Novel Homozygous and Compound Heterozygous COL17A1 Mutations Associated with Junctional Epidermolysis Bullosa Michaela Floeth, Heike Schäcke, Nadja Hammami-Hauasli,

Slides:

Advertisements

Similar presentations

Lipodermatosclerosis is Characterized by Elevated Expression and Activation of Matrix Metalloproteinases: Implications for Venous Ulcer Formation Yared.

Advertisements

Epidermolysis Bullosa: Novel and De Novo Premature Termination Codon and Deletion Mutations in the Plectin Gene Predict Late-Onset Muscular Dystrophy

The Missense Mutation p

Skin-Specific Expression of ank-393, a Novel Ankyrin-3 Splice Variant

Herlitz Junctional Epidermolysis Bullosa: Novel and Recurrent Mutations in the LAMB3 Gene and the Population Carrier Frequency Aoi Nakano, Ellen Pfendner,

Transient Bullous Dermolysis of the Newborn Associated with Compound Heterozygosity for Recessive and Dominant COL7A1 Mutations Nadja Hammami-Hauasli,

The Autoantigen in Anti-p200 Pemphigoid Is Synthesized by Keratinocytes and Fibroblasts and Is Distinct from Nidogen-2 Silke C. Hofmann, Ursula Voith,

BM-40(Osteonectin, SPARC) Is Expressed Both in the Epidermal and in the Dermal Compartment of Adult Human Skin Nicholas Hunzelmann, Martin Hafner, Sabine.

Deletion of the Major Bullous Pemphigoid Epitope Region of Collagen XVII Induces Blistering, Autoimmunization, and Itching in Mice Tiina Hurskainen,

Molecular Basis of Kindler Syndrome in Italy: Novel and Recurrent Alu/Alu Recombination, Splice Site, Nonsense, and Frameshift Mutations in the KIND1.

Combination of a Novel Frameshift Mutation (1929delCA) and a Recurrent Nonsense Mutation (W610X) of the LAMB3 Gene in a Japanese Patient with Herlitz.

Volume 88, Issue 4, Pages (February 1997)

A Novel Component of Epidermal Cell–Matrix and Cell–Cell Contacts: Transmembrane Protein Type XIII Collagen Sirkku Peltonen, Maria Hentula, Pasi Hägg,

Differential Expression of a Novel Gene in Response to hsp27 and Cell Differentiation in Human Keratinocytes Mojgan Hell-Pourmojib, Peter Neuner, Robert.

Correction of Dog Dystrophic Epidermolysis Bullosa by Transplantation of Genetically Modified Epidermal Autografts Yannick Gache, Didier Pin, Laurent.

Molecular Mechanisms of Junctional Epidermolysis Bullosa: Col15 Domain Mutations Decrease the Thermal Stability of Collagen XVII Laura Väisänen, Cristina.

Generation of a Functional Non-Shedding Collagen XVII Mouse Model: Relevance of Collagen XVII Shedding in Wound Healing Joanna Jacków, Andreas Schlosser,

Natural Gene Therapy May Occur in All Patients with Generalized Non-Herlitz Junctional Epidermolysis Bullosa with COL17A1 Mutations Anna M.G. Pasmooij,

Anna M. G. Pasmooij, Hendri H. Pas, Franciska C. L

Marcel F. Jonkman, Anna M. G. Pasmooij, Suzanne G. M. A

Novel Mutations in the LAMC2 Gene in Non-Herlitz Junctional Epidermolysis Bullosa: Effects on Laminin-5 Assembly, Secretion, and Deposition Daniele Castiglia,

Targeted Inactivation of Murine Laminin γ2-Chain Gene Recapitulates Human Junctional Epidermolysis Bullosa Xianmin Meng, John F. Klement, Dominic A.

Colocalization of Kindlin-1, Kindlin-2, and Migfilin at Keratinocyte Focal Adhesion and Relevance to the Pathophysiology of Kindler Syndrome J.E. Lai-Cheong,

Marie-Thérèse Leccia Journal of Investigative Dermatology

Epidermolysis Bullosa: Novel and De Novo Premature Termination Codon and Deletion Mutations in the Plectin Gene Predict Late-Onset Muscular Dystrophy

Mild Recessive Bullous Congenital Ichthyosiform Erythroderma due to a Previously Unidentified Homozygous Keratin 10 Nonsense Mutation Akiko Tsubota,

Anna M. G. Pasmooij, Hendri H. Pas, Franciska C. L

Epidermolysis Bullosa Simplex Associated with Pyloric Atresia Is a Novel Clinical Subtype Caused by Mutations in the Plectin Gene (PLEC1) Hiroyuki Nakamura,

Novel SLC39A4 Mutations in Acrodermatitis Enteropathica

Comparative Mutation Detection Screening of the Type VII Collagen Gene (COL7A1) Using the Protein Truncation Test, Fluorescent Chemical Cleavage of Mismatch,

Cycloheximide Facilitates the Identification of Aberrant Transcripts Resulting from a Novel Splice-Site Mutation in COL17A1 in a Patient with Generalized.

Laminin-5 Mutational Analysis in an Italian Cohort of Patients with Junctional Epidermolysis Bullosa Patrizia Posteraro, Naomi De Luca, Guerrino Meneguzzi,

Identification and Sequencing of a Putative Variant of Proopiomelanocortin in Human Epidermis and Epidermal Cells in Culture Gong Can, Zalfa Abdel-Malek,

A Homozygous Nonsense Mutation in Type XVII Collagen Gene (COL17A1) Uncovers an Alternatively Spliced mRNA Accounting for an Unusually Mild Form of Non-Herlitz.

Michaela Floeth, Leena Bruckner-Tuderman

Inherited Junctional Epidermolysis Bullosa in the German Pointer: Establishment of a Large Animal Model Annabelle Capt, Flavia Spirito, Eric Guaguere,

Rapid Decay of α6 Integrin Caused by a Mis-Sense Mutation in the Propeller Domain Results in Severe Junctional Epidermolysis Bullosa with Pyloric Atresia

P16INK4a and p14ARF Tumor Suppressor Genes Are Commonly Inactivated in Cutaneous Squamous Cell Carcinoma Victoria L. Brown, Catherine A. Harwood, Tim.

Maternal Uniparental Meroisodisomy in the LAMB3 Region of Chromosome 1 Results in Lethal Junctional Epidermolysis Bullosa Yasuko Takizawa, Leena Pulkkinen,

Compound Heterozygous TGM1 Mutations Including a Novel Missense Mutation L204Q in a Mild Form of Lamellar Ichthyosis Masashi Akiyama, Itsuro Matsuo

A Novel Homozygous Mutation Affecting Integrin α6 in a Case of Junctional Epidermolysis Bullosa with Pyloric Atresia Detected In Utero by Ultrasound Examination

Deletion of the Cytoplasmatic Domain of BP180/Collagen XVII Causes a Phenotype with Predominant Features of Epidermolysis Bullosa Simplex Marcel Huber,

A Novel Point Mutation Affecting the Tyrosine Kinase Domain of the TRKA Gene in a Family with Congenital Insensitivity to Pain with Anhidrosis Shinichi.

A Deep-Intronic FERMT1 Mutation Causes Kindler Syndrome: An Explanation for Genetically Unsolved Cases Nadja Chmel, Sorina Danescu, Amelie Gruler, Dimitra.

Compound Heterozygosity for Non-Sense and Mis-Sense Mutations in Desmoplakin Underlies Skin Fragility/Woolly Hair Syndrome Neil V. Whittock, Hong Wan,

Compound Heterozygosity for a Recessive Glycine Substitution and a Splice Site Mutation in the COL7A1 Gene Causes an Unusually Mild Form of Localized.

Revertant Mosaicism in Recessive Dystrophic Epidermolysis Bullosa

Gabriella Esposito, Giuseppe Rescigno, Francesco Salvatore

Compound Heterozygosity for Novel Splice Site Mutations in the BPAG2/COL17A1 Gene Underlies Generalized Atrophic Benign Epidermolysis Bullosa Leena Pulkkinen,

A Novel Gene Expressed in Human Keratinocytes with Long-Term In Vitro Growth Potential is Required for Cell Growth Laure Aurelian, Cynthia C. Smith,

A Usual Frameshift and Delayed Termination Codon Mutation in Keratin 5 Causes a Novel Type of Epidermolysis Bullosa Simplex with Migratory Circinate Erythema

Jittima Dhitavat, Leonard Dode, Natalie Leslie, Anavaj Sakuntabhai

Identification of a Lethal Form of Epidermolysis Bullosa Simplex Associated with a Homozygous Genetic Mutation in Plectin Maryse Bonduelle, Linda De.

Expression of FcRn, the MHC Class I-Related Receptor for IgG, in Human Keratinocytes Karla Cauza, Gabriele Hinterhuber, Ruth Dingelmaier-Hovorka, Karin.

Complete Maternal Isodisomy of Chromosome 3 in a Child with Recessive Dystrophic Epidermolysis Bullosa but No Other Phenotypic Abnormalities Hiva Fassihi,

Genotype–Phenotype Correlation in Recessive Dystrophic Epidermolysis Bullosa: When Missense Doesn't Make Sense Vesarat Wessagowit, Soo-Chan Kim, Se Woong.

The 97 kDa Linear IgA Bullous Dermatosis Antigen is not Expressed in a Patient with Generalized Atrophic Benign Epidermolysis Bullosa with a Novel Homozygous.

Autoantibodies to BP180 Associated with Bullous Pemphigoid Release Interleukin-6 and Interleukin-8 from Cultured Human Keratinocytes Enno Schmidt, Stanislaus.

Loss of Cell Adhesion in Dsg3bal–Pas Mice with Homozygous Deletion Mutation (2079del14) in the Desmoglein 3 Gene Leena Pulkkinen, Yoo Won Choi, Anisha.

Frances J.D. Smith, W.H. Irwin McLean

Neil V. Whittock, Frances J. Smith, W.H. Irwin McLean

Gonosomal Mosaicism for a Nonsense Mutation (R1947X) in the NF1 Gene in Segmental Neurofibromatosis Type 1 Claudia Consoli, Celia Moss, Stuart Green,

Ellen Pfendner, Jouni Uitto Journal of Investigative Dermatology

Novel Keratin 14 Mutations in Patients with Severe Recessive Epidermolysis Bullosa Simplex Cristina Has, Yow-Ren Chang, Andreas Volz, Doris Hoeping,

Is Screening of the Candidate Gene Necessary in Unrelated Partners of Members of Families with Herlitz Junctional Epidermolysis Bullosa? Alfred Klausegger,

Marcel F. Jonkman, Prof. Dr, Hendri H

Kit-Sing Au, Adelaide A. Hebert, E. Steve Roach, Hope Northrup

Exon Skipping in IVD RNA Processing in Isovaleric Acidemia Caused by Point Mutations in the Coding Region of the IVD Gene Jerry Vockley, Peter K. Rogan,

Novel Mutations in the LAMB3 Gene Shared by Two Japanese Unrelated Families with Herlitz Junctional Epidermolysis Bullosa, and Their Application for Prenatal.

Identification of Novel pro-α2(IX) Collagen Gene Mutations in Two Families with Distinctive Oligo-Epiphyseal Forms of Multiple Epiphyseal Dysplasia Paul.

Presentation transcript:

Novel Homozygous and Compound Heterozygous COL17A1 Mutations Associated with Junctional Epidermolysis Bullosa Michaela Floeth, Heike Schäcke, Nadja Hammami-Hauasli, Leena Bruckner-Tuderman Journal of Investigative Dermatology Volume 111, Issue 3, Pages 528-533 (September 1998) DOI: 10.1046/j.1523-1747.1998.00325.x Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Pedigrees of the probands and mutations in the COL17A1 gene. Pedigrees demonstrate recessive inheritance of the mutations. (A) family 1; (B) family 2; (C) family 3; (D) family 4. (A) Heteroduplex bands were detected on CSGE gels of PCR product spanning exon 8 (middle panel). Control DNA (lane 1) and the patient’s DNA (lane 2) showed homoduplex bands when tested alone. The faster migration of the patient’s homoduplex band suggested a deletion in this fragment. Heteroduplex bands were seen in mixed samples of the patient’s and control DNA (lane 3). Didesoxynucleotide sequencing of the patient’s DNA revealed a homozygous 2 bp deletion in exon 8, designated 520delAG (right panel), whereas the deletion was absent from a control (left panel). (B) CSGE analysis of PCR product spanning exon 43 (middle panel) revealed a homoduplex band in control DNA (lane 1) and the patient’s DNA (lane 2), but heteroduplex bands when the samples were mixed with each other (lane 3). Direct didesoxynucleotide sequencing of the patients’ PCR products revealed a homozygous 1 bp deletion of nucleotide G in exon 43, designated 2965delG (right panel), which was absent from a control (left panel). (C) CSGE analysis of the PCR products spanning exons 18 and 37 revealed heteroduplex bands with the patient’s DNA, indicating compound heterozygosity of the proband (middle panels). The upper middle panel shows CSGE analysis of PCR product spanning exon 37 (paternal mutation); control DNA (lane 1) showed a homoduplex band and the patient’s DNA (lane 2) homo- and heteroduplex bands. The lower homoduplex band (asterisk) was excised, eluted, reamplified, and subjected to didesoxynucleotide sequencing that revealed a heterozygous 2 bp deletion in exon 37, designated 2666delTT (right panel), which was absent from a control (left panel). This heterozygous deletion was also found in the father’s DNA. The lower middle panel shows CSGE analysis of PCR product spanning exon 18 (maternal mutation); control DNA (lane 1) showed a homoduplex band and the patient’s DNA (lane 2) heteroduplex bands. Direct didesoxynucleotide sequencing of the patients’ PCR products revealed a heterozygous 1721G→A transition in exon 18, designated G539E (right panel), which was absent from a control (left panel). This heterozygous mutation was also found in the mother’s DNA. In the pedigree, the black areas indicate the paternal and the dotted areas the maternal allele. (D) Identification of the paternal mutation. CSGE analysis of PCR product spanning exon 51 (middle panel) revealed a homoduplex band in control (lane 1) and the mother’s DNA (lane 4). In contrast, heteroduplex bands were seen with the patient’s (lane 2), the father’s (lane 3), and the sister’s DNA (lane 5). Direct didesoxynucleotide sequencing of the patient’s PCR products revealed a heterozygous 3781C→T transition in exon 51, designated R1226X (right panel). The mutation was also found in the unaffected father and sister, but not in a control or the mother (left panel). Extensive mutation screening did not reveal other changes in the COL17A1 gene and therefore the maternal mutation remained elusive. Journal of Investigative Dermatology 1998 111, 528-533DOI: (10.1046/j.1523-1747.1998.00325.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Northern blotting of collagen XVII mRNA from normal and GABEB keratinocytes. A strong signal of the 6 kb collagen XVII mRNA was found in control keratinocytes (Co). In contrast, collagen XVII mRNA isolated from the patient 1’s keratinocytes (Pat 1) produced a very weak signal, suggesting nonsense-mediated mRNA decay in these cells. The panels were intentionally overexposed to demonstrate the presence of the weak collagen XVII mRNA band in the patient’s cells. A digoxigenin-labeled GAPDH cDNA was used as a reference probe to correct for differences in RNA loading. Journal of Investigative Dermatology 1998 111, 528-533DOI: (10.1046/j.1523-1747.1998.00325.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 IF staining of the patients’ skin with antibodies to cutaneous basement membrane components. The biopsies and antibodies used were: (a) patient 1, 1A8C; (b) patient 1, laminin 5; (c) patient 2, 1A8C; (d) patient 2, integrin α6; (e) patient 3, 1A8C; (f) patient 3, collagen VII; (g) patient 4, 1A8C; (h) patient 4, collagen VII. In (g), a weak reactivity with the antibody 1A8C was observed in the basal keratinocytes of the detached epidermis. The stainings with antibodies to other basement membrane proteins were normal. In blistered skin of patient 2, antibodies to integrin α6 localized to the blister roof (d). Antibodies to collagen VII stained the blister base in separated skin of patients 3 (f) and 4 (h). The staining patterns were consistent with junctional blistering. Scale bar: 40 μm. Journal of Investigative Dermatology 1998 111, 528-533DOI: (10.1046/j.1523-1747.1998.00325.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Immunodetection of collagen XVII in keratinocytes in vitro. (a–d) IF staining of GABEB keratinocytes with domain-specific antibodies to collagen XVII. In control cells, the antibody 1D1 to the extracellular domain showed a positive reaction in the rough endoplasmatic reticulum and in the cell periphery, corresponding to the transmembrane localization (a). The antibody produced a negative reaction in the keratinocytes of patient 1 (b). Staining of keratinocytes of patient 3 with the antibody 1A8C to the intracellular domain revealed a positive diffuse intracellular fluorescence (c), and with the antibody 1D1 to the extracellular domain a diffuse weak reaction (d). Scale bar: 10 μm. (e) Immunoblotting of keratinocyte extracts with polyclonal antibodies to the NC16a domain. Normal controls contained the 180 kDa α1(XVII)-chain (lanes 1, 3, 5). In contrast, cell extracts of patient 1 (lane 2), patient 3 (lane 4), and patient 4 (lane 6) lacked the band, indicating absence of stable collagen XVII in these cells. The 65 kDa band in lane 6 was occasionally seen with the human autoantiserum used, but its identity has not been characterized. The migration positions of molecular weight markers are indicated on the right. Journal of Investigative Dermatology 1998 111, 528-533DOI: (10.1046/j.1523-1747.1998.00325.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions