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miRNAs as integrative determinants of the keratinocyte response to UVB.

English title miRNAs as integrative determinants of the keratinocyte response to UVB.
Applicant Dotto Gian-Paolo
Number 130576
Funding scheme Sinergia
Research institution Département de Biochimie Faculté de Biologie et Médecine Université de Lausanne
Institution of higher education University of Lausanne - LA
Main discipline Cellular Biology, Cytology
Start/End 01.08.2010 - 30.09.2013
Approved amount 1'600'000.00
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Keywords (10)

microRNAs ; ultraviolet light; skin; keratinocyte; cancer; Notch; Nrf2; PPAR; in vivo delivery; transgenic mice

Lay Summary (English)

Lead
Lay summary
The skin is an organ of great clinical relevance. It also provides an excellent model system to understand integrated control of tissue homeostasis under normal conditions, in response to external insults (most notably UV light), in the wound healing process, during aging, and in tumorigenesis. In concert with protein-encoding genes, small non-coding RNA molecules, called microRNAs (miRNAs) play a key role in coordinating the overall gene expression response of cells and tissues to changing environmental conditions. The main goal of this proposal is to explore the role that miRNAs play in the UV response of keratinocytes. More specifically, we will test the hypothesis that miRNAs control the acute UV response of keratinocytes, impinging on intrinsic regulation of these cells and their modulation of other cell types, in a possible cross-talk with three specific cell regulatory pathways, involving the p53/Notch, Nrf2 and PPAR?/? transcription factors. We will further test the hypothesis that miRNAs play an essential function in the long-term UV response of keratinocytes, in particular in skin cancer development.By a combination of biochemical and functional assays, we will assess the role of specific miRNAs in UV-mediated cell cycle arrest, differentiation, apoptosis, inflammation as well as long term tumor formation. Novel strategies for in vivo delivery of miRNAs/antagomiRs to the skin will be developed. Putative targets of relevant miRNAs will be identified by combined bioinformatic and biochemical approaches, followed by in vitro and in vivo validation. Finally, cross-regulation between specific miRNAs and the p53/Notch1, PPAR?/?, and Nrf2 pathways will be determined.These ambitious goals will be achieved by a close collaboration between three groups with strong and complementary expertise in skin biology, and a fourth group with strong expertise in miRNA biology/technology. The project is expected to shed light onto the mechanisms involved in the UV response of the skin and to identify key regulators involved in skin protection/repair after UV-induced damage. By a combined comparative approach of mouse and human systems, the work is likely to produce significant novel insights with strong implications for human health. In addition, this project will strengthen the fields of skin biology/ miRNA biology in Switzerland, since it will promote the collaboration between groups with complementary expertise. Through this interaction, a new generation of young investigators will be trained in these areas of research.
Direct link to Lay Summary Last update: 21.02.2013

Responsible applicant and co-applicants

Employees

Publications

Publication
A miR-34a-SIRT6 axis in the squamous cell differentiation network
Lefort Karine, Brooks Yang, Ostano Paola, Cario-Andre Muriel, Calpini Valerie, Guinea-Viniegra Juan, Albinger-Hegyi Andrea, Hoetzenecker Wolfram, Kolfschoten Ingrid, Wagner Erwin F., Werner Sabine, Dotto Gian Paolo (2013), A miR-34a-SIRT6 axis in the squamous cell differentiation network, in EMBO JOURNAL, 32(16), 2248-2263.
Miravirsen (SPC3649) can inhibit the biogenesis of miR-122
Gebert L.F., Rebhan M.A., Crivelli S.E, Denzler R, Stoffel M, Hall J (2013), Miravirsen (SPC3649) can inhibit the biogenesis of miR-122, in Nucleic Acids Res, 1-13.
Harnessing a Physiologic Mechanism for siRNA Delivery With Mimetic Lipoprotein Particles
Nakayama T, Butler JS, Sehgal A, Severgnini M, Racie T, Sharman J, Ding F, Morskaya SS, Brodsky J, Tchangov L, Kosovrasti V, Meys M, Nechev L, Wang G, Peng CG, Fang YP, Maier M, Rajeev KG, Li R, Hettinger J, Barros S, Clausen V, Zhang XM, Wang QF, Hutabarat R (2012), Harnessing a Physiologic Mechanism for siRNA Delivery With Mimetic Lipoprotein Particles, in MOLECULAR THERAPY, 20(8), 1582-1589.
Harnessing a physiologic mechanism for siRNA delivery with mimetic lipoprotein particles
Nakayama T, Butler JS, Sehgal A, Severgnini M, Racie T, Sharman J, Ding F, Morskaya SS, Brodsky J, Tchangov L., Kosovrasti V, Meys M, Nechev L, Wang G, Peng CG, Fan Y, Maier M, Rajeev KG, Li R, Hettinger J, Barros S, Clausen V, Zhang X, Stoffel M, Sah DW (2012), Harnessing a physiologic mechanism for siRNA delivery with mimetic lipoprotein particles, in Molecular Therapy, 20(8), 1582-1589.
Multifocal Epithelial Tumors and Field Cancerization from Loss of Mesenchymal CSL Signaling
Hu B, Castillo E, Harewood L, Ostano P, Reymond A, Dummer R, Raffoul W, Hoetzenecker W, Hofbauer GFL, Dotto GP (2012), Multifocal Epithelial Tumors and Field Cancerization from Loss of Mesenchymal CSL Signaling, in CELL, 149(6), 1207-1220.
IRF6 is a mediator of Notch pro-differentiation and tumour suppressive function in keratinocytes
Restivo G. Nguyen B. C. Dziunycz P. Ristorcelli E. Ryan R. J. Ozuysal O. Y. Di Piazza M. Radt (2011), IRF6 is a mediator of Notch pro-differentiation and tumour suppressive function in keratinocytes, in The EMBO journal, 30(22), 4571-4585.
Nrf2 establishes a glutathione-mediated gradient of UVB cytoprotection in the epidermis
Schafer M. Dutsch S. auf dem Keller U. Navid F. Schwarz A. Johnson D. A. Johnson J. A. Werner (2010), Nrf2 establishes a glutathione-mediated gradient of UVB cytoprotection in the epidermis, in Genes & development, 24(10), 1045-1058.
Opposing roles for calcineurin and ATF3 in squamous skin cancer
Wu X. Nguyen B. C. Dziunycz P. Chang S. Brooks Y. Lefort K. Hofbauer G. F. Dotto G. P. (2010), Opposing roles for calcineurin and ATF3 in squamous skin cancer, in Nature, 465(7296), 368-372.
A novel function of the Nrf2 transcription factor in cornifiedenvelope formation and skin barrier function
Schäfer M. Farwanah H. Willrodt A.-H. Hübner A. Roop.D. Sandhoff K. Bloch W. and Wer, A novel function of the Nrf2 transcription factor in cornifiedenvelope formation and skin barrier function, in submitted, 2011, not yet in press.
Identification of UV-protective Activators of Nuclear Factor Erythroid-derived 2-Related Factor 2 (Nrf2) by Combining a Chemical Library Screen with Computer-based Virtual Screening.
Lieder Franziska, Reisen Felix, Geppert Tim, Sollberger Gabriel, Beer Hans-Dietmar, auf dem Keller Ulrich, Schäfer Matthias, Detmar Michael, Schneider Gisbert, Werner Sabine, Identification of UV-protective Activators of Nuclear Factor Erythroid-derived 2-Related Factor 2 (Nrf2) by Combining a Chemical Library Screen with Computer-based Virtual Screening., in J Biol Chem, 287(39), 33001-33013.
Src is activated by the nuclear receptor peroxisome proliferator-activated receptor β/δ in ultraviolet radiation-induced skin cancer
Montagner A, Delgado MB, Tallichet-Blanc C, Chan JS, Sng MK, Mottaz H, Degueurce G, Lippi Y, Moret C, Baruchet M, Antsiferova M, Werner S, Hohl D, Saati TA, Farmer PJ, Tan NS, Michalik L, Wahli W, Src is activated by the nuclear receptor peroxisome proliferator-activated receptor β/δ in ultraviolet radiation-induced skin cancer, in EMBO Mol Med, 5, 1-19.

Collaboration

Group / person Country
Types of collaboration
CNIO Spain (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Université de Bordeaux France (Europe)
- in-depth/constructive exchanges on approaches, methods or results

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
2nd Faculty and Staff Retreat of the Lausanne Cancer Research Community Individual talk 29.10.2013 Ecole Hôtelière, Lausanne, Switzerland Lefort Karine; Dotto Gian-Paolo;
Gordon Conference Talk given at a conference 15.06.2013 New London, United States of America Werner Sabine;
IID, International Investigative Dermatology 2013 Poster 08.05.2013 Edinburgh, Great Britain and Northern Ireland Lefort Karine;
CSHL meetings and courses Poster 30.10.2012 Cold Spring Harbor, United States of America D'Errico Ilenia;


Self-organised

Title Date Place
Sinergia follow-up meetings (May 2009/ Nov. 2010 (Lausanne); May 2011 (Zürich); Dec. 2011 and July 2012 (Bern) 01.05.2013 Lausanne, Zürich, Bern, Switzerland
Summer course in Oncology 03.09.2012 Epalinges, DB-UNIL, Switzerland
Sinergia Journal Clubs (February, March, May, June 2012) 02.02.2012 Skype, Switzerland

Awards

Title Year
Poster Prize, EMBO workshop on «The reciprocal interactions of signaling pathways and non-coding RNA 2012

Associated projects

Number Title Start Funding scheme
150837 MicroSPECT/PET/CT for preclinical molecular imaging 01.12.2013 R'EQUIP
138653 Control of Notch1 Gene Transcription in skin homeostasis and carcinogenesis 01.10.2011 Project funding (Div. I-III)

Abstract

The skin is an organ of great clinical relevance. It also provides an excellent model system to understand integrated control of tissue homeostasis under normal conditions, in response to external insults (most notably UV light), in the wound healing process, during aging, and in tumorigenesis. It is becoming increasingly clear that complex biological systems such as the skin are intrinsically “robust”, i.e. organized as scale-free networks, with most connections being made to critical “hubs” (or signaling centers). Several “hubs” that integrate and orchestrate the response of the skin to UV irradiation have been identified, and they include transcriptional regulators like p53 and its target Notch1, Nrf2 and PPARß/d. In concert with protein-encoding genes, microRNAs (miRNAs) play a key role in integrating multiple signaling inputs and coordinating the overall gene expression response of cells and tissues to changing environmental conditions. The main goal of this proposal is to explore the role that miRNAs play as integrating determinants of the UVB response of keratinocytes. More specifically, we will test the hypothesis that miRNAs control the acute UVB response of keratinocytes, impinging on intrinsic regulation of these cells and their modulation of other cell types, in a possible cross-talk with the p53/Notch, Nrf2 and PPARß/d pathways. We will further test the hypothesis that miRNAs play an essential function in the long-term UVB response of keratinocytes, in particular in skin cancer development.Using microarray analysis we have identified UVB-regulated miRNAs in acutely and chronically UVB-irradiated mouse skin as well as in human primary keratinocytes. Their roles in UV-mediated cell cycle arrest, differentiation, apoptosis, ROS production, and inflammation will be analyzed, as well as their capability to influence keratinocyte tumor formation. This will be achieved by transfection of keratinocytes with miRNA precursors (oligomiRs) and miRNA blocking oligonucleotides (antagomiRs) in conventional and organotypic culture conditions, human skin explants, and in vivo in mice. For the latter, novel strategies for in vivo delivery of miRNAs/antagomiRs to the skin will be developed. Putative targets of relevant miRNAs will be identified by combined bioinformatic and biochemical approaches, followed by in vitro and in vivo validation. Finally, cross-regulation between specific miRNAs and the p53/Notch1, PPARß/d, and Nrf2 pathways will be determined.These ambitious goals shall be achieved by a close collaboration between three groups with strong and complementary expertise in skin biology, and a fourth group with strong expertise in miRNA biology/technology. Additional expertise in bioinformatics is available through collaboration with an outside expert. The project is expected to shed light onto the mechanisms involved in the UV response of the skin and to identify key regulators involved in skin protection/repair after UV-induced damage. By bringing our laboratories together, we can use a combined comparative approach of the mouse and human systems, which is likely to produce significant novel insight with strong implications for human health. In addition, this project will strengthen the fields of skin biology/ miRNA biology in Switzerland, since it will promote the collaboration between groups with complementary expertise. Through this interaction, a new generation of young investigators will be trained in these areas of research.
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