Metastatic protein network in BRAFV600E positive human thyroid cancers

BRAFV600E 阳性人类甲状腺癌的转移蛋白网络

• 批准号: 8386065
• 负责人: Carmelo Nucera
• 金额: $ 26.49万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-06 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8386065
• 关键词: Adhesions; BRAF gene; Binding; Binding Sites; Biological; Biological Assay; Biological Markers; Cell Line; Cell Proliferation; Cells; ChIP-seq; Clinical; Clinical Trials; DNA Binding; Data; Development; Disease; Disease Progression; Drug resistance; ECM receptor; Early Diagnosis; Endocrine; Extracellular Matrix; Extracellular Matrix Proteins; Focal Adhesion Kinase 1; Fostering; Gene Expression; Genes; Genetic Transcription; HMGB1 Protein; Human; Immunohistochemistry; In Vitro; Innovative Therapy; Integrin Binding; Integrins; Investigation; Link; Liquid substance; MAPK3 gene; Malignant Neoplasms; Malignant neoplasm of thyroid; Mass Spectrum Analysis; Modeling; Molecular; Mutation; N-terminal; Neck; Neoplasm Metastasis; Nuclear; Nuclear Translocation; Oral Administration; Papillary; Paraffin; Pathologic; Pathway interactions; Patient Monitoring; Patients; Phenotype; Phosphorylation; Phosphorylation Site; Phosphotransferases; Point Mutation; Process; Proteins; Proteomics; Recombinants; Recurrence; Refractory; Regulation; Residual Tumors; Residual state; Resistance; Sampling; Screening procedure; Sequence Analysis; Serum Response Element; Signal Transduction; Thrombospondin 1; Thyroid Gland; Thyroid carcinoma; Tissue Microarray; Tissues; Transcriptional Regulation; anaplastic thyroid cancer; base; cancer cell; cell motility; effective therapy; feeding; inhibitor/antagonist; insight; integrin-linked kinase; knock-down; melanoma; metastatic process; migration; mortality; mouse model; mutant; neoplastic cell; new therapeutic target; novel; outcome forecast; overexpression; prognostic; promoter; protein expression; receptor; small hairpin RNA; therapeutic target; thyroid neoplasm; transcription factor; tumor

DESCRIPTION (provided by applicant): A subset of patients with thyroid cancer has neck recurrences and metastases, is refractory to current treatments, and dies of the disease. BRAFV600E, the most frequent genetic alteration in both papillary (PTC) and anaplastic thyroid cancers (ATC), is implicated in progression from PTC to ATC. Our Gene Set Enrichment Analysis revealed that 7 of 17 gene sets up-regulated in BRAFV600E PTC were enriched in pro-metastatic extracellular matrix (ECM) proteins (e.g. thrombospondin-1 (TSP-1)) and receptors (integrins), including the integrin-linked kinases (i.e. focal adhesion kinase (FAK)) and the transcription factor (TF) HMGB1, which has been implicated in melanoma progression and metastasis. Knockdown of either TSP-1 or BRAFV600E inhibits cell proliferation, adhesion, migration/invasion, and metastasis in BRAFV600E-positive PTC and ATC cells and knockdown of TSP-1 significantly decreases levels of phospho(p)-ERK1/2, pFAK, and integrins. The novel selective BRAFV600E inhibitor PLX4720 shrinks tumor size in an orthotopic mouse model of ATC. However, persistence of the residual tumor and resumption of TSP-1, pERK1/2, and pFAK protein expression after 3 weeks of treatment suggest that metastatic thyroid cancer cells acquire resistance to BRAFV600E inhibition by up-regulating these proteins. Our objective is to identify BRAFV600E-dependent and -independent biomarkers for aggressive PTC by determining the essential signaling networks triggered by metastatic ECM proteins in BRAFV600E PTC cells. Specific Aim 1: To identify and assess potential biomarkers for PLX4720-resistant PTC by defining the molecular cascades by which TSP-1 stimulates ERK1/2 and FAK phosphorylation in human BRAFV600E PTC following inhibition of BRAFV600E. Preliminary data suggest that TSP-1 is associated with neck recurrence in BRAFV600E PTC. We will determine whether TSP-1 is a valid prognostic biomarker for PTC aggressiveness and establish an immunohistochemistry-based screening process suitable for clinical trials. To identify a larger panel of potential new prognostic biomarkers in BRAFV600E positive PTC, we plan to identify TSP-1 interactors. We will assess the correlation of some of these with TSP-1 expression and with clinico-pathological features of BRAFV600E positive PTC. To obtain a better understanding of BRAFV600E function in PTC progression, we will apply an unbiased proteomic analysis combined with functional assays to determine ECM protein interactions and intracellular signaling cascades in BRAFV600E PTC cells. Specific Aim 2: To explore whether HMGB1 is a biomarker for PTC aggressiveness. We will correlate HMGB1 expression with TSP-1 expression and clinico-pathological features of BRAFV600E PTC and explore whether HMGB1 regulates expression of TSP-1 or of TFs crucial for TSP-1 expression. The results of this study are likely to (i) identify new prognostic biomarkers of aggressive BRAFV600E positive PTC that can be assayed in biological fluids and PTC tissues to help monitor patients undergoing targeted therapies and enable earlier diagnosis of these thyroid cancers, and (ii) foster development of innovative therapies for PTC refractory to current treatments. PUBLIC HEALTH RELEVANCE: A subset of patients with thyroid cancer has recurrences and metastases, is refractory to current treatments, and dies of the disease. The V600E mutation in the BRAF gene is the most frequent genetic alteration in both papillary (PTC) and anaplastic thyroid cancers (ATC), and is implicated in progression of these diseases, for which early diagnosis and new therapies are needed. Results from this study (which will integrate clinical-pathologic and translational basic endocrine investigations) will enable routine assessment of new prognostic metastatic biomarkers that could be assayed in biological fluids and thyroid carcinoma tissues, and foster the development of innovative therapies for metastatic thyroid cancers that are refractory to current treatments.

描述（由申请人提供）：一部分甲状腺癌患者有颈部复发和转移，对目前的治疗无效，并死于该疾病。BRAFV600E是乳头状（PTC）和间变性甲状腺癌（ATC）中最常见的基因改变，与PTC向ATC的进展有关。我们的基因集富集分析显示，BRAFV600E PTC中上调的17个基因集中有7个富集于促转移性细胞外基质（ECM）蛋白（如血栓反应蛋白-1 (TSP-1)）和受体（整合素），包括整合素连接激酶（如局灶黏附激酶（FAK））和