Journal Article

Globin gene expression in correlation with G protein-related genes during erythroid differentiation

Schechter, Alan N, Čokić, Vladan P, Smith, Reginald D, Noguchi, Constance T, Puri, Raj K, Biancotto, Angélique

2013

Overview

Background The guanine nucleotide binding protein (G protein)-coupled receptors (GPCRs) regulate cell growth, proliferation and differentiation. G proteins are also implicated in erythroid differentiation, and some of them are expressed principally in hematopoietic cells. GPCRs-linked NO/cGMP and p38 MAPK signaling pathways already demonstrated potency for globin gene stimulation. By analyzing erythroid progenitors, derived from hematopoietic cells through in vitro ontogeny, our study intends to determine early markers and signaling pathways of globin gene regulation and their relation to GPCR expression. Results Human hematopoietic CD34 + progenitors are isolated from fetal liver (FL), cord blood (CB), adult bone marrow (BM), peripheral blood (PB) and G-CSF stimulated mobilized PB (mPB), and then differentiated in vitro into erythroid progenitors. We find that growth capacity is most abundant in FL- and CB-derived erythroid cells. The erythroid progenitor cells are sorted as 100% CD71 + , but we did not find statistical significance in the variations of CD34, CD36 and GlyA antigens and that confirms similarity in maturation of studied ontogenic periods. During ontogeny, beta-globin gene expression reaches maximum levels in cells of adult blood origin (176 fmol/μg), while gamma-globin gene expression is consistently up-regulated in CB-derived cells (60 fmol/μg). During gamma-globin induction by hydroxycarbamide, we identify stimulated GPCRs ( PTGDR, PTGER1 ) and GPCRs-coupled genes known to be activated via the cAMP/PKA ( ADIPOQ ), MAPK pathway ( JUN ) and NO/cGMP ( PRPF18 ) signaling pathways. During ontogeny, GPR45 and ARRDC1 genes have the most prominent expression in FL-derived erythroid progenitor cells, GNL3 and GRP65 genes in CB-derived cells (high gamma-globin gene expression), GPR110 and GNG10 in BM-derived cells, GPR89C and GPR172A in PB-derived cells, and GPR44 and GNAQ genes in mPB-derived cells (high beta-globin gene expression). Conclusions These results demonstrate the concomitant activity of GPCR-coupled genes and related signaling pathways during erythropoietic stimulation of globin genes. In accordance with previous reports, the stimulation of GPCRs supports the postulated connection between cAMP/PKA and NO/cGMP pathways in activation of γ-globin expression, via JUN and p38 MAPK signaling.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V
• Subject: Analysis / Animal Genetics and Genomics / beta-Globins - genetics / beta-Globins - metabolism / Biomedical and Life Sciences / Biotechnology industry / Bone marrow / Cell Differentiation / Cell growth / Cell Proliferation / Diabetes / Erythrocytes / Erythroid Precursor Cells - cytology / Erythroid Precursor Cells - metabolism / Erythropoiesis - genetics / Fetal Blood - cytology / Fetal Blood - metabolism / Flow cytometry / G proteins / gamma-Globins - genetics / gamma-Globins - metabolism / Gene expression / Gene Expression Regulation, Developmental / Genes / Genetic aspects / Genetic research / Genomics / Globin genes / Granulocyte Colony-Stimulating Factor - genetics / GTP-Binding Proteins - genetics / GTP-Binding Proteins - metabolism / Human and rodent genomics / Humans / Kidney diseases / Kinases / Life Sciences / Liver / Membrane proteins / Microarrays / Microbial Genetics and Genomics / Nitric oxide / p38 Mitogen-Activated Protein Kinases - metabolism / Plant Genetics and Genomics / Protein binding / Proteins / Proteomics / Receptors, G-Protein-Coupled - genetics / Receptors, G-Protein-Coupled - metabolism / Research Article / Signal Transduction / Software / Statistical analysis