Functional Re-expression of Laminin-5 in Laminin-γ2-deficient Human Keratinocytes Modifies Cell Morphology, Motility, and Adhesion

Gagnoux‐Palacios, Laurent; Vailly, Joëlle; Durand-Clément, Monique; Wagner, Ernst; Ortonne, Jean‐Paul; Meneguzzi, Guerríno

doi:10.1074/jbc.271.31.18437

Cited by 72 publications

(53 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Indeed, hemidesmosomal structures are not observed in artificial epithelia reconstituted with H-JEB keratinocytes that either produce no laminin-5, or, like LSV5 cells, secrete scant amounts of this adhesion ligand. 17,20 In this system, the hemidesmosomes reconstitute in correspondence to the anchoring fibrils of the de-epidermized dermis. Therefore, as in normal skin, the laminin-5 deposited on the basement membrane induces formation of the anchoring filaments and mediates their connection with the collagen VII molecules on the basement membrane.…”

Section: Discussionmentioning

confidence: 99%

“…In a previous work, we described the re-expression of laminin-5 in the immortalized H-JEB keratinocytes LSV5-R infected with a retrovirus expressing a wild-type laminin ␥2 chain. 20 The transduced cells restructured focal adhesions, modified their morphology and motility, and improved their adhesion to the culture substrate, but failed to synthesize hemidesmosomal structures. These results contrasted with the observation that immortalized JEB keratinocytes deficient in the expression of integrin ␣6␤4 infected with retroviruses expressing the adequate wild-type transgene synthesize hemidesmosomal-like structures in vitro.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Corrective gene transfer of keratinocytes from patients with junctional epidermolysis bullosa restores assembly of hemidesmosomes in reconstructed epithelia

et al. 1998

View full text Add to dashboard Cite

Herlitz junctional epidermolysis bullosa (H-JEB) providesdeposited into the extracellular matrix. Re-expression of a promising model for somatic gene therapy of heritable laminin-5 induced cell spreading, nucleation of hemidesmechano-bullous disorders. This genodermatosis is mosomal-like structures and enhanced adhesion to the culcaused by the lack of laminin-5 that results in absence of ture substrate. Organotypic cultures performed with the hemidesmosomes (HD) and defective adhesion of squamtransduced keratinocytes, reconstituted epidermis closely ous epithelia. To establish whether re-expression of lamiadhering to the mesenchyme and presenting mature heminin-5 can restore assembly of the dermal-epidermal attachdesmosomes, bridging the cytoplasmic intermediate filament structures lacking in the H-JEB skin, we corrected the ments of the basal cells to the anchoring filaments of the genetic mutation hindering expression of the ␤3 chain of basement membrane. Our results provide the first evilaminin-5 in human H-JEB keratinocytes by transfer of a dence of phenotypic reversion of JEB keratinocytes by laminin ␤3 transgene. The transduced keratinocytes synsomatic gene therapy and demonstrate that genetic treatthesized a recombinant ␤3 polypeptide that assembled ment of the mild forms of skin blistering diseases and other with the endogenous laminin ␣3 and ␥2 chains into a bioinherited extracellular matrix pathologies is a realistic goal. logically active laminin-5 that was secreted, processed and

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Corrective gene transfer of keratinocytes from patients with junctional epidermolysis bullosa restores assembly of hemidesmosomes in reconstructed epithelia

et al. 1998

View full text Add to dashboard Cite

show abstract

“…Prior approaches restored expression of a number of genes to epidermal cells in JEB and other skin diseases in vitro 24,25 and in regenerated tissue in vivo; 2,21-23 however, they relied on amphotropic retroviruses with all their attendant drawbacks. In the plasmid-based approach presented here, functional SB transposase is required for sustained corrective gene delivery because a nonfunctional mutant failed to maintain gene transfer, even in vitro.…”

Section: Discussionmentioning

confidence: 99%

Sustainable correction of junctional epidermolysis bullosa via transposon-mediated nonviral gene transfer

et al. 2003

View full text Add to dashboard Cite

“…We recently reported that retroviral vector-mediated transduction of laminin-5 cDNA restores synthesis of functional laminin-5 and induces changes in focal adhesion and motility in Herlitz JEB keratinocytes (28). This study is the first step for an application of in vivo gene transfer for the treatment of the JEB genodermatosis.…”

Section: Fig 5 Sequence Of the 27-kbmentioning

confidence: 99%

“…NIH-3T3 fibroblasts (American Type Culture Collection, CRL 1658) were grown in Dulbecco's modified Eagle's medium supplemented with 10% bovine serum. Plasmid DNAs were purified using the silica columns from Qiagen (Hylden, Germany), and transfections were carried out in 24-well dishes using LipofectAMINE (Life Technologies, Inc.) as detailed elsewhere (28). Each DNA construct was tested in triplicate wells in three separate experiments.…”

Section: Methodsmentioning

confidence: 99%

Murine Laminin α3A and α3B Isoform Chains Are Generated by Usage of Two Promoters and Alternative Splicing

Ferrigno¹,

Virolle²,

Galliano³

et al. 1997

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

We already identified two distinct laminin ␣3A and ␣3B chain isoforms which differ in their amino-terminal ends and display different tissue-specific expression patterns. In this study we have investigated whether these two different isoforms are products of the same laminin ␣3 (lama3) gene and transcribed from one or two separate promoters. Genomic clones were isolated that encompass the sequences upstream to the 5 ends of both the ␣3A and the ␣3B cDNAs. Sequence analysis of the region upstream to the ␣3A open reading frame revealed the presence of a TATA box and potential binding sites for responsive elements. By primer extension analysis, the transcription start site of the ␣3B mRNA isoform was defined. The sequences upstream to the ␣3B mRNA transcription start site do not contain a TATA box near the transcription initiation sites, but AP-1, AP-2, and Sp1 consensus binding site sequences were identified. The genomic regions located immediately upstream of the ␣3A and ␣3B transcription start sites were shown to possess promoter activities in transfection experiments. In the promoter regions, response elements for the acute phase reactant signal and NF-interleukin 6 were found, and their possible relevance in the context of inflammation and wound healing is discussed. Our results demonstrate that the lama3 gene produces the two polypeptides by alternative splicing and contains two promoters, which regulate the production of the two isoforms ␣3A and ␣3B.

show abstract

Functional Re-expression of Laminin-5 in Laminin-γ2-deficient Human Keratinocytes Modifies Cell Morphology, Motility, and Adhesion

Abstract: Herlitz junctional epidermolysis bullosa (H-JEB

Cited by 72 publications

References 43 publications

Corrective gene transfer of keratinocytes from patients with junctional epidermolysis bullosa restores assembly of hemidesmosomes in reconstructed epithelia

Corrective gene transfer of keratinocytes from patients with junctional epidermolysis bullosa restores assembly of hemidesmosomes in reconstructed epithelia

Sustainable correction of junctional epidermolysis bullosa via transposon-mediated nonviral gene transfer

Murine Laminin α3A and α3B Isoform Chains Are Generated by Usage of Two Promoters and Alternative Splicing

Contact Info

Product

Resources

About