Transcriptomic characterization of signaling pathways associated with osteoblastic differentiation of MC-3T3E1 cells

• Published in: PloS one Vol. 14; no. 1; p. e0204197
• Main Authors: Luttrell, Louis M, Dar, Moahad S, Gesty-Palmer, Diane, El-Shewy, Hesham M, Robinson, Katherine M, Haycraft, Courtney J, Barth, Jeremy L
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 04.01.2019; Public Library of Science (PLoS)
• Subjects: 3T3 Cells; Animals; Apoptosis; Autocrine Communication - genetics; Autocrine signalling; Biocompatibility; Biology; Biology and Life Sciences; Biomedical materials; Bone Density - physiology; Bone mineral density; Bone remodeling; Bone Remodeling - genetics; Cell culture; Cell Differentiation - genetics; Computer and Information Sciences; Coordination compounds; Cytokines; Datasets; Datasets as Topic; Deoxyribonucleic acid; Differentiation; DNA; DNA binding proteins; DNA chips; DNA microarrays; Endocrinology; Extracellular matrix; Extracellular Matrix - physiology; Gene expression; Gene Expression Profiling; Gene Regulatory Networks - physiology; Growth factors; Hormones; Kinases; Maturation; Mechanical properties; Medicine; Medicine and Health Sciences; Mesenchyme; Metabolism; Mice; Mineralization; Network analysis; Oligonucleotide Array Sequence Analysis; Osteoblastogenesis; Osteoblasts; Osteoblasts - physiology; Osteoclastogenesis; Osteoclasts; Osteogenesis - genetics; Paracrine Communication - genetics; Paracrine signalling; Pluripotency; Proteins; Research and Analysis Methods; Signal transduction; Signal Transduction - genetics; Stem cells; TRANCE protein; Transcriptome - physiology; Tumor necrosis factor-TNF; Tumor necrosis factor-α; Wnt protein; β-Catenin; North Carolina; Mississippi; United States > US; South Carolina
• DOI: 10.1371/journal.pone.0204197
• ISSN: 1932-6203

Bone remodeling involves the coordinated actions of osteoclasts, which resorb the calcified bony matrix, and osteoblasts, which refill erosion pits created by osteoclasts to restore skeletal integrity and adapt to changes in mechanical load. Osteoblasts are derived from pluripotent mesenchymal stem cell precursors, which undergo differentiation under the influence of a host of local and environmental cues. To characterize the autocrine/paracrine signaling networks associated with osteoblast maturation and function, we performed gene network analysis using complementary "agnostic" DNA microarray and "targeted" NanoString nCounter datasets derived from murine MC3T3-E1 cells induced to undergo synchronized osteoblastic differentiation in vitro. Pairwise datasets representing changes in gene expression associated with growth arrest (day 2 to 5 in culture), differentiation (day 5 to 10 in culture), and osteoblast maturation (day 10 to 28 in culture) were analyzed using Ingenuity Systems Pathways Analysis to generate predictions about signaling pathway activity based on the temporal sequence of changes in target gene expression. Our data indicate that some pathways involved in osteoblast differentiation, e.g. Wnt/β-catenin signaling, are most active early in the process, while others, e.g. TGFβ/BMP, cytokine/JAK-STAT and TNFα/RANKL signaling, increase in activity as differentiation progresses. Collectively, these pathways contribute to the sequential expression of genes involved in the synthesis and mineralization of extracellular matrix. These results provide insight into the temporal coordination and complex interplay between signaling networks controlling gene expression during osteoblast differentiation. A more complete understanding of these processes may aid the discovery of novel methods to promote osteoblast development for the treatment of conditions characterized by low bone mineral density.