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      Platelet cloaking of circulating tumour cells in patients with metastatic prostate cancer: Results from ExPeCT, a randomised controlled trial

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          Abstract

          Background

          Circulating tumour cells (CTCs) represent a morphologically distinct subset of cancer cells, which aid the metastatic spread. The ExPeCT trial aimed to examine the effectiveness of a structured exercise programme in modulating levels of CTCs and platelet cloaking in patients with metastatic prostate cancer.

          Methods

          Participants (n = 61) were randomised into either standard care (control) or exercise arms. Whole blood was collected for all participants at baseline (T0), three months (T3) and six months (T6), and analysed for the presence of CTCs, CTC clusters and platelet cloaking. CTC data was correlated with clinico-pathological information.

          Results

          Changes in CTC number were observed within group over time, however no significant difference in CTC number was observed between groups over time. Platelet cloaking was identified in 29.5% of participants. A positive correlation between CTC number and white cell count (WCC) was observed (p = 0.0001), in addition to a positive relationship between CTC clusters and PSA levels (p = 0.0393).

          Conclusion

          The presence of platelet cloaking has been observed in this patient population for the first time, in addition to a significant correlation between CTC number and WCC.

          Trial registration

          ClincalTrials.gov identifier NCT02453139.

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          Most cited references34

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          Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases.

          The purpose of this study was to determine the accuracy, precision, and linearity of the CellSearch system and evaluate the number of circulating tumor cells (CTCs) per 7.5 mL of blood in healthy subjects, patients with nonmalignant diseases, and patients with a variety of metastatic carcinomas. The CellSearch system was used to enumerate CTCs in 7.5 mL of blood. Blood samples spiked with cells from tumor cell lines were used to establish analytical accuracy, reproducibility, and linearity. Prevalence of CTCs was determined in blood from 199 patients with nonmalignant diseases, 964 patients with metastatic carcinomas, and 145 healthy donors. Enumeration of spiked tumor cells was linear over the range of 5 to 1,142 cells, with an average recovery of >/=85% at each spike level. Only 1 of the 344 (0.3%) healthy and nonmalignant disease subjects had >/=2 CTCs per 7.5 mL of blood. In 2,183 blood samples from 964 metastatic carcinoma patients, CTCs ranged from 0 to 23,618 CTCs per 7.5 mL (mean, 60 +/- 693 CTCs per 7.5 mL), and 36% (781 of 2,183) of the specimens had >/=2 CTCs. Detection of >/=2 CTCs occurred at the following rates: 57% (107 of 188) of prostate cancers, 37% (489 of 1,316) of breast cancers, 37% (20 of 53) of ovarian cancers, 30% (99 of 333) of colorectal cancers, 20% (34 of 168) of lung cancers, and 26% (32 of 125) of other cancers. The CellSearch system can be standardized across multiple laboratories and may be used to determine the clinical utility of CTCs. CTCs are extremely rare in healthy subjects and patients with nonmalignant diseases but present in various metastatic carcinomas with a wide range of frequencies.
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            Physical activity and survival after prostate cancer diagnosis in the health professionals follow-up study.

            To determine whether higher physical activity after prostate cancer (PCa) diagnosis decreases risk of overall and PCa-specific death. We evaluated physical activity in relation to overall and PCa mortality among 2,705 men in the Health Professionals Follow-Up Study diagnosed with nonmetastatic PCa observed from 1990 to 2008. Proportional hazards models were used to evaluate physical activity and time to overall and PCa-specific death. Among men who lived at least 4 years after their postdiagnosis physical activity assessment, we documented 548 deaths, 20% of which were a result of PCa. In multivariable analysis, men who were physically active had lower risk of all-cause mortality (P(trend) < .001) and PCa mortality (P(trend) = .04). Both nonvigorous activity and vigorous activity were associated with significantly lower overall mortality. Those who walked ≥ 90 minutes per week at a normal to very brisk pace had a 46% lower risk of all-cause mortality (hazard ratio [HR] 0.54; 95% CI, 0.41 to 0.71) compared with shorter durations at an easy walking pace. Men with ≥ 3 hours per week of vigorous activity had a 49% lower risk of all-cause mortality (HR, 0.51; 95% CI, 0.36 to 0.72). For PCa-specific mortality, brisk walking at longer durations was suggestively inverse but not statistically significant. Men with ≥ 3 hours per week of vigorous activity had a 61% lower risk of PCa death (HR, 0.39, 95% CI, 0.18 to 0.84; P = .03) compared with men with less than 1 hour per week of vigorous activity. Men exercising vigorously before and after diagnosis had the lowest risk. In men with PCa, physical activity was associated with lower overall mortality and PCa mortality. A modest amount of vigorous activity such as biking, tennis, jogging, or swimming for ≥ 3 hours a week may substantially improve PCa-specific survival.
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              Circulating tumor cells as a window on metastasis biology in lung cancer.

              Circulating tumor cell (CTC) number in metastatic cancer patients yields prognostic information consistent with enhanced cell migration and invasion via loss of adhesion, a feature of epithelial-to-mesenchymal transition (EMT). Tumor cells also invade via collective migration with maintained cell-cell contacts and consistent with this is the circulating tumor microemboli (CTM; contiguous groups of tumor cells) that are observed in metastatic cancer patients. Using a blood filtration approach, we examined markers of EMT (cytokeratins, E-cadherin, vimentin, neural cadherin) and prevalence of apoptosis in CTCs and CTM to explore likely mechanism(s) of invasion in lung cancer patients and address the hypothesis that cells within CTM have a survival advantage. Intra-patient and inter-patient heterogeneity was observed for EMT markers in CTCs and CTM. Vimentin was only expressed in some CTCs, but in the majority of cells within CTM; E-cadherin expression was lost, cytoplasmic or nuclear, and rarely expressed at the surface of the cells within CTM. A subpopulation of CTCs was apoptotic, but apoptosis was absent within CTM. This pilot study suggests that EMT is not prosecuted homogeneously in tumor cells within the circulation of lung cancer patients and that collective migration and enhanced survival of cells within CTM might contribute to lung cancer metastasis. Multiplex analysis and further detailed exploration of metastatic potential and EMT in CTCs/CTM is now warranted in a larger patient cohort. Copyright © 2011 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.
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                Author and article information

                Contributors
                Role: InvestigationRole: MethodologyRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: ConceptualizationRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Formal analysisRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Formal analysisRole: Writing – review & editing
                Role: ConceptualizationRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: ConceptualizationRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: ConceptualizationRole: Writing – review & editing
                Role: ConceptualizationRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: ConceptualizationRole: InvestigationRole: MethodologyRole: SupervisionRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                18 December 2020
                2020
                : 15
                : 12
                : e0243928
                Affiliations
                [1 ] Department of Histopathology and Morbid Anatomy, Trinity Translational Medicine Institute, Trinity College Dublin, Dublin, Ireland
                [2 ] Department of Histopathology, Cork University Hospital, Cork, Ireland
                [3 ] Department of Pathology, University College Cork, Cork, Ireland
                [4 ] Discipline of Physiotherapy, School of Medicine, Trinity College Dublin, Dublin, Ireland
                [5 ] School of Medicine, Trinity College Dublin, Dublin, Ireland
                [6 ] Pacific Northwest National Laboratory, Richland, Washington, United States of America
                [7 ] Institute of Pathology, University Hospital Basel, Basel, Switzerland
                [8 ] Cancer Trials Ireland, Dublin, Ireland
                [9 ] Centre for Cancer Research and Cell Biology, Queen’s University Belfast, Belfast, Northern Ireland, United Kingdom
                [10 ] King’s College London, School of Cancer and Pharmaceutical Sciences, Translational Oncology and Urology (TOUR), London, United Kingdom
                [11 ] Guy’s and St Thomas’ NHS Foundation Trust, London, United Kingdom
                [12 ] Harvard T.H. Chan school of Public Health, Boston, Massachusetts, United States of America
                [13 ] Department of Radiation Oncology, St Luke’s Hospital, Dublin, Ireland
                [14 ] Department of Oncology, Beaumont Hospital, Dublin, Ireland
                [15 ] Department of Urology, St James’s Hospital, Dublin, Ireland
                [16 ] Department of Surgery, Trinity College Dublin, Dublin, Ireland
                [17 ] Department of Oncology, Mater Misericordiae Hospital, Dublin, Ireland
                [18 ] HOPE Directorate, St James’s Hospital, Dublin, Ireland
                [19 ] Department of Histopathology, St James’s Hospital, Dublin, Ireland
                [20 ] Department of Oncology, Tallaght University Hospital, Dublin, Ireland
                Hospital de Santa Maria, PORTUGAL
                Author notes

                Competing Interests: All authors have completed the ICMJE uniform disclosure form at www.icmje.org/coi_disclosure.pdf and declare: support from the World Cancer Research fund. AMB declares board membership of LUCE which has received support from Abbvie, Amgen, BMS, BI, Lilly, Merck, MSD, Novartis, Pfizer, Roche and Takeda, advisory boards for BMS, Takeda and Pfizer and personal fees and non-financial support from Roche, personal fees and non-financial support from MSD, outside the submitted work. BS reports funding from Trinity College Dublin throughout the conduct of this study. EG and MVH report membership of the editorial board of BMC Cancer. RMD reports personal fees from Clovis, grants and personal fees from Pfizer, grants from Amgen, personal fees and other from Bristol Myers Squibb, other from Merck, Bayer and Janssen and grants from Celgene outside the submitted work. SF reports personal fees and non-financial support from Pfizer, grants, personal fees and non-financial support from MSD, personal fees and non-financial support from Roche, grants, personal fees and non-financial support from Astellas, outside the submitted work. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

                Author information
                https://orcid.org/0000-0002-7876-5164
                https://orcid.org/0000-0003-3437-744X
                Article
                PONE-D-20-21500
                10.1371/journal.pone.0243928
                7748139
                33338056
                ff05e364-6aea-4915-8098-b2250bf6631a
                © 2020 Brady et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 14 July 2020
                : 28 November 2020
                Page count
                Figures: 3, Tables: 3, Pages: 15
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/100013743, World Cancer Research Fund International;
                Award ID: 2012/1003
                Award Recipient :
                This research was sponsored by Cancer Trials Ireland. Author SF received funding from the World Cancer Research Fund ( https://www.wcrf.org/; grant number 2012/1003). The funders had no role in the collection of specimens or analysis of data throughout the study duration, and had no role in preparation or review of this manuscript for publication.
                Categories
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                Custom metadata
                There are ethical restrictions to the sharing of this de-identified dataset. Informed consent for use of the data does not allow for sharing individual data publicly. Trinity College Dublin is responsible for clinical data and the data protection officer can be contacted at evelyn.fox@ 123456tcd.ie for all future data requests. The contact details for each ethics committee are as follows: NRES Committee London—Camden & Islington ( nrescommittee.london-camdenislington@ 123456nhs.net ), The Mater Misericordia Hospital Research Ethics Committee, Dublin ( mmh@ 123456mater.ie ), Beaumont Hospital Ethics, Dublin ( beaumontethics@ 123456rcsi.com ), SJH/AMNCH Research Ethics Committee, Dublin ( researchethics@ 123456tuh.ie ) and St Luke’s Radiation Oncology Network, Dublin ( radiotherapy.stlukes@ 123456slh.ie ).

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