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 menwill 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…

Urine flow can be quantified by a technique called “uroflowmetry”.

… and can cause a slow or “weak” urine flow.

Uroflowmetry is performed in clinics, using expensive electronic flowmeters

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.

For example, we tested a very basic flowmeter, to obtain more

representative results.

vs.Basic Flowmeter

Electronic Flowmeter

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 in-clinic flows.

• And 2 patients who could not urinate in the clinic were able to use this device.







Questions (and some answers) on

research into stomach cancer…

Claire Worrall

??

? ??

?

?

???

?

? ?

?

???

?

Who

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

gets stomach cancer?

So biological markers of early disease would be very useful.

Whatchanges occur in stomach cancer?

Expression levels of many proteins are altered…

We are studying one protein whose expression is lost in the majority of stomach cancers:

TFIZ1 is expressed in mucin-secreting cells and secreted into the mucous layer in the stomach.

It is virtually undetectable by RT-PCR in our panel of 7 gastric cancer cell lines.

0.00%

10.00%

20.00%

30.00%

40.00%

50.00%

60.00%

70.00%

80.00%

90.00%

100.00%

Normalstomach

Stomachcancer

% of casesexpressing

TFIZ1

0 %

100 %

AGS NUGC3 MKN74 KATO III SNU1 SNU5 SNU16

% of normalTFIZ1

expression

0 %

0.005 %

AG

S

NU

GC

3

MK

N7

4

KA

TO

III

SN

U 1

SN

U 5

SN

U 1

6

Wherecan we study stomach cancer?

These behaviours in patient cancer cells help determine tumour growth and the ability to metastasize.

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 divideHow easily they dieHow quickly they move

http://www.bellcoglass.com/thumbs/4506fcf.jpg

Whendo 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:

TFIZ1TFF1High expression

in oestrogen responsivebreast cancer

Binds to H. pylori

TFF1

Involved inulcer repair30% null mice get

stomach cancer

TFF1

http://www1.istockphoto.com/file_thumbview_approve/4106929/2/istockphoto_4106929_band_aid_bandage.jpg

Howare we studying this protein?

By introducing controllable TFIZ1 expression into stomach cancer cells:

pTet-Off

neoR tTA

tTAneoR

TRE TFIZ1pTRE-Tight TFIZ1

TFIZ1

TRE

TFIZ1Off

TFIZ1On

1. Transfect inregulator plasmid

2. Transfect inTFIZ1 plasmid

Stomach cancerCell line

Addingdoxycycline

turns off TFIZ1expression

The cells nowexpress TFIZ1

Whydo we study a protein in gastric cancer?

If we know

WHY the expression changesWHAT the protein does

In the normal stomachIn 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]




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.

INTRODUCTION:

Chemotherapy and radiotherapy- induced 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

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.

[email protected]

FUNDED BY: Northern Institute for Cancer Research

‘MRI scan of the breast reveals two discrete areas of abnormality, which proved to be cancer.’

http://www.breastcancer.org/pictures/diagnosis/mammogram/mri_2.jsp

http://www.ncl.ac.uk/about/




Forkhead Proteins & Cancer

Frances Purtill

iCAMB


Unregulated cell proliferation is a characteristic of all cancers

Understanding the mechanisms behind cell proliferation will help in

the treatment and prevention of cancer


M

G2 G1

S

M Phase (Mitosis)Replicated DNA is equally distributed

into 2 cells

Cell proliferation is controlled by a series of events known as the Cell Cycle

1

2

The cell cycle consists of 4 phases:

G1 and G2 are ‘gap’ phases

which prepare the cell for its

next step

3 & 4

Genetic material is copied and 2 identical cells are formed

S PhaseDNA is replicated

Forkhead Proteins & CancerForkhead Transcription Factors (FKH-TF) are critical for the regulation of the cell cycle

TARGET GENE

GENE SWITCHED ONTARGET PROTEIN MADE

FKH-TF

Forkhead proteins have been associated with various human

cancers

They bind upstream of genes involved in mitosis, and activate their expression

Forkhead Proteins & CancerForkhead proteins are vital for mitosis in all eukaryotes, from yeast to man

Yeast is easy to manipulate, genetically and biochemically

Therefore it is a valuable model to study forkhead proteins, and improve our knowledge of the

cell cycle in humans

In the future, this knowledge could potentially be used to

improve existing cancer therapies




The Development of Small Molecule

Inhibitors of the MDM2-p53 Interaction

Junfeng Liu

p53 is a Tumour Suppressor

p53+/+

p53+/-

p53-/-

1% at 18 months

% mice with tumour

74% at 6 months

2% at 9 months

Donehower et al. Nature, 1992Donehower et al. Nature, 1992Also suggest to read Donehower et al. Nature genetics, 1993Also suggest to read Donehower et al. Nature genetics, 1993

MDM2 controls p53 through an auto- regulatory negative-feedback loop

p53

MDM2

mdm2 mRNA

mRNA of other targets

p53 MDM2

p53’s proteasomal degradation

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

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

p53 p53 MDM2

MDM2

mdm2 mRNA

mRNA of other targets

p53’s proteasomal degradation

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

MDM2 protein and p53 peptide structures come from Protein Data Bank.

PyMOL Software were applied for the followed cartoons.

Structure based drug design

MDM2 Binding Domain(Binding area high lighted)


8mer p53 peptide Binding with MDM2


Three key residues of p53 in the hydrophobic pocket of MDM2


Three key residues of p53 were picked out for drug design


Drug design based on the three key residues of p53


Designed small molecule scaffold


Manual docking of the compound with MDM2 binding domain

Inhibitor Screen Using ELISA

ECL detection

DECREASED LIGHT

YY

M

Incubation of MDM2with antagonist

M M

M

M

M

ECL detection

DECREASED LIGHT

YY

M

Incubation of MDM2with antagonist

M M

M

M

M

ECL detection

LIGHT

Incubation withMDM2

M

M M

MM

YYYY Y Y

BiotinylatedIP3 Peptide

Streptavidin

M M M

YHRP conjugatedsecondary antibody

YAddition of: MDM2 primary antibody,

Cellular activity Confirmation via Western Blotting Assay

Effect of MDM2 inhibitors on the cellular levels of p53, MDM2 and p21

Nutlin-3 NU8XXX

MDM2

p53

p21

Actin

DMSO 0.5 1 10 1 10 20 M

Thank you

for

your attention!




…and tools to diagnose Lower Urinary Tract Dysfunction

having to go in the middle of the night

inability to pass any water

1 in 3 men will suffer urinary symptoms in their life:

struggling to begin peeing

having to rush to the toilet

having to go often

pain while peeing

Poor stream

‘Terminal dribble’

Incontinence

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…

…and measure the pressure to stop urine flow!

Place a small, inflatable cuff around the penis…

Pressure

Flow rate

Volume

Flow is interrupted

Pressure is measured

We can also use it to measure the bladder characteristics in healthy people to further understand the mechanics of the lower urinary tract.

Conventionally, pressure- flow measurements would require urethral catheterisation, an uncomfortable procedure with risk of infection.

Penile Cuff Machine

We can now use this non-invasive machine clinically to diagnose Bladder Outlet Obstruction in patients.

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




Heart Physiology: “The role of p50”

Silvia Gaspar PereiraPhD student with the Cell Signaling Group

Institute of Cellular Medicine

Light from the Heart Nebula - apod.nasa.gov


What does p50 do?

When the cell receives a signal, the protein p50 is released from the cytoplasm into the nucleus.

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 Aim: Is p50-dependent signaling important in signaling important in heart muscle cells?heart muscle cells?

p50

p50


To answer this question, we use mice lacking p50:

In mice without p50, hearts are significantly bigger than in normal mice.

The p50-deficient hearts are more sensitive to certain injuries, e.g. lack of oxygenation in the tissue.

p50

p50

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 If p50 is directly involved in these events, it could become a potential target for heart disease treatments!a potential target for heart disease treatments!




Regulation of the oxidative stress response in the pathogenic fungi

Candida albicans

Miranda PattersonSupervisor – Dr Janet Quinn

http://www.chuv.ch/imul/euresfun



Candida albicans

Candida albicans is the major systemic pathogenic fungi of humansIn immunocompetent people C.albicans is the

cause of common superficial infections such as oral and vaginal thrush

But, in immunocompromised people, such as people with HIV/AIDS, some cancer patients or transplant patients, C.albicans can cause life threatening systemic infections, with ≈ a

38% mortality rate

C.albicans invasion of blood vessels in the oesophagus

C.albicans invasion of a kidney from an immunocompromised rabbit

http://pathmicro.med.sc.edu/mycology/mycology-3.htm




Why are we interested in C.albicans oxidative stress responses?

Macrophages and neutrophils are cells of the innate immune system responsible for killing

micro-organisms

Macrophage

C.albicans

C.albicans cells are ingested by the

macrophage

After ingestion C.albicans starts growing hyphae

and escape the macrophage

MacrophageC.albicans hyphae

Neutrophils are important in killing C.albicans

Neutrophils kill by producing toxic reactive oxygen species in a respiratory burst

C.albicans lacking key proteins involved in oxidative stress responses are more sensitive to

killing by neutrophils and have reduced virulence

Full understanding of mechanisms used by C.albicans to overcome oxidative stresses

may reveal novel therapeutic targets!www.biology.uark.edu/dmcnabb/dmcnabb.htmlwww.biology.uark.edu/dmcnabb/dmcnabb.html

But C.albicans can kill macrophages!

http://www.biology.uark.edu/dmcnabb/dmcnabb.html

http://www.biology.uark.edu/dmcnabb/dmcnabb.html

What are we doing?

Nucleus

Cap1

Oxidative Stress

Anti-oxidant genes

Previous research shows that following oxidative stress, Cap1 translocates to the nucleus and upregulates transcription of anti – oxidant genes so that cells can survive oxidative stress.

We want to know what is regulating Cap1?

Investigating the major oxidative stress pathway in

C.albicans – the Cap1 pathway

-ive

How are we doing this?

Deleting potential key regulators of Cap1 and seeing how cells survive an oxidative

stress challenge

Control

H2O2 concentration (mM)

Wild type strainCap1 delete strainYbp1 delete strain

Wild type cells grow until high concentrations of H2O2

Cells lacking Cap1 are sensitive to H202 and can not grow even at low concentrations

Cells lacking the potential regulator of Cap1, Ybp1, are also sensitive to H202 but only at high concentrations

Have we found a regulator of Cap1 at high levels of H202 oxidative challenge?

We grow C.albicans in the presence of hydrogen peroxide (H2O2) to induce oxidative stress

Continuing research will hopefully answer this question!

Documents

PLASMA POSTER COMPETITION