{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,4]],"date-time":"2026-05-04T06:23:29Z","timestamp":1777875809645,"version":"3.51.4"},"reference-count":49,"publisher":"SAGE Publications","issue":"7","license":[{"start":{"date-parts":[[2021,7,1]],"date-time":"2021-07-01T00:00:00Z","timestamp":1625097600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Thyroid\u00ae"],"published-print":{"date-parts":[[2021,7]]},"abstract":"<jats:sec>\n                    <jats:title>Background:<\/jats:title>\n                    <jats:p>Loss-of-function mutations of thyrotropin receptor (TSHR) are one of the main causes of congenital hypothyroidism. As for many disease-associated G-protein coupled receptors (GPCRs), these mutations often affect the correct trafficking and maturation of the receptor, thus impairing the expression on the cell surface. Several retained GPCR mutants are able to effectively bind their ligands and to transduce signals when they are forced to the cell surface by degradation inhibition or by treatment with chaperones. Despite the large number of well-characterized retained TSHR mutants, no attempts have been made for rescue. Further, little is known about TSHR degradation pathways. We hypothesize that, similar to other GPCRs, TSHR retained mutants may be at least partially functional if their maturation and membrane expression is facilitated by chaperones or degradation inhibitors.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods:<\/jats:title>\n                    <jats:p>\n                      We performed\n                      <jats:italic toggle=\"yes\">in silico<\/jats:italic>\n                      predictions of the functionality of known TSHR variants and compared the results with available\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      data. Western blot, confocal microscopy, enzyme-linked immunosorbent assays, and dual luciferase assays were used to investigate the effects of degradation pathways inhibition and of chemical chaperone treatments on TSHR variants' maturation and functionality.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results:<\/jats:title>\n                    <jats:p>\n                      We found a high discordance rate between\n                      <jats:italic toggle=\"yes\">in silico<\/jats:italic>\n                      predictions and\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      data for retained TSHR variants, a fact indicative of a conserved potential to initiate signal transduction if these mutants were expressed on the cell surface. We show experimentally that some maturation defective TSHR mutants are able to effectively transduce Gs\/cAMP signaling if their maturation and expression are enhanced by using chemical chaperones. Further, through the characterization of the intracellular retained p.N432D variant, we provide new insights on the TSHR degradation mechanism, as our results suggest that aggregation-prone mutant can be directed toward the autophagosomal pathway instead of the canonical proteasome system.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions:<\/jats:title>\n                    <jats:p>Our study reveals alternative pathways for TSHR degradation. Retained TSHR variants can be functional when expressed on the cell surface membrane, thus opening the possibility of further studies on the pharmacological modulation of TSHR expression and functionality in patients in whom TSHR signaling is disrupted.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1089\/thy.2020.0415","type":"journal-article","created":{"date-parts":[[2021,1,15]],"date-time":"2021-01-15T03:47:02Z","timestamp":1610682422000},"page":"1030-1040","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":0,"title":["Thyrotropin Receptor p.N432D Retained Variant Is Degraded Through an Alternative Lysosomal\/Autophagosomal Pathway and Can Be Functionally Rescued by Chemical Chaperones"],"prefix":"10.1177","volume":"31","author":[{"given":"Elisa Stellaria","family":"Grassi","sequence":"first","affiliation":[{"name":"Department of Medical Biotechnology and Translational Medicine (BIOMETRA), University of 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Italy."},{"name":"Laboratory of Endocrine and Metabolic Research, Istituto Auxologico Italiano IRCCS, Milan, Italy."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luca","family":"Persani","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology and Translational Medicine (BIOMETRA), University of Milan, Milan, Italy."},{"name":"Laboratory of Endocrine and Metabolic Research, Istituto Auxologico Italiano IRCCS, Milan, 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