Project 8

Urokinase pathway and ExtraCellular Matrix remodelling in experimental models of inflammation

ESR8: Saatvik Potluri

Centre National de la Recherche Scientifique – CNRS, Laboratoire Matière et Systèmes Complexes (France)

Supervisors: Dr N. Peyriearas, co-supervisors: local: Dr M. Frain, external: academic: G. Lutfalla (P1, UM), non academic: F. Schmitt (Zeiss)

Objectives: Investigate RNA and protein expression patterns in toto with single cell resolution for urokinase uPA, its inhibitors PAIs and its receptor uPAR (soluble & mb bound) in response to inflammatory episodes evoked by different triggers (infectious and non-infectious) in wild type and perturbed conditions. The activity of uPA will be tuned spatially and temporally with the photoactivation of its caged inhibitor B428. Modulate locally the genetic expression of the Urokinase pathway components with optogenetic tools (cyclofen-OH uncaging). Correlate these data with extracellular matrix remodelling and activated neutrophil and macrophage behaviours. To this end, we shall exploit high resolution in vivo imaging over protracted time periods to accurately map the inflammatory process using fluorescent staining with specific nanobodies and transgenic reporters. Because all metazoans rely on ECM or their functional homologs for the integrity of tissues and on ECM remodeling to allow cells to migrate and tissues to form, we may consider other experimental models of more primitive metazoans to complement our zebrafish work.

Expected Results: 1) Spatiotemporal expression patterns (quantitative multiplex fluorescent RNA in situ hybridization and immnucytochemistry with specific nanobodies) of Urokinase and the component of its pathway during inflammatory episodes (infectious and non-infectious). 2) Highlight their contribution to extracellular matrix remodelling and cell behaviours in wild type and mutant backgrounds (impaired expression of uPA pathway components, optogenetics or inhibitors). 3)Drug interfering (screens) 4) Highlight the evolutionary origin.

Secondments: Academic: P1 Montpellier: Dr L. Yatime, Month 20, 1 month, 3D structures; P4 STUBA: Z. Kriva, Month 24, 1 month, image analysis; P13 RUMC: P. Friedl, Month 30, 1 month, multimodal imaging. Industrial: P12 Acquifer: Dr. J. Gehrig, Month 24, 1 month, screens; P14 LaVision BioTec: Dr. J. Heidelin, Month 18, 1 month, new microscope setup.