{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T12:26:19Z","timestamp":1770985579434,"version":"3.50.1"},"reference-count":52,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2021,4,13]],"date-time":"2021-04-13T00:00:00Z","timestamp":1618272000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP20K09867"],"award-info":[{"award-number":["JP20K09867"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP19H04754"],"award-info":[{"award-number":["JP19H04754"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Congenital hearing loss affects 1 in every 1000 births, with genetic mutations contributing to more than 50% of all cases. X-linked nonsyndromic hereditary hearing loss is associated with six loci (DFNX1-6) and five genes. Recently, the missense mutation (c.1771G&gt;A, p.Gly591Ser) in<jats:italic>COL4A6<\/jats:italic>, encoding the basement membrane (BM) collagen \u03b16(IV) chain, was shown to be associated with X-linked congenital nonsyndromic hearing loss with cochlear malformation. However, the mechanism by which the<jats:italic>COL4A6<\/jats:italic>mutation impacts hereditary hearing loss has not yet been elucidated. Herein, we investigated<jats:italic>Col4a6<\/jats:italic>knockout (KO) effects on hearing function and cochlear formation in mice. Immunohistochemistry showed that the collagen \u03b16(IV) chain was distributed throughout the mouse cochlea within subepithelial BMs underlying the interdental cells, inner sulcus cells, basilar membrane, outer sulcus cells, root cells, Reissner\u2019s membrane, and perivascular BMs in the spiral limbus, spiral ligament, and stria vascularis. However, the click-evoked auditory brainstem response analysis did not show significant changes in the hearing threshold of<jats:italic>Col4a6<\/jats:italic>KO mice compared with wild-type (WT) mice with the same genetic background. In addition, the cochlear structures of<jats:italic>Col4a6<\/jats:italic>KO mice did not exhibit morphological alterations, according to the results of high-resolution micro-computed tomography and histology. Hence, loss of<jats:italic>Col4a6<\/jats:italic>gene expression in mice showed normal click ABR thresholds and normal cochlear formation, which differs from humans with the<jats:italic>COL4A6<\/jats:italic>missense mutation c.1771G&gt;A, p.Gly591Ser. Therefore, the deleterious effects in the auditory system caused by the missense mutation in<jats:italic>COL4A6<\/jats:italic>are likely due to the dominant-negative effects of the \u03b16(IV) chain and\/or \u03b15\u03b16\u03b15(IV) heterotrimer with an aberrant structure that would not occur in cases with loss of gene expression.<\/jats:p>","DOI":"10.1371\/journal.pone.0249909","type":"journal-article","created":{"date-parts":[[2021,4,13]],"date-time":"2021-04-13T17:57:19Z","timestamp":1618336639000},"page":"e0249909","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":5,"title":["Lack of collagen \u03b16(IV) chain in mice does not cause severe-to-profound hearing loss or cochlear malformation, a distinct phenotype from nonsyndromic hearing loss with COL4A6 missense mutation"],"prefix":"10.1371","volume":"16","author":[{"given":"Shaoying","family":"Tang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3380-8298","authenticated-orcid":true,"given":"Tomoko","family":"Yonezawa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yukihide","family":"Maeda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mitsuaki","family":"Ono","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takahiro","family":"Maeba","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toru","family":"Miyoshi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ryusuke","family":"Momota","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yasuko","family":"Tomono","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toshitaka","family":"Oohashi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2021,4,13]]},"reference":[{"key":"pone.0249909.ref001","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/bs.ctdb.2015.07.007","article-title":"Development and Integration of the Ear","volume":"115","author":"JC Fuchs","year":"2015","journal-title":"Curr Top Dev Biol"},{"issue":"2","key":"pone.0249909.ref002","doi-asserted-by":"crossref","first-page":"324","DOI":"10.1111\/joa.12308","article-title":"Form and function of the mammalian inner ear","volume":"228","author":"EG Ekdale","year":"2016","journal-title":"J Anat"},{"key":"pone.0249909.ref003","doi-asserted-by":"crossref","first-page":"580639","DOI":"10.3389\/fneur.2020.580639","article-title":"On the Role of Fibrocytes and the Extracellular Matrix in the Physiology and Pathophysiology of the Spiral Ligament.","volume":"11","author":"N Peeleman","year":"2020","journal-title":"Front Neurol."},{"issue":"4","key":"pone.0249909.ref004","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1016\/j.chom.2019.02.001","article-title":"Cleaved Cochlin Sequesters Pseudomonas aeruginosa and Activates Innate Immunity in the Inner Ear","volume":"25","author":"J Jung","year":"2019","journal-title":"Cell Host Microbe"},{"key":"pone.0249909.ref005","first-page":"57","article-title":"The nature and biology of basement membranes","author":"A Pozzi","year":"2017","journal-title":"Matrix Biol"},{"issue":"5","key":"pone.0249909.ref006","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1093\/oxfordjournals.jbchem.a022003","article-title":"Organization and expression of basement membrane collagen IV genes and their roles in human disorders","volume":"123","author":"Y Sado","year":"1998","journal-title":"J Biochem"},{"issue":"5","key":"pone.0249909.ref007","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1002\/jemt.20564","article-title":"Mammalian collagen IV","volume":"71","author":"J Khoshnoodi","year":"2008","journal-title":"Microsc Res Tech"},{"issue":"11","key":"pone.0249909.ref008","doi-asserted-by":"crossref","first-page":"2151","DOI":"10.1002\/pro.3283","article-title":"Building collagen IV smart scaffolds on the outside of cells","volume":"26","author":"KL Brown","year":"2017","journal-title":"Protein Sci"},{"issue":"5","key":"pone.0249909.ref009","doi-asserted-by":"crossref","first-page":"1219","DOI":"10.1083\/jcb.130.5.1219","article-title":"Differential expression of two basement membrane collagen genes, COL4A6 and COL4A5, demonstrated by immunofluorescence staining using peptide-specific monoclonal antibodies","volume":"130","author":"Y Ninomiya","year":"1995","journal-title":"J Cell Biol"},{"issue":"4","key":"pone.0249909.ref010","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1007\/s004180050296","article-title":"Differential expression of type IV collagen isoforms, alpha5(IV) and alpha6(IV) chains, in basement membranes surrounding smooth muscle cells.","volume":"110","author":"T Seki","year":"1998","journal-title":"Histochem Cell Biol"},{"issue":"3","key":"pone.0249909.ref011","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1093\/oxfordjournals.jbchem.a022770","article-title":"Differential expression of mouse alpha5(IV) and alpha6(IV) collagen genes in epithelial basement membranes.","volume":"128","author":"K Saito","year":"2000","journal-title":"Journal of biochemistry"},{"issue":"30","key":"pone.0249909.ref012","doi-asserted-by":"crossref","first-page":"28532","DOI":"10.1074\/jbc.M103690200","article-title":"The NC1 domain of collagen IV encodes a novel network composed of the alpha 1, alpha 2, alpha 5, and alpha 6 chains in smooth muscle basement membranes","volume":"276","author":"DB Borza","year":"2001","journal-title":"J Biol Chem"},{"issue":"2","key":"pone.0249909.ref013","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1679\/aohc.65.133","article-title":"Basement membrane type IV collagen molecules in the choroid plexus, pia mater and capillaries in the mouse brain","volume":"65","author":"N Urabe","year":"2002","journal-title":"Arch Histol Cytol"},{"issue":"4","key":"pone.0249909.ref014","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1679\/aohc.70.255","article-title":"The distributions of type IV collagen alpha chains in basement membranes of human epidermis and skin appendages.","volume":"70","author":"H Hasegawa","year":"2007","journal-title":"Arch Histol Cytol"},{"issue":"2","key":"pone.0249909.ref015","doi-asserted-by":"crossref","first-page":"147","DOI":"10.3109\/03008207.2010.492062","article-title":"Distribution of alpha(IV) collagen chains in the ocular anterior segments of adult mice.","volume":"52","author":"K Saito","year":"2011","journal-title":"Connective tissue research"},{"issue":"10","key":"pone.0249909.ref016","doi-asserted-by":"crossref","first-page":"7520","DOI":"10.1016\/S0021-9258(17)37317-9","article-title":"Identification of a new collagen IV chain, alpha 6(IV), by cDNA isolation and assignment of the gene to chromosome Xq22, which is the same locus for COL4A5.","volume":"269","author":"T Oohashi","year":"1994","journal-title":"J Biol Chem"},{"issue":"18","key":"pone.0249909.ref017","doi-asserted-by":"crossref","first-page":"13193","DOI":"10.1016\/S0021-9258(17)36818-7","article-title":"Complete primary structure of the sixth chain of human basement membrane collagen, alpha 6(IV). Isolation of the cDNAs for alpha 6(IV) and comparison with five other type IV collagen chains.","volume":"269","author":"J Zhou","year":"1994","journal-title":"J Biol Chem"},{"issue":"1","key":"pone.0249909.ref018","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/j.cell.2007.02.035","article-title":"Distinct target-derived signals organize formation, maturation, and maintenance of motor nerve terminals","volume":"129","author":"MA Fox","year":"2007","journal-title":"Cell"},{"key":"pone.0249909.ref019","doi-asserted-by":"crossref","first-page":"29450","DOI":"10.1038\/srep29450","article-title":"COL4A6 is dispensable for autosomal recessive Alport syndrome.","volume":"6","author":"T Murata","year":"2016","journal-title":"Sci Rep."},{"issue":"1","key":"pone.0249909.ref020","doi-asserted-by":"crossref","first-page":"2612","DOI":"10.1038\/s41598-018-21000-0","article-title":"Type IV collagen alpha6 chain is a regulator of keratin 10 in keratinization of oral mucosal epithelium.","volume":"8","author":"T Komori","year":"2018","journal-title":"Sci Rep."},{"issue":"10","key":"pone.0249909.ref021","doi-asserted-by":"crossref","first-page":"e1005587","DOI":"10.1371\/journal.pgen.1005587","article-title":"Type IV Collagen Controls the Axogenesis of Cerebellar Granule Cells by Regulating Basement Membrane Integrity in Zebrafish.","volume":"11","author":"M Takeuchi","year":"2015","journal-title":"PLoS Genet."},{"issue":"2","key":"pone.0249909.ref022","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1038\/ejhg.2013.108","article-title":"Novel form of X-linked nonsyndromic hearing loss with cochlear malformation caused by a mutation in the type IV collagen gene COL4A6","volume":"22","author":"S Rost","year":"2014","journal-title":"Eur J Hum Genet"},{"issue":"1","key":"pone.0249909.ref023","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1111\/j.1523-1755.2004.00719.x","article-title":"High nephritogenicity of monoclonal antibodies belonging to IgG2a and IgG2b subclasses in rat anti-GBM nephritis","volume":"66","author":"T Kohda","year":"2004","journal-title":"Kidney Int"},{"issue":"11","key":"pone.0249909.ref024","doi-asserted-by":"crossref","first-page":"3137","DOI":"10.1002\/j.1460-2075.1991.tb04875.x","article-title":"Recombinant nidogen consists of three globular domains and mediates binding of laminin to collagen type IV","volume":"10","author":"JW Fox","year":"1991","journal-title":"Embo j"},{"issue":"3","key":"pone.0249909.ref025","doi-asserted-by":"crossref","first-page":"e58775","DOI":"10.1371\/journal.pone.0058775","article-title":"Time courses of changes in phospho- and total- MAP kinases in the cochlea after intense noise exposure.","volume":"8","author":"Y Maeda","year":"2013","journal-title":"PLoS One"},{"issue":"7","key":"pone.0249909.ref026","doi-asserted-by":"crossref","first-page":"e593","DOI":"10.1097\/MAO.0000000000001874","article-title":"Targeted PCR Array Analysis of Genes in Innate Immunity and Glucocorticoid Signaling Pathways in Mice Cochleae Following Acoustic Trauma","volume":"39","author":"Y Maeda","year":"2018","journal-title":"Otol Neurotol"},{"key":"pone.0249909.ref027","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/j.bone.2014.09.011","article-title":"The BMP2 antagonist inhibitor L51P enhances the osteogenic potential of BMP2 by simultaneous and delayed synergism","volume":"69","author":"HM Khattab","year":"2014","journal-title":"Bone"},{"issue":"1\u20132","key":"pone.0249909.ref028","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/S0378-5955(98)00069-0","article-title":"Ultrastructural, physiological, and molecular defects in the inner ear of a gene-knockout mouse model for autosomal Alport syndrome","volume":"121","author":"D Cosgrove","year":"1998","journal-title":"Hear Res"},{"issue":"1\u20132","key":"pone.0249909.ref029","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/S0378-5955(01)00278-7","article-title":"Proteoglycan arrays in the cochlear basement membrane","volume":"157","author":"V Tsuprun","year":"2001","journal-title":"Hear Res"},{"issue":"1\u20132","key":"pone.0249909.ref030","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/S0378-5955(96)80007-4","article-title":"Immunohistochemical localization of basement membrane collagens and associated proteins in the murine cochlea","volume":"97","author":"D Cosgrove","year":"1996","journal-title":"Hear Res"},{"issue":"4","key":"pone.0249909.ref031","doi-asserted-by":"crossref","first-page":"239","DOI":"10.2220\/biomedres.38.239","article-title":"Three-dimensional reconstruction of root cells and interdental cells in the rat inner ear by serial section scanning electron microscopy","volume":"38","author":"R Shodo","year":"2017","journal-title":"Biomed Res"},{"issue":"4","key":"pone.0249909.ref032","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1007\/BF01464322","article-title":"Establishment by the rat lymph node method of epitope-defined monoclonal antibodies recognizing the six different alpha chains of human type IV collagen","volume":"104","author":"Y Sado","year":"1995","journal-title":"Histochem Cell Biol"},{"issue":"4","key":"pone.0249909.ref033","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1007\/s00424-009-0754-z","article-title":"How is the highly positive endocochlear potential formed? The specific architecture of the stria vascularis and the roles of the ion-transport apparatus","volume":"459","author":"H Hibino","year":"2010","journal-title":"Pflugers Arch"},{"issue":"1\u20132","key":"pone.0249909.ref034","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/S0378-5955(01)00252-0","article-title":"Three-dimensional and ultrastructural relationships between intermediate cells and capillaries in the gerbil stria vascularis","volume":"155","author":"S Takeuchi","year":"2001","journal-title":"Hear Res"},{"issue":"3","key":"pone.0249909.ref035","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1002\/glia.10279","article-title":"Limitrin, a novel immunoglobulin superfamily protein localized to glia limitans formed by astrocyte endfeet","volume":"44","author":"T Yonezawa","year":"2003","journal-title":"Glia"},{"issue":"7","key":"pone.0249909.ref036","doi-asserted-by":"crossref","first-page":"755","DOI":"10.1002\/glia.20959","article-title":"Type IV collagen induces expression of thrombospondin-1 that is mediated by integrin alpha1beta1 in astrocytes","volume":"58","author":"T Yonezawa","year":"2010","journal-title":"Glia"},{"issue":"1","key":"pone.0249909.ref037","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1007\/s00405-014-3478-4","article-title":"Age-related morphological changes in the basement membrane in the stria vascularis of C57BL\/6 mice.","volume":"273","author":"M Suzuki","year":"2016","journal-title":"Eur Arch Otorhinolaryngol"},{"key":"pone.0249909.ref038","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.heares.2016.08.003","article-title":"Endothelin-1 mediated induction of extracellular matrix genes in strial marginal cells underlies strial pathology in Alport mice","volume":"341","author":"DT Meehan","year":"2016","journal-title":"Hear Res"},{"issue":"10","key":"pone.0249909.ref039","doi-asserted-by":"crossref","first-page":"1903","DOI":"10.3174\/ajnr.A4822","article-title":"Blood-Labyrinth Barrier Permeability in Meni\u00e8re Disease and Idiopathic Sudden Sensorineural Hearing Loss: Findings on Delayed Postcontrast 3D-FLAIR MRI.","volume":"37","author":"MN Pakdaman","year":"2016","journal-title":"AJNR Am J Neuroradiol."},{"key":"pone.0249909.ref040","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.heares.2016.01.010","article-title":"Pathophysiology of the cochlear intrastrial fluid-blood barrier (review).","volume":"338","author":"X. Shi","year":"2016","journal-title":"Hear Res"},{"issue":"1","key":"pone.0249909.ref041","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1038\/s41598-017-00330-5","article-title":"The blood labyrinthine barrier in the human normal and Meniere\u2019s disease macula utricle","volume":"7","author":"G Ishiyama","year":"2017","journal-title":"Sci Rep"},{"issue":"4","key":"pone.0249909.ref042","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1038\/70516","article-title":"Mutations in COL11A2 cause non-syndromic hearing loss (DFNA13).","volume":"23","author":"WT McGuirt","year":"1999","journal-title":"Nat Genet"},{"issue":"4","key":"pone.0249909.ref043","doi-asserted-by":"crossref","first-page":"948","DOI":"10.1038\/s41436-018-0285-0","article-title":"Splice-altering variant in COL11A1 as a cause of nonsyndromic hearing loss DFNA37","volume":"21","author":"KT Booth","year":"2019","journal-title":"Genet Med"},{"issue":"3","key":"pone.0249909.ref044","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1002\/humu.20429","article-title":"Consortium for osteogenesis imperfecta mutations in the helical domain of type I collagen: regions rich in lethal mutations align with collagen binding sites for integrins and proteoglycans","volume":"28","author":"JC Marini","year":"2007","journal-title":"Hum Mutat"},{"issue":"9","key":"pone.0249909.ref045","doi-asserted-by":"crossref","first-page":"540","DOI":"10.1038\/nrendo.2011.81","article-title":"New perspectives on osteogenesis imperfecta.","volume":"7","author":"A Forlino","year":"2011","journal-title":"Nat Rev Endocrinol"},{"issue":"7","key":"pone.0249909.ref046","doi-asserted-by":"crossref","first-page":"1218","DOI":"10.1093\/ndt\/17.7.1218","article-title":"Meta-analysis of genotype-phenotype correlation in X-linked Alport syndrome: impact on clinical counselling.","volume":"17","author":"O Gross","year":"2002","journal-title":"Nephrol Dial Transplant"},{"issue":"5","key":"pone.0249909.ref047","doi-asserted-by":"crossref","first-page":"1208","DOI":"10.1038\/ki.2013.479","article-title":"Milder clinical aspects of X-linked Alport syndrome in men positive for the collagen IV alpha5 chain","volume":"85","author":"Y Hashimura","year":"2014","journal-title":"Kidney Int"},{"key":"pone.0249909.ref048","article-title":"Alport syndrome and Pierson syndrome: Diseases of the glomerular basement membrane","author":"SD Funk","year":"2018","journal-title":"Matrix Biol"},{"issue":"5","key":"pone.0249909.ref049","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1007\/s00467-019-04241-7","article-title":"The importance of clinician, patient and researcher collaborations in Alport syndrome","volume":"35","author":"MN Rheault","year":"2020","journal-title":"Pediatr Nephrol"},{"issue":"1","key":"pone.0249909.ref050","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1038\/ki.1996.316","article-title":"Relationship between COL4A5 gene mutation and distribution of type IV collagen in male X-linked Alport syndrome. Japanese Alport Network","volume":"50","author":"I Naito","year":"1996","journal-title":"Kidney Int"},{"issue":"2","key":"pone.0249909.ref051","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1681\/ASN.2012121217","article-title":"Evidence for activation of the unfolded protein response in collagen IV nephropathies","volume":"25","author":"M Pieri","year":"2014","journal-title":"J Am Soc Nephrol"},{"issue":"5","key":"pone.0249909.ref052","doi-asserted-by":"crossref","first-page":"634","DOI":"10.1016\/j.chembiol.2018.02.003","article-title":"A Split-Luciferase-Based Trimer Formation Assay as a High-throughput Screening Platform for Therapeutics in Alport Syndrome.","volume":"25","author":"K Omachi","year":"2018","journal-title":"Cell Chem Biol"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0249909","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,8,28]],"date-time":"2024-08-28T07:08:23Z","timestamp":1724828903000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0249909"}},"subtitle":[],"editor":[{"given":"Olivia","family":"Bermingham-McDonogh","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2021,4,13]]},"references-count":52,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2021,4,13]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0249909","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,13]]}}}