TY - JOUR
T1 - RNA-sequencing gene expression profiling of orbital adipose-derived stem cell population implicate HOX genes and WNT signaling dysregulation in the pathogenesis of thyroid-associated orbitopathy
AU - Tao, Wensi
AU - Ayala-Haedo, Juan A.
AU - Field, Matthew G.
AU - Pelaez, Daniel
AU - Wester, Sara T.
N1 - Funding Information:
Supported by grants from the Dr. Nasser Al-Rashid Orbital Research Fund (Miami, FL, USA). Imaging and RGC functional experiments were supported by the National Institutes of Health Center Core Grant P30EY014801 (Bethesda, MD, USA) and Research to Prevent Blindness Unrestricted Grant (New York, NY, USA).
Funding Information:
Supported by grants from the Dr. Nasser Al-Rashid Orbital Research Fund (Miami, FL, USA). Imaging and RGC functional experiments were supported by the National Institutes of Health Center Core Grant P30EY014801 (Bethesda, MD, USA) and Research to Prevent Blindness Unrestricted Grant (New York, NY, USA). Disclosure: W. Tao, None; J.A. Ayala-Haedo, None; M.G. Field, None; D. Pelaez, None; S.T. Wester, None
Publisher Copyright:
© 2017 The Authors.
PY - 2017/12
Y1 - 2017/12
N2 - PURPOSE. The purpose of this study was to characterize the intrinsic cellular properties of orbital adipose-derived stem cells (OASC) from patients with thyroid-associated orbitopathy (TAO) and healthy controls. METHODS. Orbital adipose tissue was collected from a total of nine patients: four controls and five patients with TAO. Isolated OASC were characterized with mesenchymal stem cell-specific markers. Orbital adipose-derived stem cells were differentiated into three lineages: chondrocytes, osteocytes, and adipocytes. Reverse transcription PCR of genes involved in the adipogenesis, chondrogenesis, and osteogenesis pathways were selected to assay the differentiation capacities. RNA sequencing analysis (RNA-seq) was performed and results were compared to assess for differences in gene expression between TAO and controls. Selected top-ranked results were confirmed by RT-PCR. RESULTS. Orbital adipose-derived stem cells isolated from orbital fat expressed high levels of mesenchymal stem cell markers, but low levels of the pluripotent stem cell markers. Orbital adipose-derived stem cells isolated from TAO patients exhibited an increase in adipogenesis, and a decrease in chondrogenesis and osteogenesis. RNA-seq disclosed 54 differentially expressed genes. In TAO OASC, expression of early neural crest progenitor marker (WNT signaling, ZIC genes and MSX2) was lost. Meanwhile, ectopic expression of HOXB2 and HOXB3 was found in the OASC from TAO. CONCLUSION. Our results suggest that there are intrinsic genetic and cellular differences in the OASC populations derived from TAO patients. The upregulation in adipogenesis in OASC of TAO may be is consistent with the clinical phenotype. Downregulation of early neural crest markers and ectopic expression of HOXB2 and HOXB3 in TAO OASC demonstrate dysregulation of developmental and tissue patterning pathways.
AB - PURPOSE. The purpose of this study was to characterize the intrinsic cellular properties of orbital adipose-derived stem cells (OASC) from patients with thyroid-associated orbitopathy (TAO) and healthy controls. METHODS. Orbital adipose tissue was collected from a total of nine patients: four controls and five patients with TAO. Isolated OASC were characterized with mesenchymal stem cell-specific markers. Orbital adipose-derived stem cells were differentiated into three lineages: chondrocytes, osteocytes, and adipocytes. Reverse transcription PCR of genes involved in the adipogenesis, chondrogenesis, and osteogenesis pathways were selected to assay the differentiation capacities. RNA sequencing analysis (RNA-seq) was performed and results were compared to assess for differences in gene expression between TAO and controls. Selected top-ranked results were confirmed by RT-PCR. RESULTS. Orbital adipose-derived stem cells isolated from orbital fat expressed high levels of mesenchymal stem cell markers, but low levels of the pluripotent stem cell markers. Orbital adipose-derived stem cells isolated from TAO patients exhibited an increase in adipogenesis, and a decrease in chondrogenesis and osteogenesis. RNA-seq disclosed 54 differentially expressed genes. In TAO OASC, expression of early neural crest progenitor marker (WNT signaling, ZIC genes and MSX2) was lost. Meanwhile, ectopic expression of HOXB2 and HOXB3 was found in the OASC from TAO. CONCLUSION. Our results suggest that there are intrinsic genetic and cellular differences in the OASC populations derived from TAO patients. The upregulation in adipogenesis in OASC of TAO may be is consistent with the clinical phenotype. Downregulation of early neural crest markers and ectopic expression of HOXB2 and HOXB3 in TAO OASC demonstrate dysregulation of developmental and tissue patterning pathways.
KW - Orbital adipose-derived stem cell
KW - RNA-seq
KW - Thyroid associated orbitopathy
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UR - http://www.scopus.com/inward/citedby.url?scp=85037656278&partnerID=8YFLogxK
U2 - 10.1167/iovs.17-22237
DO - 10.1167/iovs.17-22237
M3 - Article
C2 - 29214313
AN - SCOPUS:85037656278
VL - 58
SP - 6146
EP - 6158
JO - Investigative Ophthalmology and Visual Science
JF - Investigative Ophthalmology and Visual Science
SN - 0146-0404
IS - 14
ER -