{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T23:41:26Z","timestamp":1788478886168,"version":"build-2803163510"},"reference-count":51,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,7,14]],"date-time":"2023-07-14T00:00:00Z","timestamp":1689292800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Pancreatic cancer is associated with poor prognosis, and limited treatment options are available for the majority of patients. Natural killer (NK) cells in combination with antibodies inducing antibody-dependent cell-mediated cytotoxicity (ADCC) could be a highly effective new therapeutic option in pancreatic cancer. Accurate predictive preclinical models are needed to develop successful NK cell immunotherapy. Tumor organoids, in vitro 3D organ-like structures that retain important pathophysiological characteristics of the in vivo tumor, may provide such a model. In the current study, we assessed the cytotoxic potential of adoptive NK cells against human pancreatic cancer organoids. We hypothesized that NK cell anti-tumor responses could be enhanced by including ADCC-triggering antibodies.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We performed cytotoxicity assays with healthy donor-derived IL-2-activated NK cells and pancreatic cancer organoids from four patients. A 3D cytotoxicity assay using live-cell-imaging was developed and enabled real-time assessment of the response.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>We show that NK cells migrate to and target pancreatic cancer organoids, resulting in an increased organoid death, compared to the no NK cell controls (reaching an average fold change from baseline of 2.1\u00b10.8 vs 1.4\u00b10.6). After 24-hours of co-culture, organoid 2D growth increased. Organoids from 2 out of 4 patients were sensitive to NK cells, while organoids from the other two patients were relatively resistant, indicating patient-specific heterogeneity among organoid cultures. The ADCC-inducing antibodies avelumab (anti-PD-L1) and trastuzumab (anti-HER2) increased NK cell-induced organoid cell death (reaching an average fold change from baseline of 3.5\u00b11.0 and 4.5\u00b11.8, respectively). Moreover, combination therapy with avelumab or trastuzumab resulted in complete disintegration of organoids. Finally, inclusion of ADCC-inducing antibodies was able to overcome resistance in NK-organoid combinations with low or no kill.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>These results support the use of organoids as a relevant and personalized model to study the anti-tumor response of NK cells <jats:italic>in vitro<\/jats:italic> and the potential of ADCC-inducing antibodies to enhance NK cell effector function.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2023.1133796","type":"journal-article","created":{"date-parts":[[2023,7,14]],"date-time":"2023-07-14T19:46:27Z","timestamp":1689363987000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":23,"title":["Antibody-dependent cellular cytotoxicity-inducing antibodies enhance the natural killer cell anti-cancer response against patient-derived pancreatic cancer organoids"],"prefix":"10.3389","volume":"14","author":[{"given":"Nicky A.","family":"Beelen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Merel R.","family":"Aberle","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Virginia","family":"Bruno","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven W. M.","family":"Olde Damink","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gerard M. J.","family":"Bos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sander S.","family":"Rensen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lotte","family":"Wieten","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,7,14]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.3322\/caac.21660","article-title":"Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA: Cancer J Clin"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1080\/0284186X.2016.1197419","article-title":"More deaths from pancreatic cancer than breast cancer in the EU by 2017","volume":"55","author":"Ferlay","year":"2016","journal-title":"Acta Oncol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-14-0155","article-title":"Projecting cancer incidence and deaths to 2030: the unexpected burden of thyroid, liver, and pancreas cancers in the united states","volume":"74","author":"Rahib","year":"2014","journal-title":"Cancer Res"},{"key":"B4","doi-asserted-by":"publisher","first-page":"788","DOI":"10.1016\/j.ccell.2020.08.004","article-title":"Challenges and opportunities for pancreatic cancer immunotherapy","volume":"38","author":"Bear","year":"2020","journal-title":"Cancer Cell"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1158\/2326-6066.CIR-14-0027","article-title":"Immunotherapy converts nonimmunogenic pancreatic tumors into immunogenic foci of immune regulation","volume":"2","author":"Lutz","year":"2014","journal-title":"Cancer Immunol Res"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2018.12.038","article-title":"Broadening the impact of immunotherapy to pancreatic cancer: challenges and opportunities","volume":"156","author":"Balachandran","year":"2019","journal-title":"Gastroenterology"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1038\/ni1582","article-title":"Functions of natural killer cells","volume":"9","author":"Vivier","year":"2008","journal-title":"Nat Immunol"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1111\/jgh.15265","article-title":"Peripheral natural killer cell activity is associated with poor clinical outcomes in pancreatic ductal adenocarcinoma","volume":"36","author":"Lee","year":"2021","journal-title":"J Gastroenterol Hepatology"},{"key":"B9","doi-asserted-by":"publisher","first-page":"1845424","DOI":"10.1080\/2162402X.2020.1845424","article-title":"NK cells in pancreatic cancer demonstrate impaired cytotoxicity and a regulatory IL-10 phenotype","volume":"9","author":"Marcon","year":"2020","journal-title":"Oncoimmunology"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.00496","article-title":"Defective localization with impaired tumor cytotoxicity contributes to the immune escape of NK cells in pancreatic cancer patients","volume":"10","author":"Lim","year":"2019","journal-title":"Front Immunol"},{"key":"B11","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/1479-5876-11-262","article-title":"Comprehensive analysis of the percentage of surface receptors and cytotoxic granules positive natural killer cells in patients with pancreatic cancer, gastric cancer, and colorectal cancer","volume":"11","author":"Peng","year":"2013","journal-title":"J Trans Med"},{"key":"B12","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s13045-020-01014-w","article-title":"NK cell-based cancer immunotherapy: from basic biology to clinical development","volume":"14","author":"Liu","year":"2021","journal-title":"J Hematol Oncol"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1016\/j.omto.2021.10.001","article-title":"The emerging role of off-the-shelf engineered natural killer cells in targeted cancer immunotherapy","volume":"23","author":"Fabian","year":"2021","journal-title":"Mol Therapy-Oncolytics"},{"key":"B14","doi-asserted-by":"publisher","first-page":"36","DOI":"10.1016\/j.cytogfr.2020.12.003","article-title":"Chemokine networks modulating natural killer cell trafficking to solid tumors","volume":"59","author":"Yao","year":"2021","journal-title":"Cytokine Growth factor Rev"},{"key":"B15","doi-asserted-by":"publisher","first-page":"3232","DOI":"10.3390\/cancers13133232","article-title":"ADCC-inducing antibody trastuzumab and selection of KIR-HLA ligand mismatched donors enhance the NK cell anti-breast cancer response","volume":"13","author":"Ehlers","year":"2021","journal-title":"Cancers"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1007\/s00262-018-2140-1","article-title":"Daratumumab augments alloreactive natural killer cell cytotoxicity towards CD38+ multiple myeloma cell lines in a biochemical context mimicking tumour microenvironment conditions","volume":"67","author":"Mahaweni","year":"2018","journal-title":"Cancer Immunology Immunother"},{"key":"B17","doi-asserted-by":"publisher","first-page":"3285","DOI":"10.3390\/cancers12113285","article-title":"The immune checkpoint PD-1 in natural killer cells: expression, function and targeting in tumour immunotherapy","volume":"12","author":"Quatrini","year":"2020","journal-title":"Cancers"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2005.01.041","article-title":"Herceptin: mechanisms of action and resistance","volume":"232","author":"Nahta","year":"2006","journal-title":"Cancer Lett"},{"key":"B19","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s42003-021-02417-2","article-title":"3D tumor spheroid microarray for high-throughput, high-content natural killer cell-mediated cytotoxicity","volume":"4","author":"Gopal","year":"2021","journal-title":"Commun Biol"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0200664","article-title":"Anti-CD137 monoclonal antibody enhances trastuzumab-induced, natural killer cell-mediated cytotoxicity against pancreatic cancer cell lines with low human epidermal growth factor-like receptor 2 expression","volume":"13","author":"Masu","year":"2018","journal-title":"PLoS One"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1158\/1078-0432.CCR-06-0544","article-title":"Antitumor effect of trastuzumab for pancreatic cancer with high HER-2 expression and enhancement of effect by combined therapy with gemcitabine","volume":"12","author":"Kimura","year":"2006","journal-title":"Clin Cancer Res"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-018-0409-3","article-title":"Genetic and transcriptional evolution alters cancer cell line drug response","volume":"560","author":"Ben-David","year":"2018","journal-title":"Nature"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-08-4210","article-title":"A primary xenograft model of small-cell lung cancer reveals irreversible changes in gene expression imposed by culture in vitro","volume":"69","author":"Daniel","year":"2009","journal-title":"Cancer Res"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1038\/ng.3398","article-title":"Virtual microdissection identifies distinct tumor- and stroma-specific subtypes of pancreatic ductal adenocarcinoma","volume":"47","author":"Moffitt","year":"2015","journal-title":"Nat Genet"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1002\/bjs.10726","article-title":"Patient-derived organoid models help define personalized management of gastrointestinal cancer","volume":"105","author":"Aberle","year":"2018","journal-title":"J Br Surgery"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.12.021","article-title":"Organoid models of human and mouse ductal pancreatic cancer","volume":"160","author":"Boj","year":"2015","journal-title":"Cell"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1002\/jcsm.12627","article-title":"Generation and initial characterization of novel tumour organoid models to study human pancreatic cancer-induced cachexia","volume":"11","author":"Vaes","year":"2020","journal-title":"J Cachexia Sarcopenia Muscle"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2016.097","article-title":"Culture and establishment of self-renewing human and mouse adult liver and pancreas 3D organoids and their genetic manipulation","volume":"11","author":"Broutier","year":"2016","journal-title":"Nat Protoc"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-05126-w","article-title":"RASA2 ablation in T cells boosts antigen sensitivity and long-term function","volume":"609","author":"Carnevale","year":"2022","journal-title":"Nature"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2018100928","article-title":"3D model for CAR-mediated cytotoxicity using patient-derived colorectal cancer organoids","volume":"38","author":"Schnalzger","year":"2019","journal-title":"EMBO J"},{"key":"B31","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41467-021-26059-4","article-title":"Single-cell analysis of patient-derived PDAC organoids reveals cell state heterogeneity and a conserved developmental hierarchy","volume":"12","author":"Krieger","year":"2021","journal-title":"Nat Commun"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.03.053","article-title":"Prospective derivation of a living organoid biobank of colorectal cancer patients","volume":"161","author":"Van de Wetering","year":"2015","journal-title":"Cell"},{"key":"B33","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1016\/j.ctrv.2019.06.005","article-title":"Checkpoint inhibitors in pancreatic cancer","volume":"78","author":"Henriksen","year":"2019","journal-title":"Cancer Treat Rev"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2016-311585","article-title":"IL-6 and PD-L1 antibody blockade combination therapy reduces tumour progression in murine models of pancreatic cancer","volume":"67","author":"Mace","year":"2018","journal-title":"Gut"},{"key":"B35","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s12943-018-0928-4","article-title":"Role of the tumor microenvironment in PD-L1\/PD-1-mediated tumor immune escape","volume":"18","author":"Jiang","year":"2019","journal-title":"Mol Cancer"},{"key":"B36","doi-asserted-by":"publisher","first-page":"335","DOI":"10.1016\/j.jaci.2016.04.025","article-title":"Identification of a subset of human natural killer cells expressing high levels of programmed death 1: a phenotypic and functional characterization","volume":"139","author":"Pesce","year":"2017","journal-title":"J Allergy Clin Immunol"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1158\/2159-8290.CD-18-1259","article-title":"The mechanism of anti\u2013pd-l1 antibody efficacy against pd-l1\u2013negative tumors identifies nk cells expressing pd-l1 as a cytolytic effector","volume":"9","author":"Dong","year":"2019","journal-title":"Cancer Discov"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.01179","article-title":"Killer ig-like receptors (KIRs): their role in NK cell modulation and developments leading to their clinical exploitation","volume":"10","author":"Pende","year":"2019","journal-title":"Front Immunol"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1038\/s41588-021-00889-w","article-title":"Genome-scale screens identify factors regulating tumor cell responses to natural killer cells","volume":"53","author":"Sheffer","year":"2021","journal-title":"Nat Genet"},{"key":"B40","doi-asserted-by":"publisher","first-page":"1731","DOI":"10.1016\/j.cell.2018.10.014","article-title":"Anti-NKG2A mAb is a checkpoint inhibitor that promotes anti-tumor immunity by unleashing both T and NK cells","volume":"175","author":"Andr\u00e9","year":"2018","journal-title":"Cell"},{"key":"B41","doi-asserted-by":"publisher","first-page":"1586","DOI":"10.1016\/j.cell.2018.07.009","article-title":"Generation of tumor-reactive T cells by co-culture of peripheral blood lymphocytes and tumor organoids","volume":"174","author":"Dijkstra","year":"2018","journal-title":"Cell"},{"key":"B42","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1016\/j.addr.2014.06.005","article-title":"Matrigel: from discovery and ECM mimicry to assays and models for cancer research","volume":"79","author":"Benton","year":"2014","journal-title":"Advanced Drug Delivery Rev"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.00297","article-title":"Blockade of stromal Gas6 alters cancer cell plasticity, activates NK cells, and inhibits pancreatic cancer metastasis","volume":"11","author":"Ireland","year":"2020","journal-title":"Front Immunol"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1097\/MPA.0000000000001464","article-title":"Pancreatic stellate cells promote tumor progression by promoting an immunosuppressive microenvironment in murine models of pancreatic cancer","volume":"49","author":"Li","year":"2020","journal-title":"Pancreas"},{"key":"B45","doi-asserted-by":"publisher","first-page":"6","DOI":"10.1016\/j.trecan.2015.12.003","article-title":"Microfluidics: a new tool for modeling cancer\u2013immune interactions","volume":"2","author":"Boussommier-Calleja","year":"2016","journal-title":"Trends Cancer"},{"key":"B46","doi-asserted-by":"publisher","first-page":"389","DOI":"10.1146\/annurev.immunol.26.021607.090404","article-title":"Death by a thousand cuts: granzyme pathways of programmed cell death","volume":"26","author":"Chowdhury","year":"2008","journal-title":"Annu Rev Immunol"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1038\/sj.onc.1207232","article-title":"TRAIL and apoptosis induction by TNF-family death receptors","volume":"22","author":"Wang","year":"2003","journal-title":"Oncogene"},{"key":"B48","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s40425-019-0553-9","article-title":"Cocultures of human colorectal tumor spheroids with immune cells reveal the therapeutic potential of MICA\/B and NKG2A targeting for cancer treatment","volume":"7","author":"Courau","year":"2019","journal-title":"J Immunother cancer"},{"key":"B49","doi-asserted-by":"publisher","first-page":"3483","DOI":"10.3748\/wjg.v27.i24.3483","article-title":"Natural killer cells in pancreatic cancer stroma","volume":"27","author":"Fincham","year":"2021","journal-title":"World J Gastroenterology"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.1038\/bjc.2012.18","article-title":"Multicentre phase II trial of trastuzumab and capecitabine in patients with HER2 overexpressing metastatic pancreatic cancer","volume":"106","author":"Harder","year":"2012","journal-title":"Br J Cancer"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1200694","article-title":"Safety and activity of anti\u2013PD-L1 antibody in patients with advanced cancer","volume":"366","author":"Brahmer","year":"2012","journal-title":"New Engl J Med"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2023.1133796\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,14]],"date-time":"2023-07-14T19:46:32Z","timestamp":1689363992000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2023.1133796\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,7,14]]},"references-count":51,"alternative-id":["10.3389\/fimmu.2023.1133796"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2023.1133796","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,7,14]]},"article-number":"1133796"}}