Search

Human organ-on-a-chip (Organ-Chip) technology has the potential to disrupt preclinical drug discovery and improve success in drug development pipelines as it can recapitulate organ-level pathophysiology and clinical responses. The Innovation and Quality (IQ) consortium formed by multiple pharmaceutical and biotechnology companies to confront this challenge has published guidelines that define criteria for qualifying preclinical models, however, systematic and quantitative evaluation of the predictive value of Organ-Chips has not yet been reported. Here, 870 Liver-Chips were analyzed to determine their ability to predict drug-induced liver injury (DILI) caused by small molecules identified as benchmarks by the IQ consortium. The Liver-Chip met the qualification guidelines across a blinded set of 27 known hepatotoxic and non-toxic drugs with a sensitivity of 87% and a specificity of 100%. A computational economic value analysis suggests that with this performance the Liver-Chip could generate $3 billion annually for the pharmaceutical industry due to increased R&D productivity.

The intestinal epithelial barrier supports the symbiotic relationship between the microbiota colonizing the intestinal epithelium and the host immune system to maintain homeostasis. Leaky barrier is increasingly recognized as part of the pathogenesis of a number of chronic conditions in addition to inflammatory and infectious diseases. As our understanding on the regulation of the barrier remains limited, effective therapeutic targeting for the compromised barrier is still an unmet need. Here we combined advancements on the organoids and Organ-on-Chip technologies to establish a micro-engineered Colon Intestine-Chip for studying development and regulation of the human intestinal barrier. Our data demonstrate the significance of the endothelium in co-culture with the epithelial cells within a tissue-relevant microenvironment for the establishment of a tight epithelial barrier of polarized cells. Pathway analysis of the RNA sequencing (RNA-Seq), revealed significant upregulation of mechanisms relevant to the maturation of the intestinal epithelium in organoid-derived epithelial cells in co-culture with endothelium as compared to organoids maintained in suspension. We provide evidence that the Colon Intestine-Chip platform responds to interferon gamma (IFNγ), a prototype cytokine utilized to model inflammation-induced barrier disruption, by induction of apoptosis and reorganization of the apical junctional complexes as shown with other systems. We also describe the mechanism of action of interleukin 22 (IL-22) on mature, organoid-derived intestinal epithelial cells that is consistent with barrier disruption. Overall we propose the Colon Intestine-Chip as a promising human organoid-derived platform to decipher mechanisms driving the development of leaky gut in patients and enable their translation for this unmet medical need.

Successful translation ofin vivoexperimental data to human patients is an unmet need and a bottleneck in the development of effective therapeutics. micro technology aims to address this need with significant advancements reported recently that enable modeling of organ level function. These microengineered chips enable researcher to recreate critical elements such asin vivorelevant tissue-tissue interface, air-liquid interface, and mechanical forces, such as mechanical stretch and fluidic shear stress, are crucial in emulating tissue level functions. Here, we present the development of a new, comprehensive 3D cell-culture system, where we combined our proprietary Organ-Chip technology with recent advantages in three-dimensional organotypic culture. Leveraging microfabrication techniques, we engineered a flexible chip that consists of a channel containing an organotypic epithelium surrounded by two vacuum channels that can be actuated to stretch the hydrogel throughout its thickness. Furthermore, the ceiling of this channel is a removable lid with a built-in microchannel that can be perfused with liquid or air and removed as needed for direct access to the tissue. The floor of this channel is a porous flexible membrane in contact with a microfluidic channel that provides diffusive mass transport to and from the channel. This additional microfluidic channel can be coated with endothelial cells to emulate a blood vessel and capture endothelial interactions. Our results show that the Open-Top Chip design successfully addresses common challenges associated with the Organs-on-Chips technology, including the capability to incorporate a tissue-specific extracellular matrix gel seeded with primary stromal cells, to reproduce the architectural complexity of tissues by micropatterning the gel, that can be extracted for H&E staining. We provide proof-of-concept data on the feasibility of the system using skin and alveolar epithelial primary cells and by simulating alveolar inflammation.

RationaleViral-induced exacerbation of asthma remain a major cause of hospitalization and mortality. New human relevant models of the airways are urgently needed to understand how respiratory infections may trigger asthma attacks, and to advance treatment development.ObjectivesTo develop a new human relevant model of rhinovirus-induced asthma exacerbation that recapitulates viral infection of asthmatic airway epithelium, neutrophil transepithelial migration, and enables evaluation of immunomodulatory therapy.MethodsA micro-engineered model of fully differentiated human mucociliary airway epithelium was stimulated with IL-13 to induce a Th2-type asthmatic phenotype and infected with live human rhinovirus 16 (HRV16) to reproduce key features of viral-induced asthma exacerbation.Measurements and Main ResultsInfection with HRV16 replicated key hallmarks of the cytopathology and inflammatory responses observed in human airways. Generation of a Th2 microenvironment through exogenous IL-13 stimulation induced features of asthmatics airways, including goblet cell hyperplasia, reduction of cilia beating frequency, and endothelial activation, but did not alter rhinovirus infectivity or replication. High resolution kinetic analysis of secreted inflammatory markers revealed that IL-13 treatment altered the IL-6, IFN-λ1, and CXCL10 secretion in response to HRV16. Neutrophil transepithelial migration was greatest when viral infection was combined with IL-13 treatment, while treatment with MK-7123, a CXCR2 antagonist, reduced neutrophil diapedesis in all conditions.ConclusionsThis micro-engineered Airway Lung-Chip provides a novel human-relevant platform for exploring the complex mechanisms underlying viral-induced asthma exacerbation. Our data suggest that IL-13 may impair the hosts’ ability to mount an appropriate and coordinated immune response to rhinovirus infection. We also show that the Airway Lung-Chip can be used to assess the efficacy of modulators of the immune response.NoteEmulate®, Human Emulation System®, S-1™, ER-1™, and ER-2™ are trademarks of Emulate, Inc., and any other trademarks used herein remain with their respective holders. The technology disclosed in this document may be covered by one or more patents or patent applications, and no license to these is granted herein. You are solely responsible for determining whether you have all intellectual property rights that are necessary for your intended use of any of the disclosed materials, and whether you are required to obtain any additional intellectual property rights from a third party. Further information is available by contacting the authors.At a Glance CommentaryScientific Knowledge on the SubjectNew therapies for asthma exacerbations remain a significant unmet medical need. Development of human relevant preclinical models are needed to further elucidate the complex mechanisms underlying asthma exacerbation and investigate new therapeutic strategies.What This Study Adds to the FieldUsing a human Airway Lung-Chip model, we show here for the first time a live human rhinovirus (HRV) infection of the asthmatic epithelium that recapitulates complex features of viral-induced asthma exacerbation. The dynamic microenvironment of the chip enables the real-time study of virus infection, epithelial response, and immune cell recruitment under healthy and asthmatic conditions. The model reproduces key endpoints that have been observed in asthmatics and individuals infected with rhinovirus including the ciliated cell sloughing, altered cilia beating frequency, goblet cell hyperplasia, increased expression of adhesion molecules in microvascular endothelial cells, and inflammatory mediator release. High-resolution temporal analysis of secreted inflammatory markers enabled by dynamic sampling revealed alteration of IL-6, IFN-λ1 and CXCL10 secretory phases after rhinovirus infection in an IL-13 high environment. Leveraging high-content imaging and analysis of circulating inflammatory cells, we demonstrated the efficacy of a CXCR2 antagonist to reduce adhesion, motility, and transmigration of perfused human neutrophils. Thus, this micro-engineered chip may offer a powerful addition to preclinical models for understanding mechanisms underlying asthma exacerbation pathology and developing new therapeutic strategies.