Comparison of genome-wide DNA methylation mapping between CD34+CD38-ALDHhigh and CD34+CD38-ALDHint cell subsets isolated from acute myeloid leukemia patients

• Grant number: 14/23651-0
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: March 01, 2015
• End date: May 31, 2015
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Eduardo Magalhães Rego
• Grantee: Mariana Tereza de Lira Benício
• Supervisor: Daniel Diniz de Carvalho
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Institution abroad: University of Toronto (U of T), Canada
• Associated to the scholarship: 11/17112-0 - Assessment of signaling pathways in stem-cells of de novo acute myeloid leukemia, BP.DR

Abstract

Despite the great variability of genetic and epigenetic alterations found in AML, many of them often target signal transduction pathways, leading to dysregulation of downstream transcriptional effectors and widespread gene expression changes. There is growing evidence that signal transduction pathways dysregulation originate in early progenitor and possibly stem cell compartments, since many of the cells exhibiting aberrant signaling also show cell surface attributes of a stem/progenitor cell phenotype. Therefore, finding pathways in these cells that could be therapeutically targeted would be an efficient approach to cure AML. We have shown that there are different signaling patterns between putative normal and leukemic stem cells, derived from the same patient, but the underlying mechanisms remain unknown. Since there are many evidences that these cell subsets originate from a common precursor, we hypothesized that there may be a role for epigenetics in this process. This emerges the possibility of mapping these epigenetic modifications in order to identify signatures that predict resistance or sensitivity to chemotherapy, which could also be correlated to data generated by cell signaling studies. Base-pair resolution genome-wide mapping of DNA methylation will be generated using enhanced reduced representation bisulfite sequencing (ERRBS), a bisulfite-based protocol that enriches GC-rich parts of the genome, thereby reducing costs while still capturing most of the genomic regions we are interested in (CpG islands and shores). This method is suitable to use with starting material as little as 500 cells, a necessary condition for these highly purified cells. We will compare the DNA methylation maps among CD34+CD38-ALDHhigh and CD34+CD38-ALDHint cell subsets isolated from acute myeloid leukemia patients, and CD34+CD38-ALDHhigh cells isolated from normal donors, in order to get more insight into the crosstalk between epigenetics and cell signaling, and to determine if DNA methylation distributes into specific patterns between the previously mentioned cell subsets. (AU)