Project Grant R43GM146508

HIGH THROUGHPUT SINGLE CELL LINEAR DISPLACEMENT ADHESION ASSAY - ABSTRACT CELL ADHESION IS AN ESSENTIAL PROCESS FOR ANY LIVING CELL. IT IS CRITICAL FOR CELL DIFFERENTIATION, DIVISION, MIGRATION AND SPECIALIZATION. DYSFUNCTION OF CELL ADHESION IS A HALLMARK OF VARIOUS PATHOLOGICAL PHENOTYPES, INCLUDING CHRONIC KIDNEY DISEASES, CANCER, AND MANY OTHERS. IN THE EFFORT TO DISCOVER NEW DISEASE TREATMENTS AND IMPROVE OUR BASIC UNDERSTANDING OF SINGLE CELL PROPERTIES THERE IS A STRONG DEMAND FOR METHODS PERMITTING RAPID AND ACCURATE SINGLE CELL ADHESION MEASUREMENTS. UNFORTUNATELY, CURRENT TECHNOLOGIES ARE LACKING SINCE THEY ONLY MEASURE ADHESION IN ENTIRE CELL POPULATIONS (E.G., MICROFLUIDICS, SPIN DISKS) RATHER THAN INDIVIDUAL CELLS, ARE TOO COMPLEX OR COSTLY (E.G., ATOMIC FORCE MICROSCOPY, OPTICAL TWEEZERS) FOR WIDESPREAD ADOPTION, REQUIRE SOPHISTICATED FUNCTIONALIZED SURFACES (TRACTION FORCE MICROSCOPY), OR PROBE ONLY TARGETED RECEPTORS (CELL ADHESIVE FORCE MICROSCOPY) THAT PRECLUDES MEASURING TOTAL CELL ADHESION POTENTIAL. THEREFORE, THERE IS A GREAT NEED FOR AN AUTOMATED, HIGH-THROUGHPUT AND COST-EFFICIENT PLATFORM CAPABLE OF SIMULTANEOUSLY MEASURING SINGLE CELL ADHESION FOR A LARGE POPULATION OF CELLS. HERE, WE PROPOSE TO DEVELOP A NOVEL METHODOLOGICAL APPROACH UTILIZING SIMPLE DISPOSABLE MICROFLUIDICS CASSETTES (MICS), OSCILLATION DRIVEN CELL SHIFTS, AND A CONVERSION OF EACH INDIVIDUAL CELL TRACK TO ADHESION FORCE VIA MACHINE LEARNING ALGORITHMS. OUR INITIAL STUDIES PROVIDE STRONG SUPPORT FOR THE FEASIBILITY OF THE APPROACH. THIS ONE-YEAR PHASE I PROJECT WILL RESULT IN A FULLY FUNCTIONAL INSTRUMENT THROUGH THE DEVELOPMENT AND INTEGRATION OF ITS CRITICAL COMPONENTS, WHICH INCLUDE A PROGRAMMABLE CELL SHIFT ACTUATOR (BIOSHAKE), DISPOSABLE MICS, CELL ADHESION ANALYSIS ALGORITHMS, AND SOFTWARE (SA1). IN ADDITION, THE ENTIRE WORKFLOW AND PROOF OF PRINCIPLE EXPERIMENTS WILL BE PERFORMED USING MULTIPLE CELL LINES WITH VARIOUS ADHESIVE PROPERTIES (SA2). WE ANTICIPATE THAT THE FULLY DEVELOPED MATURE PRODUCT WILL PROVIDE A HIGH IMPACT TOOL TO PROMOTE MECHANOBIOLOGY STUDIES ON THE KEY ROLE OF CELL ADHESION IN HEALTH AND DISEASE, INCLUDING SUCH PATHOLOGICAL CONDITIONS, SUCH AS CANCER, THUS FACILITATING FURTHER FUNDAMENTAL STUDIES IN CELL BIOLOGY AND TRANSLATIONAL RESEARCH.