Stem Cells in Drug Discovery

Meghma Mitra (RA1711014010082)

Vedant Rajesh Buwa (RA1711014010086)
Sabari Krishnan B. B. (RA1711014010088)
 Lesson Plan

Introduction to Stem Cells: Types and Sources

Applications of SCs in Drug Discovery

 What are Stem Cells?

Stem Cells (SCs) are special cells that have the ability to differentiate into many different cell types.

The 3 main types of SCs used in Drug Discovery:

❖ Pluripotent SCs (or embryonic SCs): SCs which can give rise to all cells except that of the
extraembryonic membranes. These cells differentiate into the ectoderm, mesoderm, and endoderm cell
lineages.
❖ Multipotent SCs (or adult SCs): SCs which can give rise to a few cell types. Eg: generally, hematopoietic
SCs give rise to RBCs, WBCs, and platelets, only.
❖ Induced Pluripotent SCs (iPSCs): terminally differentiated cells which are artificially engineered to
activate and express the genes which are responsible for pluripotency.
http://csmres.co.uk/cs.public.upd/article-downloads/Hook_2012_Drug-Discovery-Today.pdf
http://csmres.co.uk/cs.public.upd/article-downloads/Hook_2012_Drug-Discovery-Today.pdf
Sources of Stem Cells

❖ Embryonic SCs: from the inner cell mass of blastocyst (5-7 days after fertilization); collected
from IVF Clinic with ethical consent

❖ Adult SCs: from specific regions of adult organs; mainly the mesenchymal SCs. Eg: intestinal
stem cells are located in the crypts of Lieberkühn.
https://www.ebi.ac.uk/sites/ebi.ac.uk/files/content.ebi.ac.uk/materials/2014/140306_SME/humanising_drug_discovery_-_alex_gutteridge.pdf
Applications of SCs in Drug
Discovery
1. Target Identification
2. High-throughput Screening
3. Disease Modelling
4. Regenerative Drugs
5. Toxicology
6. Robotic High-throughput Platform
7. Microarray/Microfluidics Systems
8. Combinatorial Cell Culture

1. Target
Identification
Studies gene expression
patterns and dissect the
molecular mechanisms of
lineage commitment, thereby
identifying possible candidate
proteins for therapeutic
intervention.
● It involves the complete understanding of the molecular
mechanisms behind a biological process.
● The disease-causing protein(s) are targeted with therapeutic
small molecules, nucleic acids, or peptides to regulate its
function.
● Helpful in finding therapeutic strategies to diseases of in vivo
and even genetic origins.
● Zinc finger nucleases (ZNFs), transcription activator-like
effector nucleases (TALENs), Cre-LoXP system, and CRISPR-
tracrRNA system are some of the main gene editing systems
used to generate knockdown models to study disease targets.
 2. High-throughput
Screening
(HTS)
A powerful technique for
selecting the lineage of choice
from a heterogeneous mix of
differentiated cells. By this, the
effect of the drug used is also
established.
Overcomes the disadvantage of
using primary and recombinant
cell lines for target identification
● The disadvantage of using primary cell lines for HTS and
target identification is its relatively small number of
“satisfying” cells and donor-dependent variability, respectively.
● For HTS using SCs, sophisticated and sensitive instruments
and set-ups are required, which is a disadvantage.
● Drug discovery is performed similar to Target Identification,
but the only difference is instead of completely dealing with
the test genome, we analyze the effects of our drug of interest
in a model using highly sensitive screening techniques.
● Detecting techniques like multi-well plate ELISA and
microarray, imaging techniques like fluorometric plate readers
and photoproteins, sensors, and biochips are some of the
components using for HTS.
https://sci-hub.se/10.1089/adt.2010.0305
https://sci-hub.se/10.1016/j.copbio.2017.03.005

3. Disease
Modelling
An unavoidable part of any in
vivo study. Using various gene
editing and molecular
techniques, organismic models
are generated to study drug-
target interaction and thus drug
discovery.
● SCs can be used to generate cell line models which form the
basis of drug effectiveness testing.
● Usually done by molecular tools discussed in Target
Identification; either a gene is dysregulated or knocked-out.
● Cell line models are used for wide ranges of diseases, but
cannot explain the in vivo effect of drugs. This calls the need
for animal models, especially for diseases like PD, AD, ALS,
SMA, SCID and juvenile diabetes.
● SCs when administered as carriers of therapeutic gene to such
patients help in bringing a long-term cure.
https://sci-hub.se/10.1089/adt.2010.0305
 ● This is the most potent application of SCs, turning

4. Regenerative regenerative medicine a synonym for stem cell therapy.

Drugs ● Especially useful in disorders of degeneration, such as AD and

MDs, skin burn, ischemia and strokes.
Drugs which help in repopulating
diseased or lost cells. ● This is a readily available class of drugs with several
examples. Even drug repurposing show effectiveness.

● Eg.: Stem cells, especially mesenchymal SCs, were used for

screening 1000s of compounds for their ability to promote
osteogenesis.

● SCs are being used to screen small molecules that has

detrimental effects on the terminal differentiation of
progenitors into their respective cell types, which can be a
cause of many microenvironment-induced degenerative
disorders.
Stem Cells

5. Toxicology
The study of adverse effects of
chemical substances on living
organisms, and the practice of
diagnosing and treating
exposure to toxins and toxicants.
● Approximately 30% of drugs fail in early stage clinical trials
because of toxicity issues, primarily hepatic and cardiac
toxicity, causing loss of billions of dollars.
● Primary hepatocytes and cardiomyocytes are currently utilized
for toxicity studies, but when compared to SCs, they are very
difficult and expensive to manage.
● The use of iPSCs in toxicity studies revolutionized
personalized medicine; converting the required terminally-
differentiated cells of the donor into pluripotent stem cells,
help study the toxic effect of the drug individually, to some
extend… Toxicity of a drug, after all has a strong relation with
the genetic makeup of the individual!
 6. Robotic High- ● The platform integrates proprietary bioinformatics tools and

throughput automation to enable rapid rational screening of large

numbers of combinations of biological factors.
Platforms
● The culture medium, media supplements, cell adhesion
It is the approach of using
surface and culture vessel can all be optimized.
automated cell culture systems
to screen multiple differentiation
● Optimizing SC conditions can even mimic a 3D environment
conditions in multi-well format.
just like in our body, possibly overcoming the barrier between
in vitro and in vivo studies. We can save lots of lives, actually
of those poor animals!!!
https://www.institut-vision.org/en/facilities/8-platforms-institute/25-high-throughput-screening.html
7. Microarray or
Microfluidics
Systems
This approach has been used to
study the interplay between stem
cell fate and the
microenvironment, and propose
possible drugs which play a role
in it.
Microfluidics technology can
also be used to elucidate time
dependent processes enabling
rapid medium exchange and
culture condition switching at
desired time points.
● Different ECM and cell adhesion factors can be robotically
spotted onto microarrays in various combinations, enabling
screens of tens to hundreds of putative microenvironments.
● Microfluidics technology is an advanced HTS method which is
both automatic and time saving. We can run many
experiments at the same time using a small culture volume.
● An example of its intricacy: A multi-well microarray platform
enables 1200 simultaneous experiments on 240 unique
signaling environments. Each well contains 100 (20 mixtures
in five replicates) ECM spots in the context of a different
media composition. A reporter ES cell line (with a reporting
fluorophore) was used to monitor cell differentiation using a
confocal microarray scanner.
https://sci-hub.se/10.1039/C5LC01249J
 ● Combines miniaturisation of cell culture on microcarriers, a
8. Combinatorial pooling/splitting protocol and a unique tagging system to
Cell Culture enable multiplexing of experiments.

● Stem cells grown on microcarrier beads are shuffled randomly,

A technology to screen tens of
thousands of protocols in one
experiment.
stepwise through multiple differentiation media using a split-
pool method. This is done iteratively to increase the number of
experiments.

● Since large numbers of conditions can be tested in each

screen, it is possible to efficiently discover optimised
protocols that have advantages over more traditional cell
culture methods like serum-free, which use only small
molecules or exclude other variable and expensive products.
http://csmres.co.uk/cs.public.upd/article-downloads/Hook_2012_Drug-Discovery-Today.pdf
There’s a lot more
to be done...
If not us, then who will???
Thank you!
Any Questions?