Identification of the DNA hydroxymethylation profile during melanoma progression

• Grant number: 14/10349-3
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: August 01, 2014
• End date: February 29, 2020
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Miriam Galvonas Jasiulionis
• Grantee: Guilherme Burgarelli Leite
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated scholarship(s): 18/15020-0 - Identification of the genomic DNA hydroxymethylation profile during the melanoma progression, BE.EP.DD

Abstract

Tet proteins are 2-oxoglutarate (2OG) and Fe(II) dependent dioxygenases that catalyse the hydroxylation of 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC) in DNA, acting as an intemediate product either during active or passive demethylation, contributing with the DNA methylation dinamics (Koh et al., 2011; Tahiliani et al., 2009). The DNA methylation patterns are altered during the malign transformation of melanocytes, affecting its gene expression. Tet1 and Tet2 present different preferred genomic sites in which they deposit 5hmC (Huang et al., 2013). Utilizing a linear model of melanoma progression, with lineages corresponding to non-tumorigenics melanocytes (melan-a), pre-malign melanocytes (4C), non-metastatic melanoma cells (4C11-) and metastatic melanoma (4C11+), we intend to do a full genome mapping of 5hmC content in these lineages and, correlating with prior transcriptome data, characterize specific genes which expression are affected by this mark during melanoma progression. The selected genes will be validated and investigated in human lineages of metastatic melanoma and tumor specimens, and also in human model lineages of melanoma progression. The expression data of those genes will be related, if possible, with gene expression and protein expression data of Tet1 and Tet2. Furthermore, functional activity assays will be performed using silenced lineages for the genes Tet1 or Tet2, aiming to understand the biological role of these proteins. This study will allow the identification of specific genomic sites that contain 5hmC and characterize epigenetic altered genes during melanoma progression. (AU)