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Target Identification and Validation in Drug Discovery pp 151–162Cite as

3D-3 Tumor Models in Drug Discovery for Analysis of Immune Cell Infiltration

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1953))

Abstract

The cross talk between tumor cells and other cells present in the tumor microenvironment such as stromal and immune cells highly influences the behavior and progression of disease. Understanding the underlying mechanisms of interaction is a prerequisite to develop new treatment strategies and to prevent or at least reduce therapy failure in the future. Specific reactivation of the patient’s immune system is one of the major goals today. However, standard two-dimensional (2D) cell culture techniques lack the necessary complexity to address related questions. Novel three-dimensional (3D) in vitro models—embedded in a matrix or encapsulated in alginate—recapitulate the in vivo situation much better. Cross talk between different cell types can be studied starting from co-cultures. As cancer immune modulation is becoming a major research topic, 3D in vitro models represent an important tool to address immune regulatory/modulatory questions for T, NK, and other cells of the immune system. The 3D systems consisting of tumor cells, fibroblasts, and immune cells (3D-3) already proved as a reliable tool for us. For instance, we made use of those models to study the molecular mechanisms of the cross talk of non-small cell lung cancer (NSCLC) and fibroblasts, to unveil macrophage plasticity in the tumor microenvironment and to mirror drug responses in vivo. Generation of those 3D models and how to use them to study immune cell infiltration and activation will be described in the present book chapter.

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Abbreviations

3D:

Three-dimensional

CAFs:

Cancer-associated fibroblasts

CFSE:

Carboxyfluorescein succinimidyl ester

CMAC:

7-Amino-4-chloromethylcoumarin

CTLA4:

Cytotoxic T-lymphocyte-associated protein 4 (CD152)

DMEM:

Dulbecco’s Modified Eagle’s Medium

DO:

Dissolved oxygen

ECM:

Extracellular matrix

EDTA:

Ethylenediaminetetraacetic acid

EMT:

Epithelial-mesenchymal transition

FBS:

Fetal bovine serum

FCS:

Fetal calf serum

GFP:

Green fluorescence protein

HDFs:

Human dermal fibroblasts

HEPES:

4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid

LAG3:

Lymphocyte activation gene 3 (CD223)

MAPK:

Mitogen-activated protein kinase

mTOR:

Mammalian target of rapamycin

NSCLC:

Non-small cell lung cancer

PBS:

Phosphate-buffered saline

PD1:

Programmed cell death protein 1

PNKs:

Primary NK cells

RFP:

Red fluorescence protein

RPMI medium:

Roswell Park Memorial Institute medium

RT:

Room temperature

TAM:

Tumor-associated macrophage

TGF:

Tumor growth factor

TIM3:

T-cell membrane protein 3 (CD366)

T regs:

Regulatory T cells

ZO-1:

Zonula occludens, tight junction protein

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Acknowledgments

We thank Nathalie Harrer for sharing her expertise on cultivation of immune cells and Martha Majewska for the technical support.

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Authors and Affiliations

  1. Boehringer Ingelheim RCV GmbH & Co KG, Vienna, Austria

    Annika Osswald, Viola Hedrich & Wolfgang Sommergruber

Authors
  1. Annika Osswald
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  2. Viola Hedrich
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  3. Wolfgang Sommergruber
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Corresponding author

Correspondence to Wolfgang Sommergruber .

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Editors and Affiliations

  1. Pharmacology and Translational Research, Boehringer Ingelheim RCV GmbH & Co KG, Vienna, Austria

    Jürgen Moll

  2. Department of Cancer Cell Signaling, Boehringer Ingelheim RCV GmBH & Co KG, Vienna, Austria

    Sebastian Carotta

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Osswald, A., Hedrich, V., Sommergruber, W. (2019). 3D-3 Tumor Models in Drug Discovery for Analysis of Immune Cell Infiltration. In: Moll, J., Carotta, S. (eds) Target Identification and Validation in Drug Discovery. Methods in Molecular Biology, vol 1953. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9145-7_10

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  • DOI: https://doi.org/10.1007/978-1-4939-9145-7_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9144-0

  • Online ISBN: 978-1-4939-9145-7

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