LIMD2 Regulates Key Steps of Metastasis Cascade in Papillary Thyroid Cancer Cells via MAPK Crosstalk

Author(s)

Araldi, Rodrigo Pinheiro ^[1] ; de Melo, Thatiana Correa ^[2] ; Levy, Debora ^[3] ; de Souza, Dener Madeiro ^[4] ; Mauricio, Beatriz ^[5] ; Colozza-Gama, Gabriel Avelar ^[1] ; Bydlowski, Sergio Paulo ^[3] ; Peng, Hongzhuang ^[6] ; Rauscher, III, Frank J. ; Cerutti, Janete Maria ^[1]

Affiliation

^[1] Univ Fed Sao Paulo EPM, Dept Morphol & Genet, Div Genet, Genet Bases Thyroid Tumors Lab, BR-04039032 Sao Paulo, SP - Brazil

^[2] Univ Fed Integracao Latino Amer UNILA, Foz Iguacu, Programa Posgrad Biociencias, BR-85866000 Foz Do Iguacu, PR - Brazil

^[3] Univ Sao Paulo, Fac Med, Lab Histocompatibil & Cellular Immun, BR-05404000 Sao Paulo, SP - Brazil

^[4] Inst Butantan, Genet Lab, BR-05503900 Sao Paulo, SP - Brazil

^[5] Inst Butantan, Lab Cell Biol, BR-05503900 Sao Paulo, SP - Brazil

^[6] III, Wistar Inst Anat & Biol, Philadelphia, PA 19104 - USA

• Document type: Journal article
• Source: CELLS; v. 9, n. 11 NOV 2020.
• Web of Science Citations: 0

Abstract

Although papillary thyroid carcinoma (PTC) has a good prognosis, 20-90% of patients show metastasis to regional lymph nodes and 10-15% of patients show metastasis to distant sites. Metastatic disease represents the main clinical challenge that impacts survival rate. We previously showed that LIMD2 was a novel metastasis-associated gene. In this study, to interrogate the role of LIMD2 in cancer invasion and metastasis, we used CRISPR-mediated knockout (KO) of LIMD2 in PTC cells (BCPAP and TPC1). Western blot and high-content screening (HCS) analysis confirmed functional KO of LIMD2. LIMD2 KO reduced in vitro invasion and migration. Ultrastructural analyses showed that cell polarity and mitochondria function and morphology were restored in LIMD2 KO cells. To unveil the signals supervising these phenotypic changes, we employed phospho-protein array. Several members of the MAPK superfamily showed robust reduction in phosphorylation. A Venn diagram displayed the overlap of kinases with reduced phosphorylation in both cell lines and showed that they were able to initiate or sustain the epithelial-mesenchymal transition (EMT) and DNA damage checkpoint. Flow cytometry and HCS validation analyses further corroborated the phospho-protein array data. Collectively, our findings show that LIMD2 enhances phosphorylation of kinases associated with EMT and invasion. Through cooperation with different kinases, it contributes to the increased genomic instability that ultimately promotes PTC progression. (AU)

• FAPESP's process: 17/14948-7 - Study of LIMD2 role on thyroid cancer epithelial-mesenchymal transition
• Grantee: Rodrigo Pinheiro Araldi
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/06570-6 - Comprehensive whole exome, paired-end RNA and genome sequencing: new insights into genetic bases of thyroid carcinoma in pediatric and adult ages and applications in clinical practice
• Grantee: Janete Maria Cerutti
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 18/13203-0 - New insights into the prognosis of papillary thyroid microcarcinoma: mutational status analysis of BRAF V600E and whole exome sequencing
• Grantee: Gabriel Avelar Colozza Gama
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)