Single-Cell Phenotypic and Molecular Characterization of Circulating Tumor Cells Isolated from Cryopreserved Peripheral Blood Mononuclear Cells of Patients with Lung Cancer and Sarcoma.
BACKGROUND: The isolation of circulating tumor cells (CTCs) requires rapid processing of the collected blood due to their inherent fragility. The ability to recover CTCs from peripheral blood mononuclear cells (PBMCs) preserved from cancer patients could allow for retrospective analyses or multicent...
| Publicado en: | Clinical Chemistry Vol. 68; no. 5; pp. 691 - 702 |
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| Autores principales: | , , , , , , , , , , , , , , , , |
| Formato: | Journal Article |
| Publicado: |
Oxford University Press / USA
May2022
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=157165617&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 157165617 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 00099147 10CS jtl: Clinical Chemistry issn: 00099147 maglogo: N pubinfo: dt: May2022 vid: 68 iid: 5 pid: 622 pub: Oxford University Press / USA artinfo: ui: 157165617 10.1093/clinchem/hvac019 157165617 ppf: 691 ppct: 11 formats: fmt: @attributes: type: P tig: atl: Single-Cell Phenotypic and Molecular Characterization of Circulating Tumor Cells Isolated from Cryopreserved Peripheral Blood Mononuclear Cells of Patients with Lung Cancer and Sarcoma. aug: au: Vismara, Marta Reduzzi, Carolina Silvestri, Marco Murianni, Fabio Lo Russo, Giuseppe Fortunato, Orazio Motta, Rosita Lanzoni, Davide Giovinazzo, Francesca Miodini, Patrizia Pasquali, Sandro Suatoni, Paola Pastorino, Ugo Roz, Luca Sozzi, Gabriella Cappelletti, Vera Bertolini, Giulia affil: Biomarkers Unit, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy sug: ab: BACKGROUND: The isolation of circulating tumor cells (CTCs) requires rapid processing of the collected blood due to their inherent fragility. The ability to recover CTCs from peripheral blood mononuclear cells (PBMCs) preserved from cancer patients could allow for retrospective analyses or multicenter CTC studies. METHODS: We compared the efficacy of CTC recovery and characterization using cryopreserved PMBCs vs fresh whole blood from patients with non-small cell lung cancer (NSCLC; n=8) and sarcoma (n=6). Two epithelial cellular adhesion molecule (EpCAM)-independent strategies for CTC enrichment, based on ParsortixVR technology or immunomagnetic depletion of blood cells (AutoMACSVR) were tested, followed by DEPArrayTM single-cell isolation. Phenotype and genotype, assessed by copy number alterations analysis, were evaluated at a single-cell level. Detection of target mutations in CTCenriched samples from frozen NSCLC PBMCs was also evaluated by digital PCR (dPCR). RESULTS: The use of cryopreserved PBMCs from cancer patients allowed for the retrospective enumeration of CTCs and their molecular characterization, using both EpCAM-independent strategies that performed equally in capturing CTC. Cells isolated from frozen PBMCs were representative of whole blood-derived CTCs in terms of number, phenotype, and copy number aberration profile/target mutations. Long-term storage (-3 years) did not affect the efficacy of CTC recovery. Detection of target mutations was also feasible by dPCR in CTC-enriched samples derived from stored PBMCs. CONCLUSIONS: Isolating CTCs from longitudinally collected PBMCs using an unbiased selection strategy can offer a wider range of retrospective genomic/phenotypic analyses to guide patients' personalized therapy, paving the way for sample sharing in multicenter studies. pubtype: Academic Journal doctype: Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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