Quantitative temporal proteomic analysis of human embryonic stem cell differentiation into oligodendrocyte progenitor cells

Abstract

Oligodendrocytes (OLs) are glial cells of the central nervous system which produce myelin. Cultured OLs provide immense therapeutic opportunities for treating a variety of neurological conditions. One of the most promising sources for such therapies is human embryonic stem cells (ESCs), as well as providing a model to study human oligodendrocyte development. For these purposes, an investigation of proteome level changes is critical for understanding the process of OL differentiation. In this report, an iTRAQ-based quantitative proteomic approach was used to study multiple steps during oligodendrocyte differentiation including neural precursors (NPCs), glial precursors (GPCs), and oligodendrocyte progenitors (OPCs) compared to undifferentiated embryonic stem cells. Using a 1% false discovery rate cutoff, ~3,145 proteins were quantitated and several demonstrated progressive stage-specific expression. Proteins such as TF, NCAM1, APOE, and WNT5A showed increased expression from the NPC to OPC stage. Several proteins that have demonstrated evidence or been suspected in OL maturation were also found upregulated in OPCs including FABP4, THBS1, BMP1, CRYAB, TF, TNC, COL3A1, TGFBI and EPB41L3. Thus, by providing the first extensive proteomic profiling of human embryonic stem cell differentiation into oligodendrocyte progenitor cells, this study provides many novel proteins that are potentially involved in OL development.

The PI3K/Akt/mTOR Pathway Is Implicated in the Premature Senescence of Primary Human Endothelial Cells Exposed to Chronic Radiation

“…Data processing for protein identification and quantification of ICPL- labeled proteins was performed using Proteome Discoverer version 1.3.0.339 (Thermo Scientific) as described before [23], [27]–[29]. Briefly, Proteome Discoverer software performed automated statistical analysis of the results and used unique peptides to calculate accurate relative protein quantification.…”

Members of the high mobility group B protein family are dynamically expressed in embryonic neural stem cells

“…Following up on the role of HMGA2 in regulating p16 Ink4a expression and NSC proliferation, additional studies have reported that the HMGA family of proteins maintain NSC/NPC chromatin in the open state during early development [19]. Moreover, HMGA proteins were identified by iTRAQ proteomic analysis as being expressed during oligodendrocyte precursor cell (OPC) differentiation [20], and transcriptional profiling studies have found that NSCs express HMGB1, HMGB2 and HMGB3 mRNA [21]. A recent study in zebrafish reported that HMGB1 is critical for early brain development, since morpholino-knockdown of HMGB1 expression created forebrain defects and ablation of the catecholamine network [22].…”

Human Embryonic Stem Cell-Derived Oligodendrocytes: Protocols and Perspectives

“…For instance, Letzen et al, showed miRNA profiles during different stages of oligodendrocyte differentiation from hESCs, which possibly presents key markers of oligodendrocyte maturation in vivo [109]. Further, Chaerkady et al, demonstrated several novel proteins, such as fatty acid-binding protein and thrombospondin-1, which are probably involved in oligodendrocyte differentiation and maturation, by presenting an extensive proteomic profiling of oligodendrocytes derived from hESCs [110]. It is through these studies that a comprehensive understanding of molecular and cellular regulators of oligodendrocyte development will be achieved, providing investigators with the most powerful selection criteria for transplantation studies.…”