Suppressive Functions of Regulatory T Cells in Migraine

Overview

Migraine is a frequent, disabling condition, of great social and economical impact worldwide. This condition is more frequent in women and subjects with autoimmune and/or inflammatory diseases. Cytokine and immune cell dysregulations have been evidenced in migraine. Inflammation seems to play an important role in migraine chronification; however, the inflammatory mechanisms involved in migraine pathophysiology remain unclear. Regulatory T (Treg) cells play a central role in maintaining immune homeostasis. They regulate effector T (Teff) cell proliferation and cytokine production, through several suppressive mechanisms, such as the hydrolysis of adenosine triphosphate (ATP) into adenosine (ADO), mediated by surface enzymes Cluster Differentiation 39 (CD39) and Cluster Differentiation 73 (CD73). ATP is involved in pain processes in migraine, and insufficient hydrolysis could participate in pain chronification. Recent studies suggest altered proportions of Treg cells in migraine, and decreased levels of CD39-positive (CD39+) Treg cells, suggesting Treg suppressive functions may be decreased in the disease. However, there have been no functional studies to date to confirm this hypothesis. The investigators believe Treg suppressive functions may be decreased in migraine, and that such alterations may be caused by a malfunction in the ADO pathway.

Detailed description

Migraine is a major and global public health concern that affects 14% of the population worldwide. It is 2 to 3 times more prevalent in women than men and is the second leading cause of disability. Many patients remain unresponsive to currently available treatments. As evidenced in different studies, including our own, several auto-immune and inflammatory disorders, such as multiple sclerosis, are associated with a higher prevalence of migraine and a higher risk for migraine chronification (at least 15 headache days per month) compared to the general population. These findings suggest that inflammation may play an important role in migraine chronification. Cytokine and immune cell dysregulations have been evidenced in the disease but the role of inflammation in migraine pathophysiology remains unclear. Among T lymphocytes, regulatory T (Treg) cells play an important role in maintaining immune homeostasis. They regulate pro-inflammatory effector T (Teff) cells and proinflammatory cytokine release (IL-6, IL-17, interferon-γ) through different suppressive mechanisms such as the hydrolysis of pro-inflammatory and nociceptive adenosine triphosphate (ATP) into anti-inflammatory and antinociceptive adenosine (ADO), by Treg surface enzymes CD39 and CD73, present on 40% and 6-10% of Treg cells, respectively. ADO then suppresses Teff cells by binding to their ADO receptor A2A. ATP is involved in the transduction of pain signals in migraine and its insufficient hydrolysis can lead to pain chronification. It has been evidenced that Tregs naturally suppress interferon (IFN)-γ production by Teff cells. Their CD39-positive (CD39+) subsets also suppress IL-17 production by Teff cells, whereas CD39-negative (CD39-) Tregs seem to increase IL-17 secretion. Recent studies have shown altered Treg proportions in migraine compared to controls and Treg subpopulation dysregulations, such as decreased CD39+Treg cell levels. This further suggests that Treg suppressive functions may be altered in migraine, but no functional assays have been led to confirm that.

In this study, the investigators aim to determine whether Treg cells play a role in migraine pathophysiology, by answering the following 3 separate questions: Question 1) Are Treg suppressive functions decreased in migraine patients? Question 2) Are CD39+Treg cells functionally deficient in migraine patients? Question 3) Is the hydrolytic activity of enzyme CD39 reduced in migraine patients?

Through Treg/Teff coculture the investigators will assess the ability of total Treg and CD39+Treg cells to inhibit Teff proliferation and pro-inflammatory cytokine secretion and measure the hydrolytic activity of Treg enzyme CD39 on ATP in chronic migraine patients versus healthy controls. The use of human immune cells (Treg and Teff lymphocytes) in our experiments will allow the investigators to better understand migraine pathophysiology, with a direct application to human subjects, as well as to avoid animal testing and suffering. Through its animal-free setting, following the 'Reduce, Refine, Replace (3Rs)' principle, the investigators wish to promote animal-free research and the reduction of animal use for reagent production by using recombinant antibodies for flow cytometry and synthetic medium supplements for cell culture. This study will lead to a better understanding of the role of inflammation and Treg cells in migraine pathophysiology and will provide new perspectives for the development of personalized treatments according to the immune pain profile of migraine patients.

The investigators aim to show that Treg suppressive functions are altered in migraine and that the ADO pathway is deficient. Through Treg/Teff coculture the investigators will measure the ability of Tregs to inhibit Teff proliferation and cytokine secretion and assess the CD39-related hydrolytic activity of Tregs. This will allow the investigators to study Treg cells suppressive functions on Teff cells in female patients with chronic migraine and healthy female control participants.

Interventions

• Biological Blood sampling, questionnaire and phone call
  The blood sample will drawn after a fasting period of at least 8 hours, and the questionnaire will be auto-administered

Primary outcome measures

• Treg cell suppressive functions [Time frame: Day 0]

Secondary outcome measures (3)

• CD39+ Treg cell suppressive functions [Time frame: Day 0]
• CD39 activity [Time frame: Day 0]
• Cytokines [Time frame: Day 0]

Eligibility criteria

Inclusion criteria

• Females
• 18 to 50 years old
• Chronic migraine participants : chronic migraine (at least 15 days of headache/month, according to the International Classification of Headache Disorders, 3rd edition criteria

Exclusion criteria

• BMI < or = 17kg/m² or > or = 30kg/m²
• Diagnosis or suspicion of type 2 diabetes, auto-immune or inflammatory diseases, immunodeficiency diseases
• Diagnosis of headache of non-migraine origin, except for tension type headache < or = 4 days per month (i.e.: cluster headache, post-traumatic headache, cerebral tumour…)
• Pregnancy, delivery, miscarriage, breastfeeding, participation in a medically assisted human reproduction program (ovary stimulation/hormone therapy) < 3 months before blood sampling
• Menopause, hysterectomy, or bilateral oophorectomy
• Flare of the autoimmune/inflammatory disease of interest < 1 month before blood sampling
• Modification of maintenance therapy for the autoimmune/inflammatory disease of interest (start, change of molecule, interruption) < 3 months before blood sampling
• Modification of prophylactic anti-migraine therapy (start, change of molecule, interruption) < 3 months before blood sampling (for migraine participants)
• Hormone therapy (besides contraception and treatment of endometriosis)
• Transplantation (solid organ or bone marrow)
• Cancer (active or remitting) < 1 year before blood sampling (solid organ or blood)
• Haematologic disease (benign or malignant) of the lymphoid lineage
• Guardianship, curatorship, safeguard of justice or deprivation of liberty
• For controls : diagnosis of migraine

Criteria are shown verbatim from the registry (in English). Final eligibility is always assessed by the study center.

Healthy volunteers: Yes

Study design

Allocation

Non-randomized

Model

Parallel assignment

Masking

Open label

Primary purpose

Basic science

Study locations

France · 1 center

• CHU Clermont-Ferrand — Clermont-Ferrand

Publications

• Stovner LJ, Hagen K, Linde M, Steiner TJ. The global prevalence of headache: an update, with analysis of the influences of methodological factors on prevalence estimates. J Headache Pain. 2022 Apr 12;23(1):34. doi: 10.1186/s10194-022-01402-2. PMID 35410119
• Ashina M. Migraine. N Engl J Med. 2020 Nov 5;383(19):1866-1876. doi: 10.1056/NEJMra1915327. No abstract available. PMID 33211930
• Barbanti P, Aurilia C, Egeo G, Torelli P, Proietti S, Cevoli S, Bonassi S; Italian Migraine Registry study group. Late Response to Anti-CGRP Monoclonal Antibodies in Migraine: A Multicenter Prospective Observational Study. Neurology. 2023 Sep 12;101(11):482-488. doi: 10.1212/WNL.0000000000207292. Epub 2023 Apr 18. PMID 37072224
• Moisset X, Giraud P, Dallel R. Migraine in multiple sclerosis and other chronic inflammatory diseases. Rev Neurol (Paris). 2021 Sep;177(7):816-820. doi: 10.1016/j.neurol.2021.07.005. Epub 2021 Jul 27. PMID 34325914
• Moisset X, Ouchchane L, Guy N, Bayle DJ, Dallel R, Clavelou P. Migraine headaches and pain with neuropathic characteristics: comorbid conditions in patients with multiple sclerosis. Pain. 2013 Dec;154(12):2691-2699. doi: 10.1016/j.pain.2013.07.050. Epub 2013 Aug 2. PMID 23911697
• Edvinsson L, Haanes KA, Warfvinge K. Does inflammation have a role in migraine? Nat Rev Neurol. 2019 Aug;15(8):483-490. doi: 10.1038/s41582-019-0216-y. Epub 2019 Jul 1. PMID 31263254
• Thuraiaiyah J, Erritzoe-Jervild M, Al-Khazali HM, Schytz HW, Younis S. The role of cytokines in migraine: A systematic review. Cephalalgia. 2022 Dec;42(14):1565-1588. doi: 10.1177/03331024221118924. Epub 2022 Aug 12. PMID 35962530
• Nurkhametova D, Kudryavtsev I, Khayrutdinova O, Serebryakova M, Altunbaev R, Malm T, Giniatullin R. Purinergic Profiling of Regulatory T-cells in Patients With Episodic Migraine. Front Cell Neurosci. 2018 Sep 25;12:326. doi: 10.3389/fncel.2018.00326. eCollection 2018. PMID 30319363

Identifiers

NCT: NCT07067112 · RBHP 2025 MOISSET

Primary sources (government registries)

• ClinicalTrials.gov (US NIH)

View this study on ClinicalTrials.gov ↗