Cutis laxa(CL)isaninheritedoracquiredconnective

tissuedisordercharacterizedbyredundant,loose,
sagging,andinelasticskinthatisduetoanabnormal
deficiencyofdermalelasticfibersorreducedelastin
synthesisoffibroblasts.

AutosomaldominantCLisusuallyamildcutaneous
disease,sometimesaccompaniedbygastrointestinal
diverticuli,hernias,orgenitalprolapse.Pulmonaryartery
stenosis,aortic,andarterialdilatationandtortuosity,
Raynaud'sphenomenon,bronchiectasis,and
emphysemaarerarecomplications.Twoprevious
studieshaveshownmutationsintheelastingene(ELN)
inADCLpatients.
 Methodology

DNAisolation,direct
Patientsamplesand sequenceanalysis,
SouthernandRNA
celllineswere andisolationofthe
blotanalyses
obtained duplication
breakpoint

Mutant-specific
Metaboliclabeling, SMARTRACEand
antibodyand
immunoprecipitation, cloningofthemutant
immunofluorescent
andwesternblotting mRNA
staining
 Results
Histological and electron microscopic evaluation of dermal elastic fiber

A. Hematoxylineosin staining (100um)

B-C. Elastic van Giesson (EVG) staining (100um)
D-E. Transmission Electron Microscopy (2um)
 Results
ical characterization of familycutis laxa(CL)-1
(A) Prematurelyredundantandsaggingfacial
skinoftheprobandIII.1atage40y.(B)
ThepedigreeofCL-1indicatesautosomal
dominantinheritance.
(B) Hatchedsymbolsindicateindividuals(I.2,
II.9,andIII.11)whowerereportedly
affectedbutwerenotavailablefor
examination.
(C) Smalldiamonds(IV.2andIV.3)indicate
miscarriages;largediamondsdenote
multipleunaffectedindividualswithinthe
samegeneration.
(D) (C)Posterioanterior(PA)chestX-ray
(viewedinAPposition)ofIII.1shortly
beforelungtransplantationattheageof
51y.
(E) Hyperexpandedlungfieldsindicate
emphysema.Bilateralbasalshadows
reveallungfibrosis.Notetheenlargedleft
pulmonaryarteryalongtheleftheart
borderdemonstratingpulmonary
hypertension.
(F) (D)InelasticskinandgeneralizedCLin
individualIV.4atbirth.
Results
Immunofluorescence localization
of the mutant tropoelastin (TE)
in dermal fibroblasts
Subconfluentculturescontrol(A)andcutis
laxa(CL)patientIII.1(B)fibroblastswerefixed
andpermeabilizedforintracellularlocalizationof
themutantTEproteinusingtheintron10
antibody.Whereasonlybackgroundstaining
wasseeninthecontrolcells(A),aprominent,
punctate,peri-nuclearstainingwasseeninthe
patientfibroblasts(B).Post-confluentfibroblast
culturesfromcontrol(C)andCL-1patientIII.1
(D)werefixedandstainedwiththeintron10
antibody(specificforthemutantprotein).
Controlswerenegative(C),whereasthe
extracellularmatrixofCLcellswaspositivefor
themutantprotein(D).Nucleiofcontrolcells
werecounterstainedtodemonstratethe
presenceofthecelllayerbecausestainingwith
theintron10antibodywasnegative.Post-
confluentfibroblastculturesfromcontrol(E)and
CL-1patientIII.1(F)werefixedandstainedwith
ananti-TEantibody(recognizingbothnormal
andmutantproteins).Theabundanceand
intensityofelasticfibersweresimilarinboth
patientandcontrolcultures.Toindicatecell
density,nucleiwerealsostained.
 Conclusion
This study demonstrated that a partial tandem duplication in ELN causes
autosomal dominant CL with severe chronic obstructive pulmonary disease
(COPD). The following lines of evidence support the conclusion that this
duplication is the disease-causing defect: (1) both affected individuals
studied in the family carried the same duplication; (2) the duplication was
absent in 136 normal control individuals; (3) the gene defect resulted in a
major change in the reading frame and in the synthesis of mutant TE with
partial ECM incorporation and partial intracellular retention of the abnormal
protein; and (4) tissue samples from CL patients with the ELN duplication
containedabnormalelasticfibersascharacterizedbyalackofassociationof
theamorphouselastinwithmicrofibrillarcomponentsofthefiber.

Elastin mutations can cause CL associated with a severe pulmonary

phenotype.SynthesisofabnormalTEmayinterferewithelasticfiberfunction
throughadominant-negativeoragainoffunctionmechanism.