TY - JOUR
T1 - Validation of Osteogenic Properties of Cytochalasin D by High-Resolution RNA-Sequencing in Mesenchymal Stem Cells Derived from Bone Marrow and Adipose Tissues
AU - Samsonraj, Rebekah M.
AU - Paradise, Christopher R.
AU - Dudakovic, Amel
AU - Sen, Buer
AU - Nair, Asha A.
AU - Dietz, Allan B.
AU - Deyle, David R.
AU - Cool, Simon M.
AU - Rubin, Janet
AU - Van Wijnen, Andre J.
N1 - Funding Information:
This study received funding support from the U.S. National Institutes of Health (grant nos. R01-AR049069 to A.v.W., R01-AR066616 to J.R., and F32-AR066508 to A.D.) and the Mayo Clinic Center for Regenerative Medicine in addition to generous philanthropic gifts from William and Karen Eby. We appreciate the bioinformatic support provided by the Mayo Clinic Bioinformatics Core and especially thank Jaime Davila, Ying Li, and Jared Evans for sharing their expertise. We also thank all present and former members of our laboratories, particularly Scott Riester, Emily Camilleri, Eric Lewallen, Janet Denbeigh, Roman Thaler, Farzaneh Khani, Catalina Galeano, Martina Gluscevic, and Oksana Pichurin for sharing reagents and ideas as well as stimulating discussions throughout the course of this work.
Publisher Copyright:
© Copyright 2018, Mary Ann Liebert, Inc.
PY - 2018/8/15
Y1 - 2018/8/15
N2 - Differentiation of mesenchymal stromal/stem cells (MSCs) involves a series of molecular signals and gene transcription events required for attaining cell lineage commitment. Modulation of the actin cytoskeleton using cytochalasin D (CytoD) drives osteogenesis at early timepoints in bone marrow-derived MSCs and also initiates a robust osteogenic differentiation program in adipose tissue-derived MSCs. To understand the molecular basis for these pronounced effects on osteogenic differentiation, we investigated global changes in gene expression in CytoD-treated murine and human MSCs by high-resolution RNA-sequencing (RNA-seq) analysis. A three-way bioinformatic comparison between human adipose tissue-derived MSCs (hAMSCs), human bone marrow-derived MSCs (hBMSCs), and mouse bone marrow-derived MSCs (mBMSCs) revealed significant upregulation of genes linked to extracellular matrix organization, cell adhesion and bone metabolism. As anticipated, the activation of these differentiation-related genes is accompanied by a downregulation of nuclear and cell cycle-related genes presumably reflecting cytostatic effects of CytoD. We also identified eight novel CytoD activated genes - VGLL4, ARHGAP24, KLHL24, RCBTB2, BDH2, SCARF2, ACAD10, HEPH - which are commonly upregulated across the two species and tissue sources of our MSC samples. We selected the Hippo pathway-related VGLL4 gene, which encodes the transcriptional co-factor Vestigial-like 4, for further study because this pathway is linked to osteogenesis. VGLL4 small interfering RNA depletion reduces mineralization of hAMSCs during CytoD-induced osteogenic differentiation. Together, our RNA-seq analyses suggest that while the stimulatory effects of CytoD on osteogenesis are pleiotropic and depend on the biological state of the cell type, a small group of genes including VGLL4 may contribute to MSC commitment toward the bone lineage.
AB - Differentiation of mesenchymal stromal/stem cells (MSCs) involves a series of molecular signals and gene transcription events required for attaining cell lineage commitment. Modulation of the actin cytoskeleton using cytochalasin D (CytoD) drives osteogenesis at early timepoints in bone marrow-derived MSCs and also initiates a robust osteogenic differentiation program in adipose tissue-derived MSCs. To understand the molecular basis for these pronounced effects on osteogenic differentiation, we investigated global changes in gene expression in CytoD-treated murine and human MSCs by high-resolution RNA-sequencing (RNA-seq) analysis. A three-way bioinformatic comparison between human adipose tissue-derived MSCs (hAMSCs), human bone marrow-derived MSCs (hBMSCs), and mouse bone marrow-derived MSCs (mBMSCs) revealed significant upregulation of genes linked to extracellular matrix organization, cell adhesion and bone metabolism. As anticipated, the activation of these differentiation-related genes is accompanied by a downregulation of nuclear and cell cycle-related genes presumably reflecting cytostatic effects of CytoD. We also identified eight novel CytoD activated genes - VGLL4, ARHGAP24, KLHL24, RCBTB2, BDH2, SCARF2, ACAD10, HEPH - which are commonly upregulated across the two species and tissue sources of our MSC samples. We selected the Hippo pathway-related VGLL4 gene, which encodes the transcriptional co-factor Vestigial-like 4, for further study because this pathway is linked to osteogenesis. VGLL4 small interfering RNA depletion reduces mineralization of hAMSCs during CytoD-induced osteogenic differentiation. Together, our RNA-seq analyses suggest that while the stimulatory effects of CytoD on osteogenesis are pleiotropic and depend on the biological state of the cell type, a small group of genes including VGLL4 may contribute to MSC commitment toward the bone lineage.
KW - bone
KW - cell signaling
KW - cytochalasin D
KW - mesenchymal stromal cell
KW - osteoblast
KW - osteogenesis
KW - stem cell
UR - http://www.scopus.com/inward/record.url?scp=85051948685&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85051948685&partnerID=8YFLogxK
U2 - 10.1089/scd.2018.0037
DO - 10.1089/scd.2018.0037
M3 - Article
C2 - 29882479
AN - SCOPUS:85051948685
SN - 1547-3287
VL - 27
SP - 1136
EP - 1145
JO - Stem Cells and Development
JF - Stem Cells and Development
IS - 16
ER -