1. Biomarker Screening Pilot

2. Overview Background and project aims
Required site-set up for participation
Approaching patients to participate in biomarker- screening
Pilot lab kit
Tumour block Collection logistics
Blood collection logistics
Saliva collection logistics
Sample Queries
Restocking kits

3. Background and project aims
Establish processes that enable retrieval and analysis of samples (saliva, blood and FFPE tumour tissue) within specified timelines
Establish pathology contacts in all sites where diagnostic tissue blocks may be stored.
Evaluate the feasibility of undertaking DNA/RNA sequencing using each sample type to inform the future approach to biomarker-screening

4. Rucaparib
Arm M of the study will contain Rucaparib which is a PARP inhibitor.
Patients will be defined as biomarker positive if a known relevant mutation is detected in a gene involved in DNA repair, this includes BRCA1, BRCA2, ATM and 11 other genes
Only patients that are biomarker positive will have the chance to be randomised into the Rucaparib arm.
All biomarker negative patients can enter the other arms of the study.
All suitable patients looking to enter the STAMPEDE study will need to complete biomarker analysis once the arm is greenlit at that site.
Sites that take part in the pilot will be accredited to start the new arm in the spring.

5. Required site-set up for participation
A variation to the clinical trials agreement (CTA) was put in place for the retrospective tumour block project (Variation Number 3). 
All sites should have CTA variation 3 in place.
Most sites will not need new variation to the CTA as variation 3 states additional funding will be made available.
A signed STAMPEDE Histopathology Agreement will be required from all sites before being accredited to participate in the trial.

6. Approaching patients
To be eligible to participate in the biomarker-screening pilot, patients are required to provide informed consent to both:
Main STAMPEDE study consent
Additional Research consent
The following selection criteria must be met:
Confirmed metastatic disease (M1)
Histologically confirmed prostate adenocarcinoma with recent tumour rich FFPE blocks (obtained within 8 months of registration)
If hormone therapy has started, FFPE tumour blocks should be sent within:
8 weeks of the patient starting LHRH.
10 weeks of starting anti-androgens.

7. Registration
Screening required prior to registration: Radiologically confirmed metastatic disease and minimum baseline bloods are required.
Once eligibility for the biomarker screening pilot has been confirmed, the patient must be registered in order to proceed to biomarker screening.
Once patient consent has been obtained Form 01A Registration CRF should be completed.
The patient can be registered for biomarker screening by phoning the MRC CTU at UCL on +44 (0) While making sure to have the registration CRF to hand.
A registration ID will be provided by the STAMPEDE trial team.
The registration ID should be used on all data collection forms and correspondence with the MRC CTU at UCL prior to randomisation.

8. S-Stampede screening flowchart

9. S-Stampede screening flowchart

10. Pilot lab kit Materials provide for each sample: Tissue sample
1 x Registration ID label
1 x Plastic envelopes per sample
1 x Padded envelope (pre-paid postage)
1 x Test Request Form
Blood sample
1x Transport box
2 x 10ml blood collection tubes (cell free DNA StreckTM tubes)
2 x Postal tubes
1 x Addressed padded envelope (pre-paid postage)
1 x S-STAMPEDE Blood CRF
Saliva sample
1 x Saliva collection kit (Oragene DNA)
2 x Sample barcode
1 x Plastic envelope
1 x Sample Requisition form
1 x padded envelope (pre-paid postage)

11. Biomarker screening sample collections
Sample type
Registration
Prior to starting research treatment
Pre-progression (48, 72 & 84 weeks)
Progression
Post progression, pre-2nd line treatment
2 tumour rich FFPE tumour blocks obtained within 8 months of registration
X #
Blood
(StreckTM tubes) X
n/a
Saliva X*
# Two tumour rich FFPE blocks should be sent for testing (Almac Diagnostic) immediately following registration within the required hormone therapy timelines
* Saliva sample using the Oragene 510 kit should be taken after registration

12. FFPE block analysis Key points
Please request the two most tumour rich FFPE tumour blocks obtained within 8 months prior to registration.
Please also complete the Test Request Form and send with the tumour samples when shipping to the central lab.
Almac diagnostics will contact sites regarding queries
Biomarker results are not required to randomise patients until the “rucaparib comparison” is active at your site
Funding for reimbursement per participant is available. A £60 payment will be made available on receipt of a tumour rich sample. An additional sum of £40 will be paid following successful DNA/RNA extraction and sequencing completed.

13. FFPE blocks - Processing and shipping
Place both FFPE tumour blocks belonging to one patient into one zip lock bag.
Complete a sample label and attach to each zip lock bag using the stickers provided.
Complete a Test Request Form and make 2 copies, send one with the samples, and retain one on-site.
Send to the Biomarker test provider, Almac Diagnostics using the pre-paid envelope provided.
Please ensure that the postage envelopes are securely fastened using tape or staples to ensure samples cannot fall out in transit.

14. Blood sampling Similar to current sub-study blood sampling practise
Blood collection using cell free DNA StreckTM tubes from which circulating tumour DNA can be extracted

15. Blood sampling - Processing and shipping
Complete label on blood collection tubes, add the registration ID
Collect 2x 10ml blood using StreckTM tubes
Invert 8 to 10 times
No onsite processing required
Store at room temperature
Complete S-STAMPEDE CRF
Post samples and CRFs – within a maximum of 5 days to the ICR

16. Saliva sampling
Color Genomics will supply Oragene 510 saliva kits for the Biomarker-screening pilot study. These kits are different to the Oragene 500 saliva kits.

17. Saliva sampling- Processing and shipping
Adhere one of the barcode labels onto the Sample Requisition Form.
Complete sample ID label and stick to the sample tube avoiding covering the barcode.
Place the sample into the sealable plastic bag and put into the original packaging.
Once completed seal the kit into the pre-paid envelope provided.
Barcodes

18. Queries - Almac diagnostics
Almac will direct queries at site by and phone.
The CTU will occasionally contact sites regarding escalated queries
Examples of queries
Registration number on documents does not match sample labelling.
Documentation not correctly completed

19. Restocking kits 2 lab kits will be sent to sites.
The CTU will send out a new kit once Almac have confirmed a site has sent a tissue sample for analysis.
If a site believes they may see more then 2 suitable patients at one time please contact the STAMPEDE inbox as soon as possible.

20. Questions?