Organ Model: Liver

Organ Model: Liver

Application: Immunology & Inflammation

Abstract: Immunotherapy has changed the landscape of treatment options for patients with hepatocellular cancer. Checkpoint inhibitors are now standard of care for patients with advanced tumours, yet the majority remain resistant to this therapy and urgent approaches are needed to boost the efficacy of these agents. Targeting the liver endothelial cells, as the orchestrators of immune cell recruitment, within the tumour microenvironment of this highly vascular cancer could potentially boost immune cell infiltration. We demonstrate the successful culture of primary human liver endothelial cells in organ-on-a-chip technology followed by perfusion of peripheral blood mononuclear cells. We confirm, with confocal and multiphoton imaging, the capture and adhesion of immune cells in response to pro-inflammatory cytokines in this model. This multicellular platform sets the foundation for testing the efficacy of new therapies in promoting leukocyte infiltration across liver endothelium as well as a model for testing cell therapy, such as chimeric antigen receptor (CAR)-T cell, capture and migration across human liver endothelium.

Overview

Drug induced Liver-Injury (DILI) is a leading cause of drug attrition, and complex in vitro models (CIVMs)—including three dimensional (3D) spheroids, Organ-Chips, 3D bio printed tissues, and flow-based systems—could improve preclinical prediction. Although CIVMs have demonstrated good sensitivity and specificity, in DILI detection their adoption remains limited.

This article describes DILI, the challenges with its prediction and the current strategies, and the models that are being used to study it. It reviews data from industry-FDA collaborations and strategic partnerships and finishes with an outlook of CIVMs in preclinical toxicity testing.

Abstract

Bile acids are an essential component of bile, which aids digestion in the form of emulsification and absorption of lipids. Primary bile acids (cholic and chenodeoxycholic acid) are synthesized in the liver, can be conjugated with glycine or taurine, concentrated in the gall bladder, and circulated to the intestine during digestion. Primary bile acids are converted to secondary bile acids (deoxycholic and lithocholic acid) by intestinal bacteria and are recirculated back to the liver via enterohepatic circulation. In a healthy liver, bile acids are secreted from hepatocytes into bile canaliculi through the bile salt export pump (BSEP) which is involved in maintaining bile acid homeostasis. When bile flow is disrupted, increasing levels of intrahepatic bile acids cause cholestatic injury. Cholestasis can be induced by drugs like troglitazone (TROG), a diabetes therapy that was discontinued after causing liver toxicity in clinical trials partly via inhibition of BSEP. To establish a physiologically relevant model of drug-induced cholestasis, Emulate’s human Quad-Culture Liver-Chip was incubated with bile acids in the presence or absence of TROG and monitored for signs of toxicity and BSEP inhibition. A mixture of glycine-conjugated primary and secondary bile acids, including glycocholic acid (GCA), glycodeoxycholic acid (GDCA) and glycochenodeoxycholic acid (GCDCA), were chosen as they are three of the most abundant bile acids in human serum (comprising ~68% of the
total population) and were administered at concentrations ranging from 1-5X human Cmax.

Abstract

The field of cancer immunotherapy is rapidly growing but is challenged by our inability to predict treatment efficacy, patient response and adverse effects. Agonistic antibodies toward T cell co-stimulatory molecules like Urelumab (targeting CD137) are one class of immunotherapy that has shown unparalleled efficacy in murine models of cancer. However, in clinical trials (NCT00309023), Urelumab doses ≥ 1 mg/kg resulted in significant hepatotoxicity (mainly transaminitis), suggesting murine and NHP preclinical assessment failed to predict this clinical safety outcome. While the mechanism of clinical liver toxicity has not been fully elucidated, studies with humanized mice (with human hematopoietic cells) indicate immune-mediated DILI (drug-induced liver injury). Therefore, to model hepatotoxicity associated with Urelumab, we created a novel, immuno-competent human Quad-Culture Liver-Chip using human peripheral blood mononuclear cells (PBMCs) that contain lymphocytes, NK cells and CD137+ monocytes to drive immunotoxicity.

Organ Model: Liver, Intestine (Caco2)

Application: Organ transplantation

How Organ-Chips Were Used: Drugs that induce reversible slowing of metabolic and physiological processes would have great value for organ preservation, especially for organs with high susceptibility to hypoxia-reperfusion injury, such as the heart. Here, Gut- and Liver-Chips were used to evaluate the metabolic suppression capability of the test compound. The Chips showed a decrease in the tissue’s total ATP production in presence of SNC80, which is accompanied by a global slowing of metabolism.

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Webinar Abstract

Featured session at Boston MPS Day, which took place on 11/1/2023.

In this presentation, Dr. Samantha Atkins from Moderna discusses the use of the Emulate human Liver-Chip to predict and understand toxicity mechanisms associated with lipid nanoparticle (LNP) and mRNA therapeutics. Traditional animal models, such as rodent studies, sometimes fail to predict adverse outcomes seen later in non-human primate (NHP) studies or clinical settings, particularly when novel LNP chemistries are involved. By leveraging the Emulate human Liver-Chip, Dr. Atkins aims to identify early signatures of fibrosis and other forms of liver damage, thereby guiding safer and more effective therapeutic design before proceeding to NHP studies.

Using the Liver-Chips, Dr. Atkins’ team demonstrated the ability to pinpoint gene signatures and collagen remodeling patterns corresponding to pro-fibrotic LNPs. This helps flag problematic compounds early, reducing reliance on costly and time-consuming animal studies. Moreover, the chips provided mechanistic insights into both LNP-driven and mRNA-mediated toxicity, revealing pathways such as necroptosis in certain constructs. Such mechanistic understanding helps tailor safer LNP formulations and mRNA constructs.

Key learnings from this presentation include:

• Human liver-on-a-chip systems can successfully model liver fibrosis and flag toxicity that might be missed by animal models.
• Early gene signatures identified on the chip correlate with adverse outcomes in NHP models, enabling preclinical down-selection of safer LNP candidates.
• The platform aids in distinguishing between LNP-induced versus mRNA-mediated toxicity, offering valuable mechanistic insights.
• Leveraging these in vitro models can save time and reduce costs associated with late-stage animal testing.
• Mechanistic understanding of cell death pathways (e.g., necroptosis) informs rational modifications to improve therapeutic safety profiles.

Featured session at Bethesda MPS Day, which took place on November 9, 2023.

Dr. Dylan Fudge from the U.S. Army DEVCOM Chemical Biological Center discusses the use of Organ-Chip models, specifically Emulate’s MPS platforms, to advance the Army’s predictive toxicology efforts related to chemical and biological threats. Traditional safety and efficacy assessments often rely heavily on animal models and simplistic in vitro assays, which may not accurately reflect human physiology, particularly for acute, high-risk agents. By integrating human-relevant cells into microfluidic devices, Dr. Fudge’s team aims to improve mechanistic understanding, dose-response assessments, and long-term impact evaluations of nerve agents, PFAS compounds, and pathogens like SARS-CoV-2.

He showcases three major projects:

1. VX Nerve Agent on Liver-Chip:
   Exposing a human Liver-Chip (with hepatocytes, endothelial cells, stellates, and Kupffer cells) to sub-lethal VX concentrations allowed for multi-omics analyses (proteomics, metabolomics, and transcriptomics). The system revealed shifts in metabolic pathways, bioenergetics, and nitrogen metabolism consistent with in vivo and clinical literature. Importantly, VX’s known mechanism—disruption of acetylcholine breakdown—was mirrored in altered choline metabolism, reinforcing the Liver-Chip’s translational relevance.
2. PFAS Exposure in Kidney and Liver-Chips:
   Investigating polyfluoroalkyl substances (PFAS), the team saw patterns of compound retention and toxicity aligned with known clinical effects. Liver and Kidney-Chips demonstrated differential absorption and metabolic responses to PFAS, supported by robust omics data. Oxygen stress markers and biomarkers like uric acid levels recapitulated known toxicity profiles, indicating the chips’ capacity to model chronic, low-level exposures and related subtler health effects.
3. SARS-CoV-2 Infection Using Lung-Chips:
   Lung-Chips incorporating airway and alveolar regions enabled controlled infection with SARS-CoV-2 variants. Despite minimal acute cytotoxicity, omics analyses detected hallmark inflammatory pathways and mechanistic insights matching emerging clinical findings—such as immune activation and potential neuro-related disease pathways—emphasizing the chip’s ability to reveal complex host-pathogen interactions.

Collectively, these studies highlight the versatility and reliability of Emulate’s Organ-Chip platforms under rigorous defense-oriented research conditions. Their capacity to maintain viable tissues, capture human-like biology, and provide high-content data (via omics) positions these systems as valuable tools to reduce animal use, support therapeutic development, and enhance understanding of chemical and biological threat agents at a mechanistic level.

Drug-induced liver injury (DILI) is a potentially lethal condition that heavily impacts the pharmaceutical industry, causing approximately 21% of drug withdrawals and 13% of clinical trial failures. Recent evidence suggests that the use of Liver-Chip technology in preclinical safety testing may significantly reduce DILI-related clinical trial failures and withdrawals. However, drug developers and regulators would benefit from guidance on the integration of Liver-Chip data into decision-making processes to facilitate the technology’s adoption.

This perspective builds on the findings of the performance assessment of the Emulate Liver-Chip in the context of DILI prediction and introduces two new decision-support frameworks: the first uses the Liver-Chip’s quantitative output to elucidate DILI severity and enable more nuanced risk analysis; the second integrates Liver-Chip data with standard animal testing results to help assess whether to progress a candidate drug into clinical trials.

There is now strong evidence that Liver-Chip technology could significantly reduce the incidence of DILI in drug development. As this is a patient safety issue, it is imperative that developers and regulators explore the incorporation of the technology. The frameworks presented enable the integration of the Liver-Chip into various stages of preclinical development in support of safety assessment.

Webinar Abstract

Adeno-associated virus (AAV)-derived vectors have emerged as a promising gene delivery vehicle for a broad range of clinical indications. The liver in particular is an interesting target as it is easily transduced with AAV vectors and allows a persistent expression of the transgene. However, off-target gene transduction remains a significant challenge in the development of successful gene therapeutics; thus, it is crucial to develop efficient organ-specific promoters for gene therapy constructs. While simple in vitro 2D cell culture models are useful for initial higher throughput screening, their limited complexity often results in responses that do not translate to animal models or human patients. Organ-on-a-Chip technology is helping to bridge this gap by enabling researchers to assess gene therapy constructs in a more physiologically relevant microenvironment. 

In this webinar, Dr. Rui Sun, Research Scientist at Bayer AG, presented on how he applied the Emulate Liver-Chip to evaluate the transduction efficiency and selectivity of liver-specific AAV promoters.

Learn how the Liver-Chip can be used to:

• Improve the evaluation of AAV vector transduction efficiency 
• Rank order various candidate promoter constructs 
• Quantify transgene expression efficiency and specificity 
• Investigate transduction efficiency by cell type
• Evaluate promoter efficacy in a liver disease model

Webinar Abstract

Drug-induced liver injury (DILI) is a potentially lethal condition that heavily impacts the pharmaceutical industry, causing drug withdrawals and approximately 13% of clinical trial failures. The prevalence of DILI in clinical settings exposes the limitations of preclinical toxicology models, which fail to capture the complexity of human liver physiology and its response to drug exposure.  

Recent evidence suggests that adopting the Emulate Liver-Chip into preclinical workflows may reduce DILI-related clinical trial failures. Here, we present a quantitative framework for integrating Liver-Chip data into the pharmaceutical decision-making process. This new Liver-Chip DILI score enables researchers to categorize tested compounds into standard DILI risk categories utilizing human Liver-Chip and animal in vivo data.  

During this webinar, topics that were covered included: 

• Characterization results of the Emulate Liver-Chip within the context of use of DILI prediction 
• Where in the preclinical workflow the Liver-Chip can be deployed 
• How the Liver-Chip’s quantitative readouts can be used to map prospective therapeutics into standard DILI risk categories  
• Methodology for combining Liver-Chip data with animal in vivo data, allowing for a more holistic assessment of DILI risk ahead of clinical trial testing
  

The presentation was followed by an informative Q&A session.