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）：甲状腺癌患者的一部分具有颈部复发和转移，对当前治疗和死亡是难治性。 BRAFV600E，是乳头状（PTC）和型甲状腺甲状腺癌（ATC）中最常见的遗传改变，与从PTC到ATC的发展有关。我们的基因集富集分析表明，在BRAFV600E PTC中上调的17个基因中有7个富集在促逆转盘外基质基质（ECM）蛋白（例如血栓形成蛋白1（TSP-1（TSP-1））和包括整合素在内 - 连接激酶（即焦点粘附激酶（FAK））和 转录因子（TF）HMGB1与黑色素瘤进展和转移有关。 TSP-1或BRAFV600E的敲低抑制了BRAFV600E阳性PTC和ATC细胞和TSP-1的细胞增殖，粘附，迁移/侵袭和转移，而TSP-1的敲低显着降低了Phospho（P）-ERK1/2，PFAK，PFAK，PFAK和PFAK，和PFAK，以及PFAK，以及PFAK，和PFAK，以及整合素。新型的选择性BRAFV600E抑制剂PLX4720在ATC的原位小鼠模型中缩小了肿瘤的大小。但是，治疗3周后，残留肿瘤的持久性以及TSP-1，PERK1/2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质获得了对BRAFV600E抑制的抵抗力。我们的目标是通过确定由BRAFV600E PTC细胞中的转移性ECM蛋白触发的基本信号网络来鉴定侵略性PTC的BRAFV600E和非依赖性生物标志物。具体目的1：通过定义抑制BRAFV600E后，TSP-1刺激ERK1/2和FAK磷酸化的分子级联，识别和评估PLX4720耐药性PTC的潜在生物标志物。初步数据表明，TSP-1与BRAFV600E PTC中的颈部复发有关。我们将确定TSP-1是否是PTC侵略性的有效预后生物标志物，并建立基于免疫组织化学的筛查过程，适用于临床试验。为了识别BRAFV600E阳性PTC中一个更大的潜在新预后生物标志物，我们计划识别TSP-1相互作用者。我们将评估其中一些与TSP-1表达的相关性以及BRAFV600E阳性PTC的临床病理特征。为了更好地了解PTC进展中BRAFV600E功能，我们将应用无偏的蛋白质组学分析与功能测定相结合，以确定BRAFV600E PTC细胞中的ECM蛋白相互作用和细胞内信号级联。特定目的2：探索HMGB1是否是PTC侵略性的生物标志物。我们将将HMGB1的表达与TSP-1表达和BRAFV600E PTC的临床病理特征相关联，并探索HMGB1是否调节TSP-1的表达或TFS的表达对于TSP-1表达至关重要。这项研究的结果很可能（i）确定可以在生物液和PTC组织中测定的侵略性BRAFV600E阳性PTC的新的预后生物标志物，以帮助监测接受靶向疗法的患者，并能够早期诊断这些甲状腺癌的诊断，以及（ii）（ii）（ii）促进PTC难治性创新疗法的开发。

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

Effect of the micronutrient iodine in thyroid carcinoma angiogenesis.

• 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

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