Skeletal modeling represents the shaping of bone as the skeleton develops both pre- and post-natally in order to meet the needs of structural integrity and mechanical competence. Like remodeling, modeling requires coordination of osteoblasts (Obl), osteocytes (Ocy) and osteoclasts (Ocl). Unlike remodeling however, modeling osteoclasts and osteoblasts are not anatomically tethered but respectively resorb and form bone in locations which yield appropriate movement of the skeleton through space. During the tenure of this project, we demonstrated that bone forming cells communicate via gap junctions formed primarily by connexin43 (Cx43). The presence of cortical bone abnormalities in mice with Cx43 gene (Gja1) ablation in cells of the osteogenic lineage, and the fact that Gja1 mutations cause the human disease oculodentodigital dysplasia (ODDD), characterized by craniofacial and skeletal abnormalities, substantiate that skeletal homeostasis requires Cx43. In the last funding cycle, we made the unexpected observation that conditional Gja1 inactivation or induction of an ODDD Gja1 mutation in osteogenic cells primarily impairs cortical bone structure, leading to a larger total area, but decreased cortical thickness and compromised bone strength. This phenotype reflects increased endocortical resorption and periosteal apposition, as well as a hypomineralized and structurally abnormal bone matrix, associated with down-regulation of osteoprotegerin (Opg) and Sost in Obl/Ocy. Lack of Cx43 also alters the sensitivity of cortical bone to mechanical loading and unloading in an envelope-specific fashion. Hence, Cx43 in osteogenic cells controls envelope-specific formation and resorption of cortical bone, thus dictating its size, shape, biomechanical properties and responsiveness to mechanical stimuli. We therefore hypothesize that Cx43 is a key modulator of cortical bone modeling in the adult skeleton, and propose the following Specific Aims: 1) Mechanisms of Cx43 regulation of endocortical bone resorption, which will test the hypothesis that Cx43 regulates endocortical bone resorption via modulation of osteoprotegerin (Opg) expression by Obl and/or Ocy;2) Mechanisms of Cx43 regulation of periosteal bone formation, which will test the hypothesis that Cx43 regulates periosteal bone formation, in part via modulation of Wnt signals;3) Mechanical regulation of periosteal bone formation via Cx43, which will test the hypothesis that Cx43 modulates cortical bone responses to mechanical loading independently of bone architecture, via cell autonomous actions (Obl specific) and paracrine mechanisms (via the Ocy). Cortical bone (re)modeling is understudied but its importance in bone homeostasis and in the adaptive responses to mechanical factors is critical for maintaining bone strength and resistance to fractures. The proposed experiments will use genetic mouse models wherein Gja1 is selectively ablated or mutated at different stages of osteogenesis. Results will establish the mechanisms by which Cx43 modulates cortical bone and how such mechanisms cause the skeletal abnormalities of ODDD, findings that may be translated to clinical settings and lead to pharmacologic targeting.

Public Health Relevance

Cortical bone is critical for optimal bone strength and resistance to fractures;however, little is known about how the bone cortex is maintained in the adult skeleton. We have discovered that the gap junction protein, Cx43 controls both endocortical bone resorption and periosteal formation, and alters the effect of mechanical load on the cortex. The proposed experiments will establish the mechanisms by which Cx43 modulates cortical bone and how such mechanisms lead to the skeletal abnormalities of oculodentodigital dysplasia, a disease caused by Cx43 gene mutations. Results would help us devise new therapeutic strategies to improve cortical bone structure and strength.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
2R01AR041255-21A1
Application #
8589808
Study Section
Skeletal Biology Development and Disease Study Section (SBDD)
Program Officer
Chen, Faye H
Project Start
1992-01-01
Project End
2018-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
21
Fiscal Year
2013
Total Cost
$323,000
Indirect Cost
$110,500
Name
Washington University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Stains, Joseph P; Civitelli, Roberto (2016) Connexins in the skeleton. Semin Cell Dev Biol 50:31-9
Stains, Joseph P; Civitelli, Roberto (2016) A Functional Assay to Assess Connexin 43-Mediated Cell-to-Cell Communication of Second Messengers in Cultured Bone Cells. Methods Mol Biol 1437:193-201
Shen, Hua; Grimston, Susan; Civitelli, Roberto et al. (2015) Deletion of connexin43 in osteoblasts/osteocytes leads to impaired muscle formation in mice. J Bone Miner Res 30:596-605
Chang, Kyung Hee; Sengupta, Amitava; Nayak, Ramesh C et al. (2014) p62 is required for stem cell/progenitor retention through inhibition of IKK/NF-κB/Ccl4 signaling at the bone marrow macrophage-osteoblast niche. Cell Rep 9:2084-97
Stains, Joseph P; Watkins, Marcus P; Grimston, Susan K et al. (2014) Molecular mechanisms of osteoblast/osteocyte regulation by connexin43. Calcif Tissue Int 94:55-67
Grimston, Susan K; Watkins, Marcus P; Stains, Joseph P et al. (2013) Connexin43 modulates post-natal cortical bone modeling and mechano-responsiveness. Bonekey Rep 2:446
Watkins, Marcus P; Norris, Jin Yi; Grimston, Susan K et al. (2012) Bisphosphonates improve trabecular bone mass and normalize cortical thickness in ovariectomized, osteoblast connexin43 deficient mice. Bone 51:787-94
Gonzalez-Nieto, Daniel; Li, Lina; Kohler, Anja et al. (2012) Connexin-43 in the osteogenic BM niche regulates its cellular composition and the bidirectional traffic of hematopoietic stem cells and progenitors. Blood 119:5144-54
Grimston, Susan K; Watkins, Marcus P; Brodt, Michael D et al. (2012) Enhanced periosteal and endocortical responses to axial tibial compression loading in conditional connexin43 deficient mice. PLoS One 7:e44222
Watkins, Marcus; Grimston, Susan K; Norris, Jin Yi et al. (2011) Osteoblast connexin43 modulates skeletal architecture by regulating both arms of bone remodeling. Mol Biol Cell 22:1240-51

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