Silent theatre 1 – Monday 4 November

3:35 pm-4:00 pm

Room: Hall 4


Programme session type(s): Silent theatre

Methylation Sensitive High Resolution Melting (MS-HRM) Assay for the Detection of BRCA1 and BRCA2 Promoter Hypermethylation
Speaker: Gareth Gerrard
Affiliation: Sarah Cannon Molecular Diagnostics, London, UK

Abstract:

BRCA1 & BRCA2 genes encode key components of the DNA double-strand break repair pathway. Cancers driven by loss of BRCA1/2 are associated with sensitivity to PARP inhibitors (PARPi), such as olaparib, through the synthetic-lethality of concomitantly blockading the single-strand repair pathways mediated by PARP. Monoallelic BRCA1/2 mutations require a ‘second-hit’ to the unaffected allele, since only tumours with complete abrogation of BRCA1/2 are targetable with PARPi. One recognised second-hit mechanism is gene promoter hypermethylation, which can also cause gene silencing in the absence of mutations. We sought to use extant MS-HRM protocols (based on a validated MLH1 assay) using primer and control kits from MethylDetect ApS, Denmark, to implement a BRCA1/2 promoter hypermethylation assay.

Method

14 triple-negative, grade 3 invasive ductal breast carcinoma (BC) and 13 prostate adenocarcinoma (PA) FFPE samples were sourced from the Imperial College Healthcare Tissue Bank. They were macro-dissected to obtain DNA from paired tumour and normal tissue. 100ng DNA from each was bisulphite-treated (EpiTect Fast, Qiagen) in a 20μL reaction and 3μL used in the MS-HRM reaction, along with CpG-flanking primers for either BRCA1 or BRCA2. Kit provided methylation controls were used for IQC. The MS-HRM reactions were run on a Qiagen RotorGeneQ, using EpiTect HRM mastermix (Qiagen) and analysed with the RotorGene v2.3.1.49 software.

Results

7/13 (53.8%) and 0/12 BC samples showed BRCA1 and BRCA2 promoter hypermethylation, respectively; none of the 13 PA samples were hypermethylated for either gene. 1 BRCA1 and 2 BRCA2 samples failed to yield usable results (both BC).

Conclusion

The detection of BRCA1 hypermethylation in over half of the BC samples in this limited-scale implementation of a low-cost, rapid and sensitive assay, demonstrates the potential utility of this approach for stratifying patients for PARPi therapy.

Does augmenting the complement system improve Rituximab induced cell lysis?
Speaker: Katy Plant
Affiliation: Cardiff University School of Medicine

Abstract:

Rituximab (RTX) is a therapeutic antibody used in the treatment of haematological malignancies. RTX induces complement dependant cytotoxicity (CDC). An individual’s phenotype of complement components, their “complotype”, is determined by genetic polymorphisms and governs their system functionality. We hypothesised that augmenting the complotype, by addition of purified complement components, may increase the effectiveness of RTX.



Method

Normal human serum was obtained from a healthy donor (NHS1) and from Sigma Aldrich (NHS2). The CDC elicited by NHS was assessed in Sheep Erythrocyte (ShE) haemolytic assays +/- addition of purified complement components. Studies were performed on Ramos human burkitt lymphoma cells lines. Ramos sublines, R1H (CD55/CD59 positive) and R2J (CD55/CD59 negative) were generated in house. To assess RTX induced CDC, cell viability assays were performed on Ramos cells +/- purified complement components.

Results

NHS1 induced greater CDC than NHS2. Addition of C2, C3, C5 components to NHS1 increased CDC. An estimated 2.5-fold increase of C5 concentration in NHS1 increased ShE lysis by 20.2%. An estimated 10-fold increase in the C2 and C3 concentrations in NHS1 increased ShE lysis by 39.7% and 21.8% respectively. Addition of the other C-Components did not augment CDC on ShEs.

R1H cells are resistant to RTX induced CDC. R2J cell viability post RTX sensitization was 23.8%. Addition of C2, C3 and C5 decreased cell viability to 9.7%, 6.5% and 11% respectively. The other C-Components did not augment RTX induced CDC.

Conclusion

Our data implicate C2, C3 and C5 as limiting factors of CDC and suggests addition of these to RTX therapy could improve efficacy. This is the first preliminary data exploring therapeutic antibodies and complement addition. Research into this hypothesis, the theoretical clinical potential and safety profile is warranted. The next step in developing knowledge in this area would be further R1H studies to prove utility in refractory cancer cells.  

The NIHR STAMINA (Supported exercise TrAining for Men wIth prostate caNcer on Androgen deprivation therapy programme): data from year one.
Speaker: Sophie Reale
Affiliation: Sheffield Hallam University

Abstract:

There are approximately 250,000 to 300,000 men in the UK on androgen deprivation therapy (ADT) for prostate cancer (PCa). ADT carries well-documented substantial, chronic adverse effects on quality of life including intractable fatigue, increased CVD risk, suicidal intent, sexual dysfunction and reduced muscle function. Supervised exercise therapy is the only evidence-based treatment that improves cancer-specific quality of life, highlighted in national and international best practice guidelines. In the ‘real world’, this is not offered to these men. The NIHR-funded STAMINA programme is an international ‘first-in-class’ study, testing embedding such treatment in core cancer services nationally. We report findings from intervention development and pre-pilot work streams.

Method

A national survey describing cancer care pathways and exercise services in prostate cancer (for intervention tailoring and describing usual care), interviews and focus groups with stakeholders: NHS health care professionals (HCPs), community-based exercise professionals and pre-pilot data from men on ADT at three NHS trusts.  

Results

Data from 78 NHS trusts revealed complex and diverse PCa management pathways. Complexity derives, inter alia, from the split between urology and oncology for diagnostic, referral, treatment and follow-up pathways – according to entrenched cultures and behaviour. Data from one-to-one interviews with HCPs (n=35) and focus groups with exercise professionals (n=4, total n=22) highlighted that staff training needs, capacity issues and time pressures will need to be addressed if we are to successfully design and evaluate our new embedded care pathway. However, if this can be done, NHS HCPs and community exercise partners are prepared to participate in our planned prospective evaluation of long-term intervention effects in a cluster RCT in men on ADT involving 40 NHS trusts and over 1000 men on ADT from mid-2020. In our pre-pilot cohort study 28 men on ADT (to date) have been referred from routine NHS cancer clinics to exercise programmes individually tailored according to functional capabilities and comorbidity profile, delivering adequate dose without compromising safety.

Conclusion

Data from preparatory work streams has provided critical insight into how innovative care models can be developed and rigorously trialled so the goal of cost-effective, clinically-relevant improvements in cancer specific quality of life can be attained.

The Trigger project: Introducing electronic patient reported outcome measures into radiotherapy services
Speaker: Amy Sharkey
Affiliation: Macmillan Cancer Support, London, UK & Royal College of Radiologists, London, UK

Abstract:

Patients receiving pelvic radiotherapy can experience long term gastrointestinal side effects post-radiotherapy. The Trigger project identifies patients experiencing symptoms of radiation-related bowel toxicity using the ALERT-B questionnaire, and directs them to the appropriate clinician.

Trigger is a service evaluation project, aiming to prove the utility of electronic patient reported outcome measures (PROMs), and to demonstrate the feasibility of a low-resource project as a model for collecting PROMs. It is a collaboration between Macmillan Cancer Support, the Royal College of Radiologists, and three NHS Trusts: Velindre, Imperial College Healthcare and Brighton and Sussex University Hospitals.

Method

Patients register on the Trigger website, hosted by My Clinical Outcomes, and receive periodic emails to complete the short ALERT-B questionnaire electronically, to screen for long-term bowel symptoms which could have been caused by pelvic radiotherapy. If answering “yes” to any of the questions, patients are directed to appropriate services.

 

Six months following the completion of their radiotherapy, patients are sent a separate questionnaire to evaluate the utility of the project.

 

Results

336 patients registered in first the 9 months across the 3 sites. Patients with a range of different cancers signed up: anal (2%), bladder (1%), prostate (87%), rectal (4%) and gynaecological (6%).

 

43 patients (of 65 eligible (uptake 66%)) have answered their 6-month post treatment questionnaire thus far, and 72% answered “yes” to at least one of the ALERT-B questions. 85% of responding patients reported they found the Trigger project helpful.

Conclusion

These promising results show that electronic PROMS can be introduced in radiotherapy departments using a low resource model. The Trigger project works as a feasibility model, showing patients engage with electronic PROMs projects, and find them useful. PROMs for other tumour types could be collected in a similar manner, based on the low-resource model used here, using site-specific PROMs based on the ALERT-B tool.