Transcript PLASMA POSTER COMPETITION

PLASMA POSTER
COMPETITION
All Staff and Students are invited to
use the ballot papers below to vote for
your favourite plasma poster


Jennifer Caffarel
1/3 of men
will have
bothersome urinary symptoms
at some point in their life
… then you might have
“Bladder Outlet
Obstruction”
The prostate gland
surrounds the
urethra
With increasing age,
it increases in size
and can obstruct the
urethra…
… and can cause a
slow or “weak”
urine flow.
Urine flow can be quantified
by a technique called
“uroflowmetry”.
Uroflowmetry is performed in
clinics, using expensive
electronic flowmeters
£4
0
0
0
Attending a flow clinic can be
a long and tedious
experience…
… and having to fill and empty
your bladder several times in a
short period of time is rather
unnatural!
The data obtained from flow clinics is
variable, due to a number of factors:
• Time of day
• Volume of urine in the bladder
• Emotional state of patient
The aim is to improve the diagnosis
made using uroflowmetry.
Electronic
Flowmeter
0
£
0
40
vs.
£5
Basic
Flowmeter
For example, we tested a very basic
flowmeter, to obtain more
representative results.
This is how the basic flowmeter is
used:
• The patient takes
the device home
• Makes multiple
measurements over
several days, in his
own time
• This home flowmeter provided
more representative results than inclinic flows.
• And 2 patients who could not
urinate in the clinic were able to
use this device.
PLASMA POSTER
COMPETITION
All Staff and Students are invited to
use the ballot papers below to vote for
your favourite plasma poster
PLASMA POSTER
COMPETITION
All Staff and Students are invited to
use the ballot papers below to vote for
your favourite plasma poster
?
?
?
?
?
?
?
?
Questions (and some answers) on
? research into stomach cancer…
?
?
?
Claire Worrall
?
?
?
?
?
?
Who
gets stomach cancer?
It is difficult to predict.
It depends on:
 If you have H. pylori infection
 What you eat
 If you have stomach ulcers
 How much alcohol you drink
 If you smoke
 Your Body Mass Index
 What country you live in
 Your genetic background
 Your gender
So biological markers of early disease would be very useful.
What
changes occur in stomach cancer?
100 %
100.00%
Expression levels of many
proteins are altered…
90.00%
80.00%
% of cases
expressing
TFIZ1
70.00%
60.00%
20.00%
10.00%
0%
0.00%
Normal
stomach
Stomach
cancer
0.005 %
MKN74
KATO III
SNU1
SNU5
SNU 16
NUGC3
SNU 5
AGS
SNU 1
0%
KATO III
% of normal
TFIZ1
expression
MKN74
It is virtually undetectable by
RT-PCR in our panel of 7
gastric cancer cell lines.
30.00%
NUGC3
TFIZ1 is expressed in mucinsecreting cells and secreted
into the mucous layer in the
stomach.
40.00%
AGS
We are studying one protein
whose expression is lost in
the majority of stomach
cancers:
50.00%
SNU16
Where
can we study stomach cancer?
There is limited supply of patient
samples.
However, we can grow stomach cancer
cells in the lab to perform tests on.
These cells grow and divide, allowing us
to measure the effects of different
conditions on:
How fast they grow and divide
How easily they die
How quickly they move
These behaviours in patient cancer cells help determine
tumour growth and the ability to metastasize.
When
do we study a protein in stomach cancer?
When we have evidence linking it to stomach cancer…
TFIZ1 is bound in the stomach to TFF1, which has
many links to cancer:
High expression
in oestrogen responsive
breast cancer
TFF1
TFIZ1
Involved in
ulcer repair
30% null mice get
stomach cancer
Binds to H. pylori
TFF1
TFF1
How
are we studying this protein?
By introducing controllable TFIZ1 expression into
stomach cancer cells:
1. Transfect in
regulator plasmid
neoR
tTA
TFIZ1
Off
TFIZ1
On
pTet-Off
The cells now
express TFIZ1
TRE
2. Transfect in
TFIZ1 plasmid
TFIZ1
pTRE-Tight TFIZ1
neoR tTA
TRE TFIZ1
Stomach cancer
Cell line
Adding
doxycycline
turns off TFIZ1
expression
Why
do we study a protein in gastric cancer?
If we know
WHY the expression changes
WHAT the protein does
In the normal stomach
In stomach cancer
WHEN the expression changes
It can
Help us decide whether it might be a good early indicator of cancer.
Help us decide whether it might be a good therapeutic target.
Acknowledgements:
 Felicity May
 Herbie Newell
 Northern Institute for Cancer Research
Claire Worrall
[email protected]
PLASMA POSTER
COMPETITION
All Staff and Students are invited to
use the ballot papers below to vote for
your favourite plasma poster
INTRODUCTION:
[email protected]
‘MRI scan of the breast reveals two
discrete areas of abnormality, which
proved to be cancer.’
Chemotherapy and
radiotherapyinduced
second cancers are the leading cause of death in
patients cured of Hodgkin Lymphoma (cancer of
the lymphatic system). Young women treated for
Hodgkin Lymphoma with radiotherapy at the
chest are at a high risk of developing breast
cancer
AIM:
To develop an in vitro model system of
radiogenic breat cancer, so that the molecular
genetic events associated with transformation
and the development of a radio-resistant
phenotype can be investigated.
PROCEDURE:
1.
This project will involve exposing
transformed (MCF-7) and non-transformed
(MCF-10) breast cell lines to increasing
doses of ionising radiation.
2.
Post treated cell lines will be compared to
untreated parental cells across the whole
genome to identify regions characterised
by loss of heterozygosity, copy number
alterations and uniparental disomy.
3.
Candidate locations of interest will be
further investigated in treated cell lines
and in breast tissue from patients with
breast cancer following radiotherapy for
Hodgkin Lymphoma.
http://www.breastcancer.org/pictures/diagnosi
s/mammogram/mri_2.jsp
FUNDED BY: Northern Institute for
Cancer Research
PLASMA POSTER
COMPETITION
All Staff and Students are invited to
use the ballot papers below to vote for
your favourite plasma poster
Forkhead Proteins & Cancer
Frances Purtill
iCAMB
Forkhead Proteins & Cancer
Unregulated cell proliferation is a
characteristic of all cancers
Understanding the mechanisms
behind cell proliferation will help in
the treatment and prevention of
cancer
Forkhead Proteins & Cancer
Cell proliferation is controlled by a series of events
known as the Cell Cycle
The cell cycle consists of 4 phases:
3&4
G1 and G2 are
‘gap’ phases
which prepare
the cell for its
next step
2
M Phase (Mitosis)
Replicated DNA is
equally distributed
into 2 cells
1
S Phase
DNA is replicated
M
G2
G1
S
Genetic material is copied and 2 identical cells
are formed
Forkhead Proteins & Cancer
Forkhead Transcription Factors (FKH-TF) are critical
for the regulation of the cell cycle
Forkhead
proteins
They
bind upstream
of genes have
involvedbeen
in
mitosis, and activate their expression
associated with various human
FKH-TF
cancers
GENE SWITCHED ON
TARGET GENE
TARGET PROTEIN MADE
Forkhead Proteins & Cancer
Forkhead
proteins
for mitosis
in all
Therefore
it isarea vital
valuable
model
eukaryotes, from yeast to man
to study forkhead proteins, and
Yeast
is easy toour
manipulate,
genetically
and
improve
knowledge
of the
biochemically
cell cycle in humans
In the future, this knowledge
could potentially be used to
improve existing cancer therapies
PLASMA POSTER
COMPETITION
All Staff and Students are invited to
use the ballot papers below to vote for
your favourite plasma poster
The Development of Small Molecule
Inhibitors of the MDM2-p53 Interaction
Junfeng Liu
p53 is a Tumour Suppressor
% mice with tumour
p53-/-
74% at 6 months
p53+/-
2% at 9 months
p53+/+
1% at 18 months
Donehower et al. Nature, 1992
Also suggest to read Donehower et al. Nature genetics, 1993
MDM2 controls p53 through an autoregulatory negative-feedback loop
p53
p53
MDM2
p53’s proteasomal
degradation
MDM2
mdm2 mRNA
mRNA of other targets
Three mechanism to repress p53 by activation of MDM2-expression:
1.
MDM2 binds p53 at transactivation domain and blocks its ability to activate transcription
2.
MDM2 acts as an E3 ubiquitin ligase that promotes p53’s proteasomal degradation
3.
MDM2 is involved in the nuclear export of p53
Breaking the negative-feedback loop
with antagonists
p53
p53
MDM2
p53’s proteasomal
degradation
MDM2
mdm2 mRNA
mRNA of other targets
 Therapeutic potential
Rescue of p53 Function by Disrupting the p53–MDM2 Interaction.

Blocking MDM2 Expression

Inhibiting MDM2 Ubiquitin Ligase Activity

Disruption of the p53–MDM2 binding
Possible Methods
 Disruption of the p53–MDM2 Binding
Interaction
 Antibody microinjection
 Peptide analogues corresponding to the MDM2
binding domain of p53
 Small molecular weight compounds
The p53-MDM2 interaction:
Unique binding site
Unique
& Good for Small
Molecule Drug Design
and Development
Structure based drug design
MDM2
Binding
Domain
(Binding area
high lighted)
PyMOL Software were applied for the followed cartoons.
MDM2 protein and p53 peptide structures come from Protein Data Bank.
Structure based drug design
8mer p53
peptide
Binding
with MDM2
Structure based drug design
Three key
residues of
p53 in the
hydrophobic
pocket of
MDM2
Structure based drug design
Three key
residues of
p53 were
picked out for
drug design
Structure based drug design
Drug design
based on the
three key
residues of
p53
Structure based drug design
Designed small
molecule scaffold
Structure based drug design
Manual
docking of the
compound with
MDM2 binding
domain
Inhibitor Screen Using ELISA
Biotinylated
IP3 Peptide
M
M
M
Addition of:
Y Y Y
M
M
M
M
Incubation of MDM2
with antagonist
Incubation with
MDM2
Y
M
M
MDM2 primary antibody,
HRP conjugated
secondary antibody
M
ECL detection
ECL detection
LIGHT
M
YY
M
M
M
Streptavidin
DECREASED
LIGHT
Cellular activity Confirmation via
Western Blotting Assay
Effect of MDM2 inhibitors on the cellular levels of p53, MDM2 and p21
Nutlin-3
DMSO 0.5
1
10
NU8XXX
1
10
20
mM
MDM2
p53
p21
Actin
Thank you
for
your attention!
PLASMA POSTER
COMPETITION
All Staff and Students are invited to
use the ballot papers below to vote for
your favourite plasma poster
…and tools to diagnose Lower
Urinary Tract Dysfunction
1 in 3 men will suffer urinary
symptoms in their life:
struggling to begin pain while peeing
peeing
inability
to
pass
any
having to rush to
water
the toilet
Incontinence
having to go often
Poor stream
having to go in the ‘Terminal dribble’
middle of the night
Incomplete emptying
To diagnose the cause we
need to measure:
•Bladder pressure
•Urine flow rate
•Volume of urine voided
… among other tests
Well, how do you take a blood
pressure measurement?
How do you take a bladder
pressure measurement?
Use the same principle…
Place Pressure
a small,isinflatable
measuredcuff
around the penis…
Pressure
Flow rate
Volume
…and measure
the
pressure
to
Flow is interrupted
stop urine flow!
Penile Cuff Machine
Conventionally,
pressureWe can
can also
now use
use
We
itthis
to
flow
measurements
non-invasive
machine
measure
the bladder
would
require
urethral
clinically
to diagnose
characteristics
in
catheterisation,
an
Bladder people
Outlet to
healthy
uncomfortable
Obstruction
inprocedure
patients.
further
understand
the
with
risk of infection.
mechanics
of the lower
urinary tract.
With this tool we hope to:
• Verify theoretical bladder
models
• Understand better how the
bladder copes with obstruction
• Apply this to diagnosis and
treatment of patients
PLASMA POSTER
COMPETITION
All Staff and Students are invited to
use the ballot papers below to vote for
your favourite plasma poster
Heart Physiology: “The role of
p50”
Silvia Gaspar Pereira
PhD student with the Cell Signaling Group
Institute of Cellular Medicine
Light from the Heart Nebula - apod.nasa.gov
Heart Physiology: “The role of
p50”
What does p50 do?
When the cell receives a
signal, the protein p50 is
released from the cytoplasm
into the nucleus.
p5
0
p5
0
In the nucleus, p50 binds
DNA and thereby induces a
cellular response to the
signal.
In the absence of p50, this
signaling pathway is
impaired.
Aim: Is p50-dependent
signaling important in
heart muscle cells?
Heart Physiology: “The role of
p50”
To answer this question, we use mice lacking p50:
In mice without p50, hearts are
significantly bigger than in
normal mice.
p5
0
p5
0
The p50-deficient hearts are
more sensitive to certain
injuries, e.g. lack of
oxygenation in the tissue.
Future Work:
Does p50 help to prevent injury in the heart tissue?
Is p50 necessary during heart formation pre- or
postnatally?
If p50 is directly involved in these events, it could become
a potential target for heart disease treatments!
PLASMA POSTER
COMPETITION
All Staff and Students are invited to
use the ballot papers below to vote for
your favourite plasma poster
Regulation of the oxidative stress
response in the pathogenic fungi
Candida albicans
Miranda Patterson
Supervisor – Dr Janet Quinn
http://www.chuv.ch/imul/euresfun
Candida albicans
But, in immunocompromised people, such as
Candida
albicans
is the
major
systemic
people with
HIV/AIDS,
some
cancer
patients
pathogenic
fungi
of
humans
or
transplant
patients,
C.albicans
can cause
In immunocompetent people C.albicans
is the
life threatening
infections,
a
cause
of common systemic
superficial
infectionswith
such≈ as
rate
oral38%
andmortality
vaginal thrush
http://pathmicro.med.sc.edu/mycology/mycology-3.htm
http://pathmicro.med.sc.edu/mycology/mycology-3.htm
C.albicans invasion of blood vessels C.albicans invasion of a kidney from
in the oesophagus
an immunocompromised rabbit
Why are we interested in C.albicans
oxidative stress responses?
Neutrophils
But C.albicans
are important
can kill macrophages!
in killing C.albicans
Macrophages
andbyneutrophils
of the
Neutrophils kill
producing are
toxiccells
reactive
Macrophage
innate
immune
system
responsible
for
killing
oxygen
species
in
a
respiratory
burst
Full understanding of mechanisms used by
micro-organisms
C.albicans to overcome oxidative stresses
C.albicans C.albicans lacking key proteins involved in
C.albicans
Macrophage
may
reveal
novel
therapeutic
targets!
hyphae
oxidative stress responses are more sensitive to
www.biology.uark.edu/dmcnabb/dmcnabb.html
www.biology.uark.edu/dmcnabb/dmcnabb.html
killing by neutrophils and have reduced
C.albicans cells are virulenceAfter ingestion C.albicans
ingested by the
macrophage
starts growing hyphae and
escape the macrophage
What are we doing?
Oxidative
Stress
-ive
Cap1
Nucleus
Anti-oxidant
genes
Previous research shows
that following oxidative
Investigating the major
stress, Cap1 translocates to
oxidative
stress
pathway
in
We wantnucleus
to know whatand
is
the
C.albicans
– the
Cap1
regulating
Cap1?
upregulates transcription
pathway
of anti – oxidant genes so
that cells can survive
oxidative stress.
How are we doing this?
H2O2 concentration (mM)
Control
Wild type strain
Deleting
potential key regulators of Cap1
Cap1 delete
strain
Ybp1 delete
strainseeing how cells survive an oxidative
and
stress challenge
Wild type cells grow until high concentrations of H2O2
Have
weC.albicans
found
regulator
Cap1
atnothigh
Cells
lacking
Cap1
areasensitive
to H2of
02 and
grow
We
grow
in the presence
ofcan
hydrogen
even
concentrations
peroxide
(HHOat
tooxidative
induce
oxidative
stress
levels of
0) low
challenge?
2 22 2
Continuing
will hopefully
answer
Cells
lacking the research
potential regulator
of Cap1,
Ybp1, are
question!
also sensitive to H2this
02 but
only at high concentrations