Development of a Single-cell Label-free Drug Testing Platform using Fluorescence Lifetime Imaging Microscopy (Flim) for Patients with Metastatic Cancer

Abstract

In the United States, of cancers that affect both men and women, colorectal cancer (CRC) is the second leading cause of cancer-related deaths, resulting in over 50,000 deaths each year. Despite major advances in treatment (i.e., surgery, radiation therapy, chemotherapy, biologic/immune therapy) and resultant improved outcomes over time, it has been demonstrated that failure of treatment in patients with metastatic breast cancer portends a poor prognosis. As the number of potential FDA-approved cancer drugs increases, the likelihood of identifying effective combination strategies tailored to an individual patient's tumor increases. Therefore, to provide significant advances in cancer treatment and enable the most effective chemotherapy, development of more quantitative and objective means for assessing drug sensitivity of an individual patient's tumor that is safer, faster, sensitive and capable for single- or multi-drug testing is urgently needed for advancing personalized therapies. To develop such an approach, fluorescence spectroscopic technologies may serve as a non- invasive means of revealing cell health. We applied phasor-fluorescence lifetime imaging microscopy (phasor- FLIM) to measure drug response using human colorectal cancer cell line HCT116 and patient samples. FLIM can be served as a promising new approach for reading the metabolic states of cancer cells treated with anti-cancer therapeutics and identified specific alterations of metabolic states that are early indicators of irreversible cell demise and that such metabolic changes can be detected using a label-free imaging assay for single-cell drug testing of rare and/or heterogeneous cancer cells. Based on these findings, we apply the label-free imaging platform to measure drug response using patient samples.