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      PD-L1 expressing circulating tumour cells in head and neck cancers

      case-report

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          Abstract

          Background

          Blockade of the PD-1/PD-L1 immune checkpoint pathway is emerging as a promising immunotherapeutic approach for the management and treatment of head and neck cancer patients who do not respond to 1st/2nd line therapy. However, as checkpoint inhibitors are cost intensive, identifying patients who would most likely benefit from anti PD-L1 therapy is required. Developing a non-invasive technique would be of major benefit to the patient and to the health care system.

          Case presentation

          We report the case of a 56 year old man affected by a supraglottic squamous cell carcinoma (SCC). A CT scan showed a 20 mm right jugulodigastric node and suspicious lung lesions. The lung lesion was biopsied and confirmed to be consistent with SCC. The patient was offered palliative chemotherapy. At the time of presentation, a blood sample was taken for circulating tumour cell (CTC) analysis. The dissemination of cancer was confirmed by the detection of CTCs in the peripheral blood of the patient, measured by the CellSearch System (Janssen Diagnostics). Using marker-independent, low-shear spiral microfluidic technology combined with immunocytochemistry, CTC clusters were found in this patient at the same time point, expressing PD-L1.

          Conclusion

          This report highlights the potential use of CTCs to identify patients which might respond to anti PD-L1 therapy.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12885-017-3316-3) contains supplementary material, which is available to authorized users.

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          Most cited references16

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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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            Slanted spiral microfluidics for the ultra-fast, label-free isolation of circulating tumor cells.

            The enumeration and characterization of circulating tumor cells (CTCs), found in the peripheral blood of cancer patients, provide a potentially accessible source for cancer diagnosis and prognosis. This work reports on a novel spiral microfluidic device with a trapezoidal cross-section for ultra-fast, label-free enrichment of CTCs from clinically relevant blood volumes. The technique utilizes the inherent Dean vortex flows present in curvilinear microchannels under continuous flow, along with inertial lift forces which focus larger CTCs against the inner wall. Using a trapezoidal cross-section as opposed to a traditional rectangular cross-section, the position of the Dean vortex core can be altered to achieve separation. Smaller hematologic components are trapped in the Dean vortices skewed towards the outer channel walls and eventually removed at the outer outlet, while the larger CTCs equilibrate near the inner channel wall and are collected from the inner outlet. By using a single spiral microchannel with one inlet and two outlets, we have successfully isolated and recovered more than 80% of the tested cancer cell line cells (MCF-7, T24 and MDA-MB-231) spiked in 7.5 mL of blood within 8 min with extremely high purity (400-680 WBCs mL(-1); ~4 log depletion of WBCs). Putative CTCs were detected and isolated from 100% of the patient samples (n = 10) with advanced stage metastatic breast and lung cancer using standard biomarkers (CK, CD45 and DAPI) with the frequencies ranging from 3-125 CTCs mL(-1). We expect this simple and elegant approach can surmount the shortcomings of traditional affinity-based CTC isolation techniques as well as enable fundamental studies on CTCs to guide treatment and enhance patient care.
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              Ultra-fast, label-free isolation of circulating tumor cells from blood using spiral microfluidics.

              Circulating tumor cells (CTCs) are rare cancer cells that are shed from primary or metastatic tumors into the peripheral blood circulation. Phenotypic and genetic characterization of these rare cells can provide important information to guide cancer staging and treatment, and thus further research into their characteristics and properties is an area of considerable interest. In this protocol, we describe detailed procedures for the production and use of a label-free spiral microfluidic device to allow size-based isolation of viable CTCs using hydrodynamic forces that are present in curvilinear microchannels. This spiral system enables us to achieve ≥ 85% recovery of spiked cells across multiple cancer cell lines and 99.99% depletion of white blood cells in whole blood. The described spiral microfluidic devices can be produced at an extremely low cost using standard microfabrication and soft lithography techniques (2-3 d), and they can be operated using two syringe pumps for lysed blood samples (7.5 ml in 12.5 min for a three-layered multiplexed chip). The fast processing time and the ability to collect CTCs from a large patient blood volume allows this technique to be used experimentally in a broad range of potential genomic and transcriptomic applications.
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                Author and article information

                Contributors
                arutha.kulasinghe@qut.edu.au
                reception@brisbaneent.com.au
                lizkenny@bigpond.net.au
                m.warkiani@undw.edu.au
                colleen.nelson@qut.edu.au
                +61 7 3138 0830 , chamindie.punyadeera@qut.edu.au
                Journal
                BMC Cancer
                BMC Cancer
                BMC Cancer
                BioMed Central (London )
                1471-2407
                16 May 2017
                16 May 2017
                2017
                : 17
                : 333
                Affiliations
                [1 ]ISNI 0000000089150953, GRID grid.1024.7, The School of Biomedical Sciences, Institute of Health and Biomedical Innovation, , Queensland University of Technology, ; Kelvin Grove, QLD Australia
                [2 ]Translational Research Institute, Brisbane, Australia
                [3 ]ISNI 0000 0004 0380 2017, GRID grid.412744.0, Department of Otolaryngology, , Princess Alexandra Hospital, ; QLD, Woolloongabba, Australia
                [4 ]ISNI 0000 0000 9320 7537, GRID grid.1003.2, School of Medicine, , University of Queensland, ; Brisbane, QLD Australia
                [5 ]ISNI 0000 0001 0688 4634, GRID grid.416100.2, , Royal Brisbane and Women’s Hospital, ; Brisbane, QLD Australia
                [6 ]ISNI 0000 0004 0380 0804, GRID grid.415606.0, Central Integrated Regional Cancer Service, , Queensland Health, ; Brisbane, QLD Australia
                [7 ]ISNI 0000 0004 4902 0432, GRID grid.1005.4, School of Mechanical and Manufacturing Engineering, Australian Centre for NanoMedicine, , University of New South Wales, ; Sydney, Australia
                [8 ]Garvan Institute for Biomedical Research, Sydney, Australia
                [9 ]ISNI 0000 0004 0389 4302, GRID grid.1038.a, School of Medical Sciences, , Edith Cowan University, ; Joondalup, Perth, WA 6027 Australia
                [10 ]ISNI 0000 0004 0380 2017, GRID grid.412744.0, Australian Prostate Cancer Research Centre - Queensland, Institute of Health and Biomedical Innovation, , Queensland University of Technology, Princess Alexandra Hospital, Translational Research Institute Brisbane, ; Brisbane, Australia
                Author information
                http://orcid.org/0000-0003-3224-7350
                Article
                3316
                10.1186/s12885-017-3316-3
                5434641
                28511705
                064fe33f-4201-47be-9c4f-eda15efdf5e6
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 12 October 2016
                : 2 May 2017
                Categories
                Case Report
                Custom metadata
                © The Author(s) 2017

                Oncology & Radiotherapy
                pd-l1,head and neck cancers,circulating tumour cells,non-invasive tools,liquid biopsy

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