Organ Model: Liver

Webinar Abstract

In December 2022, two milestone papers were published: “Predictive validity in drug discovery: what it is, why it matters and how to improve it” in NRDD and “Performance assessment and economic analysis of a human Liver-Chip for predictive toxicology” in Communications Medicine, part of Nature Portfolio. In addition, President Biden signed the FDA Modernization Act 2.0 into law. This groundbreaking advancement allows researchers to leverage Organ-on-a-Chip technology data as part of a novel IND submission, breaking the 1938 Food, Drug, and Cosmetic Act (FDCA) mandate that required all new drugs be tested on animals.

To say the least, it was an exciting way to close out the year. However, these milestones are sparking many questions across the industry: What’s next, how does this impact my work, and how do I get started with Organ-on-a-Chip technology?

Watch this on-demand fireside chat to see industry experts discuss:

• What is predictive validity in drug discovery, why is it important, and how can we improve it?
• The recent Communications Medicine publication demonstrated that the Emulate human Liver-Chip was able to correctly identify 87% of the tested drugs that caused drug-induced liver injury in patients despite passing animal testing evaluations—and did not falsely identify any drugs as toxic, leading to a 100% specificity rating. We will discuss how this will help your decision-making criteria to improve patient safety and increase your confidence that a viable candidate drug will not be de-prioritized due to a false positive.
• How could an improvement in sensitivity and specificity lead to a $3B increase in R&D productivity for small molecule drug development?
• And lastly, what can we expect with the passing of the FDA Modernization Act and the growing global movement to eliminate animal testing mandates?

Abstract

As cannabinoid use expands, there grows a need to evaluate cannabidiol (CBD) and cannabinol (CBN) for toxicity potential. This study set out to meet this need using the Emulate human Quad-Culture Liver-Chip to evaluate these two compounds for hepatotoxicity potential.

Communications Medicine, part of Nature Portfolio (2022)

KEY Takeaways

• 870 Liver-Chips were used to evaluate the toxicity of 27 known hepatotoxic and non-toxic drugs.
• The Liver-Chip outperformed spheroids and animal models, with a sensitivity of 77% and specificity of 100% against the full set of 27 drugs tested on a single donor.
• Across a subset of 18 drugs tested on two donors, the Liver-Chip yielded a sensitivity of 87% and specificity of 100%.
• Economic analysis indicates routine use of the Liver-Chip could generate $3 billion per year to small-molecule drug development through an increase in R&D productivity​.

Abstract

Conventional preclinical models often miss drug toxicities, meaning the harm these drugs pose to humans is only realized in clinical trials or when they make it to market. This has caused the pharmaceutical industry to waste considerable time and resources developing drugs destined to fail. Organ-on-a-Chip technology has the potential improve success in drug development pipelines, as it can recapitulate organ-level pathophysiology and clinical responses; however, systematic and quantitative evaluations of Organ-Chips’ predictive value have not yet been reported. 

Webinar Abstract

In vivo gene therapy holds enormous promise to treat inherited and acquired genetic diseases affecting the liver, including lysosomal storage disorders, liver metabolic disorders, and hemophilia. Creating an effective transgene for these genetic disorders is only half the battle—it’s just as critical to create a safe and efficient vehicle to deliver the transgene to the target cells.  

The development of these vehicles is where one of the biggest challenges lies and where progress has been hampered due to limitations of conventional research models. Animal studies are time consuming, costly, and tightly regulated, while 2D cell culture models lack the complexity to deliver physiological relevance. These challenges have led to a slow pace of gene therapy development for safe and effective therapeutics to become available for patients in need. 

Instead of waiting months to get results from animal studies that may not translate to humans, it is now possible to get physiologically relevant data within weeks by using the Emulate Liver-Chip to assess gene therapy vectors. The adeno-associated virus (AAV) transduction application for the Emulate Liver-Chip enables gene therapy researchers to test the delivery efficiency and toxicity of AAV vectors in the most human-relevant in vitro model of the liver sinusoid, with proven validity in predicting drug toxicity. This AAV transduction application will allow rapid iteration on AAV design to optimize the delivery of gene therapies and accelerate therapeutic development.

In this webinar, you will learn how the Liver-Chip has been used to: 

• Assess time- and concentration-dependent AAV transduction efficiency 
• Discriminate between the transduction efficiency of different AAVs 
• Evaluate the toxicity of AAV-based gene therapy vectors 
• Study AAV transport from vasculature to target epithelial tissue in a proof-of-concept study 

Organ Model: Liver

Applications: Immunology & Inflammation

Highlights

Imaging endothelial cell behavior under physiological conditions, particularly those associated with chronic fibrotic pathologies, is an incredibly challenging endeavor. While short-term assessments (hours) can be achieved with techniques such as intravital microscopy, vascular changes often occur over days and weeks which is unfeasible with current imaging techniques. These challenges are exemplified within the liver where liver sinusoidal endothelial cells (LSECs) are known to undergo dramatic changes termed endothelial-to-mesenchymal transition (EndMT) during fibrotic liver disease. Despite the established presence of EndMT in liver disease, the inaccessibility of viable liver tissue, and simplicity of 2D culture techniques has meant, the role of EndMT during disease progression remains largely undetermined. This study describes the development of novel fluorescent EndMT reporters to identify, track, and characterize the migratory behaviour of EndMT cells. The authors show that liver-on-a-chip (LOAC) platforms provide a flexible, optically accessible, and physiologically relevant microenvironment to study the vascular dynamics of EndMT during liver disease.

Products Used In This Publication

Overview

In this Application Note for adeno-associated virus (AAV) transduction, learn how researchers can test the delivery efficiency and safety of AAV vectors using the Emulate Liver-Chip.

Key highlights:

• Assess time- and concentration-dependent AAV transduction efficiency  
• Discriminate between the transduction efficiencies of different AAVs  
• Evaluate the toxicities of AAV-based gene therapy vectors  
• Study AAV transport from vasculature to target epithelial tissue in a proof-of-concept study  

Organ Model: Liver

Application: Toxicology

Abstract: Drug-induced liver injury (DILI) is a significant public health issue, but standard animal tests and clinical trials sometimes fail to predict DILI due to species differences and the relatively low number of human subjects involved in preapproval studies of a new drug, respectively. In vitro models have long been used to aid DILI prediction, with primary human hepatocytes (PHHs) being generally considered the gold standard. However, despite many efforts and decades of work, traditional culture methods have been unsuccessful in either fully preserving essential liver functions after isolation of PHHs or in emulating interactions between PHHs and hepatic nonparenchymal cells (NPCs), both of which are essential for the development of DILI under in vivo conditions. Recently, various liver-on-a-chip (Liver-Chip) systems have been developed to co-culture hepatocytes and NPCs in a three-dimensional environment on microfluidic channels, enabling better maintenance of primary liver cells and thus improved DILI prediction. The Emulate Liver-Chip is a commercially available system that can recapitulate some in vivo DILI responses associated with certain compounds whose liver safety profile cannot be accurately evaluated using conventional approaches involving PHHs or animal models due to a lack of innate immune responses or species-dependent toxicity, respectively. Here, we describe detailed procedures for the use of Emulate Liver-Chips for co-culturing PHHs and NPCs for the purpose of DILI evaluation. First, we describe the procedures for preparing the Liver-Chip. We then outline the steps needed for sequential seeding of PHHs and NPCs in the prepared Liver-Chips. Lastly, we provide a protocol for utilizing cells maintained in perfusion culture in the Liver-Chips to evaluate DILI, using acetaminophen as an example. In all, use of this system and the procedures described here allow better preservation of the functions of human primary liver cells, resulting in an improved in vitro model for DILI assessment. © 2022 Wiley Periodicals LLC. This article has been contributed to by US Government employees and their work is in the public domain in the USA. Basic Protocol 1: Liver-Chip preparation Basic Protocol 2: Seeding primary human hepatocytes and nonparenchymal cells on Liver-Chips Basic Protocol 3: Perfusion culture for the study of acetaminophen-induced liver injury.

Webinar Abstract

The liver is responsible for the internalization and catabolic clearance of therapeutic antibodies as well as antibody-bound immune complexes. Liver sinusoidal endothelial cells (LSECs) are key actors in these processes as they express scavenging receptors that recognize, bind, and internalize an enormous diversity of extracellular ligands.

Investigating the pharmacological effects of antibody clearance via human liver has been challenging to do in vitro due to, among other issues, the lack of reliable long-term LSEC culture protocols. In response to these issues, the Emulate Liver-Chip was developed as an effective in vitro model capable of elongating LSEC reliability, allowing scientists to study antibody clearance like never before.

In this webinar, you will:  

• Learn how the Emulate Liver-Chip was used to recreate the liver microenvironment and extend the viability and function of LSECs, including CD32B expression levels, for a duration relevant for assessing the pharmacokinetics (PK) of therapeutic antibodies.
• Hear about results that show that the expression of CD32B can differ based on experimental variables such as the source of primary cells (donor), passage number or source of detection antibodies used to visualize CD32B, and shear stress. 
• Understand how the CD32B expression was maintained for 14 days on the Liver-Chip in a donor-dependent but passage number independent manner and
• See how the Scanning Electron Microscopy (SEM) imaging showed the presence of fenestrae structures—one of the hallmarks of LSEC function. 

Organ Model: Liver

Applications: Toxicology

Highlights

• Liver-Chip accurately predicted toxicity with most chemicals.
• Liver-Chip platform had high sensitivity and specificity, low variability, and high power.
• Liver-Chips and 24 well plates exhibited similar toxicity with many compounds.
• Flow of media or addition of LSECs may aid in slower toxicity seen in Liver-Chips.

Products Used In This Publication

Webinar Abstract

In this webinar, Dr. Jonathan Sexton presents an evaluation of iPSC-derived human liver organoids (HLOs) that spontaneously produce autologous hepatocytes, stellate, and Kupffer cells for drug-induced liver injury (DILI) risk prediction. The study leveraged Emulate Organ-Chips while using a multi-omics approach that integrated metabolomics, single-cell RNA sequencing, and high-content imaging to predict DILI risk with imputation of the mechanism of action. HLOs on the Liver-Chip were shown to dramatically increase albumin production and CYP450 expression while releasing ALT/AST when treated with drugs known to cause DILI at clinically relevant concentrations. Furthermore, HLO Liver-Chips were able to be used to evaluate inarigivir for hepatitis B by predicting the hepatotoxicity of the tenofovir-inarigivir combination that was responsible for unanticipated liver injury and death in a phase-III clinical study. This combination caused steatosis and mitochondrial perturbation in HLOs that recapitulate the clinical and histological presentation of the liver injury with a mechanism similar to fialuradine. 

The study “A Multi-Omics Human Liver Organoid Screening Platform for DILI Risk Prediction” is available to read on bioRxiv.