Dysregulation of MicroRNAs and Target Genes Networks in Peripheral Blood of Patients With Sporadic Amyotrophic Lateral Sclerosis

Abstract

Amyotrophic lateral sclerosis (ALS) is a progressive and fatal neurodegenerative disease. While genetics and other factors contribute to ALS pathogenesis, critical knowledge is still missing and validated biomarkers for monitoring the disease activity have not yet been identified. To address those aspects we carried out this study with the primary aim of identifying possible miRNAs/mRNAs dysregulation associated with the sporadic form of the disease (sALS). Additionally, we explored miRNAs as modulating factors of the observed clinical features. Study included 56 sALS and 20 healthy controls (HCs). We analyzed the peripheral blood samples of sALS patients and HCs with a high-throughput next-generation sequencing followed by an integrated bioinformatics/biostatistics analysis. Results showed that 38 miRNAs (let-7a-5p, let-7d-5p, let-7f-5p, let-7g-5p, let-7i-5p, miR-103a-3p, miR-106b-3p, miR-128-3p, miR-130a-3p, miR-130b-3p, miR-144-5p, miR-148a-3p, miR-148b-3p, miR-15a-5p, miR-15b-5p, miR-151a-5p, miR-151b, miR-16-5p, miR-182-5p, miR-183-5p, miR-186-5p, miR-22-3p, miR-221-3p, miR-223-3p, miR-23a-3p, miR-26a-5p, miR-26b-5p, miR-27b-3p, miR-28-3p, miR-30b-5p, miR-30c-5p, miR-342-3p, miR-425-5p, miR-451a, miR-532-5p, miR-550a-3p, miR-584-5p, miR-93-5p) were significantly downregulated in sALS. We also found that different miRNAs profiles characterized the bulbar/spinal onset and the progression rate. This observation supports the hypothesis that miRNAs may impact the phenotypic expression of the disease. Genes known to be associated with ALS (e.g., PARK7, C9orf72, ALS2, MATR3, SPG11, ATXN2) were confirmed to be dysregulated in our study. We also identified other potential candidate genes like LGALS3 (implicated in neuroinflammation) and PRKCD (activated in mitochondrial-induced apoptosis). Some of the downregulated genes are involved in molecular bindings to ions (i.e., metals, zinc, magnesium) and in ions-related functions. The genes that we found upregulated were involved in the immune response, oxidation–reduction, and apoptosis. These findings may have important implication for the monitoring, e.g., of sALS progression and therefore represent a significant advance in the elucidation of the disease’s underlying molecular mechanisms. The extensive multidisciplinary approach we applied in this study was critically important for its success, especially in complex disorders such as sALS, wherein access to genetic background is a major limitation.

Local and Systemic Humoral Response to Autologous Lineage-Negative Cells Intrathecal Administration in ALS Patients

“…Haramati et al reported that this micro-RNA is down-regulated in the genetic model of spinal muscular atrophy, which suggests its role in the development of neurodegeneration [75]; therefore, its increase in the environment of CNS may have beneficial influence on neural cells. Using high-throughput next-generation sequencing, Liguori et al showed that miR-let-7f-5p is down-regulated in peripheral blood of sALS patients [69]. Similar findings were reported by Takahashi et al, this time after analysis of ALS patients' plasma samples.…”

“…In our previous study, we found consistent expression of several tested miRNAs in CSF and plasma of ALS patients [23]. miRNA-16-5p downregulation was shown in ALS patients compared to controls [68,69] and in a murine model of early-onset Alzheimer's disease (AD) [39]. Liu et al suggested a neuroprotective role of miRNA-16-5p since, upon administration of miR-16-5p into the brains of AD mice, amyloid precursor protein levels were reduced [39].…”

Seeking a standardized normalization method for the quantification of microRNA expression

“…We thank Madadi and Soleimani for having cited our study; however, we respectfully disagree with their conclusion, which asserts that the customized miRNA normalization panel used by Raheja et al . may not be suitable, as miRNA normalizers showed dysregulated expression in other studies.…”

“…In our cited study, we investigated the miRNA expression profile in blood samples of ALS patients using a high‐throughput next‐generation sequencing (HT‐NGS) approach, followed by quantititative polymerase chain reaction (qPCR) validation. Because there are no published studies that systematically evaluated reference miRNAs to normalize blood‐derived miRNAs in the setting of ALS, 5 candidate endogenous reference miRNAs were selected from previous qPCR blood studies and from suggested miRNA endogenous controls in the TaqMan Advanced miRNA Assays white paper (Applied Biosystems, Thermo Fisher Scientific).…”

Association between miRNAs expression and cognitive performances of Pediatric Multiple Sclerosis patients: A pilot study

“…The statistical significance was determined using the one-tailed Fisher's exact test followed by the Benjamini correction; adjusted p-value <0.05 was set as the threshold value. Details of methods and software used for the analysis have been already published (Liguori, Nuzziello, Introna, et al, 2018a;Liguori, Nuzziello, Licciulli, et al, 2018b).…”