title: Complementation of the DNA repair deficiency in human xeroderma pigmentosum group a and C cells by recombinant adenovirus-mediated gene transfer.
abstract: Nucleotide excision repair (NER) is one of the most versatile DNA repair mechanisms, ensuring the proper functioning and trustworthy transmission of genetic information in all living cells. The phenotypic consequences caused by NER defects in humans are autosomal recessive diseases such as xeroderma pigmentosum (XP). This syndrome is the most sun-sensitive disorder leading to a high frequency of skin cancer. The majority of patients with XP carry mutations in the XPA or XPC genes that encode proteins involved in recognition of DNA damage induced by UV light at the beginning of the NER process. Cells cultured from XPA and XPC patients are hypersensitive to UV light, as a result of malfunctioning DNA repair. So far there is no effective long-term treatment for these patients. Skin cancer prevention can only be achieved by strict avoidance of sunlight exposure or by the use of sunscreen agents. We have constructed recombinant adenoviruses carrying the XPA and XPC genes that were used to infect XP-A and XP-C immortalized and primary fibroblast cell lines. UV survival curves and unscheduled DNA synthesis confirmed complete phenotypic reversion in XP DNA repair deficient cells with no trace of cytotoxicity. Moreover, transgene expression is stable for at least 60 days after infection. This efficient adenovirus gene delivery approach may be an important tool to better understand XP deficiency and the causes of DNA damage induced skin cancer.
authors.affiliation: Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Alysson Renato
authors.initials: AR
authors.lastname: Muotri
authors.forename: Maria Carolina Nasser
authors.initials: MC
authors.lastname: Marchetto
authors.forename: Luiz Fernando Celidonio
authors.initials: LF
authors.lastname: Zerbini
authors.forename: Towia A
authors.initials: TA
authors.lastname: Libermann
authors.forename: Armando Morais
authors.initials: AM
authors.lastname: Ventura
authors.forename: Alain
authors.initials: A
authors.lastname: Sarasin
authors.forename: Carlos Frederico Martins
authors.initials: CF
authors.lastname: Menck
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics*; D002461:Cell Line, Transformed; D004249:DNA Damage; D004260:DNA Repair / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000502:physiology; D005260:Female; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology / Q000528:radiation effects; D005816:Genetic Complementation Test; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008297:Male; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000502:physiology; D014466:Ultraviolet Rays / Q000009:adverse effects; D014983:Xeroderma Pigmentosum / Q000145:classification / Q000235:genetics / Q000473:pathology*; D051760:Xeroderma Pigmentosum Group A Protein
chemical_list: D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C497314:XPA protein, human; D051760:Xeroderma Pigmentosum Group A Protein