bims-tuchim 2021-04-18 papers

bims-tuchim

Biomed News

on Tumor-on-chip models

Issue of 2021–04–18
ten papers selected by
Philipp Albrecht, Friedrich Schiller University

Studying Tumor Angiogenesis and Cancer Invasion in a Three-Dimensional Vascularized Breast Cancer Micro-Environment.
Functional heterogeneity of cytotoxic T cells and tumor resistance to cytotoxic hits limit anti-tumor activity in vivo.
Microvascularized tumor organoids-on-chips: advancing preclinical drug screening with pathophysiological relevance.
Engineered In Vitro Tumor Models for Cell-Based Immunotherapy.
Meet me halfway: Are in vitro 3D cancer models on the way to replace in vivo models for nanomedicine development?
Oncoimmunology Meets Organs-on-Chip.
An in vitro tumorigenesis model based on live-cell-generated oxygen and nutrient gradients.
Quantitative Evaluation of the Effect of Antigen Expression Level on Antibody-Drug Conjugate Exposure in Solid Tumor.
Stabilized epithelial phenotype of cancer cells in primary tumors leads to increased colonization of liver metastasis in pancreatic cancer.
Patient-Specific Organoid and Organ-on-a-Chip: 3D Cell-Culture Meets 3D Printing and Numerical Simulation.

Adv Biol (Weinh). 2021 Apr 15. e2100090

Studying Tumor Angiogenesis and Cancer Invasion in a Three-Dimensional Vascularized Breast Cancer Micro-Environment.

Madhuri Dey, Bugra Ayan, Marina Yurieva, Derya Unutmaz, Ibrahim T Ozbolat.

  Metastatic breast cancer is one of the deadliest forms of malignancy, primarily driven by its characteristic micro-environment comprising cancer cells interacting with stromal components. These interactions induce genetic and metabolic alterations creating a conducive environment for tumor growth. In this study, a physiologically relevant 3D vascularized breast cancer micro-environment is developed comprising of metastatic MDA-MB-231 cells and human umbilical vein endothelial cells loaded in human dermal fibroblasts laden fibrin, representing the tumor stroma. The matrix, as well as stromal cell density, impacts the transcriptional profile of genes involved in tumor angiogenesis and cancer invasion, which are hallmarks of cancer. Cancer-specific canonical pathways and activated upstream regulators are also identified by the differential gene expression signatures of these composite cultures. Additionally, a tumor-associated vascular bed of capillaries is established exhibiting dilated vessel diameters, representative of in vivo tumor physiology. Further, employing aspiration-assisted bioprinting, cancer-endothelial crosstalk, in the form of collective angiogenesis of tumor spheroids bioprinted at close proximity, is identified. Overall, this bottom-up approach of tumor micro-environment fabrication provides an insight into the potential of in vitro tumor models and enables the identification of novel therapeutic targets as a preclinical drug screening platform.

Keywords:  aspiration-assisted bioprinting; in vitro tumor models; metastatic breast cancer; tumor angiogenesis; tumor micro-environments

DOI:  https://doi.org/10.1002/adbi.202100090
EMBO J. 2021 Apr 15. e106658

Functional heterogeneity of cytotoxic T cells and tumor resistance to cytotoxic hits limit anti-tumor activity in vivo.

Roxana Khazen, Marine Cazaux, Fabrice Lemaître, Beatrice Corre, Zacarias Garcia, Philippe Bousso.

  Cytotoxic T cells (CTLs) can eliminate tumor cells through the delivery of lethal hits, but the actual efficiency of this process in the tumor microenvironment is unclear. Here, we visualized the capacity of single CTLs to attack tumor cells in vitro and in vivo using genetically encoded reporters that monitor cell damage and apoptosis. Using two distinct malignant B-cell lines, we found that the majority of cytotoxic hits delivered by CTLs in vitro were sublethal despite proper immunological synapse formation, and associated with reversible calcium elevation and membrane damage in the targets. Through intravital imaging in the bone marrow, we established that the majority of CTL interactions with lymphoma B cells were either unproductive or sublethal. Functional heterogeneity of CTLs contributed to diverse outcomes during CTL-tumor contacts in vivo. In the therapeutic settings of anti-CD19 CAR T cells, the majority of CAR T cell-tumor interactions were also not associated with lethal hit delivery. Thus, differences in CTL lytic potential together with tumor cell resistance to cytotoxic hits represent two important bottlenecks for anti-tumor responses in vivo.

Keywords:  CAR T cells; CTL; intravital imaging; lethal hit; sublethal hit

DOI:  https://doi.org/10.15252/embj.2020106658
Nano Converg. 2021 Apr 13. 8(1): 12

Microvascularized tumor organoids-on-chips: advancing preclinical drug screening with pathophysiological relevance.

Jungeun Lim, Hanna Ching, Jeong-Kee Yoon, Noo Li Jeon, YongTae Kim.

  Recent developments of organoids engineering and organ-on-a-chip microfluidic technologies have enabled the recapitulation of the major functions and architectures of microscale human tissue, including tumor pathophysiology. Nevertheless, there remain challenges in recapitulating the complexity and heterogeneity of tumor microenvironment. The integration of these engineering technologies suggests a potential strategy to overcome the limitations in reconstituting the perfusable microvascular system of large-scale tumors conserving their key functional features. Here, we review the recent progress of in vitro tumor-on-a-chip microfluidic technologies, focusing on the reconstruction of microvascularized organoid models to suggest a better platform for personalized cancer medicine.

Keywords:  Microfluidics; Microvasculature; Organ-on-a-chip; Tumor organoids

DOI:  https://doi.org/10.1186/s40580-021-00261-y
Acta Biomater. 2021 Apr 12. pii: S1742-7061(21)00234-8. [Epub ahead of print]

Engineered In Vitro Tumor Models for Cell-Based Immunotherapy.

Yuta Ando, Chelsea Mariano, Keyue Shen.

Tumor immunotherapy is rapidly evolving as one of the major pillars of cancer treatment. Cell-based immunotherapies, which utilize patient's own immune cells to eliminate cancer cells, have shown great promise in treating a range of malignancies, especially those of hematopoietic origins. However, their performance on a broader spectrum of solid tumor types still fall short of expectations in the clinical stage despite of the promising preclinical assessments. In this review, we briefly introduce cell-based immunotherapies and the inhibitory mechanisms in tumor microenvironments that may have contributed to this discrepancy. Specifically, a major obstacle to the clinical translation of cell-based immunotherapies is in the lack of preclinical models that can accurately assess the efficacies and mechanisms of these therapies in a (patho-)physiologically relevant manner. Lately, tissue engineering and organ-on-a-chip tools and microphysiological models have allowed for more faithful recapitulation of the tumor microenvironments, by incorporating crucial tumor tissue features such as cellular phenotypes, tissue architecture, extracellular matrix, physical parameters, and their dynamic interactions. This review summarizes the existing engineered tumor models with a focus on tumor immunology and cell-based immunotherapy. We also discuss some key considerations for the future development of engineered tumor models for immunotherapeutics. STATEMENT OF SIGNIFICANCE: Cell-based immunotherapies have shown great promise in treating hematological malignancies and some epithelial tumors. However, their performance on a broader spectrum of solid tumor types still fall short of expectations. Major obstacles include the inhibitory mechanisms in tumor microenvironments (TME) and the lack of preclinical models that can accurately assess the efficacies and mechanisms of cellular therapies in a (patho-)physiologically relevant manner. In this review, we introduce recent progress in tissue engineering and microphysiological models for more faithful recapitulation of TME for cell-based immunotherapies, and some key considerations for the future development of engineered tumor models. This overview will provide a better understanding on the role of engineered models in accelerating immunotherapeutic discoveries and clinical translations.

DOI: https://doi.org/10.1016/j.actbio.2021.03.076
Adv Drug Deliv Rev. 2021 Apr 07. pii: S0169-409X(21)00098-3. [Epub ahead of print]

Meet me halfway: Are in vitro 3D cancer models on the way to replace in vivo models for nanomedicine development?

Sabina Pozzi, Anna Scomparin, Sahar Israeli-Dangoor, Daniel Rodriguez, Paula Ofek, Lena Neufeld, Adva Krivitsky, Daniela Vaskovich, Ron Kleiner, Pradip Dey, Shani Koshrovski, Noa Reisman, Ronit Satchi-Fainaro.

  The complexity and diversity of the biochemical processes that occur during tumorigenesis and metastasis are frequently over-simplified in the traditional in vitro cell cultures. Two-dimensional cultures limit researchers' experimental observations and frequently give rise to misleading and contradictory results. Therefore, in order to overcome the limitations of in vitro studies and bridge the translational gap to in vivo applications, 3D models of cancer cell were developed in the last decades. The three dimensions of the tumor including its cellular and extracellular microenvironment are recreated by combining co-cultures of cancer and stromal cells in 3D hydrogel-based growth factors-inclusive scaffolds. More complex 3D cultures, containing functional blood vasculature, can integrate in the system external stimuli (e.g. oxygen and nutrient deprivation, cytokines, growth factors) along with drugs, or other therapeutic compounds. In this scenario, cell signaling pathways, metastatic cascade steps, cell differentiation and self-renewal, tumor-microenvironment interactions, precision and personalized medicine, are among the wide range of biological applications that can be studied using 3D tumor models. Here, we discuss a broad variety of strategies exploited by scientists to create in vitro 3D cancer models that resemble as much as possible the biology and patho-physiology of in vivo tumors and predict faithfully the treatment outcome.

Keywords:  3D models; drug screening; hydrogel-based ECM; immunotherapy; stromal cells interaction

DOI:  https://doi.org/10.1016/j.addr.2021.04.001
Front Mol Biosci. 2021 ;8 627454

Oncoimmunology Meets Organs-on-Chip.

Fabrizio Mattei, Sara Andreone, Arianna Mencattini, Adele De Ninno, Luca Businaro, Eugenio Martinelli, Giovanna Schiavoni.

  Oncoimmunology represents a biomedical research discipline coined to study the roles of immune system in cancer progression with the aim of discovering novel strategies to arm it against the malignancy. Infiltration of immune cells within the tumor microenvironment is an early event that results in the establishment of a dynamic cross-talk. Here, immune cells sense antigenic cues to mount a specific anti-tumor response while cancer cells emanate inhibitory signals to dampen it. Animals models have led to giant steps in this research context, and several tools to investigate the effect of immune infiltration in the tumor microenvironment are currently available. However, the use of animals represents a challenge due to ethical issues and long duration of experiments. Organs-on-chip are innovative tools not only to study how cells derived from different organs interact with each other, but also to investigate on the crosstalk between immune cells and different types of cancer cells. In this review, we describe the state-of-the-art of microfluidics and the impact of OOC in the field of oncoimmunology underlining the importance of this system in the advancements on the complexity of tumor microenvironment.

Keywords:  Cell on Chip; Oncoimmuno chip; Organ on Chip; cancer immunology; human on chip; microfluidic device; personalized medicine; tumor microenvironment

DOI:  https://doi.org/10.3389/fmolb.2021.627454
Commun Biol. 2021 Apr 15. 4(1): 477

An in vitro tumorigenesis model based on live-cell-generated oxygen and nutrient gradients.

Anne C Gilmore, Sarah J Flaherty, Veena Somasundaram, David A Scheiblin, Stephen J Lockett, David A Wink, William F Heinz.

The tumor microenvironment (TME) is multi-cellular, spatially heterogenous, and contains cell-generated gradients of soluble molecules. Current cell-based model systems lack this complexity or are difficult to interrogate microscopically. We present a 2D live-cell chamber that approximates the TME and demonstrate that breast cancer cells and macrophages generate hypoxic and nutrient gradients, self-organize, and have spatially varying phenotypes along the gradients, leading to new insights into tumorigenesis.

DOI: https://doi.org/10.1038/s42003-021-01954-0
AAPS J. 2021 Apr 15. 23(3): 56

Quantitative Evaluation of the Effect of Antigen Expression Level on Antibody-Drug Conjugate Exposure in Solid Tumor.

David Bussing, Sharad Sharma, Zhe Li, Lyndsey F Meyer, Dhaval K Shah.

  Antibody-drug conjugates (ADCs) rely on high expression of target antigens on cancer cells to effectively enter the cell and release a cytotoxic payload. Previous studies have shown that ADC efficacy is not always tied to antigen expression. However, our recent in vitro study suggests a linear relationship between antigen expression and the intracellular levels of the ADC payload. In this study, we have explored the relationship between antigen expression and intratumoral ADC exposure in vivo. Using trastuzumab-vc-MMAE (T-vc-MMAE) and four cell lines with varying expression of human epithelial growth factor receptor 2 (HER2), the pharmacokinetics of total trastuzumab, released ("free") MMAE, and total MMAE were evaluated in a tumor xenograft model. Nude mice were implanted with tumors originating from BT-474, MDA-MB-453, MCF-7, and MDA-MB-468 cell lines and dosed with 10 mg/kg or 1 mg/kg of ADC. Observed data were mathematically characterized using a mechanism-based PK model. A strong positive correlation was observed between antigen expression levels and free/total MMAE exposure (R2 ≥ 0.91) (total MMAE being the sum of released and conjugated MMAE) within the tumor, but not for total trastuzumab exposure. The PK model was able to recapitulate plasma PK through simulation; however, the tumor PK was overpredicted or underpredicted in some cases potentially due to differences in tumor vasculature or extracellular matrix conditions. Our results indicate a linear relationship between antigen expression and tumor exposure of free/total ADC payload in vivo, validating our previous finding in vitro, while also revealing the need to understand complex physiology of the tumor to predict tumor PK of ADC and its components. Our findings also support the concept of antigen expression screening in patients for targeted therapies like ADCs to achieve the maximum therapeutic benefit of the treatment.

Keywords:  antibody–drug conjugates; antigen expression level; solid tumor; systems PK model; tumor pharmacokinetics

DOI:  https://doi.org/10.1208/s12248-021-00584-y
Cell Rep. 2021 Apr 13. pii: S2211-1247(21)00304-1. [Epub ahead of print]35(2): 108990

Stabilized epithelial phenotype of cancer cells in primary tumors leads to increased colonization of liver metastasis in pancreatic cancer.

Julienne L Carstens, Sujuan Yang, Pedro Correa de Sampaio, Xiaofeng Zheng, Souptik Barua, Kathleen M McAndrews, Arvind Rao, Jared K Burks, Andrew D Rhim, Raghu Kalluri.

  Pancreatic ductal adenocarcinoma (PDAC) is therapeutically recalcitrant and metastatic. Partial epithelial to mesenchymal transition (EMT) is associated with metastasis; however, a causal connection needs further unraveling. Here, we use single-cell RNA sequencing and genetic mouse models to identify the functional roles of partial EMT and epithelial stabilization in PDAC growth and metastasis. A global EMT expression signature identifies ∼50 cancer cell clusters spanning the epithelial-mesenchymal continuum in both human and murine PDACs. The combined genetic suppression of Snail and Twist results in PDAC epithelial stabilization and increased liver metastasis. Genetic deletion of Zeb1 in PDAC cells also leads to liver metastasis associated with cancer cell epithelial stabilization. We demonstrate that epithelial stabilization leads to the enhanced collective migration of cancer cells and modulation of the immune microenvironment, which likely contribute to efficient liver colonization. Our study provides insights into the diverse mechanisms of metastasis in pancreatic cancer and potential therapeutic targets.

Keywords:  Snail; Twist; Zeb1; collective migration; epithelial-to-mesenchymal transition; immune modulation; metastasis; mouse models; pancreatic cancer; single-cell RNA sequencing

DOI:  https://doi.org/10.1016/j.celrep.2021.108990
Adv Biol (Weinh). 2021 Apr 15. e2000024

Patient-Specific Organoid and Organ-on-a-Chip: 3D Cell-Culture Meets 3D Printing and Numerical Simulation.

Fuyin Zheng, Yuminghao Xiao, Hui Liu, Yubo Fan, Ming Dao.

  The last few decades have witnessed diversified in vitro models to recapitulate the architecture and function of living organs or tissues and contribute immensely to advances in life science. Two novel 3D cell culture models: 1) Organoid, promoted mainly by the developments of stem cell biology and 2) Organ-on-a-chip, enhanced primarily due to microfluidic technology, have emerged as two promising approaches to advance the understanding of basic biological principles and clinical treatments. This review describes the comparable distinct differences between these two models and provides more insights into their complementarity and integration to recognize their merits and limitations for applicable fields. The convergence of the two approaches to produce multi-organoid-on-a-chip or human organoid-on-a-chip is emerging as a new approach for building 3D models with higher physiological relevance. Furthermore, rapid advancements in 3D printing and numerical simulations, which facilitate the design, manufacture, and results-translation of 3D cell culture models, can also serve as novel tools to promote the development and propagation of organoid and organ-on-a-chip systems. Current technological challenges and limitations, as well as expert recommendations and future solutions to address the promising combinations by incorporating organoids, organ-on-a-chip, 3D printing, and numerical simulation, are also summarized.

Keywords:  3D printing; numerical simulations; organ-on-a-chip; organoids; synergistic engineering

DOI:  https://doi.org/10.1002/adbi.202000024