Dephosphorylation of the NPR2 guanylyl cyclase contributes to inhibition of bone growth by fibroblast growth factor

Shuhaibar, Leia C.; Robinson, Jerid W.; Vigone, Giulia; Shuhaibar, Ninna; Egbert, Jeremy R.; Baena, Valentina; Uliasz, Tracy F.; Kaback, Deborah; Yee, Siu Pok; Feil, Robert; Fisher, Melanie C.; Dealy, Caroline N.; Potter, Lincoln R.; Jaffe, Laurinda A.

doi:10.7554/elife.31343

Cited by 28 publications

(31 citation statements)

References 68 publications

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“…The latter might be linked to other signaling systems of which the cytoplasmic enzymes that control the phosphorylation of Npr2 are currently unknown (Schmidt and others 2018). More recently, such a link between FGFR3 signaling and the phosphorylation of Npr2 has been detected in chondrocytes (Robinson and others 2017; Shuhaibar and others 2017).…”

Section: Phosphorylation Of Npr2 and Binding Of The Ligand Cnp Are Bomentioning

confidence: 92%

Sensory Neurons: The Formation of T-Shaped Branches Is Dependent on a cGMP-Dependent Signaling Cascade

2020

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Axon bifurcation – a specific form of branching of somatosensory axons characterized by the splitting of the growth cone – is mediated by a cGMP-dependent signaling cascade composed of the extracellular ligand CNP (C-type natriuretic peptide), the transmembrane receptor guanylyl cyclase Npr2 (natriuretic peptide receptor 2), and the kinase cGKI (cGMP-dependent protein kinase I). In the absence of any one of these components, the formation of T-shaped axonal branches is impaired in neurons from DRGs (dorsal root ganglia), CSGs (cranial sensory ganglia) and MTNs (mesencephalic trigeminal neurons) in the murine spinal cord or hindbrain. Instead, axons from DRGs or from CSGs extend only either in an ascending or descending direction, while axons from MTNs either elongate within the hindbrain or extend via the trigeminal ganglion to the masseter muscles. Collateral formation from non-bifurcating stem axons is not affected by impaired cGMP signaling. Activation of Npr2 requires both binding of the ligand CNP as well as phosphorylation of serine and threonine residues at the juxtamembrane regions of the receptor. The absence of bifurcation results in an altered shape of termination fields of sensory afferents in the spinal cord and resulted in impaired noxious heat sensation and nociception whereas motor coordination appeared normal.

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Section: Phosphorylation Of Npr2 and Binding Of The Ligand Cnp Are Bomentioning

confidence: 92%

Sensory Neurons: The Formation of T-Shaped Branches Is Dependent on a cGMP-Dependent Signaling Cascade

2020

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“…Although the effects of FGFR3-G380R and GC-B-7E expression on naso-anal length of mixed male and female populations have been described (10,24), the effects of sex have not been reported for either murine receptor. Using repeated measures analysis from 4 weeks to 16 weeks of age, we observed that expression of one allele of FGFR3-G380R decreased skeletal length in male mice by 4.7% but only decreased skeletal length by 1.3% in female mice compared to WT controls (Fig.…”

Section: Fgfr3-g380r and Gc-b-7e Exhibit Sex-specific Gene Dosage Effmentioning

confidence: 99%

“…Phosphorylation is absolutely required for CNP-dependent activation of GC-B and alanine substitutions for the known phosphorylation sites yields an enzyme that is not activated by CNP (15)(16)(17). In contrast, conversion of the phosphorylated serines and threonines to glutamate to constitutively mimic the negative charge of the phosphorylated residues produces an enzyme that is activated by CNP but not inactivated by dephosphorylation (20)(21)(22)(23)(24). GC-B knock out mice or GC-B mice with alanine substitutions for the phosphorylated serines and threonines are dwarfed and have smaller growth plates that are similar to those of ACH mice (17,25,26).…”

Section: Introductionmentioning

confidence: 99%

Prevention of guanylyl cyclase–B dephosphorylation rescues achondroplastic dwarfism

Wagner¹,

Robinson²,

Lin³

et al. 2021

JCI Insight

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Activating mutations in the fibroblast growth factor receptor 3 (FGFR3) or inactivating mutations in guanylyl cyclase B (GC-B), also known as NPR-B or Npr2, cause short-limbed dwarfism. FGFR3 activation causes dephosphorylation and inactivation of GC-B, but the contribution of GC-B dephosphorylation to achondroplasia (ACH) is unknown. GC-B 7E/7E mice that express a glutamatesubstituted version of GC-B that cannot be inactivated by dephosphorylation were bred with mice expressing FGFR3-G380R, the most common human ACH mutation, to determine if GC-B dephosphorylation is required for ACH. Crossing GC-B 7E/7E mice with FGFR3 G380R/G380R mice increased naso-anal and long (tibia and femur), but not cranial, bone length twice as much as crossing GC-B 7E/7E mice with FGFR3 WT/WT mice from 4 to 16 weeks of age. Consistent with increased GC-B activity rescuing ACH, long bones from the GC-B 7E/7E /FGFR3 G380R/G380R mice were not shorter than those from GC-B WT/WT /FGFR3 WT/WT mice. At two weeks of age, male but not female FGFR3 G380R/G380R mice had shorter long bones and smaller growth plate hypertrophic zones, whereas female but not male GC-B 7E/7E mice had longer bones and larger hypertrophic zones. In two-week old males, crossing FGFR3 G380R/G380R mice with GC-B 7E/7E mice increased long bone length and hypertrophic zone area to levels observed in mice expressing wild type versions of both receptors. We conclude that preventing GC-B dephosphorylation rescues reduced axial and appendicular skeleton growth in a mouse model of achondroplasia.

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“…LB-100 counteracts the inactivation of Npr2 by FGF in growth plate chondrocytes. NPR2 activity in chondrocytes of intact growth plates was measured as previously described, using mice expressing a FRET sensor for cGMP, cGi500 (27). The mice were WT for Fgfr3.…”

Section: Resultsmentioning

confidence: 99%

Phosphatase inhibition by LB-100 enhances BMN-111 stimulation of bone growth

Shuhaibar¹,

Kaci²,

Egbert³

et al. 2021

JCI Insight

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Activating mutations in fibroblast growth factor receptor 3 (FGFR3) and inactivating mutations in the natriuretic peptide receptor 2 (NPR2) guanylyl cyclase both result in decreased production of cyclic GMP in chondrocytes and severe short stature, causing achondroplasia (ACH) and acromesomelic dysplasia, type Maroteaux, respectively. Previously, we showed that an NPR2 agonist BMN-111 (vosoritide) increases bone growth in mice mimicking ACH ( Fgfr3 Y367C/+ ). Here, because FGFR3 signaling decreases NPR2 activity by dephosphorylating the NPR2 protein, we tested whether a phosphatase inhibitor (LB-100) could enhance BMN-111–stimulated bone growth in ACH. Measurements of cGMP production in chondrocytes of living tibias, and of NPR2 phosphorylation in primary chondrocytes, showed that LB-100 counteracted FGF-induced dephosphorylation and inactivation of NPR2. In ex vivo experiments with Fgfr3 Y367C/+ mice, the combination of BMN-111 and LB-100 increased bone length and cartilage area, restored chondrocyte terminal differentiation, and increased the proliferative growth plate area, more than BMN-111 alone. The combination treatment also reduced the abnormal elevation of MAP kinase activity in the growth plate of Fgfr3 Y367C/+ mice and improved the skull base anomalies. Our results provide a proof of concept that a phosphatase inhibitor could be used together with an NPR2 agonist to enhance cGMP production as a therapy for ACH.

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Dephosphorylation of the NPR2 guanylyl cyclase contributes to inhibition of bone growth by fibroblast growth factor

Cited by 28 publications

References 68 publications

Sensory Neurons: The Formation of T-Shaped Branches Is Dependent on a cGMP-Dependent Signaling Cascade

Sensory Neurons: The Formation of T-Shaped Branches Is Dependent on a cGMP-Dependent Signaling Cascade

Prevention of guanylyl cyclase–B dephosphorylation rescues achondroplastic dwarfism

Phosphatase inhibition by LB-100 enhances BMN-111 stimulation of bone growth

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