Calcium-Sensing Receptors and Growth Plate Development

Summary

Principal Investigator: Wenhan Chang
Affiliation: University of California
Country: USA
Abstract: DESCRIPTION (provided by applicant): Growth plate development is a critical step in endochondral bone formation. Delayed cell differentiation in the growth plate can produce chondrodysplasia, hypophosphatasia, rickets, and growth retardation. The orderly progression of chondrocyte differentiation is maintained by hormones, nutritional components, and autocrine and paracrine factors produced by chondrocytes. The concentration of Ca2+ in the extracellular fluid ([Ca2+]e) plays a key role in growth plate cartilage development. Certain forms of childhood rickets are due to extremely low Ca2+ intakes. The bones and cartilage in rickets are soft and deformed and growth is stunted. We found that raising the [Ca2+]e directly promotes the terminal differentiation and mineralization of cultured mouse growth plate chondrocytes (mGPCs). Like Ca2+, insulin-like growth factor-1 (IGF1) promotes GPC differentiation. High Ca2+ also increases IGF1/IGF1R signaling capacity in mGPCs, suggesting that IGF1R-mediated signaling may be involved in the high [Ca2+]e-induces cell differentiation. The signaling mechanisms responsible for the effects of Ca2+ in the growth plate have not been defined. The Ca2+-sensing receptor (CaR) is strongly expressed in mGPCs. Altering CaR expression and function profoundly impacts the differentiation of these cells. To determine whether the CaR is responsible for extracellular Ca2+-sensing by cartilage in vivo, we generated mice [CartCaR(-/-)] with tissue-specific knockout of the CaR in chondrocytes by a Cre-lox recombination technique. These mice die in gestation with malformed skeletons. We also developed a tamoxifen-inducible, chondrocyte-specific CaR knockout mouse [(Tam-CartCaR(-/-)]. These mice grow poorly during their postnatal life and have abnormally expanded growth plates with delayed mineralization in their hypertrophic zones -- features seen in rickets. These data strongly support a role for the CaR in growth plate development. We will use these models to address the hypothesis that a high [Ca2+]e by activating CaRs stimulates ERK1/2 and PLC activity, increases the intracellular [Ca2+], enhances the expression of and signaling by IGF1 and the IGF1R, and promotes the terminal differentiation of GPCs. We propose the following AIMs: (1) to determine the role of the CaR at specific stages of growth plate development by inducing CaR gene deletion with Tam at timed intervals before and after birth;(2) to determine whether a high [Ca2+]e alters chondrocyte function by regulating the expression of and signaling by IGF1 and the IGF1R in chondrocytes;(3) to assess the contribution of IGF1 and IGF1R-mediated signaling to the responses of GPCs to changes in the [Ca2+]e;and (4) to identify the high [Ca2+]e-induced signaling responses that are mediated by the CaR and are responsible for changes in the growth, survival, and differentiation of chondrocytes. Successful completion of these studies will provide insights into how the CaR interacts with local IGF1/IGF1R signaling pathway to modulate GPC differentiation and growth plate development. Our studies will advance the field of CaR biology and may provide a potential pharmaceutical target for disorders of growth plate function in childhood. PUBLIC HEALTH RELEVANCE: Growth plate cartilage, which is located at the ends of the bones, allows them to grow bigger. Chondrocytes from growth plate cartilage are capable of sensing changes in the concentrations of Ca2+ in their surroundings and adjusting their activities accordingly. Changes in the concentrations of Ca2+ near growth plate chondrocytes influence their whole program of gene expression, matrix protein synthesis, and mineral deposition. This proposal will investigate how Ca2+ acts on these chondrocytes and will help us understand how to treat disorders of growth and poor mineralization like rickets.
Funding Period: ----------------2002 - ---------------2013-
more information: NIH RePORT

Top Publications

  1. ncbi Insulin-like growth factor-I is essential for embryonic bone development
    Yongmei Wang
    Department of Medicine, Endocrine Unit, 111N, Veterans Affairs Medical Center, 4150 Clement Street, San Francisco, California 94121, USA
    Endocrinology 147:4753-61. 2006
  2. ncbi The extracellular calcium-sensing receptor, CaSR, in fetal development
    Daniela Riccardi
    School of Biosciences, Cardiff University, Cardiff, UK
    Best Pract Res Clin Endocrinol Metab 27:443-53. 2013
  3. pmc Mammary-specific ablation of the calcium-sensing receptor during lactation alters maternal calcium metabolism, milk calcium transport, and neonatal calcium accrual
    Ramanaiah Mamillapalli
    Section of Endocrinology and Metabolism, Department of Internal Medicine, Yale University School of Medicine, TAC S131, Box 208020, New Haven, Connecticut 06520 8020, USA
    Endocrinology 154:3031-42. 2013
  4. pmc Sex and age modify biochemical and skeletal manifestations of chronic hyperparathyroidism by altering target organ responses to Ca2+ and parathyroid hormone in mice
    Zhiqiang Cheng
    Endocrine Research Unit, Department of Veterans Affairs Medical Center, Department of Medicine, University of California, San Francisco, CA, USA
    J Bone Miner Res 28:1087-100. 2013
  5. pmc Ablation of the calcium-sensing receptor in keratinocytes impairs epidermal differentiation and barrier function
    Chia Ling Tu
    Endocrine Unit, Veterans Affairs Medical Center, University of California, San Francisco, San Francisco, California 94121, USA
    J Invest Dermatol 132:2350-9. 2012
  6. pmc Ephrin B2/EphB4 Mediates the Actions of IGF-I Signaling in Regulating Endochondral Bone Formation
    Yongmei Wang
    Endocrine Unit, University of California, Veterans Affairs Medical Center, San Francisco, CA, USA
    J Bone Miner Res 29:1900-13. 2014
  7. pmc Assessing constitutive activity of extracellular calcium-sensing receptors in vitro and in bone
    Wenhan Chang
    Endocrine Research Unit, Department of Veterans Affairs Medical Center, Department of Medicine, University of California, San Francisco, California, USA
    Methods Enzymol 484:253-66. 2010
  8. pmc The extracellular calcium-sensing receptor (CaSR) is a critical modulator of skeletal development
    Wenhan Chang
    Endocrine Research Unit, Department of Veterans Affairs Medical Center, Department of Medicine, University of California, San Francisco, CA 94121, USA
    Sci Signal 1:ra1. 2008
  9. ncbi Inactivation of the calcium sensing receptor inhibits E-cadherin-mediated cell-cell adhesion and calcium-induced differentiation in human epidermal keratinocytes
    Chia Ling Tu
    Endocrine Unit, Veteran Affairs Medical Center, University of California, San Francisco, California 94121, USA
    J Biol Chem 283:3519-28. 2008
  10. ncbi The role of the calcium sensing receptor in regulating intracellular calcium handling in human epidermal keratinocytes
    Chia Ling Tu
    Endocrine Unit, Department of Medicine, Veteran Affairs Medical Center and University of California, San Francisco, California 94121, USA
    J Invest Dermatol 127:1074-83. 2007

Scientific Experts

  • OSVALDO A REY
  • Qing Dong
  • Chia Ling Tu
  • Wenhan Chang
  • Yongmei Wang
  • Zhiqiang Cheng
  • Tsui Hua Chen
  • Daniel D Bikle
  • Dolores Shoback
  • Daniel Bikle
  • Hashem Z Elalieh
  • Melita M Dvorak
  • Ramanaiah Mamillapalli
  • Daniela Riccardi
  • Nathan Liang
  • Melita M Dvorak-Ewell
  • Dolores M Shoback
  • Caitlin Garvey
  • Luis Rodriguez
  • Bernhard Bettler
  • Martin Gassmann
  • Marta Margeta
  • Chak Fong
  • Alicia Menendez
  • Christian Santa Maria
  • Edward Brown
  • John Wysolmerski
  • Hanson Ho
  • Fuqing Song
  • Alfred Li
  • Joshua VanHouten
  • Pamela Dann
  • Sarah C Brennan
  • Michael You
  • Thomas L Clemens
  • Betty Liu
  • Susan Mackem
  • Eiichiro Nakamura
  • Minh Thanh Nguyen
  • Melita Dvorak
  • Lily Y Jan
  • Benjamin Orwoll
  • Takeshi Sakata
  • Bernard P Halloran
  • Shigeki Nishida
  • Steven B Doty

Detail Information

Publications19

  1. ncbi Insulin-like growth factor-I is essential for embryonic bone development
    Yongmei Wang
    Department of Medicine, Endocrine Unit, 111N, Veterans Affairs Medical Center, 4150 Clement Street, San Francisco, California 94121, USA
    Endocrinology 147:4753-61. 2006
    ..Our results indicate that IGF-I plays an important role in skeletal development by promoting chondrocyte proliferation and maturation while inhibiting apoptosis to form bones of appropriate size and strength...
  2. ncbi The extracellular calcium-sensing receptor, CaSR, in fetal development
    Daniela Riccardi
    School of Biosciences, Cardiff University, Cardiff, UK
    Best Pract Res Clin Endocrinol Metab 27:443-53. 2013
    ..In this article, we present evidence for a role of the CaSR in the control of skeletal development, and how fetal hypercalcemia, acting through the CaSR, regulates lung development. ..
  3. pmc Mammary-specific ablation of the calcium-sensing receptor during lactation alters maternal calcium metabolism, milk calcium transport, and neonatal calcium accrual
    Ramanaiah Mamillapalli
    Section of Endocrinology and Metabolism, Department of Internal Medicine, Yale University School of Medicine, TAC S131, Box 208020, New Haven, Connecticut 06520 8020, USA
    Endocrinology 154:3031-42. 2013
    ..These results demonstrate that the mammary gland CaSR coordinates maternal bone and calcium metabolism, calcium transport into milk, and neonatal calcium accrual during lactation. ..
  4. pmc Sex and age modify biochemical and skeletal manifestations of chronic hyperparathyroidism by altering target organ responses to Ca2+ and parathyroid hormone in mice
    Zhiqiang Cheng
    Endocrine Research Unit, Department of Veterans Affairs Medical Center, Department of Medicine, University of California, San Francisco, CA, USA
    J Bone Miner Res 28:1087-100. 2013
    ..Restraining the ability of the PTG to upregulate CaSRs by heterozygous gene deletion contributes to biochemical and skeletal manifestations of HPT, especially in aging females...
  5. pmc Ablation of the calcium-sensing receptor in keratinocytes impairs epidermal differentiation and barrier function
    Chia Ling Tu
    Endocrine Unit, Veterans Affairs Medical Center, University of California, San Francisco, San Francisco, California 94121, USA
    J Invest Dermatol 132:2350-9. 2012
    ..3%) diet. Deleting CaR perturbs the epidermal Ca(2+) gradient and impairs keratinocyte differentiation and permeability barrier homeostasis, indicating a key role for the CaR in normal epidermal development...
  6. pmc Ephrin B2/EphB4 Mediates the Actions of IGF-I Signaling in Regulating Endochondral Bone Formation
    Yongmei Wang
    Endocrine Unit, University of California, Veterans Affairs Medical Center, San Francisco, CA, USA
    J Bone Miner Res 29:1900-13. 2014
    ..Our data indicate that IGF-I/IGF-IR signaling promotes OB, OCL, and chondrocyte differentiation via ephrin B2/EphB4 mediated cell-cell communication. © 2014 American Society for Bone and Mineral Research. ..
  7. pmc Assessing constitutive activity of extracellular calcium-sensing receptors in vitro and in bone
    Wenhan Chang
    Endocrine Research Unit, Department of Veterans Affairs Medical Center, Department of Medicine, University of California, San Francisco, California, USA
    Methods Enzymol 484:253-66. 2010
    ..These studies indicate that this approach can be readily adapted to assess CaSR actions in other cell systems...
  8. pmc The extracellular calcium-sensing receptor (CaSR) is a critical modulator of skeletal development
    Wenhan Chang
    Endocrine Research Unit, Department of Veterans Affairs Medical Center, Department of Medicine, University of California, San Francisco, CA 94121, USA
    Sci Signal 1:ra1. 2008
    ..Mice in which chondrocyte-specific deletion of Casr was induced between E16 and E18 were viable but showed delayed growth plate development. Our data show a critical role for the CaSR in early embryogenesis and skeletal development...
  9. ncbi Inactivation of the calcium sensing receptor inhibits E-cadherin-mediated cell-cell adhesion and calcium-induced differentiation in human epidermal keratinocytes
    Chia Ling Tu
    Endocrine Unit, Veteran Affairs Medical Center, University of California, San Francisco, California 94121, USA
    J Biol Chem 283:3519-28. 2008
    ..Our results indicate that the CaR regulates cell survival and Ca(2+)(o)-induced differentiation in keratinocytes at least in part by activating the E-cadherin/PI3K pathway through a Src family tyrosine kinase-mediated signaling...
  10. ncbi The role of the calcium sensing receptor in regulating intracellular calcium handling in human epidermal keratinocytes
    Chia Ling Tu
    Endocrine Unit, Department of Medicine, Veteran Affairs Medical Center and University of California, San Francisco, California 94121, USA
    J Invest Dermatol 127:1074-83. 2007
    ..Our results indicate that CaR regulates keratinocyte differentiation in part by modulating Ca2+(i) stores via interactions with Ca2+ pumps and channels that regulate those stores...
  11. ncbi Constitutive activity of the osteoblast Ca2+-sensing receptor promotes loss of cancellous bone
    Melita M Dvorak
    Endocrine Research Unit, Department of Veterans Affairs Medical Center, 4150 Clement Street, University of California, San Francisco, California 94121, USA
    Endocrinology 148:3156-63. 2007
    ....
  12. ncbi Complex formation with the Type B gamma-aminobutyric acid receptor affects the expression and signal transduction of the extracellular calcium-sensing receptor. Studies with HEK-293 cells and neurons
    Wenhan Chang
    Endocrine Research Unit, Department of Veterans Affairs Medical Center, Department of Medicine, University of California, San Francisco, California 94121, USA
    J Biol Chem 282:25030-40. 2007
    ....
  13. ncbi Type B gamma-aminobutyric acid receptors modulate the function of the extracellular Ca2+-sensing receptor and cell differentiation in murine growth plate chondrocytes
    Zhiqiang Cheng
    Endocrine Research Unit, Department of Veterans Affairs Medical Center, 4150 Clement Street, San Francisco, CA 94121, USA
    Endocrinology 148:4984-92. 2007
    ..These results support the idea that physical interactions between CaRs and GABA-B-R1s modulate the growth and differentiation of GPCs, potentially by altering the function of CaRs...
  14. pmc Naturally-occurring mutation in the calcium-sensing receptor reveals the significance of extracellular domain loop III region for class C G-protein-coupled receptor function
    Qing Dong
    Department of Pediatrics, University of California, San Francisco, San Francisco, California 94143 0434, USA
    J Clin Endocrinol Metab 95:E245-52. 2010
    ..Most mutations are clustered in the N-terminal and Cys-rich regions of the extracellular domain (ECD) and seven-transmembrane domain. Disease-causing mutations are uncommon in the C terminus of ECD...
  15. pmc Negative cross-talk between calcium-sensing receptor and β-catenin signaling systems in colonic epithelium
    Osvaldo Rey
    Unit of Signal Transduction and Gastrointestinal Cancer, Division of Digestive Diseases, Department of Medicine, CURE Digestive Diseases Research Center, David Geffen School of Medicine, UCLA, Los Angeles, California 90095, USA
    J Biol Chem 287:1158-67. 2012
    ..These studies demonstrate that signaling pathways emanating from the CaSR control colonic epithelial cell proliferation in vivo and suggest that the mechanism involves regulation of β-catenin phosphorylation...
  16. pmc Osteoblast extracellular Ca2+ -sensing receptor regulates bone development, mineralization, and turnover
    Melita M Dvorak-Ewell
    Endocrine Research Unit, Department of Veterans Affairs Medical Center, Department of Medicine, University of California, San Francisco, CA 94121, USA
    J Bone Miner Res 26:2935-47. 2011
    ....
  17. pmc IGF-1R signaling in chondrocytes modulates growth plate development by interacting with the PTHrP/Ihh pathway
    Yongmei Wang
    Endocrine Unit, University of California, Veterans Affairs Medical Center, San Francisco, CA, USA
    J Bone Miner Res 26:1437-46. 2011
    ..Our data indicate that the IGF-1R in chondrocytes controls cell growth, survival, and differentiation in embryonic and postnatal GPs in part by suppression of Pthrp expression...
  18. pmc The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and filamin A
    Chia Ling Tu
    Endocrine Unit, Veteran Affairs Medical Center and University of California, San Francisco, California 94121, USA
    J Invest Dermatol 131:1119-28. 2011
    ..These results indicate that Rho is a downstream mediator of CaR in the regulation of Ca(2+)(o)-induced E-cadherin-mediated cell-cell adhesion and keratinocyte differentiation...