Impact of preanalytical conditions on functional cancer biomarkers

PROJECT SUMMARYABSTRACT An attractive approach to improve the survival of ovarian cancer patients is to identify the ideal therapy for each tumor. Functional precision medicine (FPM) represents an emerging class of biomarker where the ideal therapy is selected by directly exposing patient tumor cells to potential drug options followed by molecular or phenotypic measurement of drug response. Ideally, FPM biomarkers require viable patient cancer cells that are also phenotypic surrogates of the patient tumor. However, the impact that different pre-analytic tumor handling protocols have on the fidelity and output of FPM biomarkers remains poorly understood. Here, we propose to identify optimal pre-analytic tumor handling conditions for a FPM biomarker. The FPM biomarker used in this proposal is ramp up dynamic BH3 profiling (RUDBP) which measures whether a rapid (2- 16 hour) exposure of chemical libraries induces apoptotic signaling in cancer cells. Importantly RUDBP can be performed on samples with low cell numbers to evaluate drug sensitivity such as core needle biopsies. In this proposal, using human surgical resections and patient derived xenografts, we will measure how pre-analytic tumor handling alters RUDBP. Specific pre-analytic handling conditions include live tumor transport conditions (Aim 1), long-term frozen storage conditions (Aim 2), and pre-analytic ex vivo expansion culture conditions (Aim 3). In each aim we also seek to correct pre-analytic artifacts using mathematical models to align RUDBP of the handled tumor to RUDBP of the fresh tumor. The proposed studies will systematically establish optimal tumor handling conditions for RUDBP by identifying ideal live tumor transport conditions, frozen storage conditions and pre-analytic ex vivo cell culture conditions. We anticipate that careful analysis of pre-analytic conditions will minimize potential false results of functional biomarkers. Finally, we expect that these results will accelerate incorporation of FPM into the clinic, increase accessibility for patients at distant sites, and ultimately aid in identifying ideal therapies for relapsed and refractory ovarian cancer patients.