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Int J Stem Cell Res Ther. 2016;3(2). pii: 031. Epub 2016 May 24.

Epigenetic Loss of MLH1 Expression in Normal Human Hematopoietic Stem Cell Clones is Defined by the Promoter CpG Methylation Pattern Observed by High-Throughput Methylation Specific Sequencing.

Author information

1
Department of Pathology, Case Western Reserve University, Cleveland, OH, 44106, USA; Division of Hematology and Oncology, Case Western Reserve University, Cleveland, OH, 44106, USA; Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, OH, 44106, USA.
2
Division of Infectious Disease, Department of Medicine, Case School of Medicine and the Center for AIDS Research, Case Western Reserve University, Cleveland, OH, 44106, USA.
3
Division of Hematology and Oncology, Case Western Reserve University, Cleveland, OH, 44106, USA; Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, OH, 44106, USA.
4
Division of Hematology and Oncology, Case Western Reserve University, Cleveland, OH, 44106, USA.
5
Department of Epidemiology & Biostatistics, Case Western Reserve University, Cleveland, OH, 44106, USA.
6
Department of Mathematics, Texas State University, San Marcos, TX, 78666, USA.
7
RNA Center, Case Western Reserve University, Cleveland, OH, 44106, USA.
8
Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, OH, 44106, USA; Center for Stem Cell and Regenerative Medicine, Case Western Reserve University and University Hospitals of Cleveland, Cleveland, OH, 44106, USA.
9
Department of Pathology, Case Western Reserve University, Cleveland, OH, 44106, USA; Division of Hematology and Oncology, Case Western Reserve University, Cleveland, OH, 44106, USA; Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, OH, 44106, USA; Center for Stem Cell and Regenerative Medicine, Case Western Reserve University and University Hospitals of Cleveland, Cleveland, OH, 44106, USA; Seidman Cancer Center, University Hospitals of Cleveland, Cleveland, OH, 44106, USA.

Abstract

Normal human hematopoietic stem and progenitor cells (HPC) lose expression of MLH1, an important mismatch repair (MMR) pathway gene, with age. Loss of MMR leads to replication dependent mutational events and microsatellite instability observed in secondary acute myelogenous leukemia and other hematologic malignancies. Epigenetic CpG methylation upstream of the MLH1 promoter is a contributing factor to acquired loss of MLH1 expression in tumors of the epithelia and proximal mucosa. Using single molecule high-throughput bisulfite sequencing we have characterized the CpG methylation landscape from -938 to -337 bp upstream of the MLH1 transcriptional start site (position +0), from 30 hematopoietic colony forming cell clones (CFC) either expressing or not expressing MLH1. We identify a correlation between MLH1 promoter methylation and loss of MLH1 expression. Additionally, using the CpG site methylation frequencies obtained in this study we were able to generate a classification algorithm capable of sorting the expressing and non-expressing CFC. Thus, as has been previously described for many tumor cell types, we report for the first time a correlation between the loss of MLH1 expression and increased MLH1 promoter methylation in CFC derived from CD34+ selected hematopoietic stem and progenitor cells.

KEYWORDS:

Epigenetics; Hematopoietic stem cells; High throughput bisulfite sequencing; Mismatch repair

PMID:
27570841
PMCID:
PMC4996274

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