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Calcium-sensing receptor sequencing in 21 patients with idiopathic or familial parathyroid disorder: pitfalls and characterization of a novel I32 V loss-of-function mutation

  • Endocrine Genetics/Epigenetics
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Abstract

The calcium-sensing receptor (CaSR) is a G-protein-coupled receptor with a crucial role in calcium homeostasis. Mutations in the CaSR gene may lead to specific parathyroid disorders due to either gain-of-function (autosomal dominant hypercalciuric hypocalcemia; ADHH) or loss-of-function (familial hypocalciuric hypercalcemia; FHH). Our aim was to evaluate CaSR mutations as a cause of disease in selected patients. We identified and recruited patients with phenotypes suggestive of CaSR-related parathyroid disorders. DNA was extracted, and CaSR gene was sequenced. Live-ratiometric measurements of intracellular [Ca2+] and Western blot assays for evaluation of MAPK phosphorylation in response to changes in extracellular [Ca2+] were performed in transiently transfected HEK-293T cells to functionally characterize mutants. A total of 21 patients were evaluated, seven of them with idiopathic hypoparathyroidism (suspected ADHH) and 14 with hyperparathyroidism (suspected FHH). In the latter group two patients were found to harbor missense mutations: a novel heterozygous I32 V mutation in a female index case and a sporadic known R185Q mutation in a 1-year-old girl. In-vitro functional studies showed that I32 V is an inactivating mutation. In our study, most patients had normal CaSR sequencing. This suggests that phenotypic pitfalls may occur at time of patients’ selection for CaSR sequencing. In one patient with strong positive pre-test probability based on both familial history and appropriate phenotype, a novel I32 V mutation leading to FHH was identified and characterized. In cases of familial parathyroid disorders, CaSR sequencing should be performed, but if negative, one should consider involvement of alternative genes or mechanisms.

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Acknowledgments

The authors would like to thank Prof. Tally Naveh-Many and Prof. Justin Silver for technical assistance, as well as Pr. Benjamin Glaser for support of the study and critical review of the manuscript. The authors also thank Dr. Victoria Terletzky, Dr. Anat Tsur, Dr. Larissa Shustin, Dr. Merav Frankel, Prof. Yair Liel, Dr. Rivka Backenroth-Maayan, Dr Leanna Shkolnik-Tripto, Prof. Iris Vered, and Prof. Yossef Weisman, for cooperation in the collection of clinical and laboratory data. Thanks to Shirley smith for the help in writing the manuscript. This work was supported by the Research Foundation of the Chief Scientist at the Israel Ministry of Health.

Conflict of interest

The authors disclose no competing interest.

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Authors and Affiliations

  1. Endocrinology and Metabolism Service, Department of Internal Medicine, Hadassah-Hebrew University Medical Center, POB 24035, 91240, Jerusalem, Israel

    Auryan Szalat, Boaz Nachmias & Gabriel Munter

  2. The Kuvin Center for the Study of Infectious and Tropical Diseases, Department of Microbiology and Molecular Genetics, Institute for Medical Research-Israel-Canada, Hadassah-Hebrew University Medical Center, Jerusalem, Israel

    Michal Shahar & Haim Rosen

  3. Center for Research, Prevention and Treatment of Atherosclerosis, Hadassah-Hebrew University Medical Center, Jerusalem, Israel

    Shoshana Shpitzen, Ronen Durst & Eran Leitersdorf

  4. Pediatric Endocrinology Unit, Hadassah-Hebrew University Medical Center, Jerusalem, Israel

    David Gillis

  5. Laboratory for Cellular and Molecular Immunology, Department of Internal Medicine, Hadassah-Hebrew University Medical Center, Jerusalem, Israel

    Dror Mevorach

  6. Department of Human Genetics, Hadassah-Hebrew University Medical Center, Jerusalem, Israel

    Vardiella Meiner

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  1. Auryan Szalat
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  2. Michal Shahar
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Szalat, A., Shahar, M., Shpitzen, S. et al. Calcium-sensing receptor sequencing in 21 patients with idiopathic or familial parathyroid disorder: pitfalls and characterization of a novel I32 V loss-of-function mutation. Endocrine 48, 444–453 (2015). https://doi.org/10.1007/s12020-014-0370-3

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