{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T15:51:55Z","timestamp":1753890715010,"version":"3.41.2"},"reference-count":36,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,4,12]],"date-time":"2021-04-12T00:00:00Z","timestamp":1618185600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Endocrinol."],"abstract":"<jats:sec><jats:title>Purpose<\/jats:title><jats:p>Laron syndrome (LS) is a severe growth disorder caused by<jats:italic>GHR<\/jats:italic>gene mutation or post-receptor pathways defect. The clinical features of these patients collected in our present study were summarized,<jats:italic>GHR<\/jats:italic>gene variants were investigated and further<jats:italic>in vitro<\/jats:italic>functional verification was carried out.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Four patients with LS were collected, their clinical characteristics were summarized, genomic DNA was extracted, and<jats:italic>GHR<\/jats:italic>gene was amplified and sequenced. GHR wild type (GHR-WT) and mutant GHR expression plasmids were constructed, and transiently transfected into HepG2 cells and HEK293T cells to observe the subcellular distribution of the GHR protein by immunofluorescence and to determine the expression of GHR and its post-receptor signaling pathway changes by Western blotting.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>All of the four patients were male, and the median height was -4.72 SDS. Four<jats:italic>GHR<\/jats:italic>gene variants including c.587A&amp;gt;C (p.Y196S), c.766C&amp;gt;T (p.Q256*), c.808A&amp;gt;G (p.I270V) and c.1707-1710del (p.E570Afs*30) were identified, and the latter two were novel mutations. The results of mutant GHR plasmids transfection experiments and immunofluorescence assay showed that the subcellular distribution of GHR-Q256* and GHR-E570Afs*30 mutant proteins in HepG2 and HEK293T cells presented with a unique ring-like pattern, gathering around the nucleus, while GHR-Y196S mutant protein was evenly distributed on HepG2 cell membrane similar to GHR-WT. The GHR protein levels of HepG2 cells transiently transfected with GHR-Y196S, GHR-Q256* and GHR-E570Afs*30 were all significantly lower when compared with cells transfected with GHR-WT (P&amp;lt;0.05). Further mutant GHR post-receptor signal transduction investigation demonstrated that GH induced phosphorylated STAT5 levels of HepG2 cells transfected with three mutant plasmids were all significantly decreased in comparison with that of GHR-WT (P&amp;lt;0.05).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Two novel<jats:italic>GHR<\/jats:italic>gene mutations (I270V and E570Afs*30) were found in our patients with LS. GHR mutations influenced the subcellular distribution and GHR protein levels, then led to the impaired post-receptor signal transduction, suggesting that the<jats:italic>GHR<\/jats:italic>mutations contributed to the pathological condition of LS patients.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fendo.2021.605736","type":"journal-article","created":{"date-parts":[[2021,4,12]],"date-time":"2021-04-12T07:49:00Z","timestamp":1618213740000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Identification and In Vitro Functional Verification of Two Novel Mutations of GHR Gene in the Chinese Children with Laron 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