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: 8507182
• 负责人: Carmelo Nucera
• 金额: $ 14.23万
• 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/8507182
• 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; Prognostic Marker; Proteins; Proteomics; Recombinants; Recurrence; Refractory; Regulation; Residual Tumors; Residual state; Resistance; Sampling; 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; screening; 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.

描述（由申请人提供）：甲状腺癌患者的一个子集有颈部复发和转移，对目前的治疗难治，并死于这种疾病。BRAFV 600 E是乳头状癌（PTC）和甲状腺间变性癌（ATC）中最常见的遗传变异，与PTC向ATC的进展有关。我们的基因集富集分析显示，在BRAFV 600 E PTC中上调的17个基因集中有7个富含促转移细胞外基质（ECM）蛋白（例如血小板反应蛋白-1（TSP-1））和受体（整联蛋白），包括整联蛋白连接的激酶（即粘着斑激酶（FAK））， 转录因子（TF）HMGB 1，其与黑色素瘤进展和转移有关。TSP-1或BRAFV 600 E的敲低抑制BRAFV 600 E阳性PTC和ATC细胞中的细胞增殖、粘附、迁移/侵袭和转移，并且TSP-1的敲低显著降低磷酸（p）-ERK 1/2、pFAK和整联蛋白的水平。新型选择性BRAFV 600 E抑制剂PLX 4720在ATC原位小鼠模型中缩小肿瘤大小。然而，治疗3周后残留肿瘤的持续存在和TSP-1、pERK 1/2和pFAK蛋白表达的恢复表明，转移性甲状腺癌细胞通过上调这些蛋白获得对BRAFV 600 E抑制的抗性。我们的目标是通过确定BRAFV 600 E PTC细胞中转移性ECM蛋白触发的基本信号传导网络，鉴定侵袭性PTC的BRAFV 600 E依赖性和非依赖性生物标志物。具体目标1：通过定义抑制BRAFV 600 E后TSP-1刺激人BRAFV 600 E PTC中ERK 1/2和FAK磷酸化的分子级联，鉴定和评估PLX 4720耐药PTC的潜在生物标志物。初步数据表明，TSP-1与BRAFV 600 E PTC的颈部复发相关。我们将确定TSP-1是否是PTC侵袭性的有效预后生物标志物，并建立一个适合临床试验的基于免疫化学的筛查过程。为了在BRAFV 600 E阳性PTC中鉴定更多潜在的新预后生物标志物，我们计划鉴定TSP-1相互作用物。我们将评估其中一些与TSP-1表达和BRAFV 600 E阳性PTC的临床病理特征的相关性。为了更好地了解BRAFV 600 E在PTC进展中的功能，我们将应用无偏蛋白质组学分析结合功能测定来确定BRAFV 600 E PTC细胞中ECM蛋白相互作用和细胞内信号级联。具体目的2：探讨HMGB 1是否是PTC侵袭性的生物标志物。我们将HMGB 1表达与TSP-1表达和BRAFV 600 E PTC的临床病理特征相关联，并探讨HMGB 1是否调节TSP-1或对TSP-1表达至关重要的TF的表达。这项研究的结果可能（i）确定侵袭性BRAFV 600 E阳性PTC的新预后生物标志物，这些生物标志物可以在生物液体和PTC组织中进行测定，以帮助监测接受靶向治疗的患者，并使这些甲状腺癌的早期诊断成为可能，以及（ii）促进对目前治疗难治性PTC的创新疗法的开发。

项目成果

Orthotopic mouse models for the preclinical and translational study of targeted therapies against metastatic human thyroid carcinoma with BRAF(V600E) or wild-type BRAF.

• DOI: 10.1038/onc.2013.544
• 发表时间: 2014-11-20
• 期刊: Oncogene
• 影响因子: 8
• 作者: Antonello ZA;Nucera C
• 通讯作者: Nucera C

Genomic and immunohistochemical analysis in human adrenal cortical neoplasia reveal beta-catenin mutations as potential prognostic biomarker.

人类肾上腺皮质肿瘤的基因组和免疫组织化学分析揭示β-连环蛋白突变是潜在的预后生物标志物。

• DOI: 10.15190/d.2015.32
• 发表时间: 2015
• 期刊: Discoveries (Craiova, Romania)
• 作者: Kovach,AlexandraE;Nucera,Carmelo;Lam,QuynhT;Nguyen,Ahnthu;Dias-Santagata,Dora;Sadow,PeterM
• 通讯作者: Sadow,PeterM

Pericytes Elicit Resistance to Vemurafenib and Sorafenib Therapy in Thyroid Carcinoma via the TSP-1/TGFβ1 Axis.

• DOI: 10.1158/1078-0432.ccr-18-0693
• 发表时间: 2018-12-01
• 期刊: Clinical cancer research : an official journal of the American Association for Cancer Research
• 作者: Prete A;Lo AS;Sadow PM;Bhasin SS;Antonello ZA;Vodopivec DM;Ullas S;Sims JN;Clohessy J;Dvorak AM;Sciuto T;Bhasin M;Murphy-Ullrich JE;Lawler J;Karumanchi SA;Nucera C
• 通讯作者: Nucera C

FOXM1 is a molecular determinant of the mitogenic and invasive phenotype of anaplastic thyroid carcinoma.

• DOI: 10.1530/erc-12-0031
• 发表时间: 2012-10
• 期刊: Endocrine-related cancer
• 影响因子: 3.9
• 作者: Bellelli R;Castellone MD;Garcia-Rostan G;Ugolini C;Nucera C;Sadow PM;Nappi TC;Salerno P;Cantisani MC;Basolo F;Gago TA;Salvatore G;Santoro M
• 通讯作者: Santoro M

Effect of the micronutrient iodine in thyroid carcinoma angiogenesis.

微量营养素碘在甲状腺癌血管生成中的影响。

• DOI: 10.18632/aging.101143
• 发表时间: 2016-12-20
• 期刊: Aging
• 作者: Daniell K;Nucera C
• 通讯作者: Nucera C