{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,8,21]],"date-time":"2025-08-21T18:48:34Z","timestamp":1755802114929,"version":"3.44.0"},"reference-count":23,"publisher":"Oxford University Press (OUP)","issue":"17","license":[{"start":{"date-parts":[[2025,6,24]],"date-time":"2025-06-24T00:00:00Z","timestamp":1750723200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,8,21]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Cofilin-2, encoded by CFL2, is an actin-binding protein essential for regulating actin filament dynamics in skeletal muscle. Biallelic variants in CFL2 are associated with an ultra-rare, early-onset myopathy typically presenting as nemaline myopathy. Only 10 patients have been described to date from five unrelated families. Here, we describe two new cases from two unrelated families. The first proband presented clinically with rigid spine syndrome and a biopsy keeping with nemaline myopathy. The second proband presented with a relatively mild congenital myopathy which became rapidly progressive in the fourth decade, the muscle biopsy showed cytoplasmic bodies, internal nuclei and ringbinden. Exome and genome sequencing revealed three novel biallelic missense variants in CFL2, a homozygous c.115\u00a0T\u2009&amp;gt;\u2009G; p.(Cys39Gly) in the proband of Family 1, and bi-allelic heterozygous c.256G\u2009&amp;gt;\u2009A:(p.Asp86Asn), and c.283A\u2009&amp;gt;\u2009G (p.Lys95Glu) variants in the proband of Family 2. We characterised the effects of these substitutions using an in vitro F-actin depolymerisation assay and showed all three were associated with significantly reduced filamentous actin depolymerisation rates compared to the wildtype. Taken together, our findings are highly suggestive of a CFL2-related disease in these patients. Since CFL2-related myopathies are ultrarare, the application of ACMG\/AMP guidelines and the diagnostic reportability of CFL2 variants identified in patients remains a challenge. The actin depolymerisation assay may be useful to elucidate the impact and pathogenicity of additional CFL2 variants and has the potential to guide variant classification in future.<\/jats:p>","DOI":"10.1093\/hmg\/ddaf102","type":"journal-article","created":{"date-parts":[[2025,6,28]],"date-time":"2025-06-28T23:17:36Z","timestamp":1751152656000},"page":"1471-1479","source":"Crossref","is-referenced-by-count":0,"title":["Novel missense variants in <i>CFL2<\/i> affect F-actin depolymerisation and expand the disease spectrum of <i>CFL2<\/i>-related myopathy"],"prefix":"10.1093","volume":"34","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4381-1909","authenticated-orcid":false,"given":"Lein N H","family":"Dofash","sequence":"first","affiliation":[{"name":"Harry Perkins Institute of Medical Research, Centre for Medical Research, University of Western Australia , Verdun Street, Nedlands, Perth WA 6009 ,","place":["Australia"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chiara","family":"Folland","sequence":"additional","affiliation":[{"name":"Harry Perkins Institute of Medical Research, Centre for Medical Research, University of Western Australia , Verdun Street, Nedlands, Perth WA 6009 ,","place":["Australia"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jason","family":"Dyke","sequence":"additional","affiliation":[{"name":"PathWest Neuropathology, Royal Perth Hospital, Victoria Square , Perth WA 6000 ,","place":["Australia"]},{"name":"School of Medicine and Pharmacology, University of Western Australia , Stirling Highway, Crawley, Perth WA 6009 ,","place":["Australia"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emna","family":"Farhat","sequence":"additional","affiliation":[{"name":"Faculty of Medicine of Tunis, University of Tunis El Manar , rue du Docteur Hassouna Ben Ayed, La Rabta, Tunis 1007 ,","place":["Tunisia"]},{"name":"The Muscle lab, Iris Medical Centre , Avenue H\u00e9di Nouira, Ennasr2, Ariana 2001 ,","place":["Tunisia"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Myriam","family":"Chaabouni","sequence":"additional","affiliation":[{"name":"Faculty of Medicine of Tunis, University of Tunis El Manar , rue du Docteur Hassouna Ben Ayed, La Rabta, Tunis 1007 ,","place":["Tunisia"]},{"name":"LGCgenetics, genetic laboratory, Les Jasmins Medical Centre, Centre Urbain Nord , Tunis 1082 ,","place":["Tunisia"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Najoua","family":"Miladi","sequence":"additional","affiliation":[{"name":"Faculty of Medicine of Tunis, University of Tunis El Manar , rue du Docteur Hassouna Ben Ayed, La Rabta, Tunis 1007 ,","place":["Tunisia"]},{"name":"Maghreb Medical Center , 14 Avenue du Roi Abdelaziz Al Saoud, El Manar 3, Tunis 2092 ,","place":["Tunisia"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Merrilee","family":"Needham","sequence":"additional","affiliation":[{"name":"Department of Neurology, Fiona Stanley Hospital , Robin Warren Dr, Murdoch, Perth WA 6150 ,","place":["Australia"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Phillipa J","family":"Lamont","sequence":"additional","affiliation":[{"name":"Neurogenetics Unit, Royal Perth Hospital , Victoria Square, Perth WA 6000 ,","place":["Australia"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8042-6180","authenticated-orcid":false,"given":"Catherine","family":"Ashton","sequence":"additional","affiliation":[{"name":"Neurogenetics Unit, Royal Perth Hospital , Victoria Square, Perth WA 6000 ,","place":["Australia"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gianina","family":"Ravenscroft","sequence":"additional","affiliation":[{"name":"Harry Perkins Institute of Medical Research, Centre for Medical Research, University of Western Australia , Verdun Street, Nedlands, Perth WA 6009 ,","place":["Australia"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2025,6,24]]},"reference":[{"key":"2025082106421563700_ref1","doi-asserted-by":"publisher","first-page":"743","DOI":"10.1126\/science.1094561","article-title":"Cofilin promotes actin polymerization and defines the direction of cell motility","volume":"304","author":"Ghosh","year":"2004","journal-title":"Science"},{"key":"2025082106421563700_ref2","doi-asserted-by":"publisher","first-page":"632","DOI":"10.1016\/j.nmd.2012.03.008","article-title":"Congenital myopathy caused by a novel missense mutation in the CFL2 gene","volume":"22","author":"Ockeloen","year":"2012","journal-title":"Neuromuscul Disord"},{"key":"2025082106421563700_ref3","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1091\/mbc.01-07-0331","article-title":"The three mouse actin-depolymerizing factor\/cofilins evolved to fulfill cell-type-specific requirements for actin dynamics","volume":"13","author":"Vartiainen","year":"2002","journal-title":"Mol Biol Cell"},{"key":"2025082106421563700_ref4","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1023\/A:1005682322132","article-title":"Expression of cofilin isoforms during development of mouse striated muscles","volume":"21","author":"Mohri","year":"2000","journal-title":"J Muscle Res Cell Motil"},{"key":"2025082106421563700_ref5","doi-asserted-by":"publisher","DOI":"10.3390\/cells12212523","article-title":"Role of actin-binding proteins in skeletal Myogenesis","volume":"12","author":"Nguyen","year":"2023","journal-title":"Cells"},{"key":"2025082106421563700_ref6","doi-asserted-by":"publisher","first-page":"2341","DOI":"10.1093\/hmg\/dds053","article-title":"Normal myofibrillar development followed by progressive sarcomeric disruption with actin accumulations in a mouse Cfl2 knockout demonstrates requirement of cofilin-2 for muscle maintenance","volume":"21","author":"Agrawal","year":"2012","journal-title":"Hum Mol Genet"},{"key":"2025082106421563700_ref7","doi-asserted-by":"publisher","first-page":"162","DOI":"10.1086\/510402","article-title":"Nemaline myopathy with minicores caused by mutation of the CFL2 gene encoding the skeletal muscle actin-binding protein, cofilin-2","volume":"80","author":"Agrawal","year":"2007","journal-title":"Am J Hum Genet"},{"key":"2025082106421563700_ref8","doi-asserted-by":"publisher","first-page":"1234","DOI":"10.1111\/cge.13240","article-title":"Expanding the histopathological spectrum of CFL2-related myopathies","volume":"93","author":"Fattori","year":"2018","journal-title":"Clin Genet"},{"key":"2025082106421563700_ref9","doi-asserted-by":"publisher","first-page":"1058","DOI":"10.1136\/jnnp-2014-307608","article-title":"Novel cofilin-2 (CFL2) four base pair deletion causing nemaline myopathy","volume":"85","author":"Ong","year":"2014","journal-title":"J Neurol Neurosurg Psychiatry"},{"key":"2025082106421563700_ref10","doi-asserted-by":"publisher","first-page":"252","DOI":"10.1016\/j.ejcb.2014.01.007","article-title":"Severe protein aggregate myopathy in a knockout mouse model points to an essential role of cofilin2 in sarcomeric actin exchange and muscle maintenance","volume":"93","author":"Gurniak","year":"2014","journal-title":"Eur J Cell Biol"},{"key":"2025082106421563700_ref11","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":"2025082106421563700_ref12","doi-asserted-by":"publisher","first-page":"6046","DOI":"10.1128\/MCB.00654-09","article-title":"Muscle LIM protein interacts with cofilin 2 and regulates F-actin dynamics in cardiac and skeletal muscle","volume":"29","author":"Papalouka","year":"2009","journal-title":"Mol Cell Biol"},{"key":"2025082106421563700_ref13","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1136\/jnnp-2014-309009","article-title":"Necklace cytoplasmic bodies in hereditary myopathy with early respiratory failure","volume":"86","author":"Uruha","year":"2015","journal-title":"J Neurol Neurosurg Psychiatry"},{"key":"2025082106421563700_ref14","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1016\/j.nmd.2017.02.012","article-title":"Cytoplasmic body pathology in severe ACTA1-related myopathy in the absence of typical nemaline rods","volume":"27","author":"Donkervoort","year":"2017","journal-title":"Neuromuscul Disord"},{"key":"2025082106421563700_ref15","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1186\/s40478-022-01400-0","article-title":"Severe ACTA1-related nemaline myopathy: intranuclear rods, cytoplasmic bodies, and enlarged perinuclear space as characteristic pathological features on muscle biopsies","volume":"10","author":"Labasse","year":"2022","journal-title":"Acta Neuropathol Commun"},{"key":"2025082106421563700_ref16","doi-asserted-by":"publisher","first-page":"911","DOI":"10.1006\/jmbi.2001.5280","article-title":"Determining the differences in actin binding by human ADF and cofilin","volume":"315","author":"Yeoh","year":"2002","journal-title":"J Mol Biol"},{"key":"2025082106421563700_ref17","doi-asserted-by":"publisher","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"},{"volume-title":"Muscle biopsy A Practical Approach","year":"2020","author":"Dubowitz","key":"2025082106421563700_ref18"},{"key":"2025082106421563700_ref19","doi-asserted-by":"crossref","first-page":"1127","DOI":"10.1093\/hmg\/ddac272","article-title":"A KLHL40 3' UTR splice-altering variant causes milder NEM8, an under-appreciated disease mechanism","volume":"32","author":"Dofash","year":"2022","journal-title":"Hum Mol Genet"},{"key":"2025082106421563700_ref20","doi-asserted-by":"publisher","first-page":"1276","DOI":"10.1038\/s41596-020-00462-5","article-title":"Detection of aberrant gene expression events in RNA sequencing data","volume":"16","author":"Y\u00e9pez","year":"2021","journal-title":"Nat Protoc"},{"key":"2025082106421563700_ref21","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1186\/s13073-022-01019-9","article-title":"Clinical implementation of RNA sequencing for Mendelian disease diagnostics","volume":"14","author":"Y\u00e9pez","year":"2022","journal-title":"Genome Med"},{"key":"2025082106421563700_ref22","doi-asserted-by":"publisher","first-page":"253","DOI":"10.3791\/253","article-title":"Transformation of plasmid DNA into E. Coli using the heat shock method","author":"Froger","year":"2007","journal-title":"J Vis Exp"},{"key":"2025082106421563700_ref23","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1385\/1-59259-584-7:531","article-title":"Protein identification and analysis tools in the ExPASy server","volume":"112","author":"Wilkins","year":"1999","journal-title":"Methods Mol Biol"}],"container-title":["Human Molecular Genetics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/hmg\/article-pdf\/34\/17\/1471\/63564665\/ddaf102.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/hmg\/article-pdf\/34\/17\/1471\/63564665\/ddaf102.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,8,21]],"date-time":"2025-08-21T10:42:26Z","timestamp":1755772946000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/hmg\/article\/34\/17\/1471\/8172346"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,24]]},"references-count":23,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2025,6,24]]},"published-print":{"date-parts":[[2025,8,21]]}},"URL":"https:\/\/doi.org\/10.1093\/hmg\/ddaf102","relation":{},"ISSN":["0964-6906","1460-2083"],"issn-type":[{"type":"print","value":"0964-6906"},{"type":"electronic","value":"1460-2083"}],"subject":[],"published-other":{"date-parts":[[2025,9,1]]},"published":{"date-parts":[[2025,6,24]]}}}