5-HT7 Receptor Implication in Inflammatory Mechanisms in Multiple Sclerosis

NCT04546698 | Completed

• 1 other identifier: CHRO-2020-01
• Study Type: observational
• Target: 78
• Locations: 1 country
• Sites: 1

Brief Summary

Multiple Sclerosis is a chronic autoimmune disease associated with inflammatory response harmful for the Central Nervous System. Immunological imbalance is involved with Th1 and Th17 cells in correlation with a disturbance of regulators mechanisms as Treg cells. Despite years of research, the mechanisms involved remain unclear. Serotonin (5-HT) seems to be a therapeutic target to treat multiple sclerosis. Indeed, several studies have shown the anti-inflammatory potential of this neurotransmitter and also its vulnerability in inflammatory context. Moreover, a recent study has shown that 5-HT can reduced CD4 T cells proliferation and pro-inflammatory cytokines released in vitro. 5-HT protector effects have also demonstrated in Experimental Autoimmune Encephalomyelitis mouse model (EAE) with an inflammatory response reduction and also a decreased of spinal cord lesions. The latest receptor discovered, the 5-HT7 receptor, has been identify as a promise target to treat neurological disorders associated with inflammatory context. Present in humans and mice, this receptor spreads on the surface of a large number of cells, such as T-lymphocytes, macrophages, dendritic cells and also neurons, astrocytes and microglia. Given the importance of the positive cells for 5-HT7 receptor, in the inflammatory context observed in multiple sclerosis, The investigator propose to study the receptor expression in blood samples from multiple sclerosis patient.

Conditions

Multiple Sclerosis, Acute Relapsing; Multiple Sclerosis

Keywords

Multiple sclerosis; serotonin; 5-HT7 receptor; inflammation; lymphocytes; cytokines

Outcome Measures

Primary Outcomes (5)

• 5HT7 Serotonin receptor expression on circulating leukocyte cells

  To analyze the 5-HT7 expression on circulating leukocyte cells, we will perform flow cytometry studies from the whole blood of the three groups mentioned above. In a first step, we will analyze the overall rate of 5-HT7 circulating positive cells by the use of an antibody against 5-HT7 receptor whose specificity was validated in our research team. In a second step, we will precise which cell type, among the lymphocytes implicated in the MS pathology, express the 5-HT7 receptor. We will use antibodies against each population: Thelper 1 cell, Thelper 2 cell, Thelper 17 cell and Treg. We will also perform immunostaining on Cytospin (thin layer of blood cells on coverslips) to visualize co-labeling and confirm the results of flow cytometry.

  Baseline

• 5-HT7 Serotonin receptor mRNA quantity

  For the three groups, we will determine the 5-HT7 mRNA quantity on circulating leukocyte cells. In a first step, we will check correlation between the 5-HT7 receptor expression rate and the mRNA quantity on circulating leukocyte cells, using quantitative Polymerase Chain Reaction (qPCR). In a second step, we will perform cell sorting to isolate Thelper 1 cell, Thelper 2 cell , Thelper 17 cell and Treg cells and specifically extract 5-HT7 mRNA.

  Baseline

• Immunological context on blood samples

  We will identify the immune context by ELISA dosage from serum samples. We will investigate on pro-inflammatory markers like Interleukin 1b, IFNg, IL17 and anti-inflammatory markers like Interleukin 10 and Interluekin 4.

  Baseline

• 5-HT7 receptor role on lymphocyte functions in vitro

  From blood samples of the three groups mentioned above, we will isolate peripheral blood mononuclear cells (PBMC) in vitro, to evaluate the 5-HT7 receptor role on their function to produce and release cytokines. We will stimulate PBMC with one or more 5-HT7 ligands, and we will perform ELISA dosage from supernatant with pro-inflammatory markers like IL1b, IFNg, IL17 and anti-inflammatory markers like IL10 and IL4. The cell immunolabelling will be perform to visualize 5-HT7 receptor on the cell surface.

  Baseline

• 5-HT7 receptor isoforms 5-HT7 receptor isoforms

  There are three forms of 5-HT7 receptor (5-HT7a, 5-HT7b, 5-HT7d). We propose to identify these different isoforms by qPCR. As previously described, in a first step, on circulating leukocyte cells and in a second step, after cell sorting to isolate Th1, Th2, Th17 and Treg cells.

  Baseline

Study Arms (3)

Multiple Sclerosis patients with an acute relapse

Multiple Sclerosis diagnosed according to Mc Donald's criteria with an acute relapse

Other: blood sampling

Multiple sclerosis pataients treated with Natalizumab

Multiple Sclerosis patients diagnosed according to Mc Donald's criteria and treated with Natalizumab since 6 cures

Other: blood sampling

Healthy people

Other: blood sampling

Interventions

blood sampling OTHER

Analyse of expression modulation of 5-HT7 receptor in correlation with the immunological context, in 3 differents groups : 2 with multiple sclerosis patients, one healthy volunteers group

Healthy people; Multiple Sclerosis patients with an acute relapse; Multiple sclerosis pataients treated with Natalizumab

Eligibility Criteria

• Age: 18 Years - 50 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)
• Sampling Method: Non-Probability Sample

Study Population

Man and Woman 18 to 50 years old

You may qualify if:

• Man and Woman
• to 50 years old
• Multiple Sclerosis diagnosed according to Mc Donald's criteria (Thompson et al. 2018)
• With an acute relapse (group 1)
• Stable with Natalizumab treatment (group 2), minimum 6 Natalizumab cures
• Healthy Volunteers (group 3)

You may not qualify if:

• without social security
• HIV positive serology
• infectious status in the past month
• corticosteroid therapy in the past month
• dementia
• pregnant or breastfeeding woman
• previous participation in the study
• under judicial protection
• non-cooperating patient

Sponsors & Collaborators

• Centre Hospitalier Régional d'Orléans: lead
• National Scientific Research Centre: collaborator

Study Sites (1)

CHR Orléans

Orléans, 45067, France

Location

Related Publications (4)

• Thompson AJ, Banwell BL, Barkhof F, Carroll WM, Coetzee T, Comi G, Correale J, Fazekas F, Filippi M, Freedman MS, Fujihara K, Galetta SL, Hartung HP, Kappos L, Lublin FD, Marrie RA, Miller AE, Miller DH, Montalban X, Mowry EM, Sorensen PS, Tintore M, Traboulsee AL, Trojano M, Uitdehaag BMJ, Vukusic S, Waubant E, Weinshenker BG, Reingold SC, Cohen JA. Diagnosis of multiple sclerosis: 2017 revisions of the McDonald criteria. Lancet Neurol. 2018 Feb;17(2):162-173. doi: 10.1016/S1474-4422(17)30470-2. Epub 2017 Dec 21.

  PMID: 29275977 BACKGROUND

• Sacramento PM, Monteiro C, Dias ASO, Kasahara TM, Ferreira TB, Hygino J, Wing AC, Andrade RM, Rueda F, Sales MC, Vasconcelos CC, Bento CAM. Serotonin decreases the production of Th1/Th17 cytokines and elevates the frequency of regulatory CD4+ T-cell subsets in multiple sclerosis patients. Eur J Immunol. 2018 Aug;48(8):1376-1388. doi: 10.1002/eji.201847525. Epub 2018 Jun 6.

  PMID: 29719048 RESULT

• Yuan XQ, Qiu G, Liu XJ, Liu S, Wu Y, Wang X, Lu T. Fluoxetine promotes remission in acute experimental autoimmune encephalomyelitis in rats. Neuroimmunomodulation. 2012;19(4):201-8. doi: 10.1159/000334095. Epub 2012 Mar 21.

  PMID: 22441536 RESULT

• Quintero-Villegas A, Valdes-Ferrer SI. Role of 5-HT7 receptors in the immune system in health and disease. Mol Med. 2019 Dec 31;26(1):2. doi: 10.1186/s10020-019-0126-x.

  PMID: 31892309 RESULT

Biospecimen

Retention: SAMPLES WITHOUT DNA

one sampling to assess nvestigate the expression modulation of 5-HT7 receptor

MeSH Terms

Conditions

Multiple Sclerosis, Relapsing-Remitting; Multiple Sclerosis; Inflammation

Interventions

Blood Specimen Collection

Condition Hierarchy (Ancestors)

Demyelinating Autoimmune Diseases, CNS; Autoimmune Diseases of the Nervous System; Nervous System Diseases; Demyelinating Diseases; Autoimmune Diseases; Immune System Diseases; Pathologic Processes; Pathological Conditions, Signs and Symptoms

Intervention Hierarchy (Ancestors)

Specimen Handling; Clinical Laboratory Techniques; Diagnostic Techniques and Procedures; Diagnosis; Punctures; Surgical Procedures, Operative; Investigative Techniques

Study Officials

• Maud PALLIX, MD

  CHR ORLEANS

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: observational
• Observational Model: CASE CONTROL
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: August 31, 2020
• First Posted: September 14, 2020
• Study Start: September 7, 2020
• Primary Completion: July 19, 2021
• Study Completion: July 19, 2021
• Last Updated: February 17, 2022

Record last verified: 2022-02

Data Sharing

• IPD Sharing: Will not share

Locations

FR (1)