Centrosome amplification is a frequent event in circulating tumor cells from subjects with metastatic breast cancer

Abstract

Centrosome amplification (CA) is a common phenomenon in cancer, promotes genomic stability and cancer evolution, and has been reported to promote metastasis. CA promotes a stochastic gain/loss of chromosomes during cell division, known as chromosomal instability (CIN). However, it is unclear whether CA is present in circulating tumor cells (CTCs), the seeds for metastasis. Here, we surveyed CA in CTCs from human subjects with metastatic breast cancer. CTCs were captured by CD45 exclusion and selection of EpCAM‐positive cells using an exclusion‐based sample preparation technology platform known as VERSA (versatile exclusion‐based rare sample analysis). Centriole amplification (centrin foci> 4) is the definitive assay for CA. However, determination of centrin foci is technically challenging and incompatible with automated analysis. To test if the more technically accessible centrosome marker pericentrin could serve as a surrogate for centriole amplification in CTCs, cells were stained with pericentrin and centrin antibodies to evaluate CA. This assay was first validated using breast cancer cell lines and a nontransformed epithelial cell line model of inducible CA, then translated to CTCs. Pericentrin area and pericentrin area x intensity correlate well with centrin foci, validating pericentrin as a surrogate marker of CA. CA is found in CTCs from 75% of subjects, with variability in the percentage and extent of CA in individual circulating cells in a given subject, similar to the variability previously seen in primary tumors and cell lines. In summary, we created, validated, and implemented a novel method to assess CA in CTCs from subjects with metastatic breast cancer. Such an assay will be useful for longitudinal monitoring of CA in cancer patients and in prospective clinical trials for assessing the impact of CA on response to therapy.

Chromosomal instability sensitizes patient breast tumors to multipolar divisions induced by paclitaxel

“…3), centrosome amplification before therapy may impair focusing of paclitaxel-induced multipolar spindles and provide an additional predictor of response. Our recently developed method to quantitate centrosome amplification in circulating tumor cells from metastatic breast cancer patients ( 46 ) may be useful in this regard. Second, FISH is limited by sampling errors, sectioning artifacts, and analysis of only a subset of chromosomes.…”

Epigenetic alterations impede epithelial-mesenchymal transition by modulating centrosome amplification and Myc/RAS axis in triple negative breast cancer cells

“…The cell lines CAL-51 and MDA-MB-231 have been reported to serve as low and high chromosome instability (CIN) models respectively. However, MDA-MB-231 cells show a significantly higher degree of centrosome amplification 30 . MDA-MB-231 cell line show invasive in vitro, but when introduced directly into the circulation, the cell line has proven useful in experimental metastasis models.…”

High prevalence and dependence of centrosome clustering in mesenchymal tumors and leukemia

“…In fact, combination therapy with chemotherapeutic drugs in triple negative breast cancer (TNBC) have already entered in phase 1 clinical trials which set the potential of the MPS1 inhibition (NCT03328494; NCT05251714). Another potential advantage of the inhibition of the SAC could be the treatment of metastatic TNBC, since circulating tumor cells have high levels of CA and show good clustering capacity (Singh et al, 2020). It is clear by now that CC is an essential mechanism for chromosome instability, which contributes, between other malignant features such as drug resistance, to a high risk of tumor recurrence (Ganem et al, 2009;Holland & Cleveland, 2009;Krämer et al, 2011;Pannu et al, 2015b;Fan et al, 2021).…”