Hypothalamic stem cells control ageing speed partly through exosomal miRNAs : Nature : Nature Research

Hypothalamic stem cells control ageing speed partly through exosomal miRNAs : Nature : Nature Research: "It has been proposed that the hypothalamus helps to control ageing, but the mechanisms responsible remain unclear. Here we develop several mouse models in which hypothalamic stem/progenitor cells that co-express Sox2 and Bmi1 are ablated, as we observed that ageing in mice started with a substantial loss of these hypothalamic cells. Each mouse model consistently displayed acceleration of ageing-like physiological changes or a shortened lifespan. Conversely, ageing retardation and lifespan extension were achieved in mid-aged mice that were locally implanted with healthy hypothalamic stem/progenitor cells that had been genetically engineered to survive in the ageing-related hypothalamic inflammatory microenvironment. Mechanistically, hypothalamic stem/progenitor cells contributed greatly to exosomal microRNAs (miRNAs) in the cerebrospinal fluid, and these exosomal miRNAs declined during ageing, whereas central treatment with healthy hypothalamic stem/progenitor cell-secreted exosomes led to the slowing of ageing. In conclusion, ageing speed is substantially controlled by hypothalamic stem cells, partially through the release of exosomal miRNAs."



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Exogenous microRNAs in maternal food pass through placenta, regulate fetal gene expression

In a new study published in the Protein & Cell, Chen-Yu

Zhang's group at Nanjing University reports that small non-coding RNAs
in maternal food can transfer through placenta to regulate fetal gene
expression.
MicroRNAs (miRNA) are a class of noncoding RNAs with lengths of
approximately 22 nucleotides that bind to target messenger RNAs to
inhibit protein translation. In previous studies, the same group has
found that plant miRNAs can enter into the host blood and tissues via
the route of food-intake. The food-derived exogenous miRNAs are
absorbed, packaged into microvesical (MV) and then secreted into
circulation by cells of animal GI tract. More importantly, once inside
the host, the food-derived exogenous miRNAs can regulate host physiology
by regulating host "target" genes in the cross-kingdom manner. In
support of this new concept, they have also found a plant microRNA,
MIR2911, which is enriched in honeysuckle, directly targets influenza A viruses (IAV) including H1N1, H5N1 and H7N9. Drinking of honeysuckle soup can prevent IAV infection and reduce H5N1-induced mice death.
Here, they report another surprising finding that exogenous plant miRNAs
and artificial synthetic small influence RNAs (siRNAs) can transfer
through the placenta and directly regulate fetus gene expression.
Firstly, exogenous plant miRNAs was detected in human umbilical cord
blood, amniotic fluid as well as animal fetuses with certain level. When
pregnant mice were administrated honeysuckle soup (the exogenous plant
microRNAs are physiological concentration in food), the plant MIR2911
was detected in fetus liver at a significant level. Finally, feeding
pregnant mice with synthetic alpha-fetoprotein (AFP, only expressed in
fetus liver) siRNA decreased significantly AFP mRNA and protein levels.
They have further demonstrated that MV- driven small RNAs are able to
pass through placenta.

Frontiers | Induction of the pro-inflammatory NF-kB-sensitive miRNA-146a by human neurotrophic viruses | Virology

A remarkably wide variety of human neurotrophic viruses—ranging from herpes simplex 1 (HSV-1;Herpesviridae; dsDNA genome) to Hantavirus (HTV; Bunyaviridae; (−)ssRNA genome) to human immunodeficiency virus (HIV; Retroviridae; (+)ssRNA genome) are associated with the rapid up-regulation of the NF-kB-sensitive pro-inflammatory microRNA-146a (miRNA-146a) in the host shortly after infection. This significant miRNA-146a up-regulation appears to be beneficial to the infecting virus as part of an immune-evasion strategy. Interestingly, miRNA-146a is also significantly up-regulated in several human central nervous system (CNS) disorders. These include Alzheimer's disease (AD) and prion disease where miRNA-146a participates in pro-inflammatory and innate-immune signaling. This opinion paper will comment on some recently clarified roles for the NF-kB-regulated, pro-inflammatory miRNA-146a in viral-induced cellular dysfunction, and how anti-miRNA-146a and/or related therapeutic strategies may be beneficial in the clinical management of a broad spectrum of viral-mediated CNS disease.

PLOS Biology: Inflammation Triggers RNA Transfer from Blood Cells to Brain Neurons

The nervous and immune systems are the two most complex systems in the
body. That complexity is amplified by the fact that each influences the
other, as they constantly exchange messages in response to environmental
and internal cues. The best known messages from immune cells are the
cytokines, which trigger changes within neurons through well-established
receptor-initiated cascades. But in a new study in this issue of PLOS Biology,
Kirsten Ridder, Stefan Momma, and colleagues show that hematopoietic
cells, which include the cells of the immune system, release messenger
RNAs (mRNAs) and microRNAs (miRNAs) that are absorbed by neurons. This
mechanism of signaling is elevated in response to inflammation, and the
transferred RNAs have the potential to influence neuronal responses.

Related articles

• Why inflammation leads to a leaky blood-brain barrier: MicroRNA-155 (medicalxpress.com)

Implication of sperm RNAs in transgenerational inheritance of the effects of early trauma in mice : Nature Neuroscience

Small non-coding RNAs (sncRNAs) are potential vectors at the interface
between genes and environment. We found that traumatic stress in early
life altered mouse microRNA (miRNA) expression, and behavioral and
metabolic responses in the progeny. Injection of sperm RNAs from
traumatized males into fertilized wild-type oocytes reproduced the
behavioral and metabolic alterations in the resulting offspring.

EpimiR: a database of curated mutual regulation between miRNAs and epigenetic modifications.

As two kinds of important transcription regulators, both epigenetic
modification and miRNA can regulate gene expression in a wide range of
complex diseases. Recently, many studies have demonstrated that
epigenetics and miRNA can regulate each other in many biological
processes. For instance, methylation of promoter-associated CpG
dinucleotides (especially in CpG islands) usually correlates with
reduced transcription levels of corresponding miRNAs and subsequently
induces the expression of miRNA target genes. Additionally, histone
modifications have been discovered to play positive or negative roles in
controlling miRNA expression in various normal cells and diseases. On
the other hand, miRNA exerts its curative effects on regulating DNA
methylation or histone modification through directly targeting
epigenetic enzymes or functional protein complexes. Thus, we developed a
comprehensive database named EpimiR to store the experimentally
validated mutual regulations between epigenetic modifications and
miRNAs. The EpimiR database have obtained 1945 regulatory relationships
between 18 types of epigenetic modification (including DNA methylation,
histone acetylation, H3K4me3 and H3K27me3, etc.) and 615 miRNAs across 6
species (including human, mouse, chicken, virus, canine, and
arabidopsis) from nearly 2000 literatures. The records that were stored
in the EpimiR database can be divided into 2 parts: Epi2miR and miR2Epi.
Users can search, submit, and download with a user-friendly interface.

Specific inhibition of diverse pathogens in human cells by synthetic microRNA-like oligonucleotides inferred from RNAi screens

Systematic genetic perturbation screening in human cells remains technically challenging. Typically, large libraries of chemically synthesized siRNA oligonucleotides are used, each designed to degrade a specific cellular mRNA via the RNA interference (RNAi) mechanism. Here, we report on data from three genome-wide siRNA screens, conducted to uncover host factors required for infection of human cells by two bacterial and one viral pathogen. We find that the majority of phenotypic effects of siRNAs are unrelated to the intended “on-target” mechanism, defined by full complementarity of the 21-nt siRNA sequence to a target mRNA. Instead, phenotypes are largely dictated by “off-target” effects resulting from partial complementarity of siRNAs to multiple mRNAs via the “seed” region (i.e., nucleotides 2–8), reminiscent of the way specificity is determined for endogenous microRNAs. Quantitative analysis enabled the prediction of seeds that strongly and specifically block infection, independent of the intended on-target effect. This prediction was confirmed experimentally by designing oligos that do not have any on-target sequence match at all, yet can strongly reproduce the predicted phenotypes. Our results suggest that published RNAi screens have primarily, and unintentionally, screened the sequence space of microRNA seeds instead of the intended on-target space of protein-coding genes. This helps to explain why previously published RNAi screens have exhibited relatively little overlap. Our analysis suggests a possible way of identifying “seed reagents” for controlling phenotypes of interest and establishes a general strategy for extracting valuable untapped information from past and future RNAi screens.

MicroRNA-132 dysregulation in Toxoplasma gondii infection has implications for dopamine signaling pathway

Congenital toxoplasmosis and toxoplasmic encephalitis can be associated
with severe neuropsychiatric symptoms. However, which host cell
processes are regulated and how Toxoplasma gondii affects these
changes remain unclear. MicroRNAs (miRNAs) are small noncoding RNA
sequences critical to neurodevelopment and adult neuronal processes by
coordinating the activity of multiple genes within biological networks.
We examined the expression of over 1000 miRNAs in human neuroepithelioma
cells in response to infection with Toxoplasma. MiR-132, a
cyclic AMP-responsive element binding (CREB)-regulated miRNA, was the
only miRNA that was substantially upregulated by all three prototype Toxoplasma strains. The increased expression of miR-132 was also documented in mice following infection with Toxoplasma.
To identify cellular pathways regulated by miR-132, we performed target
prediction followed by pathway enrichment analysis in the transcriptome
of Toxoplasma-infected mice. This led us to identify 20 genes
and dopamine receptor signaling was their strongest associated pathway.
We then examined myriad aspects of the dopamine pathway in the striatum
of Toxoplasma-infected mice 5 days after infection.
Here we report decreased expression of D1-like dopamine receptors (DRD1,
DRD5), metabolizing enzyme (MAOA) and intracellular proteins associated
with the transduction of dopamine-mediated signaling (DARPP-32
phosphorylation at Thr34 and Ser97). Increased concentrations of
dopamine and its metabolites, serotonin (5-HT) and 5-hydroxyindoleacetic
acid were documented by HPLC analysis; however, the metabolism of
dopamine was decreased and 5-HT metabolism was unchanged. Our data show
that miR-132 is upregulated following infection with Toxoplasma
and is associated with changes in dopamine receptor signaling. Our
findings provide a possible mechanism for how the parasite contributes
to the neuropathology of infection.

microRNAs are biomarkers of oncogenic human papillomavirus infections

Cellular and viral microRNAs (miRNAs) are the transcriptional products of RNA polymerase II and are regulated by transcriptional factors for their differential expression. The altered expression of miRNAs in many cancer types has been explored as a marker for possible diagnosis and therapy. We report in this study that oncogenic human papillomaviruses (HPVs) induce aberrant expression of many cellular miRNAs and that HPV18 infection produces no detectable viral miRNA. Thirteen abundant host miRNAs were specifically regulated by HPV16 and HPV18 in organotypic raft cultures of foreskin and vaginal keratinocytes as determined by miRNA array in combination with small RNA sequencing. The increase of miR-16, miR-25, miR-92a, and miR-378 and the decrease of miR-22, miR-27a, miR-29a, and miR-100 could be attributed to viral oncoprotein E6, E7, or both, all of which are known to target many cellular transcription factors. The examination of 158 cervical specimens, including 38 normal, 52 cervical intraepithelial neoplasia (CIN), and 68 cervical cancer (CC) tissues, for the expression of these eight miRNAs showed a remarkable increase of miR-25, miR-92a, and miR-378 with lesion progression but no obvious change of miR-22, miR-29a, and miR-100 among the HPV-infected tissues. Further analyses indicate that an expression ratio ≥1.5 of miR-25/92a group over miR-22/29a group could serve as a cutoff value to distinguish normal cervix from CIN and from CIN to CC.

PLOS Pathogens: How Do Viruses Avoid Inhibition by Endogenous Cellular MicroRNAs?

Upon infection of a cell, viruses encounter a wide range of miRNA species, generally more than 50 different miRNAs per cell, and these miRNAs vary greatly between tissues. For example, miR-122 is expressed at very high levels in hepatocytes, but is absent from almost all other cells, while miR-1 is primarily expressed in muscle tissue and miR-128 in neuronal cells Indeed, many of the more than 1000 known human miRNA species show a tissue-specific expression pattern, meaning that viruses that infect multiple cell types need a way to avoid inhibition by a wide range of miRNAs with distinct mRNA-targeting specificities.
A review on how they do it............

MicroRNA-23a promotes myelination in the central nervous system

Demyelinating disorders including leukodystrophies are devastating conditions that are still in need of better understanding, and both oligodendrocyte differentiation and myelin synthesis pathways are potential avenues for developing treatment. Overexpression of lamin B1 leads to leukodystrophy characterized by demyelination of the central nervous system, and microRNA-23 (miR-23) was found to suppress lamin B1 and enhance oligodendrocyte differentiation in vitro. Here, we demonstrated that miR-23a–overexpressing mice have increased myelin thickness, providing in vivo evidence that miR-23a enhances both oligodendrocyte differentiation and myelin synthesis. Using this mouse model, we explored possible miR-23atargets and revealed that the phosphatase and tensin homologue/phosphatidylinositol trisphosphate kinase/Akt/mammalian target of rapamycin pathway is modulated by miR-23a. Additionally, a long noncoding RNA, 2700046G09Rik, was identified as a miR-23a target and modulates phosphatase and tensin homologue itself in a miR-23a–dependent manner. The data presented here imply a unique role for miR-23ain the coordination of proteins and noncoding RNAs in generating and maintaining healthy myelin.

Exploring the role of human miRNAs in virus–host interactions using systematic overlap analysis

Human miRNAs have recently been found to have important roles in viral replication. Understanding the patterns and details of human miRNA interactions during virus–host interactions may help uncover novel antiviral therapies. Based on the abundance of knowledge available regarding protein–protein interactions (PPI), virus–host protein interactions, experimentally validated human miRNA-target pairs and transcriptional regulation of human miRNAs, it is possible to explore the complex regulatory network that exists between viral proteins and human miRNAs at the system level.

Results: By integrating current data regarding the virus–human interactome and human miRNA-target pairs, the overlap between targets of viral proteins and human miRNAs was identified and found to represent topologically important proteins (e.g. hubs or bottlenecks) at the global center of the human PPI network. Viral proteins and human miRNAs were also found to significantly target human PPI pairs. Furthermore, an overlap analysis of virus targets and transcription factors (TFs) of human miRNAs revealed that viral proteins preferentially target human miRNA TFs, representing a new pattern of virus–host interactions. Potential feedback loops formed by viruses, human miRNAs and miRNA TFs were also identified, and these may be exploited by viruses resulting in greater virulence and more effective replication strategies.

Exploring the role of human miRNAs in virus–host interactions using systematic overlap analysis

Human miRNAs have recently been found to have important roles in viral replication. Understanding the patterns and details of human miRNA interactions during virus–host interactions may help uncover novel antiviral therapies. Based on the abundance of knowledge available regarding protein–protein interactions (PPI), virus–host protein interactions, experimentally validated human miRNA-target pairs and transcriptional regulation of human miRNAs, it is possible to explore the complex regulatory network that exists between viral proteins and human miRNAs at the system level.

Results: By integrating current data regarding the virus–human interactome and human miRNA-target pairs, the overlap between targets of viral proteins and human miRNAs was identified and found to represent topologically important proteins (e.g. hubs or bottlenecks) at the global center of the human PPI network. Viral proteins and human miRNAs were also found to significantly target human PPI pairs. Furthermore, an overlap analysis of virus targets and transcription factors (TFs) of human miRNAs revealed that viral proteins preferentially target human miRNA TFs, representing a new pattern of virus–host interactions. Potential feedback loops formed by viruses, human miRNAs and miRNA TFs were also identified, and these may be exploited by viruses resulting in greater virulence and more effective replication strategies.

A microRNA Blood Test for Alzheimer’s? | The Scientist Magazine®

 A new test predicts Alzheimer’s disease with 93 percent accuracy, according to a study published in Genome Biology. Testing the blood of 202 people for 140 different microRNAs (miRNAs), a team of researchers at Saarland University, in Germany, identified 12 RNA fragments circulating at consistently different levels in healthy people and patients with Alzheimer’s, BBC News reported.

Neurobiology of Disease - MicroRNA-382 expression is elevated in the olfactory neuroepithelium of schizophrenia patients

Schizophrenia is a common neuropsychiatric disorder that has a strong genetic component. MicroRNAs (miRNAs) have been implicated in neurodevelopmental and psychiatric disorders including schizophrenia, as indicated by their dysregulation in post-mortem brain tissues and in peripheral blood of schizophrenia patients. The olfactory epithelium (OE) is one of the few accessible neural tissues that contain neurons and their stem cells. Previous studies showed that OE-derived tissues and cells can be safely and easily collected from live human subjects and may provide a “window” into neuronal processes involved in disorders such as schizophrenia, while avoiding the limitations of using postmortem brain samples or non-neuronal tissues. In this study, we found that the brain-enriched miR-382 (miR-382-5p) expression was elevated in in vitro cultured olfactory cells, in a cohort of seven schizophrenia patients compared with seven non-schizophrenic controls. MiR-382 elevation was further confirmed in laser-capture microdissected OE neuronal tissue (LCM-OE), enriched for mature olfactory neurons, in a cohort of 18 schizophrenia patients and 18 non-schizophrenic controls. In sharp contrast, miR-382 expression could not be detected in lymphoblastoid cell lines generated from schizophrenic or non-schizophrenic individuals. We further found that miR-382 directly regulates the expression of two genes, FGFR1 and SPRY4, which are downregulated in both the cultured olfactory cells and LCM-OE derived from schizophrenia patients. These genes are involved in the fibroblast growth factor (FGF) signaling pathway, while impairment of this pathway may underlie abnormal brain development and function associated with schizophrenia. Our data suggest that miR-382 elevation detected in patients' OE-derived samples might serve to strengthen current biomarker studies in schizophrenia. This study also illustrates the potential utility of OE-derived tissues and cells as surrogate samples for the brain.

MicroRNA-382 expression is elevated in the olfactory neuroepithelium of schizophrenia patients.

Schizophrenia is a common neuropsychiatric disorder that has a strong genetic component. MicroRNAs (miRNAs) have been implicated in neurodevelopmental and psychiatric disorders including schizophrenia, as indicated by their dysregulation in post-mortem brain tissues and in peripheral blood of schizophrenia patients. The Olfactory Epithelium (OE) is one of the few accessible neural tissues that contain neurons and their stem cells. Previous studies showed that OE-derived tissues and cells can be safely and easily collected from live human subjects and may provide a "window" into neuronal processes involved in disorders such as schizophrenia, while avoiding the limitations of using postmortem brain samples or non-neuronal tissues. In this study, we found that the brain-enriched miR-382 (miR-382-5p) expression was elevated in in vitro cultured olfactory cells, in a cohort of seven schizophrenia patients compared to seven non-schizophrenic controls. MiR-382 elevation was further confirmed in laser-capture microdissected OE neuronal tissue (LCM-OE), enriched for mature olfactory neurons, in a cohort of 18 schizophrenia patients and 18 non-schizophrenic controls. In sharp contrast, miR-382 expression could not be detected in lymphoblastoid cell lines generated from schizophrenic or non-schizophrenic individuals. We further found that miR-382 directly regulates the expression of two genes, FGFR1 and SPRY4, which are downregulated in both the cultured olfactory cells and LCM-OE derived form schizophrenia patients. These genes are involved in the Fibroblast Growth Factor (FGF) signaling pathway, while impairment of this pathway may underlie abnormal brain development and function associated with schizophrenia. Our data suggest that miR-382 elevation detected in patients' OE-derived samples might serve to strengthen current biomarker studies in schizophrenia. This study also illustrates the potential utility of OE-derived tissues and cells as surrogate samples for the brain.

Circular RNA Surprise (Blocks microRNA)| The Scientist Magazine®

Some circular RNA molecules serve as molecular “sponges,” binding to and deactivating gene modulators called microRNAs to influence gene expression, according to two papers published this week (February 27) in Nature. The findings reveal a hidden world of previously unexplored RNA molecules, and act as a reminder that there is more to RNA than simply being a messenger between DNA and the proteins it encodes. - See more at: http://www.the-scientist.com//?articles.view/articleNo/34544/title/Circular-RNA-Surprise/#sthash.7Q7htOh8.dpuf

Plasma-Based Circulating MicroRNA Biomarkers for Parkinson's Disease - Journal of Parkinson's Disease

The current “gold-standard” for Parkinson's disease (PD) diagnosis is based primarily on subjective clinical rating scales related with motor features. Molecular biomarkers that are objective and quantifiable remain attractive as clinical tools to detect PD prior to its motor onsets. Objective: Here, we aimed to identify, develop, and validate plasma-based circulating microRNA (miRNAs) as biomarkers for PD. Methods: Global miRNA expressions were acquired from a discovery set of 32 PD/32 controls using microarrays. k-Top Scoring Pairs (k-TSP) algorithm and significance analysis of microarrays (SAM) were applied to obtain comprehensive panels of PD-predictive biomarkers. TaqMan miRNA-specific real-time PCR assays were performed to validate the microarray data and to evaluate the biomarker performance using a new replication set of 42 PD/30 controls. Data was analyzed in a paired PD-control fashion. The validation set was composed of 30 PD, 5 progressive supranuclear palsy, and 4 multiple system atrophy samples from a new clinical site. Results: We identified 9 pairs of PD-predictive classifiers using k-TSP analysis and 13 most differentially-expressed miRNAs by SAM. A combination of both data sets produced a panel of PD-predictive biomarkers: k-TSP1 (miR-1826/miR-450b-3p), miR-626, and miR-505, and achieved the highest predictive power of 91% sensitivity, 100% specificity, 100% positive predicted value, and 88% negative predicted value in the replication set. However, low predictive values were shown in the validation set. Conclusions: This proof-of-concept study demonstrates the feasibility of using plasma-based circulating miRNAs as biomarkers for neurodegenerative disorders such as PD and shows the challenges of molecular biomarker research using samples from multiple clinical sites.

microRNA (miRNA) speciation in Alzheimer's disease (AD) cerebrospinal fluid (CSF) and extracellular fluid (ECF).

Human cerebrospinal fluid (CSF), produced by the choroid plexus and secreted into the brain ventricles and subarachnoid space, plays critical roles in intra-cerebral transport and the biophysical and immune protection of the brain. CSF composition provides valuable insight into soluble pathogenic bio-markers that may be diagnostic for brain disease. In these experiments we analyzed amyloid beta (Aβ) peptide and micro RNA (miRNA) abundance in CSF and in short post-mortem interval (PMI <2.1 hr) brain tissue-derived extracellular fluid (ECF) from Alzheimer's disease (AD) and age-matched control neocortex. There was a trend for decreased abundance of Aβ42 in the CSF and ECF in AD but it did not reach statistical significance (mean age ~72 yr; N=12; p~0.06, ANOVA). The most abundant nucleic acids in AD CSF and ECF were miRNAs, and their speciation and inducibility were studied further. Fluorescent miRNA-array-based analysis indicated significant increases in miRNA-9, miRNA-125b, miRNA-146a, miRNA-155 in AD CSF and ECF (N=12; p<0.01, ANOVA). Primary human neuronal-glial (HNG) cell co-cultures stressed with AD-derived ECF also displayed an up-regulation of these miRNAs, an effect that was quenched using the anti-NF-кB agents caffeic acid phenethyl ester (CAPE) or 1-fluoro-2-[2-(4-methoxy-phenyl)-ethenyl]-benzene (CAY10512). Increases in miRNAs were confirmed independently using a highly sensitive LED-Northern dot-blot assay. Several of these NF-кB-sensitive miRNAs are known to be up-regulated in AD brain, and associate with the progressive spreading of inflammatory neurodegeneration. The results indicate that miRNA-9, miRNA-125b, miRNA-146a and miRNA-155 are CSF- and ECF-abundant, NF-кB-sensitive pro-inflammatory miRNAs, and their enrichment in circulating CSF and ECF suggest that they may be involved in the modulation or proliferation of miRNA-triggered pathogenic signaling throughout the brain and central nervous system (CNS).

Incorporating Information of microRNAs into Pathway Analysis in a Genome-Wide Association Study of Bipolar Disorder.

MicroRNAs (miRNAs) are known to be important post-transcriptional regulators that are involved in the etiology of complex psychiatric traits. The present study aimed to incorporate miRNAs information into pathway analysis using a genome-wide association dataset to identify relevant biological pathways for bipolar disorder (BPD). We selected psychiatric- and neurological-associated miRNAs (N = 157) from PhenomiR database. The miRNA target genes (miTG) predictions were obtained from microRNA.org. Canonical pathways (N = 4,051) were downloaded from the Molecule Signature Database. We employed a novel weighting scheme for miTGs in pathway analysis using methods of gene set enrichment analysis and sum-statistic. Under four statistical scenarios, 38 significantly enriched pathways (P-value < 0.01 after multiple testing correction) were identified for the risk of developing BPD, including pathways of ion channels associated (e.g., gated channel activity, ion transmembrane transporter activity, and ion channel activity) and nervous related biological processes (e.g., nervous system development, cytoskeleton, and neuroactive ligand receptor interaction). Among them, 19 were identified only when the weighting scheme was applied. Many miRNA-targeted genes were functionally related to ion channels, collagen, and axonal growth and guidance that have been suggested to be associated with BPD previously. Some of these genes are linked to the regulation of miRNA machinery in the literature. Our findings provide support for the potential involvement of miRNAs in the psychopathology of BPD. Further investigations to elucidate the functions and mechanisms of identified candidate pathways are needed.