Induction of Lrp5 HBM-causing mutations in Cathepsin-K expressing cells alters bone metabolism

Kyung Shin Kang, Jung Min Hong, Daniel J. Horan, Kyung Eun Lim, Whitney A. Bullock, Angela Bruzzaniti, Steven Hann, Matthew L. Warman, Alexander Robling

Research output: Contribution to journalArticle

Abstract

High-bone-mass (HBM)-causing missense mutations in the low density lipoprotein receptor-related protein-5 (Lrp5) are associated with increased osteoanabolic action and protection from disuse- and ovariectomy-induced osteopenia. These mutations (e.g., A214V and G171V) confer resistance to endogenous secreted Lrp5/6 inhibitors, such as sclerostin (SOST) and Dickkopf homolog-1 (DKK1). Cells in the osteoblast lineage are responsive to canonical Wnt stimulation, but recent work has indicated that osteoclasts exhibit both indirect and direct responsiveness to canonical Wnt. Whether Lrp5-HBM receptors, expressed in osteoclasts, might alter osteoclast differentiation, activity, and consequent net bone balance in the skeleton, is not known. To address this, we bred mice harboring heterozygous Lrp5 HBM-causing conditional knock-in alleles to Ctsk-Cre transgenic mice and studied the phenotype using DXA, μCT, histomorphometry, serum assays, and primary cell culture. Mice with HBM alleles induced in Ctsk-expressing cells (TG) exhibited higher bone mass and architectural properties compared to non-transgenic (NTG) counterparts. In vivo and in vitro measurements of osteoclast activity, population density, and differentiation yielded significant reductions in osteoclast-related parameters in female but not male TG mice. Droplet digital PCR performed on osteocyte enriched cortical bone tubes from TG and NTG mice revealed that ~8–17% of the osteocyte population (depending on sex) underwent recombination of the conditional Lrp5 allele in the presence of Ctsk-Cre. Further, bone formation parameters in the midshaft femur cortex show a small but significant increase in anabolic action on the endocortical but not periosteal surface. These findings suggest that Wnt/Lrp5 signaling in osteoclasts affects osteoclastogenesis and activity in female mice, but also that some of the changes in bone mass in TG mice might be due to Cre expression in the osteocyte population.

LanguageEnglish (US)
Pages166-175
Number of pages10
JournalBone
Volume120
DOIs
StatePublished - Mar 1 2019

Fingerprint

Low Density Lipoprotein Receptor-Related Protein-5
Cathepsin K
Osteoclasts
Bone and Bones
Mutation
Osteocytes
Alleles
Osteogenesis
Low Density Lipoprotein Receptor-Related Protein-6
Wnt Receptors
Primary Cell Culture
Metabolic Bone Diseases
Ovariectomy
Missense Mutation
Population Density
Osteoblasts
Skeleton
Femur
Transgenic Mice
Genetic Recombination

Keywords

  • Ctsk
  • HBM
  • Lrp5
  • Osteoclasts
  • Resorption

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Physiology
  • Histology

Cite this

Induction of Lrp5 HBM-causing mutations in Cathepsin-K expressing cells alters bone metabolism. / Kang, Kyung Shin; Hong, Jung Min; Horan, Daniel J.; Lim, Kyung Eun; Bullock, Whitney A.; Bruzzaniti, Angela; Hann, Steven; Warman, Matthew L.; Robling, Alexander.

In: Bone, Vol. 120, 01.03.2019, p. 166-175.

Research output: Contribution to journalArticle

Kang, Kyung Shin ; Hong, Jung Min ; Horan, Daniel J. ; Lim, Kyung Eun ; Bullock, Whitney A. ; Bruzzaniti, Angela ; Hann, Steven ; Warman, Matthew L. ; Robling, Alexander. / Induction of Lrp5 HBM-causing mutations in Cathepsin-K expressing cells alters bone metabolism. In: Bone. 2019 ; Vol. 120. pp. 166-175.
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abstract = "High-bone-mass (HBM)-causing missense mutations in the low density lipoprotein receptor-related protein-5 (Lrp5) are associated with increased osteoanabolic action and protection from disuse- and ovariectomy-induced osteopenia. These mutations (e.g., A214V and G171V) confer resistance to endogenous secreted Lrp5/6 inhibitors, such as sclerostin (SOST) and Dickkopf homolog-1 (DKK1). Cells in the osteoblast lineage are responsive to canonical Wnt stimulation, but recent work has indicated that osteoclasts exhibit both indirect and direct responsiveness to canonical Wnt. Whether Lrp5-HBM receptors, expressed in osteoclasts, might alter osteoclast differentiation, activity, and consequent net bone balance in the skeleton, is not known. To address this, we bred mice harboring heterozygous Lrp5 HBM-causing conditional knock-in alleles to Ctsk-Cre transgenic mice and studied the phenotype using DXA, μCT, histomorphometry, serum assays, and primary cell culture. Mice with HBM alleles induced in Ctsk-expressing cells (TG) exhibited higher bone mass and architectural properties compared to non-transgenic (NTG) counterparts. In vivo and in vitro measurements of osteoclast activity, population density, and differentiation yielded significant reductions in osteoclast-related parameters in female but not male TG mice. Droplet digital PCR performed on osteocyte enriched cortical bone tubes from TG and NTG mice revealed that ~8–17{\%} of the osteocyte population (depending on sex) underwent recombination of the conditional Lrp5 allele in the presence of Ctsk-Cre. Further, bone formation parameters in the midshaft femur cortex show a small but significant increase in anabolic action on the endocortical but not periosteal surface. These findings suggest that Wnt/Lrp5 signaling in osteoclasts affects osteoclastogenesis and activity in female mice, but also that some of the changes in bone mass in TG mice might be due to Cre expression in the osteocyte population.",
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