1. The UK ME/CFS Biobank Improving health worldwide www.lshtm.ac.uk Dr. Eliana Lacerda Ms. Erinna Bowman

2. Presentation overview Some background information What PWME have told us about tissue donation The UK ME/CFS Biobank target population Recruitment strategy & procedures Processing of samples Resources available Procedures for accessing the samples

3. Some background information Myalgic Encephalopamyelitis/Chronic Fatigue Syndrome (ME/CFS) long-term disabling medical condition, usually presenting with severe and overwhelming fatigue after mental or physical activity accompanied by a range of disabling symptoms such as cognitive impairment, sleep problems, pain, autonomic dysfunction, which follow abnormalities in the nervous and immunological systems

4. ME/CFS may affect at least 120,000 in England is almost 4 times more common in women Prevalence is inversely related to specificity of diagnostic criteria Cases are seen in individuals beyond young adulthood Some background information

5. Action for ME The ME Association ME Research UK Private donor 2011 – The UK ME/CFS Biobank Project set up at the LSHTM Jointly funded by:

6. What PWME have told us about tissue donation for research Generally a positive response: “happy to donate” Reasons – Altruism – Taking part “in a new exiting step forward” – Empowerment ‘’also you feel sort of powerless and so much is taken away from your life. I think that it might have a good psychological impact if people felt like they were able to do something positive. Because it made me feel, just coming here today that I might be doing a little bit…”

7. What PWME have told us about tissue donation for research Ensuring quality and linking to relevant data “and if it is done, like really thoroughly, so the samples are going to be: i) very good quality, and ii) have the appropriate information attached, and because we don’t know what area it’s going to go down, we have to be quite broad in the information, like basic stuff like age, length of illness need to be there at onset, as much extra information as possible…”

8. What PWME have told us about tissue donation for research Some concerns “if you’re trying to get as much people as you can, they are afraid of what you’re going to do, whether the government would get a hold of it, whether the insurance companies could use it, or whether benefit agencies would use it”

9. The UK ME/CFS Biobank target population Cases: confirmed ME/CFS (Canadian or CDC-94 criteria) – Age 18-60 years – Consent Controls: ‘healthy’ – Frequency matched by age, gender and area of residence

10. Recruitment strategy NHS primary and secondary care services – Norfolk and Suffolk ME/CFS Specialist Service – Primary Care Research Network (PCRN) – East of England – Collaboration with other NHS Services Department of Clinical Immunology of the Royal Free Hospital (Dr. Chee) ME/CFS Disease Register

11. Our recruitment, so far... Recruitment Site CasesControls Male (n) Female (n) Total (n) Male (n) Female (n) Total (n) ME/CFS Disease Register11 134 Norfolk and Suffolk ME/CFS Specialist Service64349224 East of England PCRN Total264975111223 RFH – Clinical Immunology Department34711 Total35107142141832 Consenting controls Consenting cases

12. Recruitment strategy Norfolk & Suffolk ME/CFS Specialist Service Royal Free Hospital East of England PCRN

13. Recruitment strategy Ethics IRAS - Integrated Research Application System – NHS NRES/Research Ethics Committee (REC) London-Bloomsbury (11/LO/1760) – NHS / R&D offices (CSP 77765) East Coast Community Health Care CIC NHS Norfolk NHS Great Yarmouth & Waveney NHS Suffolk Joint Research Office (Royal Free site) LSHTM Ethics Committee approval HTA license Informed consent & free withdrawal All samples anonymous

14. Outline of recruitment procedures Possible cases: NHS ME/CFS Service or ME/CFS DR members Possible controls - invited by consenting PWME Collection of samples blood (+ urine) Processing and storage of blood samples 1. Invitation of participants 2. Recruitment of consenting subjects Clinical assessment Exclusion 3. Sample collection 4. Confirmation of diagnosis, sample processing and storage Lab tests results(NHS) Confirmed ME/CFS cases Non- ME/CFS chronic fatigued group Consent given + criteria met Probable case or control Controls UCL/Royal Free Biobank

15. Recruitment procedures Assessment of participants Symptoms assessment – self-reported Clinical measurements – health professionals Extended questionnaires – self-reported Laboratory tests

16. East of England PCRN Hub-and-spoke model Rosedale Surgery Andaman Surgery Alexandra & Crestview Surgeries Beccles Medical Centre Bridge Road Surgery Victoria Rd Surgery Recruitment procedures

17. Processing of samples (within 6 hours of collection) 1 x 5ml Clotted Vacutainer for Serum Upon blood draw invert the tube 180º 5-6 times Allow sample to clot fully (minimum recommended time -30 min) Centrifuge at 1300-2000g for 10 min at room temperature (18-25°C) 5 x 10ml Sodium Heparin Vacutainers Centrifuge at ≤1300g for 10 minutes at room temperature (18-25°C) Remove 7x250ul aliquots of plasma Replace plasma volume with medium Separate PBMCs from granulocytes/ RBCs using BBSOP 007v1 Remove RBC pellet into 1x2ml aliquot for storage in vapour phase liquid nitrogen Cryopreserve PBMCs into 17x1ml aliquots at 5 million cells/ml for storage in vapour phase liquid nitrogen. Prepare 10 x 250ul aliquots for storage in vapour phase liquid nitrogen Upon blood draw invert the tube 180º 5-6 times 1 x 10ml K2-EDTA Vacutainer Upon blood draw invert the tube 180º 8-10 times Centrifuge at ≤1300g for 10 minutes at room temperature (18-25°C) Prepare 2x500ul aliquots of whole blood for storage in vapour phase liquid nitrogen Prepare 3x1ml aliquots of plasma for storage in vapour phase liquid nitrogen Separate PBMCs from granulocytes/ RBCs using BBSOP 007v1 Cryopreserve PBMCs into 3x1ml aliquots at 5 million cells/ml for storage in vapour phase liquid nitrogen PAXgene Vacutainer Store the tube at room temperature (18-25°C) for 2-72 h Upon blood draw invert the tube 180º 8-10 times Freeze the tube upright in a wire rack at -20°C for 24 h Transfer the tube to -80°C for storage

18. Resources available Blood products stored (per participant) Whole blood Serum Plasma Red Blood Cells (RBCs) Peripheral Blood Mononuclear Cells (PBMCs) Preparations for RNA extraction (PAXgene vacutainer)

19. Resources available Baseline clinical data, including: – Clinical measurements » blood pressure (seated and standing) » hand grip strength test » Waist circumference » standing height » weight and bioimpedance » spirometry » pulse oximetry – Medical Outcomes Survey Short Form (SF-36v2) – Pain analogue scale – Fatigue scales (assessing severity and disability) – General Health Questionnaire (GHQ-28) – Epworth sleepiness score

20. Resources available Baseline laboratory test results, including: – full blood count – blood chemistry – liver function tests – erythrocyte sedimentation rate – c-reactive protein – rheumatoid factor – thyroid function tests – tissue transglutaminase antibodies – serum vitamin B12 – red cell folate

21. Who can access the samples? Researchers doing bona fide research Must submit application to UK ME/CFS Biobank Steering Group & UCL/RFH Biobank Samples can be selected from web-based portal

22. Procedures for request of samples Submission of application to ME/CFS Biobank project coordinator (Dr. Eliana Lacerda) Assessment by Steering Group Assessment by UCL RFH Biobank Committee Ethically approved research Anonymous aliquots provided to researchers

23. Currently planned research Immunology and genetics of ME/CFS LSHTM and UCL Virus presence and activity in ME/CFS LSHTM and Glasgow University

24. Thank you Contact us: Eliana Lacerda: eliana.lacerda@lshtm.ac.uk Erinna Bowman: erinna.bowman@lshtm.ac.uk Webpage: www.lshtm.ac.uk/mecfs