Role of DOK family proteins in lung tumor suppression

DOK家族蛋白在抑制肺肿瘤中的作用

基本信息

批准号：
8242824
负责人：
PIER PAOLO PANDOLFI
金额：
$ 35.02万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-01 至 2015-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8242824
关键词：
ABL1 gene Adaptor Signaling Protein Affect Alleles Alveolar Biochemical Pathway Biological Assay Bone Marrow Bone Marrow Transplantation Case-Control Studies Cells Cessation of life Chronic Myeloid Leukemia Collaborations Complement Data Diagnosis EGF gene Employee Strikes Environmental Exposure Epidermal Growth Factor Receptor Epithelial Cells Exhibits Feedback Generations Genes Genetic Genetic Polymorphism Genomics Hematopoietic Hematopoietic Neoplasms Human Immune In Vitro Incidence Individual Inherited KRAS2 gene Knock-in Mouse Knock-out Knockout Mice Lung Lung Adenocarcinoma Lung Neoplasms MAP Kinase Gene Malignant Neoplasms Malignant neoplasm of lung Mediating Memorial Sloan-Kettering Cancer Center Modeling Molecular Genetics Mus Mutant Strains Mice Mutate Mutation Non-Small-Cell Lung Carcinoma Oncogenic Pathogenesis Pathway interactions Penetrance Phenotype Predisposition Prevention Protein Family Protein Tyrosine Kinase Proteins Receptor Signaling Receptors, Antigen, B-Cell Regulation Risk Factors Role Sampling Screening procedure Signal Pathway Signal Transduction Signaling Protein Smoker Smoking Somatic Mutation Stem cells Supporting Cell System Testing Tumor Suppression Tumor Suppressor Proteins Variant Work base bcr-abl Fusion Proteins cohort human FRAP1 protein in vivo knockout animal leukemia leukemogenesis loss of function lung tumorigenesis mRNA Expression macrophage mouse model mutant novel public health relevance research study response tool tumor tumor progression tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Lung cancer is responsible for the most cancer-related deaths worldwide. This proposal aims at dissecting the signaling pathways perturbed in human lung cancer, with a focus on the mechanisms of DOK-mediated lung tumor suppression. DOK ("downstream of tyrosine kinase") family proteins are signaling proteins that modulate tyrosine kinase signaling. We recently identified DOK-1, DOK-2, and DOK-3 as lung tumor suppressors, and we further hypothesize that they are negative regulators of oncogenic EGFR-RAS signaling. In our preliminary analysis, we find that Dok-1, Dok-2, and Dok-3 single, double, and triple knockout mice develop lung adenocarcinoma with a penetrance and latency dependent on the number of lost Dok alleles. Tumors from Dok null mice exhibit Akt and Erk activation, similar to models of EGFR- or KRAS-driven tumorigenesis. In human non-small cell lung cancer (NSCLC), we observe frequent genomic loss of DOK-2 with a concomitant reduction of DOK-2 mRNA expression. Strikingly, genomic loss of DOK-2 is strongly associated with EGFR mutation status. We therefore hypothesize that DOK-2 opposes lung tumorigenesis initiated by oncogenic EGFR and RAS. Moreover, we further hypothesize that variant alleles of DOK-2 (such as DOK-2L138S) could underlie lung cancer susceptibility. The purpose of this proposal is to determine the mechanisms of DOK-mediated lung tumor suppression vis-¿-vis the EGFR-RAS pathway and to ascertain the relevance of variant DOK-2 alleles and non-pulmonary cellular compartments to the pathogenesis of DOK-null lung cancer with the following specific aims: (1) To define the role of DOK-2 in EGFR- and KRAS-mutant lung cancers. To this end, we will utilize already existing EGFR and KRAS bitransgenic and Dok-2-/- mouse models to determine if Dok-2 opposes mutant EGFR or KRAS-driven lung cancer in vivo. In a preliminary analysis, we find that Dok-2 does indeed oppose lung tumorigenesis initiated by KRASG12D. (2) To define the role of DOK-2L138S in lung cancer susceptibility. The DOK-2L138S allele is present both as a somatic mutation found in human lung cancer and as a naturally occurring germline polymorphism. In vitro analysis indicates that DOK-2L138S is a loss-of-function mutant that is defective at suppressing EGF-induced RAS and ERK activity. To determine if this allele contributes to lung cancer susceptibility, we will perform a case/control study of human lung cancer, and, in parallel, generate a Dok-2L138S knock-in mouse model. (3) Evaluate the contributions of cell autonomous and non-cell autonomous mechanisms to tumor formation in Dok mutant mice. The relevance of the Dok TKO hematopoietic phenotype will be determined using a reciprocal bone marrow transplant using wild-type and TKO animals. Furthermore, generation of a lung-specific conditional Dok-2 knockout mouse model will allow unequivocal determination of the contribution of cell autonomous mechanisms to the Dok-2 knockout lung phenotype. PUBLIC HEALTH RELEVANCE: Lung cancer remains responsible for the most cancer-related deaths every year, with a growing proportion of lung cancer found in never smokers. It is therefore imperative to deconstruct the molecular genetics and the pathways perturbed in human lung cancer. Our work will define the mechanism of tumor predisposition, initiation and progression in a DOK-null scenario, in both a somatic and inherited setting, and dissect the relationship of DOK-2 loss to EGFR and RAS. This work could identify additional targets for the prevention and treatment of human lung cancer and discover genetic criteria that determine tumor phenotype, progression, and response to therapy.

描述（由适用提供）：肺癌是全世界最与癌症相关的死亡人数最多的原因。该建议旨在剖析人类肺癌中受干扰的信号通路，重点是DOK介导的肺肿瘤抑制的机制。 DOK（“酪氨酸激酶的下游”）家族蛋白是调节酪氨酸激酶信号传导的信号蛋白。我们最近将DOK-1，DOK-2和DOK-3确定为肺部肿瘤补充剂，我们进一步假设它们是致癌EGFR-RAS信号传导的负调节剂。在我们的初步分析中，我们发现DOK-1，DOK-2和DOK-3单，双敲除小鼠会发展为肺腺癌，其渗透率和潜伏期取决于丢失的DOK等位基因的数量。来自DOK NULL小鼠的肿瘤表现出AKT和ERK激活，类似于EGFR或KRAS驱动的肿瘤发生模型。在人类非小细胞肺癌（NSCLC）中，我们经常观察到DOK-2的基因组丧失，而DOK-2 mRNA表达也随之降低。令人惊讶的是，DOK-2的基因组丧失与EGFR突变状态密切相关。因此，我们假设DOK-2反对致癌EGFR和RAS引发的肺肿瘤发生。此外，我们进一步假设DOK-2的变异等位基因（例如DOK-2L138S）可能是肺癌敏感性的基础。该提案的目的是确定eGfr-ras途径DOK介导的肺肿瘤抑制的机制，并确定变异的DOK-2等位基因和非肺细胞隔室与Dok-Null Lung Cancer的发病机理与以下特定目标的相关性：为此，我们将利用已经存在的EGFR和KRAS BITTRANSGENIC和DOK-2 - / - 小鼠模型来确定DOK-2在体内是否反对突变的EGFR或KRAS驱动的肺癌。在初步分析中，我们发现DOK-2确实反对Krasg12d发起的肺肿瘤。（2）定义DOK-2L138在肺癌易感性中的作用。 DOK-2L138S等位基因既作为人类肺癌中发现的体细胞突变，也是天然存在的种系多态性。体外分析表明，DOK-2L138S是一种功能丧失突变体，在抑制EGF诱导的RAS和ERK活性方面有缺陷。为了确定该等位基因是否有助于肺癌的易感性，我们将对人类肺癌进行病例/对照研究，并同时产生DOK-2L138S敲入小鼠模型。（3）评估细胞自主和非细胞自主机制对DOK突变小鼠肿瘤形成的贡献。 DOK TKO造血表型的相关性将使用使用野生型和TKO动物的相互骨髓移植来确定。此外，生成肺特异性条件DOK-2基因敲除小鼠模型将明确确定细胞自主机制对DOK-2敲除肺表型的贡献。公共卫生相关性：每年肺癌仍然对最与癌症相关的死亡人数最多，而从未吸烟者发现的肺癌越来越多。因此，必须解构人类肺癌中的分子遗传学和途径。我们的工作将在Dok-null场景中，在体细胞和遗传环境中定义肿瘤易感性，起始和进展的机制，并剖析DOK-2损失与EGFR和RAS的关系。这项工作可以确定预防和治疗人肺癌的其他目标，并发现决定肿瘤表型，进展和对治疗反应的遗传标准。