Inhibition of CaMKK2 reverses age-associated decline in bone mass.
Decline in bone formation is a major contributing factor to the loss of bone mass associated with aging. We previously showed that the genetic ablation of the tissue-restricted and multifunctional Ca(2+)/calmodulin (CaM)-dependent protein kinase kinase 2 (CaMKK2) stimulates trabecular bone mass accrual, mainly by promoting anabolic pathways and inhibiting catabolic pathways of bone remodeling. In this study, we investigated whether inhibition of this kinase using its selective cell-permeable inhibitor STO-609 will stimulate bone formation in 32 week old male WT mice and reverse age-associated of decline in bone volume and strength. Tri-weekly intraperitoneal injections of saline or STO-609 (10 μM) were performed for six weeks followed by metabolic labeling with calcein and alizarin red. New bone formation was assessed by dynamic histomorphometry whereas micro-computed tomography was employed to measure trabecular bone volume, microarchitecture and femoral mid-shaft geometry. Cortical and trabecular bone biomechanical properties were assessed using three-point bending and punch compression methods respectively. Our results reveal that as they progress from 12 to 32 weeks of age, WT mice sustain a significant decline in trabecular bone volume, microarchitecture and strength as well as cortical bone strength. However, treatment of the 32 week old WT mice with STO-609 stimulated apposition of new bone and completely reversed the age-associated decrease in bone volume, quality, as well as trabecular and cortical bone strength. We also observed that regardless of age, male Camkk2(-/-) mice possessed significantly elevated trabecular bone volume, microarchitecture and compressive strength as well as cortical bone strength compared to age-matched WT mice, implying that the chronic loss of this kinase attenuates age-associated decline in bone mass. Further, whereas STO-609 treatment and/or the absence of CaMKK2 significantly enhanced the femoral mid-shaft geometry, the mid-shaft cortical wall thickness and material bending stress remained similar among the cohorts, implying that regardless of treatment, the material properties of the bone remain similar. Thus, our cumulative results provide evidence for the pharmacological inhibition of CaMKK2 as a bone anabolic strategy in combating age-associated osteoporosis.
Department of Pharmacology and Toxicology, University of Louisville School of Medicine, Louisville, KY, USA.
Zachary J
ZJ
Pritchard
James Graham Brown Cancer Center, University of Louisville School of Medicine, Louisville, KY, USA.
Rachel L
RL
Cary
Department of Medicine and Pathology, Washington University School of Medicine, St. Louis, MO, USA.
Chang
C
Yang
Department of Medicine and Pathology, Washington University School of Medicine, St. Louis, MO, USA.
Deborah V
DV
Novack
Department of Orthopaedic Surgery, University of Louisville School of Medicine, Louisville, KY, USA; Department of Bioengineering, University of Louisville Speed School of Engineering, Louisville, KY, USA. Electronic address: michael.voor@louisville.edu.
Michael J
MJ
Voor
Department of Pharmacology and Toxicology, University of Louisville School of Medicine, Louisville, KY, USA; James Graham Brown Cancer Center, University of Louisville School of Medicine, Louisville, KY, USA; Department of Anatomy and Cell Biology, Indiana University School of Medicine, Indianapolis, IN, USA. Electronic address: usankar@iupui.edu.
Uma
U
Sankar
D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology*; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase / Q000037:antagonists & inhibitors*; D019245:Compressive Strength; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005269:Femur / Q000000981:diagnostic imaging; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D053644:Naphthalimides / Q000494:pharmacology*; D010012:Osteogenesis / Q000187:drug effects*; D010024:Osteoporosis / Q000201:enzymology / Q000473:pathology; D055114:X-Ray Microtomography
D001562:Benzimidazoles; D004791:Enzyme Inhibitors; D053644:Naphthalimides; C458525:STO 609; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase; C517700:Camkk2 protein, mouse
Bone volume; Ca(2+)/calmodulin (CaM)-dependent protein kinase kinase 2; Microarchitecture; STO-609; Three-point bending; Trabecular punch compression