Calcitonin (CT) is the most potent and rapidly acting known inhibitor of bone resorption that targets the osteoclast (OC) directly. CT was the first therapeutic agent used to inhibit excessive bone resorption in osteoporosis and other conditions. It is still used, and new formulations for treating osteoarthritis and osteoporosis are currently in development, despite the fact that a reported loss of efficacy with time has somewhat limited clinical use. Thus, new information about the mechanisms by which CT affects OC function has potential therapeutic importance. In vitro, CT rapidly inhibits OC motility and disrupts the podosome belt, the cytoskeletal attachment feature of mature resorbing OCs, accounting for much of CT's inhibition of bone resorption. During the past funding period we showed that CT inhibits OC spreading, motility and bone resorption in large part by inhibiting the protease calpain, and that inhibiting calpain also promotes the recycling of the CT receptor to the cell surface by filamin. These results are a major breakthrough in understanding how CT regulates OC cytoskeleton organization and podosome function. This renewal application seeks to characterize the calpain-dependent mechanisms of CT action, focusing on how CT regulates calpain activity in OCs and how calpain regulates OC attachment and motility, which will provide novel insights into the regulation of osteoclast function in bone remodeling.
The Specific Aims of this proposal are therefore: 1. To further elucidate the mechanisms by which calpain modulates podosome function, cytoskeletal organization, cell adhesion and motility, specifically focusing on proteins (cortactin, talin, Pyk2, filamin) that we or others have identified as calpain substrates that play key functional roles in adhesion complexes in osteoclasts; 2. To further characterize the coupling of the CTR to calpain in OCs and the calpain- independent effects of CTR-induced signaling effectors on the function of podosome components; 3. To further characterize the molecular basis of the CTR-filamin interaction, and determine how that interaction affects CTR function, and more specifically, CT's regulation of OC attachment and motility. These studies will reveal previously unappreciated features of CT's mechanisms of inhibiting bone resorption, and thereby possibly increase CT's therapeutic potential by creating the possibility of rapidly and potently inhibiting bone resorption while avoiding the loss of response.

Public Health Relevance

The hormone calcitonin is the most potent and rapidly acting known inhibitor of bone resorption. It was the first therapeutic agent used to inhibit excessive bone resorption in osteoporosis and other conditions and is still used, despite a reported loss of efficacy with time. Calcitonin rapidly inhibits the motility of osteoclasts, the cells that resorb mineralized bone, in part by disrupting the structures that the osteoclast uses to attach to the bone surface. The purpose of this application is to better understand the molecular mechanisms of calcitonin action, in particular the reason that the inhibition of the proteolytic enzyme calpain by calcitonin disrupts the osteoclast attachment structures and how calpain regulates osteoclast attachment and motility. This study will provide novel insights into the regulation of osteoclast function in bone remodeling, and thereby possibly increase the development of calcitonin's therapeutic potential.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR049879-10
Application #
8293432
Study Section
Skeletal Biology Structure and Regeneration Study Section (SBSR)
Program Officer
Chen, Faye H
Project Start
2003-06-01
Project End
2014-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
10
Fiscal Year
2012
Total Cost
$362,459
Indirect Cost
$148,619
Name
Harvard University
Department
Dentistry
Type
Schools of Dentistry
DUNS #
047006379
City
Boston
State
MA
Country
United States
Zip Code
02115
Coury, Fabienne; Zenger, Serhan; Stewart, Andrew K et al. (2013) SLC4A2-mediated Cl-/HCO3- exchange activity is essential for calpain-dependent regulation of the actin cytoskeleton in osteoclasts. Proc Natl Acad Sci U S A 110:2163-8
Marzia, Marilena; Chiusaroli, Riccardo; Neff, Lynn et al. (2006) Calpain is required for normal osteoclast function and is down-regulated by calcitonin. J Biol Chem 281:9745-54
Seck, Thomas; Pellegrini, Maria; Florea, Ana Maria et al. (2005) The delta e13 isoform of the calcitonin receptor forms a six-transmembrane domain receptor with dominant-negative effects on receptor surface expression and signaling. Mol Endocrinol 19:2132-44
Beaudreuil, J; Balasubramanian, Sundaravadivel; Chenais, J et al. (2004) Molecular characterization of two novel isoforms of the human calcitonin receptor. Gene 343:143-51