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Review
. 2020 Dec 23;24(1):101985.
doi: 10.1016/j.isci.2020.101985. eCollection 2021 Jan 22.

Engineering approaches for studying immune-tumor cell interactions and immunotherapy

Sarah E Shelton  1   2 , Huu Tuan Nguyen  1 , David A Barbie  2   3 , Roger D Kamm  1   4
Affiliations
Review

Engineering approaches for studying immune-tumor cell interactions and immunotherapy

Sarah E Shelton et al. iScience. .

Abstract

This review describes recent research that has advanced our understanding of the role of immune cells in the tumor microenvironment (TME) using advanced 3D in vitro models and engineering approaches. The TME can hinder effective eradication of tumor cells by the immune system, but immunotherapy has been able to reverse this effect in some cases. However, patient-to-patient variability in response suggests that we require deeper understanding of the mechanistic interactions between immune and tumor cells to improve response and develop novel therapeutics. Reconstruction of the TME using engineered 3D models allows high-resolution observation of cell interactions while allowing control of conditions such as hypoxia, matrix stiffness, and flow. Moreover, patient-derived organotypic models are an emerging tool for prediction of drug efficacy. This review highlights the importance of modeling and understanding the immune TME and describes new tools for identifying new biological targets, drug testing, and strategies for personalized medicine.

Keywords: Bioengineering; Cancer; Components of the Immune System.

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Conflict of interest statement

RDK is a co-founder and has a significant financial interest in AIM Biotech, a company that manufactures microfluidic systems and has received research funding from Elstar, 10.13039/100002429Amgen and 10.13039/100005614Biogen. DAB is an inventor on patents related on manipulating, culturing, and evaluating tumor spheroids. DAB is a consultant for N of One and Tango Therapeutics, has received honoraria from Loxo Oncology and Madalon Consulting, research grants from 10.13039/100002491BMS, 10.13039/100004336Novartis, 10.13039/100004312Lilly, and 10.13039/100005564Gilead Sciences, and is a co-founder and on the scientific advisory board of Xsphera Biosciences Inc.

Figures

None
Graphical abstract
Figure 1
Figure 1
Infiltrating immune cells and physical conditions regulate several cellular events during tumor development and dissemination Bottom left box: anti-tumoral leukocytes such as cytotoxic T lymphocytes (CTL), conventional dendritic cells (cDC), natural killer (NK) cells, type 1 helper T cell (Th1), and cytotoxic macrophage (cMΦ). Anti-tumoral immune cells infiltrate to the tumor and kill tumor cells by different mechanisms: anti-tumoral cytokine secretion, phagocytosis, or tumor-specific cytotoxicity. Bottom right box: pro-tumoral cells such as regulatory T cells (T reg), tumor-associated macrophages (TAMs), myeloid-derived suppressor cells (MDSCs), and tumor-associated neutrophils (TANs) secrete pro-tumoral cytokines that promote tumor cell survival and proliferation and induce angiogenesis or lymphangiogenesis. They also support tumor cell metastasis and suppress anti-tumoral immune cell recruitment and function in both primary and metastatic sites. Physical conditions within the tumor such as ECM stiffness, low pH, and additional factors such as hypoxia and high interstitial fluid pressure also tend to suppress the recruitment and function of anti-tumoral leukocytes.
Figure 2
Figure 2
Adaptive immunity in cancer focuses on the role of T lymphocytes in the TME (A and B) Immunohistochemical staining of CD8+ lymphocytes in colorectal cancer illustrating examples of “immune excluded” (panel A) versus “immune infiltrated” (panel B) tumors. (C) Kaplan-Meier curve revealing the association between survival and the number of infiltrating lymphocytes observed. (D–F) Three-dimensional, in vitro approaches to studying tumor-immune interactions including organoid culture in multiwall plates with lymphocytes in suspension (D), microfluidic devices with tumor and lymphocyte compartments separated by microchannels (E), and microfluidic devices with tumor organoids embedded in hydrogel channels immediately adjacent to a lymphocyte compartment (F). (G) Studies focusing on the trajectory and activity of T cells in the cancer microenvironment, including chemoattractant gradients to recruit T cells, T cell adhesion and extravasation through the vascular endothelium, migration through extracellular matrix, and interaction with cancer cells resulting in the secretion of proteins such as IFN-γ, perforin, and granzymes, culminating tumor cell killing. Panels A–C adapted from (Chiba et al., 2004).
Figure 3
Figure 3
Three-dimensional models employed to study different cellular events during tumor progression and dissemination (A) Immune cell recruitment. Microfluidic device with two adjacent gel channels (red) flanked by media channels (blue) is used for the study of macrophage migration in the presence of tumor cells. Tumor cells and macrophages are suspended in collagen I in separate gel channels. Single-cell analysis of macrophage migration demonstrates that macrophages have higher speed and directedness when co-cultured with tumor cells compared with controls. (B) Tumor cell dissemination. A device similar to that in (A) was used to study epithelial-mesenchymal transition (EMT). Tumor spheroids are suspended in gel, and macrophages are either suspended together in the same or an adjacent but different channel than the tumor spheroid. After 36 h, tumor spheroids cultured with M2a macrophages, but no other sub-type, dispersed more readily when in contact with the tumor as opposed to separated. Scale bars: 100 μm. (C) Tumor cell invasion at the metastatic site. First, monocytes are introduced into the endothelialized center channel and allowed to extravasate. Two days later, tumor cells are introduced into the same channel. Monocyte migration created microtracks that facilitated tumor cell invasion into the ECM. Scale bars:10 µm. (D) Angiogenesis. Open-top, stackable, microfluidic devices demonstrate different effects of tumor cell phenotypes on angiogenesis through induction of TAMs. This study showed that different tumor prostate cancer phenotypes resulted in different vessel morphologies through modulation of TAM phenotypes. Scale bars: 150 μm. (E) Cytotoxicity. Effects of antibody drugs on tumor cell killing by NK cells is recapitulated by a microfluidic platform that has tumor cells and NK cells in gel suspension, and drugs diffuse from the media channel. This study reports the capability of NK cells to induce tumor cell apoptosis. (F) Immunosuppression. Effector T cell suppression by monocytic cells are modeled by capturing tumor-killing efficacy within a microfluidic device. Tumor cells and monocytes are suspended in the gel channel, whereas effector T cells are introduced into the media channel. T cells migrate then to the gel channel, toward tumor cells and kill them. However, the presence of monocytic cells in the gel impeded the killing efficacy. Using check point blockade targeting PD-L1/PD-1 signaling, T cell cytotoxicity is restored. Figures are adapted from the following publications: (A–F) panels are respectively from Lee et al., 2020; Bai et al., 2015; Kim et al., 2019a; Yu et al., 2019; Ayuso et al., 2019; Lee et al., 2018):
Figure 4
Figure 4
Approaches and characterization of patient-derived organotypic models in culture (A) Common methods for patient-derived organoid culture include the air-liquid interface in which organoids are cultured in collagen and medium diffuses through a permeable support or the approach where organoids are embedded in Matrigel and surrounded by cell culture medium. (B) Histology sections comparing matched biopsies and patient-derived organoids from colon and lung adenocarcinomas. (C) Stereomicroscopy images of patient-derived colon adenocarcinoma organoids at day 2 and 30 of culture and after passaging. (D) Phase contrast images of murine-derived organotypic tumor spheroids (MDOTS) embedded in a collagen gel compartment of a microfluidic device. (E) Fluorescent staining of CD8 T cells and CD45 immune population within a murine organotypic model of colorectal cancer. (F) In vitro culture permits repeated media sampling for cytokine profiling throughout several days of culture to identify changes through time or in response to treatment. Panels (B and C) adapted from (Neal et al., 2018) and (D–F) from (Jenkins et al., 2018).
See this image and copyright information in PMC

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