Iain McNeish

Professor of Gynaecological Oncology

Wolfson Wohl Cancer Research Centre

Institute of Cancer Sciences

University of Glasgow, UK

BriTROC – personalised biomarkers in relapsed

ovarian high grade serous carcinoma

GCIG Translational Committee 4th June 2016

Big translational questions in HGSOC

• Can we develop novel therapies for HGSOC with accurate predictive biomarkers? (Clinical)

• How does recurrent HGSC evolve into platinum-resistant disease? (Translational)

• Can we develop decent pre-clinical models of HGSC? (Basic science)

HGSC in two patients

0 6 12 18 24 30 36 42 481

10

100

1000

10000

Time (months)

CA

125 (

units)

Surgery +

Carbo

Carbo

Taxol

Gem

CarboCis/etop

V.E. 61 retired school teacher

January 1998

0 6 12 18 24 30 361

10

100

1000

10000

Time (months)

CA

125 (

units)

CarboTaxol

With IDS

Carbo

Caelyx

Carbo w.Taxol

M.W. 63 retired school teacher (BRCA2VUS)

January 2013

Why does HGSC stop responding to platinum?

Refractory Resistant Partially sensitive Sensitive

Carboplatin-based

chemotherapy

1 6 12 18 months

0% 10% 30% 60%

Probability of responding to platinum at relapse

Data from stage IIIc/IV HGSOC patients in Scotroc1 trial (Vasey et al JNCI (2004) 96:1682)

CCNE1, MYC BRCA1, BRCA2

Potential

biological

drivers

BriTROC collaborative

• UK Translational Research in Ovarian Cancer Collaborative

• 12 major UK ovarian cancer centres (Cambridge, Glasgow, Newcastle, Imperial,

Barts, Edinburgh, Leeds, Manchester, Belfast, Birmingham, Bristol, Kings)

• Ovarian Cancer Action provides infrastructure funding

James Brenton

BriTROC1 Study

Sample Collection Study to Investigate the Role of

Homologous Recombination Deficiency in

Platinum Sensitivity in Recurrent High Grade

Serous Ovarian Cancer

BriTROC1 study

• Multi-centre, study.

• Study recruitment: 300 biopsies/samples (c.12 sites in the UK) over 4 years.

• Timelines:

- Open to recruitment 14th December 2012

- Planned accrual completion December 2016

BriTROC studyCarboplatin-based

chemotherapy

1 6 12 18 months

100 biopsies

platinum-resistant

200 biopsies

platinum-sensitiveSample from

diagnosis

Q2

Q1

Q3

Refractory Resistant Partially sensitive Sensitive

Study Objectives

• Safety and feasibility of obtaining recurrent biopsies.

• To obtain 300 biopsies from women with relapsed HGSC.

• To assess Homologous Recombination (HR) genes in relapsed HGSC

compared with the same genes in tumours at first presentation.

• To compare HR genes in platinum-sensitive vs platinum-resistant

relapsed HGSC.

• To compared somatic mutations with those identified in ctDNA

Inclusion Criteria

• Relapsed HGSC (+ G3 endometrioid)

• At least one line of platinum-containing chemotherapy

• FFPE material from time of diagnosis available (NOT ascites

cytology).

• Disease suitable for biopsy

• Patients also allowed to enter at secondary debulking

Exclusion Criteria

• Non-HGSC pathology

• Original diagnosis of high grade serous cancer made on

cytology only

• Disease not amenable to biopsy

Samples and Sample Collection

• 2 x 14–16G cores – fixed in methanol (UMFix)

• Access to ≥1 archival FFPE block.

• 20ml blood for plasma for ctDNA.

• Extra 20ml for genomic DNA extraction

• Optional 10ml blood immediately prior to both first and

second cycles of chemotherapy following biopsy for ctDNA

Why UMFix? - IHC

Formalin UMFix

CK7

p53

NB

F H

isto

sco

re

His

tosc

ore

r = 0.846

0 50 100 150 200

0

50

100

150

200

UMFIX NBF

0

50

100

150

200

UMFIX histoscore

r=0.844NB

F H

isto

score

UMFIX histoscoreUMFIX NBF

0

50

100

150

200

His

tosc

ore

0 50 100 150 200

0

50

100

150

200

Piskorz et al (2016) Ann. Oncol. 27:532

Why UMFix? - DNA

Piskorz et al (2016) Ann. Oncol. 27:532

Greater DNA yields

than formalin

Better copy number

estimation than

formalin with less noise

BriTROC Recruitment

N = 207 (26th May 2016)

150 = platinum-sensitive

43 = platinum-resistant

BriTROC Recruitment

0 24 48 72 96 120 144 168 192

Months since diagnosis

1

1

2

No. prior lines chemo

Platinum-resistant relapse

(median 2 prior lines)

Platinum-sensitive relapse

(median 1 prior line)

2

5

3

7

Screening and toxicity

Screening after 150 recruited

• 322 patients screened

• 67 declined

26 declined biopsy

20 needed to start chemotherapy urgently

2 too much pain

19 others

• 120 ineligible including

60 due to disease not accessible for biopsy

17 contra-indication to biopsy

20 due to no FFPE material from diagnosis

Toxicity

• 3 biopsy-related SAE

2 = grade 2 pain

1 = grade 2 haemorrhage (liver)

DNA yields

Biopsy median = 2.76 µg (range 0.05 – 18.7)

Surgical median 6.73 µg (range 0.52 – 65)

Number of failures = 17/114 (15%)

14 biopsy, 3 surgical

90/114 (79%) samples yield >200ng DNA

Personalised biomarkers of response in ovarian

high-grade serous cancer

James D. Brenton

Iain A. McNeish

The science programme

Aim 1: How does relapsed HGSC differ from time of diagnosis?

1. Sequencing of BriTROC relapse samples

TAm-Seq – cancer panel at great depth (2000x)

Shallow whole genome sequencing - CNA

Deep whole genome sequencing - SV

Comparison with sample at time of diagnosis

Germline DNA sequencing

2. Clonal heterogeneity

Detailed re-sequencing of primary tumour

BriTROC – some quick resultsRelapse sample

Diagnostic sample

BriTROC – some quick resultsPatient number 1. Diagnosed 20 months prior to enrolment. Two prior lines of chemo, platinum-resistant

relapse

TAmSeq: (archival and relapse) TP53 H179Y PIK3CA E542K

BriTROC – some quick results

Patient number 17. Diagnosed 30 months prior to enrolment. One prior line of chemo, platinum-sensitive relapse

TAmSeq (archival and relapse): TP53 R282W

Conclusions part 2 and future direction

• Obtaining tissue in relapsed HGSC is feasible and safe

• p53 mutations are constant

• Deep WGS on relapsed samples

• Define utility of ctDNA

AcknowledgementsUniversity of Glasgow

Darren Ennis

Suzanne Dowson

Josephine Walton

Malcolm Farquharson

Elaine Leung

Oliver Hofmann

University of Cambridge

James Brenton

Anna Piskorz

Teodora Goranova

Anna Supernat

Geoff McIntyre

CRUK Clinical Trials Unit Glasgow

Liz-Anne Lewsley

Diann Taggert

Jim Paul

Beatson West of Scotland Cancer Centre

Ros Glasspool

David Kay

BriTROC investigators

Hani Gabra

Charlie Gourley

Andrew Clamp

Michelle Lockley

Geoff Hall

Richard Kennedy

Sudha Sundar

Axel Walther

Ana Montes

Marcia Hall

CRUK Beatson Institute

Karen Vousden

Julianna Blagih

David Stevenson

Karen Blyth