S Hadj-Rabia et al. Acrogeria as a Novel Laminopathy
CONFLICT OF INTEREST The authors state no conflict of interest.
ACKNOWLEDGMENTS This study was funded by R01-AR0056755 from the National Institute of Arthritis Musculoskeletal and Skin Diseases, a grant from Valeant Pharmaceuticals for the PEER study, and a grant from the Breast Cancer Research Foundation (Nathanson) for informatics analysis.
David J. Margolis1,2,5, Jayanta Gupta1,5, Andrea J. Apter3, Ole Hoffstad1, Maryte Papadopoulos1, Tim R. Rebbeck1, Bradley Wubbenhorst4 and Nandita Mitra1 1
Department of Biostatistics and Epidemiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA; 2 Department of Dermatology, Perelman School of Medicine, University of Pennsylvania, Philadelphia Pennsylvania, USA; 3Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia Pennsylvania, USA and 4Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia Pennsylvania, USA E-mail:
[email protected] 5 These two authors share first authorship.
REFERENCES Brown SJ, McLean WH (2012) One remarkable molecule: filaggrin. J Invest Dermatol 132: 751–62 Bussman C, Weidinger S, Novak N (2011) Genetics of atopic dermatitis. J German Soc Dermatol 9:670–9 Ellinghaus D, Baurecht H, Esparza-Gordillo J et al. (2013) High-density genotyping study indentifies four new susceptibility loci for atopic dermatitis. Nat Genet 45:808–12
persistent atopic dermatitis in African American subjects. J Allergy Clin Immunol 133:784–9 Pellerin L, Henry J, Hsu CY et al. (2013) Defects in filaggrin-like proteins in both lesional and nonlesional atopic skin. J Allergy Clin Immunol 131:1094–102 Perusquia-Ortiz AM, Oji V, Sauerland MC et al. (2013) Complete filaggrin deficiency in ichthyosis vulgaris is associated with only moderate changes in epidermal permeability barrier function profile. J Eur Acad Dermatol Venereol 27:1552–8
Fleckman P, Brumbaugh S (2002) Absence of granular layer and keratohylin define a morphologically distinct subset of individuals with ichthyosis vulgaris. Exper Dermatol 11:327–36
Thaswer-Esmail F, Jakasa I, Todd G et al. (2014) South African amaXhosa patient with atopic dermatitis have decreased levels of filaggrin breakdown products but no loss-of-function mutations in filaggrin. J Allergy Clin Immunol 133:280–2
Henry J, Toulza E, Hsu CY et al. (2012) Update on the epidermal differentiation complex. Front Biosci 17:1517–32
The 1000 Genomes Project Consortium (2012) An integrated map of genetic variation from 1,092 human genomes. Nature 491:56–65
Marenholz I, Rivera VA, Esparza-Gordillo J et al. (2011) Association screening in the Epidermal Differentiation Complex (EDC) identifies an SPRR3 repeat number variant as a risk factor for eczema. J Investig Dermatol 131:1644–9
Thyssen JP, Godoy-Gijon E, Elias PM (2013) Ichthyosis vulgaris: the filaggrin mutation disease. Br J Dermatol 168:155–8
Margolis DJ, Apter AJ, Gupta J et al. (2012) The persistence of atopic dermatitis and Filaggrin mutations in a US longitudinal cohort. J Allergy Clin Immunol 130:912–7 Margolis DJ, Gupta J, Apter AJ et al. (2014) Filaggrin-2 variation is associated wtih more
Winge MC, Bilcha KD, Lieden A et al. (2011a) Novel filaggrin mutation but no other loss-of-function variants found in Ethiopian patients with atopic dermatitis. Br J Dermatol 165:1074–80 Winge MC, Bilcha KD, Lieden A et al. (2011b) Novel filaggrin mutation but no other loss-of-function variants found in Ethiopian patients with atopic dermatitis. Br J Dermatol 165:1074–80
A New Lamin A Mutation Associated with Acrogeria Syndrome Journal of Investigative Dermatology (2014) 134, 2274–2277; doi:10.1038/jid.2014.158; published online 24 April 2014
TO THE EDITOR Acrogeria (OMIM#201200), first described in 1940 (Gottron, 1940), is a form of skin atrophy combined with mottled hyperpigmentation and subcutaneous tissue atrophy giving an aged appearance. The pathogenesis of acrogeria seems heterogeneous, as abnormal biosynthesis of type III collagen is suspected in some cases, making Gottron-type acrogeria and vasculartype Ehlers–Danlos syndrome (vEDS, OMIM#130050) allelic diseases (Pope et al., 1996; Jansen et al., 2000; Hashimoto et al., 2004). Several clinical features of acrogeria recall the dermatologic defects observed
in Hutchinson–Gilford progeria syndrome (HGPS OMIM#176670), mandibuloacral dysplasia type A (MADAOMIM#248370), or mandibuloacral dysplasia type B (MADB, OMIM #608612) phenotypes, known to be caused by Lamins A/C defects, due to mutations either in the LMNA gene that encodes them or in the ZMPSTE24 gene, which encodes a protease involved in Lamin A processing (Novelli et al., 2002; De SandreGiovannoli et al., 2003; Eriksson et al., 2003; Ahmad et al., 2010). Lamins are nuclear intermediate filaments that are involved in nuclear architecture and functions including chromatin organization or DNA replication, transcription,
Abbreviations: HGPS, Hutchinson–Gilford progeria syndrome; MAD, mandibuloacral dysplasia; RT-PCR, reverse transcription–PCR; vEDS, vascular-type Ehlers–Danlos syndrome Accepted article preview online 1 April 2014; published online 24 April 2014
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and repair, reviewed in Prokocimer et al., 2009. The patient affected with acrogeria was born in 1969 from a healthy mother and a probably affected father, who was reported by the patient to have the same skin aspect and had a mitral and aortic valve replacement. The patient has a brother and a son who are not affected. At the age of 36 years, he was referred to our department for ‘‘prematurely aged’’ appearance. He presented with a thin nose and translucent skin on the face (Figure 1a, A). The distal portion of the lower limbs seemed lipodystrophic and the patient presented short clavicles (Figure 1a, B). Translucent skin of the arms and abdomen had pigmentation abnormalities (Figure 1a, B, C). The hands and the feet showed atrophic dermis and loss of subcutaneous fat (Figure 1a, B, D). The hands had a very
S Hadj-Rabia et al. Acrogeria as a Novel Laminopathy
aged appearance with prominent veins and osteolyses of the distal phalanges (Figure 1a, D). All these clinical signs are strikingly evocative of HGPS or MAD phenotypes. Cardiovascular, pulmonary, ophthalmological, and central nervous system examinations revealed no abnormalities (data not shown). Liver and renal functions were normal, and the patient was not diabetic. The patient consented to his image being used. A skin biopsy of the right arm showed slight epidermal hyperplasia (Figure 1a, E), irregular and fragmented elastic fibers (orcein staining, Figure 1a, F), and disorganization of collagen bundles with degenerative aspect of collagen network (Figure 1a, G). EDTA blood was obtained from the patient and
a
A
C
family members following written consent, in adherence to the Declaration of Helsinki principles. We screened LMNA gene at the genomic level and observed the c.1771T4A heterozygous transition in exon 11 (Figure 1b). The mutation was observed only in the proband and was absent in the rest of the unaffected family. Unfortunately, the DNA of the affected father was not available because he died several years earlier (Figure 1c). At the protein level, the identified mutation was predicted to replace a Cysteine with a Serine at amino acid position 591 (p.Cys591Ser), in the C-terminal part specific to lamin A (from 567–664), being absent from lamin C (stop at 566). This mutation was absent in more than 100 healthy internal controls, as well as in 1500
b LMNA, c.1771T>A; p.Cys591Ser G G G AC C T GC GG GC A
D
Reference sequence G G G AC C N GC GG GC A
Patient sequence
B E
c F
G c.1771T>A/+
+/+
+/+ Figure 1. Clinical features and molecular characterization. (a) Clinical feature (36 years old). (A) Patient’s face, (B) body, (C) abdomen with pigmented macules, and (D) aged appearance of the hands. Prominent veins (dotted arrow) and wrinkles (black arrow) are indicated. (E) Hematoxylin and eosin staining, scale bar ¼ 2 mm. Thinned dermis with sweat glands ascents (asterisk). (F) Hematoxylin and eosin staining, scale bar ¼ 4 mm. Disorganization of collagen bundles with degenerative aspect is shown by asterisk. (G) Orcein staining, scale bar ¼ 4 mm. Fragmentation of the elastic fibers is indicated by asterisk. (b) Genetic analysis of the LMNA gene in the proband (bottom) compared with control (top). Heterozygous mutation, T to A transition, underlined by red lines, is positioned at the coding nucleotide 1771 in exon 11 (c.1771T4A; p.Cys591Ser). (c) Pedigree of the family. The affected individual is shown by arrowhead, unfilled symbols correspond to unaffected subjects, and individual for whom the disease is suspected is shown as a gray symbol. Diagonal slash indicates the deceased subject. Images used with the patient’s consent.
other patients tested in our molecular diagnostic laboratory in other clinical contexts evocative of a laminopathy; it is not reported in the UMD-LMNA database (http://www.umd.be/LMNA/) nor in the 1000 genome resources (http://browser.1000genomes.org/index. html) or the UCSC genome browser (http://genome.ucsc.edu/) as a SNP. On the basis of deleted Lamin A isoforms associated with LMNA exon 11 mutations, we performed cDNA explorations with overlapping and ‘‘fulllength’’ primers but did not observe any shorter transcripts (data not shown). In concordance with the reverse transcription–PCR (RT-PCR) analyses, Western blots on primary dermal fibroblasts did not reveal any truncated form at the protein level and showed normal Lamins A/C amounts (Figure 2a, lane 1) in comparison with healthy control (lane 2). No band was evidenced with a progerin-specific antibody, opposite to a HGPS patient, used as a progerin positive ‘‘control’’ (lane 3; Figure 2a). In parallel, as the vEDS and acrogeria are often misdiagnosed, and because a mutation in the COL3A1 gene has already been reported in a patient with Gottron-type Acrogeria (Jansen et al., 2000), we excluded COL3A1 mutations or transcript alterations by RT-PCR and direct sequencing from the COL3A1 cDNA issued from the patient’s fibroblast cell line (data not shown). Several laminopathies are characterized by nuclear abnormalities, reflecting a nuclear fragility caused by nuclear lamina instability (Schreiber and Kennedy, 2013). To support the pathogenicity of the identified mutation, we performed indirect immunofluorescence experiments using anti-lamins A/C on patient and control fibroblasts and evidenced that 35.2% of patient’s nuclei presented a misshaped nuclear structure characterized by blebs (Figure 2b, asterisk), but also abundant micronuclei representing 8% of total nuclei (Figure 2b, arrowhead), compared with 11 and 1.3%, respectively, in control fibroblasts (Supplementary Figure S1 online). Antibodies directed against Emerin and Lamin B2, both Lamin A partners, confirmed these anomalies. Staining with NuMA antibody showed a heterogeneous nuclear staining in www.jidonline.org 2275
S Hadj-Rabia et al. Acrogeria as a Novel Laminopathy
1
2
3
M Lamin A Progerin Lamin C
GAPDH
Patient
Lamin A/C
Control
*
* Emerin
Lamin B2
*
NuMA
Figure 2. Functional analyses. (a) Western blot. Merged pictures of western blots using anti-lamins A/C (red), anti-progerin (green), and anti-glyceraldehyde-3phosphate dehydrogenase (GAPDH; green), as a loading control. Patient’s proteins are loaded in lane 1, the healthy control’s proteins in lane 2, and the Hutchinson–Gilford progeria syndrome patient’s proteins in lane 3. Progerin band is present only for the Hutchinson–Gilford progeria syndrome patient (middle band, lane 3). (b) Indirect immunofluorescence. Analyses on skin fibroblast cultures of patient (A–H) compared with control (A’–H’). Lamins A/C (A, A’), emerin (B, B’), and lamin B2 (C, C’) antibodies are in green. All these antibodies evidenced blebs indicated by asterisks and micronuclei by arrowheads. NuMA antibody (D, D’) is in red and shows a heterogeneous and mottled distribution pattern in patient (D) compared with control (D’). All images were counterstained with DAPI, in order to evidence DNA (E–H, E’–H’). Scale bars ¼ 20 mm.
‘‘clumps’’ in our patient (Figure 2b, D) compared with the homogeneous punctuate staining observed in the control (Figure 2b, D’). Despite the fact that only one patient is described in this study, which is a frequent situation in extremely rare disorders as are some laminopathies, the segregation of the disease in the family, the absence of the mutation in large control populations and databases, the presence of nuclear abnormalities, and the fact that the same Cysteine residue is mutated, with a different substitution (p.Cys591Phe) in a patient affected with partial lipodystrophy, insulin resistance, aortic stenosis, and
hypertrophic cardiomyopathy (Arau´joVilar et al., 2008), all are very strong arguments in favor of its pathogenicity. It is well established indeed that in the context of Lamins A/C mutations, different changes of the same amino acid can give rise to different clinical phenotypes (Bonne et al., 1999). The case we report points to the involvement of the LMNA gene in ‘‘acrogeria’’, including this disease in the clinical spectrum of progeroid laminopathies. On the basis of the cardiac disease and dermatological manifestations observed in the father, another possible interpretation is that both cases,
2276 Journal of Investigative Dermatology (2014), Volume 134
the father and the son, are affected with a form of atypical progeria syndrome with major dermatologic features found in this patient (Doubaj et al., 2012; Kane et al., 2013). In both cases, the patient we report, sharing p.Cys591Ser in Lamin A protein and characterized by nuclear abnormalities in fibroblast culture, extends the clinical spectrum of progeroid laminopathies and suggests that the LMNA gene should be screened in patients presenting with segmental progeroid traits, as well as in acrogeria patients presenting wild-type COL3A1 sequences. However, further functional studies will be needed to determine the
T Goldsmith et al. Heterogeneity in p63 Syndromes
mutant function more precisely, and additional patient recruitment would be very helpful. CONFLICT OF INTEREST The authors state no conflict of interest.
ACKNOWLEDGMENTS
Mole´culaire, Centre de Compe´tences Maladies Vasculaires Rares, CHU de Montpellier, Montpellier, France and 7Laboratoire de Cytologie Clinique et Cytoge´ne´tique, UF Ge´ne´tique Me´dicale, CHU de Nıˆmes, Nıˆmes, France E-mail:
[email protected] or
[email protected] 8 These authors contributed equally to this work.
We are grateful to the patient who consented to his image being used.
Smail Hadj-Rabia1,2,8, Jacob Mashiah1,2,8, Patrice Roll3,4, Amandine Boyer5, Patrice Bourgeois5, Philippe Khau Van Kien6,7, Nicolas Le´vy3,5, Annachiara De Sandre-Giovannoli3,5, Christine Bodemer1,2 and Claire Navarro3
SUPPLEMENTARY MATERIAL
1
Arau´jo-Vilar D, Lado-Abeal J, Palos-Paz F et al. (2008) A novel phenotypic expression associated with a new mutation in LMNA gene, characterized by partial lipodystrophy, insulin resistance, aortic stenosis and hypertrophic cardiomyopathy. Clin Endocrinol (Oxf) 69:61–8
Department of Dermatology, Reference Center for Genodermatoses and Rare Skin Diseases (MAGEC), INSERM U781, Universite´ Paris Descartes—Sorbonne Paris Cite´, Paris, France; 2 Institut Imagine, Hoˆpital Universitaire NeckerEnfants Malades, Assistance Publique-Hoˆpitaux de Paris, Paris, France; 3Aix Marseille Universite´, GMGF, INSERM, UMR_S 910, Marseille, France; 4Laboratoire de Biologie Cellulaire, Assistance Publique-Hoˆpitaux de Marseille, Hoˆpital d’Enfants La Timone, Marseille, France; 5Laboratoire de Ge´ne´tique Mole´culaire, Assistance Publique-Hoˆpitaux de Marseille, Hoˆpital d’Enfants La Timone, Marseille, France; 6Laboratoire de Ge´ne´tique
atypical progeria syndrome. Am J Med Genet A 158A:2881s–7s Eriksson M, Brown WT, Gordon LB et al. (2003) Recurrent de novo point mutations in lamin A cause Hutchinson-Gilford progeria syndrome. Nature 423:293–8 Gottron H (1940) Familia¨re Akrogerie. Arch Dermatol Syph 181:571 Hashimoto C, Abe M, Onozawa N et al. (2004) Acrogeria (Gottron type): a vascular disorder? Br J Dermatol 151:497–501
Supplementary material is linked to the online version of the paper at http://www.nature.com/jid
Jansen T, de Paepe A, Luytinck N et al. (2000) COL3A1 mutation leading to acrogeria (Gottron Type). Br J Dermatol 142:178–80
REFERENCES
Kane MS, Lindsay ME, Judge DP et al. (2013) LMNA-associated cardiocutaneous progeria: an inherited autosomal dominant premature aging syndrome with late onset. Am J Med Genet A 161A:1599–611
Ahmad Z, Zackai E, Medne L et al. (2010) Early onset mandibuloacral dysplasia due to compound heterozygous mutations in ZMPSTE24. Am J Med Genet A 152A:2703–10
Bonne G, Di Barletta MR, Varnous S et al. (1999) Mutations in the gene encoding lamin A/C cause autosomal dominant Emery-Dreifuss muscular dystrophy. Nat Genet 21:285–8 De Sandre-Giovannoli A, Bernard R, Cau P et al. (2003) Lamin a truncation in HutchinsonGilford progeria. Science 300:2055 Doubaj Y, De Sandre-Giovannoli A, Vera EV et al. (2012) An inherited LMNA gene mutation in
Novelli G, Muchir A, Sangiuolo F et al. (2002) Mandibuloacral dysplasia is caused by a mutation in LMNA-encoding lamin A/C. Am J Hum Genet 71:426–31 Pope FM, Narcisi P, Nicholls AC et al. (1996) COL3A1 mutations cause variable clinical phenotypes including acrogeria and vascular rupture. Br J Dermatol 135:163–81 Prokocimer M, Davidovich M, Nissim-Rafinia M et al. (2009) Nuclear lamins: key regulators of nuclear structure and activities. J Cell Mol Med 13:1059–85 Schreiber KH, Kennedy BK (2013) When lamins go bad: nuclear structure and disease. Cell 152:1365–75
A Mutation in TP63 Causing a Mild Ectodermal Dysplasia Phenotype Journal of Investigative Dermatology (2014) 134, 2277–2280; doi:10.1038/jid.2014.159; published online 24 April 2014
TO THE EDITOR The transcription factor p63 has been shown to have a pivotal role during ectodermal, orofacial, and limb development (Mills et al., 1999; Yang et al., 1999; Rinne et al., 2007). So far, mutations in TP63 encoding p63 have been linked to three major phenotypes: ectodermal defects, split hand/foot malformation, or orofacial clefting (Celli et al., 1999; van Bokhoven et al., 1999;
Ianakiev et al., 2000; McGrath et al., 2001; Rinne et al., 2007). Ectodermal dysplasia syndromes associated with TP63 mutation are always inherited in an autosomal dominant manner (Koster, 2010) and include the ectrodactyly, ectodermal dysplasia, and cleft lip/ palate syndrome (EEC), the ankyloblepharon–ectodermal defects–cleft lip/ palate syndrome (AEC), the limb– mammary syndrome, the acro–dermato–
Abbreviations: AEC, ankyloblepharon–ectodermal defects–cleft lip/palate syndrome; EEC, ectrodactyly, ectodermal dysplasia, and cleft lip/palate syndrome; RHS, Rapp–Hodgkin syndrome; TI, transcription inhibitory Accepted article preview online 27 March 2014; published online 24 April 2014
ungual–lacrimal–tooth syndrome, and the Rapp–Hodgkin syndrome (RHS; Rinne et al., 2007). Through large-scale mutation screening, several phenotype–genotype correlations have emerged: although mutations altering the p63 DNA–binding domain have been shown to cause EEC, mutations affecting the sterile alphamotif domain of the protein cause mostly AEC (Mills et al., 1999; Yang et al., 1999; McGrath et al., 2001; Lo Iacono et al., 2008; Koster, 2010). In the present study, we identified a mutation affecting a conserved residue of the p63 transcription inhibitory (TI) domain causing a phenotype previously unreported to the best of our knowledge. www.jidonline.org 2277
