Combined Characterization of microRNA and mRNA Profiles Delineates Early Differentiation Pathways of CD133+ and CD34+ Hematopoietic Stem and Progenitor Cells

Abstract

MicroRNAs (miRNAs) have been shown to play an important role in hematopoiesis. To elucidate the role of miRNAs in the early steps of hematopoiesis, we directly compared donor-matched CD133+ cells with the more differentiated CD34+CD133− and CD34−CD133− cells from bone marrow on the miRNA and mRNA level. Using quantitative whole genome miRNA microarray and sequencing-based profiling, we found that between 109 (CD133+) and 216 (CD34−CD133−) miRNAs were expressed. Quantification revealed that the 25 highest expressed miRNAs accounted for 73% of the total miRNA pool. miR-142-3p was the highest expressed miRNA with up to 2,000 copies per cell in CD34+CD133− cells. Eighteen miRNAs were significantly differentially expressed between CD133+ and CD34+CD133− cells. We analyzed their biological role by examining the coexpression of miRNAs and its bioinformatically predicted mRNA targets and luciferase-based reporter assays. We provide the first evidence for a direct regulation of CD133 by miR-142-3p as well as tropomyosin 1 and frizzled homolog 5 by miR-29a. Overexpression of miRNAs in CD133+ cells demonstrated that miR-142-3p has a negative influence on the overall colony-forming ability. In conclusion, the miRNAs expressed differentially between the CD133+ and CD34+CD133− cells are involved in inhibition of differentiation, prevention of apoptosis, and cytoskeletal remodeling. These results are highly relevant for stem cell-based therapies with CD133+ cells and delineate for the first time how the stem cell character of CD133+ cells is defined by the expression of specific miRNAs. Stem Cells 2011;29:847–857

miR-142-3p regulates the formation and differentiation of hematopoietic stem cells in vertebrates

“…Interestingly, a very recent report showed that in Xenopus, miR-142-3p is required for HSC lineage specification [37], which is consistent with the results for zebrafish and mouse embryos reported in this study. In contrast, an inverse correlation between miR-142-3p expression and HSC development has been suggested recently [20]. Bissels et al [20] reported that overexpression of miR-142-3p at a dose of 50 nM in human CD133 + cells had a negative effect on their overall colony-forming ability.…”

“…To date, very few miRNAs have been shown to play important roles in HSC development in vertebrates in vivo. miR-142-3p is specifically expressed in HSCs (AGM and CHT) and T cells (thymus) in zebrafish, mouse and other species [15][16][17][18][19][20][21]. By knockdown of miR-142a-3p using 2 specific MOs, the numbers of HSCs and T cells were decreased.…”

“…In addition, miR-144 and miR-451, as direct targets of the critical hematopoietic master regulator GATA1, are required for erythropoiesis in zebrafish and mouse [11][12][13]. miR-142-3p is an evolutionally conserved miRNA of vertebrates, which is expressed in many different hematopoietic cells [14][15][16][17][18][19][20][21]. Chen et al [15] first reported that miR-142 is expressed in embryonic and adult hematopoietic tissues, including the fetal liver, bone marrow, spleen and thymus, in mice, indicating its role in both embryonic and adult hematopoiesis.…”

Demethylation of microRNA-142 induced by demethylation agents plays a suppressive role in osteosarcoma cells

“…It was also observed that increased miR-142 expression by Aza or PBA repressed cell migration. The overexpression of miR-142 has been shown to inhibit the proliferation and colony-forming ability of primitive hematopoietic cells (31). Some of these functions appear to depend on the cellular type and context, since it was reported that miR-142 acts as an anti-migratory factor in hepatocellular carcinoma cells (32) and has also been reported to be deregulated in mesenchymal tumors (33).…”

Bir Gen İfade Düzenleyicisi miRNA

“…mRNA'ların negatif post-transkripsiyonel düzenleyicileri olan 18-25nt uzunluğundaki miRNA'ların her biri mRNA'nın yüzlercesine bağlanıp gen ekspresyonunu düzenlerken ya da bir mRNA yüzlerce miRNA tarafından ekspresyon seviyeleri düzenlenmektedir 10, 65 . Büyüme, gelişme, farklılaşma ve apoptozis gibi birçok biyolojik süreçte işlevi olan miRNA'ların çeşitli hastalıkların tanı, prognoz ve tedavisinde kullanılmaya başlanmıştır 66,67 .…”