Subcellular localization of IRS-1 in IGF-I-mediated chondrogenic proliferation, differentiation and hypertrophy of bone marrow mesenchymal stem cells

Lara Longobardi, Froilán Granero-Moltó, Lynda O'Rear, Timothy J. Myers, Tieshi Li, Philip J. Kregor, Anna Spagnoli

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Bone marrow derived mesenchymal stem cells (BM-MSC) can differentiate into chondrocytes. Understanding the mechanisms and growth factors that control the MSC stemness is critical to fully implement their therapeutic use in cartilage diseases. The activated type 1 insulin-like growth factor receptor (IGF-IR), interacting with the insulin receptor substrate-1 (IRS-1), can induce cancer cell proliferation and transformation. In cancer or transformed cells, IRS-1 has been shown to localize in the cytoplasm where it activates the canonical Akt pathway, as well as in the nucleus where it binds to nuclear proteins. We have previously demonstrated that IGF-I has distinct time-dependent effect on primary BM-MSC chondrogenic pellets: initially (2-day culture), IGF-I induces proliferation; subsequently, IGF-I promotes chondrocytic differentiation (7-day culture). In the present study, by using MSC from the BM of IRS-1-/- mice we show that IRS-1 mediates almost 50% of the IGF-I mitogenic response and the MAPK-MEK/ERK signalling accounts for the other 50%. After stimulation with IGF-I, we found that in 2-day old human and mouse derived BM-MSC pellets, IRS-1 (total and phosphorylated) is nuclearly localized and that proliferation prevails over differentiation. The IGF-I mitogenic effect is Akt-independent. In 7-day MSC pellets, IGF-I stimulates the chondrogenic differentiation of MSC into chondrocytes, pre-hypertrophic and hypertrophic chondrocytes and IRS-1 accumulates in the cytoplasm. IGF-I-dependent differentiation is exclusively Akt-dependent. Our data indicate that in the physiologically relevant model of primary cultured MSC, IGF-I induces a temporally regulated nuclear or cytoplasmic localization of IRS-1 that correlate with the transition from proliferation to chondrogenic differentiation.

Original languageEnglish (US)
Pages (from-to)309-320
Number of pages12
JournalGrowth Factors
Volume27
Issue number5
DOIs
StatePublished - Sep 30 2009

Fingerprint

Insulin Receptor Substrate Proteins
Stem cells
Mesenchymal Stromal Cells
Insulin-Like Growth Factor I
Hypertrophy
Bone
Bone Marrow
Chondrocytes
Cytoplasm
Cartilage Diseases
IGF Type 1 Receptor
Mitogen-Activated Protein Kinase Kinases
Cell proliferation
Cartilage
Therapeutic Uses
Nuclear Proteins
Cell culture
Neoplasms
Intercellular Signaling Peptides and Proteins
Cell Proliferation

Keywords

  • Cell signalling
  • Chondrocyte differentiation
  • IGF-I
  • IRS-1
  • Mesenchymal stem cells

ASJC Scopus subject areas

  • Endocrinology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Subcellular localization of IRS-1 in IGF-I-mediated chondrogenic proliferation, differentiation and hypertrophy of bone marrow mesenchymal stem cells. / Longobardi, Lara; Granero-Moltó, Froilán; O'Rear, Lynda; Myers, Timothy J.; Li, Tieshi; Kregor, Philip J.; Spagnoli, Anna.

In: Growth Factors, Vol. 27, No. 5, 30.09.2009, p. 309-320.

Research output: Contribution to journalArticle

Longobardi, Lara ; Granero-Moltó, Froilán ; O'Rear, Lynda ; Myers, Timothy J. ; Li, Tieshi ; Kregor, Philip J. ; Spagnoli, Anna. / Subcellular localization of IRS-1 in IGF-I-mediated chondrogenic proliferation, differentiation and hypertrophy of bone marrow mesenchymal stem cells. In: Growth Factors. 2009 ; Vol. 27, No. 5. pp. 309-320.
@article{f0b0f7d43b1c4e89af4dc9742e0df404,
title = "Subcellular localization of IRS-1 in IGF-I-mediated chondrogenic proliferation, differentiation and hypertrophy of bone marrow mesenchymal stem cells",
abstract = "Bone marrow derived mesenchymal stem cells (BM-MSC) can differentiate into chondrocytes. Understanding the mechanisms and growth factors that control the MSC stemness is critical to fully implement their therapeutic use in cartilage diseases. The activated type 1 insulin-like growth factor receptor (IGF-IR), interacting with the insulin receptor substrate-1 (IRS-1), can induce cancer cell proliferation and transformation. In cancer or transformed cells, IRS-1 has been shown to localize in the cytoplasm where it activates the canonical Akt pathway, as well as in the nucleus where it binds to nuclear proteins. We have previously demonstrated that IGF-I has distinct time-dependent effect on primary BM-MSC chondrogenic pellets: initially (2-day culture), IGF-I induces proliferation; subsequently, IGF-I promotes chondrocytic differentiation (7-day culture). In the present study, by using MSC from the BM of IRS-1-/- mice we show that IRS-1 mediates almost 50{\%} of the IGF-I mitogenic response and the MAPK-MEK/ERK signalling accounts for the other 50{\%}. After stimulation with IGF-I, we found that in 2-day old human and mouse derived BM-MSC pellets, IRS-1 (total and phosphorylated) is nuclearly localized and that proliferation prevails over differentiation. The IGF-I mitogenic effect is Akt-independent. In 7-day MSC pellets, IGF-I stimulates the chondrogenic differentiation of MSC into chondrocytes, pre-hypertrophic and hypertrophic chondrocytes and IRS-1 accumulates in the cytoplasm. IGF-I-dependent differentiation is exclusively Akt-dependent. Our data indicate that in the physiologically relevant model of primary cultured MSC, IGF-I induces a temporally regulated nuclear or cytoplasmic localization of IRS-1 that correlate with the transition from proliferation to chondrogenic differentiation.",
keywords = "Cell signalling, Chondrocyte differentiation, IGF-I, IRS-1, Mesenchymal stem cells",
author = "Lara Longobardi and Froil{\'a}n Granero-Molt{\'o} and Lynda O'Rear and Myers, {Timothy J.} and Tieshi Li and Kregor, {Philip J.} and Anna Spagnoli",
year = "2009",
month = "9",
day = "30",
doi = "10.1080/08977190903138874",
language = "English (US)",
volume = "27",
pages = "309--320",
journal = "Growth Factors",
issn = "0897-7194",
publisher = "Informa Healthcare",
number = "5",

}

TY - JOUR

T1 - Subcellular localization of IRS-1 in IGF-I-mediated chondrogenic proliferation, differentiation and hypertrophy of bone marrow mesenchymal stem cells

AU - Longobardi, Lara

AU - Granero-Moltó, Froilán

AU - O'Rear, Lynda

AU - Myers, Timothy J.

AU - Li, Tieshi

AU - Kregor, Philip J.

AU - Spagnoli, Anna

PY - 2009/9/30

Y1 - 2009/9/30

N2 - Bone marrow derived mesenchymal stem cells (BM-MSC) can differentiate into chondrocytes. Understanding the mechanisms and growth factors that control the MSC stemness is critical to fully implement their therapeutic use in cartilage diseases. The activated type 1 insulin-like growth factor receptor (IGF-IR), interacting with the insulin receptor substrate-1 (IRS-1), can induce cancer cell proliferation and transformation. In cancer or transformed cells, IRS-1 has been shown to localize in the cytoplasm where it activates the canonical Akt pathway, as well as in the nucleus where it binds to nuclear proteins. We have previously demonstrated that IGF-I has distinct time-dependent effect on primary BM-MSC chondrogenic pellets: initially (2-day culture), IGF-I induces proliferation; subsequently, IGF-I promotes chondrocytic differentiation (7-day culture). In the present study, by using MSC from the BM of IRS-1-/- mice we show that IRS-1 mediates almost 50% of the IGF-I mitogenic response and the MAPK-MEK/ERK signalling accounts for the other 50%. After stimulation with IGF-I, we found that in 2-day old human and mouse derived BM-MSC pellets, IRS-1 (total and phosphorylated) is nuclearly localized and that proliferation prevails over differentiation. The IGF-I mitogenic effect is Akt-independent. In 7-day MSC pellets, IGF-I stimulates the chondrogenic differentiation of MSC into chondrocytes, pre-hypertrophic and hypertrophic chondrocytes and IRS-1 accumulates in the cytoplasm. IGF-I-dependent differentiation is exclusively Akt-dependent. Our data indicate that in the physiologically relevant model of primary cultured MSC, IGF-I induces a temporally regulated nuclear or cytoplasmic localization of IRS-1 that correlate with the transition from proliferation to chondrogenic differentiation.

AB - Bone marrow derived mesenchymal stem cells (BM-MSC) can differentiate into chondrocytes. Understanding the mechanisms and growth factors that control the MSC stemness is critical to fully implement their therapeutic use in cartilage diseases. The activated type 1 insulin-like growth factor receptor (IGF-IR), interacting with the insulin receptor substrate-1 (IRS-1), can induce cancer cell proliferation and transformation. In cancer or transformed cells, IRS-1 has been shown to localize in the cytoplasm where it activates the canonical Akt pathway, as well as in the nucleus where it binds to nuclear proteins. We have previously demonstrated that IGF-I has distinct time-dependent effect on primary BM-MSC chondrogenic pellets: initially (2-day culture), IGF-I induces proliferation; subsequently, IGF-I promotes chondrocytic differentiation (7-day culture). In the present study, by using MSC from the BM of IRS-1-/- mice we show that IRS-1 mediates almost 50% of the IGF-I mitogenic response and the MAPK-MEK/ERK signalling accounts for the other 50%. After stimulation with IGF-I, we found that in 2-day old human and mouse derived BM-MSC pellets, IRS-1 (total and phosphorylated) is nuclearly localized and that proliferation prevails over differentiation. The IGF-I mitogenic effect is Akt-independent. In 7-day MSC pellets, IGF-I stimulates the chondrogenic differentiation of MSC into chondrocytes, pre-hypertrophic and hypertrophic chondrocytes and IRS-1 accumulates in the cytoplasm. IGF-I-dependent differentiation is exclusively Akt-dependent. Our data indicate that in the physiologically relevant model of primary cultured MSC, IGF-I induces a temporally regulated nuclear or cytoplasmic localization of IRS-1 that correlate with the transition from proliferation to chondrogenic differentiation.

KW - Cell signalling

KW - Chondrocyte differentiation

KW - IGF-I

KW - IRS-1

KW - Mesenchymal stem cells

UR - http://www.scopus.com/inward/record.url?scp=70349411641&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=70349411641&partnerID=8YFLogxK

U2 - 10.1080/08977190903138874

DO - 10.1080/08977190903138874

M3 - Article

C2 - 19639489

AN - SCOPUS:70349411641

VL - 27

SP - 309

EP - 320

JO - Growth Factors

JF - Growth Factors

SN - 0897-7194

IS - 5

ER -