Identification of epithelial and mesenchymal circulating tumor cells in clonal lineage of an aggressive prostate cancer case

Chai, Shoujie; Ruiz-Velasco, Carmen; Naghdloo, Amin; Pore, Milind; Singh, Mohan; Matsumoto, Nicholas; Kolatkar, Anand; Xu, Liya; Shishido, Stephanie N.; Aparicio, Ana; Zurita, Amado J.; Hicks, James; Kühn, Peter

doi:10.1038/s41698-022-00289-1

Cited by 12 publications

(6 citation statements)

References 48 publications

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“…Blood was collected from a patient diagnosed with de novo metastatic prostate cancer following castration resistance as previously described in [ 101 ]. Blood cells were plated on cell-adhesive (Marienfeld) slides and underwent immunofluorescent stained for a mixture anti-human cytokeratins 1, 4, 5 ,6, 8, 10, 13, 18 and 19, CD45, vimentin, and DAPI as previously described in [ 101 ]. Slides were imaged and analyzed as previously described in [ 101 ].…”

Section: Methodsmentioning

confidence: 99%

Cells in the polyaneuploid cancer cell (PACC) state have increased metastatic potential

Mallin

Kim

Choudhury

et al. 2023

Clin Exp Metastasis

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Although metastasis is the leading cause of cancer deaths, it is quite rare at the cellular level. Only a rare subset of cancer cells (~ 1 in 1.5 billion) can complete the entire metastatic cascade: invasion, intravasation, survival in the circulation, extravasation, and colonization (i.e. are metastasis competent). We propose that cells engaging a Polyaneuploid Cancer Cell (PACC) phenotype are metastasis competent. Cells in the PACC state are enlarged, endocycling (i.e. non-dividing) cells with increased genomic content that form in response to stress. Single-cell tracking using time lapse microscopy reveals that PACC state cells have increased motility. Additionally, cells in the PACC state exhibit increased capacity for environment-sensing and directional migration in chemotactic environments, predicting successful invasion. Magnetic Twisting Cytometry and Atomic Force Microscopy reveal that cells in the PACC state display hyper-elastic properties like increased peripheral deformability and maintained peri-nuclear cortical integrity that predict successful intravasation and extravasation. Furthermore, four orthogonal methods reveal that cells in the PACC state have increased expression of vimentin, a hyper-elastic biomolecule known to modulate biomechanical properties and induce mesenchymal-like motility. Taken together, these data indicate that cells in the PACC state have increased metastatic potential and are worthy of further in vivo analysis.

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Section: Methodsmentioning

confidence: 99%

Cells in the polyaneuploid cancer cell (PACC) state have increased metastatic potential

Mallin

Kim

Choudhury

et al. 2023

Clin Exp Metastasis

View full text Add to dashboard Cite

show abstract

“…Blood cells were plated on cell-adhesive (Marienfeld) slides and underwent immunofluorescent stained for a mixture anti-human cytokeratins 1,4,5,6,8,10,13,18 and 19, CD45, Vimentin, and DAPI as previously described in [88]. Slides were imaged and analyzed as previously described in [88]. b.…”

Section: Ctc Detectionmentioning

confidence: 99%

“…Blood was collected from a patient diagnosed with de novo metastatic prostate cancer following castration resistance as previously described in [88]. Blood cells were plated on cell-adhesive (Marienfeld) slides and underwent immunofluorescent stained for a mixture anti-human cytokeratins 1,4,5,6,8,10,13,18 and 19, CD45, Vimentin, and DAPI as previously described in [88]. Slides were imaged and analyzed as previously described in [88].…”

Section: Ctc Detectionmentioning

confidence: 99%

Cells in the Polyaneuploid Cancer Cell (PACC) state have increased metastatic potential

Mallin

Kim

Choudhury

et al. 2022

Preprint

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Although metastasis is the leading cause of cancer death, it is quite rare at the cellular level. Only a rare subset of cancer cells (~1 in 1.5 billion) can complete the entire metastatic cascade: invasion, intravasation, survival in the circulation, extravasation, and colonization (i.e. are metastasis competent). We propose that cells engaging a Polyaneuploid Cancer Cell (PACC) phenotype are metastasis competent. PACCs are enlarged, non-dividing cells with increased genomic content that form in response to stress. Single-cell tracking using time-lapse microscopy reveals that PACCs are more motile than nonPACCs. Additionally, PACCs exhibit increased capacity for environment-sensing and directional migration in chemotactic environments, predicting successful invasion. Magnetic Twisting Cytometry and Atomic Force Microscopy reveal that cells in the PACC state display hyper-elastic properties like increased peripheral deformability and maintained peri-nuclear cortical integrity that predict successful intravasation and extravasation. Furthermore, four orthogonal methods reveal that PACCs have increased expression of Vimentin, a known hyper-elastic biomolecule. Lastly, anoikis-resistance assays and detection of PACCs in the blood of a patient with metastatic castrate-resistant prostate cancer using a selection-free circulating tumor cell detection platform reveal that PACCs are capable of surviving in the circulation. Taken together with the knowledge that PACCs are capable of eventual depolyploidization and progeny formation (as a potential route to colonization), these data support PACCs as candidate metastasis-competent cells worthy of further analysis.

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“…[46,47] In addition, vimentin is also present at the extracellular surface of endothelial cells and macrophages, [48][49][50] which could potentially appear in the blood and exhibit CD45staining. The definition of mesenchymal CTCs in our study is based on the current literature, which suggests that VIM expression is associated with mesenchymal-like properties in cancer cells, [29,[51][52][53][54][55][56][57][58] including increased motility, invasiveness, and resistance to apoptosis. However, we understand that this definition may not be specific to CTCs, as other rare circulating cells in the blood may also express VIM.…”

Section: Discussionmentioning

confidence: 99%

Automated Image Analysis for Characterization of Circulating Tumor Cells and Clusters Sorted by Magnetic Levitation

Ogut

Gupta

et al. 2023

Advanced Biology

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Magnetic levitation‐based sorting technologies have revolutionized the detection and isolation of rare cells, including circulating tumor cells (CTCs) and circulating tumor cell clusters (CTCCs). Manual counting and quantification of these cells are prone to time‐consuming processes, human error, and inter‐observer variability, particularly challenging when heterogeneous cell types in 3D clusters are present. To overcome these challenges, we developed "Fastcount," an in‐house MATLAB‐based algorithm for precise, automated quantification and phenotypic characterization of CTCs and CTCCs, in both 2D and 3D. Fastcount is 120 times faster than manual counting and produces reliable results with a ±7.3% deviation compared to a trained laboratory technician. By analyzing 400 GB of fluorescence imaging data, we showed that Fastcount outperforms manual counting and commercial software when cells are aggregated in 3D or staining artifacts are present, delivering more accurate results. We further employed Fastcount for automated analysis of 3D image stacks obtained from CTCCs isolated from colorectal adenocarcinoma and renal cell carcinoma blood samples. Interestingly, we observed a highly heterogeneous spatial cellular composition within CTCCs, even among clusters from the same patient. Overall, Fastcount can be employed for various applications with lab‐chip devices, such as CTC detection, CTCC analysis in 3D and cell detection in biosensors.

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Identification of epithelial and mesenchymal circulating tumor cells in clonal lineage of an aggressive prostate cancer case

Cited by 12 publications

References 48 publications

Cells in the polyaneuploid cancer cell (PACC) state have increased metastatic potential

Cells in the polyaneuploid cancer cell (PACC) state have increased metastatic potential

Cells in the Polyaneuploid Cancer Cell (PACC) state have increased metastatic potential

Automated Image Analysis for Characterization of Circulating Tumor Cells and Clusters Sorted by Magnetic Levitation

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