Fluorescence tomography plugin unit for spatial monitoring of T cell migration

ImmunotherapyIMTis a cancer treatment that harnesses activated T cells to induce a targeted immune response against the cancerPreclinical IMT research has been limited because there are no effective methods to longitudinally image T cell biodistribution in mouse modelsincluding ovarian cancerThis is an urgent unmet need because the only viable methods to monitor activated T cellsthe immune marker most associated with antitumor response and prognosisimmunohistochemistry and FACsare both terminal and ex vivoFluorescence imagingFLioffers many advantages for monitoring T cell migrationincluding the relatively long photostability of fluorescent ligandsease of useand its low costHoweverFLi does not provideD spatial maps of fluorescent reporters due to diffuse light propagation in animal tissueIn additionfluorescence intensities on the tissue surface also depend on the animal s sizeposeand shape andhencelimit quantification and reproducibilityLastthere is no anatomical reference that also provides a template for automated organ delineation along with T cell biodistribution analysisThereforeInVivo Analytics seeks funding to develop InVivoFLUORan automated data analysis tool forD fluorescence tomographyFLtof mouse modelsInVivoFLUOR is comprised ofa Body Conforming Animal MoldBCAMfor multi source transillumination FLt and spatial registration of the animal s geometry and posean Organ Probability MapOPMfor providing an organ templateand a cloud based FLt algorithmWe will demonstrate its feasibility on an IMT example for determining the spatial biodistribution of fluorescence labeled T cells andin combination with bioluminescence imagingBLiwill compare localization of ovarian cancer cellsIn Aimwe will confirm the ability to quantitatively determine fluorescent targets inside a small animalThe spatial distribution of known fluorescent targets will be reconstructed and compared to ex vivo dataIn Aimwe will confirm the ability to determine the in vivo T cell biodistribution at tumor sitesThe fluorescence labeled T cell distribution will be calculated and coregistered to the anatomy based on the OPM and to disseminated ovarian tumorsThe ability to instantaneously quantify the T cell distribution in the same animal longitudinallyas opposed to sacrificing a different animal at every time point for T cell counting via FACS or histologyneither of which can identify sites where the activated T cells may behidinghas an impact on the development of and outcome of new IMTs with high accuracyInVivoFLUOR will enable cross platform data comparison and analysiseliminate operator dependent variabilityincrease data reproducibilityand will facilitate the translation of new therapeuticsThe successful completion of the proposed project will help to commercialize InVivoFLUOR and will find immediate application in the pharmaceutical industry for rapid development of novel IMTs InVivo Analytics seeks funding for demonstrating the feasibility for mapping theD spatial biodistribution of fluorescent T cells in a small animal model of ovarian cancerAnalysis of T cell migration currently lacks the ability to obtainD spatial maps of T cells in the same animal in longitudinal studiesThereforeInVivo Analytics will develop aD fluorescence tomography and data analysis toolwhich can quantify T cell migration in the living animalthereby allowing accurate monitoring of novel immunotherapies