Buffy Coat Transcriptomic Analysis Reveals Alterations in Host Cell Protein Synthesis and Cell Cycle in Severe COVID-19 Patients.

Transcriptome studies have reported the dysregulation of cell cycle-related genes and the global inhibition of host mRNA translation in COVID-19 cases. However, the key genes and cellular mechanisms that are most affected by the severe outcome of this disease remain unclear. For this work, the RNA-seq approach was used to study the differential expression in buffy coat cells of two groups of people infected with SARS-CoV-2: (a) Mild, with mild symptoms; and (b) SARS (Severe Acute Respiratory Syndrome), who were admitted to the intensive care unit with the severe COVID-19 outcome. Transcriptomic analysis revealed 1009 up-regulated and 501 down-regulated genes in the SARS group, with 10% of both being composed of long non-coding RNA. Ribosome and cell cycle pathways were enriched among down-regulated genes. The most connected proteins among the differentially expressed genes involved transport dysregulation, proteasome degradation, interferon response, cytokinesis failure, and host translation inhibition. Furthermore, interactome analysis showed Fibrillarin to be one of the key genes affected by SARS-CoV-2. This protein interacts directly with the N protein and long non-coding RNAs affecting transcription, translation, and ribosomal processes. This work reveals a group of dysregulated processes, including translation and cell cycle, as key pathways altered in severe COVID-19 outcomes.

Resveratrol Treatment of Autism Spectrum Disorder-A Pilot Study.

OBJECTIVES: Considering autism spectrum disorder (ASD) as a neurodevelopmental condition associated with immune system impairments, we aimed to evaluate the potential benefits, efficacy, tolerability, and safety of the anti-inflammatory, antioxidant, and neuroprotective trans -resveratrol (RSV) in behavioral impairments and in a set of 8 microRNAs (miR) related to the immune system in pediatric subjects with ASD. METHODS: This is an open-label pilot trial over a 3 months (90 days) study follow-up period designed to assess the effect of 200 mg/d RSV on 5 boys aged 10 to 13 (11.8 ± 1.1) years diagnosed with ASD according to Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition . RESULTS: The RSV treatment significantly reduced the Aberrant Behavior Checklist total score ( P = 0.042) and Irritability ( P = 0.041), with no alteration in Stereotypical Behavior ( P = 0.066), Hyperactivity ( P = 0.068), and Lethargy/Social Withdrawal ( P = 0.078) subscales. On the Clinical Global Impression scale, 3 individuals showed marked improvement in behavior; one showed mild improvement, and the other had no changes. The RSV treatment increased the miR-195-5p ( P = 0.043), an important modulator of targets related to inflammatory and immunological pathways. RSV administration did not present adverse effects and did not alter clinical laboratory results. CONCLUSIONS: RSV is a safe molecule for administrating in the pediatric population, able to modulate behavior alterations and molecules associated with the immune system, becoming a promising therapeutic strategy for large-scale studies in ASD, to investigate both behavioral and molecular approaches.

Zika virus disrupts gene expression in human myoblasts and myotubes: Relationship with susceptibility to infection.

The tropism of Zika virus (ZIKV) has been described in the nervous system, blood, placenta, thymus, and skeletal muscle. We investigated the mechanisms of skeletal muscle susceptibility to ZIKV using an in vitro model of human skeletal muscle myogenesis, in which myoblasts differentiate into myotubes. Myoblasts were permissive to ZIKV infection, generating productive viral particles, while myotubes controlled ZIKV replication. To investigate the underlying mechanisms, we used gene expression profiling. First, we assessed gene changes in myotubes compared with myoblasts in the model without infection. As expected, we observed an increase in genes and pathways related to the contractile muscle system in the myotubes, a reduction in processes linked to proliferation, migration and cytokine production, among others, confirming the myogenic capacity of our system in vitro. A comparison between non-infected and infected myoblasts revealed more than 500 differentially expressed genes (DEGs). In contrast, infected myotubes showed almost 2,000 DEGs, among which we detected genes and pathways highly or exclusively expressed in myotubes, including those related to antiviral and innate immune responses. Such gene modulation could explain our findings showing that ZIKV also invades myotubes but does not replicate in these differentiated cells. In conclusion, we showed that ZIKV largely (but differentially) disrupts gene expression in human myoblasts and myotubes. Identifying genes involved in myotube resistance can shed light on potential antiviral mechanisms against ZIKV infection.

HDAC inhibitor affects soybean miRNA482bd expression under salt and osmotic stress.

MicroRNAs (miRNAs) are small non-coding molecules that modulate gene expression through targeting mRNA by specific-sequence cleavage, translation inhibition, or transcriptional regulation. miRNAs are key molecules in regulatory networks in abiotic stresses such as salt stress and water deficit in plants. Throughout the world, soybean is a critical crop, the production of which is affected by environmental stress conditions. In this study, RNA-Seq libraries from leaves of soybean under salt treatment were analyzed. 17 miRNAs and 31 putative target genes were identified with inverse differential expression patterns, indicating miRNA-target interaction. The differential expression of six miRNAs, including miR482bd-5p, and their potential targets, were confirmed by RT-qPCR. The miR482bd-5p expression was repressed, while its potential HEC1 and BAK1 targets were increased. Polyethylene glycol experiment was used to simulate drought stress, and miR482bd-5p, HEC1, and BAK1 presented a similar expression pattern, as found in salt stress. Histone modifications occur in response to abiotic stress, where histone deacetylases (HDACs) can lead to gene repression and silencing. The miR482bd-5p epigenetic regulation by histone deacetylation was evaluated by using the SAHA-HDAC inhibitor. The miR482bd-5p was up-regulated, and HEC1 was down-regulated under SAHA-salt treatment. It suggests an epigenetic regulation, where the miRNA gene is repressed by HDAC under salt stress, reducing its transcription, with an associated increase in the HEC1 target expression.

Circular and Micro RNAs from Arabidopsis thaliana Flowers Are Simultaneously Isolated from AGO-IP Libraries.

Competing endogenous RNAs (ceRNAs) are natural transcripts that can act as endogenous sponges of microRNAs (miRNAs), modulating miRNA action upon target mRNAs. Circular RNAs (circRNAs) are one among the various classes of ceRNAs. They are produced from a process called back-splicing and have been identified in many eukaryotes. In plants, their effective action as a miRNA sponge was not yet demonstrated. To address this question, public mRNAseq data from Argonaute-immunoprecipitation libraries (AGO-IP) of Arabidopsis thaliana flowers were used in association with a bioinformatics comparative multi-method to identify putative circular RNAs. A total of 27,812 circRNAs, with at least two reads at the back-splicing junction, were identified. Further analyses were used to select those circRNAs with potential miRNAs binding sites. As AGO forms a ternary complex with miRNA and target mRNA, targets count in AGO-IP and input libraries were compared, demonstrating that mRNA targets of these miRNAs are enriched in AGO-IP libraries. Through this work, five circRNAs that may function as miRNA sponges were identified and one of them were validated by PCR and sequencing. Our findings indicate that this post-transcriptional regulation can also occur in plants.

Circular RNAs and Plant Stress Responses.

Circular RNAs (circRNAs) are a novel class of noncoding RNAs that have been extensively explored in the past few years. The advent of new high-throughput sequencing technologies coupled with bioinformatics tools revealed the presence of these molecules in the transcriptome of a wide range of organisms. In animals, circRNAs can modulate gene expression and act as sponges of miRNAs to inhibit their activity. It has been demonstrated that they have the potential to be diagnostic biomarkers as their expression is closely associated to human diseases, such as Alzheimer and cancer. However, in plants their function remains elusive. Recently, the role of the circRNAs in plant stress responses has been studied. During the infection of Pseudomonas syringae in kiwifruit plants, 584 circRNAs were differentially expressed in leaf samples, and a group of them could be further associated with the stage of infection. Under phosphate deficiency conditions, 27 rice circRNAs were reported to be differentially expressed. In tomato, 163 circRNAs demonstrated chilling-responsive expression, with 102 containing miRNA-binding sites and are predicted to act as miRNA sponges. Additionally, Arabidopsis seedlings presented 1583 heat-specific circRNAs, and it was also reported that heat stress could increase the quantity, length, and alternative circularization events of circRNAs. Finally, wheat seedlings under dehydration stress had 62 circRNAs differentially expressed, with 6 being predicted as miRNA sponges. Although the role of plant circRNAs during the biotic and abiotic stresses is still poorly characterised, these molecules have the potential to expand the number of targets and tools in the biotechnology field.

Unusual RNA plant virus integration in the soybean genome leads to the production of small RNAs.

Horizontal gene transfer (HGT) is known to be a major force in genome evolution. The acquisition of genes from viruses by eukaryotic genomes is a well-studied example of HGT, including rare cases of non-retroviral RNA virus integration. The present study describes the integration of cucumber mosaic virus RNA-1 into soybean genome. After an initial metatranscriptomic analysis of small RNAs derived from soybean, the de novo assembly resulted a 3029-nt contig homologous to RNA-1. The integration of this sequence in the soybean genome was confirmed by DNA deep sequencing. The locus where the integration occurred harbors the full RNA-1 sequence followed by the partial sequence of an endogenous mRNA and another sequence of RNA-1 as an inverted repeat and allowing the formation of a hairpin structure. This region recombined into a retrotransposon located inside an exon of a soybean gene. The nucleotide similarity of the integrated sequence compared to other Cucumber mosaic virus sequences indicates that the integration event occurred recently. We described a rare event of non-retroviral RNA virus integration in soybean that leads to the production of a double-stranded RNA in a similar fashion to virus resistance RNAi plants.

Metatranscriptomic analysis of small RNAs present in soybean deep sequencing libraries.

A large number of small RNAs unrelated to the soybean genome were identified after deep sequencing of soybean small RNA libraries. A metatranscriptomic analysis was carried out to identify the origin of these sequences. Comparative analyses of small interference RNAs (siRNAs) present in samples collected in open areas corresponding to soybean field plantations and samples from soybean cultivated in greenhouses under a controlled environment were made. Different pathogenic, symbiotic and free-living organisms were identified from samples of both growth systems. They included viruses, bacteria and different groups of fungi. This approach can be useful not only to identify potentially unknown pathogens and pests, but also to understand the relations that soybean plants establish with microorganisms that may affect, directly or indirectly, plant health and crop production.