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Acoustic enrichment of heterogenous circulating tumor cells and clusters from patients with metastatic prostate cancer

Magnusson, Cecilia LU ; Augustsson, Per LU ; Undvall Anand, Eva LU ; Lenshof, Andreas LU ; Josefsson, Andreas ; Welén, Karin ; Bjartell, Anders LU ; Ceder, Yvonne LU orcid ; Lilja, Hans LU orcid and Laurell, Thomas LU (2023)
Abstract

BACKGROUND: There are important unmet clinical needs to develop cell enrichment technologies to enable unbiased label-free isolation of both single cell and clusters of circulating tumor cells (CTCs) manifesting heterogeneous lineage specificity. Here, we report a pilot study based on microfluidic acoustophoresis enrichment of CTCs using the CellSearch CTC assay as a reference modality.

METHODS: Acoustophoresis uses an ultrasonic standing wave field to separate cells based on biomechanical properties (size, density, and compressibility) resulting in inherently label-free and epitope-independent cell enrichment. Following red blood cell lysis and paraformaldehyde fixation, 6 mL of whole blood from 12 patients with metastatic... (More)

BACKGROUND: There are important unmet clinical needs to develop cell enrichment technologies to enable unbiased label-free isolation of both single cell and clusters of circulating tumor cells (CTCs) manifesting heterogeneous lineage specificity. Here, we report a pilot study based on microfluidic acoustophoresis enrichment of CTCs using the CellSearch CTC assay as a reference modality.

METHODS: Acoustophoresis uses an ultrasonic standing wave field to separate cells based on biomechanical properties (size, density, and compressibility) resulting in inherently label-free and epitope-independent cell enrichment. Following red blood cell lysis and paraformaldehyde fixation, 6 mL of whole blood from 12 patients with metastatic prostate cancer and 20 healthy controls were processed with acoustophoresis and subsequent image cytometry.

RESULTS: Acoustophoresis enabled enrichment and characterization of phenotypic CTCs (EpCAM
+ , Cytokeratin
+ , DAPI
+ , CD45
- /CD66b
- ) in all patients with metastatic prostate cancer and detected CTC-clusters composed of only CTCs or heterogenous aggregates of CTCs clustered with various types of white blood cells in 9 out of 12 patients. By contrast, CellSearch did not detect any CTC-clusters, but detected comparable numbers of phenotypic CTCs as acoustophoresis, with trends of finding higher number of CTCs using acoustophoresis.

CONCLUSION: Our preliminary data indicate that acoustophoresis provides excellent possibilities to detect and characterize CTC-clusters as a putative marker of metastatic disease and outcomes. Moreover, acoustophoresis enables sensitive label-free enrichment of cells with epithelial phenotype in blood and offers opportunities to detect and characterize CTCs undergoing epithelial-to-mesenchymal transitioning and lineage plasticity.

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author
; ; ; ; ; ; ; ; and
organization
publishing date
type
Working paper/Preprint
publication status
published
subject
publisher
medRxiv
external identifiers
  • pmid:38106097
DOI
10.1101/2023.12.04.23299128
language
English
LU publication?
yes
id
5cb2ebce-d242-4fe0-849e-d6036b79e6ee
date added to LUP
2023-12-27 10:23:31
date last changed
2023-12-27 11:34:15
@misc{5cb2ebce-d242-4fe0-849e-d6036b79e6ee,
  abstract     = {{<p>BACKGROUND: There are important unmet clinical needs to develop cell enrichment technologies to enable unbiased label-free isolation of both single cell and clusters of circulating tumor cells (CTCs) manifesting heterogeneous lineage specificity. Here, we report a pilot study based on microfluidic acoustophoresis enrichment of CTCs using the CellSearch CTC assay as a reference modality.</p><p>METHODS: Acoustophoresis uses an ultrasonic standing wave field to separate cells based on biomechanical properties (size, density, and compressibility) resulting in inherently label-free and epitope-independent cell enrichment. Following red blood cell lysis and paraformaldehyde fixation, 6 mL of whole blood from 12 patients with metastatic prostate cancer and 20 healthy controls were processed with acoustophoresis and subsequent image cytometry.</p><p>RESULTS: Acoustophoresis enabled enrichment and characterization of phenotypic CTCs (EpCAM <br>
 + , Cytokeratin <br>
 + , DAPI <br>
 + , CD45 <br>
 - /CD66b <br>
 - ) in all patients with metastatic prostate cancer and detected CTC-clusters composed of only CTCs or heterogenous aggregates of CTCs clustered with various types of white blood cells in 9 out of 12 patients. By contrast, CellSearch did not detect any CTC-clusters, but detected comparable numbers of phenotypic CTCs as acoustophoresis, with trends of finding higher number of CTCs using acoustophoresis.<br>
 </p><p>CONCLUSION: Our preliminary data indicate that acoustophoresis provides excellent possibilities to detect and characterize CTC-clusters as a putative marker of metastatic disease and outcomes. Moreover, acoustophoresis enables sensitive label-free enrichment of cells with epithelial phenotype in blood and offers opportunities to detect and characterize CTCs undergoing epithelial-to-mesenchymal transitioning and lineage plasticity.</p>}},
  author       = {{Magnusson, Cecilia and Augustsson, Per and Undvall Anand, Eva and Lenshof, Andreas and Josefsson, Andreas and Welén, Karin and Bjartell, Anders and Ceder, Yvonne and Lilja, Hans and Laurell, Thomas}},
  language     = {{eng}},
  month        = {{12}},
  note         = {{Preprint}},
  publisher    = {{medRxiv}},
  title        = {{Acoustic enrichment of heterogenous circulating tumor cells and clusters from patients with metastatic prostate cancer}},
  url          = {{http://dx.doi.org/10.1101/2023.12.04.23299128}},
  doi          = {{10.1101/2023.12.04.23299128}},
  year         = {{2023}},
}