Systemic inhibition or global deletion of CaMKK2 protects against post-traumatic osteoarthritis

E. Mével; J. A. Shutter; X. Ding; B. T. Mattingly; J. N. Williams; Y. Li; A. Huls; A. V. Kambrath; S. B. Trippel; D. Wagner; M. R. Allen; R. O'Keefe; W. R. Thompson; D. B. Burr; U. Sankar

doi:10.1016/j.joca.2021.09.001

Systemic inhibition or global deletion of CaMKK2 protects against post-traumatic osteoarthritis

E. Mével, J. A. Shutter, X. Ding, B. T. Mattingly, J. N. Williams, Y. Li, A. Huls, A. V. Kambrath, S. B. Trippel, D. Wagner, M. R. Allen, R. O'Keefe, W. R. Thompson, D. B. Burr, U. Sankar

Research output: Contribution to journal › Article › peer-review

Abstract

Objective: To investigate the role of Ca^2+/calmodulin-dependent protein kinase 2 (CaMKK2) in post-traumatic osteoarthritis (PTOA). Methods: Destabilization of the medial meniscus (DMM) or sham surgeries were performed on 10-week-old male wild-type (WT) and Camkk2^−/− mice. Half of the DMM-WT mice and all other cohorts (n = 6/group) received tri-weekly intraperitoneal (i.p.) injections of saline whereas the remaining DMM-WT mice (n = 6/group) received i.p. injections of the CaMKK2 inhibitor STO-609 (0.033 mg/kg body weight) thrice a week. Study was terminated at 8- or 12-weeks post-surgery, and knee joints processed for microcomputed tomography imaging followed by histology and immunohistochemistry. Primary articular chondrocytes were isolated from knee joints of 4–6-day-old WT and Camkk2^−/− mice, and treated with 10 ng/ml interleukin-1β (IL)-1β for 24 or 48 h to investigate gene and protein expression. Results: CaMKK2 levels and activity became elevated in articular chondrocytes following IL-1β treatment or DMM surgery. Inhibition or absence of CaMKK2 protected against DMM-associated destruction of the cartilage, subchondral bone alterations and synovial inflammation. When challenged with IL-1β, chondrocytes lacking CaMKK2 displayed attenuated inflammation, cartilage catabolism, and resistance to suppression of matrix synthesis. IL-1β-treated CaMKK2-null chondrocytes displayed decreased IL-6 production, activation of signal transducer and activator of transcription 3 (Stat3) and matrix metalloproteinase 13 (MMP13), indicating a potential mechanism for the regulation of inflammatory responses in chondrocytes by CaMKK2. Conclusions: Our findings reveal a novel function for CaMKK2 in chondrocytes and highlight the potential for its inhibition as an innovative therapeutic strategy in the prevention of PTOA.

Original language	English
Journal	Osteoarthritis and Cartilage
DOIs	https://doi.org/10.1016/j.joca.2021.09.001
State	Accepted/In press - 2021

Keywords

Adenosine monophosphate dependent protein kinase (AMPK)
Articular chondrocytes
Cacalmodulin-dependent protein kinase kinase 2 (CaMKK2)
Destabilization of medial meniscus (DMM)
IL-6
Interleukin-1β (IL-1β)
Osteoarthritis
Signal transducer and activator of transcription 3 (Stat3)

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10.1016/j.joca.2021.09.001

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Mével, E., Shutter, J. A., Ding, X., Mattingly, B. T., Williams, J. N., Li, Y., Huls, A., Kambrath, A. V., Trippel, S. B., Wagner, D., Allen, M. R., O'Keefe, R., Thompson, W. R., Burr, D. B., & Sankar, U. (Accepted/In press). Systemic inhibition or global deletion of CaMKK2 protects against post-traumatic osteoarthritis. Osteoarthritis and Cartilage. https://doi.org/10.1016/j.joca.2021.09.001

@article{5dff163b578a4863aaaed7299bf7b438,

title = "Systemic inhibition or global deletion of CaMKK2 protects against post-traumatic osteoarthritis",

abstract = "Objective: To investigate the role of Ca2+/calmodulin-dependent protein kinase 2 (CaMKK2) in post-traumatic osteoarthritis (PTOA). Methods: Destabilization of the medial meniscus (DMM) or sham surgeries were performed on 10-week-old male wild-type (WT) and Camkk2−/− mice. Half of the DMM-WT mice and all other cohorts (n = 6/group) received tri-weekly intraperitoneal (i.p.) injections of saline whereas the remaining DMM-WT mice (n = 6/group) received i.p. injections of the CaMKK2 inhibitor STO-609 (0.033 mg/kg body weight) thrice a week. Study was terminated at 8- or 12-weeks post-surgery, and knee joints processed for microcomputed tomography imaging followed by histology and immunohistochemistry. Primary articular chondrocytes were isolated from knee joints of 4–6-day-old WT and Camkk2−/− mice, and treated with 10 ng/ml interleukin-1β (IL)-1β for 24 or 48 h to investigate gene and protein expression. Results: CaMKK2 levels and activity became elevated in articular chondrocytes following IL-1β treatment or DMM surgery. Inhibition or absence of CaMKK2 protected against DMM-associated destruction of the cartilage, subchondral bone alterations and synovial inflammation. When challenged with IL-1β, chondrocytes lacking CaMKK2 displayed attenuated inflammation, cartilage catabolism, and resistance to suppression of matrix synthesis. IL-1β-treated CaMKK2-null chondrocytes displayed decreased IL-6 production, activation of signal transducer and activator of transcription 3 (Stat3) and matrix metalloproteinase 13 (MMP13), indicating a potential mechanism for the regulation of inflammatory responses in chondrocytes by CaMKK2. Conclusions: Our findings reveal a novel function for CaMKK2 in chondrocytes and highlight the potential for its inhibition as an innovative therapeutic strategy in the prevention of PTOA.",

keywords = "Adenosine monophosphate dependent protein kinase (AMPK), Articular chondrocytes, Cacalmodulin-dependent protein kinase kinase 2 (CaMKK2), Destabilization of medial meniscus (DMM), IL-6, Interleukin-1β (IL-1β), Osteoarthritis, Signal transducer and activator of transcription 3 (Stat3)",

author = "E. M{\'e}vel and Shutter, {J. A.} and X. Ding and Mattingly, {B. T.} and Williams, {J. N.} and Y. Li and A. Huls and Kambrath, {A. V.} and Trippel, {S. B.} and D. Wagner and Allen, {M. R.} and R. O'Keefe and Thompson, {W. R.} and Burr, {D. B.} and U. Sankar",

note = "Funding Information: Research reported in this manuscript was supported by the National Institute Of Arthritis And Musculoskeletal And Skin Diseases of the National Institutes of Health under Award Numbers R01AR076477 and R01AR068332 (both to US); and an Indiana Center for Musculoskeletal Heath (ICMH) Pilot Grant Award to US. This work was also supported by funds from the Indiana Clinical and Translational Sciences Institute funded in part by Award Number UL1TR001108 from the National Institutes of Health, National Center for Advancing Translational Sciences , Clinical and Translational Sciences Award to EM. The 1172 μCT system was supported by an S10 grant (S10-RR023710) to MRA. JN was supported through a Comprehensive Musculoskeletal T32 Training Program from NIAMS/NIH ( AR065971 ). JAS is supported by the Moenkhaus fellowship from the Department of Anatomy, Cell Biology & Physiology at IUSM . The study sponsors were not involved in any aspects of this study including study design, collection, analysis and interpretation of data; or in the writing of the manuscript, and in the decision to submit the manuscript for publication. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Publisher Copyright: {\textcopyright} 2021 Osteoarthritis Research Society International",

year = "2021",

doi = "10.1016/j.joca.2021.09.001",

language = "English",

journal = "Osteoarthritis and Cartilage",

issn = "1063-4584",

}

Systemic inhibition or global deletion of CaMKK2 protects against post-traumatic osteoarthritis. / Mével, E.; Shutter, J. A.; Ding, X.; Mattingly, B. T.; Williams, J. N.; Li, Y.; Huls, A.; Kambrath, A. V.; Trippel, S. B.; Wagner, D.; Allen, M. R.; O'Keefe, R.; Thompson, W. R.; Burr, D. B.; Sankar, U.

In: Osteoarthritis and Cartilage, 2021.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Systemic inhibition or global deletion of CaMKK2 protects against post-traumatic osteoarthritis

AU - Mével, E.

AU - Shutter, J. A.

AU - Ding, X.

AU - Mattingly, B. T.

AU - Williams, J. N.

AU - Li, Y.

AU - Huls, A.

AU - Kambrath, A. V.

AU - Trippel, S. B.

AU - Wagner, D.

AU - Allen, M. R.

AU - O'Keefe, R.

AU - Thompson, W. R.

AU - Burr, D. B.

AU - Sankar, U.

N1 - Funding Information: Research reported in this manuscript was supported by the National Institute Of Arthritis And Musculoskeletal And Skin Diseases of the National Institutes of Health under Award Numbers R01AR076477 and R01AR068332 (both to US); and an Indiana Center for Musculoskeletal Heath (ICMH) Pilot Grant Award to US. This work was also supported by funds from the Indiana Clinical and Translational Sciences Institute funded in part by Award Number UL1TR001108 from the National Institutes of Health, National Center for Advancing Translational Sciences , Clinical and Translational Sciences Award to EM. The 1172 μCT system was supported by an S10 grant (S10-RR023710) to MRA. JN was supported through a Comprehensive Musculoskeletal T32 Training Program from NIAMS/NIH ( AR065971 ). JAS is supported by the Moenkhaus fellowship from the Department of Anatomy, Cell Biology & Physiology at IUSM . The study sponsors were not involved in any aspects of this study including study design, collection, analysis and interpretation of data; or in the writing of the manuscript, and in the decision to submit the manuscript for publication. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Publisher Copyright: © 2021 Osteoarthritis Research Society International

PY - 2021

Y1 - 2021

N2 - Objective: To investigate the role of Ca2+/calmodulin-dependent protein kinase 2 (CaMKK2) in post-traumatic osteoarthritis (PTOA). Methods: Destabilization of the medial meniscus (DMM) or sham surgeries were performed on 10-week-old male wild-type (WT) and Camkk2−/− mice. Half of the DMM-WT mice and all other cohorts (n = 6/group) received tri-weekly intraperitoneal (i.p.) injections of saline whereas the remaining DMM-WT mice (n = 6/group) received i.p. injections of the CaMKK2 inhibitor STO-609 (0.033 mg/kg body weight) thrice a week. Study was terminated at 8- or 12-weeks post-surgery, and knee joints processed for microcomputed tomography imaging followed by histology and immunohistochemistry. Primary articular chondrocytes were isolated from knee joints of 4–6-day-old WT and Camkk2−/− mice, and treated with 10 ng/ml interleukin-1β (IL)-1β for 24 or 48 h to investigate gene and protein expression. Results: CaMKK2 levels and activity became elevated in articular chondrocytes following IL-1β treatment or DMM surgery. Inhibition or absence of CaMKK2 protected against DMM-associated destruction of the cartilage, subchondral bone alterations and synovial inflammation. When challenged with IL-1β, chondrocytes lacking CaMKK2 displayed attenuated inflammation, cartilage catabolism, and resistance to suppression of matrix synthesis. IL-1β-treated CaMKK2-null chondrocytes displayed decreased IL-6 production, activation of signal transducer and activator of transcription 3 (Stat3) and matrix metalloproteinase 13 (MMP13), indicating a potential mechanism for the regulation of inflammatory responses in chondrocytes by CaMKK2. Conclusions: Our findings reveal a novel function for CaMKK2 in chondrocytes and highlight the potential for its inhibition as an innovative therapeutic strategy in the prevention of PTOA.

AB - Objective: To investigate the role of Ca2+/calmodulin-dependent protein kinase 2 (CaMKK2) in post-traumatic osteoarthritis (PTOA). Methods: Destabilization of the medial meniscus (DMM) or sham surgeries were performed on 10-week-old male wild-type (WT) and Camkk2−/− mice. Half of the DMM-WT mice and all other cohorts (n = 6/group) received tri-weekly intraperitoneal (i.p.) injections of saline whereas the remaining DMM-WT mice (n = 6/group) received i.p. injections of the CaMKK2 inhibitor STO-609 (0.033 mg/kg body weight) thrice a week. Study was terminated at 8- or 12-weeks post-surgery, and knee joints processed for microcomputed tomography imaging followed by histology and immunohistochemistry. Primary articular chondrocytes were isolated from knee joints of 4–6-day-old WT and Camkk2−/− mice, and treated with 10 ng/ml interleukin-1β (IL)-1β for 24 or 48 h to investigate gene and protein expression. Results: CaMKK2 levels and activity became elevated in articular chondrocytes following IL-1β treatment or DMM surgery. Inhibition or absence of CaMKK2 protected against DMM-associated destruction of the cartilage, subchondral bone alterations and synovial inflammation. When challenged with IL-1β, chondrocytes lacking CaMKK2 displayed attenuated inflammation, cartilage catabolism, and resistance to suppression of matrix synthesis. IL-1β-treated CaMKK2-null chondrocytes displayed decreased IL-6 production, activation of signal transducer and activator of transcription 3 (Stat3) and matrix metalloproteinase 13 (MMP13), indicating a potential mechanism for the regulation of inflammatory responses in chondrocytes by CaMKK2. Conclusions: Our findings reveal a novel function for CaMKK2 in chondrocytes and highlight the potential for its inhibition as an innovative therapeutic strategy in the prevention of PTOA.

KW - Adenosine monophosphate dependent protein kinase (AMPK)

KW - Articular chondrocytes

KW - Cacalmodulin-dependent protein kinase kinase 2 (CaMKK2)

KW - Destabilization of medial meniscus (DMM)

KW - IL-6

KW - Interleukin-1β (IL-1β)

KW - Osteoarthritis

KW - Signal transducer and activator of transcription 3 (Stat3)

UR - http://www.scopus.com/inward/record.url?scp=85115421623&partnerID=8YFLogxK

U2 - 10.1016/j.joca.2021.09.001

DO - 10.1016/j.joca.2021.09.001

M3 - Article

C2 - 34506942

AN - SCOPUS:85115421623

JO - Osteoarthritis and Cartilage

JF - Osteoarthritis and Cartilage

SN - 1063-4584

ER -

Systemic inhibition or global deletion of CaMKK2 protects against post-traumatic osteoarthritis

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Keywords

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