Protein therapeutics for junctional epidermolysis bullosa: incorporation of recombinant beta3 chain into laminin 332 in beta3-/- keratinocytes in vitro.

[junctional epidermolysis bullosa]

Junctional epidermolysis bullosa (JEB ) is an inherited mechanobullous disease characterized by reduced adherence of the epidermal keratinocytes to the underlying dermis , and is often caused by the absence of functional laminin 332 due to the lack or dysfunction of its beta 3 chain . As there are no specific therapies for JEB , we tested whether a protein replacement strategy could be applicable for the restoration of the laminin 332 assembly and reversion of the JEB phenotype in human keratinocytes that lack beta 3 subunit . Here , we developed the protocol for production and purification of the biologically active recombinant beta 3 chain . Next , we demonstrated that delivery of recombinant beta 3 polypeptide into the endoplasmic reticulum of the immortalized beta 3 -null keratinocytes led to the restoration of the laminin 332 assembly , secretion , and deposition into the basement membrane zone , as confirmed by Western blot analysis , confocal immunofluorescent microscopy in vitro , and on cultured organotypic human JEB skin reconstructs . Although the amount of laminin 332 produced by protein-treated beta 3 -null keratinocytes is lower than that in normal human keratinocytes , our results demonstrate the applicability of the recombinant proteins for JEB treatment and open new perspectives for the development of novel therapeutics for this inherited , currently intractable , skin disorder .

Diseases
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Diseases presenting "specific therapies" symptom

alpha-thalassemia

junctional epidermolysis bullosa

lymphangioleiomyomatosis

primary hyperoxaluria type 1

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