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- Non-Invasive Profiling of Advanced Prostate Cancer via Multi-Parametric Liquid Biopsy and Radiomic Analysis

# Non-Invasive Profiling of Advanced Prostate Cancer via Multi-Parametric Liquid Biopsy and Radiomic Analysis

Morrison G, Buckley J, Ostrow D, Varghese B, Cen SY, Werbin J, et al.

International Journal of Molecular Sciences . 2022;23(5):2571. DOI [10.3390/ijms23052571](https://doi.org/10.3390/ijms23052571).

How to cite

### AMA

Morrison G, Buckley J, Ostrow D, Varghese B, Cen SY, Werbin J, et al. Non-Invasive Profiling of Advanced Prostate Cancer via Multi-Parametric Liquid Biopsy and Radiomic Analysis. Int J Mol Sci . 2022;23(5):2571. doi:10.3390/ijms23052571

### APA

Morrison, G., Buckley, J., Ostrow, D., Varghese, B., Cen, S. Y., Werbin, J., et al. (2022). Non-Invasive Profiling of Advanced Prostate Cancer via Multi-Parametric Liquid Biopsy and Radiomic Analysis. International Journal of Molecular Sciences , 23(5), 2571. https://doi.org/10.3390/ijms23052571

### BibTeX

@article{morrison2022noninvasive,
title = {Non-Invasive Profiling of Advanced Prostate Cancer via Multi-Parametric Liquid Biopsy and Radiomic Analysis},
author = {Morrison, Gareth and Buckley, Jonathan and Ostrow, Dejerianne and Varghese, Bino and Cen, Steven Y. and Werbin, Jeffrey and others},
journal = {International Journal of Molecular Sciences},
volume = {23},
number = {5},
pages = {2571},
year = {2022},
doi = {10.3390/ijms23052571}
}

In advanced prostate cancer, clinicians need ways to read a tumor&rsquo;s changing biology without repeated tissue biopsies, which grow hard to obtain once the disease has spread. A single blood draw holds several clues at once &mdash; whole tumor cells, free-floating tumor DNA, and even the texture of bone metastases on routine CT scans &mdash; yet these signals are usually studied one at a time.

This study drew one blood sample from 22 men with metastatic castration-resistant prostate cancer and read three layers together: circulating tumor cells, matched cell-free DNA, and CT radiomics. The cellular and DNA readouts agreed where they overlapped, and the imaging texture tracked with how many tumor cells were in circulation.

Reading the layers side by side gave a fuller, non-invasive picture than any single measure alone.

[Read publication at International Journal of Molecular Sciences](https://doi.org/10.3390/ijms23052571)

## Key findings

- The AccuCyte–CyteFinder workflow detected circulating tumor cells in 77% of patients and tracked an established reference method closely. Across 22 men a total of 947 CTCs were counted (median 1.5 per 7.5 mL), and among the 18 patients with paired counts the two methods were highly concordant (R&sup2; = 0.88), with the enrichment-free workflow flagging CTCs in 15 of 18 versus 11 of 18.

- Protein staining sorted the captured tumor cells into treatment-resistance phenotypes that rarely co-occurred. Among 284 CTCs from 12 patients, ARv7 was found in 4 patients (33%) and synaptophysin in 5 (42%), yet only one cell carried both markers.

- Single tumor cells were retrieved and sequenced, and their DNA matched cell-free DNA on the alterations that guide prostate cancer care. Individual CTCs were recovered from 15 patients; somatic variants appeared in CTCs in 92% of sequenced patients versus 45% in cell-free DNA, and androgen-receptor copy-number calls stayed highly consistent between single cells and plasma.

## The AccuCyte–CyteFinder workflow in the methods

&ldquo;Patient blood samples were collected in RareCyte BCTs and processed to plasma and slides using the AccuCyte&reg; Sample Preparation System [66,67]. Resulting slides containing monolayers of nucleated cells were air dried and stored at −20 °C prior to staining. Plasma collected as part of the AccuCyte workflow was placed in microcentrifuge tubes followed by centrifugation at 16,000× g for 10 min. The plasma supernatant was transferred to new centrifuge tubes and stored at −80 °C. 4.3. Immunofluorescence Staining Slides were thawed at room temperature for 30 min, then fixed in 10% neutral buffered formalin for 40 min, followed by two washes in Tris-buffered saline (TBS) for 5 min to neutralize the formalin. Slides were stained either using the RarePlex&reg; 0900-LA CTC Panel Kit (RareCyte, Seattle, WA, USA) for CTC enumeration or using the RarePlex&reg; ARv7 CTC Panel Kit (0913-LB) and Developer Kit for synaptophysin (clone 27G12; Leica Biosystems, Wetzlar, Germany) on a Leica BOND RX automated immunohistochemistry stainer. Stained slides were coverslipped using CyteMount&reg; Mounting Medium (RareCyte) and dried at room temperature for at least one hour prior to scanning. 4.4. Imaging and Analysis Slides were scanned at 10x magnification using the CyteFinder&reg; HT Instrument (RareCyte).&rdquo;

&mdash; Morrison et al., International Journal of Molecular Sciences (2022), Materials and Methods 4.2–4.4, &ldquo;AccuCyte Sample Preparation&rdquo;

Disclosure: RareCyte is listed as an author affiliation on the publication cited above.

Disclosure: RareCyte is named in the competing-interests statement of the publication cited above.

## Why it matters for The AccuCyte–CyteFinder workflow users

If you are weighing an enrichment-free route to circulating tumor cells, this study shows how far a single blood draw can be pushed. The AccuCyte–CyteFinder workflow did more than count cells: it isolated every nucleated cell by density, stained them for prostate-relevant markers, imaged whole slides, then retrieved individual tumor cells for sequencing &mdash; all from the same sample. That end-to-end reach is the point for your own work. Because cells are captured by density rather than selected on a surface antigen, a tumor cell that carries little of that antigen is still on the slide when you look. Counting agreed with an established reference method, so the numbers hold up against an accepted standard. Retrieving single cells then let their DNA be compared directly against matched cell-free DNA, and the two agreed on androgen-receptor changes that steer treatment. When you need counts, phenotype, and single-cell genomics from one tube, this paper is evidence the workflow can carry all three without losing the cells along the way.

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