Generation and analysis of conditional inducible transgene mice for the analysis of cFLIP in the skin
Final Report Abstract
Control of the cell death is of critical importance for regulation of cell and tissue homeostasis. Key molecules in apoptotic signalling such as FADD, caspase-8 and cFLIP were shown to play a critical role in the embryonal development. Moreover, skin specific, constitutive deletion of FADD or caspase-8 resulted in inflammatory skin disease. The goal of this project was to investigate the significance of cFLIP in the skin homeostasis. We have used Cre/LoxP mediated recombination system to generated skin specific cFLIP knock out mouse model. Our data demonstrated that in contrast to FADD and caspase-8, constitutive deletion of cFLIP in skin resulted in embryonic lethality, indicating the relevance of cFLIP for skin development. To analyze the impact of cFLIP deletion in adult animals we next generated mouse model with skin-specific inducible postnatal deletion of cFLIP. Acute deletion of cFLIP resulted in severe macroscopically detectable inflammation and epidermal loss in the skin. To gain more insights in the mechanism of cell death in cFLIP deficient keratinocytes in vivo we have analyzed in details the cell death in cFLIP deficient primary keratinocytes. Our in vitro analysis revealed that TNF-mediated autocrine loop in the skin has a critical importance for apoptosis in cFLIP-deficient keratinocytes. The crutial, functional role of TNF for epidermal cell death was further confirmed by our in vitro and in vivo inhibition experiments using TNF-R2-Fc fusion protein. We have shown that epidermal cFLIP protein was lost in patients with severe drug reactions associated with epidermal apoptosis. Our data demonstrate the importance of cFLIP for the integrity of the epidermis and for silencing of spontaneous skin inflammation. Taken together, our findings provide insights into the regulation of the cell death machinery of the epidermis and warrant future studies to dissect the impact of loss of cFLIP in a number of inflammatory and neoplastic diseases of the skin.
Publications
- (2013) cFLIP regulates skin homeostasis and protects against TNF-induced keratinocyte apoptosis. Cell Rep. 31;5(2)
Panayotova-Dimitrova D, Feoktistova M, Ploesser M, Kellert B, Hupe M, Horn S, Makarov R, Jensen F, Porubsky S, Schmieder A, Zenclussen AC, Marx A, Kerstan A, Geserick P, He YW, Leverkus M
(See online at https://doi.org/10.1016/j.celrep.2013.09.035) - (2013) Thymidine analogues suppress autophagy and adipogenesis in cultured adipocytes. Antimicrob Agents Chemother. 57(1):543-51
Stankov MV, Panayotova-Dimitrova D, Leverkus M, Schmidt RE, Behrens GM
(See online at https://doi.org/10.1128/AAC.01560-12) - (2014) Histone deacetylase inhibitors induce apoptosis in myeloid leukemia by suppressing autophagy. Leukemia 28(3)
Stankov MV, El Khatib M, Kumar Thakur B, Heitmann K, Panayotova-Dimitrova D, Schoening J, Bourquin JP, Schweitzer N, Leverkus M, Welte K, Reinhardt D, Li Z, Orkin SH, Behrens GM, Klusmann JH
(See online at https://doi.org/10.1038/leu.2013.264) - RIPping the Skin Apart: Necroptosis Signaling in Toxic Epidermal Necrolysis. (2015) Invest Dermatol.,135(8)
Panayotova-Dimitrova D, Feoktistova M, Leverkus M.J
(See online at https://doi.org/10.1038/jid.2015.159) - (2017) RIPK1 Suppresses a TRAF2-Dependent Pathway to Liver Cancer. Cancer Cell. Jan 9;31(1)
Schneider AT, Gautheron J, Feoktistova M, Roderburg C, Loosen SH, Roy S, Benz F, Schemmer P, Büchler MW, Nachbur U, Neumann UP, Tolba R, Luedde M, Zucman-Rossi J, Panayotova-Dimitrova D, Leverkus M, Preisinger C, Tacke F, Trautwein C, Longerich T, Vucur M, Luedde T
(See online at https://doi.org/10.1016/j.ccell.2016.11.009)
