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Pace, J. S.

Publications and source records attributed to Pace, J. S..

3 recordsLinked to original sources

Machine Learning Approach for Enumeration of Circulating Cells with Diffuse in vivo Flow Cytometry

SignificanceDiffuse in vivo flow cytometry (DiFC) is an emerging technique for enumerating rare, fluorescentlylabeled circulating tumor cells (CTCs) in small animals without drawing blood samples. DiFC uses detection of transient fluorescent peaks in time-series data. Previously, we used a simple amplitude threshold-based method for identifying peak candidates, but it ignores potentially useful information in peak shape that could reduce false-positive detections from instrument noise and increase detection efficiency of lower-amplitude peaks. AimTo develop a machine learning (ML)-integrated signal processing approach for improved CTC enumeration using DiFC by distinguishing CTC peaks from artifacts. ApproachWe developed an ML-integrated approach that incorporates a convolutional neural network (CNN) classifier. The CNN was trained to distinguish CTC peaks from artifacts by analyzing peak amplitude and temporal shape characteristics. Performance was validated on in-silico, control, and CTC-bearing mouse datasets. ResultsThe CNN classifier achieved accuracy, precision, sensitivity, and specificity exceeding 98% on test data. Compared with our previously published threshold-based approach, the ML-integrated method increased the number of correctly identified CTCs and their flow direction while reducing false detections across validation datasets. ConclusionsThe ML-integrated approach significantly improves DiFC CTC enumeration, enabling robustness against artifacts in noisy conditions.

bioengineering↗

Considerations for the use of contrast agents with diffuse in vivo flow cytometry to detect circulating cancer cell populations

SignificanceMetastasis is a leading cause of cancer-related deaths. Disseminated circulating tumor cells (CTCs) through the bloodstream seed metastatic tumors at distant sites. Most methods for enumerating CTCs in humans clinically rely on drawing and analyzing small blood samples, but these may yield inaccurate estimates of CTC burden and cannot measure CTC changes over time. Identification and enumeration of CTCs for experimental or clinical purposes largely rely on marker-driven analyses by flow cytometry. AimIn principle, non-invasive fluorescence enumeration of CTCs directly in vivo could provide a more accurate method for enumerating CTCs. However, this will require specific contrast agent for CTCs. The goal of this work is to define characteristics of useful CTC contrast agents and perform preliminary testing of candidate contrast agents used for fluorescence guided surgery (FGS). ApproachWe evaluated a clinical small-molecule folate receptor targeted contrast agent (OTL38, pafolacianine), a fluorogenic pan-cathepsin contrast agent (VGT-309, abenacianine), and a set of custom designed, small-molecule prostate specific membrane antigen (PSMA) targeted contrast agents. We tested these contrast agents using in vitro cell culture models and in in vivo murine models. ResultsAll tested contrast agents showed high uptake and labeling by target cell lines, but also small but significant labeling of non-cancer blood cells. Contrast agents that exhibited rapid clearance from circulation and the fluorogenic approach resulted in significantly reduced non-specific interfering background fluorescence signals. ConclusionsAlthough the fluorescence contrast agents tested have properties useful for labeling of CTCs, as yet none exhibited the required high specificity. This resulted in some labeling of non-cancer blood cells which presented false-positive CTC counts. Improved contrast agent design and multiplexed use of more than one contrast agent may improve this specificity.

bioengineering↗

PuMA: a papillomavirus genome annotation tool

High-throughput sequencing technologies provide unprecedented power to identify novel viruses from a wide variety of (environmental) samples. The field of viral metagenomics has dramatically expanded our understanding of viral diversity. Viral metagenomic approaches imply that many novel viruses will not be described by researchers who are experts on the genomic organization of that virus. There is a need to develop analytical approaches to reconstruct, annotate, and classify viral genomes. We have developed the papillomavirus annotation tool (PuMA) to provide researchers with a convenient and reproducible method to annotate novel papillomaviruses. PuMA provides an accessible method for automated papillomavirus genome annotation. PuMA currently has a 98% accuracy when benchmarked against the 481 reference genomes in the papillomavirus episteme (PaVE). Finally, PuMA was used to annotate 168 newly isolated papillomaviruses, and successfully annotated 1424 viral features. To demonstrate its general applicability, we developed a version of PuMA that can annotate polyomaviruses.\n\nPuMA is available on GitHub (https://github.com/KVD-lab/puma) and through the iMicrobe online environment (https://www.imicrobe.us/#/apps/puma)

genomics↗