The Cell Cycle Cancer Test the Future of Personalised Healthcare 2 CONFIDENTIAL G2 G1 M S ORC Cdc45 Cdc6 MCM Replisome Geminin Cdt1 Cdc7 CDK ASK Cyclin ... CONFIDENTIAL Three Ways

www.fglabs.com CONFIDENTIAL

The Cell Cycle Cancer Test

1

Shaping the Future of Personalised Healthcare


www.fglabs.com CONFIDENTIAL 2

G2 G1

M

S

Cdc45 Cdc6 MCM Replisome

Cdt1 Geminin

Cdc7 CDK

ASK Cyclin

P P

ORC

License to Replicate

The DNA Replication Initiation Machinery

Cell Cycle


Three Ways Out of Cycle

quiescence

senescence

differentiation

Cdc6

ORC MCM



DNA replication

initiation proteins in

tissue maintenance and cancer

Introduction



DNA replication

initiation proteins in

tissue maintenance and cancer

From bench

to bedside – new tests

for cancer detection and prognosis

Introduction



From bench

to bedside – new tests

for cancer detection and prognosis



Requirement for Prognostic and

Predictive Markers in Breast cancer

50% of patients who

receive loco-regional treatment

will never relapse, whereas

the remainder will die of

metastatic disease





50% of patients who

receive loco-regional treatment

will never relapse, whereas

the remainder will die of

metastatic disease

50%





1/3 of patients who are node-

negative will experience relapse

1/3





1/3 of node-positive patients not

receiving adjuvant therapy will be

disease-free after 10 years

1/3

1/3



Cell Cycle Phase Progression

in Breast Cancer

G2 G1

M

S

H3

MCM

P

Geminin

MCM2 Ki67 Geminin

Aurora A




Tumour biopsy material

Multiparameter analysis:

Mcm2, geminin and H3S10ph

Mcm2 high (>30%)? No

No

Geminin or

H3S10ph high?

Good Prognosis

Phenotype I







No No

Yes

Geminin or

H3S10ph high? Geminin or

H3S10ph high?

Good Prognosis Good Prognosis

Phenotype II







No No

Yes

Geminin or

H3S10ph high? Geminin or

H3S10ph high?

Good Prognosis Good Prognosis

Yes

Poor Prognosis

Phenotype III


Cell Cycle State Predicts the Outcome

of Breast Cancer after Surgery

Days from diagnosis to death, recurrence or last follow up

Pro

po

rtio

n r

em

ain

ing

fre

e f

rom

recu

rren

ce





Clone generation

Clonal stability and Isotype homogeneity

12

Accelerated antibody stability study

7

0

0.5

1

1.5

2

2.5

3

TP0 TP1 TP2 TP3 TP4

Intensity scores of 37°C stability study

Negative

1 1/10

2 1/10

3 1/10

4 1/10

5 1/10

6 1/10

7 1/10

Immunisation

Clone pre-selection (ELISA)

many clones

3

Development

IHC titration and reproducibility

11



IHC titration curve Mcm2 antibodies

Mcm2 BD Ab



Validation in tissues with varying expression levels

Placenta Colon Fallopian tube

High Medium Low



Stability testing of Ab clones

Select stable clones at a minimum of 18 months equivalent real time

37°C Stability Study day 80


Over and under treatment for patients

with breast cancer?



Over and undertreatment for patients

with breast cancer?

Chemotherapy No Chemotherapy

29% (n=9)

71% (n=22)

Phenotype I

24% (18/76) of patients

received adjuvant

chemotherapy with

minimal benefit



Over and undertreatment for patients

with breast cancer?


20% (n=9)

80% (n=36)

Phenotype II


received adjuvant

chemotherapy with

minimal benefit



Over and under treatment for patients

with breast cancer?


54% (n=57)

46% (n=49)

Phenotype III


did not receive treatment

with adjuvant

chemotherapy



Failed trials? Let FGL find out why…

Plasma Membrane

Receptor Protein

Growth Factor

Signal

Transduction Cascade

Out-of-cycle Phenotype I

Cell Cycle



Failed trials? Let FGL find out why…

Plasma Membrane

Receptor Protein

Growth Factor

Signal


G1 Arrest Phenotype II

G1



Failed Clinical trials? Let FGL find out why…

Plasma Membrane

Receptor Protein

Growth Factor

Signal


Actively Cycling Phenotype III

G2 G1

M

S

G1




Advantages

• Can be used for all forms of breast cancer

• Simple, inexpensive IHC based test

• Identification of CT sensitive and insensitive tumours for breast cancers

• Enables patient stratification for clinical oncology trials

• Applicable to FFPE samples

Thank You www.fglabs.com

Documents

The Cell Cycle Cancer Test the Future of Personalised Healthcare 2 CONFIDENTIAL G2 G1 M S ORC Cdc45 Cdc6 MCM Replisome Geminin Cdt1 Cdc7 CDK ASK Cyclin ... CONFIDENTIAL Three Ways