ADAMTS5 Deficiency Protects Mice from Chronic Tobacco Smoking-induced Intervertebral Disc Degeneration

Kevin Ngo, Pedro Pohl, Dong Wang, Adriana S. Leme, Joon Lee, Peter Di, Peter Roughley, Paul D. Robbins, Laura J. Niedernhofer, Gwendolyn Sowa, James D. Kang, Steven S. Shapiro, Nam V. Vo

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Study Design. ADAMTS5-deficient and wild type (WT) mice were chronically exposed to tobacco smoke to investigate effects on intervertebral disc degeneration (IDD). Objective. The aim of this study was to demonstrate a role for ADAMTS5 in mediating tobacco smoking-induced IDD. Summary of Background Data. We previously demonstrated that chronic tobacco smoking causes IDD in mice because, in part, of proteolytic destruction of disc aggrecan. However, it was unknown which matrix proteinase(s) drive these detrimental effects. Methods. Three-month-old WT (C57BL/6) and ADAMTS5-/- mice were chronically exposed to tobacco smoke (four cigarettes/day, 5 day/week for 6 months). ADAMTS-mediated cleavage of disc aggrecan was analyzed by Western blot. Disc total glycosaminoglycan (GAG) content was assessed by dimethyl methylene blue assay and safranin O/fast green histology. Vertebral osteoporosity was measured by microcomputed tomography. Human nucleus pulposus (hNP) cell cultures were also exposed directly to tobacco smoke extract (TSE), a condensate containing the water-soluble compounds inhaled by smokers, to measure ADAMTS5 expression and ADAMTS-mediated cleavage of aggrecan. Activation of nuclear factor (NF)-κB, a family of transcription factors essential for modulating the cellular response to stress, was measured by immunofluorescence assay. Results. Genetic depletion of ADAMTS5 prevented vertebral bone loss, substantially reduced loss of disc GAG content, and completely obviated ADAMTS-mediated proteolysis of disc aggrecan within its interglobular domain (IGD) in mice following exposure to tobacco smoke. hNP cell cultures exposed to TSE also resulted in upregulation of ADAMTS5 protein expression and a concomitant increase in ADAMTS-mediated cleavage within aggrecan IGD. Activation of NF-κB, known to be required for ADAMTS5 gene expression, was observed in both TSE-treated hNP cell cultures and disc tissue of tobacco smoke-exposed mice. Conclusion. The findings demonstrate that ADAMTS5 is the primary aggrecanase mediating smoking-induced disc aggrecanolysis and IDD. Mouse models of chronic tobacco smoking are important and useful for probing the mechanisms of disc aggrecan catabolism and IDD.

Original languageEnglish (US)
Pages (from-to)1521-1528
Number of pages8
JournalSpine
Volume42
Issue number20
DOIs
StatePublished - Oct 15 2017

Fingerprint

Intervertebral Disc Degeneration
Aggrecans
Smoke
Tobacco
Smoking
Cell Culture Techniques
Glycosaminoglycans
X-Ray Microtomography
Methylene Blue
Tobacco Products
Proteolysis
Fluorescent Antibody Technique
Histology
Peptide Hydrolases
Transcription Factors
Up-Regulation
Western Blotting
Gene Expression
Bone and Bones
Water

Keywords

  • ADAMTS5
  • MMPs
  • NF-kB
  • aggrecanases
  • aggrecanolysis
  • cellular senescence
  • genotoxic
  • inflammatory stress
  • intervertebral disc
  • tobacco smoke

Cite this

Ngo, K., Pohl, P., Wang, D., Leme, A. S., Lee, J., Di, P., ... Vo, N. V. (2017). ADAMTS5 Deficiency Protects Mice from Chronic Tobacco Smoking-induced Intervertebral Disc Degeneration. Spine, 42(20), 1521-1528. https://doi.org/10.1097/BRS.0000000000002258

ADAMTS5 Deficiency Protects Mice from Chronic Tobacco Smoking-induced Intervertebral Disc Degeneration. / Ngo, Kevin; Pohl, Pedro; Wang, Dong; Leme, Adriana S.; Lee, Joon; Di, Peter; Roughley, Peter; Robbins, Paul D.; Niedernhofer, Laura J.; Sowa, Gwendolyn; Kang, James D.; Shapiro, Steven S.; Vo, Nam V.

In: Spine, Vol. 42, No. 20, 15.10.2017, p. 1521-1528.

Research output: Contribution to journalArticle

Ngo, K, Pohl, P, Wang, D, Leme, AS, Lee, J, Di, P, Roughley, P, Robbins, PD, Niedernhofer, LJ, Sowa, G, Kang, JD, Shapiro, SS & Vo, NV 2017, 'ADAMTS5 Deficiency Protects Mice from Chronic Tobacco Smoking-induced Intervertebral Disc Degeneration', Spine, vol. 42, no. 20, pp. 1521-1528. https://doi.org/10.1097/BRS.0000000000002258
Ngo, Kevin ; Pohl, Pedro ; Wang, Dong ; Leme, Adriana S. ; Lee, Joon ; Di, Peter ; Roughley, Peter ; Robbins, Paul D. ; Niedernhofer, Laura J. ; Sowa, Gwendolyn ; Kang, James D. ; Shapiro, Steven S. ; Vo, Nam V. / ADAMTS5 Deficiency Protects Mice from Chronic Tobacco Smoking-induced Intervertebral Disc Degeneration. In: Spine. 2017 ; Vol. 42, No. 20. pp. 1521-1528.
@article{16661b4930c047af9745660242669ebf,
title = "ADAMTS5 Deficiency Protects Mice from Chronic Tobacco Smoking-induced Intervertebral Disc Degeneration",
abstract = "Study Design. ADAMTS5-deficient and wild type (WT) mice were chronically exposed to tobacco smoke to investigate effects on intervertebral disc degeneration (IDD). Objective. The aim of this study was to demonstrate a role for ADAMTS5 in mediating tobacco smoking-induced IDD. Summary of Background Data. We previously demonstrated that chronic tobacco smoking causes IDD in mice because, in part, of proteolytic destruction of disc aggrecan. However, it was unknown which matrix proteinase(s) drive these detrimental effects. Methods. Three-month-old WT (C57BL/6) and ADAMTS5-/- mice were chronically exposed to tobacco smoke (four cigarettes/day, 5 day/week for 6 months). ADAMTS-mediated cleavage of disc aggrecan was analyzed by Western blot. Disc total glycosaminoglycan (GAG) content was assessed by dimethyl methylene blue assay and safranin O/fast green histology. Vertebral osteoporosity was measured by microcomputed tomography. Human nucleus pulposus (hNP) cell cultures were also exposed directly to tobacco smoke extract (TSE), a condensate containing the water-soluble compounds inhaled by smokers, to measure ADAMTS5 expression and ADAMTS-mediated cleavage of aggrecan. Activation of nuclear factor (NF)-κB, a family of transcription factors essential for modulating the cellular response to stress, was measured by immunofluorescence assay. Results. Genetic depletion of ADAMTS5 prevented vertebral bone loss, substantially reduced loss of disc GAG content, and completely obviated ADAMTS-mediated proteolysis of disc aggrecan within its interglobular domain (IGD) in mice following exposure to tobacco smoke. hNP cell cultures exposed to TSE also resulted in upregulation of ADAMTS5 protein expression and a concomitant increase in ADAMTS-mediated cleavage within aggrecan IGD. Activation of NF-κB, known to be required for ADAMTS5 gene expression, was observed in both TSE-treated hNP cell cultures and disc tissue of tobacco smoke-exposed mice. Conclusion. The findings demonstrate that ADAMTS5 is the primary aggrecanase mediating smoking-induced disc aggrecanolysis and IDD. Mouse models of chronic tobacco smoking are important and useful for probing the mechanisms of disc aggrecan catabolism and IDD.",
keywords = "ADAMTS5, MMPs, NF-kB, aggrecanases, aggrecanolysis, cellular senescence, genotoxic, inflammatory stress, intervertebral disc, tobacco smoke",
author = "Kevin Ngo and Pedro Pohl and Dong Wang and Leme, {Adriana S.} and Joon Lee and Peter Di and Peter Roughley and Robbins, {Paul D.} and Niedernhofer, {Laura J.} and Gwendolyn Sowa and Kang, {James D.} and Shapiro, {Steven S.} and Vo, {Nam V.}",
year = "2017",
month = "10",
day = "15",
doi = "10.1097/BRS.0000000000002258",
language = "English (US)",
volume = "42",
pages = "1521--1528",
journal = "Spine",
issn = "0362-2436",
publisher = "Lippincott Williams and Wilkins",
number = "20",

}

TY - JOUR

T1 - ADAMTS5 Deficiency Protects Mice from Chronic Tobacco Smoking-induced Intervertebral Disc Degeneration

AU - Ngo, Kevin

AU - Pohl, Pedro

AU - Wang, Dong

AU - Leme, Adriana S.

AU - Lee, Joon

AU - Di, Peter

AU - Roughley, Peter

AU - Robbins, Paul D.

AU - Niedernhofer, Laura J.

AU - Sowa, Gwendolyn

AU - Kang, James D.

AU - Shapiro, Steven S.

AU - Vo, Nam V.

PY - 2017/10/15

Y1 - 2017/10/15

N2 - Study Design. ADAMTS5-deficient and wild type (WT) mice were chronically exposed to tobacco smoke to investigate effects on intervertebral disc degeneration (IDD). Objective. The aim of this study was to demonstrate a role for ADAMTS5 in mediating tobacco smoking-induced IDD. Summary of Background Data. We previously demonstrated that chronic tobacco smoking causes IDD in mice because, in part, of proteolytic destruction of disc aggrecan. However, it was unknown which matrix proteinase(s) drive these detrimental effects. Methods. Three-month-old WT (C57BL/6) and ADAMTS5-/- mice were chronically exposed to tobacco smoke (four cigarettes/day, 5 day/week for 6 months). ADAMTS-mediated cleavage of disc aggrecan was analyzed by Western blot. Disc total glycosaminoglycan (GAG) content was assessed by dimethyl methylene blue assay and safranin O/fast green histology. Vertebral osteoporosity was measured by microcomputed tomography. Human nucleus pulposus (hNP) cell cultures were also exposed directly to tobacco smoke extract (TSE), a condensate containing the water-soluble compounds inhaled by smokers, to measure ADAMTS5 expression and ADAMTS-mediated cleavage of aggrecan. Activation of nuclear factor (NF)-κB, a family of transcription factors essential for modulating the cellular response to stress, was measured by immunofluorescence assay. Results. Genetic depletion of ADAMTS5 prevented vertebral bone loss, substantially reduced loss of disc GAG content, and completely obviated ADAMTS-mediated proteolysis of disc aggrecan within its interglobular domain (IGD) in mice following exposure to tobacco smoke. hNP cell cultures exposed to TSE also resulted in upregulation of ADAMTS5 protein expression and a concomitant increase in ADAMTS-mediated cleavage within aggrecan IGD. Activation of NF-κB, known to be required for ADAMTS5 gene expression, was observed in both TSE-treated hNP cell cultures and disc tissue of tobacco smoke-exposed mice. Conclusion. The findings demonstrate that ADAMTS5 is the primary aggrecanase mediating smoking-induced disc aggrecanolysis and IDD. Mouse models of chronic tobacco smoking are important and useful for probing the mechanisms of disc aggrecan catabolism and IDD.

AB - Study Design. ADAMTS5-deficient and wild type (WT) mice were chronically exposed to tobacco smoke to investigate effects on intervertebral disc degeneration (IDD). Objective. The aim of this study was to demonstrate a role for ADAMTS5 in mediating tobacco smoking-induced IDD. Summary of Background Data. We previously demonstrated that chronic tobacco smoking causes IDD in mice because, in part, of proteolytic destruction of disc aggrecan. However, it was unknown which matrix proteinase(s) drive these detrimental effects. Methods. Three-month-old WT (C57BL/6) and ADAMTS5-/- mice were chronically exposed to tobacco smoke (four cigarettes/day, 5 day/week for 6 months). ADAMTS-mediated cleavage of disc aggrecan was analyzed by Western blot. Disc total glycosaminoglycan (GAG) content was assessed by dimethyl methylene blue assay and safranin O/fast green histology. Vertebral osteoporosity was measured by microcomputed tomography. Human nucleus pulposus (hNP) cell cultures were also exposed directly to tobacco smoke extract (TSE), a condensate containing the water-soluble compounds inhaled by smokers, to measure ADAMTS5 expression and ADAMTS-mediated cleavage of aggrecan. Activation of nuclear factor (NF)-κB, a family of transcription factors essential for modulating the cellular response to stress, was measured by immunofluorescence assay. Results. Genetic depletion of ADAMTS5 prevented vertebral bone loss, substantially reduced loss of disc GAG content, and completely obviated ADAMTS-mediated proteolysis of disc aggrecan within its interglobular domain (IGD) in mice following exposure to tobacco smoke. hNP cell cultures exposed to TSE also resulted in upregulation of ADAMTS5 protein expression and a concomitant increase in ADAMTS-mediated cleavage within aggrecan IGD. Activation of NF-κB, known to be required for ADAMTS5 gene expression, was observed in both TSE-treated hNP cell cultures and disc tissue of tobacco smoke-exposed mice. Conclusion. The findings demonstrate that ADAMTS5 is the primary aggrecanase mediating smoking-induced disc aggrecanolysis and IDD. Mouse models of chronic tobacco smoking are important and useful for probing the mechanisms of disc aggrecan catabolism and IDD.

KW - ADAMTS5

KW - MMPs

KW - NF-kB

KW - aggrecanases

KW - aggrecanolysis

KW - cellular senescence

KW - genotoxic

KW - inflammatory stress

KW - intervertebral disc

KW - tobacco smoke

UR - http://www.scopus.com/inward/record.url?scp=85020213409&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85020213409&partnerID=8YFLogxK

U2 - 10.1097/BRS.0000000000002258

DO - 10.1097/BRS.0000000000002258

M3 - Article

C2 - 28570296

AN - SCOPUS:85020213409

VL - 42

SP - 1521

EP - 1528

JO - Spine

JF - Spine

SN - 0362-2436

IS - 20

ER -