bims-tuchim Biomed News
on Tumor-on-chip models
Issue of 2021‒08‒01
six papers selected by
Philipp Albrecht
Friedrich Schiller University
  1. Investigating lymphangiogenesis in vitro and in vivo using engineered human lymphatic vessel networks.
  2. Cancer-Associated Fibroblast Heterogeneity: A Factor That Cannot Be Ignored in Immune Microenvironment Remodeling.
  3. Mammary Epithelial and Endothelial Cell Spheroids as a Potential Functional In vitro Model for Breast Cancer Research.
  4. Cell death in pancreatic cancer: from pathogenesis to therapy.
  5. 3D Bioprinted Multicellular Vascular Models.
  6. A robust vasculogenic microfluidic model using human immortalized endothelial cells and Thy1 positive fibroblasts.
  1. Proc Natl Acad Sci U S A. 2021 Aug 03. pii: e2101931118. [Epub ahead of print]118(31):
    Investigating lymphangiogenesis in vitro and in vivo using engineered human lymphatic vessel networks.
    Shira Landau, Abigail Newman, Shlomit Edri, Inbal Michael, Shahar Ben-Shaul, Yulia Shandalov, Tom Ben-Arye, Pritinder Kaur, Ming H Zheng, Shulamit Levenberg.
      The lymphatic system is involved in various biological processes, including fluid transport from the interstitium into the venous circulation, lipid absorption, and immune cell trafficking. Despite its critical role in homeostasis, lymphangiogenesis (lymphatic vessel formation) is less widely studied than its counterpart, angiogenesis (blood vessel formation). Although the incorporation of lymphatic vasculature in engineered tissues or organoids would enable more precise mimicry of native tissue, few studies have focused on creating engineered tissues containing lymphatic vessels. Here, we populated thick collagen sheets with human lymphatic endothelial cells, combined with supporting cells and blood endothelial cells, and examined lymphangiogenesis within the resulting constructs. Our model required just a few days to develop a functional lymphatic vessel network, in contrast to other reported models requiring several weeks. Coculture of lymphatic endothelial cells with the appropriate supporting cells and intact PDGFR-β signaling proved essential for the lymphangiogenesis process. Additionally, subjecting the constructs to cyclic stretch enabled the creation of complex muscle tissue aligned with the lymphatic and blood vessel networks, more precisely biomimicking native tissue. Interestingly, the response of developing lymphatic vessels to tensile forces was different from that of blood vessels; while blood vessels oriented perpendicularly to the stretch direction, lymphatic vessels mostly oriented in parallel to the stretch direction. Implantation of the engineered lymphatic constructs into a mouse abdominal wall muscle resulted in anastomosis between host and implant lymphatic vasculatures, demonstrating the engineered construct's potential functionality in vivo. Overall, this model provides a potential platform for investigating lymphangiogenesis and lymphatic disease mechanisms.
    Keywords:  engineered tissue; lymphangiogenesis; lymphatic endothelial cells; vascularization
    DOI:  https://doi.org/10.1073/pnas.2101931118
  2. Front Immunol. 2021 ;12 671595
    Cancer-Associated Fibroblast Heterogeneity: A Factor That Cannot Be Ignored in Immune Microenvironment Remodeling.
    Pei-Yu Chen, Wen-Fei Wei, Hong-Zhen Wu, Liang-Sheng Fan, Wei Wang.
      Cancer-associated fibroblasts (CAFs) are important, highly heterogeneous components of the tumor extracellular matrix that have different origins and express a diverse set of biomarkers. Different subtypes of CAFs participate in the immune regulation of the tumor microenvironment (TME). In addition to their role in supporting stromal cells, CAFs have multiple immunosuppressive functions, via membrane and secretory patterns, against anti-tumor immunity. The inhibition of CAFs function and anti-TME therapy targeting CAFs provides new adjuvant means for immunotherapy. In this review, we outline the emerging understanding of CAFs with a particular emphasis on their origin and heterogeneity, different mechanisms of their regulation, as well as their direct or indirect effect on immune cells that leads to immunosuppression.
    Keywords:  cancer-associated fibroblasts; heterogeneity; immune evasion; tumor immunology; tumor microenvironment
    DOI:  https://doi.org/10.3389/fimmu.2021.671595
  3. J Vis Exp. 2021 Jul 12.
    Mammary Epithelial and Endothelial Cell Spheroids as a Potential Functional In vitro Model for Breast Cancer Research.
    Giovanna Azzarito, Marta Ewa Szutkowska, Annalisa Saltari, Edwin K Jackson, Brigitte Leeners, Marinella Rosselli, Raghvendra K Dubey.
      Breast cancer is the leading cause of mortality in women. The growth of breast cancer cells and their subsequent metastasis is a key factor for its progression. Although the mechanisms involved in promoting breast cancer growth have been intensively studied using monocultures of breast cancer cells such as MCF-7 cells, the contribution of other cell types, such as vascular and lymphatic endothelial cells that are intimately involved in tumor growth, has not been investigated in depth. Cell-cell interaction plays a key role in tumor growth and progression. Neoangiogenesis, or the development of vessels, is essential for tumor growth, whereas the lymphatic system serves as a portal for cancer cell migration and subsequent metastasis. Recent studies provide evidence that vascular and lymphatic endothelial cells can significantly influence cancer cell growth. These observations imply a need for developing in vitro models that would more realistically reflect breast cancer growth processes in vivo. Moreover, restrictions in animal research require the development of ex vivo models to elucidate better the mechanisms involved. This article describes the development of breast cancer spheroids composed of both breast cancer cells (estrogen receptor-positive MCF-7 cells) and vascular and/or lymphatic endothelial cells. The protocol describes a detailed step-by-step approach in creating dual-cell spheroids using two different approaches, hanging drop (gold standard and cheap) and 96-well U-bottom plates (expensive). In-depth instructions are provided for how to delicately pick up the formed spheroids to monitor growth by microscopic sizing and assessing viability using dead and live cell staining. Moreover, procedures to fix the spheroids for sectioning and staining with growth-specific antibodies to differentiate growth patterns in spheroids are delineated. Additionally, details for preparing spheroids with transfected cells and methods to extract RNA for molecular analysis are provided. In conclusion, this article provides in-depth instructions for preparing multi-cell spheroids for breast cancer research.
    DOI:  https://doi.org/10.3791/62940
  4. Nat Rev Gastroenterol Hepatol. 2021 Jul 30.
    Cell death in pancreatic cancer: from pathogenesis to therapy.
    Xin Chen, Herbert J Zeh, Rui Kang, Guido Kroemer, Daolin Tang.
      Pancreatic cancer is a devastating gastrointestinal cancer characterized by late diagnosis, limited treatment success and dismal prognosis. Exocrine tumours account for 95% of pancreatic cancers and the most common pathological type is pancreatic ductal adenocarcinoma (PDAC). The occurrence and progression of PDAC involve multiple factors, including internal genetic alterations and external inflammatory stimuli. The biology and therapeutic response of PDAC are further shaped by various forms of regulated cell death, such as apoptosis, necroptosis, ferroptosis, pyroptosis and alkaliptosis. Cell death induced by local or systemic treatments suppresses tumour proliferation, invasion and metastasis. However, unrestricted cell death or tissue damage might result in an inflammation-related immunosuppressive microenvironment, which is conducive to tumour progression or recurrence. The precise extent to which cell death affects PDAC is not yet well described. A growing body of preclinical and clinical studies document significant correlations between mutations (for example, in KRAS and TP53), stress responses (such as hypoxia and autophagy), metabolic reprogramming and chemotherapeutic responses. Here, we describe the molecular machinery of cell death, discuss the complexity and multifaceted nature of lethal signalling in PDAC cells, and highlight the challenges and opportunities for activating cell death pathways through precision oncology treatments.
    DOI:  https://doi.org/10.1038/s41575-021-00486-6
  5. Adv Healthc Mater. 2021 Jul 26. e2101141
    3D Bioprinted Multicellular Vascular Models.
    Karli A Gold, Biswajit Saha, Navaneeth Krishna Rajeeva Pandian, Brandon K Walther, Jorge A Palma, Javier Jo, John P Cooke, Abhishek Jain, Akhilesh K Gaharwar.
      3D bioprinting is an emerging additive manufacturing technique to fabricate constructs for human disease modeling. However, current cell-laden bioinks lack sufficient biocompatibility, printability, and structural stability needed to translate this technology to preclinical and clinical trials. Here, a new class of nanoengineered hydrogel-based cell-laden bioinks is introduced, that can be printed into 3D, anatomically accurate, multicellular blood vessels to recapitulate both the physical and chemical microenvironments of native human vasculature. A remarkably unique characteristic of this bioink is that regardless of cell density, it demonstrates a high printability and ability to protect encapsulated cells against high shear forces in the bioprinting process. 3D bioprinted cells maintain a healthy phenotype and remain viable for nearly one-month post-fabrication. Leveraging these properties, the nanoengineered bioink is printed into 3D cylindrical blood vessels, consisting of living co-culture of endothelial cells and vascular smooth muscle cells, providing the opportunity to model vascular function and pathophysiology. Upon cytokine stimulation and blood perfusion, this 3D bioprinted vessel is able to recapitulate thromboinflammatory responses observed only in advanced in vitro preclinical models or in vivo. Therefore, this 3D bioprinted vessel provides a potential tool to understand vascular disease pathophysiology and assess therapeutics, toxins, or other chemicals.
    Keywords:  3D bioprinting; cell-laden bioink; disease models; regenerative medicine; vascular tissue
    DOI:  https://doi.org/10.1002/adhm.202101141
  6. Biomaterials. 2021 Jul 16. pii: S0142-9612(21)00388-4. [Epub ahead of print]276 121032
    A robust vasculogenic microfluidic model using human immortalized endothelial cells and Thy1 positive fibroblasts.
    Zhengpeng Wan, Shun Zhang, Amy X Zhong, Sarah E Shelton, Marco Campisi, Shriram K Sundararaman, Giovanni S Offeddu, Eunkyung Ko, Lina Ibrahim, Mark F Coughlin, Tiankun Liu, Jing Bai, David A Barbie, Roger D Kamm.
      Human umbilical vein endothelial cells (HUVECs) and stromal cells, such as human lung fibroblasts (FBs), have been widely used to generate functional microvascular networks (μVNs) in vitro. However, primary cells derived from different donors have batch-to-batch variations and limited lifespans when cultured in vitro, which hampers the reproducibility of μVN formation. Here, we immortalize HUVECs and FBs by exogenously expressing human telomerase reverse transcriptase (hTERT) to obtain stable endothelial cell and FB sources for μVN formation in vitro. Interestingly, we find that immortalized HUVECs can only form functional μVNs with immortalized FBs from earlier passages but not from later passages. Mechanistically, we show that Thy1 expression decreases in FBs from later passages. Compared to Thy1 negative FBs, Thy1 positive FBs express higher IGFBP2, IGFBP7, and SPARC, which are important for angiogenesis and lumen formation during vasculogenesis in 3D. Moreover, Thy1 negative FBs physically block microvessel openings, reducing the perfusability of μVNs. Finally, by culturing immortalized FBs on gelatin-coated surfaces in serum-free medium, we are able to maintain the majority of Thy1 positive immortalized FBs to support perfusable μVN formation. Overall, we establish stable cell sources for μVN formation and characterize the functions of Thy1 positive and negative FBs in vasculogenesis in vitro.
    Keywords:  Fibroblasts; HUVECs; Immortalization; Microfluidic; Thy1; Vasculogenesis
    DOI:  https://doi.org/10.1016/j.biomaterials.2021.121032
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