Contributions
Abstract: P583
Type: Poster Sessions
Abstract Category: Therapy - Neuroprotection and Repair
Introduction: Multiple sclerosis (MS) is a central nervous system (CNS) disease in which myelin is damage by an autoimmune attack. Although there are several treatments to modulate the immune system, remyelination promotors are not available. The use of microRNAs and more concretely hsa-miR-219, has been proposed as oligodendrocyte precursor cell differentiation mediator and therefore as a remyelination inductor. Nevertheless, the administration of the microRNA to the CNS is a tricky question and delivery systems, such as the use of exosomes are under study. Exosomes are 100 nm particles released by cells and implicated in the intercellular communication which carry proteins and genetic material.
Aims: The goal of this work is to address the ability of miR-219 enriched exosomes (Ex-219) to promote OPC differentiation in vitro and to induce remyelination in an animal model of MS, the experimental autoimmune encephalomyelitis (EAE).
Methods: Exosomes were isolated from HEK 293T cells which were previously infected with the pLKO1-miR219 plasmid. Exosomes were isolated with tangential flow filtration system and sequential centrifugation steps. Exosomes were characterized by Nanotrack Particle Analysis and Cryo-TEM. Droplet Digital PCR was used to quantify the cargo of miR-219 in the exosomes. Uptake and differentiation studies were performed in OPC primary cultures obtained from P2 C57BL/6 mice and analysed by confocal microscopy and qPCR respectively. MOG35-55 EAE induced model was used to determine the ability of exosomes to decrease clinical symptoms after intranasal administration of Ex-219. T11 Nuclear Magnetic Resonance was performed to characterize the EAE lesions. Animal procedures were approved by the pertinent ethical committee.
Results: Ex-219 overexpressed miR-219 with a FC=108 when compared to non-enriched exosomes (NE-Ex). OPC were able to up-take exosomes with a 63% of efficiency. Ex-219 increased the expression of myelin related genes in OPCs (MBP, PLP1, MOG and CNPase; FC>2) and decrease the EAE score after intranasal administration (AUC p< 0.05) when compared to NE-Ex and non-treated animals.
Conclusions: To conclude, Ex-219 are able to induce OPC differentiation in vitro and to decrease EAE score after intranasal administration. In addition, exosomes have shown to be a proper microRNA delivery system for CNS diseases. These results open a promising and feasible remyelination therapy for MS and other neurodegenerative diseases.
Disclosure: Iñaki Osorio-Querejeta: nothing to disclosure
Ander Eguimendia: nothing to disclosure
Laura Moles: nothing to disclosure
Leire Iparraguirre: nothing to disclosure
Susana Carrejal-Romero: nothing to disclosure
Irantzu Ilarena: nothing to disclosure
Leslie Nash: nothing to disclosure
Lucía Sepulpeva: nothing to disclosure
Carlos San José: nothing to disclosure
Matthew Wood: nothing to disclosure
Imra Mäger: nothing to disclosure
Pedro Ramos-Cabrer: nothing to disclosure
Maider Muñoz-Culla: nothing to disclosure
David Otaegui: nothing to disclosure
Fundings: IOQ, AA and LI are founded by the Predoctoral program of the Basque Government. IOQ was also founded by EMBO Short Term Fellowships. The project was funded by Instituto de Salud Carlos III (DTS15/00069 and PI14/0093) and partially by Industry Department of the Basque Country (ElKARTEK-bmG-2016)
Abstract: P583
Type: Poster Sessions
Abstract Category: Therapy - Neuroprotection and Repair
Introduction: Multiple sclerosis (MS) is a central nervous system (CNS) disease in which myelin is damage by an autoimmune attack. Although there are several treatments to modulate the immune system, remyelination promotors are not available. The use of microRNAs and more concretely hsa-miR-219, has been proposed as oligodendrocyte precursor cell differentiation mediator and therefore as a remyelination inductor. Nevertheless, the administration of the microRNA to the CNS is a tricky question and delivery systems, such as the use of exosomes are under study. Exosomes are 100 nm particles released by cells and implicated in the intercellular communication which carry proteins and genetic material.
Aims: The goal of this work is to address the ability of miR-219 enriched exosomes (Ex-219) to promote OPC differentiation in vitro and to induce remyelination in an animal model of MS, the experimental autoimmune encephalomyelitis (EAE).
Methods: Exosomes were isolated from HEK 293T cells which were previously infected with the pLKO1-miR219 plasmid. Exosomes were isolated with tangential flow filtration system and sequential centrifugation steps. Exosomes were characterized by Nanotrack Particle Analysis and Cryo-TEM. Droplet Digital PCR was used to quantify the cargo of miR-219 in the exosomes. Uptake and differentiation studies were performed in OPC primary cultures obtained from P2 C57BL/6 mice and analysed by confocal microscopy and qPCR respectively. MOG35-55 EAE induced model was used to determine the ability of exosomes to decrease clinical symptoms after intranasal administration of Ex-219. T11 Nuclear Magnetic Resonance was performed to characterize the EAE lesions. Animal procedures were approved by the pertinent ethical committee.
Results: Ex-219 overexpressed miR-219 with a FC=108 when compared to non-enriched exosomes (NE-Ex). OPC were able to up-take exosomes with a 63% of efficiency. Ex-219 increased the expression of myelin related genes in OPCs (MBP, PLP1, MOG and CNPase; FC>2) and decrease the EAE score after intranasal administration (AUC p< 0.05) when compared to NE-Ex and non-treated animals.
Conclusions: To conclude, Ex-219 are able to induce OPC differentiation in vitro and to decrease EAE score after intranasal administration. In addition, exosomes have shown to be a proper microRNA delivery system for CNS diseases. These results open a promising and feasible remyelination therapy for MS and other neurodegenerative diseases.
Disclosure: Iñaki Osorio-Querejeta: nothing to disclosure
Ander Eguimendia: nothing to disclosure
Laura Moles: nothing to disclosure
Leire Iparraguirre: nothing to disclosure
Susana Carrejal-Romero: nothing to disclosure
Irantzu Ilarena: nothing to disclosure
Leslie Nash: nothing to disclosure
Lucía Sepulpeva: nothing to disclosure
Carlos San José: nothing to disclosure
Matthew Wood: nothing to disclosure
Imra Mäger: nothing to disclosure
Pedro Ramos-Cabrer: nothing to disclosure
Maider Muñoz-Culla: nothing to disclosure
David Otaegui: nothing to disclosure
Fundings: IOQ, AA and LI are founded by the Predoctoral program of the Basque Government. IOQ was also founded by EMBO Short Term Fellowships. The project was funded by Instituto de Salud Carlos III (DTS15/00069 and PI14/0093) and partially by Industry Department of the Basque Country (ElKARTEK-bmG-2016)