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

Osteoarthritis Cartilage. 2022 Jan;30(1):124-136. doi: 10.1016/j.joca.2021.09.001. Epub 2021 Sep 17.

Authors

E Mével¹, J A Shutter², X Ding³, B T Mattingly⁴, J N Williams⁵, Y Li⁶, A Huls⁷, A V Kambrath⁸, S B Trippel⁹, D Wagner¹⁰, M R Allen¹¹, R O'Keefe¹², W R Thompson¹³, D B Burr¹⁴, U Sankar¹⁵

Affiliations

¹ Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
² Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
³ Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
⁴ Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
⁵ Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
⁶ Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
⁷ Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
⁸ Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Department of Orthopaedic Surgery, Indiana University School of Medicine, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
⁹ Department of Orthopaedic Surgery, Indiana University School of Medicine, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
¹⁰ Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Department of Orthopaedic Surgery, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Department of Mechanical and Energy Engineering, School of Engineering and Technology, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
¹¹ Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Department of Orthopaedic Surgery, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
¹² Department of Orthopaedic Surgery, Washington University School of Medicine, St. Louis, MO, 63110, USA. Electronic address: [email protected].
¹³ Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Department of Physical Therapy, School of Health and Rehabilitation Sciences, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
¹⁴ Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, 46202, USA. Electronic address: [email protected].
¹⁵ Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA; Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, 46202, USA. Electronic address: [email protected].

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.

Keywords: Adenosine monophosphate dependent protein kinase (AMPK); Articular chondrocytes; Ca(2+/)calmodulin-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).

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Benzimidazoles / therapeutic use*
Calcium-Calmodulin-Dependent Protein Kinase Kinase / antagonists & inhibitors*
Calcium-Calmodulin-Dependent Protein Kinase Kinase / physiology*
Cartilage, Articular / injuries*
Male
Mice
Naphthalimides / therapeutic use*
Osteoarthritis / etiology*
Osteoarthritis / prevention & control*
Wounds and Injuries / complications

Substances

Benzimidazoles
Naphthalimides
STO 609
Calcium-Calmodulin-Dependent Protein Kinase Kinase
Camkk2 protein, mouse

Abstract

Publication types

MeSH terms

Substances

Grants and funding