NR2F1 mediated down-regulation of osteoblast differentiation was rescued by bone morphogenetic protein-2 (BMP-2) in human MSC

Muthurangan Manikandan, Sarah Abuelreich, Mona Elsafadi, Hussain Alsalman, Hassan Almalak, Abdulaziz Siyal, Jamil Amjad Hashmi, Abdullah Aldahmash, Moustapha Kassem, Musaad Alfayez, Amer Mahmood*

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Resumé

Endochondral ossification is the process by which long bones are formed; the process of long bone formation is regulated by numerous factors such as transcription factors, cytokines, and extracellular matrix molecules. Human hormone Nuclear receptors (hHNR) are a family of ligand-regulated transcription factors that are activated by steroid hormones, such as estrogen and progesterone, and various lipid-soluble signals, including retinoic acid, oxysterols, and thyroid hormone. Whole genome microarray data from our previous study revealed that most hHNR's are up-regulated during osteoblast differentiation in hMSCS. NR2F1 was among the highest expressed hHNR during osteogenesis, NR2F1 belongs to the steroid/thyroid hormone nuclear receptor superfamily. NR2F1 is designated as an orphan nuclear receptor because its ligands are unknown. NR2F1 plays a wide range of roles, including cell differentiation, cancer progression, and central and peripheral neurogenesis. Identifying signaling networks involved in osteoblast differentiation is important in orchestrating new therapeutic and clinical applications in bone biology. This study aimed to identify alterations in signaling networks mediated by NR2F1 in osteoblast differentiation. siRNA-mediated down-regulation of NR2F1 leads to impairment in the differentiation of hBMSC-TERT to osteoblast; gene-expression results confirmed the down-regulation of osteoblast markers such as RUNX2, ALPL, OSC, and BSP. Global whole gene expression analysis revealed that most down-regulated genes were associated with osteoblast differentiation (DDIT3, BMP2). Pathway analysis revealed prominent signaling pathways that were down-regulated, including the TGFβ pathway and MAPK pathway. Functional studies on NR2F1 transfected cells, during osteoblast differentiation in combination with TGFβ1 and BMP-2, showed that TGFβ1 does not recover osteoblast differentiation, whereas BMP-2 rescues osteoblast differentiation in NR2F1 siRNA transfected cells. Thus, our results showed that BMP-2 could intervene in NR2F1 down-regulated signaling pathways to recover osteoblast differentiation.

OriginalsprogEngelsk
TidsskriftDifferentiation
Vol/bind104
Sider (fra-til)36-41
ISSN0301-4681
DOI
StatusUdgivet - 1. nov. 2018

Fingeraftryk

Osteoblasts
Down-Regulation
Cytoplasmic and Nuclear Receptors
Osteogenesis
Small Interfering RNA
human BMP2 protein
Ligands
Thyroid Hormone Receptors
Cell Differentiation
Estrogens
Hormones
Lipids

Citer dette

Manikandan, Muthurangan ; Abuelreich, Sarah ; Elsafadi, Mona ; Alsalman, Hussain ; Almalak, Hassan ; Siyal, Abdulaziz ; Hashmi, Jamil Amjad ; Aldahmash, Abdullah ; Kassem, Moustapha ; Alfayez, Musaad ; Mahmood, Amer. / NR2F1 mediated down-regulation of osteoblast differentiation was rescued by bone morphogenetic protein-2 (BMP-2) in human MSC. I: Differentiation. 2018 ; Bind 104. s. 36-41.
@article{dbc9a5f7625f4e8685d7aa1e4d857ab8,
title = "NR2F1 mediated down-regulation of osteoblast differentiation was rescued by bone morphogenetic protein-2 (BMP-2) in human MSC",
abstract = "Endochondral ossification is the process by which long bones are formed; the process of long bone formation is regulated by numerous factors such as transcription factors, cytokines, and extracellular matrix molecules. Human hormone Nuclear receptors (hHNR) are a family of ligand-regulated transcription factors that are activated by steroid hormones, such as estrogen and progesterone, and various lipid-soluble signals, including retinoic acid, oxysterols, and thyroid hormone. Whole genome microarray data from our previous study revealed that most hHNR's are up-regulated during osteoblast differentiation in hMSCS. NR2F1 was among the highest expressed hHNR during osteogenesis, NR2F1 belongs to the steroid/thyroid hormone nuclear receptor superfamily. NR2F1 is designated as an orphan nuclear receptor because its ligands are unknown. NR2F1 plays a wide range of roles, including cell differentiation, cancer progression, and central and peripheral neurogenesis. Identifying signaling networks involved in osteoblast differentiation is important in orchestrating new therapeutic and clinical applications in bone biology. This study aimed to identify alterations in signaling networks mediated by NR2F1 in osteoblast differentiation. siRNA-mediated down-regulation of NR2F1 leads to impairment in the differentiation of hBMSC-TERT to osteoblast; gene-expression results confirmed the down-regulation of osteoblast markers such as RUNX2, ALPL, OSC, and BSP. Global whole gene expression analysis revealed that most down-regulated genes were associated with osteoblast differentiation (DDIT3, BMP2). Pathway analysis revealed prominent signaling pathways that were down-regulated, including the TGFβ pathway and MAPK pathway. Functional studies on NR2F1 transfected cells, during osteoblast differentiation in combination with TGFβ1 and BMP-2, showed that TGFβ1 does not recover osteoblast differentiation, whereas BMP-2 rescues osteoblast differentiation in NR2F1 siRNA transfected cells. Thus, our results showed that BMP-2 could intervene in NR2F1 down-regulated signaling pathways to recover osteoblast differentiation.",
keywords = "BMP2 signaling, DDIT3, Human bone marrow MSCs, NR2F1, Osteogenesis, TGFb1 signaling",
author = "Muthurangan Manikandan and Sarah Abuelreich and Mona Elsafadi and Hussain Alsalman and Hassan Almalak and Abdulaziz Siyal and Hashmi, {Jamil Amjad} and Abdullah Aldahmash and Moustapha Kassem and Musaad Alfayez and Amer Mahmood",
year = "2018",
month = "11",
day = "1",
doi = "10.1016/j.diff.2018.10.003",
language = "English",
volume = "104",
pages = "36--41",
journal = "Differentiation",
issn = "0301-4681",
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Manikandan, M, Abuelreich, S, Elsafadi, M, Alsalman, H, Almalak, H, Siyal, A, Hashmi, JA, Aldahmash, A, Kassem, M, Alfayez, M & Mahmood, A 2018, 'NR2F1 mediated down-regulation of osteoblast differentiation was rescued by bone morphogenetic protein-2 (BMP-2) in human MSC', Differentiation, bind 104, s. 36-41. https://doi.org/10.1016/j.diff.2018.10.003

NR2F1 mediated down-regulation of osteoblast differentiation was rescued by bone morphogenetic protein-2 (BMP-2) in human MSC. / Manikandan, Muthurangan; Abuelreich, Sarah; Elsafadi, Mona; Alsalman, Hussain; Almalak, Hassan; Siyal, Abdulaziz; Hashmi, Jamil Amjad; Aldahmash, Abdullah; Kassem, Moustapha; Alfayez, Musaad; Mahmood, Amer.

I: Differentiation, Bind 104, 01.11.2018, s. 36-41.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - NR2F1 mediated down-regulation of osteoblast differentiation was rescued by bone morphogenetic protein-2 (BMP-2) in human MSC

AU - Manikandan, Muthurangan

AU - Abuelreich, Sarah

AU - Elsafadi, Mona

AU - Alsalman, Hussain

AU - Almalak, Hassan

AU - Siyal, Abdulaziz

AU - Hashmi, Jamil Amjad

AU - Aldahmash, Abdullah

AU - Kassem, Moustapha

AU - Alfayez, Musaad

AU - Mahmood, Amer

PY - 2018/11/1

Y1 - 2018/11/1

N2 - Endochondral ossification is the process by which long bones are formed; the process of long bone formation is regulated by numerous factors such as transcription factors, cytokines, and extracellular matrix molecules. Human hormone Nuclear receptors (hHNR) are a family of ligand-regulated transcription factors that are activated by steroid hormones, such as estrogen and progesterone, and various lipid-soluble signals, including retinoic acid, oxysterols, and thyroid hormone. Whole genome microarray data from our previous study revealed that most hHNR's are up-regulated during osteoblast differentiation in hMSCS. NR2F1 was among the highest expressed hHNR during osteogenesis, NR2F1 belongs to the steroid/thyroid hormone nuclear receptor superfamily. NR2F1 is designated as an orphan nuclear receptor because its ligands are unknown. NR2F1 plays a wide range of roles, including cell differentiation, cancer progression, and central and peripheral neurogenesis. Identifying signaling networks involved in osteoblast differentiation is important in orchestrating new therapeutic and clinical applications in bone biology. This study aimed to identify alterations in signaling networks mediated by NR2F1 in osteoblast differentiation. siRNA-mediated down-regulation of NR2F1 leads to impairment in the differentiation of hBMSC-TERT to osteoblast; gene-expression results confirmed the down-regulation of osteoblast markers such as RUNX2, ALPL, OSC, and BSP. Global whole gene expression analysis revealed that most down-regulated genes were associated with osteoblast differentiation (DDIT3, BMP2). Pathway analysis revealed prominent signaling pathways that were down-regulated, including the TGFβ pathway and MAPK pathway. Functional studies on NR2F1 transfected cells, during osteoblast differentiation in combination with TGFβ1 and BMP-2, showed that TGFβ1 does not recover osteoblast differentiation, whereas BMP-2 rescues osteoblast differentiation in NR2F1 siRNA transfected cells. Thus, our results showed that BMP-2 could intervene in NR2F1 down-regulated signaling pathways to recover osteoblast differentiation.

AB - Endochondral ossification is the process by which long bones are formed; the process of long bone formation is regulated by numerous factors such as transcription factors, cytokines, and extracellular matrix molecules. Human hormone Nuclear receptors (hHNR) are a family of ligand-regulated transcription factors that are activated by steroid hormones, such as estrogen and progesterone, and various lipid-soluble signals, including retinoic acid, oxysterols, and thyroid hormone. Whole genome microarray data from our previous study revealed that most hHNR's are up-regulated during osteoblast differentiation in hMSCS. NR2F1 was among the highest expressed hHNR during osteogenesis, NR2F1 belongs to the steroid/thyroid hormone nuclear receptor superfamily. NR2F1 is designated as an orphan nuclear receptor because its ligands are unknown. NR2F1 plays a wide range of roles, including cell differentiation, cancer progression, and central and peripheral neurogenesis. Identifying signaling networks involved in osteoblast differentiation is important in orchestrating new therapeutic and clinical applications in bone biology. This study aimed to identify alterations in signaling networks mediated by NR2F1 in osteoblast differentiation. siRNA-mediated down-regulation of NR2F1 leads to impairment in the differentiation of hBMSC-TERT to osteoblast; gene-expression results confirmed the down-regulation of osteoblast markers such as RUNX2, ALPL, OSC, and BSP. Global whole gene expression analysis revealed that most down-regulated genes were associated with osteoblast differentiation (DDIT3, BMP2). Pathway analysis revealed prominent signaling pathways that were down-regulated, including the TGFβ pathway and MAPK pathway. Functional studies on NR2F1 transfected cells, during osteoblast differentiation in combination with TGFβ1 and BMP-2, showed that TGFβ1 does not recover osteoblast differentiation, whereas BMP-2 rescues osteoblast differentiation in NR2F1 siRNA transfected cells. Thus, our results showed that BMP-2 could intervene in NR2F1 down-regulated signaling pathways to recover osteoblast differentiation.

KW - BMP2 signaling

KW - DDIT3

KW - Human bone marrow MSCs

KW - NR2F1

KW - Osteogenesis

KW - TGFb1 signaling

U2 - 10.1016/j.diff.2018.10.003

DO - 10.1016/j.diff.2018.10.003

M3 - Journal article

C2 - 30445268

AN - SCOPUS:85056480435

VL - 104

SP - 36

EP - 41

JO - Differentiation

JF - Differentiation

SN - 0301-4681

ER -