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.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21CA165039-01A1
Application #
8386065
Study Section
Molecular and Cellular Endocrinology Study Section (MCE)
Program Officer
Snyderwine, Elizabeth G
Project Start
2012-07-06
Project End
2014-06-30
Budget Start
2012-07-06
Budget End
2013-06-30
Support Year
1
Fiscal Year
2012
Total Cost
$264,915
Indirect Cost
$112,665
Name
Beth Israel Deaconess Medical Center
Department
Type
DUNS #
071723621
City
Boston
State
MA
Country
United States
Zip Code
02215
Prete, Alessandro; Lo, Agnes S; Sadow, Peter M et al. (2018) Pericytes Elicit Resistance to Vemurafenib and Sorafenib Therapy in Thyroid Carcinoma via the TSP-1/TGF?1 Axis. Clin Cancer Res 24:6078-6097
Antonello, Zeus A; Hsu, Nancy; Bhasin, Manoj et al. (2017) Vemurafenib-resistance via de novo RBM genes mutations and chromosome 5 aberrations is overcome by combined therapy with palbociclib in thyroid carcinoma with BRAFV600E. Oncotarget 8:84743-84760
Daniell, Kayla; Nucera, Carmelo (2016) Effect of the micronutrient iodine in thyroid carcinoma angiogenesis. Aging (Albany NY) 8:3180-3184
Husain, Amjad; Hu, Nina; Sadow, Peter M et al. (2016) Expression of angiogenic switch, cachexia and inflammation factors at the crossroad in undifferentiated thyroid carcinoma with BRAF(V600E). Cancer Lett 380:577-85
Smith, Neal; Nucera, Carmelo (2015) Personalized therapy in patients with anaplastic thyroid cancer: targeting genetic and epigenetic alterations. J Clin Endocrinol Metab 100:35-42
Duquette, Mark; Sadow, Peter M; Husain, Amjad et al. (2015) Metastasis-associated MCL1 and P16 copy number alterations dictate resistance to vemurafenib in a BRAFV600E patient-derived papillary thyroid carcinoma preclinical model. Oncotarget 6:42445-67
Sadow, Peter M; Dias-Santagata, Dora; Zheng, Zongli et al. (2015) Identification of insertions in PTEN and TP53 in anaplastic thyroid carcinoma with angiogenic brain metastasis. Endocr Relat Cancer 22:L23-8
Kovach, Alexandra E; Nucera, Carmelo; Lam, Quynh T et al. (2015) Genomic and immunohistochemical analysis in human adrenal cortical neoplasia reveal beta-catenin mutations as potential prognostic biomarker. Discoveries (Craiova) 3:
Antonello, Z A; Nucera, C (2014) Orthotopic mouse models for the preclinical and translational study of targeted therapies against metastatic human thyroid carcinoma with BRAF(V600E) or wild-type BRAF. Oncogene 33:5397-404
Sadow, Peter M; Priolo, Carmen; Nanni, Simona et al. (2014) Role of BRAFV600E in the first preclinical model of multifocal infiltrating myopericytoma development and microenvironment. J Natl Cancer Inst 106:

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