Evaluation of signal transduction pathways after transient cutaneous adenoviral gene delivery.
BACKGROUND
Adenoviral vectors have provided effective methods for in vivo gene delivery in therapeutic applications. However, these vectors can induce immune responses that may severely affect the ability of vector re-application. There is limited information about the mechanisms and signal transduction pathways involved in adenoviral recognition. For optimization of cutaneous gene therapy it is necessary to investigate molecular mechanisms of virus recognition in epidermal cells. The aim of this study was to investigate the signal transduction of the innate immunity after adenoviral DNA internalization in keratinocytes.


METHODS
In vitro, keratinocytes were transfected with DNA, in the presence and absence of inhibitors for signalling molecules. In vivo, immunocompetent and athymic mice (n = 3 per group) were twice transduced with an Ad-vector.


RESULTS
The results show an acute induction of type-I-interferon after in vitro transfection. Inhibition of PI3K, p38 MAPK, JNK and NFkappaB resulted in a decreased expression of type-I-interferon. In contrast to immunocompetent mice, athymic mice demonstrated a constant transgene expression and reduced inflammatory response in vivo.


CONCLUSIONS
The results suggest an induction of the innate immunity triggered by cytoplasm localised DNA which is mediated by PI3K-, p38 MAPK-, JNK-, NFkappaB-, JAK/STAT- and ERK1/2-dependent pathways. A stable transgene expression and a reduced inflammatory response in immunodeficient mice have been observed. These results provide potential for an effective adenoviral gene delivery into immunosupressed skin.
Laboratory for Molecular Oncology and Wound Healing, Department of Plastic Surgery, Operative Reference Centre for Soft Tissue Sarcomas, BG University Hospital Bergmannsheil, Ruhr University Bochum, Bochum, Germany. lars.steinstraesser@rub.de
Lars
L
Steinstraesser
Michael
M
Sorkin
Frank
F
Jacobsen
Sammy
S
Al-Benna
Marco Rainer
MR
Kesting
Andreas David
AD
Niederbichler
Jan-Michel
JM
Otte
Tobias
T
Hirsch
Jadwiga
J
Stupka
Hans-Ulrich
HU
Steinau
Matthias
M
Schulte
D000256:Adenoviridae / Q000235:genetics*; D000328:Adult; D000818:Animals; D004247:DNA / Q000378:metabolism; D004705:Endocytosis; D018014:Gene Transfer Techniques; D006801:Humans; D007113:Immunity, Innate / Q000276:immunology; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics*; D012867:Skin / Q000378:metabolism*; D051193:Toll-Like Receptors / Q000378:metabolism; D019076:Transgenes / Q000235:genetics; D055815:Young Adult
D012333:RNA, Messenger; D051193:Toll-Like Receptors; D004247:DNA
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