A universal mechanobiology tool for drug discovery in lung diseases

Abstract Despite significant effortsno cure exists for the major respiratory diseases such as asthmaemphysema and pulmonary fibrosis due to a lack of proper preclinical methodsDrug development is often carried out in cultured cellsWhile amenable to imaging and manipulationthis approach lacks the native tissue environment and neglects the effects of mechanical forces due to breathing which influence many signaling pathwaysAlternativelyanimal experiments are low throughput and performed using knock out or transgenic mice which is also limited in its ability to targetimage and pharmacologically manipulate individual cellsThe Precision Cut Lung SlicePCLSis a preparation at an intermediate scale that permits visualization of cells and the extracellular matrix in the native lung tissueHowevercurrent methodsdo not deliver periodic stretch that mimics breathingdo not determine parenchymal or airway wall stiffnessor airway contractile forcefunctional manifestations of lung diseaseandare not suitable for high throughput drug screeningWe propose to designbuild and test a universal high throughput mechanobiological toolcalled the AccuStretchwhich can stretch up toPCLS samples with breathing like waveforms for extended periods of timeandgtdayswhile simultaneously allowing measurement of stiffness and airway contractile force as well as cellular imaging and pharmacological manipulationOur specific aims are as followsAimDesignbuild and test a versatile multi well mechanobiology toolthe AccuStretchmonthsWe will attach gels of known stiffness to the wells and verify the stiffness measurements in the AccuStretchUsing computational finite element analysiswe will fully validate the methodAimStretch rat PCLS samples including an airway with breathing like waveforms and determine parenchymal stiffness and airway contractile force in response to methacholine challengemonthsRat PCLS samples will be prepared and stretched in the AccuStretch for up todays to verify tissue viability and to determine stiffness and airway contractile forceFor comparisonparenchymal stiffness and airway contractile force will be independently measured in tissue strips from the PCLS in an organ bath systemIn this Phase I applicationwe will combine the complementary expertise at Mechanobiologix LLC and Boston University to produce a prototype AccuStretch systemTo achieve these goalsscientists at Mechanobiologix LLC will organize the studiesdesign the AccuStretchcarry out computational simulations and create various test membranesSystem building and testing as well experimental procedures using PCLS will be contracted to Boston UniversityIn Phase IIwe plan to move on toward commercialization by prototype refinement and disease modeling using human PCLSThe end result will be a versatile device that will find countless applications in respiratory as well as other laboratories suitable for basic research as well as drug discovery with an estimated total addressable market of $M worldwide NARRATIVE The major respiratory diseasesasthmapulmonary fibrosis and emphysemacontribute significantly to mortality and morbidity worldwideDespite significant efforts into basic science and drug developmentno cure exists for these diseases due in large part to a lack of proper preclinical methodsThis proposal seeks to designbuild and test a novel screening device that is expected to revitalize respiratory basic research and drug discovery