Identification of Senescent Cells in the Bone Microenvironment

Joshua N. Farr, Daniel G. Fraser, Haitao Wang, Katharina Jaehn, Mikolaj B. Ogrodnik, Megan M. Weivoda, Matthew T. Drake, Tamara Tchkonia, Nathan K. LeBrasseur, James L. Kirkland, Lynda Bonewald, Robert J. Pignolo, David G. Monroe, Sundeep Khosla

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

Cellular senescence is a fundamental mechanism by which cells remain metabolically active yet cease dividing and undergo distinct phenotypic alterations, including upregulation of p16Ink4a, profound secretome changes, telomere shortening, and decondensation of pericentromeric satellite DNA. Because senescent cells accumulate in multiple tissues with aging, these cells and the dysfunctional factors they secrete, termed the senescence-associated secretory phenotype (SASP), are increasingly recognized as promising therapeutic targets to prevent age-related degenerative pathologies, including osteoporosis. However, the cell type(s) within the bone microenvironment that undergoes senescence with aging in vivo has remained poorly understood, largely because previous studies have focused on senescence in cultured cells. Thus in young (age 6 months) and old (age 24 months) mice, we measured senescence and SASP markers in vivo in highly enriched cell populations, all rapidly isolated from bone/marrow without in vitro culture. In both females and males, p16Ink4a expression by real-time quantitative polymerase chain reaction (rt-qPCR) was significantly higher with aging in B cells, T cells, myeloid cells, osteoblast progenitors, osteoblasts, and osteocytes. Further, in vivo quantification of senescence-associated distension of satellites (SADS), ie, large-scale unraveling of pericentromeric satellite DNA, revealed significantly more senescent osteocytes in old compared with young bone cortices (11% versus 2%, p < 0.001). In addition, primary osteocytes from old mice had sixfold more (p < 0.001) telomere dysfunction-induced foci (TIFs) than osteocytes from young mice. Corresponding with the age-associated accumulation of senescent osteocytes was significantly higher expression of multiple SASP markers in osteocytes from old versus young mice, several of which also showed dramatic age-associated upregulation in myeloid cells. These data show that with aging, a subset of cells of various lineages within the bone microenvironment become senescent, although senescent myeloid cells and senescent osteocytes predominantly develop the SASP. Given the critical roles of osteocytes in orchestrating bone remodeling, our findings suggest that senescent osteocytes and their SASP may contribute to age-related bone loss.

Original languageEnglish (US)
Pages (from-to)1920-1929
Number of pages10
JournalJournal of Bone and Mineral Research
Volume31
Issue number11
DOIs
StatePublished - Nov 1 2016
Externally publishedYes

Fingerprint

Osteocytes
Bone and Bones
Phenotype
Satellite DNA
Cell Aging
Myeloid Cells
Osteoblasts
Osteoporosis
Up-Regulation
Telomere Shortening
Myeloid Progenitor Cells
Bone Remodeling
Telomere
Cell Lineage
Real-Time Polymerase Chain Reaction
Cultured Cells
B-Lymphocytes
Bone Marrow
Pathology
T-Lymphocytes

Keywords

  • AGING
  • ANIMAL MODELS
  • CELL/TISSUE SIGNALING
  • OSTEOCYTES

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Orthopedics and Sports Medicine

Cite this

Farr, J. N., Fraser, D. G., Wang, H., Jaehn, K., Ogrodnik, M. B., Weivoda, M. M., ... Khosla, S. (2016). Identification of Senescent Cells in the Bone Microenvironment. Journal of Bone and Mineral Research, 31(11), 1920-1929. https://doi.org/10.1002/jbmr.2892

Identification of Senescent Cells in the Bone Microenvironment. / Farr, Joshua N.; Fraser, Daniel G.; Wang, Haitao; Jaehn, Katharina; Ogrodnik, Mikolaj B.; Weivoda, Megan M.; Drake, Matthew T.; Tchkonia, Tamara; LeBrasseur, Nathan K.; Kirkland, James L.; Bonewald, Lynda; Pignolo, Robert J.; Monroe, David G.; Khosla, Sundeep.

In: Journal of Bone and Mineral Research, Vol. 31, No. 11, 01.11.2016, p. 1920-1929.

Research output: Contribution to journalArticle

Farr, JN, Fraser, DG, Wang, H, Jaehn, K, Ogrodnik, MB, Weivoda, MM, Drake, MT, Tchkonia, T, LeBrasseur, NK, Kirkland, JL, Bonewald, L, Pignolo, RJ, Monroe, DG & Khosla, S 2016, 'Identification of Senescent Cells in the Bone Microenvironment', Journal of Bone and Mineral Research, vol. 31, no. 11, pp. 1920-1929. https://doi.org/10.1002/jbmr.2892
Farr JN, Fraser DG, Wang H, Jaehn K, Ogrodnik MB, Weivoda MM et al. Identification of Senescent Cells in the Bone Microenvironment. Journal of Bone and Mineral Research. 2016 Nov 1;31(11):1920-1929. https://doi.org/10.1002/jbmr.2892
Farr, Joshua N. ; Fraser, Daniel G. ; Wang, Haitao ; Jaehn, Katharina ; Ogrodnik, Mikolaj B. ; Weivoda, Megan M. ; Drake, Matthew T. ; Tchkonia, Tamara ; LeBrasseur, Nathan K. ; Kirkland, James L. ; Bonewald, Lynda ; Pignolo, Robert J. ; Monroe, David G. ; Khosla, Sundeep. / Identification of Senescent Cells in the Bone Microenvironment. In: Journal of Bone and Mineral Research. 2016 ; Vol. 31, No. 11. pp. 1920-1929.
@article{b829873adcf14bacb2d2406c5738565b,
title = "Identification of Senescent Cells in the Bone Microenvironment",
abstract = "Cellular senescence is a fundamental mechanism by which cells remain metabolically active yet cease dividing and undergo distinct phenotypic alterations, including upregulation of p16Ink4a, profound secretome changes, telomere shortening, and decondensation of pericentromeric satellite DNA. Because senescent cells accumulate in multiple tissues with aging, these cells and the dysfunctional factors they secrete, termed the senescence-associated secretory phenotype (SASP), are increasingly recognized as promising therapeutic targets to prevent age-related degenerative pathologies, including osteoporosis. However, the cell type(s) within the bone microenvironment that undergoes senescence with aging in vivo has remained poorly understood, largely because previous studies have focused on senescence in cultured cells. Thus in young (age 6 months) and old (age 24 months) mice, we measured senescence and SASP markers in vivo in highly enriched cell populations, all rapidly isolated from bone/marrow without in vitro culture. In both females and males, p16Ink4a expression by real-time quantitative polymerase chain reaction (rt-qPCR) was significantly higher with aging in B cells, T cells, myeloid cells, osteoblast progenitors, osteoblasts, and osteocytes. Further, in vivo quantification of senescence-associated distension of satellites (SADS), ie, large-scale unraveling of pericentromeric satellite DNA, revealed significantly more senescent osteocytes in old compared with young bone cortices (11{\%} versus 2{\%}, p < 0.001). In addition, primary osteocytes from old mice had sixfold more (p < 0.001) telomere dysfunction-induced foci (TIFs) than osteocytes from young mice. Corresponding with the age-associated accumulation of senescent osteocytes was significantly higher expression of multiple SASP markers in osteocytes from old versus young mice, several of which also showed dramatic age-associated upregulation in myeloid cells. These data show that with aging, a subset of cells of various lineages within the bone microenvironment become senescent, although senescent myeloid cells and senescent osteocytes predominantly develop the SASP. Given the critical roles of osteocytes in orchestrating bone remodeling, our findings suggest that senescent osteocytes and their SASP may contribute to age-related bone loss.",
keywords = "AGING, ANIMAL MODELS, CELL/TISSUE SIGNALING, OSTEOCYTES",
author = "Farr, {Joshua N.} and Fraser, {Daniel G.} and Haitao Wang and Katharina Jaehn and Ogrodnik, {Mikolaj B.} and Weivoda, {Megan M.} and Drake, {Matthew T.} and Tamara Tchkonia and LeBrasseur, {Nathan K.} and Kirkland, {James L.} and Lynda Bonewald and Pignolo, {Robert J.} and Monroe, {David G.} and Sundeep Khosla",
year = "2016",
month = "11",
day = "1",
doi = "10.1002/jbmr.2892",
language = "English (US)",
volume = "31",
pages = "1920--1929",
journal = "Journal of Bone and Mineral Research",
issn = "0884-0431",
publisher = "Wiley-Blackwell",
number = "11",

}

TY - JOUR

T1 - Identification of Senescent Cells in the Bone Microenvironment

AU - Farr, Joshua N.

AU - Fraser, Daniel G.

AU - Wang, Haitao

AU - Jaehn, Katharina

AU - Ogrodnik, Mikolaj B.

AU - Weivoda, Megan M.

AU - Drake, Matthew T.

AU - Tchkonia, Tamara

AU - LeBrasseur, Nathan K.

AU - Kirkland, James L.

AU - Bonewald, Lynda

AU - Pignolo, Robert J.

AU - Monroe, David G.

AU - Khosla, Sundeep

PY - 2016/11/1

Y1 - 2016/11/1

N2 - Cellular senescence is a fundamental mechanism by which cells remain metabolically active yet cease dividing and undergo distinct phenotypic alterations, including upregulation of p16Ink4a, profound secretome changes, telomere shortening, and decondensation of pericentromeric satellite DNA. Because senescent cells accumulate in multiple tissues with aging, these cells and the dysfunctional factors they secrete, termed the senescence-associated secretory phenotype (SASP), are increasingly recognized as promising therapeutic targets to prevent age-related degenerative pathologies, including osteoporosis. However, the cell type(s) within the bone microenvironment that undergoes senescence with aging in vivo has remained poorly understood, largely because previous studies have focused on senescence in cultured cells. Thus in young (age 6 months) and old (age 24 months) mice, we measured senescence and SASP markers in vivo in highly enriched cell populations, all rapidly isolated from bone/marrow without in vitro culture. In both females and males, p16Ink4a expression by real-time quantitative polymerase chain reaction (rt-qPCR) was significantly higher with aging in B cells, T cells, myeloid cells, osteoblast progenitors, osteoblasts, and osteocytes. Further, in vivo quantification of senescence-associated distension of satellites (SADS), ie, large-scale unraveling of pericentromeric satellite DNA, revealed significantly more senescent osteocytes in old compared with young bone cortices (11% versus 2%, p < 0.001). In addition, primary osteocytes from old mice had sixfold more (p < 0.001) telomere dysfunction-induced foci (TIFs) than osteocytes from young mice. Corresponding with the age-associated accumulation of senescent osteocytes was significantly higher expression of multiple SASP markers in osteocytes from old versus young mice, several of which also showed dramatic age-associated upregulation in myeloid cells. These data show that with aging, a subset of cells of various lineages within the bone microenvironment become senescent, although senescent myeloid cells and senescent osteocytes predominantly develop the SASP. Given the critical roles of osteocytes in orchestrating bone remodeling, our findings suggest that senescent osteocytes and their SASP may contribute to age-related bone loss.

AB - Cellular senescence is a fundamental mechanism by which cells remain metabolically active yet cease dividing and undergo distinct phenotypic alterations, including upregulation of p16Ink4a, profound secretome changes, telomere shortening, and decondensation of pericentromeric satellite DNA. Because senescent cells accumulate in multiple tissues with aging, these cells and the dysfunctional factors they secrete, termed the senescence-associated secretory phenotype (SASP), are increasingly recognized as promising therapeutic targets to prevent age-related degenerative pathologies, including osteoporosis. However, the cell type(s) within the bone microenvironment that undergoes senescence with aging in vivo has remained poorly understood, largely because previous studies have focused on senescence in cultured cells. Thus in young (age 6 months) and old (age 24 months) mice, we measured senescence and SASP markers in vivo in highly enriched cell populations, all rapidly isolated from bone/marrow without in vitro culture. In both females and males, p16Ink4a expression by real-time quantitative polymerase chain reaction (rt-qPCR) was significantly higher with aging in B cells, T cells, myeloid cells, osteoblast progenitors, osteoblasts, and osteocytes. Further, in vivo quantification of senescence-associated distension of satellites (SADS), ie, large-scale unraveling of pericentromeric satellite DNA, revealed significantly more senescent osteocytes in old compared with young bone cortices (11% versus 2%, p < 0.001). In addition, primary osteocytes from old mice had sixfold more (p < 0.001) telomere dysfunction-induced foci (TIFs) than osteocytes from young mice. Corresponding with the age-associated accumulation of senescent osteocytes was significantly higher expression of multiple SASP markers in osteocytes from old versus young mice, several of which also showed dramatic age-associated upregulation in myeloid cells. These data show that with aging, a subset of cells of various lineages within the bone microenvironment become senescent, although senescent myeloid cells and senescent osteocytes predominantly develop the SASP. Given the critical roles of osteocytes in orchestrating bone remodeling, our findings suggest that senescent osteocytes and their SASP may contribute to age-related bone loss.

KW - AGING

KW - ANIMAL MODELS

KW - CELL/TISSUE SIGNALING

KW - OSTEOCYTES

UR - http://www.scopus.com/inward/record.url?scp=84997216961&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84997216961&partnerID=8YFLogxK

U2 - 10.1002/jbmr.2892

DO - 10.1002/jbmr.2892

M3 - Article

VL - 31

SP - 1920

EP - 1929

JO - Journal of Bone and Mineral Research

JF - Journal of Bone and Mineral Research

SN - 0884-0431

IS - 11

ER -