Understanding the impact that tumor representative oxygen tension has on phosphotyrosine-dependent signaling networks in solid tumors

了解肿瘤代表性氧张力对实体瘤中磷酸酪氨酸依赖性信号网络的影响

• 批准号: 10303523
• 负责人: Benjamin Jacob Mayro
• 金额: $ 3.95万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10303523
• 关键词: 2-tyrosine; ABL1 gene; ABL2 gene; Animal Model; Biological; Brain; Cell Hypoxia; Cells; Cellular biology; Characteristics; Clinical Trials; Dasatinib; Data; Dependence; Development; Disseminated Malignant Neoplasm; Doctor of Philosophy; Environment; FDA approved; Fellowship; Foundations; Future; Genetic Transcription; Gleevec; HSF1; Hypoxia; Impairment; In Vitro; Intervention; Investigation; Luciferases; Lung Adenocarcinoma; Malignant Neoplasms; Manuscripts; Mass Spectrum Analysis; Mediating; Metastatic malignant neoplasm to brain; Modality; Nature; Neoplasm Metastasis; Oxides; Oxygen; Patient-Focused Outcomes; Patients; Pharmacology; Phosphoric Monoester Hydrolases; Phosphotransferases; Phosphotyrosine; Play; Postdoctoral Fellow; Principal Investigator; Protein Tyrosine Kinase; Protein Tyrosine Phosphatase; Proteins; Proto-Oncogene Proteins c-abl; Reporter; Reporting; Research; Research Project Grants; Resistance; Role; Sampling; Signal Pathway; Signal Transduction; Solid Neoplasm; Technology; Therapeutic; Time; Training; Translating; Trastuzumab; Tumor-Associated Process; Tyrosine Kinase Inhibitor; Tyrosine Phosphorylation; Work; anticancer research; autocrine; base; brain parenchyma; cancer cell; cancer therapy; career; effective therapy; efficacious treatment; high-throughput drug screening; improved; in vivo; inhibitor/antagonist; leukemia; lung cancer cell; lung colonization; molecular targeted therapies; oxidation; pre-clinical; pre-doctoral; preclinical study; programs; response; small molecule; targeted treatment; trait; transcription factor; tumor; tumor hypoxia; tumor microenvironment; tumor progression; ubiquitin-protein ligase

PROJECT SUMMARY/ ABSTRACT The perturbation of phospho-tyrosine mediated signaling networks is an essential occurrence during the multistep process of tumor development and progression. As a result, the components of these phospho-tyrosine signaling networks, especially tyrosine kinases, have been shown to be a key reservoir of actionable molecular targets for the treatment of cancer. In recent years, it has been revealed that the tumor microenvironment plays a critical role in modulating the signaling pathways that govern tumor progression and metastasis. The features of a tumor's microenvironment have been shown to produce unique sensitivities and resistances to different treatment modalities. One major aspect of the tumor microenvironment which is often overlooked in preclinical studies is oxygen tension. This proposal seeks to understand the impact that oxygen tension has on phosphotyrosine-dependent signaling networks in solid tumors, and how the resultant vulnerabilities can be targeted to improve patient outcome. In Aim 1.1 (prior studies), we sought to identify alterations in signaling networks that occur when lung cancer cells colonize the brain, a hypoxic environment. We showed that brain- metastatic lung cancer cells elevate and have an increased dependence on a non-canonical HSF1-E2F transcriptional program for survival. Importantly, we identified that this transcriptional program is targetable through treatment with allosteric ABL2 tyrosine kinase inhibitors. In Aim 1.2 (proposed studies), using a small molecule screen, I have identified previously unrecognized modulators of the cellular response to hypoxia, a tumor microenvironment feature associated with increased metastasis and lower overall survival in patients with solid tumors. The top uncharacterized hit was the FDA-approved ABL1/2 tyrosine kinase inhibitor Dasatinib and my preliminary investigation has shown that the ABL kinases are critical regulators of HIF-1α protein stability. I will continue mechanistic investigation of the ABL- HIF-1α axis in vitro and in vivo. Finally, in Aim 2 (post-doctoral studies), I will focus on understanding the impact that tumor representative- oxygen tension has on protein tyrosine phosphatase activity. Extensive investigation has demonstrated that tumor hypoxia induces activation of phospho-tyrosine signaling networks, but current work has almost exclusively focused on the role of tyrosine kinases. I show that hypoxia induces inhibitory oxidation of protein tyrosine phosphatases (PTPs). Using mass- spectrometry based approaches, I will identify the oxidized- PTP landscape (ox-PTPome) of tumor samples and cancer cells at oxygen levels observed in tumors. Further, since PTPs restrain cellular signaling, I will employ high-throughput drug screening technologies in vitro to identify emergent sensitivities due to the loss of PTP activity that would not have been captured in the numerous normoxically (tumor-unrepresentative oxygen level) performed screens. Overall, the focus of my career is to understand how the different characteristics of the tumor microenvironment, such as hypoxia, modulates the signaling networks co-opted by cancer cells and translate this to the identification of biological mechanisms that may be amenable to therapeutic exploitation.

项目总结/摘要 磷酸酪氨酸介导的信号网络的扰动是在细胞周期中必不可少的。 肿瘤的发生和发展是一个多步骤的过程。因此，这些磷酸酪氨酸的组分 信号传导网络，特别是酪氨酸激酶，已被证明是可操作分子的关键库， 治疗癌症的靶点。近年来，研究表明，肿瘤微环境在肿瘤发生发展中起着重要作用。 在调节控制肿瘤进展和转移的信号通路中起关键作用。的特征 已经证明，肿瘤微环境的变化会对不同的肿瘤细胞产生独特的敏感性和抵抗力。 治疗方式。肿瘤微环境的一个主要方面，在临床前研究中经常被忽视， 研究的是氧张力。该提案旨在了解氧气紧张对 磷酸酪氨酸依赖的信号网络在实体瘤，以及如何产生的漏洞可以 旨在改善患者预后。在目标1.1（先前的研究）中，我们试图识别信号转导的改变， 当肺癌细胞在缺氧环境中的大脑中定居时，会出现这种网络。我们展示了大脑- 转移性肺癌细胞对非典型HSF 1-E2 F的依赖性增加 生存的转录程序。重要的是，我们发现这个转录程序是有针对性的， 通过用别构ABL 2酪氨酸激酶抑制剂治疗。在目标1.2（拟议研究）中，使用小 分子筛选，我已经确定了以前未被认识到的细胞缺氧反应的调节剂， 肿瘤微环境特征与肿瘤转移增加和总生存率降低相关 实体瘤最热门的非特征性药物是FDA批准的ABL 1/2酪氨酸激酶抑制剂达沙替尼， 我的初步研究表明，ABL激酶是HIF-1α蛋白稳定性的关键调节因子。我 将继续在体外和体内研究ABL-HIF-1α轴的机制。最后，在目标2（博士后 研究），我将专注于了解肿瘤代表-氧张力对蛋白质的影响 酪氨酸磷酸酶活性广泛的研究表明，肿瘤缺氧诱导活化 磷酸酪氨酸信号网络，但目前的工作几乎完全集中在酪氨酸的作用， 激酶。我表明，缺氧诱导抑制氧化的蛋白酪氨酸磷酸酶（PTPs）。使用mass- 基于光谱的方法，我将确定肿瘤样品的氧化- PTP景观（ox-PTPome）， 在肿瘤中观察到的氧水平的癌细胞。此外，由于PTP抑制细胞信号传导，我将采用 体外高通量药物筛选技术，以确定由于PTP丢失而引起的紧急敏感性 在许多正常氧（肿瘤-非代表性氧水平）下不会捕获的活性 进行筛选。总的来说，我职业生涯的重点是了解肿瘤的不同特征 微环境，如缺氧，调节由癌细胞选择的信号网络，并翻译 这有助于确定可能适合于治疗开发的生物学机制。