Biomarcadores de las formas clínicas de esclerosis múltiple: desarrollo de estrategias personalizadas basadas en perfiles inmunológicos de la vía del interferón de tipo I

Tesis inédita de la Universidad Complutense de Madrid, Facultad de Medicina, Departamento de Microbiología I, leída el 10/04/2013Multiple sclerosis (MS) is a highly heterogeneous disease and to date reliable plasma biomarkers that enable to distinguish among the different clinical forms of MS are lacking. The present work has identified blood biomarkers in 182 subjects (129 MS patients and 53 healthy controls) that were consecutively recruited as two independent cohorts. We found a significant lower expression of dipeptydil peptidase 4 (DPP4) and DPP activity in the plasma of MS patients with respect to healthy controls and DPP activity correlated inversely with clinical disability score in MS. Our results demonstrate that the divergent clinical and histology-based MS forms are associated with distinct profiles of circulating biomarkers, mostly chemokines and growth/angiogenic factors (HGF, Eotaxin/CCL11, MCP-1/CCL2, Rantes/CCL5, EGF, MIP-1β/CCL4, VEGF and FGFb); and with different gene expression levels in PBMCs of patients (CLU, IRF2 and LDLR). Responder patients to type I IFN displayed high levels of plasma IP10 and MCP-1, and a specific expression pattern of IFN stimulated genes.La esclerosis múltiple (EM) es una enfermedad muy heterogénea clínicamente que carece de biomarcadores plasmáticos que distingan entre formas clínicas. Este trabajo ha identificado biomarcadores de EM en sangre periférica en 182 sujetos (129 pacientes con EM y 53 controles sanos) reclutados en dos cohortes consecutivas. La expresión de dipeptidil peptidasa 4 (DPP4) y la actividad DPP están significativamente disminuidas en el plasma de nuestros pacientes con EM con respecto a los controles sanos y la actividad DPP se correlaciona inversamente con la escala de discapacidad clínica en EM. Nuestros resultados demuestran que las fases recurrente-remitente y la progresiva se asocian con distintos perfiles de biomarcadores circulantes, en su mayor parte quimiocinas y factores de crecimiento/angiogénicos (HGF, Eotaxin/CCL11, MCP-1/CCL2, Rantes/CCL5, EGF, MIP-1β/CCL4, VEGF y FGFb); y con diferentes niveles de expresión génica en sus PBMCs (CLU, IRF2 y LDLR). Los pacientes respondedores a IFN de tipo I presentan niveles elevados de IP10 y MCP-1 en plasma, así como una expresión específica de los genes estimulados por IFN.Depto. de Inmunología, Oftalmología y ORLFac. de MedicinaTRUEunpu

Multiple Sclerosis: Where do we go from here?

Multiple sclerosis (MS) is the most common cause of neurological disability in young populations after trauma and represents a significant personal, social, and economic public health burden. The clinical course and response of MS to therapy is highly heterogeneous, but most patients progress from a relapsing-remitting disease course, in which patients may respond to immunomodulatory drugs, to a steady progression and neurodegeneration that is unresponsive to any currently available treatment. In the last few years, novel disease-modifying therapies for MS have become available but the aetiology of the disease remains an enigma. The search for clinical biomarkers that are able to stratify MS patients and allow the personalisation of treatment strategies, has developed greatly in recent years though only a few have been integrated into routine clinical practice

Perforin Expression by CD4+ Regulatory T Cells Increases at Multiple Sclerosis Relapse: Sex Differences

Abstract: Multiple sclerosis (MS) represents the leading cause of neurological deficit among young adults, affecting women more frequently than men. In MS, the extent of central nervous system lesions is determined by the net balance between self-reactive and regulatory T-cells at any given time, among other factors, as well as by the effect of inflammatory response. Here, we studied both CD4+ and CD8+ TReg in parallel in blood and CSF during MS relapse. A recruitment of both regulatory CD4+ and CD8+ T cells (TReg) within the cerebrospinal fluid (CSF) takes place during MS relapse. Not previously described, the presence of CD4+ TReg in CSF was higher in women than in men, which could account for the sexual dimorphism in the incidence of MS. A direct correlation between plasma oestradiol (E2) and IL-2 levels was observed, in line with a putative circuit of E2 and perforin expression by CD4+ TReg playing a role in MS. Also, serum IFN-alpha was higher in females, with direct correlation with serum E2 levels. This is the first study to analyze perforin expression by CD4+ TReg in MS, which was greatly enhanced in CSF

Low DPP4 expression and activity in multiple sclerosis

International audienceMultiple sclerosis (MS) is a prototypic Th1/Th17 chronic autoimmune disease of the central nervous system. Dipeptidyl peptidase 4 (DPP4 or CD26) is a multifunctional molecule involved in autoimmune diseases' pathophysiology. We sought to integrate disparate pieces of data and analyze the plasma levels of sDPP4, DPP activity and DPP4 surface expression on T-cells in 129 MS patients with different clinical forms and 53 healthy controls, across two independent cohorts. Herein, we provide new evidence that sDPP4 concentration and DPP activity are significantly lower in MS patients than controls (p < 0.0001 and p < 0.01, respectively). In contrast, the frequency of circulating CD8(+)DPP4(hi) T-cells (p = 0.02) was increased in MS patients. This is the first study that simultaneously analyzes DPP4 expression and function in a large cohort of MS patients. Our data indicate a putative role for DPP4 in MS pathophysiology and suggest that a deeper understanding of surface versus shed DPP4 biology is warranted

HGF, Eotaxin/CCL11, MCP-1/CCL2, Rantes/CCL5, EGF and MIP-1β/CCL4 comparison in the test and validation cohorts.

<p>Mann—Whitney statistical test was used for calculation of the reported p-value between RR-MS vs Progressive MS; RR-MS = Relapsing-Remitting MS patients. Note that the Relapsing-remitting group comprises the clinical groups: RR-MS Remission, RR-MS Active, RESPONDERS, NON RESPONDERS and RELAPSES. The Progressive group comprises Secondary (SP-MS) and Primary-progressive patients.</p><p>Results are given as Median value (IQR1–3). Statistical significance is marked as:</p><p>*p<0.05;</p><p>**p<0.01;</p><p>***p<0.001.</p><p>^# p = 0.06 (trend).</p><p>HGF, Eotaxin/CCL11, MCP-1/CCL2, Rantes/CCL5, EGF and MIP-1β/CCL4 comparison in the test and validation cohorts.</p

IP10/CXCL10 and MCP-1/CCL2 levels were higher in MS patients with good response to IFN-β treatment.

<p>Long-term IFN-β treated RR-MS patients (RESPONDERS, n = 20) presented with higher circulating levels of IP10/CXCL10 and MCP-1/CCL2 than non responders to IFN-β (NON RESPONDERS, n = 13). Mann—Whitney statistical test was used for calculation of the reported p-value; median values are represented by a gray bar; individual dots indicate single donor values.</p

VEGF levels were significantly higher in secondary progressive MS patients.

<p>Plasma levels of the vascular growth factor (VEGF) in the validation cohort, were higher in secondary progressive MS patients (SP-MS, n = 16) than in patients with the relapsing-remitting form (RR-MS, n = 38) and than in healthy controls (HC, n = 20). The SP-MS patients had higher plasma levels of VEGF than primary progressive MS patients (PP-MS, n = 9) but this difference did not reach statistical significance. Mann—Whitney statistical test was used for calculation of the reported p-value; median values are represented by a gray bar; individual dots indicate single donor values.</p

Biomarkers that discriminate between Relapsing-Remitting and Progressive Clinical forms of MS by ROC curves.

<p>The areas under the curve (AUC), 95% Confidence Interval (95% CI) and p value for each analyte are depicted in Table 2.</p><p>Biomarkers that discriminate between Relapsing-Remitting and Progressive Clinical forms of MS by ROC curves.</p

Different plasma levels of EGF and MIP-1β/CCL4 in MS clinical forms.

<p>Plasma levels of EGF and MIP-1β/CCL4 were lower in the progressive MS than in relapsing-remitting MS patients. Comparison of the plasma levels of EGF and MIP-1β/CCL4 between patients with relapsing-remitting (RR-MS, n = 80) and in progressive MS patients (n = 49). Mann—Whitney statistical test was used for calculation of the reported p-value; median values are represented by a gray bar; individual dots indicate single donor values. Note that the Relapsing-remitting group comprises the clinical groups: RR-MS Remission, RR-MS Active, RESPONDERS, NON RESPONDERS and RELAPSES. The Progressive group comprises Secondary (SP-MS) and Primary-progressive patients.</p