Disabled-2 in the metabolic regulation of oncopathways

Disabled-2 在肿瘤途径代谢调节中的作用

• 批准号: 10578523
• 负责人: Benny Abraham Kaipparettu
• 金额: $ 8万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-09 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10578523
• 关键词: Affect; Binding; Breast Cancer Cell; Breast Cancer cell line; Breast cancer metastasis; Cell Nucleus; Cells; Cellular biology; Characteristics; Clinical; Communication; Core Facility; DNA Methylation; Data; Dependence; Disabled Persons; Down-Regulation; Drug resistance; Enzymes; Epigenetic Process; Excision; Fatty Acids; Focal Adhesion Kinase 1; Funding; Genes; Genetic Transcription; Glucose; Glutamine; Histone Deacetylase; Histone Deacetylase Inhibitor; Laboratories; Leptin; Malignant Neoplasms; Mechanics; Mediating; Messenger RNA; Metabolic; Metabolic Activation; Mitochondria; Modeling; Modification; Molecular; Molecular Biology; Mus; Mutation; Neoplasm Metastasis; Nuclear; Outcome; Oxidative Phosphorylation; PTK2 gene; Pancreas; Pathway interactions; Patient-derived xenograft models of breast cancer; Phosphorylation; Phosphotransferases; Play; Post-Translational Regulation; Prognosis; Protein Dephosphorylation; Protein Kinase; Proteins; Proto-Oncogenes; Publications; Publishing; Regulation; Reporting; Research; Research Personnel; Resources; Role; SH3 Domains; SRC gene; Secure; Signal Pathway; Signal Transduction; Squamous cell carcinoma; Subgroup; Transcriptional Regulation; Tumor Suppressor Genes; Tumor Suppressor Proteins; Tumor Tissue; United States National Institutes of Health; Validation; Xenograft procedure; cancer subtypes; clinically significant; epigenetic regulation; experience; fatty acid oxidation; histone modification; hormone receptor-positive; in vivo; inhibitor; knock-down; malignant breast neoplasm; metabolomics; ovarian neoplasm; oxidation; patient derived xenograft model; predict clinical outcome; promoter; small hairpin RNA; src-Family Kinases; translational study; triple-negative invasive breast carcinoma; tumor; tumor progression; tumor xenograft; tumorigenesis; v-src Oncogenes

Abstract: Compared to hormone receptor (HR) positive breast cancer (BC), basal or triple negative BC (TNBC) suffers a poor prognosis and limited treatment options because of the lack of understanding of its driver signaling pathways. Mitochondrial retrograde signaling is a pathway of communication from mitochondria to the nucleus. We have previously published that TNBC has energy dependency to mitochondrial FA β-oxidation (FAO). FAO induces tumor and metastatic potential of TNBC. Src oncopathway is one of the most frequently upregulated pathways in TNBC and a critical player of TNBC metastasis. Like most protein kinases, Src family kinases (SFKs) require phosphorylation within a segment of the kinase for its full catalytic activity. Src is mainly phosphorylated (pSrc) at either Y419 (active state) or Y530 (inactive state). Compared to most BC subgroups, pSrc (Y419) is significantly upregulated in TNBC subgroups. However, how pSrc oncopathway is activated in TNBC was largely unknown. In this context, our lab made a breakthrough discovery that in TNBC, FAO regulates the phosphorylation of Src at Y419 without any major impact on the phosphorylation of Y530. Though we have shown that FAO inhibition decreases the pSrc (Y419), so far, the mechanism of this critical regulation is not fully understood. Only very fewer proteins have been proposed for the removal of phosphorylation from pSrc (Y419). Disabled-2 (DAB2), is known to bind at SH3 domain of Src and directly suppress the Y419 phosphorylation. This dephosphorylation results in the inactivation for Src without affecting the phosphorylation at Y530. Previous studies have suggested that the epigenetic transcriptional downregulation of DAB2 is a major determinant of cancer progression and clinical outcome. FA and FAO metabolites are known to epigenetically modify nuclear genes. Our strong preliminary data using different models suggest that the FAO downregulate the DAB2 mRNA in TNBC. Moreover, HDAC inhibitors activate DAB2 mRNA in TNBC cells. All these suggest that FAO may be epigenetically regulating DAB2. Thus, in this project we will evaluate how FAO-mediated retrograde signaling epigenetically modify the tumor suppressor DAB2. We will also analyze if such regulation of DAB2 is critical in the FAO-mediated activation of Src oncopathway in TNBC. In Aim-1, FAO-mediated transcriptional regulation of DAB2 will be confirmed using TNBC cell lines and already available tumor tissues collected after long-term treatment with FAO inhibitors in mice bearing TNBC patient-derived xenograft (PDX) models. We also have TNBC cells after stably knocking down FAO rate-limiting enzymes. Aim-2 will analyze the epigenetic modifications of DAB2 promoter after metabolic modulation. Overall, this will be one of the first mechanical studies in TNBC to show how the mitochondrial retrograde regulation modulate a tumor suppressor gene to control a major oncopathway via its post-translational regulation. The investigator team is well experienced in the proposed studies and supported by other core facilities. Strong preliminary data and previous publications support the feasibility of this project.

摘要：与激素受体（HR）阳性乳腺癌（BC）相比，基础或三阴性BC（TNBC） 由于缺乏对其驱动信号的了解， 途径。线粒体逆行信号传导是从线粒体到细胞核的通信途径。 我们之前已经发表了TNBC对线粒体FA β-氧化（FAO）具有能量依赖性。粮农组织 诱导TNBC的肿瘤和转移潜力。Src oncopathway是最常上调的一种 在TNBC中的通路和TNBC转移的关键球员。与大多数蛋白激酶一样，Src家族激酶（SFK） 需要在激酶的一个片段内磷酸化以获得其全部催化活性。Src主要被磷酸化 （pSrc）在Y 419（活性状态）或Y 530（非活性状态）处。与大多数BC亚组相比，pSrc（Y 419）是 在TNBC亚组中显著上调。然而，在TNBC中pSrc肿瘤通路如何被激活在很大程度上是未知的。 未知在此背景下，我们实验室取得了突破性的发现，在TNBC中，FAO调节了 在Y 419处的Src的磷酸化没有对Y 530的磷酸化的任何主要影响。虽然我们 表明FAO抑制降低了pSrc（Y 419），到目前为止，这一关键调节的机制还不完全 明白只有很少的蛋白质被提出用于从pSrc（Y 419）中去除磷酸化。 DAB 2与Src的SH 3结构域结合，直接抑制Y 419的磷酸化。这 去磷酸化导致Src失活，而不影响Y 530处的磷酸化。先前 研究表明，DAB 2的表观遗传转录下调是DAB 2的主要决定因素。 癌症进展和临床结果。已知FA和FAO代谢物可表观遗传修饰细胞核 基因.我们使用不同模型的强有力的初步数据表明，FAO下调DAB 2 mRNA， 在TNBC。此外，HDAC抑制剂激活TNBC细胞中的DAB 2 mRNA。所有这些都表明，粮农组织可能是 表观遗传调节DAB 2。因此，在这个项目中，我们将评估粮农组织介导的逆行信号传导， 表观遗传修饰肿瘤抑制因子DAB 2。我们还将分析DAB 2的这种调节是否在以下方面至关重要： FAO介导的TNBC中Src oncopathway的激活。在Aim-1中，FAO介导的转录调节 DAB 2将使用TNBC细胞系和在长期化疗后收集的已经可用的肿瘤组织进行确认。 在携带TNBC患者来源的异种移植物（PDX）模型的小鼠中用FAO抑制剂治疗。我们也有 稳定敲除FAO限速酶后的TNBC细胞。Aim-2将分析表观遗传 代谢调节后DAB 2启动子的修饰。总的来说，这将是第一个机械 在TNBC中的研究显示线粒体逆行调节如何调节肿瘤抑制基因， 通过其翻译后调节来控制主要的肿瘤通路。调查员团队在以下方面经验丰富： 建议的研究和其他核心设施的支持。强有力的初步数据和以前的出版物 支持该项目的可行性。