Targeted Stage-Specific Inflammatory microRNA Profiling in Urine During Disease Progression in Experimental Autoimmune Encephalomyelitis: Markers of Disease Progression and Drug Response

Abstract

Recently, microRNAs (miRNAs) have been implicated in regulating neuroinflammatory and demyelinative responses in multiple sclerosis (MS) and its mouse model of experimental autoimmune encephalomyelitis (EAE). miRNAs have also been studied as biomarkers of disease pathology and drug-response in MS. However, no complete miRNA profiling at various stages of EAE disease has been examined, especially in the urine. We carried out a systematic analysis of miRNAs in the urine exosomes as well as in the plasma and spinal cord at pre-onset, onset and peak stages of EAE established in the chronic B6 mice model. For the first time, we provide evidence that urine exosomes can be a specific and sensitive source of miRNA biomarkers for all 3 stages of EAE disease. In a significant observation, we observed that miR-155-5p expression increased in urine exosomes, plasma and spinal cord 6 days before the onset of disease, suggesting its early involvement in the pathology of EAE disease. We also analyzed the effect of Glatiramer acetate (GA; copaxone) treatment, an approved treatment for MS patients, in modulating miRNA expression at the peak of EAE disease. We identified miR-155-5p, miR-27a-3p, miR-9-5p and miR-350-5p as putative GA-treatment responsive miRNA biomarkers. Since, EAE is a mainly CD4 cells mediated disease, we also examined the above set of miRNAs and found to be significantly altered in T cells polarized to Th1 and Th17 phenotype, similar to urine exosomes. Thus, urine exosome miRNAs hold the potential to be defined as novel accessible stage-specific biomarkers of EAE (MS) disease as well as treatment response.

Neuronal microRNA regulation in Experimental Autoimmune Encephalomyelitis

“…8 0 . 0041 1.1 0.9369 downregulated in both MS active and inactive lesions versus normal brain WM 17 ; upregulated in MS CD4+ T cells versus HC 41 ; upregulated in plasma of EAE mice vs control 36 miR-142a-5p −1.1 0.8945 2 . 5 0 .…”

“…Next, we asked if the miRNAs identified by miR-Seq and validated in the neurons of EAE mice were previously described as differentially regulated in other tissues and cells in EAE and/or MS. Of the 27 regulated miRNAs in motor neurons, 13 have been previously implicated in EAE and/or MS (Table 1 ). Regulation of individual miRNAs was previously identified in immune populations including lymph nodes 29 , whole blood 30 , 31 , serum 32 – 35 , plasma 36 , peripheral blood mononuclear cells (PBMCs) 35 , 37 , leukocytes 38 , 39 , CD3+ T cells 40 , CD4+ T cells 41 , 42 , T regulatory cells (T regs ) 20 , or B cells 43 ; or in the brain within MS lesions 17 , 44 or normal appearing white matter (NAWM) 21 , 45 , 46 . In a small number of cases the miRNA of interest was genetically manipulated for investigation in EAE 47 – 50 .…”

Precision Medicine in Neurodegenerative Diseases: Some Promising Tips Coming from the microRNAs’ World

“…In addition, in the animal model of MS (experimental autoimmune encephalomyelitis, EAE), genetic deletion of the miRNA cluster miR-106a∼363 (containing Natalizumab-regulated miR-20b) resulted in a more severe disease course, and it upregulated in vivo the miR-20b target genes [89]. In addition, the treatment with Natalizumab restored the expression levels of miR-26a and miR-155 [90], which were upregulated in PBMCs of MS patients, as well as in urine exosome, plasma, and in the spinal cord samples from EAE mice [82]. Notably, miR-155 plays a central role in many processes involved in the pathogenesis of MS, such as immune cell activation, neurodegeneration and permeabilization of the BBB [18].…”

“…The pioneer study on the miRNA modulation by GA reported that the administration of GA was able to restore normal levels of miR-146a and miR-142-3p, which were upregulated in PBMCs of the RRMS patients [81]. Later, Singh et al [82] demonstrated that GA treatment was able to modulate the expression levels of miR-155-5p, miR-27a-3p, miR-9-5p, and miR-350-5p, in plasma and urine exosome, so they were suggested as potential biomarkers of drug response. Glatiramer Acetate, Interferon-β, and Dimethyl fumarate, are shown in green, blue, purple, orange, and brown, respectively.…”

The roles of extracellular vesicle microRNAs in the central nervous system

“…Recently, significant advances have been made in the search for biomarkers in MS. In EAE, differential miRNA profiles for each phase of disease (pre‐onset, onset, disease peak) may provide valuable insight into putative biomarkers for MS diagnosis (Singh et al, ). Several miRNAs identified were also differentially regulated with glatiramer acetate treated individuals, suggesting that changes in EV‐derived miRNAs may present a useful and non‐invasive method to monitor treatment efficacy.…”