All Title Author
Keywords Abstract

PLOS Genetics  2014 

Molecular Identification of Collagen 17a1 as a Major Genetic Modifier of Laminin Gamma 2 Mutation-Induced Junctional Epidermolysis Bullosa in Mice

DOI: doi/10.1371/journal.pgen.1004068

Full-Text   Cite this paper   Add to My Lib


Epidermolysis Bullosa (EB) encompasses a spectrum of mechanobullous disorders caused by rare mutations that result in structural weakening of the skin and mucous membranes. While gene mutated and types of mutations present are broadly predictive of the range of disease to be expected, a remarkable amount of phenotypic variability remains unaccounted for in all but the most deleterious cases. This unexplained variance raises the possibility of genetic modifier effects. We tested this hypothesis using a mouse model that recapitulates a non-Herlitz form of junctional EB (JEB) owing to the hypomorphic jeb allele of laminin gamma 2 (Lamc2). By varying normally asymptomatic background genetics, we document the potent impact of genetic modifiers on the strength of dermal-epidermal adhesion and on the clinical severity of JEB in the context of the Lamc2jeb mutation. Through an unbiased genetic approach involving a combination of QTL mapping and positional cloning, we demonstrate that Col17a1 is a strong genetic modifier of the non-Herlitz JEB that develops in Lamc2jeb mice. This modifier is defined by variations in 1–3 neighboring amino acids in the non-collagenous 4 domain of the collagen XVII protein. These allelic variants alter the strength of dermal-epidermal adhesion in the context of the Lamc2jeb mutation and, consequentially, broadly impact the clinical severity of JEB. Overall the results provide an explanation for how normally innocuous allelic variants can act epistatically with a disease causing mutation to impact the severity of a rare, heritable mechanobullous disorder.


[1]  Fine JD (2010) Inherited epidermolysis bullosa. Orphanet J Rare Dis 5: 12. doi: 10.1186/1750-1172-5-12
[2]  Sawamura D, Nakano H, Matsuzaki Y (2010) Overview of epidermolysis bullosa. J Dermatol 37: 214–219. doi: 10.1111/j.1346-8138.2009.00800.x
[3]  Uitto J, McGrath JA, Rodeck U, Bruckner-Tuderman L, Robinson EC (2010) Progress in epidermolysis bullosa research: toward treatment and cure. J Invest Dermatol 130: 1778–1784. doi: 10.1038/jid.2010.90
[4]  Bruckner-Tuderman L, Has C (2012) Molecular heterogeneity of blistering disorders: the paradigm of epidermolysis bullosa. J Invest Dermatol 132: E2–5. doi: 10.1038/skinbio.2012.2
[5]  Karamatic Crew V, Burton N, Kagan A, Green CA, Levene C, et al. (2004) CD151, the first member of the tetraspanin (TM4) superfamily detected on erythrocytes, is essential for the correct assembly of human basement membranes in kidney and skin. Blood 104: 2217–2223. doi: 10.1182/blood-2004-04-1512
[6]  Fine JD, Eady RA, Bauer EA, Bauer JW, Bruckner-Tuderman L, et al. (2008) The classification of inherited epidermolysis bullosa (EB): Report of the Third International Consensus Meeting on Diagnosis and Classification of EB. J Am Acad Dermatol 58: 931–950. doi: 10.1016/j.jaad.2008.02.004
[7]  Petronius D, Bergman R, Ben Izhak O, Leiba R, Sprecher E (2003) A comparative study of immunohistochemistry and electron microscopy used in the diagnosis of epidermolysis bullosa. Am J Dermatopathol 25: 198–203. doi: 10.1097/00000372-200306000-00003
[8]  Uitto J, Pulkkinen L, McLean WH (1997) Epidermolysis bullosa: a spectrum of clinical phenotypes explained by molecular heterogeneity. Mol Med Today 3: 457–465. doi: 10.1016/s1357-4310(97)01112-x
[9]  Bruckner-Tuderman L (2008) Dystrophic epidermolysis bullosa: pathogenesis and clinical features. Dermatol Clin 28: 107–114. doi: 10.1016/j.det.2009.10.020
[10]  Ussar S, Wang HV, Linder S, Fassler R, Moser M (2006) The Kindlins: subcellular localization and expression during murine development. Exp Cell Res 312: 3142–3151. doi: 10.1016/j.yexcr.2006.06.030
[11]  Aumailley M, Rousselle P (1999) Laminins of the dermo-epidermal junction. Matrix Biol 18: 19–28. doi: 10.1016/s0945-053x(98)00004-3
[12]  Aumailley M, Bruckner-Tuderman L, Carter WG, Deutzmann R, Edgar D, et al. (2005) A simplified laminin nomenclature. Matrix Biol 24: 326–332. doi: 10.1016/j.matbio.2005.05.006
[13]  Kiritsi D, Kern JS, Schumann H, Kohlhase J, Has C, et al. (2011) Molecular mechanisms of phenotypic variability in junctional epidermolysis bullosa. J Med Genet 48: 450–457. doi: 10.1136/jmg.2010.086751
[14]  Varki R, Sadowski S, Pfendner E, Uitto J (2006) Epidermolysis bullosa. I. Molecular genetics of the junctional and hemidesmosomal variants. J Med Genet 43: 641–652. doi: 10.1136/jmg.2005.039685
[15]  Dang N, Klingberg S, Rubin AI, Edwards M, Borelli S, et al. (2008) Differential expression of pyloric atresia in junctional epidermolysis bullosa with ITGB4 mutations suggests that pyloric atresia is due to factors other than the mutations and not predictive of a poor outcome: three novel mutations and a review of the literature. Acta Derm Venereol 88: 438–448. doi: 10.2340/00015555-0484
[16]  Titeux M, Pendaries V, Tonasso L, Decha A, Bodemer C, et al. (2008) A frequent functional SNP in the MMP1 promoter is associated with higher disease severity in recessive dystrophic epidermolysis bullosa. Hum Mutat 29: 267–276. doi: 10.1002/humu.20647
[17]  Schneider H, Muhle C, Pacho F (2007) Biological function of laminin-5 and pathogenic impact of its deficiency. Eur J Cell Biol 86: 701–717. doi: 10.1016/j.ejcb.2006.07.004
[18]  Muhle C, Jiang QJ, Charlesworth A, Bruckner-Tuderman L, Meneguzzi G, et al. (2005) Novel and recurrent mutations in the laminin-5 genes causing lethal junctional epidermolysis bullosa: molecular basis and clinical course of Herlitz disease. Hum Genet 116: 33–42. doi: 10.1007/s00439-004-1210-y
[19]  Nakano A, Chao SC, Pulkkinen L, Murrell D, Bruckner-Tuderman L, et al. (2002) Laminin 5 mutations in junctional epidermolysis bullosa: molecular basis of Herlitz vs. non-Herlitz phenotypes. Hum Genet 110: 41–51. doi: 10.1007/s00439-001-0630-1
[20]  Bubier JA, Sproule TJ, Alley LM, Webb CM, Fine JD, et al. (2010) A mouse model of generalized non-Herlitz junctional epidermolysis bullosa. J Invest Dermatol 130: 1819–1828. doi: 10.1038/jid.2010.46
[21]  Sproule TJ, Roopenian DC, Sundberg JP (2012) A direct method to determine the strength of the dermal-epidermal junction in a mouse model for epidermolysis bullosa. Exp Dermatol 21: 453–455. doi: 10.1111/j.1600-0625.2012.01482.x
[22]  Li R, Lyons MA, Wittenburg H, Paigen B, Churchill GA (2005) Combining data from multiple inbred line crosses improves the power and resolution of quantitative trait loci mapping. Genetics 169: 1699–1709. doi: 10.1534/genetics.104.033993
[23]  Broman KW, Wu H, Sen S, Churchill GA (2003) R/qtl: QTL mapping in experimental crosses. Bioinformatics 19: 889–890. doi: 10.1093/bioinformatics/btg112
[24]  Sen S, Satagopan JM, Broman KW, Churchill GA (2007) R/qtlDesign: inbred line cross experimental design. Mamm Genome 18: 87–93. doi: 10.1007/s00335-006-0090-y
[25]  Meng X, Klement JF, Leperi DA, Birk DE, Sasaki T, et al. (2003) Targeted inactivation of murine laminin gamma2-chain gene recapitulates human junctional epidermolysis bullosa. J Invest Dermatol 121: 720–731. doi: 10.1046/j.1523-1747.2003.12515.x
[26]  Bauer JW, Lanschuetzer C (2003) Type XVII collagen gene mutations in junctional epidermolysis bullosa and prospects for gene therapy. Clin Exp Dermatol 28: 53–60. doi: 10.1046/j.1365-2230.2003.01192.x
[27]  Tasanen K, Floeth M, Schumann H, Bruckner-Tuderman L (2000) Hemizygosity for a glycine substitution in collagen XVII: unfolding and degradation of the ectodomain. J Invest Dermatol 115: 207–212. doi: 10.1046/j.1523-1747.2000.00049.x
[28]  Chavanas S, Gache Y, Tadini G, Pulkkinen L, Uitto J, et al. (1997) A homozygous in-frame deletion in the collagenous domain of bullous pemphigoid antigen BP180 (type XVII collagen) causes generalized atrophic benign epidermolysis bullosa. J Invest Dermatol 109: 74–78. doi: 10.1111/1523-1747.ep12276614
[29]  Franzke CW, Tasanen K, Schumann H, Bruckner-Tuderman L (2003) Collagenous transmembrane proteins: collagen XVII as a prototype. Matrix Biol 22: 299–309. doi: 10.1016/s0945-053x(03)00051-9
[30]  Giudice GJ, Emery DJ, Diaz LA (1992) Cloning and primary structural analysis of the bullous pemphigoid autoantigen BP180. J Invest Dermatol 99: 243–250. doi: 10.1111/1523-1747.ep12616580
[31]  Clark LD, Clark RK, Heber-Katz E (1998) A new murine model for mammalian wound repair and regeneration. Clin Immunol Immunopathol 88: 35–45. doi: 10.1006/clin.1998.4519
[32]  Billings T, Parvanov ED, Baker CL, Walker M, Paigen K, et al. (2013) DNA binding specificities of the long zinc-finger recombination protein PRDM9. Genome biology 14: R35. doi: 10.1186/gb-2013-14-4-r35
[33]  Parvanov ED, Petkov PM, Paigen K (2010) Prdm9 controls activation of mammalian recombination hotspots. Science 327: 835. doi: 10.1126/science.1181495
[34]  Rousselle P, Keene DR, Ruggiero F, Champliaud MF, Rest M, et al. (1997) Laminin 5 binds the NC-1 domain of type VII collagen. J Cell Biol 138: 719–728. doi: 10.1083/jcb.138.3.719
[35]  Borradori L, Sonnenberg A (1999) Structure and function of hemidesmosomes: more than simple adhesion complexes. J Invest Dermatol 112: 411–418. doi: 10.1046/j.1523-1747.1999.00546.x
[36]  Sugawara K, Tsuruta D, Ishii M, Jones JC, Kobayashi H (2008) Laminin-332 and -511 in skin. Exp Dermatol 17: 473–480. doi: 10.1111/j.1600-0625.2008.00721.x
[37]  Hirako Y, Usukura J, Nishizawa Y, Owaribe K (1996) Demonstration of the molecular shape of BP180, a 180-kDa bullous pemphigoid antigen and its potential for trimer formation. J Biol Chem 271: 13739–13745. doi: 10.1074/jbc.271.23.13739
[38]  Tasanen K, Tunggal L, Chometon G, Bruckner-Tuderman L, Aumailley M (2004) Keratinocytes from patients lacking collagen XVII display a migratory phenotype. Am J Pathol 164: 2027–2038. doi: 10.1016/s0002-9440(10)63762-5
[39]  Nishie W, Kiritsi D, Nystrom A, Hofmann SC, Bruckner-Tuderman L (2011) Dynamic interactions of epidermal collagen XVII with the extracellular matrix: laminin 332 as a major binding partner. Am J Pathol 179: 829–837. doi: 10.1016/j.ajpath.2011.04.019
[40]  Van den Bergh F, Eliason SL, Giudice GJ (2011) Type XVII collagen (BP180) can function as a cell-matrix adhesion molecule via binding to laminin 332. Matrix Biol 30: 100–108. doi: 10.1016/j.matbio.2010.10.005
[41]  Yuen WY, Pas HH, Sinke RJ, Jonkman MF (2011) Junctional epidermolysis bullosa of late onset explained by mutations in COL17A1. Br J Dermatol 164: 1280–1284. doi: 10.1111/j.1365-2133.2011.10359.x
[42]  Schumann H, Hammami-Hauasli N, Pulkkinen L, Mauviel A, Kuster W, et al. (1997) Three novel homozygous point mutations and a new polymorphism in the COL17A1 gene: relation to biological and clinical phenotypes of junctional epidermolysis bullosa. Am J Hum Genet 60: 1344–1353. doi: 10.1086/515463
[43]  Fu CL, Giudice GJ, Van den Bergh F (2006) Protein structural analysis of BP180 mutant isoforms linked to non-Herlitz junctional epidermolysis bullosa. J Invest Dermatol 126: 232–234. doi: 10.1038/sj.jid.5700024
[44]  Darling TN, Yee C, Bauer JW, Hintner H, Yancey KB (1999) Revertant mosaicism: partial correction of a germ-line mutation in COL17A1 by a frame-restoring mutation. J Clin Invest 103: 1371–1377. doi: 10.1172/jci4338
[45]  Floeth M, Bruckner-Tuderman L (1999) Digenic junctional epidermolysis bullosa: mutations in COL17A1 and LAMB3 genes. Am J Hum Genet 65: 1530–1537. doi: 10.1086/302672
[46]  Ludwig RJ, Muller S, Marques A, Recke A, Schmidt E, et al. (2012) Identification of quantitative trait loci in experimental epidermolysis bullosa acquisita. J Invest Dermatol 132: 1409–1415. doi: 10.1038/jid.2011.466
[47]  Huggins KW, Camarota LM, Howles PN, Hui DY (2003) Pancreatic triglyceride lipase deficiency minimally affects dietary fat absorption but dramatically decreases dietary cholesterol absorption in mice. J Biol Chem 278: 42899–42905. doi: 10.1074/jbc.m303422200
[48]  Doerge RW, Churchill GA (1996) Permutation tests for multiple loci affecting a quantitative character. Genetics 142: 285–294.
[49]  Lander E, Kruglyak L (1995) Genetic dissection of complex traits: guidelines for interpreting and reporting linkage results. Nature genetics 11: 241–247. doi: 10.1038/ng1195-241


comments powered by Disqus