In situ assay for RNA decapping in tumors

In situ assay for RNA decapping in tumors Abstract This project aims to introduce a new histotechnology to assess tumor cell stress and cell death propensitywhich can be useful in evaluating cancer response to therapyThe adaptive response of tumor cells to stress and their survival in hostile environments are signaled by the levels of mRNA decayIncreased decay dumps important mRNAs and marks cells in distressConverselydecay blockage is highly protectiveIt stabilizes active mRNAs in cancer cellssupports metabolic stability and successful adaptation of the tumorThe major RNA destruction pathway ismRNA decayIts defining step is decappingwhich removes the Nmethyl guanosine cap fromRNA endsRNA decapping is the rate determining step and the best indicator of the decay activityAlthough mRNA decay is a key process in tumor survivalat present it cannot be imaged and assessed in tissue sectionsIn the absence of specific histotechnologiesit is currently studied by using bulk biochemical approaches which have limited value in heterogeneous pathology samplesIn this project we will overcome this obstacle and will develop a new and enabling technology for molecular analysis of tumor cellsWe will introduce the first assay for in situ assessment ofmRNA decay pathway via specific labeling of its decapping step in tissue sectionsThe project will create a new molecular histopathology tool and an advantageous product with wide applicability in studies of cancer and drug development where assessments of mRNA decay are essentialThe proposal will reach these Specific AimsTo develop the first methodology for selective detection of RNA decapping in tissue sectionsThe approach will label the key event of this pathway and visualize the characteristic RNA cleavage activity exposingterminal monophosphatesTo test and optimize the new labeling approach in tissue sections by using several models with active RNA decay reaction including glioblastomaTo validate the new assay specificity and ensure its adequate sensitivity and reliable robust detectionNarrative Statement The purpose of this project is to introduce an enabling method to analyze many varieties of cancerThis will produce a new and useful histological assay with broad commercial applications in the development of more efficient therapies and for molecular research in cancer and other pathologies where tissues sections are routinely used