Antibody-dependent cellular cytotoxicity-inducing antibodies enhance the natural killer cell anti-cancer response against patient-derived pancreatic cancer organoids

Abstract

IntroductionPancreatic 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.MethodsWe 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.ResultsWe 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±0.8 vs 1.4±0.6). 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±1.0 and 4.5±1.8, 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.DiscussionThese results support the use of organoids as a relevant and personalized model to study the anti-tumor response of NK cells in vitro and the potential of ADCC-inducing antibodies to enhance NK cell effector function.

The Prospect of Harnessing the Microbiome to Improve Immunotherapeutic Response in Pancreatic Cancer

“…Beelen [25] ICI Increased NK-cell-induced PDAC organoid cell death with anti-PD-L1 or anti-HER2 therapy.…”

Microfluidics: a concise review of the history, principles, design, applications, and future outlook

“…227 This showed that stem cells can self-organize into organoids that resemble native organs. At present, researchers can cultivate organoids derived from stem cells that mimic the physiological attributes of diverse human organs like blood vessels, 228 kidneys, 229,230 pancreas, 231 lungs, 232,233 intestine 234 and brain. 235,236 These replicas find applications in emulating genetic disorders, advancing personalized medicine, conducting drug tests, and investigating the roots of diseases.…”

The Role of Chemokines in Orchestrating the Immune Response to Pancreatic Ductal Adenocarcinoma

“…While ADCC-promoting antibodies, such as trastuzumab, are FDA-approved and have been shown to induce durable anti-tumor responses in solid tumors, their efficacy in PDAC is limited [218]. However, Beelen et al demonstrate that the treatment of 3D patient-derived PDAC organoids with either trastuzumab or avelumab (an anti-PD-L1 ADCC-inducing antibody) enhances direct NK cell cytotoxicity, resulting in the destruction of the organoid [219]. Critically, the level of organoid apoptosis was dependent on the effector-to-target ratio, with more NK cells leading to more lysis [219].…”

“…However, Beelen et al demonstrate that the treatment of 3D patient-derived PDAC organoids with either trastuzumab or avelumab (an anti-PD-L1 ADCC-inducing antibody) enhances direct NK cell cytotoxicity, resulting in the destruction of the organoid [219]. Critically, the level of organoid apoptosis was dependent on the effector-to-target ratio, with more NK cells leading to more lysis [219]. This suggests that increasing intra-tumoral NK cell accumulation through chemokine gradients could increase the efficacy of ADCC-promoting antibodies targeting PDAC tumor cell antigens.…”