{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,2]],"date-time":"2025-11-02T04:12:04Z","timestamp":1762056724640,"version":"build-2065373602"},"reference-count":41,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2022,6,23]],"date-time":"2022-06-23T00:00:00Z","timestamp":1655942400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Deanship of Scientific Research, King Saud University for funding through Vice Deanship of Scientific Research Chairs","award":["KMU\/ORIC\/ar\/007"],"award-info":[{"award-number":["KMU\/ORIC\/ar\/007"]}]},{"name":"Khyber Medical University","award":["KMU\/ORIC\/ar\/007"],"award-info":[{"award-number":["KMU\/ORIC\/ar\/007"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background: Dystrophic Epidermolysis bullosa (DEB) is a rare, severe subtype of epidermolysis bullosa (EB), characterized by blisters and miliary rashes of the skin. Dystrophic EB (DEB) includes variants inherited both in an autosomal-dominant or autosomal-recessive manner. Recessive dystrophic EB (RDEB) is divided into many subtypes and prevails as a result of biallelic genetic mutations in COL7A1 gene encoding type VII collagen, a major stabilizing molecule of the dermo-epidermal junction. The blister formation is mainly due to the variable structural and functional impairment of anchoring fibrils in VII collagen (COLVII), responsible for the adhesion of the epidermis to the dermis. Method: Three Pakistani families (A, B and C) affected with congenital dystrophic epidermolysis bullosa were recruited in the present study. The whole-exome sequencing (WES) approach was utilized for the detection of the pathogenic sequence variants in probands. The segregation of these variants in other participants was confirmed by Sanger sequencing. Results: This study identified a novel missense variant c.7034G&gt;A, p. Gly2345Asp in exon 91, a novel Frameshift mutation c.385del (p. His129MetfsTer18) in a homozygous form in exon no 3, and a previously known nonsense variation (c.1573 C&gt;T; p. Arg525Ter) in exon 12 of COL7A1 gene in families A, B, and C, respectively, as causative mutations responsible for dystrophic epidermolysis bullosa in these families. Conclusion: Our study validates the involvement of the COL7A1 gene in the etiology of dystrophic epidermolysis bullosa. It further expands the COL7A1 gene mutation database and provides an additional scientific basis for diagnosis, genetic counseling, and prognosis purposes for EB patients.<\/jats:p>","DOI":"10.3390\/diagnostics12071525","type":"journal-article","created":{"date-parts":[[2022,6,23]],"date-time":"2022-06-23T01:59:47Z","timestamp":1655949587000},"page":"1525","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Detection of Novel Biallelic Causative Variants in COL7A1 Gene by Whole-Exome Sequencing, Resulting in Congenital Recessive Dystrophic Epidermolysis Bullosa in Three Unrelated Families"],"prefix":"10.3390","volume":"12","author":[{"given":"Fozia","family":"Fozia","sequence":"first","affiliation":[{"name":"Institute of Basic Medical Sciences (IBMS), Khyber Medical University (KMU), Peshawar 25100, Khyber Pakhtunkhwa, Pakistan"},{"name":"Department of Biotechnology and Genetic Engineering, Kohat University of Science & Technology (KUST), Kohat 26000, Khyber Pakhtunkhwa, Pakistan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rubina","family":"Nazli","sequence":"additional","affiliation":[{"name":"Institute of Basic Medical Sciences (IBMS), Khyber Medical University (KMU), Peshawar 25100, Khyber Pakhtunkhwa, Pakistan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"May Mohammed","family":"Alrashed","sequence":"additional","affiliation":[{"name":"Chair of Medical and Molecular Genetics Research, Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Saud University, P.O. Box 10219, Riyadh 11433, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hazem K.","family":"Ghneim","sequence":"additional","affiliation":[{"name":"Chair of Medical and Molecular Genetics Research, Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Saud University, P.O. Box 10219, Riyadh 11433, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zia Ul","family":"Haq","sequence":"additional","affiliation":[{"name":"Institute of Public Health & Social Sciences (IPH&SS), Khyber Medical University (KMU), Peshawar 25100, Khyber Pakhtunkhwa, Pakistan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Musarrat","family":"Jabeen","sequence":"additional","affiliation":[{"name":"Department of Gynaecology and Obstetrics, KMU Institute of Medical Sciences, Kohat 26000, Khyber Pakhtunkhwa, Pakistan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sher","family":"Alam Khan","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Genetic Engineering, Kohat University of Science & Technology (KUST), Kohat 26000, Khyber Pakhtunkhwa, Pakistan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ijaz","family":"Ahmad","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Kohat University of Science & Technology (KUST), Kohat 26000, Khyber Pakhtunkhwa, Pakistan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3707-8461","authenticated-orcid":false,"given":"Mohammed","family":"Bourhia","sequence":"additional","affiliation":[{"name":"Laboratory of Chemistry, Biochemistry, Nutrition, and Environment, Faculty of Medicine and Pharmacy, University Hassan II, Casablanca 20000, Morocco"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9395-0563","authenticated-orcid":false,"given":"Mourad A. M.","family":"Aboul-Soud","sequence":"additional","affiliation":[{"name":"Chair of Medical and Molecular Genetics Research, Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Saud University, P.O. Box 10219, Riyadh 11433, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,23]]},"reference":[{"key":"ref_1","first-page":"41","article-title":"Recessive dystrophic epidermolysis bullosa: A review of disease pathogenesis and update on future therapies","volume":"8","author":"Soro","year":"2015","journal-title":"J. Clin. Aesthetic Dermatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1111\/bjd.18921","article-title":"Consensus reclassification of inherited epidermolysis bullosa and other disorders with skin fragility","volume":"183","author":"Has","year":"2020","journal-title":"Br. J. Dermatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"70","DOI":"10.2340\/00015555-2781","article-title":"Epidemiology and Outcome of Squamous Cell Carcinoma in Epidermolysis Bullosa in Australia and New Zealand","volume":"98","author":"Kim","year":"2018","journal-title":"Acta Derm. Venereol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"6","DOI":"10.5582\/irdr.2017.01005","article-title":"Novel and emerging therapies in the treatment of recessive dystrophic epidermolysis bullosa","volume":"6","author":"Rashidghamat","year":"2017","journal-title":"Intractable Rare Dis. Res."},{"key":"ref_5","first-page":"7","article-title":"Best practice guidelines for skin and wound care in epidermolysis bullosa: An international consensus","volume":"1","author":"Denyer","year":"2017","journal-title":"Wounds Int."},{"key":"ref_6","unstructured":"Liy-Wong, C.A.A., Aujla, N., Briggs, L., Campbell, F., El Hamouly, A., and Section of Dermatology at the Hospital for Sick (2021). Epidermolysis Bullosa, a Handbook for EB Patients and Families, DEBRA."},{"key":"ref_7","unstructured":"Adam, M.P., Ardinger, H.H., Pagon, R.A., Wallace, S.E., Bean, L.J.H., Gripp, K.W., Mirzaa, G.M., and Amemiya, A. (1993). Dystrophic Epidermolysis Bullosa. GeneReviews(\u00ae), University of Washington."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1038\/s41576-020-0255-7","article-title":"Exploring human genomic diversity with gnomAD","volume":"21","author":"Koch","year":"2020","journal-title":"Nat. Rev. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.1007\/s00439-020-02199-3","article-title":"The Human Gene Mutation Database (HGMD(\u00ae)): Optimizing its use in a clinical diagnostic or research setting","volume":"139","author":"Stenson","year":"2020","journal-title":"Hum. Genet."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"W99","DOI":"10.1093\/nar\/gkz411","article-title":"Simple ClinVar: An interactive web server to explore and retrieve gene and disease variants aggregated in ClinVar database","volume":"47","author":"Gramm","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1016\/j.ejmg.2017.11.015","article-title":"Pakistan Genetic Mutation Database (PGMD); A centralized Pakistani mutome data source","volume":"61","author":"Qasim","year":"2018","journal-title":"Eur. J. Med. Genet."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1136\/jmg.2006.045302","article-title":"Epidermolysis bullosa. II. Type VII collagen mutations and phenotype-genotype correlations in the dystrophic subtypes","volume":"44","author":"Varki","year":"2007","journal-title":"J. Med. Genet."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1006\/geno.1994.1238","article-title":"Premature termination codons in the type VII collagen gene (COL7A1) underlie severe, mutilating recessive dystrophic epidermolysis bullosa","volume":"21","author":"Christiano","year":"1994","journal-title":"Genomics"},{"key":"ref_14","first-page":"547","article-title":"Injury- and inflammation-driven skin fibrosis: The paradigm of epidermolysis bullosa","volume":"68\u201369","year":"2018","journal-title":"Matrix Bio."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1016\/j.det.2009.12.003","article-title":"Molecular testing in epidermolysis bullosa","volume":"28","author":"Castiglia","year":"2010","journal-title":"Dermatol. Clin."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1669","DOI":"10.1172\/JCI34292","article-title":"A hypomorphic mouse model of dystrophic epidermolysis bullosa reveals mechanisms of disease and response to fibroblast therapy","volume":"118","author":"Fritsch","year":"2008","journal-title":"J. Clin. Investig."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.det.2009.10.020","article-title":"Dystrophic epidermolysis bullosa: Pathogenesis and clinical features","volume":"28","year":"2010","journal-title":"Dermatol. Clin."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"eaas9668","DOI":"10.1126\/scitranslmed.aas9668","article-title":"APOBEC mutation drives early-onset squamous cell carcinomas in recessive dystrophic epidermolysis bullosa","volume":"10","author":"Cho","year":"2018","journal-title":"Sci. Transl. Med."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Bonamonte, D., Filoni, A., De Marco, A., Lospalluti, L., Nacchiero, E., Ronghi, V., Colagrande, A., Giudice, G., and Cazzato, G. (2022). Squamous Cell Carcinoma in Patients with Inherited Epidermolysis Bullosa: Review of Current Literature. Cells, 11.","DOI":"10.3390\/cells11081365"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.clindermatol.2011.03.012","article-title":"Inherited epidermolysis bullosa: New diagnostic criteria and classification","volume":"30","author":"Intong","year":"2012","journal-title":"Clin. Dermatol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Filoni, A., Cicco, G., Cazzato, G., Bosco, A., Lospalluti, L., Tucci, M., Cimmino, A., Foti, C., Marzullo, A., and Bonamonte, D. (2022). Immune Disregulation in Cutaneous Squamous Cell Carcinoma of Patients with Recessive Dystrophic Epidermolysis Bullosa: A Single Pilot Study. Life, 12.","DOI":"10.3390\/life12020213"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1113\/EP086044","article-title":"Gene editing for skin diseases: Designer nucleases as tools for gene therapy of skin fragility disorders","volume":"103","author":"March","year":"2018","journal-title":"Exp. Physiol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"March, O.P., Kocher, T., and Koller, U. (2020). Context-Dependent Strategies for Enhanced Genome Editing of Genodermatoses. Cells, 9.","DOI":"10.3390\/cells9010112"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/j.omtm.2017.06.008","article-title":"Traceless Targeting and Isolation of Gene-Edited Immortalized Keratinocytes from Epidermolysis Bullosa Simplex Patients","volume":"6","author":"Aushev","year":"2017","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1699","DOI":"10.1016\/j.jid.2019.03.1146","article-title":"Gene Editing\u2013Mediated Disruption of Epidermolytic Ichthyosis\u2013Associated KRT10 Alleles Restores Filament Stability in Keratinocytes","volume":"139","author":"March","year":"2019","journal-title":"J. Investig. Dermatol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"5676","DOI":"10.1073\/pnas.1512028113","article-title":"Site-specific genome editing for correction of induced pluripotent stem cells derived from dominant dystrophic epidermolysis bullosa","volume":"113","author":"Shinkuma","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e307","DOI":"10.1038\/mtna.2016.19","article-title":"Gene Editing for the Efficient Correction of a Recurrent COL7A1 Mutation in Recessive Dystrophic Epidermolysis Bullosa Keratinocytes","volume":"5","author":"Chamorro","year":"2016","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.omtn.2018.01.009","article-title":"Deletion of a Pathogenic Mutation-Containing Exon of COL7A1 Allows Clonal Gene Editing Correction of RDEB Patient Epidermal Stem Cells","volume":"11","author":"Chamorro","year":"2018","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1711","DOI":"10.1016\/j.jid.2019.02.015","article-title":"Efficient Gene Reframing Therapy for Recessive Dystrophic Epidermolysis Bullosa with CRISPR\/Cas9","volume":"139","author":"Takashima","year":"2019","journal-title":"J. Investig. Dermatol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"986","DOI":"10.1016\/j.ymthe.2019.03.007","article-title":"Clinically Relevant Correction of Recessive Dystrophic Epidermolysis Bullosa by Dual sgRNA CRISPR\/Cas9-Mediated Gene Editing","volume":"27","author":"Bonafont","year":"2019","journal-title":"Mol. Ther."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1016\/j.molcel.2018.11.031","article-title":"Target-Specific Precision of CRISPR-Mediated Genome Editing","volume":"73","author":"Chakrabarti","year":"2019","journal-title":"Mol. Cell"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"646","DOI":"10.1038\/s41586-018-0686-x","article-title":"Predictable and precise template-free CRISPR editing of pathogenic variants","volume":"563","author":"Shen","year":"2018","journal-title":"Nature"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1985","DOI":"10.1016\/j.jid.2020.02.012","article-title":"Predictable CRISPR\/Cas9-Mediated COL7A1 Reframing for Dystrophic Epidermolysis Bullosa","volume":"140","author":"Kocher","year":"2020","journal-title":"J. Investig. Dermatol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Mayr, E., Ablinger, M., Lettner, T., Murauer, E.M., Guttmann-Gruber, C., Pi\u00f1\u00f3n Hofbauer, J., Hainzl, S., Kaiser, M., Klausegger, A., and Bauer, J.W. (2022). 5\u2019RNA Trans-Splicing Repair of COL7A1 Mutant Transcripts in Epidermolysis Bullosa. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23031732"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1103","DOI":"10.1016\/j.jaad.2014.01.903","article-title":"Inherited epidermolysis bullosa: Updated recommendations on diagnosis and classification","volume":"70","author":"Fine","year":"2014","journal-title":"J. Am. Acad. Dermatol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1016\/j.jaad.2009.03.052","article-title":"Extracutaneous manifestations and complications of inherited epidermolysis bullosa: Part I. Epithelial associated tissues","volume":"61","author":"Fine","year":"2009","journal-title":"J. Am. Acad. Dermatol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1046\/j.1365-2133.2002.04607.x","article-title":"The clinical spectrum of dystrophic epidermolysis bullosa","volume":"146","author":"Horn","year":"2002","journal-title":"Br. J. Dermatol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1136\/bjo.83.3.323","article-title":"The eye in epidermolysis bullosa","volume":"83","author":"Tong","year":"1999","journal-title":"Br. J. Ophthalmol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1097\/ICO.0000000000001525","article-title":"Ophthalmologic Approach in Epidermolysis Bullosa: A Cross-Sectional Study With Phenotype-Genotype Correlations","volume":"37","author":"Mellado","year":"2018","journal-title":"Cornea"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e379","DOI":"10.1038\/mtna.2016.87","article-title":"Antisense Oligonucleotide-mediated Exon Skipping as a Systemic Therapeutic Approach for Recessive Dystrophic Epidermolysis Bullosa","volume":"5","author":"Bremer","year":"2016","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1097","DOI":"10.12688\/f1000research.14974.1","article-title":"Recent advances in understanding and managing epidermolysis bullosa","volume":"7","author":"Kiritsi","year":"2018","journal-title":"F1000Research"}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/12\/7\/1525\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:38:05Z","timestamp":1760139485000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/12\/7\/1525"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,23]]},"references-count":41,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["diagnostics12071525"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics12071525","relation":{},"ISSN":["2075-4418"],"issn-type":[{"type":"electronic","value":"2075-4418"}],"subject":[],"published":{"date-parts":[[2022,6,23]]}}}