Tumor-immune-interaction in a pancreatic cancer organoid co-culture model

Zweck: In Anbetracht der immer noch sehr schlechten Prognose und weiter steigenden Inzidenz des duktalen Adenokarzinoms des Pankreas (PDAC) sind klinische translationale Studien zur Früherkennung, Behandlung und Prognose von entscheidender Bedeutung. In dieser Studie untersuchten wir die Wechselwirkungen von Tumor- und Immunzellen, indem wir ein Kokulturmodell primärer menschlicher PDAC-Organoide und peripherer mononukleärer Blutzellen (PBMCs) etablierten, die von denselben Patienten stammten. Methoden: Wir haben eine Kokulturmethode für von Patienten stammende Organoide und passende PBMCs bestimmt, um die Differenzierung von Gedächtnis-T-Zell-Subtypen und regulatorischen T-Zellen (Tregs) durch Fluoreszenz-aktivierten Zellsortierer (FACS) zu testen. Ergebnisse: Die Ergebnisse waren bei 4 Patienten-Co-Kulturen unterschiedlich, wobei die Co-Kultur eines Patienten einen Anstieg der CD4+ Tcm- und Tnaiv-, CD8+ Tcm- und Tnaiv-Zellpopulationen und einen Rückgang der CD4+ Tem- und Teff-, CD8+ Tem-Zellen, verglichen mit denen in der Gruppe der allein kultivierten PBMCs; und mit einer Kokultur eines anderen Patienten, die eine erhöhte Anzahl von CD4+ Tnaiv- und CD8+ Tcm-Zellen zeigt, während eine verringerte Anzahl von CD4+ Tem- und CD8+ Tem-Zellen. Abgesehen davon war die Population von Treg-Zellen in allen Co-Kulturen der PDAC-Patienten höher als in ihren gematchten Kontrollen. Schlussfolgerung: Hier wurde ein autologes Kokulturmodell mit von Patienten stammenden Organoiden und PBMCs etabliert, um die Wechselwirkung zwischen Krebszellen und verschiedenen Immunzellen bei einzelnen PDAC-Patienten zu untersuchen. Mit diesem Modell steht uns ein Werkzeug zur Verfügung, mit dem wir die Wirksamkeit von Immuntherapien vorhersagen könnten und so dazu beitragen können, das Outcome unserer Patienten zu verbessern.Purpose: Considering the rising incidence and dismal prognosis of pancreatic ductal adenocarcinoma (PDAC), clinical translational studies for early diagnosis, treatment, and prognosis are of crucial importance. In this project, we studied interactions of tumor and immune cells by establishing a co-culture model of primary human PDAC organoids and peripheral blood mononuclear cells (PBMCs) derived from the same patients. Methods: We determined a co-culture method for patient-derived organoids and matched PBMCs to test the differentiation of Memory T cell subtypes and Regulatory T cells (Tregs) by Fluorescence Activated Cell Sorter (FACS). Results: The results were variable in 4 patient co-cultures, with one patient’s co-culture showing an increase in CD4+ Tcm- and Tnaiv-, CD8+ Tcm- and Tnaiv cell populations, and a decrease in CD4+ Tem- and Teff-, CD8+ Tem cells, compared to those in PBMCs cultured alone group; and with the co-culture from another patient displaying elevated numbers of CD4+ Tnaiv- and CD8+ Tcm cells, whereas reduced numbers of CD4+ Tem- and CD8+ Tem cells. Besides that, the population of Treg cells in all co-cultures of the PDAC patients was higher than in their matched controls. Conclusion: Here an autologous co-culture model with patient-derived organoids and PBMCs was established to study the crosstalk between cancerous cells and various immune cells in individual PDAC patients. Having this model available we have a tool at hand which might predict the effectiveness of immune therapies thereby helping to improve the outcome of our patients

Organoids Models for the Study of Cell-Cell Interactions

Organoids have arisen as promising model systems in biomedical research and regenerative medicine due to their potential to reproduce the original tissue architecture and function. In the research field of cell–cell interactions, organoids mimic interactions taking place during organogenesis, including the processes that conduct to multi-lineage differentiation and morphogenetic processes, during immunology response and disease development and expansion. This chapter will address the basis of organoids origin, their importance on immune system cell–cell interactions and the benefits of using them in biomedicine, specifically their potential applications in regenerative medicine and personalized therapy. Organoids might represent a personalized tool for patients to receive earlier diagnoses, risk assessments, and more efficient treatments

Progress and potential in organoid research

Tissue and organ biology are very challenging to study in mammals, and progress can be hindered, particularly in humans, by sample accessibility and ethical concerns. However, advances in stem cell culture have made it possible to derive in vitro 3D tissues called organoids, which capture some of the key multicellular, anatomical and even functional hallmarks of real organs at the micrometre to millimetre scale. Recent studies have demonstrated that organoids can be used to model organ development and disease and have a wide range of applications in basic research, drug discovery and regenerative medicine. Researchers are now beginning to take inspiration from other fields, such as bioengineering, to generate organoids that are more physiologically relevant and more amenable to real-life applications

Melanoma Models for the Next Generation of Therapies

There is a lack of appropriate melanoma models that can be used to evaluate the efficacy of novel therapeutic modalities. Here, we discuss the current state of the art of melanoma models including genetically engineered mouse, patient-derived xenograft, zebrafish, and ex vivo and in vitro models. We also identify five major challenges that can be addressed using such models, including metastasis and tumor dormancy, drug resistance, the melanoma immune response, and the impact of aging and environmental exposures on melanoma progression and drug resistance. Additionally, we discuss the opportunity for building models for rare subtypes of melanomas, which represent an unmet critical need. Finally, we identify key recommendations for melanoma models that may improve accuracy of preclinical testing and predict efficacy in clinical trials, to help usher in the next generation of melanoma therapies