In this webinar from Drug Discovery Day 2024, Carly Strelez, PhD, discusses her work using Organ-on-a-Chip technology to better understand colorectal cancer progression and transform approaches to personalized medicine.
Carly Strelez, PhD
Manager, Organ-on-a-Chip Team
Lawrence J. Ellison Institute for Transformative Medicine
Organ Model: Intestine (Caco2)
Application: ADME-Tox
Abstract: Microphysiological systems (MPSs) are promising in vitro technologies for physiologically relevant predictions of the human absorption, distribution, metabolism, and excretion (ADME) properties of drug candidates. However, polydimethylsiloxane (PDMS), a common material used in MPSs, can both adsorb and absorb small molecules, thereby compromising experimental results. This study aimed to evaluate the feasibility of using the PDMS-based Emulate gut-on-chip to determine the first-pass intestinal drug clearance. In cell-free PDMS organ-chips, we assessed the loss of 17 drugs, among which testosterone was selected as a model compound for further study based on its substantial ad- and absorptions to organ chips and its extensive first-pass intestinal metabolism with well-characterized metabolites. A gut-on-chip model consisting of epithelial Caco-2 cells and primary human umbilical vein endothelial cells (HUVECs) was established. The barrier integrity of the model was tested with reference compounds and inhibition of drug efflux. Concentration-time profiles of testosterone were measured in cell-free organ chips and in gut-on-chip models. A method to deduce the metabolic clearance was provided. Our results demonstrate that metabolic clearance can be determined with PDMS-based MPSs despite substantial compound loss to the chip. Overall, this study offers a practical protocol to experimentally assess ADME properties in PDMS-based MPSs.
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.
Organ Model: Intestine (Duodenum)
Application: Metabolic Disease
Abstract:
Sulfadoxine-pyrimethamine (SP) antimalarial therapy has been suggested to potentially increase the birth weight of infants in pregnant women in sub-Saharan Africa, independently of malarial infection. Here, we utilized female intestinal organoid-derived cells cultured within microfluidic Organ Chips to investigate whether SP could directly impact intestinal function and thereby improve the absorption of essential fats and nutrients crucial for fetal growth.
Using a human organ-on-a-chip model, we replicated the adult female intestine with patient organoid-derived duodenal epithelial cells interfaced with human intestinal endothelial cells. Nutrient-deficient (ND) medium was perfused to simulate malnutrition, resulting in the appearance of enteric dysfunction indicators such as villus blunting, reduced mucus production, impaired nutrient absorption, and increased inflammatory cytokine secretion. SP was administered to these chips in the presence or absence of human peripheral blood mononuclear cells (PBMCs).
Our findings revealed that SP treatment effectively reversed multiple intestinal absorptive abnormalities observed in malnourished female Intestine Chips, as validated by transcriptomic and proteomic analyses. SP also reduced the production of inflammatory cytokines and suppressed the recruitment of PBMCs in ND chips.
Our results indicate that SP could potentially increase birth weights by preventing enteric dysfunction and suppressing intestinal inflammation. This underscores the potential of SP as a targeted intervention to improve maternal absorption, subsequently contributing to healthier fetal growth. While SP treatment shows promise in addressing malabsorption issues that can influence infant birth weight, we did not model pregnancy in our chips, and thus its usefulness for treatment of malnourished pregnant women requires further investigation through clinical trials.
Organ Model: Intestine (Duodenum)
Application: ADME-Tox
Abstract: Microphysiological systems (MPS) incorporating human intestinal organoids have shown the potential to faithfully model intestinal biology with the promise to accelerate development of oral prodrugs. We hypothesized that an MPS model incorporating flow, shear stress, and vasculature could provide more reliable measures of prodrug bioconversion and permeability. Following construction of jejunal and duodenal organoid MPS derived from 3 donors, we determined the area under the concentration-time (AUC) curve for the active drug in the vascular channel and characterized the enzymology of prodrug bioconversion. Fosamprenavir underwent phosphatase mediated hydrolysis to amprenavir while dabigatran etexilate (DABE) exhibited proper CES2- and, as anticipated, not CES1-mediated de-esterification, followed by permeation of amprenavir to the vascular channel. When experiments were conducted in the presence of bio-converting enzyme inhibitors (orthovanadate for alkaline phosphatase; bis(p-nitrophenyl)phosphate for carboxylesterase), the AUC of the active drug decreased accordingly in the vascular channel. In addition to functional analysis, the MPS was characterized through imaging and proteomic analysis. Imaging revealed proper expression and localization of epithelial, endothelial, tight junction and catalytic enzyme markers. Global proteomic analysis was used to analyze the MPS model and 3 comparator sources: an organoid-based transwell model (which was also evaluated for function), Matrigel embedded organoids and finally jejunal and duodenal cadaver tissues collected from 3 donors. Hierarchical clustering analysis (HCA) and principal component analysis (PCA) of global proteomic data demonstrated that all organoid-based models exhibited strong similarity and were distinct from tissues. Intestinal organoids in the MPS model exhibited strong similarity to human tissue for key epithelial markers via HCA. Quantitative proteomic analysis showed higher expression of key prodrug converting and drug metabolizing enzymes in MPS-derived organoids compared to tissues, organoids in Matrigel, and organoids on transwells. When comparing organoids from MPS and transwells, expression of intestinal alkaline phosphatase (ALPI), carboxylesterase (CES)2, cytochrome P450 3A4 (CYP3A4) and sucrase isomaltase (SI) was 2.97-, 1.2-, 11.3-, and 27.7-fold higher for duodenum and 7.7-, 4.6-, 18.1-, and 112.2-fold higher for jejunum organoids in MPS, respectively. The MPS approach can provide a more physiological system than enzymes, organoids, and organoids on transwells for pharmacokinetic analysis of prodrugs that account for 10% of all commercial medicines.
Organ Model: Intestine (Caco2)
Application: Immune Cell Recruitment
Abstract: Over 2 million people in North America suffer from inflammatory bowel disease (IBD), a chronic and idiopathic inflammatory condition. While previous research has primarily focused on studying immune cells as a cause and therapeutic target for IBD, recent findings suggest that non-immune cells may also play a crucial role in mediating cytokine and chemokine signaling, and therefore IBD symptoms. In this study, we developed an organ-on-a-chip co-culture model of Caco2 epithelial and HUVEC endothelial cells and induced inflammation using pro-inflammatory cytokines TNF-α and IFN-γ. We tested different concentration ranges and delivery orientations (apical vs. basal) to develop a consistently inducible inflammatory response model. We then measured pro-inflammatory cytokines and chemokines IL-6, IL-8, and CXCL-10, as well as epithelial barrier integrity. Our results indicate that this model 1. induces IBD-like cytokine secretion in non-immune cells and 2. decreases barrier integrity, making it a feasible and reliable model to test the direct actions of potential anti-inflammatory therapeutics on epithelial and endothelial cells.
Featured session at Netherlands MPS Day, which took place on November 15, 2023.
Joram Mooiweer from the University Medical Center Groningen presented his team’s work on modeling celiac disease using advanced human-relevant systems, including intestinal organoids and Organ-Chips. Celiac disease involves a complex interplay of genetic predisposition, immune responses (especially from tissue-resident T cells), and interactions with the intestinal epithelium and microbiota. By taking advantage of patient-derived tissue and induced pluripotent stem cells (iPSCs), the group aims to reconstruct these intricate processes in vitro.
Mooiweer highlights two approaches to modeling celiac disease. First, using patient biopsies, they isolate both intraepithelial lymphocytes (IELs) and intestinal stem cells. Culturing organoids derived from these stem cells together with IELs in a carefully optimized co-culture medium, they can recreate aspects of the cytotoxic activity associated with celiac disease—something that can be further manipulated using bispecific antibodies. These co-cultures maintain viability of both the epithelial and immune cells, enabling exploration of celiac-related immune-mediated epithelial damage.
Second, the group also uses iPSCs from patients and controls to generate small intestine-like epithelial tissues that self-organize into complex, 3D folded architectures when seeded on chip. With the help of growth factor gradients, they achieve a cell-type composition and maturation state that closely mirrors the human small intestine, including various specialized epithelial cells and mesenchymal cell types. This system is genetically tractable and can be further refined to incorporate inflammatory cues, immune cells, and eventually patient-specific microbiota under physiologically relevant conditions.
Ultimately, these multifaceted in vitro platforms will enable mechanistic insights into how genetic variants influence celiac disease onset and progression. They also provide a foundation for modeling interactions between epithelial cells, immune cells, and environmental triggers, paving the way for more predictive and personalized therapeutic strategies.
Key learnings from this presentation include:
Organ Model: Intestine (Caco2)
Application: Model Development
Abstract: Although mutations in the SCRIB gene lead to multiple morphological organ defects in vertebrates, the molecular pathway linking SCRIB to organ shape anomalies remains elusive. Here, we study the impact of SCRIB-targeted gene mutations during the formation of the gut epithelium in an organ-on-chip model. We show that SCRIB KO gut-like epithelia are flatter with reduced exposed surface area. Cell differentiation on filters further shows that SCRIB plays a critical role in the control of apical cell shape, as well as in the basoapical polarization of myosin light chain localization and activity. Finally, we show that SCRIB serves as a molecular scaffold for SHROOM2/4 and ROCK1 and identify an evolutionary conserved SHROOM binding site in the SCRIB carboxy-terminal that is required for SCRIB function in the control of apical cell shape. Our results demonstrate that SCRIB plays a key role in epithelial morphogenesis by controlling the epithelial apical contractility during cell differentiation.
Oncology drug candidates are currently the least likely type of therapeutic to succeed in clinical trials, with only 5.1% of Phase I candidates going on to receive FDA approval1. Understanding a tumor’s microenvironment is key to regulating cancer progression and developing more effective therapies—and Chip-A1 will give researchers this capability.
In this August 15, 2023 webinar, Luke Dimasi, Senior Director of Product Management at Emulate, provided an overview of the Chip-A1 Accessible Chip, a new Organ-Chip consumable that expands the applications of Organ-on-a-Chip technology by allowing users to create thicker, multilayered tissues within the epithelial culture chamber and directly treat the tissue with topical or aerosolized drugs. Following this introduction, Elee Shimshoni, PhD, postdoctoral researcher at MIT, discussed how she and her former team from the Wyss Institute at Harvard used a prototype of Chip-A1 and found that it offered a new approach for studying epithelial-stromal interactions in Barrett’s Esophagus as well as the broader underlying mechanisms associated with esophageal cancer progression.
During this webinar, the speakers discussed:
About Organ-on-a-Chip Technology
Organ-on-a-Chip technology is poised to deliver a paradigm shift in drug discovery. By emulating human physiology, Organ-Chips have the potential to increase the predictive power of preclinical modeling and advance more drugs to the clinic. Learn more about Organ-on-a-Chip technology by downloading our free eBook.
With the potential to bring cures to some of the world’s most devastating diseases, including cancer, muscular atrophy, and blindness, gene therapy is a cutting-edge area of research that is generating a tremendous amount of excitement. Download this free eBook to learn what gene therapy is, how it works, and why Organ-on-a-Chip technology could help gene therapy reach widespread use.
