Role of the Nuclear Pore Component RANBP2 in Inflammatory Responses to Viral Infections

Not Applicable

Not yet recruiting

• Conditions: Rare Genetic Disease; Acute Necrotizing Encephalopathy

• Registration Number: NCT06731790
• Lead Sponsor: University Hospital, Montpellier

Brief Summary

The goal of this controlled, pathophysiological, exploratory interventional study is to compare the inflammatory phenotype of circulating immune cells, basal and following stimulation, from Acute Necrotizing Encephalopathy Type 1 (ANE1) patients with those from sex- and age-matched donors who do not carry the mutation.To date, no study has investigated the molecular mechanisms regulating the inflammatory response in ANE1 disease directly on patient samples.

The primary endpoint in individuals in the "mutated RANBP2" arm is an inflammatory phenotype (hyperinflammatory monocytes, secretion of pro-inflammatory cytokines, anti-glycoprotein autoantibodies), significantly exacerbated basal and/or post-stimulation production of pro-inflammatory cytokines compared with the control arm.

The secondary objective is to examine the allelic expression of mutant RANBP2 and characterize genetic variants by whole-exome sequencing, in order to associate them with RANBP2 protein localization and ANE crisis severity

The researchers will compare the group of ANE1 patients with age- and sex-matched control groups, divided into two subgroups: unrelated controls and controls with familial ties. The aim is to study the different types of inflammatory responses and correlate them with the localization of the RANBP2 protein and the severity of ANE episodes.

Participants will participate in a single visit during which demographic data, clinical history and a blood test will be collected with one (unrelated control) or two blood tubes (ANE1 and related control).

Detailed Description

The nucleoporin RANBP2, also known as Nup358, is a component of the cytoplasmic filaments of nuclear pore complexes (NPCs), which regulate the transport of macromolecules between the cytoplasm and the nucleus. Mutations in the RANBP2 gene are associated with a rare genetic predisposition to acute necrotizing encephalopathy (ANE1), a predominantly pediatric disease characterized by multiple, symmetrical hemorrhagic lesions of the brain following febrile infection, most often with influenza A virus (IAV). Given the presumed inflammatory nature of ANE1, first-line treatment includes intravenous administration of high-dose pharmacological corticosteroids with or without immunoglobulins. In addition, two clinical cases report that IL-6 inhibition by tocilumizab may have a beneficial role.

the research team have recently demonstrated that IAV infection disrupts the localization of RANBP2 and key inflammatory mediators such as inflammasome components. We have also shown that RANBP2 regulates the expression of pro-inflammatory cytokines at both transcriptional and post-translational levels.

The researchers hypothesis is that mutation of RANBP2 in ANE1 patients weakens nuclear pore control of innate immune signaling pathways, leading to an exacerbated inflammatory response to infections.

Study Timeline

• Status Verified: 2024-11
• Study First Submitted: 2024-12-09
• Study First Submitted QC: 2024-12-09
• Last Update Submitted: 2024-12-09
• Study First Posted: 2024-12-12
• Last Update Posted: 2024-12-12
• Study Start: 2025-01-01
• Primary Completion: 2026-07-01
• Study Completion: 2026-07-01

Recruitment & Eligibility

• Status: NOT_YET_RECRUITING
• Sex: All
• Target Recruitment: 64

Inclusion Criteria

• Obtaining written consent from adult participants
• Obtaining written consent from legal guardians of minors with their assent
• Subjects aged 1 to 90
• Subject with a T585M mutation in RANBP2 for the RANBP2 mutation arm.
• Subjects matched on age (+/- 10 years) and gender for the "control" arm.

Exclusion Criteria

• Patient not affiliated to a social security scheme or not a beneficiary of such a scheme. (for example, a European Health Insurance Card (EHIC))
• Absence of written informed consent
• Person unable to give consent
• legally protected adult (guardianship, curatorship)
• Person deprived of liberty
• Person participating in another research study with an exclusion period still in progress
• Chronic or infectious disease affecting the immune system
• Immunomodulating treatment (e.g. corticosteroids, immunoglobulins)

Study & Design

• Study Type: INTERVENTIONAL
• Study Design: PARALLEL

Primary Outcome Measures

Name	Time	Method
Comparison of the inflammatory phenotype of circulating immune cells, both basal and following stimulation, from ANE1 patients with those from sex- and age-matched donors not carrying the mutation	Baseline	These experiments will be carried out on blood, serum, peripheral blood mononuclear cells (PBMC) and monocyte-derived macrophages and microglia (MDM and MMG, respectively).; The primary endpoint for individuals in the "mutated RANBP2" arm is an inflammatory phenotype, basal and/or after stimulation, that is significantly exacerbated compared with the "control" arms. Samples will be analyzed by ELISA/Luminex and flow cytometry. The expected difference will be a 2 to 10-fold upregulation in one or several markers of hyperinflammatory monocytes, and/or in one or several secreted pro-inflammatory cytokines, and/or in one or several autoantibodies, based on our results in PBMC treated with RANBP2-targeted RNA interference

Secondary Outcome Measures

Name	Time	Method
Determination of the allelic expression of mutated RANBP2	Baseline	The first secondary endpoint will be heterozygous expression of mutated RANBP2 in individuals in the "mutated RANBP2" arm. The method of evaluation will be RT-PCR targeted to RANBP2, followed by quantitative PCR discriminating for the mutation, a diagnostic method recently introduced in the IRIM laboratory.
Determination of the effect of the T585M heterozygous mutation on RANBP2 localization	Baseline	The second secondary outcome will be the significantly exacerbated basal and/or post-stimulation RANBP2 delocalization compared with the "control" arm. This will be tested on monocyte-derived macrophages by confocal microscopy.
Characterization of genetic variants that are significantly associated with ANE seizure severity	Baseline	The third secondary endpoint will be the identification of genetic variants associated with the history of ANE episodes, if any, and their severity. The evaluation method will be whole exome sequencing, only on participants from ANE families in the "control" and "mutated RANBP2" arms.

Trial Locations

Locations (1)

CHU Gui de Chauliac; 🇫🇷 Montpellier, Hérault, France