DEVELOPING

THERAPEUTIC
STRATEGIES FOR MPM
CANCER WITH LOSS OF
BAP1 USING NEW
PRECLINICAL MODELS

Mohammad Reza Zare

Mres Biomedical Science and Translational Medicine
University of Liverpool
Backgrounds

BAP1 inactivation in MPM

• Mesothelioma characterized into 3 types: Pleural Mesothelioma (80%),

Peritoneal mesothelioma (20%) and Pericardium (<1%)

• Mesothelioma histological classifications:

Epithelioid, Biphasic, Sarcomatoid and Desmoplastic

• Malignant Pleural Mesothelioma (MPM) has aggressive malignancy

• Ca u s e d b y e xp o s u r e t o a s b e s t o s

• Latency period of ~40 years

• Ri g h t s i d e d p l e u r a l e f f u s i o n , t h i c k e n i n g a n d l o s s o f l u n g v o l u m e CT scan shows the extensive pleural

thickening that is characteristic of
mesothelioma, effusion, and
• M P M C au s e s r e d u c t i o n i n e x p r e s s i o n o f m o l e c u l e s i n p 5 3 p a t h w a y reduction in the volume of the
affected hemithorax
• M P M c a u s e s l o s s o f B RC A - a s s o c i a t e d p r o t e i n 1 ( B A P 1 )
(Taken from Medscape Tan et al
2021)
Chorioallantoic membrane (CAM)

• Highly Vascularized

• Grows around 7 days

• Self sufficient in terms of nutrition

• Easily accessible in/ ex ovo

• Cost effective technique

• Easy administration of drugs

• Animal replacement techniques (Xenograft

(Taken from Lui et al 2013)
models)
 Chick Embryo CAM’s Short experimental Timeline

Incubation
Start E0
Day 3 Day 7 Day 14

Windowing Eggs Implanting Tumour Dissection and Imaging

Cell lines
Tumour
Project Timeline

RRM1/RRM2 • Cell Culture • Tu m o u r • RNA • qPCR

Primer design ( M STO - 2 1 1 H dissection (Expression
Extraction
Cell line) of RRM1
• Seeding of isogenic
• CAM formation and RRM2
RNA dishes BRs and
Tr a n s c r i p t s )
• Tu m o u r tumours
Implantation

00 01 02 03 04 05 06 07 08 09 10 11

Start
Results

Validating Primers
X3A
• Endpoint PCR to validate
the designed Primers
• Contamination in RRM1

X3C
• No contamination in RRM1
Results

Primer Effi ciency

% efficency 96.78 % efficency 101.66 % efficency 89.93

MSTO-211H Cell line

• Human Cell line ( Homo sapiens)

• Established from pleural eff usion of a patient with biphasic mesothelioma

• High binding affi nity for EGF

• Express Neuron specifi c enolase (NSE) and ⍺ /β human chorionic

gonadotropin (HCG)

• Splitted Cells ever y two days (80-90% Confl uency)

Experiment 4 Dissection Imaging
Experiment 5 Dissection Imaging
Results

RNA Analysis

• Two Strong clear bands (28S and 8S RNA)

• High MW bands illustrate contaminating

genomic DNA

• Smearing of RNA bands towards lower

MW products Could be sign of RNA
degradation

Hyperladder BR1 BR2 BR3 chB T. chC T. chD T. chA T. chB T. chF T. Hyperladder
qPCR data RRM1
qPCR data RRM2
 

Future Plans

• Expression of RRM1 and RRM2

Transcripts in 3D

• Use of CAM model in vivo for drugs

testing

Thank you for your

Attention!