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      Fine-Needle Aspiration-Based Patient-Derived Cancer Organoids

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          Summary

          Patient-derived cancer organoids hold great potential to accurately model and predict therapeutic responses. Efficient organoid isolation methods that minimize post-collection manipulation of tissues would improve adaptability, accuracy, and applicability to both experimental and real-time clinical settings. Here we present a simple and minimally invasive fine-needle aspiration (FNA)-based organoid culture technique using a variety of tumor types including gastrointestinal, thyroid, melanoma, and kidney. This method isolates organoids directly from patients at the bedside or from resected tissues, requiring minimal tissue processing while preserving the histologic growth patterns and infiltrating immune cells. Finally, we illustrate diverse downstream applications of this technique including in vitro high-throughput chemotherapeutic screens, in situ immune cell characterization, and in vivo patient-derived xenografts. Thus, routine clinical FNA-based collection techniques represent an unappreciated substantial source of material that can be exploited to generate tumor organoids from a variety of tumor types for both discovery and clinical applications.

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          Highlights

          • Fine-needle aspiration (FNA) is safe, minimally invasive, and widely used clinically

          • FNA is a source of material for organoid culture and personalized medicine

          • This technique requires minimal processing, preserving histology, and immune cells

          • Downstream applications: high-throughput screens, immune analysis, and xenografts

          Abstract

          Clinical Medicine; Tissue Engineering; Cancer

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          Most cited references23

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          Generation of Tumor-Reactive T Cells by Co-culture of Peripheral Blood Lymphocytes and Tumor Organoids

          Cancer immunotherapies have shown substantial clinical activity for a subset of patients with epithelial cancers. Still, technological platforms to study cancer T-cell interactions for individual patients and understand determinants of responsiveness are presently lacking. Here, we establish and validate a platform to induce and analyze tumor-specific T cell responses to epithelial cancers in a personalized manner. We demonstrate that co-cultures of autologous tumor organoids and peripheral blood lymphocytes can be used to enrich tumor-reactive T cells from peripheral blood of patients with mismatch repair-deficient colorectal cancer and non-small-cell lung cancer. Furthermore, we demonstrate that these T cells can be used to assess the efficiency of killing of matched tumor organoids. This platform provides an unbiased strategy for the isolation of tumor-reactive T cells and provides a means by which to assess the sensitivity of tumor cells to T cell-mediated attack at the level of the individual patient.
            • Record: found
            • Abstract: not found
            • Article: not found

            Human Pancreatic Tumor Organoids Reveal Loss of Stem Cell Niche Factor Dependence during Disease Progression

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              Is Open Access

              Development of primary human pancreatic cancer organoids, matched stromal and immune cells and 3D tumor microenvironment models

              Background Patient-derived tumor models are the new standard for pre-clinical drug testing and biomarker discovery. However, the emerging technology of primary pancreatic cancer organoids has not yet been broadly implemented in research, and complex organotypic models using organoids in co-culture with stromal and immune cellular components of the tumor have yet to be established. In this study, our objective was to develop and characterize pancreatic cancer organoids and multi-cell type organotypic co-culture models to demonstrate their applicability to the study of pancreatic cancer. Methods We employed organoid culture methods and flow cytometric, cytologic, immunofluorescent and immunohistochemical methods to develop and characterize patient-derived pancreatic cancer organoids and multi-cell type organotypic co-culture models of the tumor microenvironment. Results We describe the culture and characterization of human pancreatic cancer organoids from resection, ascites and rapid autopsy sources and the derivation of adherent tumor cell monocultures and tumor-associated fibroblasts from these sources. Primary human organoids displayed tumor-like cellular morphology, tissue architecture and polarity in contrast to cell line spheroids, which formed homogenous, non-lumen forming spheres. Importantly, we demonstrate the construction of complex organotypic models of tumor, stromal and immune components of the tumor microenvironment. Activation of myofibroblast-like cancer associated fibroblasts and tumor-dependent lymphocyte infiltration were observed in these models. Conclusions These studies provide the first report of novel and disease-relevant 3D in-vitro models representing pancreatic tumor, stromal and immune components using primary organoid co-cultures representative of the tumor-microenvironment. These models promise to facilitate the study of tumor-stroma and tumor-immune interaction and may be valuable for the assessment of immunotherapeutics such as checkpoint inhibitors in the context of T-cell infiltration.

                Author and article information

                Contributors
                Journal
                iScience
                iScience
                iScience
                Elsevier
                2589-0042
                24 July 2020
                21 August 2020
                24 July 2020
                : 23
                : 8
                : 101408
                Affiliations
                [1 ]Department of Pathology, The Ohio State University, Columbus, OH 43210, USA
                [2 ]Department of Pharmacology, Vanderbilt University, Nashville, TN 37232, USA
                [3 ]Department of Biochemistry, Vanderbilt University, Nashville, TN 37232, USA
                [4 ]Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232, USA
                [5 ]Vanderbilt Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, TN 37232, USA
                [6 ]Department of Surgery, Vanderbilt University Medical Center, Nashville, TN 37232, USA
                [7 ]Vanderbilt Institute of Chemical Biology - High-Throughput Screening Facility, Vanderbilt University, Nashville, TN 37232, USA
                [8 ]Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232, USA
                [9 ]Department of Cell and Developmental Biology, Vanderbilt University, Nashville, TN 37232, USA
                [10 ]Epithelial Biology Center, Vanderbilt University Medical Center, Nashville, TN 37232, USA
                Author notes
                []Corresponding author vivian.l.weiss@ 123456vumc.org
                [11]

                Lead Contact

                Article
                S2589-0042(20)30598-8 101408
                10.1016/j.isci.2020.101408
                7415927
                32771978
                261883b9-110a-4cbe-b841-1a486ba750d4
                © 2020 The Authors

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 17 April 2020
                : 25 June 2020
                : 22 July 2020
                Categories
                Article

                clinical medicine,tissue engineering,cancer
                clinical medicine, tissue engineering, cancer

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