Ignite Creation Date:
2024-05-06 @ 5:15 PM
Last Modification Date:
2024-10-26 @ 2:25 PM
Study NCT ID:
NCT05245344
Status:
UNKNOWN
Last Update Posted:
2022-02-17
First Post:
2021-12-30
Brief Title:
Effects of Ozanimod on Immune-mediated Mechanisms of Neurodegeneration in Multiple Sclerosis - a Preclinical Study
Sponsor:
Neuromed IRCCS
Organization:
Neuromed IRCCS
Study Overview
Official Title:
Effects of Ozanimod on Immune-mediated Mechanisms of Neurodegeneration in Multiple Sclerosis - a Preclinical Study
Status:
UNKNOWN
Status Verified Date:
2022-02
Last Known Status:
NOT_YET_RECRUITING
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
This is a prospective non interventional study including patients with Relapsing-Remitting Multiple Sclerosis RRMS or with Neuromyelitis Optica Spectrum Disorders NMOSD and healthy subjects who are enrolled within the routinely programmed clinical examinations at the IRCCS Neuromed Pozzilli Italy IRCCS Polyclinic Hospital San Martino Genoa Italy and SantAndrea Hospital - University of Rome La Sapienza Rome Italy
Specifically the study investigates how ozanimod may contrast neurodegenerative mechanisms triggered by both arms of the adaptive immune response T and B cells and by their suboptimal regulation in MS Overall the project aims at assessing by in vitro experiments there will be no patients on treatment with ozanimod and the drug will be only used in vitro
AIM1 ozanimod ability to modulate the synaptotoxic effect of T-cells derived from patients with MS relapse in a MS-chimeric ex-vivo model and to identify possible mediators IRCCS Neuromed-Pozzilli in collaboration with Synaptic Immunopathology Laboratory Dep Systems Medicine Tor Vergata University of Rome AIM2 ozanimod ability to reduce the cytokine-mediated breakdown of the BBB and the migration of the here studied immune cells through ex vivo models of BBB IRCCS Polyclinic Hospital San Martino AIM3 ozanimod ability to affect the migration properties of Epstein Barr virus EBV infected B cells in MS SantAndrea Hospital AIM4 ozanimod ability to modulate the number andor function of regulatory T cells Treg a lymphocyte population playing a key role in the control of pathogenic adaptive immune responses Treg Cell Laboratory Università degli Studi di Napoli Federico II Naples Italy receiving blood samples from Neuromed Hospital and SantAndrea Hospital not recruiting unit
The work of the four labs is conceptually and operationally integrated the labs at IRCCS Neuromed-PozzilliTor Vergata University Aim1 and at Polyclinic Hospital San Martino Aim2 will investigate the effects of ozanimod on well-known mechanisms of damage in MS inflammatory synaptopathy and BBB damage and immune cell migration The lab at SantAndrea Hospital Aim3 will verify whether B cells infected by different EBV genotypes are involved in BBB migration and how ozanimod may interfere with this mechanism The Treg Cell Laboratory Aim4 will investigate whether ozanimod can also act upstream of these mechanisms by regulating the adaptive immune response
Specifically the study investigates how ozanimod may contrast neurodegenerative mechanisms triggered by both arms of the adaptive immune response T and B cells and by their suboptimal regulation in MS Overall the project aims at assessing by in vitro experiments there will be no patients on treatment with ozanimod and the drug will be only used in vitro
AIM1 ozanimod ability to modulate the synaptotoxic effect of T-cells derived from patients with MS relapse in a MS-chimeric ex-vivo model and to identify possible mediators IRCCS Neuromed-Pozzilli in collaboration with Synaptic Immunopathology Laboratory Dep Systems Medicine Tor Vergata University of Rome AIM2 ozanimod ability to reduce the cytokine-mediated breakdown of the BBB and the migration of the here studied immune cells through ex vivo models of BBB IRCCS Polyclinic Hospital San Martino AIM3 ozanimod ability to affect the migration properties of Epstein Barr virus EBV infected B cells in MS SantAndrea Hospital AIM4 ozanimod ability to modulate the number andor function of regulatory T cells Treg a lymphocyte population playing a key role in the control of pathogenic adaptive immune responses Treg Cell Laboratory Università degli Studi di Napoli Federico II Naples Italy receiving blood samples from Neuromed Hospital and SantAndrea Hospital not recruiting unit
The work of the four labs is conceptually and operationally integrated the labs at IRCCS Neuromed-PozzilliTor Vergata University Aim1 and at Polyclinic Hospital San Martino Aim2 will investigate the effects of ozanimod on well-known mechanisms of damage in MS inflammatory synaptopathy and BBB damage and immune cell migration The lab at SantAndrea Hospital Aim3 will verify whether B cells infected by different EBV genotypes are involved in BBB migration and how ozanimod may interfere with this mechanism The Treg Cell Laboratory Aim4 will investigate whether ozanimod can also act upstream of these mechanisms by regulating the adaptive immune response
Detailed Description:
Multiple sclerosis MS is a multifactorial and complex disease with several and tangled pathophysiological mechanisms responsible for the inflammatory and neurodegenerative damage Therapies targeting inflammatory pathways have brought substantial progress in the treatment of relapsing-remitting MS Disappointingly the impact of these treatments on neurodegeneration is limited To tackle the complexity of the problem combination therapies may be necessary However safety tolerability and cost issues have so far limited this option Single therapies that target at once inflammation and neurodegeneration are an appealing alternative However a comprehensive understanding of their effects is essential to maximize their impact
Ozanimod is a molecule that targets simultaneously inflammation and neurodegeneration Its impact on the latter is less well understood The drug produces its effects by acting on two S1P1 and S1P5 of the five sphingosine-1-phosphate G protein-coupled receptor subtypes While the molecular targets of the drug are precisely defined much remains to be learned about the many biological effects that may follow receptors modulation
A collaborative and integrated approach is proposed to deepen our understanding of ozanimods effects on the complex pathophysiology of MS
The research will focus on crucial aspects for MS biology Specifically it will be investigated how ozanimod may contrast neurodegenerative mechanisms triggered by both arms of the adaptive immune response T and B cells and by their suboptimal regulation in MS Overall the project aims at investigating
AIM1 ozanimod ability to modulate the synaptotoxic effect of T-cells derived from active MS patients in a MS-chimeric ex-vivo model and to identify possible mediators IRCCS Neuromed-Pozzilli in collaboration with Synaptic Immunopathology Laboratory Dep Systems Medicine Tor Vergata University of Rome Inflammatory synaptopathy namely synapse loss and synaptic dysfunction is emerging as an important hallmark of grey matter pathology in MS and its mouse model experimental autoimmune encephalomyelitis EAE In the past years evidence has been provided of a direct role of T cell derived from MS patients or EAE mice in inducing glutamate-mediated excitotoxicity a prominent form of synaptopathy detected in the MSEAE brain Notably several lines of evidence indicate that limiting glutamate-excitotoxicity is an attractive therapeutic strategy to treat MS
Recently the modulatory effects of ozanimod on inflammatory glutamate-mediated excitotoxicity was investigated and the different involvement of sphingosine receptor subtypes S1P1 and S1P5 were evaluated in the EAE model Two main results emerged first the S1P1S1P5 modulator ozanimod has central neuroprotective effects likely mediated by an action on microglia cells and infiltrating lymphocytes resulting in a reduced release of proinflammatory cytokines the main players of inflammatory synaptopathy Second the central delivery of a selective S1P1 modulator showed neuroprotective effects in terms of both EAE clinical score and inflammatory synaptopathy suggesting a primary involvement of this receptor subtype in ozanimod- induced neuroprotection also in MS
In the present project the MS chimeric ex-vivo model recently developed in Tor Vergata University lab will be used T cells isolated from active relapsing remitting MS patients but not from non-active naMS and healthy subject HS induced an increase of the spontaneous glutamatergic currents reminiscent of the alterations induced by EAE T cells
In this specific aim of the project the following aspect will be assessed i to investigate if ozanimod is able to modulate the synaptotoxic effect of T dell derived from active MS patients in the MS-chimeric ex-vivo model and ii to identify sphingosine receptor subtypes and iii soluble molecules potentially involved
A role for ozanimod in dampening excitotoxic damage caused by synaptotoxic T-cell circulating in the MS brain independently of a prevention of T cell trafficking is expected to be revealed
AIM2 ozanimod ability to reduce the cytokine-mediated breakdown of the BBB and the migration of the here studied immune cells through ex vivo models of BBB IRCCS Polyclinic Hospital San Martino B and T lymphocytes and NK cells are involved in MS pathogenesis Lassmann 2019 They differentially use S1P1 and S1P5 receptors to regulate their traffic across secondary lymphoid tissues and to enter the CNS through the BBB Sallusto et al 2012 In turn these receptors are important in maintaining the integrity and in restoring the function of BBB Chun 2021
Modulators of S1P1 and S1P5 receptors are widely used to treat MS but their mode of action is only partially known due to the complex downstream effects of the combined interactions among receptors localized at the membrane surfaces of immune cells and at the endothelial cells and podocytes of the BBB These modulating features have been mainly studied for fingolimod the first drug of this class used in MS whereas ozanimods mechanisms of action need in-depth investigations Indeed this drug has chemical structure different from that of fingolimod a binding restricted to S1P1 and S1P5 receptors vs S1P1 S1P3 S1P4 and S1P5 of fingolimod different pharmacokinetics and shorter time of elimination half-life Chun et al 2021
The purpose of this specific aim is to evaluate in ex vivo models of the human BBB exploited through transwell-based systems and organoids the effects of ozanimod on i maintaining of BBB integrity in a context of cytokine-mediated barrier breakdown ii impairing the migration of T cells and NK cells across BBB
The ex-vivo exposure of peripheral blood mononuclear cells PBMCs from MS patients to ozanimod is expected to reduce the damage of the BBB mediated by pro-inflammatory cytokines as well as to mitigate ability of these immune cells across the BBB PBMCs of patients with NMOSDs will be also used as inflammatory controls since in these disorders Th17 cells have a major pathogenetic role together with the BBB-targeting AQP-4 antibodies
AIM3 ozanimod ability to affect the migration properties of Epstein-Barr virus EBV infected B cells in MS SantAndrea Hospital
In the general population the memory subset of B lymphocytes proved to be decreased during therapy with S1P receptors modulators is the one that selectively harbors EBV as a latently infecting agent Several studies on EBV genotypes demonstrated an association of specific Epstein- Barr nuclear antigen 2 EBNA2 gene variants with MS EBNA2 acts as a transactivator on both viral and cellular gene promotersenhancers by obligatory interactions with host cell transcription factors Taking into consideration the complex interplay between EBNA2 and cellular components it is likely that EBNA2 variants may affect the processes of early infection and participate in the dysregulated virus-host interactions in MS Also MS-associated EBNA2 variants seem to interfere with B cell migration and maturation within germinal centers Despite the presence of EBV infection in the MS brain is controversial some observations support evidence of an intrathecal reactivation and virus-driven immunopathogenic response in MS The hypothesis of this specific aim is that ozanimod can have immunomodulatory effects on B cells and EBV infected cells in a MS context As already mentioned ozanimod is a potent S1P1R modulator that inhibits the egress of lymphocytes from lymphoid tissues B cells and lymphoblastoid cell lines spontaneously outgrowing spLCLs or in vitro infected with the EBV laboratory strain B958 958LCLs will be used to study how EBV infection affects migration capacity and how this capacity is affected by ozanimod
Specifically the following aspect will be investigated i the migration properties of B cells spLCLs and B958LCLs from MS patients and age- and sex-matched healthy donors HD ii if ozanimod is able to modulate the migration properties of the above cells from patients and HD
The knowledge of ozanimod influence on B cells and EBV-infected B cells migration through lymphoid organs and even in the brain will be importantly improved by this research
AIM4 ozanimod ability to modulate the number andor function of regulatory T cells Treg a lymphocyte population playing a key role in the control of autoimmune responses Treg Cell Laboratory Università degli Studi di Napoli Federico II Naples Italy receiving blood samples from Neuromed Hospital and SantAndrea Hospital not recruiting unit Intracellular metabolic pathways are able to control the induction and function of different immune cellular subsets indeed glycolysis is indispensable for the generation of human iTreg cells from Tconv cells In addition a glycolytic defect during the activation of Tconv cells in MS and T1D subjects was associated with a reduced induction and suppressive function of iTreg cells in these patients suggesting a defective function of iTreg cells during autoimmunity The effect of ozanimod on the cellular metabolic profile of T lymphocytes and of iTreg cells will be investigated In particular this specific aim will evaluate i the capacity of ozanimod to affect T cell and iTreg cell functionactivity activation proliferation suppression Foxp3 induction and ii the metabolic asset of circulating T cell populations ie measurement of glycolysis and oxidative phosphorylation purified from healthy subjects and MS patients
Ozanimod is expected to be able to control immunometabolism of Tconv and iTreg cells in RRMS patients This aim will also assess whether ozanimod is able to modify the induction of iTreg cells in healthy controls and RRMS patients
Ozanimod is a molecule that targets simultaneously inflammation and neurodegeneration Its impact on the latter is less well understood The drug produces its effects by acting on two S1P1 and S1P5 of the five sphingosine-1-phosphate G protein-coupled receptor subtypes While the molecular targets of the drug are precisely defined much remains to be learned about the many biological effects that may follow receptors modulation
A collaborative and integrated approach is proposed to deepen our understanding of ozanimods effects on the complex pathophysiology of MS
The research will focus on crucial aspects for MS biology Specifically it will be investigated how ozanimod may contrast neurodegenerative mechanisms triggered by both arms of the adaptive immune response T and B cells and by their suboptimal regulation in MS Overall the project aims at investigating
AIM1 ozanimod ability to modulate the synaptotoxic effect of T-cells derived from active MS patients in a MS-chimeric ex-vivo model and to identify possible mediators IRCCS Neuromed-Pozzilli in collaboration with Synaptic Immunopathology Laboratory Dep Systems Medicine Tor Vergata University of Rome Inflammatory synaptopathy namely synapse loss and synaptic dysfunction is emerging as an important hallmark of grey matter pathology in MS and its mouse model experimental autoimmune encephalomyelitis EAE In the past years evidence has been provided of a direct role of T cell derived from MS patients or EAE mice in inducing glutamate-mediated excitotoxicity a prominent form of synaptopathy detected in the MSEAE brain Notably several lines of evidence indicate that limiting glutamate-excitotoxicity is an attractive therapeutic strategy to treat MS
Recently the modulatory effects of ozanimod on inflammatory glutamate-mediated excitotoxicity was investigated and the different involvement of sphingosine receptor subtypes S1P1 and S1P5 were evaluated in the EAE model Two main results emerged first the S1P1S1P5 modulator ozanimod has central neuroprotective effects likely mediated by an action on microglia cells and infiltrating lymphocytes resulting in a reduced release of proinflammatory cytokines the main players of inflammatory synaptopathy Second the central delivery of a selective S1P1 modulator showed neuroprotective effects in terms of both EAE clinical score and inflammatory synaptopathy suggesting a primary involvement of this receptor subtype in ozanimod- induced neuroprotection also in MS
In the present project the MS chimeric ex-vivo model recently developed in Tor Vergata University lab will be used T cells isolated from active relapsing remitting MS patients but not from non-active naMS and healthy subject HS induced an increase of the spontaneous glutamatergic currents reminiscent of the alterations induced by EAE T cells
In this specific aim of the project the following aspect will be assessed i to investigate if ozanimod is able to modulate the synaptotoxic effect of T dell derived from active MS patients in the MS-chimeric ex-vivo model and ii to identify sphingosine receptor subtypes and iii soluble molecules potentially involved
A role for ozanimod in dampening excitotoxic damage caused by synaptotoxic T-cell circulating in the MS brain independently of a prevention of T cell trafficking is expected to be revealed
AIM2 ozanimod ability to reduce the cytokine-mediated breakdown of the BBB and the migration of the here studied immune cells through ex vivo models of BBB IRCCS Polyclinic Hospital San Martino B and T lymphocytes and NK cells are involved in MS pathogenesis Lassmann 2019 They differentially use S1P1 and S1P5 receptors to regulate their traffic across secondary lymphoid tissues and to enter the CNS through the BBB Sallusto et al 2012 In turn these receptors are important in maintaining the integrity and in restoring the function of BBB Chun 2021
Modulators of S1P1 and S1P5 receptors are widely used to treat MS but their mode of action is only partially known due to the complex downstream effects of the combined interactions among receptors localized at the membrane surfaces of immune cells and at the endothelial cells and podocytes of the BBB These modulating features have been mainly studied for fingolimod the first drug of this class used in MS whereas ozanimods mechanisms of action need in-depth investigations Indeed this drug has chemical structure different from that of fingolimod a binding restricted to S1P1 and S1P5 receptors vs S1P1 S1P3 S1P4 and S1P5 of fingolimod different pharmacokinetics and shorter time of elimination half-life Chun et al 2021
The purpose of this specific aim is to evaluate in ex vivo models of the human BBB exploited through transwell-based systems and organoids the effects of ozanimod on i maintaining of BBB integrity in a context of cytokine-mediated barrier breakdown ii impairing the migration of T cells and NK cells across BBB
The ex-vivo exposure of peripheral blood mononuclear cells PBMCs from MS patients to ozanimod is expected to reduce the damage of the BBB mediated by pro-inflammatory cytokines as well as to mitigate ability of these immune cells across the BBB PBMCs of patients with NMOSDs will be also used as inflammatory controls since in these disorders Th17 cells have a major pathogenetic role together with the BBB-targeting AQP-4 antibodies
AIM3 ozanimod ability to affect the migration properties of Epstein-Barr virus EBV infected B cells in MS SantAndrea Hospital
In the general population the memory subset of B lymphocytes proved to be decreased during therapy with S1P receptors modulators is the one that selectively harbors EBV as a latently infecting agent Several studies on EBV genotypes demonstrated an association of specific Epstein- Barr nuclear antigen 2 EBNA2 gene variants with MS EBNA2 acts as a transactivator on both viral and cellular gene promotersenhancers by obligatory interactions with host cell transcription factors Taking into consideration the complex interplay between EBNA2 and cellular components it is likely that EBNA2 variants may affect the processes of early infection and participate in the dysregulated virus-host interactions in MS Also MS-associated EBNA2 variants seem to interfere with B cell migration and maturation within germinal centers Despite the presence of EBV infection in the MS brain is controversial some observations support evidence of an intrathecal reactivation and virus-driven immunopathogenic response in MS The hypothesis of this specific aim is that ozanimod can have immunomodulatory effects on B cells and EBV infected cells in a MS context As already mentioned ozanimod is a potent S1P1R modulator that inhibits the egress of lymphocytes from lymphoid tissues B cells and lymphoblastoid cell lines spontaneously outgrowing spLCLs or in vitro infected with the EBV laboratory strain B958 958LCLs will be used to study how EBV infection affects migration capacity and how this capacity is affected by ozanimod
Specifically the following aspect will be investigated i the migration properties of B cells spLCLs and B958LCLs from MS patients and age- and sex-matched healthy donors HD ii if ozanimod is able to modulate the migration properties of the above cells from patients and HD
The knowledge of ozanimod influence on B cells and EBV-infected B cells migration through lymphoid organs and even in the brain will be importantly improved by this research
AIM4 ozanimod ability to modulate the number andor function of regulatory T cells Treg a lymphocyte population playing a key role in the control of autoimmune responses Treg Cell Laboratory Università degli Studi di Napoli Federico II Naples Italy receiving blood samples from Neuromed Hospital and SantAndrea Hospital not recruiting unit Intracellular metabolic pathways are able to control the induction and function of different immune cellular subsets indeed glycolysis is indispensable for the generation of human iTreg cells from Tconv cells In addition a glycolytic defect during the activation of Tconv cells in MS and T1D subjects was associated with a reduced induction and suppressive function of iTreg cells in these patients suggesting a defective function of iTreg cells during autoimmunity The effect of ozanimod on the cellular metabolic profile of T lymphocytes and of iTreg cells will be investigated In particular this specific aim will evaluate i the capacity of ozanimod to affect T cell and iTreg cell functionactivity activation proliferation suppression Foxp3 induction and ii the metabolic asset of circulating T cell populations ie measurement of glycolysis and oxidative phosphorylation purified from healthy subjects and MS patients
Ozanimod is expected to be able to control immunometabolism of Tconv and iTreg cells in RRMS patients This aim will also assess whether ozanimod is able to modify the induction of iTreg cells in healthy controls and RRMS patients
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None