TY - JOUR
T1 - Impaired iloprost-induced platelet inhibition and phosphoproteome changes in patients with confirmed pseudohypoparathyroidism type Ia, linked to genetic mutations in GNAS
AU - Swieringa, Frauke
AU - Solari, Fiorella A.
AU - Pagel, Oliver
AU - Beck, Florian
AU - Huang, Jingnan
AU - Feijge, Marion A. H.
AU - Jurk, Kerstin
AU - Koerver-Keularts, Irene M. L. W.
AU - Mattheij, Nadine J. A.
AU - Faber, Joerg
AU - Pohlenz, Joachim
AU - Russo, Alexandra
AU - Stumpel, Connie T. R. M.
AU - Schrander, Dirk E.
AU - Zieger, Barbara
AU - van Der Meijden, Paola E. J.
AU - Zahedi, Rene P.
AU - Sickmann, Albert
AU - Heemskerk, Johan W. M.
N1 - Funding Information:
This research was supported by the Ministerium für Innovation, Wissenschaft und Forschung from Nordrhein-Westfalen, the Cardiovascular Centre (HVC) of Maastricht University Medical Centre+, the Centre for Molecular Translational Medicine (INCOAG, MICRO-BAT), the German Federal Ministry of Education and Research (BMBF 01EO1503) and the Deutsche Forschungsgemeinschaft (ZA 639/4-1 and JU 2735/2-1). FS was supported by the Alexander von Humboldt Foundation. This work also received funding from the European Union’s Horizon 2020 research and innovation program under Marie Sklodowska-Curie grant agreement TAPAS No. 766118. JH is enrolled in a joint doctoral program of the universities of Maastricht (The Netherlands) and Santiago de Compostella (Spain).
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/7/9
Y1 - 2020/7/9
N2 - Patients diagnosed with pseudohypoparathyroidism type Ia (PHP Ia) suffer from hormonal resistance and abnormal postural features, in a condition classified as Albright hereditary osteodystrophy (AHO) syndrome. This syndrome is linked to a maternally inherited mutation in the GNAS complex locus, encoding for the GTPase subunit Gs alpha. Here, we investigated how platelet phenotype and omics analysis can assist in the often difficult diagnosis. By coupling to the IP receptor, Gs alpha induces platelet inhibition via adenylyl cyclase and cAMP-dependent protein kinase A (PKA). In platelets from seven patients with suspected AHO, one of the largest cohorts examined, we studied the PKA-induced phenotypic changes. Five patients with a confirmed GNAS mutation, displayed impairments in Gs alpha -dependent VASP phosphorylation, aggregation, and microfluidic thrombus formation. Analysis of the platelet phosphoproteome revealed 2,516 phosphorylation sites, of which 453 were regulated by Gs alpha -PKA. Common changes in the patients were: (1) a joint panel of upregulated and downregulated phosphopeptides; (2) overall PKA dependency of the upregulated phosphopeptides; (3) links to key platelet function pathways. In one patient with GNAS mutation, diagnosed as non-AHO, the changes in platelet phosphoproteome were reversed. This combined approach thus revealed multiple phenotypic and molecular biomarkers to assist in the diagnosis of suspected PHP Ia.
AB - Patients diagnosed with pseudohypoparathyroidism type Ia (PHP Ia) suffer from hormonal resistance and abnormal postural features, in a condition classified as Albright hereditary osteodystrophy (AHO) syndrome. This syndrome is linked to a maternally inherited mutation in the GNAS complex locus, encoding for the GTPase subunit Gs alpha. Here, we investigated how platelet phenotype and omics analysis can assist in the often difficult diagnosis. By coupling to the IP receptor, Gs alpha induces platelet inhibition via adenylyl cyclase and cAMP-dependent protein kinase A (PKA). In platelets from seven patients with suspected AHO, one of the largest cohorts examined, we studied the PKA-induced phenotypic changes. Five patients with a confirmed GNAS mutation, displayed impairments in Gs alpha -dependent VASP phosphorylation, aggregation, and microfluidic thrombus formation. Analysis of the platelet phosphoproteome revealed 2,516 phosphorylation sites, of which 453 were regulated by Gs alpha -PKA. Common changes in the patients were: (1) a joint panel of upregulated and downregulated phosphopeptides; (2) overall PKA dependency of the upregulated phosphopeptides; (3) links to key platelet function pathways. In one patient with GNAS mutation, diagnosed as non-AHO, the changes in platelet phosphoproteome were reversed. This combined approach thus revealed multiple phenotypic and molecular biomarkers to assist in the diagnosis of suspected PHP Ia.
KW - ACTIVATION
KW - COMBINATION
KW - HEREDITARY OSTEODYSTROPHY
KW - PHOSPHORYLATION
KW - PROCOAGULANT ACTIVITY
KW - QUANTIFICATION
KW - REVEALS
KW - ROLES
KW - STIMULATORY G-PROTEIN
KW - THROMBUS FORMATION
U2 - 10.1038/s41598-020-68379-3
DO - 10.1038/s41598-020-68379-3
M3 - Article
C2 - 32647264
SN - 2045-2322
VL - 10
JO - Scientific Reports
JF - Scientific Reports
IS - 1
M1 - 11389
ER -