{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,28]],"date-time":"2026-03-28T07:02:00Z","timestamp":1774681320991,"version":"3.50.1"},"reference-count":15,"publisher":"Oxford University Press (OUP)","issue":"11","funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000051","name":"National Human Genome Research Institute","doi-asserted-by":"publisher","award":["HG200328\u201319"],"award-info":[{"award-number":["HG200328\u201319"]}],"id":[{"id":"10.13039\/100000051","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000051","name":"National Human Genome Research Institute","doi-asserted-by":"publisher","award":["HG200359\u201316"],"award-info":[{"award-number":["HG200359\u201316"]}],"id":[{"id":"10.13039\/100000051","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000069","name":"National Institute of Arthritis and Musculoskeletal and Skin Diseases","doi-asserted-by":"publisher","award":["AR082209\u201303"],"award-info":[{"award-number":["AR082209\u201303"]}],"id":[{"id":"10.13039\/100000069","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,5,17]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Purpose<\/jats:title>\n                    <jats:p>Identifying individuals with pathogenic variants for RYR1-related Malignant Hyperthermia Susceptibility (MHS) could reduce morbidity and mortality due to MH reactions. Realization of this goal requires knowledge of variant pathogenicity, and proper weighting of functional assays is important for accurate variant classification. Caffeine-induced Ca2+ release assays can be used to support pathogenicity per the ClinGen Variant Curation Expert Panel (VCEP). However, the caffeine-induced Ca2+ release assay lacks formal validation with known pathogenic and benign variants.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Fifteen benign\/likely benign and six pathogenic\/likely pathogenic RYR1 variants were used to calibrate the caffeine-induced Ca2+ release assay using a multi-mode microplate reader. Five variants of unknown significance (VUS) were assayed for possible reclassification.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Our data support use of the caffeine-induced Ca2+ release assay at a moderate weight per the American College of Medical Genetics and Genomics pathogenicity criteria schema with a positive likelihood ratio of 12.14:1 (pathogenicity) and a negative likelihood ratio of 0.22:1 (4.5:1 benignity). Using this validated assay, two of five VUS were reclassified as likely benign.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Formal validation of the caffeine-induced Ca2+ release assay supports the VCEP functional criteria weighting at moderate strength based on these data. Additional variants should be assayed to shift more from VUS to benign or pathogenic classifications.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1093\/hmg\/ddaf038","type":"journal-article","created":{"date-parts":[[2025,4,7]],"date-time":"2025-04-07T12:42:32Z","timestamp":1744029752000},"page":"945-951","source":"Crossref","is-referenced-by-count":0,"title":["Calibrating a functional assay for variant classification in\n                    <i>RYR1<\/i>\n                    -related malignant hyperthermia susceptibility"],"prefix":"10.1093","volume":"34","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-4629-9826","authenticated-orcid":false,"given":"Evan Z","family":"Ying","sequence":"first","affiliation":[{"name":"Center for Precision Health Research, National Human Genome Research Institute, National Institutes of Health , 50 South Drive, Bethesda, MD 20892 ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amber","family":"Douglass","sequence":"additional","affiliation":[{"name":"Center for Precision Health Research, National Human Genome Research Institute, National Institutes of Health , 50 South Drive, Bethesda, MD 20892 ,","place":["United States"]},{"name":"Department of Microbiology Immunology and Tropical Medicine, The George Washington University , 2300 I St. NW, Washington, DC 20037 ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marwan A","family":"Hawari","sequence":"additional","affiliation":[{"name":"Center for Precision Health Research, National Human Genome Research Institute, National Institutes of Health , 50 South Drive, Bethesda, MD 20892 ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Linda","family":"Groom","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Physiology, University of Rochester Medical Center , 601 Elmwood Avenue, Rochester, NY 14642 ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kathryn","family":"Stowell","sequence":"additional","affiliation":[{"name":"School of Food Technology and Natural Sciences, Massey University , Tennent Drive, Palmerston North 4410 ,","place":["New Zealand"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert T","family":"Dirksen","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Physiology, University of Rochester Medical Center , 601 Elmwood Avenue, Rochester, NY 14642 ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1012-6358","authenticated-orcid":false,"given":"Jennifer J","family":"Johnston","sequence":"additional","affiliation":[{"name":"Center for Precision Health Research, National Human Genome Research Institute, National Institutes of Health , 50 South Drive, Bethesda, MD 20892 ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0197-3811","authenticated-orcid":false,"given":"Leslie G","family":"Biesecker","sequence":"additional","affiliation":[{"name":"Center for Precision Health Research, National Human Genome Research Institute, National Institutes of Health , 50 South Drive, Bethesda, MD 20892 ,","place":["United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2025,4,7]]},"reference":[{"key":"2026032801425047700_ref1","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1186\/s13023-015-0310-1","article-title":"Malignant hyperthermia: a review","volume":"10","author":"Rosenberg","year":"2015","journal-title":"Orphanet J Rare Dis"},{"key":"2026032801425047700_ref2","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1111\/j.1749-6632.1993.tb38060.x","article-title":"The role of the calcium release channel of skeletal muscle sarcoplasmic reticulum in malignant hyperthermia","volume":"707","author":"MacLennan","year":"1993","journal-title":"Ann N Y Acad Sci"},{"key":"2026032801425047700_ref3","doi-asserted-by":"crossref","first-page":"944","DOI":"10.1016\/j.bja.2018.06.028","article-title":"Genetic epidemiology of malignant hyperthermia in the UK","volume":"121","author":"Miller","year":"2018","journal-title":"Br J Anaesth"},{"key":"2026032801425047700_ref4","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1093\/bja\/aev225","article-title":"European malignant hyperthermia group guidelines for investigation of malignant hyperthermia susceptibility","volume":"115","author":"Hopkins","year":"2015","journal-title":"Br J Anaesth"},{"key":"2026032801425047700_ref5","doi-asserted-by":"crossref","first-page":"4087","DOI":"10.1093\/hmg\/ddac145","article-title":"Updated variant curation expert panel criteria and pathogenicity classifications for 251 variants for RYR1-related malignant hyperthermia susceptibility","volume":"31","author":"Johnston","year":"2022","journal-title":"Hum Mol Genet"},{"key":"2026032801425047700_ref6","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1038\/gim.2015.30","article-title":"Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology","volume":"17","author":"Richards","year":"2015","journal-title":"Genet Med"},{"key":"2026032801425047700_ref7","doi-asserted-by":"crossref","first-page":"26332","DOI":"10.1074\/jbc.272.42.26332","article-title":"Caffeine and halothane sensitivity of intracellular Ca2+ release is altered by 15 calcium release channel (ryanodine receptor) mutations associated with malignant hyperthermia and\/or central core disease","volume":"272","author":"Tong","year":"1997","journal-title":"J Biol Chem"},{"key":"2026032801425047700_ref8","doi-asserted-by":"crossref","first-page":"101228","DOI":"10.1016\/j.gim.2024.101228","article-title":"The clinical genome resource (ClinGen): advancing genomic knowledge through global curation","volume":"27","author":"ClinGen Consortium","year":"2025","journal-title":"Genet Med"},{"key":"2026032801425047700_ref9","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1186\/s13073-019-0690-2","article-title":"Recommendations for application of the functional evidence PS3\/BS3 criterion using the ACMG\/AMP sequence variant interpretation framework","volume":"12","author":"Brnich","year":"2019","journal-title":"Genome Med"},{"key":"2026032801425047700_ref10","doi-asserted-by":"crossref","first-page":"1288","DOI":"10.1038\/s41436-021-01125-w","article-title":"Variant curation expert panel recommendations for RYR1 pathogenicity classifications in malignant hyperthermia susceptibility","volume":"23","author":"Johnston","year":"2021","journal-title":"Genet Med."},{"key":"2026032801425047700_ref11","doi-asserted-by":"crossref","first-page":"948","DOI":"10.1016\/j.bbamcr.2010.11.009","article-title":"Mechanistic models for muscle diseases and disorders originating in the sarcoplasmic reticulum","volume":"1813","author":"MacLennan","year":"2011","journal-title":"Biochim Biophys Acta"},{"key":"2026032801425047700_ref12","doi-asserted-by":"crossref","first-page":"7947","DOI":"10.1038\/ncomms8947","article-title":"Crystal structures of ryanodine receptor SPRY1 and tandem-repeat domains reveal a critical FKBP12 binding determinant","volume":"6","author":"Yuchi","year":"2015","journal-title":"Nat Commun"},{"key":"2026032801425047700_ref13","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.1038\/gim.2017.210","article-title":"Modeling the ACMG\/AMP variant classification guidelines as a Bayesian classification framework","volume":"20","author":"Tavtigian","year":"2018","journal-title":"Genet Med"},{"key":"2026032801425047700_ref14","doi-asserted-by":"crossref","first-page":"1734","DOI":"10.1002\/humu.24088","article-title":"Fitting a naturally scaled point system to the ACMG\/AMP variant classification guidelines","volume":"41","author":"Tavtigian","year":"2020","journal-title":"Hum Mutat"},{"key":"2026032801425047700_ref15","doi-asserted-by":"crossref","first-page":"879","DOI":"10.1016\/j.bja.2022.08.029","article-title":"Functional analysis of RYR1 variants in patients with confirmed susceptibility to malignant hyperthermia","volume":"129","author":"White","year":"2022","journal-title":"Br J Anaesth"}],"container-title":["Human Molecular Genetics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/hmg\/article-pdf\/34\/11\/945\/62883457\/ddaf038.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/hmg\/article-pdf\/34\/11\/945\/62883457\/ddaf038.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,28]],"date-time":"2026-03-28T05:43:01Z","timestamp":1774676581000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/hmg\/article\/34\/11\/945\/8107667"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,7]]},"references-count":15,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2025,5,17]]}},"URL":"https:\/\/doi.org\/10.1093\/hmg\/ddaf038","relation":{},"ISSN":["0964-6906","1460-2083"],"issn-type":[{"value":"0964-6906","type":"print"},{"value":"1460-2083","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2025,6,1]]},"published":{"date-parts":[[2025,4,7]]}}}