organs on chip presentation

1. ORGANS-ON-CHIPS
EFICACIA Y ACTIVIDAD IN VIVO: ENSAYOS DE FARMACOCINÉTICA &
BIOANÁLISISSELECCIÓN 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
1. INTRODUCTION
2. ORGAN ON CHIP
3. ADME PROCESS
4. INTESTINAL-ON-CHIP
5. KIDNEY-ON-CHIP
6. LIVER-ON-CHIP
7. LINKED ORGAN MPS
8. FUTURE PROSPECTS
9. CONCLUSIONS
10. BIBLIOGRAPHY

3. INTRODUCTI
ON
• 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

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
ADME
DISTRIBUTION
METABOLISM
EXCRETION
ABSORPTION

7. •Can Occur Through :
•Gastrointestinal-Tract
•Lungs
•Skin
ABSORPTION
•Following Absorption the Drug is distributed to each organ.
• Parameters affecting distribution :
•Unbound Fraction
•The Drug Partitionning
•Blood to plasma Ratio
DISTRIBUTION
•Liver is the main organ
METABOLISM
EXCRECION

8. INTESTINAL MODELS ON CHIP FOR
ABSORPTION AND FIRST-PASS
METABOLISM.
Fraction
absorbed
(Fa)
•Physiochemical
properties
•Solubility
•Membrane
permeability
First-
pass
metabolis
m
•Metabolic reactions
•CYP
•Enzymas Phase 1 y 2
Hepatic
availability
(Fh)
SYSTEMIC
BIOAVAILABILITY

9. 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

10. INTESTINAL MODEL DEVELOPMENT
Epithelial cells from intestinal organoids derived
from pluripotent stem cells.
Use primary human intestinal cells or organoids,
instead of Caco-2 cells
Model 3D:
Cell monolayers to form villi-like structure
Co-culture Caco-2 cells with anaerobic bacteria
Culture multiple cells types along with mimicking
peristalsis
Mucus production
Model 2D:
Co-culture Caco-2 cells with the goblet-like cells
HT29-MTX
Development of a pumpless microfluidics system

11. 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

12. 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

13. 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.

14. 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

15. 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.

16. LIVER MODELS
CLEARANCE, METABOLITE IDENTIFICATION, INDUCTION AND TRANSPORT

17. 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

18. LIVER-ON-A-CHIP PLATFORMS
Beckwitt et al,
2018
Du et al,
2017

19. 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

20. LINKED ORGAN MPS
CREATING A HUMAN INTEGRATIVE PHARMACOLOGY PLATFORM

21. “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
Fowler et al,
2020

22. EFFECT OF HEPATIC METABOLISM ON
CARDIOTOXICITY IN A MULTI-ORGAN HUMAN-
ON-A-CHIP SYSTEM
McAleer et al,
2019
Oleaga et al,
2018

23. TEMPORAL PD/PK RELATIONSHIP DERIVED
McAleer et al,
2019

24. APPLICATION IN DRUG DISCOVERY
McAleer et al,
2019

25. 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;
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

26. 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.

27. BIBLIOGRAPHY
1. 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.
2. 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.
3. 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.

28. COMPLEMENTARY BIBLIOGRAPHY
1. 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.
2. 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.
3. 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.
4. "Biouptake And Bioabsorption Via Caco-2 Permeability Assay | Brunswick Labs". 2019.
Brunswick Labs. https://brunswicklabs.com/capabilities/bioassays/caco-2-
permeability-assay/.
5. Emulate, Inc., 2019. [image] Available at: <https://www.emulatebio.com/intestine-
chip> [Accessed 15 May 2020].
6. Emulate, Inc., 2019. [image] Available at: <https://www.emulatebio.com/liver-chip>
[Accessed 15 May 2020].

29. GRACIAS POR VUESTRA ATENCIÓN
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