Mutant Prolactin Receptor and Familial Hyperprolactinemia

Newey, Paul; Gorvin, Caroline M

doi:10.1056/nejmoa1307557

Cited by 112 publications

(110 citation statements)

References 25 publications

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“…WT and mutant pBI-CMV2-GNA11 expression constructs were generated as described (11). Site-directed mutagenesis was used to generate the mutant GNA11 construct using the Quikchange Lightning Site-directed Mutagenesis kit (Agilent Technologies) and gene-specific primers (Sigma-Aldrich), as described (34). This bidirectional pBI-CMV2 vector allows for coexpression of Gα 11 and GFP at equivalent levels (11).…”

Section: Methodsmentioning

confidence: 99%

Knockin mouse with mutant Gα11 mimics human inherited hypocalcemia and is rescued by pharmacologic inhibitors

Roszko

Gorvin

et al. 2017

JCI Insight

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Section: Methodsmentioning

confidence: 99%

Knockin mouse with mutant Gα11 mimics human inherited hypocalcemia and is rescued by pharmacologic inhibitors

Roszko

Gorvin

et al. 2017

JCI Insight

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“…The bidirectional pBI‐CMV2 cloning vector was used because it facilitated the co‐expression of Gα 11 and GFP,11, 12 and site‐directed mutagenesis was used to generate the mutant GNA11 construct using the Quikchange Lightning Site‐directed Mutagenesis kit (Agilent Technologies, Santa Clara, CA, USA) and gene‐specific primers (Sigma‐Aldrich), as described 19. Cells were maintained in DMEM‐Glutamax media (Thermo‐Fisher, Waltham, MA, USA) with 10% fetal bovine serum (Gibco, Thermo‐Fisher) and 400 μg/mL geneticin (Thermo‐Fisher) at 37°C, 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

A G-protein Subunit-α11 Loss-of-Function Mutation, Thr54Met, Causes Familial Hypocalciuric Hypercalcemia Type 2 (FHH2)

Gorvin

Cranston

Hannan

et al. 2016

Journal of Bone and Mineral Research

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Familial hypocalciuric hypercalcemia (FHH) is a genetically heterogeneous disorder with three variants, FHH1 to FHH3. FHH1 is caused by loss‐of‐function mutations of the calcium‐sensing receptor (CaSR), a G‐protein coupled receptor that predominantly signals via G‐protein subunit alpha‐11 (Gα11) to regulate calcium homeostasis. FHH2 is the result of loss‐of‐function mutations in Gα11, encoded by GNA11, and to date only two FHH2‐associated Gα11 missense mutations (Leu135Gln and Ile200del) have been reported. FHH3 is the result of loss‐of‐function mutations of the adaptor protein‐2 σ‐subunit (AP2σ), which plays a pivotal role in clathrin‐mediated endocytosis. We describe a 65‐year‐old woman who had hypercalcemia with normal circulating parathyroid hormone concentrations and hypocalciuria, features consistent with FHH, but she did not have CaSR and AP2σ mutations. Mutational analysis of the GNA11 gene was therefore undertaken, using leucocyte DNA, and this identified a novel heterozygous GNA11 mutation (c.161C>T; p.Thr54Met). The effect of the Gα11 variant was assessed by homology modeling of the related Gαq protein and by measuring the CaSR‐mediated intracellular calcium (Ca2+ i) responses of HEK293 cells, stably expressing CaSR, to alterations in extracellular calcium (Ca2+ o) using flow cytometry. Three‐dimensional modeling revealed the Thr54Met mutation to be located at the interface between the Gα11 helical and GTPase domains, and to likely impair GDP binding and interdomain interactions. Expression of wild‐type and the mutant Gα11 in HEK293 cells stably expressing CaSR demonstrate that the Ca2+ i responses after stimulation with Ca2+ o of the mutant Met54 Gα11 led to a rightward shift of the concentration‐response curve with a significantly (p < 0.01) increased mean half‐maximal concentration (EC50) value of 3.88 mM (95% confidence interval [CI] 3.76–4.01 mM), when compared with the wild‐type EC50 of 2.94 mM (95% CI 2.81–3.07 mM) consistent with a loss‐of‐function. Thus, our studies have identified a third Gα11 mutation (Thr54Met) causing FHH2 and reveal a critical role for the Gα11 interdomain interface in CaSR signaling and Ca2+ o homeostasis. © 2016 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research (ASBMR).

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“…We therefore suspected that a germline PRLR mutation might also be a specific genetic cause of prolactinoma in humans. This suspicion was reinforced in late 2013 by the identification of the first inactivating germline mutation of the human PRLR gene in 3 sisters with familial idiopathic hyperprolactinemia [12]. The diagnosis was based on high plasma PRL levels in the absence of medications or medical conditions known to provoke hyperprolactinemia.…”

Section: Introductionmentioning

confidence: 99%

Germline Prolactin Receptor Mutation Is Not a Major Cause of Sporadic Prolactinoma in Humans

et al. 2015

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Background/Aims: No genetic anomalies specifically predisposing humans to prolactinomas have so far been identified. The prolactin receptor (PRLR) is a good candidate, however, as Prlr knockout mice develop prolactinomas, and a case of familial hyperprolactinemia has been linked to PRLR mutation. The main objective of this study was to detect germline PRLR mutations in patients with sporadic prolactinomas unrelated to AIP or MEN1 mutation. Methods: We sequenced all PRLR exons and intron-exon junctions on genomic DNA from 88 patients with a median age of 24 years. Results: We identified 4 PRLR variations (p.Ile76Val, p.Ile146Leu, p.Glu108Lys and p.Glu554Gln) in 16 patients. One patient had the rare variant p.Glu554Gln in the heterozygous state. Another patient had the extremely rare p.Glu108Lys variant described here for the first time. The other 2 variants (p.Ile76Val and p.Ile146Leu) are relatively common in the general population. All these 4 variants have been functionally tested in vitro and have no effect on PRLR expression, localization and signaling after prolactin stimulation. Conclusion: Inactivating germline variations of PRLR are not associated with sporadic prolactinoma in this series. Nevertheless, somatic disruption of PRLR has not been excluded in this subset of pituitary tumors.

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Mutant Prolactin Receptor and Familial Hyperprolactinemia

Cited by 112 publications

References 25 publications

Knockin mouse with mutant Gα11 mimics human inherited hypocalcemia and is rescued by pharmacologic inhibitors

Knockin mouse with mutant Gα11 mimics human inherited hypocalcemia and is rescued by pharmacologic inhibitors

A G-protein Subunit-α11 Loss-of-Function Mutation, Thr54Met, Causes Familial Hypocalciuric Hypercalcemia Type 2 (FHH2)

Germline Prolactin Receptor Mutation Is Not a Major Cause of Sporadic Prolactinoma in Humans

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