Instituto de Investigación Sanitaria del Hospital Clínico San Carlos (IdISSC)
Environmental factors in degenerative diseases research group

Author Of 6 Presentations

Biomarkers and Bioinformatics Poster Presentation

P0109 - microRNA expression and its association with disability and brain atrophy (ID 1255)

Abstract

Background

MicroRNAs (miRNAs) are endogenous, non coding, small RNAs with post-transcriptional regulating functions. They participate in several cellular processes, including inflammation, neurodegeneration and remyelination

Objectives

To correlate serum miRNAs profile expression with disability, cognitive functioning and brain volume in patients with remitting-relapsing multiple sclerosis (RRMS)

Methods

Cross-sectional study in RRMS patients on stable treatment with glatiramer acetate (GA) during at least 6 months.

We selected the 20 best miRNAs candidates for RRMS and cognitive dysfunction through simple topological analysis (Anaxomics ®). MiRNAs profile was determined with LNA-based PCR (Exiqon). Clinical variables were Expanded Disability Status Scale (EDSS), Symbol Digit Modalities Test (SDMT) and brain volume (whole brain volume, grey matter volume, white matter volume, cerebellum volume, basal ganglia volume and T1 lesion load volume) (automatic software NeuroQuant ®). Correlation was analyzed with Spearman correlation coefficient (r) (p<0,05; software: SPSS)

Results

We included 20 patients (13 women), age 38,2 (29,4, 47,8) years, duration of disease: 5,1 (1,5, 8,5) years, and time on GA 2,1 (0,8, 6) years. We found a pathogenic association between miR.146a.5p and has.mir.9.5p with EDSS (r:0,434, p=0,03; r:0,516, p=0,028); miR.146a.5p with SDMT (r:-0,476, p=0,016); has.mir.9.5p with thalamus (r:-0,545, p=0,036), and miR.200c.3p with pallidum and cerebellum (r:-0,675, p=0,002; r:-0,472, p=0,048).

Conclusions

MiRNAs could be useful biomarkers in multiple sclerosis. We would like to highlight the association of proinflammatory has.mir.9.5p with EDSS and thalamus volume. They are needed more studies to confirm this findings.

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Metabolomics Poster Presentation

P0514 - Analysis of short chain fatty acids levels in serum and CSF samples of patients with multiple sclerosis and other neurological diseases (ID 1204)

Presentation Number
P0514
Presentation Topic
Metabolomics

Abstract

Background

In the recent years, gut microbiota has been related to multiple sclerosis (MS) etiopathogenesis. Short-chain fatty acids (SCFA) are metabolites derived from microbial metabolism that could play a fundamental role in gut-brain axis. The most abundant SCFA are butyrate, propionate and acetate (more than 95%). There is a controversy with the role of these SCFA in relation to the immune system. While butyrate and propionate have anti-inflammatory properties, it has been proposed that acetate could be a pro-inflammatory metabolite. In a previous study of our group, acetate was found elevated in serum samples of MS patients in comparison with healthy controls.

Objectives

To analyze the levels of acetate, propionate and butyrate in cerebrospinal fluid (CSF) and serum samples of patients with MS and other neurological diseases (OND).

Methods

Sixty-seven serum samples were collected (39 from MS patients, 30 at the time of clinically isolated syndrome [CIS] and 9 from patients with relapsing-remitting MS [RRMS], and 28 from patients with OND). For 41 of these patients we also had a CSF sample collected at the same time (31 from MS patients, 23 at the time of CIS and 8 from patients with RRMS, and 28 from patients with OND). We analyzed in these samples the levels of acetate, propionate and butyrate by liquid chromatography-mass spectrometry.

Results

Serum acetate was significantly elevated in MS patients compared to OND patients (30.3 mM vs. 11.7 mM, respectively; p=0.002). Butyrate was significantly lower in the serum of MS patients (p = 0.026). Acetate / propionate and acetate / butyrate ratios were significantly elevated in serum samples of MS patients (p <0.0001, in both cases). We did not find significant differences between MS and OND when we analyzed the levels of acetate, butyrate and propionate in CSF samples.

Conclusions

In this study, acetate was found elevated in serum samples of MS patients in comparison to patients with OND. The ratios between acetate and propionate or butyrate (both anti-inflammatory metabolites), could help us to discriminate between MS patients and patients with OND. Larger cohorts are needed to study the possible relevance of SCFA in CSF samples.

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Pathogenesis – Immunology Poster Presentation

P0939 - Antibody titters against EBV and HHV-6A/B and expression of MSRV ENV in the serum of pregnant multiple sclerosis patients (ID 1197)

Abstract

Background

Pregnancy is a special period within the clinical course of multiple sclerosis (MS), characterized by a reduction in the relapse rate and slower disease progression. On the contrary, during puerperium, relapse rate increases again. Viruses have been related to the etiopathogenesis of the disease, especially with disease activity.

Objectives

To analyse the serum antibody titters against Epstein-Barr virus (EBV) (EBNA-1 and VCA) and human herpesvirus 6 A/B (HHV-6A/B), as well as the expression of the envelope protein of the MS-associated retrovirus (MSRV ENV) in pregnant MS patients during pregnancy and postpartum. To study their possible relationship with the disease activity during pregnancy and postpartum, as well as their potential role in predicting the risk of relapses.

Methods

Serum samples were collected from 71 pregnant women, 50 with MS and 21 healthy controls, at every trimester of pregnancy and in the postpartum. Antibody titters against the above mentioned viruses were analysed by ELISA commercial kits, following manufacturer instructions; gene expression of MSRV ENV was analysed by qRT-PCR.

Results

IgM titres against HHV-6A/B were higher in MS patients than in healthy controls in the three trimesters of pregnancy and in the postpartum period (U-Mann Whitney): p =0.00001 for the first trimester; p=0.021 for the second trimester; p = 0.000005 for the third trimester; p =0.001, for the postpartum period). Furthermore, IgM titres against HHV-6A/B in the first trimester were higher in patients with relapses (U Mann Whitney, p = 0.052). Regarding the expression of MSRV ENV, the percentage of positivity during the first trimester was significantly higher in MS patients with relapses during pregnancy compared to those who did not (Fisher, p = 0.038).

Conclusions

High IgM titters against HHV-6A/B and the expression of MSRV ENV during the first trimester of pregnancy could act as predictors of relapse risk during pregnancy / postpartum. Although further studies are needed to validate these results, this study support the relation between viruses and relapses in pregnant MS patients.

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Pathogenesis – Immunology Poster Presentation

P0964 - Herpesvirus serology in primary progressive multiple sclerosis (ID 919)

Abstract

Background

Viruses have been involved in multiple sclerosis (MS) in last years. However, almost all of the studies published so far have focused on patients with relapsing-remitting MS (RRMS), with no data about viruses in patients with progressive primary MS (PPMS). Due to the differences that exist between these two forms of MS it would be interesting to know if there is also differences regarding viruses previously related to the etiopathogenesis of the disease.

Objectives

The aim of this study was to analyze the prevalence and titters of different viral antibodies: IgG against EBNA-1 and VCA of Epstein-Barr virus (EBV), IgG and IgM against Human herpesvirus 6 (HHV-6) and cytomegalovirus (CMV), in PPMS patients and to compare those results with other cohort of RRMS patients.

Methods

A total of 166 MS patients were recruited: 71 with PPMS and 95 with RRMS (mean age: 48 and 43.6 years, respectively; gender: 47.9% and 61% female, respectively). We analyzed the presence and titters of the antibodies above mentioned with ELISA commercial kits, following manufacturer instructions.

Results

1. Herpesvirus prevalences: we only found a statistical significant difference for IgG antibodies against EBNA-1 of EBV (84.5% of PPMS patients were positive vs. 97.8% of RRMS patients; p=0.002). When we analyzed these prevalences by age groups (<45 and >45 years), we only found statistical significant differences between PPMS and RRMS patients under 45 years: 84.6% of PPMS patients were positive for IgG against EBNA-1 vs. 100% of RRMS patients (p=0.015), and 26.9% of PPMS patients were positive for IgM against HHV-6 vs. 7.7% of RRMS patients (p=0.021). 2. Herpesvirus titters: we found that IgG titters against CMV were higher among PPMS patients than in RRMS patients (31.3 AU vs. 16.01 AU, p=0.0001); IgG titters against HHV-6 were also higher in PPMS patients (29.3 AU vs. 20.5, p=0.004); IgG titters against EBNA-1 of EBV were lower in PPMS patients than in RRMS patients (16.4 AU vs. 22.8 AU, p<0.0001).

Conclusions

We have found statistical significant differences between different herpesvirus prevalences and titters between PPMS and RRMS patients that should be deeper studied to evaluate their possible contribution to the existing differences between these two forms of MS.

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Pathogenesis – Immunology Poster Presentation

P0975 - Lipid-specific oligoclonal IgM bands condition age-related changes in multiple sclerosis (ID 1396)

Abstract

Background

Age has high influence on clinical course of chronic inflammatory diseases such as multiple sclerosis (MS). Lipid-specific oligoclonal IgM bands (LS-OCBM) in cerebrospinal fluid (CSF) of MS patients are biomarkers of high inflammatory disease. However, the relationship between both variables has not been fully studied.

Objectives

To explore age-related changes in CSF of MS patients leading to senescence and to study the role of LS-OCBM in this process.

Methods

A prospective cross-sectional study of 263 MS patients followed at Ramon y Cajal University Hospital, Madrid. Seventy-two showed (M+) and 191 (M-) lacked CSF LS-OCBM. We explored CSF immune cells, soluble mediators, microRNAs and clinical data.

Results

MS patients older than 45 years (O45) showed significant rises in CSF Programmed death ligand 1 (PD-L1; p=0.001) and T-cell immunoglobulin mucin-3 (TIM-3; p<0.0001), which induce lymphocyte tolerance, and in Chitinase 3-like-1 (p<0.0001) and Activin-A (p<0.0001), which lead to innate cell activation. This associated with low values of pro-inflammatory B cells and of Th17 lymphocytes. In addition, M- O45 patients experienced significant increases in miR-125b-5p (p=0.01) and miR-145-5p (p=0.02), two miRNA associated with immune senescence, an increase of let-7b-5p (p=0.01) that counteract innate-cell activation, a dramatic decrease of CSF lymphocytes (p<0.0001) and of NK cells (p=0.002) and increased serum anti-cytomegalovirus antibodies (p=0.009). However, M+ O45 patients exhibited a reduction of miR-204-5p (p=0.01), that induce innate-cell activation and increased CSF levels of superoxide dismutase (p=0.04) and isoprostane (p=0.03), markers of oxidative stress. These changes resulted in increased Multiple Sclerosis Severity Score (MSSS) on M+ O45 patients.

Conclusions

M- O45 MS patients showed inhibition of the intrathecal adaptive immune response and early immunosenescence. Conversely, M+ O45 MS patients displayed intrathecal innate cell activation and accelerated disability worsening. These data could be relevant for treating aged MS patients.

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Reproductive Aspects and Pregnancy Poster Presentation

P1138 - short-chain fatty acid during pregnancy in multiple sclerosis: a prospective cohort study (ID 1801)

Abstract

Background

Objective: Pregnancy reduces the annualized relapse rate (ARR) in multiple sclerosis (MS). However it is temporarily increased at puerperium. The exact mechanism underlying this clinical observation remains unknown.

Objectives

We aimed to explore the changes of the short-chain fatty acids (SCFAs) profiles in MS patients and healthy women (HCW) during pregnancy, and to assess any potential biomarker predicting the appearance of relapses during pregnancy and postpartum.

Methods

Methods: We included 53 pregnant MS patients and 21 HCW followed at Hospital Universitario Gregorio Marañón between 2007 and 2018. Patients were evaluated at every trimester of pregnancy and in the puerperium. Serum SCFAs were measured by liquid chromatography-mass spectrometry

Results

Results: Seventeen patients (32%) experienced relapses during pregnancy or puerperium (ACT group) and 37(68%) did not (NO-ACT group). We did not found differences in clinical characteristics or treatment status between the two groups. We observed differences in the SCFAs profile between ACT:NO-ACT groups during pregnancy and puerperium. Acetate levels were higher during the pregnancy-puerperium period in MS patients, regardless of clinical activity, compared to HCW (p: 0.0001). Interestingly, levels of propionate and butyrate below 1,985 μM (1,283 - 3.55) and 0.515 μM (0.01-2.943) respectively during the first trimester were associated with relapses during the pregnancy-puerperium period (p: 0.0001). The ACT group had a lower ratio of propionate/acetate and butyrate/acetate during pregnancy compared to NO-ACT group (<0.0001). The ROC curve showed that a propionate/acetate ratio of 0.36 (AUC 0.96, sensitivity 94%, specificity 92%;) has an OR=165 [CI: 10.2-239.4], p <0.0001) to predict relapses during pregnancy-puerperium.

Conclusions

Conclusions: SCFA levels during pregnancy may be associated with clinical activity in MS, and that their measurement could be useful in predicting the occurrence of relapses during this pregnancy-puerperium period

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