 TGF-β 效应子的相互作用。第二个目标是在改变的背景下使用 ganetespib 与 AMHIIR 靶向抗体或 AMH 靶向融合蛋白的组合进行体外测试。 AMH 功能和异种移植小鼠模型将用于研究 AMH 短发夹 RNA (shRNA) 敲低与体内 Ganetespib 治疗相结合的效果。最后，我们将对人肺癌组织微阵列进行 AMH、AMHRII、HSP90 和 HSP90 染色。作为临床前评估的一部分的其他几种蛋白质。