ORGANS-ON-CHIPS

organs on chip presentation

1. ORGANS-ON-CHIPS Eficacia y actividad in vivo: Ensayos de farmacocinética & bioanálisis SELECCIÓN DE DIANAS MOLECULARES Y VALIDACIÓN DE FÁRMACOS Jueid el Mahdi Carmen Luque Álvarez Eder Fernández Recio Máster de Biología Molecular Aplicada a Empresas Biotecnológicas ( BioEnterprise ) Universidad de Granada

2. INDEX INTRODUCTION ORGAN ON CHIP ADME PROCESS INTESTINAL-ON-CHIP KIDNEY-ON-CHIP LIVER-ON-CHIP LINKED ORGAN MPS FUTURE PROSPECTS CONCLUSIONS BIBLIOGRAPHY

3. Introduction Drug Testing in animal models is time consuming, costly and often does not predict the adverse effects in humans and also 60% of animal models are not able to predict the toxicity 3D Culture models have recently garnered great attention . They promote levels of cell differentiation not possible in conventional 2D Culture systems. 3D Culture systems is a large microfabrication technologies from the microchip industry and microfluidics. It approaches to create cell culture microenvironments This technology supports both tissue differentiation and recapitulate the tissue-tissue interfaces, spatiotemporal micro gradients and mechanical microenvironments of living organs. This organs on chips permit the study of human physiology in an organ specific context, enable novel in vitro disease models, and could potentially serve as replacements of animals used in drug development and toxin testing

4. This technology is being used to develop a whole pipeline of human organs on a chip such as lung, gut, liver, heart, skin, bone marrow, pancreas, kidney, eye and even a system that mimics the blood brain barrier. The idea is to recreate the smallest functional unit of any particular organ in a microenvironment that closely imitates the human body.

5. What is Organ On chip ? It is a multichannel 3-D micro fluidic cell culture chip which simulates the activities, mechanisms, physiological response of entire organs. These micro devices are translucent , they provide a window to watch inner workings of human organs

6. Steps of drug development using in vitro Microphysiological systems Estimation Of ADME parameters

7. Intestinal models on chip for absorption and first-pass metabolism. SYSTEMIC BIOAVAILABILITY

8. Intestinal on Chip Caco 2 cells form a polarized monolayer which are used to assess cellular permeability and active  vs.  passive uptake mechanisms. MDCK cells ,   is a more common cellular model since it does not require long culture times.  Neither model possesses representative  in vivo  CYP and transporter activity. Moreover, these are static 2D systems that do not possess many of the physiologic features seen  in vivo  such as mucus secretion, microbiome and peristalsis

9. Intestinal model development

10. What do we need from an intestinal-on-chip? Currently non-evaluable parameters First-pass metabolism on transit through tissue layer . Can be adapted to gut – liver system Desired functionality of intestine Chip An appropriate barrier function ≈ in vivo The ability to sample easily from both the apical and basolateral sides Cell viability and architecture Verified asymmetric distribution of key transporter expression similar to in vivo Functional expression of relevant transporters Reproduce inhibition and/or induction of metabolic enzymes and transporters. Dynamic nature of intestinal lumen

11. The future Add value adds value to essays: Key drug transport mechanisms are active First-pass gut metabolism is active and scalable to the  in vivo  setting. Adaptive responses of multiple cell types in the model due to drug treatment and disease may be assessed. Representation of the ileum would also allow for incorporation of enterohepatic recirculation of bile acids. In addition, the presence of colon-derived epithelial cells would not only allow for the study of colonic absorption, but would also allow for testing of colon-targeted prodrugs, which is another specific ADME application

12. Kidney-on-a-chip for drug metabolism and excretion The total excreted = glomerular filtration +tubular secretion + reabsorption Kidney model future uses: Kidney models can also be evaluated for their suitability to study tissue exposure/accumulation questions in relation to renal toxicity. Downstream nephron regions should consider how to verify retention of those specific metabolic functions in order to accurately recapitulate the physiology in those areas In addition, controlling fluidity rates in MPS models can allow a more accurate simulation of kinetic / physiological conditions in the body. Existing kidney systems for ADME studies: Kidney microsomes can be used as a benchmark for metabolic enzyme activities , including CYPs , flavin-containing … Primary human renal proximal tubular (HRPT) cells have also been occasionally utilized .

13. What do we need from an kidney chip? Currently non-evaluable parameters Apical– basolateral transport Kidney-relevant metabolism Multi-drug interaction on transport / metabolism Desired functionality of kidney Chip Representative barrier function between blood and compartments Appropriate reuptake transport from intraluminal space into the blood compartment (apical to basolateral) Kidney-relevant metabolism function pH settings múltiples tipos de células o regiones del riñón representadas

14. The future of kidney chip The development is focused on creating a model to evaluate renal toxicity. Demostration of expression and activity of key enzymes and transporters in this system would be the next most logical step . Kidney chip could offer a more stable enzymatic and transporter expression, and desired tissue architecture allowing greater opportunity for quantitative renal clearance or metabolism evaluations. In addition, control of fluidic rates in MPS models can allow for more accurate simulation of kinetic/physiologic conditions in the body.

15. Liver models Clearance , metabolite identification , induction and transport

16. Liver - on -a-chip Liver is the main organ of clearance for many drugs due to its high metabolic enzyme capacity , physiological positioning as a barrier to systemic exposure of orally administered drugs and biliary excretion functionality One aim of microphysiological liver systems is to capture more of the in vivo complexity by creating a more physiologically relevant environment Researchers have shown more ADME-relevant activities, for example demonstration of CYP activities and the inducibility of CYP enzymes The implementation of hepatocyte–Kupffer cell co-cultures is a likely next step in the development of long-term liver MPS systems

17. Liver - on -a-chip platforms Beckwitt et al , 2018 Du et al , 2017

18. Commercially Available Liver -Chip ( Emulate , Inc., 2019) in vivo -relevant responses to known and tool compounds at clinically relevant concentrations and through the appropriate mechanisms of action. General Toxicity · Mechanistic Toxicity · Nutrient Metabolism

19. Linked organ MPS CREATING A HUMAN INTEGRATIVE PHARMACOLOGY PLATFORM

20. “Human- on -a-chip” Although isolated MPS are valuable for modelling certain tissue-specific functions , there exists non-linear, dynamic organ crosstalk , especially in the context of disease or toxicity , that cannot be captured by studying these processes in isolation Lower-order (2–4 tissues ) inter-linked MPS are more tractable and can be used to examine the secondary effects of drugs on (un) intended target tissues A multi-compartment in vitro system amenable to interrogation and measurement can help uncover mechanisms underlying complex drug actions at the local and systemic level Most of the multi-MPS studies have been liver-centric , designed to interrogate the metabolic interplays between the liver and various tissue types ( gut , kidney , pancreas , heart , tumour ). These proof - of -concept applications include first-pass metabolism , metabolic activation or inactivation of parent compounds and their effects on downstream tissues and tissue-crosstalk in disease modelling One of the most powerful applications of a multi-MPS model is to capture not only the direct effect of the drug on the target but also the effects of metabolites and downstream pharmacodynamics of the drug as it undergoes sequential modifications throughout different parts of the body Fowler et al , 2020

21. Effect of hepatic metabolism on cardiotoxicity in a multi-organ human-on-a-chip system McAleer et al , 2019 Oleaga et al , 2018

22. Temporal PD/PK Relationship Derived McAleer et al , 2019

23. Application in drug discovery McAleer et al , 2019

24. FUTURE PROSPECTS Limitations The construction material of the device ( eg , Polydimethylsiloxane , PDMS) can adsorb non- specifically and also absorb hydrophobic compounds , limiting cellular exposure to the drug and generation of metabolites ; The use of low cell number and media volume in some microfluidic systems may pose a challenge for analytical detection ; The inability to access all tissue compartments for the organ-specific drug / metabolite profile over time; The lack of a mathematical methodology to extract the PK parameters necessary for IVIVE; The use of cell lines , which lack the enzymes and transporters relevant for PK studies . CHALLENGES Allow the ability to collect sufficient samples over time for temporary drug quantification In many proposed studies involving organ-on-a-chip, the usefulness of these devices was indicated by an evaluation of easily observed and measured functions A practical in vitro model should be a system that can observe and record a variety of physiological responses to specific biological stimuli Drugs action during pregnancy cannot be studied To incorporate lentiviral-induced fluorescent biosensors for an automated readout of various cellular functions

25. conclusionS One of the most significant targets for research tools development is the human-on-a-chip to replace animal models in research test and pharmaceutical industry The human-on-a-chip tool offers the opportunities for growth through the incorporation of more tissues with proper function and without external aid A completely independent system requires that all tissues can adequately provide its physiological function This new technology will act as a bridge between conventional cell cultures and new standardized clinical trial procedures without using animal-based models.

26. BibliogrAPHY Abaci HE, Shuler ML: Human-on-a-chip design strategies and principles for physiologically based pharmacokinetics/pharmacodynamics modeling . Integrative Biology 2015, 7 (4):383-391. McAleer CW, Pointon A, Long CJ, Brighton RL, Wilkin BD, Bridges LR, Narasimhan Sriram N, Fabre K, McDougall R, Muse VP et al : On the potential of in vitro organ-chip models to define temporal pharmacokinetic-pharmacodynamic relationships . Scientific Reports 2019, 9 (1):9619. Fowler S, Chen WLK, Duignan DB, Gupta A, Hariparsad N, Kenny JR, Lai WG, Liras J, Phillips JA, Gan J: Microphysiological systems for ADME-related applications: current status and recommendations for system development and characterization . Lab on a Chip 2020, 20 (3):446-467.

27. COMPLEMENTARy BIBLIOGRAPHY Beckwitt CH, Clark AM, Wheeler S, Taylor DL, Stolz DB, Griffith L, Wells A: Liver ‘organ on a chip’ . Experimental Cell Research 2018, 363 (1):15-25. Du Y, Li N, Yang H, Luo C, Gong Y, Tong C, Gao Y, Lü S, Long M: Mimicking liver sinusoidal structures and functions using a 3D-configured microfluidic chip . Lab on a Chip 2017, 17 (5):782-794. Oleaga C, Riu A, Rothemund S, Lavado A, McAleer CW, Long CJ, Persaud K, Narasimhan NS, Tran M, Roles J et al : Investigation of the effect of hepatic metabolism on off-target cardiotoxicity in a multi-organ human-on-a-chip system . Biomaterials 2018, 182 :176-190. " Biouptake And Bioabsorption Via Caco-2 Permeability Assay | Brunswick Labs". 2019. Brunswick Labs. https://brunswicklabs.com/capabilities/bioassays/caco-2-permeability-assay/. Emulate, Inc., 2019. [image] Available at: <https://www.emulatebio.com/intestine-chip> [Accessed 15 May 2020]. Emulate, Inc., 2019. [image] Available at: <https://www.emulatebio.com/liver-chip> [Accessed 15 May 2020].

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