Divergent effects of strontium and calcium-sensing receptor positive allosteric modulators (calcimimetics) on human osteoclast activity

Natalie A Diepenhorst, Katie Leach, Andrew N Keller, Patricia Rueda, Anna E Cook, Tracie L Pierce, Cameron Nowell, Philippe Pastoureau, Massimo Sabatini, Roger J Summers, William N Charman, Patrick M Sexton, Arthur Christopoulos, Christopher J Langmead

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Background and Purpose: Strontium ranelate, a drug approved and until recently used for the treatment of osteoporosis, mediates its effects on bone at least in part via the calcium-sensing (CaS) receptor. However, it is not known whether bone-targeted CaS receptor positive allosteric modulators (PAMs; calcimimetics) represent an alternative (or adjunctive) therapy to strontium (Sr2+ o). Experimental Approach: We assessed three structurally distinct calcimimetics [cinacalcet, AC-265347 and a benzothiazole tri-substituted urea (BTU-compound 13)], alone and in combination with extracellular calcium (Ca2+ o) or Sr2+ o, in G protein-dependent signalling assays and trafficking experiments in HEK293 cells and their effects on cell differentiation, tartrate-resistant acid phosphatase (TRAP) activity and hydroxyapatite resorption assays in human blood-derived osteoclasts. Key Results: Sr2+ o activated CaS receptor-dependent signalling in HEK293 cells in a similar manner to Ca2+ o, and inhibited the maturation, TRAP expression and hydroxyapatite resorption capacity of human osteoclasts. Calcimimetics potentiated Ca2+ o- and Sr2+ o-mediated CaS receptor signalling in HEK293 cells with distinct biased profiles, and only cinacalcet chaperoned an endoplasmic reticulum-retained CaS mutant receptor to the cell surface in HEK293 cells, indicative of a conformational state different from that engendered by AC-265347 and BTU-compound 13. Intriguingly, only cinacalcet modulated human osteoclast function, reducing TRAP activity and profoundly inhibiting resorption. Conclusion and Implications: Although AC-265347 and BTU-compound 13 potentiated Ca2+ o- and Sr2+ o-induced CaS receptor activation, they neither replicated nor potentiated the ability of Sr2+ o to inhibit human osteoclast function. In contrast, the FDA-approved calcimimetic, cinacalcet, inhibited osteoclast TRAP activity and hydroxyapatite resorption, which may contribute to its clinical effects on bone mineral density. Linked Articles: This article is part of a themed section on Molecular Pharmacology of GPCRs. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.21/issuetoc.

Original languageEnglish
Pages (from-to)4095-4108
Number of pages14
JournalBritish Journal of Pharmacology
Volume175
Issue number21
DOIs
Publication statusPublished - 1 Nov 2018

Cite this

@article{bae5b541c7a54c3885eaf315bcb4e91d,
title = "Divergent effects of strontium and calcium-sensing receptor positive allosteric modulators (calcimimetics) on human osteoclast activity",
abstract = "Background and Purpose: Strontium ranelate, a drug approved and until recently used for the treatment of osteoporosis, mediates its effects on bone at least in part via the calcium-sensing (CaS) receptor. However, it is not known whether bone-targeted CaS receptor positive allosteric modulators (PAMs; calcimimetics) represent an alternative (or adjunctive) therapy to strontium (Sr2+ o). Experimental Approach: We assessed three structurally distinct calcimimetics [cinacalcet, AC-265347 and a benzothiazole tri-substituted urea (BTU-compound 13)], alone and in combination with extracellular calcium (Ca2+ o) or Sr2+ o, in G protein-dependent signalling assays and trafficking experiments in HEK293 cells and their effects on cell differentiation, tartrate-resistant acid phosphatase (TRAP) activity and hydroxyapatite resorption assays in human blood-derived osteoclasts. Key Results: Sr2+ o activated CaS receptor-dependent signalling in HEK293 cells in a similar manner to Ca2+ o, and inhibited the maturation, TRAP expression and hydroxyapatite resorption capacity of human osteoclasts. Calcimimetics potentiated Ca2+ o- and Sr2+ o-mediated CaS receptor signalling in HEK293 cells with distinct biased profiles, and only cinacalcet chaperoned an endoplasmic reticulum-retained CaS mutant receptor to the cell surface in HEK293 cells, indicative of a conformational state different from that engendered by AC-265347 and BTU-compound 13. Intriguingly, only cinacalcet modulated human osteoclast function, reducing TRAP activity and profoundly inhibiting resorption. Conclusion and Implications: Although AC-265347 and BTU-compound 13 potentiated Ca2+ o- and Sr2+ o-induced CaS receptor activation, they neither replicated nor potentiated the ability of Sr2+ o to inhibit human osteoclast function. In contrast, the FDA-approved calcimimetic, cinacalcet, inhibited osteoclast TRAP activity and hydroxyapatite resorption, which may contribute to its clinical effects on bone mineral density. Linked Articles: This article is part of a themed section on Molecular Pharmacology of GPCRs. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.21/issuetoc.",
author = "Diepenhorst, {Natalie A} and Katie Leach and Keller, {Andrew N} and Patricia Rueda and Cook, {Anna E} and Pierce, {Tracie L} and Cameron Nowell and Philippe Pastoureau and Massimo Sabatini and Summers, {Roger J} and Charman, {William N} and Sexton, {Patrick M} and Arthur Christopoulos and Langmead, {Christopher J}",
year = "2018",
month = "11",
day = "1",
doi = "10.1111/bph.14344",
language = "English",
volume = "175",
pages = "4095--4108",
journal = "British Journal of Pharmacology",
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Divergent effects of strontium and calcium-sensing receptor positive allosteric modulators (calcimimetics) on human osteoclast activity. / Diepenhorst, Natalie A; Leach, Katie; Keller, Andrew N; Rueda, Patricia; Cook, Anna E; Pierce, Tracie L; Nowell, Cameron; Pastoureau, Philippe; Sabatini, Massimo; Summers, Roger J; Charman, William N; Sexton, Patrick M; Christopoulos, Arthur; Langmead, Christopher J.

In: British Journal of Pharmacology, Vol. 175, No. 21, 01.11.2018, p. 4095-4108.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Divergent effects of strontium and calcium-sensing receptor positive allosteric modulators (calcimimetics) on human osteoclast activity

AU - Diepenhorst, Natalie A

AU - Leach, Katie

AU - Keller, Andrew N

AU - Rueda, Patricia

AU - Cook, Anna E

AU - Pierce, Tracie L

AU - Nowell, Cameron

AU - Pastoureau, Philippe

AU - Sabatini, Massimo

AU - Summers, Roger J

AU - Charman, William N

AU - Sexton, Patrick M

AU - Christopoulos, Arthur

AU - Langmead, Christopher J

PY - 2018/11/1

Y1 - 2018/11/1

N2 - Background and Purpose: Strontium ranelate, a drug approved and until recently used for the treatment of osteoporosis, mediates its effects on bone at least in part via the calcium-sensing (CaS) receptor. However, it is not known whether bone-targeted CaS receptor positive allosteric modulators (PAMs; calcimimetics) represent an alternative (or adjunctive) therapy to strontium (Sr2+ o). Experimental Approach: We assessed three structurally distinct calcimimetics [cinacalcet, AC-265347 and a benzothiazole tri-substituted urea (BTU-compound 13)], alone and in combination with extracellular calcium (Ca2+ o) or Sr2+ o, in G protein-dependent signalling assays and trafficking experiments in HEK293 cells and their effects on cell differentiation, tartrate-resistant acid phosphatase (TRAP) activity and hydroxyapatite resorption assays in human blood-derived osteoclasts. Key Results: Sr2+ o activated CaS receptor-dependent signalling in HEK293 cells in a similar manner to Ca2+ o, and inhibited the maturation, TRAP expression and hydroxyapatite resorption capacity of human osteoclasts. Calcimimetics potentiated Ca2+ o- and Sr2+ o-mediated CaS receptor signalling in HEK293 cells with distinct biased profiles, and only cinacalcet chaperoned an endoplasmic reticulum-retained CaS mutant receptor to the cell surface in HEK293 cells, indicative of a conformational state different from that engendered by AC-265347 and BTU-compound 13. Intriguingly, only cinacalcet modulated human osteoclast function, reducing TRAP activity and profoundly inhibiting resorption. Conclusion and Implications: Although AC-265347 and BTU-compound 13 potentiated Ca2+ o- and Sr2+ o-induced CaS receptor activation, they neither replicated nor potentiated the ability of Sr2+ o to inhibit human osteoclast function. In contrast, the FDA-approved calcimimetic, cinacalcet, inhibited osteoclast TRAP activity and hydroxyapatite resorption, which may contribute to its clinical effects on bone mineral density. Linked Articles: This article is part of a themed section on Molecular Pharmacology of GPCRs. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.21/issuetoc.

AB - Background and Purpose: Strontium ranelate, a drug approved and until recently used for the treatment of osteoporosis, mediates its effects on bone at least in part via the calcium-sensing (CaS) receptor. However, it is not known whether bone-targeted CaS receptor positive allosteric modulators (PAMs; calcimimetics) represent an alternative (or adjunctive) therapy to strontium (Sr2+ o). Experimental Approach: We assessed three structurally distinct calcimimetics [cinacalcet, AC-265347 and a benzothiazole tri-substituted urea (BTU-compound 13)], alone and in combination with extracellular calcium (Ca2+ o) or Sr2+ o, in G protein-dependent signalling assays and trafficking experiments in HEK293 cells and their effects on cell differentiation, tartrate-resistant acid phosphatase (TRAP) activity and hydroxyapatite resorption assays in human blood-derived osteoclasts. Key Results: Sr2+ o activated CaS receptor-dependent signalling in HEK293 cells in a similar manner to Ca2+ o, and inhibited the maturation, TRAP expression and hydroxyapatite resorption capacity of human osteoclasts. Calcimimetics potentiated Ca2+ o- and Sr2+ o-mediated CaS receptor signalling in HEK293 cells with distinct biased profiles, and only cinacalcet chaperoned an endoplasmic reticulum-retained CaS mutant receptor to the cell surface in HEK293 cells, indicative of a conformational state different from that engendered by AC-265347 and BTU-compound 13. Intriguingly, only cinacalcet modulated human osteoclast function, reducing TRAP activity and profoundly inhibiting resorption. Conclusion and Implications: Although AC-265347 and BTU-compound 13 potentiated Ca2+ o- and Sr2+ o-induced CaS receptor activation, they neither replicated nor potentiated the ability of Sr2+ o to inhibit human osteoclast function. In contrast, the FDA-approved calcimimetic, cinacalcet, inhibited osteoclast TRAP activity and hydroxyapatite resorption, which may contribute to its clinical effects on bone mineral density. Linked Articles: This article is part of a themed section on Molecular Pharmacology of GPCRs. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.21/issuetoc.

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EP - 4108

JO - British Journal of Pharmacology

JF - British Journal of Pharmacology

SN - 1476-5381

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