Abstract
The interaction between the receptor, programmed cell death protein 1 (PD-1) and ligand, programmed cell death 1 (PD-L1) is known to inhibit CD8+ cytotoxic T lymphocyte proliferation, survival, and effector function. The result of this interaction leads to evasion of immune surveillance by tumors and subsequently cancer cell proliferation. Immunotherapy via PD-L1 blockade is used for a variety of malignancies, yet the prognostic value of immune checkpoint inhibition for the treatment of gastric cancer remains controversial. Thus, preclinical models that would predict the efficacy of such therapy in a subgroup of gastric cancer patients would be an advancement in the personalized treatment of this disease. Three-dimensional organoid cultures have not only been used to investigate the mechanisms regulating development and disease, but have also been used for high-throughput drug screening for targeted personalized therapy. Here we present the methodology for the co-culture of mouse-derived gastric cancer organoids with autologous immune cells specifically for the study of PD-L1/PD-1 interactions within the tumor microenvironment in vitro.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Blanca PM, Epplein M, Correa P (2010) Gastric cancer: an infectious disease. Infect Dis Clin N Am 24(4):853–869
Neugut AI, Hayek M, Howe G (1996) Epidemiology of gastric cancer. Semin Oncol 23(3):281–291
Rebeca S, Naishadham D, Jemal A (2013) Cancer statistics. CA Cancer J Clin 63:11–30
Ahmad SA, Xia BT, Bailey CE, Abbott DE, Helmink BA, Daly MC, Thota R et al (2016) An update on gastric cancer. Curr Probl Surg 53(10):449–490
Ahmadzadeh M, Johnson LA, Heemskerk B, Wunderlich JR, Dudley ME, White DE, Rosenberg SA (2009) Tumor antigen-specific CD8 T cells infiltrating the tumor express high levels of PD-1 and are functionally impaired. Blood 114(8):1537–1544
Chen L, Gibbons DL, Goswami S, Cortez MA, Ahn Y-H, Byers LA, Zhang X et al (2014) Metastasis is regulated via microRNA-200/ZEB1 axis control of tumor cell PD-L1 expression and intratumoral immunosuppression. Nat Commun 5:5241
Reissfelder C, Stamova S, Gossmann C, Braun M, Bonertz A, Walliczek U, Grimm M, Rahbari NN, Koch M, Saadati M, Benner A (2015) Tumor-specific cytotoxic T lymphocyte activity determines colorectal cancer patient prognosis. J Clin Invest 125(2):739
Davies M (2014) New modalities of cancer treatment for NSCLC: focus on immunotherapy. Cancer Manag Res 6:63
Kyi C, Postow MA (2014) Checkpoint blocking antibodies in cancer immunotherapy. FEBS Lett 588(2):368–376
Raufi AG, Klempner SJ (2015) Immunotherapy for advanced gastric and esophageal cancer: preclinical rationale and ongoing clinical investigations. J Gastrointest Oncol 6(5):561
Hou J, Zhong Y, Xiang R, Li C, Wang L, Chen S, Li Q, Chen M, Wang L (2014) Correlation between infiltration of FOXP3+ regulatory T cells and expression of B7-H1 in the tumor tissues of gastric cancer. Exp Mol Pathol 96(3):284–291
Wu C, Zhu Y, Jiang J, Zhao J, Zhang X-G, Ning X (2006) Immunohistochemical localization of programmed death-1 ligand-1 (PD-L1) in gastric carcinoma and its clinical significance. Acta Histochem 108(1):19–24
Qing Y, Li Q, Ren T, Xia W, Peng Y, Liu G-L, Luo H et al (2015) Upregulation of PD-L1 and APE1 is associated with tumorigenesis and poor prognosis of gastric cancer. Drug Des Devel Ther 9:901
Postow MA, Callahan MK, Wolchok JD (2015) Immune checkpoint blockade in cancer therapy. J Clin Oncol 33(17):1974–1982
Schumacher MA, Aihara E, Feng R, Engevik A, Shroyer NF, Ottemann KM, Worrell RT, Montrose MH, Shivdasani RA, Zavros Y (2015) The use of murine-derived fundic organoids in studies of gastric physiology (2015). J Physiol 593(8):1809–1827
Syu LJ, Zhao X, Zhang Y, Grachtchouk M, Demitrack E, Ermilov A, Wilbert DM, Zheng X, Kaatz A, Greenson JK, Gumucio DL (2016) Invasive mouse gastric adenocarcinomas arising from Lgr5+ stem cells are dependent on crosstalk between the Hedgehog/GLI2 and mTOR pathways. Oncotarget 7(9):10255
Madaan A, Verma R, Singh AT, Jain SK, Jaggi M (2014) A stepwise procedure for isolation of murine bone marrow and generation of dendritic cells. J Biol Methods 1(1):e1. https://doi.org/10.14440/jbm.2014.12
Dorrington M (2011) Harvesting of femurs and tibia from mice. Bowdish Lab, McMaster University, Hamilton
Acknowledgements
This work is financially supported by NIH 5R01DK083402-08 (Zavros) grant.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Chakrabarti, J. et al. (2018). Mouse-Derived Gastric Organoid and Immune Cell Co-culture for the Study of the Tumor Microenvironment. In: Baratta, M. (eds) Epithelial Cell Culture. Methods in Molecular Biology, vol 1817. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8600-2_16
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8600-2_16
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8599-9
Online ISBN: 978-1-4939-8600-2
eBook Packages: Springer Protocols