BriTROC personalised biomarkers in relapsed ovarian high grade - - PowerPoint PPT Presentation

Iain McNeish Professor of Gynaecological Oncology Wolfson Wohl Cancer Research Centre Institute of Cancer Sciences University of Glasgow, UK

BriTROC – personalised biomarkers in relapsed

• varian 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

6 12 18 24 30 36 42 48 1 10 100 1000 10000

Time (months) CA125 (units)

Surgery + Carbo Carbo Taxol Gem Carbo Cis/etop V.E. 61 retired school teacher

January 1998

6 12 18 24 30 36 1 10 100 1000 10000

Time (months) CA125 (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 study

Carboplatin-based chemotherapy

1 6 12 18 months

100 biopsies platinum-resistant 200 biopsies platinum-sensitive Sample 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

NBF Histoscore Histoscore r = 0.846 50 100 150 200 50 100 150 200 UMFIX NBF 50 100 150 200 UMFIX histoscore r=0.844

NBF Histoscore

UMFIX histoscore UMFIX NBF 50 100 150 200 Histoscore 50 100 150 200 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

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 T

• xicity
• 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 results

Relapse sample Diagnostic sample

BriTROC – some quick results

Patient 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

Acknowledgements

University of Glasgow Darren Ennis Suzanne Dowson Josephine Walton Malcolm Farquharson Elaine Leung Oliver Hofmann University of Cambridge James Brenton Anna Piskorz T eodora 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