At present, the administration of chemotherapy drugs is largely based on physician experience and clinical guidelines. Clearly, a personalized solution would significantly benefit patients in the improvement of clinical outcomes. In this work, we used a spectroscopic approach in which a water-soluble tetrazolium salt (WST-8) assay was applied to patient-derived slice organoids. WST-8 would react with intracellular dehydrogenase resulting in the formation of formazan dye. The absorption at 450nm can be used to assess the viability of the organoids 5 days after initiation of organoid culture. With additional development, this approach may be used to improve the clinical outcome of patients undergoing chemotherapy.
In this study, we performed two-photon imaging of porcine skin and oral mucosa specimens under simulated tissue glycation. After 14 days of glycation treatment with fructose, AGEs were formed in both porcine skin and oral mucosa. The autofluorescence response of both skin and mucosa depended on the detection bandwidth with the wavelength region of 330-480 nm demonstrated the strongest detection of AGE autofluorescence.
This work illustrates how tumor proportional score is estimated using object detection method YOLO and compared with a pathologist's calculation. Results show deep learning can achieve good results and be used on clinical applications.
At the present, identifying head and neck squamous cell carcinoma (HNSCC) patients for immune checkpoint inhibitor therapy (ICIT) is achieved through the determination of Tumor Proportion Score (TPS) or the percentage of tumor cells positively labeled for PD-L1. Estimation of TPS is largely done in a manual fashion by a trained pathologist. In the case of HNSCC, the histological section can be over 1 cm in size in which over 100,000 cancer cells need to be evaluated for PD-L1 expression. To expedite the TPS evaluation process for such large specimens, we have developed a platform in which artificial intelligence (AI) is used for TPS determination. With additional development, this approach may be used in the clinical setting to assist pathologists in TPS evaluation.
Since tumor doubling time of head and neck squamous cell carcinoma (HNSCC) is on the order of 100 days, prescribing the most effective treatment for HNSCC patients is of great benefit to the patients. In this work, we describe the use of precisely cut tumor slices on the order of 200 mm in thickness for testing the effect of chemotherapy drugs. Both spectroscopic and microscopic imaging were used to analyze treatment efficacy. Overall, viability of tumor slices decreases with increased drug concentration although within the same patient, drug response varies among different tumor slices. With additional development, this approach may be used to predict treatment of chemotherapy and immunotherapy response for personalized medicine.
Traditionally, testing of therapeutic agents uses two-dimensional cell cultures which does not recapitulate the complex, three-dimensional architecture of tissues. In the case of cancer drug development, immunotherapy (IT) emerged as an effective treatment strategy for the patients who respond to therapy. However, the overall patient response rate for immunotherapy is around 25%. Therefore, there is a need to develop more effective drug testing platform for cancer patients. In this work, we develop three-dimensional tissue cultures of human oral cancer slices. We observed the growth dynamics of the tissue sections for up to 28 days. Their viability over extended periods suggest that the tissue sections may be used for personalized drug testing.
Bioimaging of fluorescent probes provides a fantastic tool to visualize different cellular responses from various cells and to monitor target/probe interaction in cells. Previously, we introduced a small molecule named 3,6-bis(1-methyl-4- vinylpyridinium)carbazole diiodide (BMVC) as a fluorescent marker to light up live cancer cells for cancer diagnosis. Here we used time-gated fluorescence lifetime imaging microscopy (FLIM) of a BMVC isomer, 3,6-bis(1-methyl-2- vinylpyridinium)carbazole diiodide (o-BMVC) as a fluorescent probe to distinguish fixed cancer cells for cancer diagnosis. The time-gated FLIM results showed many more number of o-BMVC foci, characterized by the longer fluorescent decay time of o-BMVC (≥2.4 ns), in fixed cancer cells (HeLa, H1299, CL1-0, MCF-7, MCF-7/ADR, and SAS) than in fixed normal cells (MRC-5, IMR-90, and BJ). Further study of tissue biopsy, a total of 50 head and neck cancer (HNC) samples obtained during surgery and 20 normal oral samples collected from healthy volunteers showed that o-BMVC foci are hardly detectable in the normal oral epithelial cells. The average numbers of o-BMVC foci in tumors and in normal oral epithelial cells are 28.3 and 2.2, respectively. The receiver operating characteristic curve analysis showed the area under curve was 0.992, indicating that this method provides a very high accuracy for clinical detection of HNC cancers. We propose that the combination of BMVC test for live cells and o-BMVC test for fixed cells could provide a powerful tool for screening of human cancers.
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