Preanalytical Variables for the Genomic Assessment of the Cellular and Acellular Fractions of the Liquid Biopsy in a Cohort of Breast Cancer Patients

Stephanie N Shishido, Lisa Welter, Mariam Rodriguez-Lee, Anand Kolatkar, Liya Xu, Carmen Ruiz, Anna S Gerdtsson, Sara Restrepo-Vassalli, Anders Carlsson, Joe Larsen, Emily J Greenspan, E Shelley Hwang, Kathryn R Waitman, Jorge Nieva, Kelly Bethel, James Hicks, Peter Kuhn

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskriftPeer review

8 Citeringar (SciVal)

Sammanfattning

Liquid biopsy allows assessment of multiple analytes, providing temporal information with potential for improving understanding of cancer evolution and clinical management of patients. Although liquid biopsies are intensely investigated for prediction and response monitoring, preanalytic variables are of primary concern for clinical implementation, including categories of collection method and sample storage. Herein, an integrated high-density single-cell assay workflow for morphometric and genomic analysis of the liquid biopsy is used to characterize the effects of preanalytical variation and reproducibility of data from a breast cancer cohort. Following prior work quantifying performance of commonly used blood collection tubes, this study completes the analysis of four time points to assay (24, 48, 72, and 96 hours), demonstrating precision up to 48 hours after collection for assay sensitivity, highly reproducible rare cell enumeration, morphometric characterization, and high efficiency and capacity for single-cell genomic analysis. For the cell-free analysis, both freezing and use of fresh plasma produced similar quality and quantity of cell-free DNA for sequencing. The genomic analysis (copy number variation and single-nucleotide variation) described herein is broadly applicable to liquid biopsy platforms capable of isolating cell-free and cell-based DNA. Morphometric parameters and genomic signatures of individual circulating tumor cells were evaluated in relation to patient clinical response, providing preliminary evidence of clinical validity as a potential biomarker aiding clinical diagnostics or monitoring progression.

Originalspråkengelska
Sidor (från-till)319-337
TidskriftJournal of Molecular Diagnostics
Volym22
Utgåva3
Tidigt onlinedatum2020 jan. 22
DOI
StatusPublished - 2020 mars
Externt publiceradJa

Bibliografisk information

Copyright © 2020 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

Ämnesklassifikation (UKÄ)

  • Biomedicinsk laboratorievetenskap/teknologi

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