TheprocessofEpithelialtoMesenchymalTransition(EMT)isatransdifferentiationeventinwhichepithelialcells switchtheirphenotypetoamesenchymalone,whichfacilitatesindividualcellmigration,cellinvasion,andtissue assembly.Thisdynamicprocessiscriticalduringembryonicdevelopment,and,whencoupledwiththereverse MET process, facilitates correct spatial and temporal development of organs. While EMT is critical for development,itsmisregulationisimplicatedinmanydiseases,includingcardiacfibrosis,cirrhosis,andcancer. The events that initiate EMT are not well understood, but appear to be linked to changes in the intracellular biochemicalsignalingandgeneexpression,solublefactorsection,autocrinesignaling,mechanicalandparacrine signalingbetweenneighboringcells,andmechanicalsignalingbetweentheepithelialsheetandtheunderlying extracellular matrix (ECM). The Conway lab has demonstrated that a drop in force on adherens junctions is necessary for progression of EMT. The Lemmon lab has demonstrated that progression of EMT requires assemblyoftheECMproteinfibronectin(FN)intonewfibrils,whichcontainbindingsitesforseveralpro-EMT growthfactors.OurrecentstudysuggeststhatFNfibrilsdriveEMTbyclusteringthesegrowthfactorsatthecell surface.ThesefindingshaveledustoahypothesisinwhichEMTisinitiatedbydisruptionofforcesonadherens junctions,whichinturnredistributesforcestotheunderlyingmatrix,initiatesFNfibrilassembly,clusteringsoluble signalsthatpromoteEMTatthecellsurface,anddrivingintracellularbiochemicalsignaling.Inthiswork,we will: 1) develop a computational model that predicts junctional forces, matrix forces and matrix assembly, autocrine and paracrine signaling, and epithelial and mesenchymal cell markers, and uses these to predict spatial localization of EMT in an assembling and confluent cell sheet. Our model will start from a recently developedcell-basedmodelofcell-matrixinteractions,usingthepreviouslydevelopedLemmon-Romermodel of traction force prediction, and extend this to predict force redistribution of cell-matrix forces in neighboring epithelialcellsandtheforcesactingonadherensjunctionsineachcell.Wewillcouplethesemechanicalforces withamodelofintracellularsignalingpathways.2)Wewilluseanovelsuiteoftoolstoestablishanexperimental system that can simultaneously measure junctional forces, matrix forces, FN assembly, and EMT markers. Coloniesofepithelialcellswillbegeneratedusingmicrocontactprinting,whichallowsforrepeatablecolonysize andshape.WewillmeasurespatialregulationofEMTmarkerswithinthecolonieswhilealsoquantifyingboth junctional forces (using a FRET-based biosensor) and cell-matrix forces (using microcontact printing and a substratedeflectiontechnique)todeterminerelationshipbetweenforcecomponents,matrixassemblyandEMT signaling.3)WewillpredicttheeffectsofpotentialEMTinhibitorsandactivatorswiththemodel,thenvalidate the predictive capability of the model using the experimental system. Experimental data will be related to computationalsimulationsforbothprocesses.

Public Health Relevance

Epithelial-Mesenchymal Transition (EMT) is a key transdifferentiation event that is required for embryonic development and wound healing and is misregulated in several disease states including fibrotic disease and cancer. The initiating events that drive EMT require changes in the contractile forces acting on cell-cell and cell-matrix junctions and changes in cell markers. Here we will develop both novel computational and experimentalplatformsthatpredictandmeasurethekeymechanicalandbiochemicalsignalingeventsinthis system to understand how the interactions between mechanics and biochemical signaling drives this crucial process.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM122855-03
Application #
9869902
Study Section
Modeling and Analysis of Biological Systems Study Section (MABS)
Program Officer
Resat, Haluk
Project Start
2018-03-01
Project End
2022-02-28
Budget Start
2020-03-01
Budget End
2021-02-28
Support Year
3
Fiscal Year
2020
Total Cost
Indirect Cost
Name
Virginia Commonwealth University
Department
Engineering (All Types)
Type
Biomed Engr/Col Engr/Engr Sta
DUNS #
105300446
City
Richmond
State
VA
Country
United States
Zip Code
23298