bims-tuchim Biomed News
on Tumor-on-chip models
Issue of 2021‒06‒27
twelve papers selected by
Philipp Albrecht
Friedrich Schiller University
  1. High-throughput microfluidic 3D biomimetic model enabling quantitative description of the human breast tumor microenvironment.
  2. Biological gel-based microchamber array for tumor cell proliferation and migration studies in well-controlled biochemical gradients.
  3. Reconstructing the Tumor Architecture into Organoids.
  4. An integrated framework for quantifying immune-tumour interactions in a 3D co-culture model.
  5. Biophysical interactions between components of the tumor microenvironment promote metastasis.
  6. C4b-binding protein α-chain enhances antitumor immunity by facilitating the accumulation of tumor-infiltrating lymphocytes in the tumor microenvironment in pancreatic cancer.
  7. Impairing flow-mediated endothelial remodeling reduces extravasation of tumor cells.
  8. Expansion of tumor-associated Treg cells upon disruption of a CTLA-4-dependent feedback loop.
  9. Interferon-γ: teammate or opponent in the tumour microenvironment?
  10. TNF-α induces endothelial-mesenchymal transition promoting stromal development of pancreatic adenocarcinoma.
  11. Tumor Microenvironment Characteristics of Pancreatic Cancer to Determine Prognosis and Immune-Related Gene Signatures.
  12. Molecular determinants of response to PD-L1 blockade across tumor types.
  1. Acta Biomater. 2021 Jun 18. pii: S1742-7061(21)00399-8. [Epub ahead of print]
    High-throughput microfluidic 3D biomimetic model enabling quantitative description of the human breast tumor microenvironment.
    Ilana Berger Fridman, James Kostas, Michal Gregus, Somak Ray, Matthew R Sullivan, Alexander R Ivanov, Smadar Cohen, Tania Konry.
      Cancer is driven by both genetic aberrations in the tumor cells and fundamental changes in the tumor microenvironment (TME). These changes offer potential targets for novel therapeutics, yet lack of in vitro 3D models recapitulating this complex microenvironment impedes such progress. Here, we generated several tumor-stroma scaffolds reflecting the dynamic in vivo breast TME, using a high throughput microfluidic system. Alginate (Alg) or alginate-alginate sulfate (Alg/Alg-S) hydrogels were used as ECM-mimics, enabling the encapsulation and culture of tumor cells, fibroblasts and immune cells (macrophages and T cells, of the innate and adaptive immune systems, respectively). Specifically, Alg/Alg-S was shown capable of capturing and presenting growth factors and cytokines with binding affinity that is comparable to heparin. Viability and cytotoxicity were shown to strongly correlate with the dynamics of cellular milieu, as well as hydrogel type. Using on-chip immunofluorescence, production of reactive oxygen species and apoptosis were imaged and quantitatively analyzed. We then show how macrophages in our microfluidic system were shifted from a proinflammatory to an immunosuppressive phenotype when encapsulated in Alg/Alg-S, reflecting in vivo TME dynamics. LC-MS proteomic profiling of tumor cells sorted from the TME scaffolds revealed upregulation of proteins involved in cell-cell interactions and immunomodulation in Alg/Alg-S scaffolds, correlating with in vivo findings and demonstrating the appropriateness of Alg/Alg-S as an ECM biomimetic. Finally, we show the formation of large tumor-derived vesicles, formed exclusively in Alg/Alg-S scaffolds. Altogether, our system offers a robust platform for quantitative description of the breast TME that successfully recapitulates in vivo patterns. Statement of significance: Cancer progression is driven by profound changes in both tumor cells and surrounding stroma. Here, we present a high throughput microfluidic system for the generation and analysis of dynamic tumor-stroma scaffolds, that mimic the complex in vivo TME cell proportions and compositions, constructing robust in vitro models for the study of the TME. Utilizing Alg/Alg-S as a bioinspired ECM, mimicking heparin's in vivo capabilities of capturing and presenting signaling molecules, we show how Alg/Alg-S induces complex in vivo-like responses in our models. Alg/Alg-S is shown here to promote dynamic protein expression patterns, that can serve as potential therapeutic targets for breast cancer treatment. Formation of large tumor-derived vesicles, observed exclusively in the Alg/Alg-S scaffolds suggests a mechanism for tumor survival.
    Keywords:  3D disease models; biomaterials; breast cancer; microfluidics; proteomic profiling; tumor microenvironment
    DOI:  https://doi.org/10.1016/j.actbio.2021.06.025
  2. Lab Chip. 2021 Jun 23.
    Biological gel-based microchamber array for tumor cell proliferation and migration studies in well-controlled biochemical gradients.
    Jingru Yao, Guoqiang Li, Yang Jiao, Yu Zheng, Yanping Liu, Gao Wang, Lianjie Zhou, Hongfei Zhang, Xianquan Zhang, Jianwei Shuai, Qihui Fan, Fangfu Ye, Silong Lou, Guo Chen, Kena Song, Yong Liao, Liyu Liu.
      Breast cancer metastasis is a complex process controlled by multiple factors, including various cell-cell interactions, cell-environment coupling, and oxygen, nutrient and drug gradients that are intimately related to the heterogeneous breast tissue structure. In this study, we constructed a high-throughput in vitro biochip system containing an array of 642 microchambers arranged in a checkerboard configuration, with each chamber embedded in a composite extracellular matrix (ECM) composed of engineered collagen and Matrigel to mimic local heterogeneous environment in vivo. In addition, a controllable complex tetragonal chemical concentration profile can be achieved by imposing chemical compounds at the four boundaries of the chip, leading to distinct local nutrient and/or drug gradients in the individual microchambers. Here, the microchamber array with composite ECM (MACECM) device aims to simulate multiple tumor cell niches composed of both breast epithelial cells (MCF-10A-GFP) and metastatic breast cancer cells (MDA-MB-231-RFP), which enables systematic studies of cell responses to a variety of biochemical conditions. The results obtained from the MACECM studies indicate that discoidin domain receptor 1 (DDR1) inhibitor 7rh and matrix metalloproteinase inhibitor batimastat, in association with epidermal growth factor (EGF) had no significant effects on the growth of MCF-10A-GFP cells, but had significant effects on DDR1 expression and the related migratory behavior of MDA-MB-231-RFP cells. The MACECM design not only enables the construction of a more realistic in vitro model for investigating cancer cell migration mechanisms but also has considerable potential for further development as a platform for next-generation high-throughput and therapeutic screening (e.g., anti-cancer drug evaluation) and personalized medicine.
    DOI:  https://doi.org/10.1039/d0lc00951b
  3. Adv Drug Deliv Rev. 2021 Jun 18. pii: S0169-409X(21)00231-3. [Epub ahead of print] 113839
    Reconstructing the Tumor Architecture into Organoids.
    Zhimin Luo, Xingwu Zhou, Kalpana Mandal, Na He, Wally Wennerberg, Moyuan Qu, Xing Jiang, Wujin Sun, Ali Khademhosseini.
      Cancer remains one of the leading health burdens worldwide. One of the challenges hindering cancer therapy development is the substantial discrepancies between the existing cancer models and the tumor microenvironment (TME) of human patients. Constructing tumor organoids represents an emerging approach to recapitulate the pathophysiological features of the TME in vitro. Over the past decade, various approaches have been demonstrated to engineer tumor organoids as in vitro cancer models, such as incorporating multiple cellular populations, reconstructing biophysical and chemical traits, and even recapitulating structural features. In this review, we focus on engineering approaches for building tumor organoids, including biomaterial-based, microfabrication-assisted, and synthetic biology-facilitated strategies. Furthermore, we summarize the applications of engineered tumor organoids in basic cancer research, cancer drug discovery, and personalized medicine. We also discuss the challenges and future opportunities in using tumor organoids for broader applications.
    Keywords:  3D printing; Biomaterial; Genome editing; Microfluidic; Micropatterning; Tumor organoid
    DOI:  https://doi.org/10.1016/j.addr.2021.113839
  4. Commun Biol. 2021 Jun 24. 4(1): 781
    An integrated framework for quantifying immune-tumour interactions in a 3D co-culture model.
    Gheed Al-Hity, FengWei Yang, Eduard Campillo-Funollet, Andrew E Greenstein, Hazel Hunt, Myrthe Mampay, Haya Intabli, Marta Falcinelli, Anotida Madzvamuse, Chandrasekhar Venkataraman, Melanie S Flint.
      Investigational in vitro models that reflect the complexity of the interaction between the immune system and tumours are limited and difficult to establish. Herein, we present a platform to study the tumour-immune interaction using a co-culture between cancer spheroids and activated immune cells. An algorithm was developed for analysis of confocal images of the co-culture to evaluate the following quantitatively; immune cell infiltration, spheroid roundness and spheroid growth. As a proof of concept, the effect of the glucocorticoid stress hormone, cortisol was tested on 66CL4 co-culture model. Results were comparable to 66CL4 syngeneic in vivo mouse model undergoing psychological stress. Furthermore, administration of glucocorticoid receptor antagonists demonstrated the use of this model to determine the effect of treatments on the immune-tumour interplay. In conclusion, we provide a method of quantifying the interaction between the immune system and cancer, which can become a screening tool in immunotherapy design.
    DOI:  https://doi.org/10.1038/s42003-021-02296-7
  5. Biophys Rev. 2021 Jun;13(3): 339-357
    Biophysical interactions between components of the tumor microenvironment promote metastasis.
    Dimitra Vasilaki, Athina Bakopoulou, Alexandros Tsouknidas, Elaine Johnstone, Konstantinos Michalakis.
      During metastasis, tumor cells need to adapt to their dynamic microenvironment and modify their mechanical properties in response to both chemical and mechanical stimulation. Physical interactions occur between cancer cells and the surrounding matrix including cell movements and cell shape alterations through the process of mechanotransduction. The latter describes the translation of external mechanical cues into intracellular biochemical signaling. Reorganization of both the cytoskeleton and the extracellular matrix (ECM) plays a critical role in these spreading steps. Migrating tumor cells show increased motility in order to cross the tumor microenvironment, migrate through ECM and reach the bloodstream to the metastatic site. There are specific factors affecting these processes, as well as the survival of circulating tumor cells (CTC) in the blood flow until they finally invade the secondary tissue to form metastasis. This review aims to study the mechanisms of metastasis from a biomechanical perspective and investigate cell migration, with a focus on the alterations in the cytoskeleton through this journey and the effect of biologic fluids on metastasis. Understanding of the biophysical mechanisms that promote tumor metastasis may contribute successful therapeutic approaches in the fight against cancer.
    Keywords:  Actomyosin contractility; Cell mechanics; Circulating tumor cells; Cytoskeleton; Metastasis; Shear stress
    DOI:  https://doi.org/10.1007/s12551-021-00811-y
  6. J Exp Clin Cancer Res. 2021 Jun 24. 40(1): 212
    C4b-binding protein α-chain enhances antitumor immunity by facilitating the accumulation of tumor-infiltrating lymphocytes in the tumor microenvironment in pancreatic cancer.
    Kosuke Sasaki, Shigetsugu Takano, Satoshi Tomizawa, Yoji Miyahara, Katsunori Furukawa, Tsukasa Takayashiki, Satoshi Kuboki, Mamoru Takada, Masayuki Ohtsuka.
      BACKGROUND: Recent studies indicate that complement plays pivotal roles in promoting or suppressing cancer progression. We have previously identified C4b-binding protein α-chain (C4BPA) as a serum biomarker for the early detection of pancreatic ductal adenocarcinoma (PDAC). However, its mechanism of action remains unclear. Here, we elucidated the functional roles of C4BPA in PDAC cells and the tumor microenvironment.METHODS: We assessed stromal C4BPA, the C4BPA binding partner CD40, and the number of CD8+ tumor-infiltrating lymphocytes in resected human PDAC tissues via immunohistochemical staining. The biological functions of C4BPA were investigated in peripheral blood mononuclear cells (PBMCs) and human PDAC cell lines. Mouse C4BPA (mC4BPA) peptide, which is composed of 30 amino acids from the C-terminus and binds to CD40, was designed for further in vitro and in vivo experiments. In a preclinical experiment, we assessed the efficacy of gemcitabine plus nab-paclitaxel (GnP), dual immune checkpoint blockades (ICBs), and mC4BPA peptide in a mouse orthotopic transplantation model.
    RESULTS: Immunohistochemical analysis revealed that high stromal C4BPA and CD40 was associated with favorable PDAC prognosis (P=0.0005). Stromal C4BPA strongly correlated with the number of CD8+ tumor-infiltrating lymphocytes (P=0.001). In in vitro experiments, flow cytometry revealed that recombinant human C4BPA (rhC4BPA) stimulation increased CD4+ and CD8+ T cell numbers in PBMCs. rhC4BPA also promoted the proliferation of CD40-expressing PDAC cells. By contrast, combined treatment with gemcitabine and rhC4BPA increased PDAC cell apoptosis rate. mC4BPA peptide increased the number of murine T lymphocytes in vitro and the number of CD8+ tumor-infiltrating lymphocytes surrounding PDAC tumors in vivo. In a preclinical study, GnP/ICBs/mC4BPA peptide treatment, but not GnP treatment, led to the accumulation of a greater number of CD8+ T cells in the periphery of PDAC tumors and to greater tumor regression than did control treatment.
    CONCLUSIONS: These findings demonstrate that the combination of GnP therapy with C4BPA inhibits PDAC progression by promoting antitumor T cell accumulation in the tumor microenvironment.
    Keywords:  C4BPA; CD40; CD8; T cell antitumor immunogenicity; pancreatic cancer
    DOI:  https://doi.org/10.1186/s13046-021-02019-0
  7. Sci Rep. 2021 Jun 23. 11(1): 13144
    Impairing flow-mediated endothelial remodeling reduces extravasation of tumor cells.
    Gautier Follain, Naël Osmani, Valentin Gensbittel, Nandini Asokan, Annabel Larnicol, Luc Mercier, Maria Jesus Garcia-Leon, Ignacio Busnelli, Angelique Pichot, Nicodème Paul, Raphaël Carapito, Seiamak Bahram, Olivier Lefebvre, Jacky G Goetz.
      Tumor progression and metastatic dissemination are driven by cell-intrinsic and biomechanical cues that favor the growth of life-threatening secondary tumors. We recently identified pro-metastatic vascular regions with blood flow profiles that are permissive for the arrest of circulating tumor cells. We have further established that such flow profiles also control endothelial remodeling, which favors extravasation of arrested CTCs. Yet, how shear forces control endothelial remodeling is unknown. In the present work, we aimed at dissecting the cellular and molecular mechanisms driving blood flow-dependent endothelial remodeling. Transcriptomic analysis of endothelial cells revealed that blood flow enhanced VEGFR signaling, among others. Using a combination of in vitro microfluidics and intravital imaging in zebrafish embryos, we now demonstrate that the early flow-driven endothelial response can be prevented upon specific inhibition of VEGFR tyrosine kinase and subsequent signaling. Inhibitory targeting of VEGFRs reduced endothelial remodeling and subsequent metastatic extravasation. These results confirm the importance of VEGFR-dependent endothelial remodeling as a driving force of CTC extravasation and metastatic dissemination. Furthermore, the present work suggests that therapies targeting endothelial remodeling might be a relevant clinical strategy in order to impede metastatic progression.
    DOI:  https://doi.org/10.1038/s41598-021-92515-2
  8. Cell. 2021 Jun 17. pii: S0092-8674(21)00657-7. [Epub ahead of print]
    Expansion of tumor-associated Treg cells upon disruption of a CTLA-4-dependent feedback loop.
    Francesco Marangoni, Ademi Zhakyp, Michela Corsini, Shannon N Geels, Esteban Carrizosa, Martin Thelen, Vinidhra Mani, Jasper N Prüßmann, Ross D Warner, Aleksandra J Ozga, Mauro Di Pilato, Shivashankar Othy, Thorsten R Mempel.
      Foxp3+ T regulatory (Treg) cells promote immunological tumor tolerance, but how their immune-suppressive function is regulated in the tumor microenvironment (TME) remains unknown. Here, we used intravital microscopy to characterize the cellular interactions that provide tumor-infiltrating Treg cells with critical activation signals. We found that the polyclonal Treg cell repertoire is pre-enriched to recognize antigens presented by tumor-associated conventional dendritic cells (cDCs). Unstable cDC contacts sufficed to sustain Treg cell function, whereas T helper cells were activated during stable interactions. Contact instability resulted from CTLA-4-dependent downregulation of co-stimulatory B7-family proteins on cDCs, mediated by Treg cells themselves. CTLA-4-blockade triggered CD28-dependent Treg cell hyper-proliferation in the TME, and concomitant Treg cell inactivation was required to achieve tumor rejection. Therefore, Treg cells self-regulate through a CTLA-4- and CD28-dependent feedback loop that adjusts their population size to the amount of local co-stimulation. Its disruption through CTLA-4-blockade may off-set therapeutic benefits in cancer patients.
    Keywords:  CD28; CTLA-4; MP-IVM; NFAT; T regulatory cell; Treg cell; cytotoxic T lymphocyte-associated protein 4; multiphoton intravital microscopy; nuclear factor of activated T cells; tumor tolerance
    DOI:  https://doi.org/10.1016/j.cell.2021.05.027
  9. Nat Rev Immunol. 2021 Jun 21.
    Interferon-γ: teammate or opponent in the tumour microenvironment?
    Angela M Gocher, Creg J Workman, Dario A A Vignali.
      Cancer immunotherapy offers substantive benefit to patients with various tumour types, in some cases leading to complete tumour clearance. However, many patients do not respond to immunotherapy, galvanizing the field to define the mechanisms of pre-existing and acquired resistance. Interferon-γ (IFNγ) is a cytokine that has both protumour and antitumour activities, suggesting that it may serve as a nexus for responsiveness to immunotherapy. Many cancer immunotherapies and chemotherapies induce IFNγ production by various cell types, including activated T cells and natural killer cells. Patients resistant to these therapies commonly have molecular aberrations in the IFNγ signalling pathway or express resistance molecules driven by IFNγ. Given that all nucleated cells can respond to IFNγ, the functional consequences of IFNγ production need to be carefully dissected on a cell-by-cell basis. Here, we review the cells that produce IFNγ and the different effects of IFNγ in the tumour microenvironment, highlighting the pleiotropic nature of this multifunctional and abundant cytokine.
    DOI:  https://doi.org/10.1038/s41577-021-00566-3
  10. Cell Death Dis. 2021 Jun 25. 12(7): 649
    TNF-α induces endothelial-mesenchymal transition promoting stromal development of pancreatic adenocarcinoma.
    Marjorie Adjuto-Saccone, Philippe Soubeyran, Julie Garcia, Stéphane Audebert, Luc Camoin, Marion Rubis, Julie Roques, Bernard Binétruy, Juan Lucio Iovanna, Roselyne Tournaire.
      Endothelial-mesenchymal transition (EndMT) is an important source of cancer-associated fibroblasts (CAFs), which facilitates tumour progression. PDAC is characterised by abundant CAFs and tumour necrosis factor-α (TNF-α). Here, we show that TNF-α strongly induces human endothelial cells to undergo EndMT. Interestingly, TNF-α strongly downregulates the expression of the endothelial receptor TIE1, and reciprocally TIE1 overexpression partially prevents TNF-α-induced EndMT, suggesting that TNF-α acts, at least partially, through TIE1 regulation in this process. We also show that TNF-α-induced EndMT is reversible. Furthermore, TNF-α treatment of orthotopic mice resulted in an important increase in the stroma, including CAFs. Finally, secretome analysis identified TNFSF12, as a regulator that is also present in PDAC patients. With the aim of restoring normal angiogenesis and better access to drugs, our results support the development of therapies targeting CAFs or inducing the EndMT reversion process in PDAC.
    DOI:  https://doi.org/10.1038/s41419-021-03920-4
  11. Front Mol Biosci. 2021 ;8 645024
    Tumor Microenvironment Characteristics of Pancreatic Cancer to Determine Prognosis and Immune-Related Gene Signatures.
    Congjun Zhang, Jun Ding, Xiao Xu, Yangyang Liu, Wei Huang, Liangshan Da, Qiang Ma, Shengyang Chen.
      Background: Pancreatic cancer (PC) is one of the most lethal types of cancer with extremely poor diagnosis and prognosis, and the tumor microenvironment plays a pivotal role during PC progression. Poor prognosis is closely associated with the unsatisfactory results of currently available treatments, which are largely due to the unique pancreatic tumor microenvironment (TME). Methods: In this study, a total of 177 patients with PC from The Cancer Genome Atlas (TCGA) cohort and 65 patients with PC from the GSE62452 cohort in Gene Expression Omnibus (GEO) were included. Based on the proportions of 22 types of infiltrated immune cell subpopulations calculated by cell-type identification by estimating relative subsets of RNA transcripts (CIBERSORT), the TME was classified by K-means clustering and differentially expressed genes (DEGs) were determined. A combination of the elbow method and the gap statistic was used to explore the likely number of distinct clusters in the data. The ConsensusClusterPlus package was utilized to identify radiomics clusters, and the samples were divided into two subtypes. Result: Survival analysis showed that the patients with TMEscore-high phenotype had better prognosis. In addition, the TMEscore-high had better inhibitory effect on the immune checkpoint. A total of 10 miRNAs, 311 DEGs, and 68 methylation sites related to survival were obtained, which could be biomarkers to evaluate the prognosis of patients with pancreatic cancer. Conclusions: Therefore, a comprehensive description of TME characteristics of pancreatic cancer can help explain the response of pancreatic cancer to immunotherapy and provide a new strategy for cancer treatment.
    Keywords:  TMEscore; immune checkpoint; pancreatic cancer; prognosis; tumor microenvironment
    DOI:  https://doi.org/10.3389/fmolb.2021.645024
  12. Nat Commun. 2021 Jun 25. 12(1): 3969
    Molecular determinants of response to PD-L1 blockade across tumor types.
    Romain Banchereau, Ning Leng, Oliver Zill, Ethan Sokol, Gengbo Liu, Dean Pavlick, Sophia Maund, Li-Fen Liu, Edward Kadel, Nicole Baldwin, Suchit Jhunjhunwala, Dorothee Nickles, Zoe June Assaf, Daniel Bower, Namrata Patil, Mark McCleland, David Shames, Luciana Molinero, Mahrukh Huseni, Shomyseh Sanjabi, Craig Cummings, Ira Mellman, Sanjeev Mariathasan, Priti Hegde, Thomas Powles.
      Immune checkpoint inhibitors targeting the PD-1/PD-L1 axis lead to durable clinical responses in subsets of cancer patients across multiple indications, including non-small cell lung cancer (NSCLC), urothelial carcinoma (UC) and renal cell carcinoma (RCC). Herein, we complement PD-L1 immunohistochemistry (IHC) and tumor mutation burden (TMB) with RNA-seq in 366 patients to identify unifying and indication-specific molecular profiles that can predict response to checkpoint blockade across these tumor types. Multiple machine learning approaches failed to identify a baseline transcriptional signature highly predictive of response across these indications. Signatures described previously for immune checkpoint inhibitors also failed to validate. At the pathway level, significant heterogeneity is observed between indications, in particular within the PD-L1+ tumors. mUC and NSCLC are molecularly aligned, with cell cycle and DNA damage repair genes associated with response in PD-L1- tumors. At the gene level, the CDK4/6 inhibitor CDKN2A is identified as a significant transcriptional correlate of response, highlighting the association of non-immune pathways to the outcome of checkpoint blockade. This cross-indication analysis reveals molecular heterogeneity between mUC, NSCLC and RCC tumors, suggesting that indication-specific molecular approaches should be prioritized to formulate treatment strategies.
    DOI:  https://doi.org/10.1038/s41467-021-24112-w
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