Greg BrookeMolecular Oncology

Prostate Cancer- Leading Cause of Cancer-related death in men- Growth dependent on androgens- Therapy targets androgen action- Therapy failure is a significant clinical problem

Breast Cancer- Leading Cause of Cancer-related death in women- Growth often dependent upon oestrogens- Therapy targets oestrogen action- Therapy failure is a significant clinical problem

Prostate Cancer growth is dependent upon the Androgen Receptor Pathway

The Androgen Receptor Pathway

ARE ARE

HSP complex

Corepressors

DHT Antiandrogen

The AR Pathway Androgen Blockade

Coactivators

GROWTH GROWTH

Potential Biomarkers/Therapeutic

Targets

Characterising proteins that regulate AR action

- Inhibition of critical interactions

Mechanisms of therapy resistance

FUS is an Important Regulator of Prostate Cancer Growth

0 8 16 24 48 72Et

OH

MIB

Time (hrs)

FUS-actin

FUS-actin

blot

0

1

2

3

Time (days)

4

5

6

7

0 2 4 6 8

Rel

ative

Gro

wth

*

***

***

***LNCaP FUS

- DOX + EtOH+ DOX +EtOH- DOX + MIB+ DOX + MIB

0.5

0.7

0.9

1.1

1.3

1.5

1.7

0 5 10 15 20 25 30 35 40

Time (days)

Tum

ou

r si

ze

Increasing FUS

Decreasing FUS

Increasing FUS

IN VIVO

Brooke et al. 2011. Cancer Research. 71; 914-924

ER ER

ERE ERE

E2 Antiestrogen

AR AR

ER ERCorepressors

The ERα Pathway Hormone therapy

Coactivators

GROWTH GROWTH

The Androgen Receptor Pathway Can Drive Therapy Resistance in Breast Cancer

GROWTH

1 1.5 2 2.5 3 3.5 4 4.5 50.7

1.1

1.5

1.9

2.3

2.7

E2 (M)

Rela

tive

prol

ifera

tion

1 1.5 2 2.5 3 3.5 4 4.5 50.7

1

1.3

1.6

1.9

MiB (M)

MCF7TAMRLTED

E2 (nM) MIB (nM)

*** *** *** ***** ***

**

0

100101

0.1 0

100101

0.1

Androgens stimulate the growth of endocrine resistant breast cancer

Inhibition of AR may be a valid method to inhibit endocrine resistance

Confocal imaging and FRET (with Phillippe Laissue)

RNA-seq and ChIP-seq

Characterisation of AR/ERα crosstalk

AF-1

LBD LBD

AF-1

AR1-54 Repressor Motif

ARE ARE

AF-1

LBD

Coactivators Corepressors

Engineered Repressors are potent inhibitors of Androgen Receptor Activity

Brooke et al. Submitted

[AR1-54] ng [MAD7-35AR1-54] ng

Isolated domainFused domains

Repression domain /interaction motif:

MockREP2-IM

**

**

0 1 2 3 4 5 6 70

5

10

15

20

25

30

35

Time (days)

Rela

tive

cell

num

ber

Inhibition of AR activity

% a

ctivi

ty

Repressor

Inhibition of cell growth

Techniques and Collaborations• In cell and in vivo models (Honorary Contract with Imperial)

– Transcription assays, 2-hybrid assays, confocal, ChIP, growth assays, FACS, DOX-inducible cell lines, motility assays……

• Collaborations– Common interests in Breast and Prostate Cancer e.g. FUS and CTCF/BORIS (Elena)– Peptide inhibitors of AR (Jody)– Fluorescent Microscopy – AR/ER in BCa (Phillipe)– Proteomics (Metodi)

• Future Collaborations?– Protein aggregations

• FUS (Amyotrophic Lateral Sclerosis)• Androgen Receptor (Kennedy’s Disease)

– Metabolomics