Title: Chief Scientist Office / Action for M.E. funded Ph.D. in biomedical research into Myalgic Encephalomyelitis.
Funder: Chief Scientist Office, Scotland

Abstract: Myalgic Encephalomyelitis (ME; otherwise known as Chronic Fatigue Syndrome [CFS]), is a long term illness of unknown origin with debilitating symptoms [1]. Those with ME/CFS experience a wide variety of physical and cognitive symptoms, but the illness is mainly characterised by post-exertional malaise - extreme exhaustion after any kind of physical activity that is not improved by resting [1]. Studies have implied that the quality of life for the average patient is worse than that for most other illnesses, including some cancers [2], and have found that it affects a large number of people (estimated at a minimum 0.2% of the population) [3]. There is no test or clear marker for ME/CFS, and so clinicians are required to exclude other possibilities with similar symptoms before reaching a diagnosis [1]. The search for a reliable biomarker is a major component of current research efforts. Recently, Dr Mark Davis (Stanford University) presented unpublished results comparing the T-cells of ME/CFS patients versus both healthy and ill controls. He found that the former had significantly increased clonal expansion compared to the healthy controls. This type of immune activation is typically seen in patients with other conditions, and the ME/CFS results were in line with those seen in the ill controls (with multiple sclerosis and Lyme disease). For our experiments and analysis, we will be aligning our methods with those of Systems Biology Laboratory (SBL), Oxfordshire. In this collaboration we will use a method of library preparation and sequencing (T-cell diversity assay), to count the number of unique T-cell receptor (TCR) types (αδ, β and γ; T-cell receptor “clonotypes”) from 40 females in each of four groups. These are: 1) Severely affected ME/CFS cases – those who are either bed- or house-bound; 2) Mild/moderate ME/CFS cases; 3) Disease controls – multiple sclerosis (MS); and 4) Healthy (physically active) controls. We will estimate the degree of TCR clonality (defined as 1 - Pielou’s Evenness) for each person in the experiment [4][5]. As we are measuring for each of the three TCRAD, TCRB and TCRG loci, we will obtain three values per sample which are expected to show high correlation. If so, then these three values will be averaged to provide a single clonality measure. We will then compare the statistics of this measure for each subgroup, via six pairwise Mann-Whitney U tests. We shall also undertake a supplemental analysis, using instead the Shannon entropy (from which we will have derived Pielou’s Evenness) and Simpson’s diversity index. Null Hypothesis H0: Our null hypothesis is that there is no significant difference in the distributions of clonality between any pair of the four subgroups. Alternate Hypothesis H1: We hypothesise that the distribution of the clonality of the cases and MS controls will be greater than that of the healthy controls. Once we have tested this hypothesis, we will test for correlations between each individual’s clonality value and:  The physical component of the SF-36v2 questionnaire (which has been validated as a good metric for ME severity) [6][7];  A pain/fatigue scale; and  Handgrip strength, which has been shown (although not yet validated) to correlate with ME severity [8].


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: DISPUTED AETIOLOGY AND OTHER