{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T05:03:52Z","timestamp":1789016632088,"version":"build-2803163510"},"reference-count":64,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2022,3,13]],"date-time":"2022-03-13T00:00:00Z","timestamp":1647129600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>A type 3 von Willebrand disease (VWD) index patient (IP) remains mutation-negative after completion of the conventional diagnostic analysis, including multiplex ligation-dependent probe amplification and sequencing of the promoter, exons, and flanking intronic regions of the VWF gene (VWF). In this study, we intended to elucidate causative mutation through next-generation sequencing (NGS) of the whole VWF (including complete intronic region), mRNA analysis, and study of the patient-derived endothelial colony-forming cells (ECFCs). The NGS revealed a variant in the intronic region of VWF (997 + 118 T &gt; G in intron 8), for the first time. The bioinformatics assessments (e.g., SpliceAl) predicted this variant creates a new donor splice site (ss), which could outcompete the consensus 5\u2032 donor ss at exon\/intron 8. This would lead to an aberrant mRNA that contains a premature stop codon, targeting it to nonsense-mediated mRNA decay. The subsequent quantitative real-time PCR confirmed the virtual absence of VWF mRNA in IP ECFCs. Additionally, the IP ECFCs demonstrated a considerable reduction in VWF secretion (~6% of healthy donors), and they were devoid of endothelial-specific secretory organelles, Weibel\u2013Palade bodies. Our findings underline the potential of NGS in conjunction with RNA analysis and patient-derived cell studies for genetic diagnosis of mutation-negative type 3 VWD patients.<\/jats:p>","DOI":"10.3390\/ijms23063095","type":"journal-article","created":{"date-parts":[[2022,3,13]],"date-time":"2022-03-13T22:29:43Z","timestamp":1647210583000},"page":"3095","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["A Homozygous Deep Intronic Variant Causes Von Willebrand Factor Deficiency and Lack of Endothelial-Specific Secretory Organelles, Weibel\u2013Palade Bodies"],"prefix":"10.3390","volume":"23","author":[{"given":"Hamideh","family":"Yadegari","sequence":"first","affiliation":[{"name":"Institute of Experimental Haematology and Transfusion Medicine, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad Ahmer","family":"Jamil","sequence":"additional","affiliation":[{"name":"Institute of Experimental Haematology and Transfusion Medicine, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natascha","family":"Marquardt","sequence":"additional","affiliation":[{"name":"Institute of Experimental Haematology and Transfusion Medicine, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1585-4100","authenticated-orcid":false,"given":"Johannes","family":"Oldenburg","sequence":"additional","affiliation":[{"name":"Institute of Experimental Haematology and Transfusion Medicine, University Hospital Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2019","DOI":"10.1182\/blood-2014-06-528406","article-title":"Von Willebrand factor biosynthesis, secretion, and clearance: Connecting the far ends","volume":"125","author":"Lenting","year":"2015","journal-title":"Blood"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1056\/NEJMc1616060","article-title":"Von Willebrand\u2019s Disease","volume":"376","author":"Leebeek","year":"2017","journal-title":"N. Engl. J. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3735","DOI":"10.1182\/blood-2013-06-498303","article-title":"Von Willebrand disease: Advances in pathogenetic understanding, diagnosis, and therapy","volume":"122","author":"Lillicrap","year":"2013","journal-title":"Blood"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"618","DOI":"10.1016\/j.thromres.2020.07.051","article-title":"How I treat von Willebrand disease","volume":"196","author":"Castaman","year":"2020","journal-title":"Thromb. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"280","DOI":"10.1182\/bloodadvances.2020003265","article-title":"ASH ISTH NHF WFH 2021 guidelines on the diagnosis of von Willebrand disease","volume":"5","author":"James","year":"2021","journal-title":"Blood Adv."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2103","DOI":"10.1111\/j.1538-7836.2006.02146.x","article-title":"Update on the pathophysiology and classification of von Willebrand disease: A report of the Subcommittee on von Willebrand Factor","volume":"4","author":"Sadler","year":"2006","journal-title":"J. Thromb. Haemost."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Pagliari, M.T., Rosendaal, F.R., Ahmadinejad, M., Badiee, Z., Baghaipour, M.-R., Baronciani, L., Ben\u00edtez Hidalgo, O., Bod\u00f3, I., Budde, U., and Castaman, G. (2022). Von Willebrand factor propeptide and pathophysiological mechanisms in European and Iranian patients with type 3 von Willebrand disease enrolled in the 3WINTERS-IPS study. J. Thromb. Haemost.","DOI":"10.1111\/jth.15658"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.thromres.2017.09.025","article-title":"Von Willebrand disease mutation spectrum and associated mutation mechanisms","volume":"159","author":"Eikenboom","year":"2017","journal-title":"Thromb. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"19514","DOI":"10.1016\/S0021-9258(19)47144-5","article-title":"Structure of the gene for human von Willebrand factor","volume":"264","author":"Mancuso","year":"1989","journal-title":"J. Biol. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1182\/blood-2012-01-405134","article-title":"Sequence and structure relationships within von Willebrand factor","volume":"120","author":"Zhou","year":"2012","journal-title":"Blood"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1002\/humu.24204","article-title":"Von Willebrand factor propeptide missense variants affect anterograde transport to Golgi resulting in ER retention","volume":"42","author":"Yadegari","year":"2021","journal-title":"Hum. Mutat."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"49982","DOI":"10.1074\/jbc.M408727200","article-title":"A covalent oxidoreductase intermediate in propeptide-dependent von Willebrand factor multimerization","volume":"279","author":"Purvis","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1785","DOI":"10.1182\/blood-2013-11-523639","article-title":"Highly reinforced structure of a C-terminal dimerization domain in von Willebrand factor","volume":"123","author":"Zhou","year":"2014","journal-title":"Blood"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3536","DOI":"10.1182\/blood.V83.12.3536.3536","article-title":"Von Willebrand factor proteolytic processing and multimerization precede the formation of Weibel-Palade bodies","volume":"83","author":"Vischer","year":"1994","journal-title":"Blood"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1183","DOI":"10.1182\/blood-2015-04-641902","article-title":"Von Willebrand factor is dimerized by protein disulfide isomerase","volume":"127","author":"Lippok","year":"2016","journal-title":"Blood"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5033","DOI":"10.1182\/blood-2010-09-267492","article-title":"Functional architecture of Weibel-Palade bodies","volume":"117","author":"Valentijn","year":"2011","journal-title":"Blood"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"718","DOI":"10.1002\/rth2.12242","article-title":"Alternative trafficking of Weibel-Palade body proteins in CRISPR\/Cas9-engineered von Willebrand factor-deficient blood outgrowth endothelial cells","volume":"3","author":"Schillemans","year":"2019","journal-title":"Res. Pract. Thromb. Haemost."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1873","DOI":"10.1111\/jth.14218","article-title":"Tuning the endothelial response: Differential release of exocytic cargos from Weibel-Palade bodies","volume":"16","author":"Nightingale","year":"2018","journal-title":"J. Thromb. Haemost."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"3611","DOI":"10.1242\/jcs.208033","article-title":"Weibel-Palade bodies at a glance","volume":"130","author":"McCormack","year":"2017","journal-title":"J. Cell Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"662","DOI":"10.1160\/TH12-02-0089","article-title":"Mutation distribution in the von Willebrand factor gene related to the different von Willebrand disease (VWD) types in a cohort of VWD patients","volume":"108","author":"Yadegari","year":"2012","journal-title":"Thromb. Haemost."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"652","DOI":"10.1160\/TH12-10-0737","article-title":"Phenotypic and molecular characterisation of type 3 von Willebrand disease in a cohort of Indian patients","volume":"109","author":"Ahmad","year":"2013","journal-title":"Thromb. Haemost."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1016\/S1079-9796(03)00033-0","article-title":"Molecular defects in type 3 von Willebrand disease: Updated results from 40 multiethnic patients","volume":"30","author":"Baronciani","year":"2003","journal-title":"Blood Cells Mol. Dis."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"512","DOI":"10.1111\/jth.12130","article-title":"The genetics of Canadian type 3 von Willebrand disease: Further evidence for co-dominant inheritance of mutant alleles","volume":"11","author":"Bowman","year":"2013","journal-title":"J. Thromb. Haemost."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/j.bcmd.2008.03.004","article-title":"Genetic defects in von Willebrand disease type 3 in Indian and Greek patients","volume":"41","author":"Gupta","year":"2008","journal-title":"Blood Cells Mol. Dis."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1048","DOI":"10.1111\/j.1365-2516.2009.02059.x","article-title":"The mutation spectrum associated with type 3 von Willebrand disease in a cohort of patients from the North West of England","volume":"15","author":"Sutherland","year":"2009","journal-title":"Haemophilia"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1035","DOI":"10.1111\/hae.13841","article-title":"Characterization of the mutation spectrum in a Pakistani cohort of type 3 von Willebrand disease","volume":"25","author":"Ahmed","year":"2019","journal-title":"Haemophilia"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1083","DOI":"10.1111\/j.1538-7836.2011.04260.x","article-title":"Large deletions identified in patients with von Willebrand disease using multiple ligation-dependent probe amplification","volume":"9","author":"Yadegari","year":"2011","journal-title":"J. Thromb. Haemost."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1055\/s-0039-3400260","article-title":"The Current Understanding of Molecular Pathogenesis of Quantitative von Willebrand Disease, Types 1 and 3","volume":"40","author":"Yadegari","year":"2020","journal-title":"Hamostaseologie"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e297","DOI":"10.1097\/HS9.0000000000000297","article-title":"Von Willebrand Disease: From In Vivo to In Vitro Disease Models","volume":"3","author":"Eikenboom","year":"2019","journal-title":"Hemasphere"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1002\/humu.24312","article-title":"Unraveling the molecular basis underlying nine putative splice site variants of von Willebrand factor","volume":"43","author":"Liang","year":"2021","journal-title":"Hum. Mutat."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"584","DOI":"10.1182\/blood-2015-10-678052","article-title":"Characterization of aberrant splicing of von Willebrand factor in von Willebrand disease: An underrecognized mechanism","volume":"128","author":"Hawke","year":"2016","journal-title":"Blood"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1160\/TH06-07-0417","article-title":"Combined partial exon skipping and cryptic splice site activation as a new molecular mechanism for recessive type 1 von Willebrand disease","volume":"96","author":"Gallinaro","year":"2006","journal-title":"Thromb. Haemost."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2144","DOI":"10.1182\/blood-2016-02-699686","article-title":"Intron retention resulting from a silent mutation in the VWF gene that structurally influences the 5\u2019 splice site","volume":"128","author":"Yadegari","year":"2016","journal-title":"Blood"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Conboy, J.G. (2021). A Deep Exon Cryptic Splice Site Promotes Aberrant Intron Retention in a Von Willebrand Disease Patient. Int. J. Mol. Sci., 22.","DOI":"10.1101\/2021.11.02.466821"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.1007\/s00439-017-1809-4","article-title":"Deep intronic mutations and human disease","volume":"136","year":"2017","journal-title":"Hum. Genet."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1411","DOI":"10.1038\/s41598-017-01577-8","article-title":"A deep intronic CLRN1 (USH3A) founder mutation generates an aberrant exon and underlies severe Usher syndrome on the Arabian Peninsula","volume":"7","author":"Khan","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"660113","DOI":"10.3389\/fneur.2021.660113","article-title":"A Homozygous Deep Intronic Mutation Alters the Splicing of Nebulin Gene in a Patient With Nemaline Myopathy","volume":"12","author":"Laflamme","year":"2021","journal-title":"Front. Neurol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e1090","DOI":"10.1002\/mgg3.1090","article-title":"A novel mutation deep within intron 7 of the GBA gene causes Gaucher disease","volume":"8","author":"Malekkou","year":"2020","journal-title":"Mol. Genet. Genom. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1007\/s10875-020-00753-2","article-title":"Deep Intronic Mutation in SERPING1 Caused Hereditary Angioedema Through Pseudoexon Activation","volume":"40","author":"Hakl","year":"2020","journal-title":"J. Clin. Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"276","DOI":"10.3389\/fgene.2021.647400","article-title":"Identification of Deep-Intronic Splice Mutations in a Large Cohort of Patients With Inherited Retinal Diseases","volume":"12","author":"Qian","year":"2021","journal-title":"Front. Genet."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"18097","DOI":"10.1038\/s41598-021-97414-0","article-title":"mRNA analysis identifies deep intronic variants causing Alport syndrome and overcomes the problem of negative results of exome sequencing","volume":"11","author":"Wang","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1679","DOI":"10.1111\/jth.12339","article-title":"Deep intronic \u2018mutations\u2019 cause hemophilia A: Application of next generation sequencing in patients without detectable mutation in F8 cDNA","volume":"11","author":"Pezeshkpoor","year":"2013","journal-title":"J. Thromb. Haemost."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1854","DOI":"10.1111\/jth.13112","article-title":"Angiogenic characteristics of blood outgrowth endothelial cells from patients with von Willebrand disease","volume":"13","author":"Groeneveld","year":"2015","journal-title":"J. Thromb. Haemost."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1071","DOI":"10.1182\/blood-2010-01-264507","article-title":"Endothelial von Willebrand factor regulates angiogenesis","volume":"117","author":"Starke","year":"2011","journal-title":"Blood"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"4072","DOI":"10.1073\/pnas.061307098","article-title":"Defect in regulated secretion of P-selectin affects leukocyte recruitment in von Willebrand factor-deficient mice","volume":"98","author":"Denis","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1182\/blood-2004-02-0464","article-title":"Re-establishment of VWF-dependent Weibel-Palade bodies in VWD endothelial cells","volume":"105","author":"Haberichter","year":"2005","journal-title":"Blood"},{"key":"ref_47","first-page":"1471","article-title":"Heterogeneous distribution of Weibel-Palade bodies and von Willebrand factor along the porcine vascular tree","volume":"139","author":"Drouet","year":"1991","journal-title":"Am. J. Pathol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1038","DOI":"10.1182\/bloodadvances.2021005895","article-title":"Multifaceted Pathomolecular Mechanism of a VWF Large Deletion Involved in the Pathogenesis of Severe VWD","volume":"6","author":"Yadegari","year":"2022","journal-title":"Blood Adv."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"a006550","DOI":"10.1101\/cshperspect.a006650","article-title":"The complex role of angiopoietin-2 in the angiopoietin-tie signaling pathway","volume":"2","author":"Thurston","year":"2012","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"3495","DOI":"10.1172\/JCI84923","article-title":"Tie1 controls angiopoietin function in vascular remodeling and inflammation","volume":"126","author":"Korhonen","year":"2016","journal-title":"J. Clin. Investig."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"4150","DOI":"10.1182\/blood-2003-10-3685","article-title":"The Tie-2 ligand angiopoietin-2 is stored in and rapidly released upon stimulation from endothelial cell Weibel-Palade bodies","volume":"103","author":"Fiedler","year":"2004","journal-title":"Blood"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1161\/ATVBAHA.108.172676","article-title":"Angiopoietin-2 exocytosis is stimulated by sphingosine-1-phosphate in human blood and lymphatic endothelial cells","volume":"29","author":"Jang","year":"2009","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"2063","DOI":"10.1111\/j.1538-7836.2010.03975.x","article-title":"ISTH\/SSC bleeding assessment tool: A standardized questionnaire and a proposal for a new bleeding score for inherited bleeding disorders","volume":"8","author":"Rodeghiero","year":"2010","journal-title":"J. Thromb. Haemost."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1186\/s13059-016-0974-4","article-title":"The Ensembl Variant Effect Predictor","volume":"17","author":"McLaren","year":"2016","journal-title":"Genome Biol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1016\/j.cell.2018.12.015","article-title":"Predicting Splicing from Primary Sequence with Deep Learning","volume":"176","author":"Jaganathan","year":"2019","journal-title":"Cell"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1089\/cmb.1997.4.311","article-title":"Improved splice site detection in Genie","volume":"4","author":"Reese","year":"1997","journal-title":"J. Comput. Biol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/j.gene.2005.07.015","article-title":"Characterization and prediction of alternative splice sites","volume":"366","author":"Wang","year":"2006","journal-title":"Gene"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1709","DOI":"10.1038\/nprot.2012.093","article-title":"Establishment of outgrowth endothelial cells from peripheral blood","volume":"7","author":"Hofman","year":"2012","journal-title":"Nat. Protoc."},{"key":"ref_59","doi-asserted-by":"crossref","unstructured":"Ormiston, M.L., Toshner, M.R., Kiskin, F.N., Huang, C.J.Z., Groves, E., Morrell, N.W., and Rana, A.A. (2015). Generation and Culture of Blood Outgrowth Endothelial Cells from Human Peripheral Blood. J. Vis. Exp., e53384.","DOI":"10.3791\/53384"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"25749","DOI":"10.1038\/srep25749","article-title":"miARma-Seq: A comprehensive tool for miRNA, mRNA and circRNA analysis","volume":"6","author":"Rojas","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"R36","DOI":"10.1186\/gb-2013-14-4-r36","article-title":"TopHat2: Accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions","volume":"14","author":"Kim","year":"2013","journal-title":"Genome Biol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"10","DOI":"10.14806\/ej.17.1.200","article-title":"Cutadapt removes adapter sequences from high-throughput sequencing reads","volume":"17","author":"Martin","year":"2011","journal-title":"EMBnet J."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1038\/nmeth.1923","article-title":"Fast gapped-read alignment with Bowtie 2","volume":"9","author":"Langmead","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"1315","DOI":"10.3324\/haematol.2013.084111","article-title":"Insights into pathological mechanisms of missense mutations in C-terminal domains of von Willebrand factor causing qualitative or quantitative von Willebrand disease","volume":"98","author":"Yadegari","year":"2013","journal-title":"Haematologica"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/6\/3095\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:35:51Z","timestamp":1760135751000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/6\/3095"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,13]]},"references-count":64,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["ijms23063095"],"URL":"https:\/\/doi.org\/10.3390\/ijms23063095","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,13]]}}}