Changes In miRNA Profiles Research Articles

Pilot study on the impact of HIFU treatment on miRNA profiles in vaginal secretions of uterine fibroids and adenomyosis patients

Background High intensity focused ultrasound (HIFU) ablation treatment for uterine fibroids and adenomyosis has been long developed. The aim of this study is to investigate miRNA profile changes in vaginal secretions after HIFU treatment and their clinical relevance. Methods We prospectively collected vaginal secretions samples from 8 patients (1 with adenomyosis and 7 with fibroids) before and after HIFU treatment. RNA was isolated and miRNA profiles were analyzed using next-generation sequencing (NGS) sequencing. Results Our study showed miRNA profile change in vaginal secretion samples after HIFU treatment for uterine fibroids/adenomyosis, with 33 miRNAs upregulated and 6 downregulated overall. In fibroid cases, 31 miRNAs were upregulated and 7 downregulated, while in adenomyosis case, 41 miRNAs were upregulated and 71 downregulated. Four miRNAs (hsa-miR-7977, hsa-miR-155-5p, hsa-miR-191-5p, hsa-miR-223-3p) showed significant differences after HIFU treatment in fibroid cases, except in case 5 with the lowest treatment sonications (425 sonications) and energy input (170000 J). hsa-miR-7977 consistently showed downregulation after HIFU treatment. hsa-miR-155-5p were downregulated in case 4 with lowest treatment efficiency (2439.64 J/cm3), while they were upregulated in other cases. hsa-miR-191-5p and hsa-miR-223-3p were downregulated in cases 4 and 7, with case 7 influenced by high sonication and energy due to multiple fibroids. Conclusions HIFU treatment altered miRNA profiles in fibroids/adenomyosis patients. Notably, hsa-miR-7977, hsa-miR-155-5p, hsa-miR-191-5p, and hsa-miR-223-3p showed significant changes in fibroid cases, except in low-energy treatments. hsa-miR-7977 consistently decreased post-treatment, while hsa-miR-155-5p decreased in the least efficient cases. Further research is needed for validation.

Amniotic fluid-derived small extracellular vesicles for predicting postnatal severe outcome of congenital diaphragmatic hernia.

Congenital diaphragmatic hernia (CDH) is a life-threatening condition with high morbidity and mortality rates. The survival rate of neonates with severe CDH is reportedly only 10%-15%. However, prenatal prediction of severe cases is difficult, and the discovery of new predictive markers is an urgent issue. In this study, we focused on microRNAs (miRNAs) in amniotic fluid-derived small EVs (AF-sEVs). We identified four miRNAs (hsa-miR-127-3p, hsa-miR-363-3p, hsa-miR-493-5p, and hsa-miR-615-3p) with AUC>0.8 to classify good prognosis group and poor prognosis group in human study. The AUC for hsa-miR-127-3p and hsa-miR-615-3p, for predicting the poor prognosis, were 0.93 and 0.91, respectively. In addition, in the in vivo study, the miRNA profiles of the lung tissues of CDH rats were different from those of control rats. Additionally, two elevated miRNAs (rno-miR-215-5p and rno-miR-148a-3p) in the lung tissues of CDH rats were increased in the AF-sEVs of CDH rats. Our results suggest that severe CDH neonates can be predicted prenatally with high accuracy using miRNAs contained in AF-sEVs. Furthermore, miRNA profile changes in AF-sEVs reflected the lung status in CDH. Our findings may contribute to the development of advanced perinatal care for patients with CDH.

MicroRNA Profiles in Hematopoietic Stem Cell Transplant Recipients.

Background: Hematopoietic Stem Cell Transplant (HCT) is a potentially curative treatment for hematologic malignancies, including multiple myeloma. Biomarker investigation can guide identification of HCT recipients at-risk for poor outcomes. MicroRNAs (miRNAs) are a class of non-coding RNAs involved in the modulation and regulation of pathological processes and are emerging as prognostic and predictive biomarkers for multiple health conditions. This pilot study aimed to examine miRNA profiles associated with HCT-related risk factors and outcomes in patients undergoing autologous HCT. Methods: Patients eligible for autologous HCT were recruited and blood samples and HCT-related variables were collected. Differential expression analysis of miRNA was conducted on 24 patient samples to compare changes in miRNA profile in HCT eligible patients before and after transplant. Results: Unsupervised clustering of differentially expressed (p < .05) miRNAs pre- and post- HCT identified clusters of up- and down-regulated miRNAs. Four miRNAs (miR-125a-5p, miR-99b-5p, miR-382-5p, miR-145-5p) involved in hematopoiesis (differentiation of progenitor cells, granulocyte function, thrombopoiesis, and tumor suppression) were significantly downregulated post-HCT. Correlation analyses identified select miRNAs associated with risk factors (such as frailty, fatigue, cognitive decline) and quality of life pre- and post-HCT. Select miRNAs were correlated with platelet engraftment. Conclusion: Future studies should examine miRNA signatures in larger cohorts in association with HCT outcomes; and expand investigations in patients receiving allogeneic transplants. This will lead to identification of biomarkers for risk stratification of HCT recipients.

Characterization and profiling of the microRNA in small extracellular vesicles isolated from goat milk samples collected during the first week postpartum

Colostrum contains nutrients, immunoglobulins, and various bioactive compounds such as microRNA (miRNA). Less is known about the temporal changes in miRNA profiles in ruminant milk samples during the first week postpartum. In this study, we characterized and compared the profiles of miRNA in the small extracellular vesicles (sEV) isolated from colostrum (CM, collected immediately after parturition, n = 8) and transition milk (TM, collected 7 d postpartum, n = 8) from eight 1-yr-old Guanzhong dairy goats with a milk yield of approximately 500 kg/year. A total of 192 unique sEV-associated miRNA (transcripts per million >1 at least 4 samples in either CM or TM) were identified in all samples. There were 29 miRNA uniquely identified in the TM samples while no miRNA was uniquely identified in the CM samples. The abundance of the top 10 miRNA accounted for 82.4% ± 4.0% (± SD) of the total abundance, with let-7 families (e.g., let-7a/b/c-5p) being predominant in all samples. The top 10 miRNA were predicted to target 1,008 unique genes that may regulate pathways such as focal adhesion, TGF-β signaling, and axon guidance. The expression patterns of EV miRNA were similar between the 2 sample groups, although the abundance of let-7c-5p and miR-30a-3p was higher, whereas that of let-7i-5p and miR-103–3p was lower in CM than in TM. In conclusion, the core miRNAome identified in the samples from CM and TM may play an important role in cell proliferation, bone homeostasis, and neuronal network formation in newborn goat kids. The lack of differential miRNA expression between the CM and TM samples may be due to a relatively short sampling interval in which diet composition, intake and health status of ewes, and environment were relatively stable.

Characterization of the circulating transcriptome expression profile and identification of novel miRNA biomarkers in hypertrophic cardiomyopathy

BackgroundHypertrophic cardiomyopathy (HCM), one of the most common genetic cardiovascular diseases, but cannot be explained by single genetic factors. Circulating microRNAs (miRNAs) are stable and highly conserved. Inflammation and immune response participate in HCM pathophysiology, but whether the miRNA profile changes correspondingly in human peripheral blood mononuclear cells (PBMCs) with HCM is unclear. Herein, we aimed to investigate the circulating non-coding RNA (ncRNA) expression profile in PBMCs and identify potential miRNAs for HCM biomarkers.MethodsA Custom CeRNA Human Gene Expression Microarray was used to identify differentially expressed (DE) mRNAs, miRNAs, and ncRNAs (including circRNA and lncRNA) in HCM PBMCs. Weighted correlation network analysis (WGCNA) was used to identify HCM-related miRNA and mRNA modules. The mRNAs and miRNAs from the key modules were used to construct a co-expression network. Three separate machine learning algorithms (random forest, support vector machine, and logistic regression) were applied to identify potential biomarkers based on miRNAs from the HCM co-expression network. Gene Expression Omnibus (GEO) database (GSE188324) and experimental samples were used for further verification. Gene set enrichment analysis (GSEA) and competing endogenous RNA (ceRNA) network was used to determine the potential functions of the selected miRNAs in HCM.ResultsWe identified 1194 DE-mRNAs, 232 DE-miRNAs and 7696 DE-ncRNAs in HCM samples compared with normal controls from the microarray data sets. WGCNA identified key miRNA modules and mRNA modules evidently associated with HCM. We constructed a miRNA‒mRNA co-expression network based on these modules. A total of three hub miRNAs (miR-924, miR-98 and miR-1) were identified by random forest, and the areas under the receiver operator characteristic curves of miR-924, miR-98 and miR-1 were 0.829, 0.866, and 0.866, respectively.ConclusionsWe elucidated the transcriptome expression profile in PBMCs and identified three hub miRNAs (miR-924, miR-98 and miR-1) as potential biomarkers for HCM detection.

Exosome miR-552 Promotes Laryngocarcinoma Cells Malignant Progression via the PTEN/TOB1 Axis

Introduction: Tumor exosome-derived miRNAs play important roles in the human laryngocarcinoma. However, it is still unknown if exosome miR-552 is involved in the laryngocarcinoma. The aim of the current study was to explore exosome miR-552’s role in laryngocarcinoma and its underlying mechanisms. Methods: Hep-2 exosome was characterized by transmission electron microscopy and nanoparticle tracking technology. CCK-8 was used to determine cell viability, and a xenograft animal model was used to determine the tumorigenicity. qPCR and Western blotting were used to measure the changes in target biomarkers. Luciferase reporter assay was used to evaluate the interactions between miR-552 and PTEN. miRNA sequencing was used to check the changes in miRNA profiles. Results: miR-552 was upregulated in the laryngocarcinoma patients and was positively correlated to the cell proliferation and tumor growth. PTEN was identified as a direct target of miR-552. Hep-2 exosome is featured by high expression of miR-552 and treatment of Hep-2 exosome enhanced cell proliferation and tumorigenicity. The underlying mechanisms revealed that treatment of exosomes enhanced the malignant transformation of recipient cells in part by regulating epithelial-mesenchymal transition. Conclusion: Exosome miR-552 promotes laryngocarcinoma cells’ malignant progression in part by the regulation of the PTEN/TOB1 axis.

The role of miRNAs in the development of hepatobiliary diseases

The role of miRNA in the development of hepatobiliary diseases is considered in the scientific review. To write the article, information was searched using Scopus, Web of Science, MedLine, PubMed, Google Scholar, EMBASE, Global Health, The Cochrane Library databases. The authors provided information on the generation of a certain spectrum of miRNA by different types of cells of the hepatobiliary system. The authors state that miR-122-5p, miR-101, miR-192, miR-193, miR-194, miR-802 are hepatocyte-associated microRNAs. It is known that numerous microRNAs are involved in the regulation of the inflammatory process: miR-21, miR-29a, miR-96, miR-122, miR-125b, miR-132, miR-155, miR-146a, miR-150, miR-155, miR-181 and let-7. Among the many microRNAs involved in the regulation of inflammation in the liver, miR-122 and miR-155 are the most studied. It is stated that microRNAs such as let-7d, miR-15b, miR-19b, miR-21, miR-24, miR-29c, miR-122, miR-145, miR-155, miR-199, miR-200c, miR-378a are the most important modulators for the activity of fibrosis development in various tissues of the body, including liver tissue. MicroRNAs involved in the development of TGF-β1-induced liver fibrosis are presented. Scientists believe that since microRNAs regulate the cell cycle, proliferation and apoptosis of cells, they are factors that significantly affect the development of hepatobiliary tumors. It is emphasized that changes in miRNA profile in liver tissue or blood serum have their diagnostic value. Modulations of microRNAs are shown, which are associated with specific diseases of the liver and biliary tract with a high degree of probability. Thus, various miRNAs play an important role in the development of hepatobiliary diseases. A certain spectrum of miRNAs is generated by different cell types and plays an important role in the maturation and functioning of the hepatobiliary system. MicroRNAs indirectly participate in the synthesis of proteins, differentiation of cells, tissues and play an important pathogenetic role in the development of hepatobiliary diseases. The most important feature of microRNAs is their high resistance to the action of enzymatic substances, which allows them to be used as diagnostic or prognostic biomarkers.

Reprogramming of Fundamental miRNA and Gene Expression during the Barley-Piriformospora indica Interaction.

The interactions between plants and microorganisms, which are widely present in the microbial-dominated rhizosphere, have been studied. This association is highly beneficial to the organisms involved, as plants benefit soil microorganisms by providing them with metabolites, while microorganisms promote plant growth and development by promoting nutrient uptake and/or protecting the plant from biotic and abiotic stresses. Piriformospora indica, an endophytic fungus of Sebacinales, colonizes the roots of a wide range of host plants and establishes various benefits for the plants. In this work, an interaction between barley and the P. indica was established to elucidate microRNA (miRNA)-based regulatory changes in miRNA profiles and gene expression that occurred during the symbiosis. Growth promotion and vigorous root development were confirmed in barley colonized by P. indica. The genome-wide expression profile analysis of miRNAs in barley root showed that 7,798,928, 6,418,039 and 7,136,192 clean reads were obtained from the libraries of mock, 3 dai and 7 dai roots, respectively. Sequencing of the barley genome yielded in 81 novel miRNA and 450 differently expressed genes (DEGs). Additionally, 11, 24, 6 differentially expressed microRNAs (DEMs) in barley were found in the three comparison groups, including 3 dai vs. mock, 7 dai vs. mock and 7 dai vs. 3 dai, respectively. The predicted target genes of these miRNAs are mainly involved in transcription, cell division, auxin signal perception and transduction, photosynthesis and hormone stimulus. Transcriptome analysis of P. indica identified 667 and 594 differentially expressed genes (DEG) at 3 dai and 7 dai. Annotation and GO (Gene Ontology) analysis indicated that the DEGs with the greatest changes were concentrated in oxidoreductase activity, ion transmembrane transporter activity. It implies that reprogramming of fundamental miRNA and gene expression occurs both in barley and P. indica. Analysis of global changes in miRNA profiles of barley colonized with P. indica revealed that several putative endogenous barley miRNAs expressed upon colonization belonging to known micro RNA families involved in growth and developmental regulation.

Age-associated changes in miRNA profile of bovine follicular fluid.

This study shows that ageing affects miRNA profiles in follicular fluid, and an miRNA that is highly abundant in the follicular fluid of young cows supports the growth of oocytes derived from early antral follicles. We examined age-associated changes in miRNA profiles in the follicular fluid (FF) of cows. The role of miR-19b, which is abundant in the FF of young cows, in in vitro growth of early antral follicles (EAFs)-derived oocytes was assessed. FF was collected from the antral follicles of young (20-40 months) and aged (>120 months) cows. The miRNA profiles were similar between the FF of both age groups, whereas the abundance of some miRNAs differed between these samples. The miRNA profiles in granulosa cells (GCs) and the spent culture medium of oocyte-GC complexes (OGCs) derived from EAFs were distinct. Some miRNA groups overlapped among the GCs, culture media, and FFs. miR-19b was highly abundant in the FF of young cows, GCs, and culture medium. The supplementation of OGC culture medium with miR-19b increased the diameter, acetylation levels, and fertilisation ability of the oocytes. To assess whether miR-19b was functional in the GCs, a dual-luciferase assay, suppression of target protein, and RNA-sequencing of the GCs followed by functional annotation of the differentially expressed genes (DEGs) were conducted. Functional annotation of the DEGs suggested that miR-19b influences genes associated with FoxO signalling, endocytosis, and NR3C1 in GCs. These results suggest that in FFs, ageing affects the abundance of miRNAs that have important roles in oocyte development.

Circulating and skeletal muscle microRNA profiles are more sensitive to sustained aerobic exercise than energy balance in males.

MicroRNAs (miRNAs) regulate molecular processes governing muscle metabolism. Physical activity and energy balance influence both muscle anabolism and substrate metabolism, but whether circulating and skeletal muscle miRNAs mediate those effects remains unknown. This study assessed the impact of sustained physical activity with participants in energy balance (BAL) or deficit (DEF) on circulating and skeletal muscle miRNAs. Using a randomized cross-over design, 10 recreational active healthy males (mean±SD, 22±5 years, 87±11kg) completed 72h of high aerobic exercise-induced energy expenditures in BAL (689±852kcal/day) or DEF (-2047±920kcal/day). Blood and muscle samples were collected under rested/fasted conditions before (PRE) and immediately after 120min load carriage exercise bout at the end (POST) of the 72h. Trials were separated by 7days. Circulating and skeletal muscle miRNAs were measured using microarray RT-qPCR. Independent of energy status, 36 circulating miRNAs decreased (P<0.05), while 10 miRNAs increased and three miRNAs decreased in skeletal muscle (P<0.05) at POST compared to PRE. Of these, miR-122-5p, miR-221-3p, miR-222-3p and miR-24-3p decreased in circulation and increased in skeletal muscle. Two circulating (miR-145-5p and miR-193a-5p) and four skeletal muscle (miR-21-5p, miR-372-3p, miR-34a-5p and miR-9-5p) miRNAs had time-by-treatment effects (P<0.05). These data suggest that changes in miRNA profiles are more sensitive to increased physical activity compared to energy status, and that changes in circulating miRNAs in response to high levels of daily aerobic exercise are not reflective of changes in skeletal muscle miRNAs. KEY POINTS: Circulating and skeletal muscle miRNA profiles are more sensitive to high levels of aerobic exercise-induced energy expenditure compared to energy status. Changes in circulating miRNA in response to high levels of daily sustained aerobic exercise are not reflective of changes in skeletal muscle miRNA.

MiRNAs as Predictive Factors in Early Diagnosis of Gestational Diabetes Mellitus.

IntroductionCirculating miRNAs are important mediators in epigenetic changes. These non-coding molecules regulate post-transcriptional gene expression by binding to mRNA. As a result, they influence the development of many diseases, such as gestational diabetes mellitus (GDM). Therefore, this study investigates the changes in the miRNA profile in GDM patients before hyperglycemia appears.Materials and MethodsThe study group consisted of 24 patients with GDM, and the control group was 24 normoglycemic pregnant women who were matched for body mass index (BMI), age, and gestational age. GDM was diagnosed with an oral glucose tolerance test between the 24th and 26th weeks of pregnancy. The study had a prospective design, and serum for analysis was obtained in the first trimester of pregnancy. Circulating miRNAs were measured using the NanoString quantitative assay platform. Validation with real time-polymerase chain reaction (RT-PCR) was performed on the same group of patients. Mann-Whitney U-test and Spearman correlation were done to assess the significance of the results.ResultsAmong the 800 miRNAs, 221 miRNAs were not detected, and 439 were close to background noise. The remaining miRNAs were carefully investigated for their average counts, fold changes, p-values, and false discovery rate (FDR) scores. We selected four miRNAs for further validation: miR-16-5p, miR-142-3p, miR-144-3p, and miR-320e, which showed the most prominent changes between the studied groups. The validation showed up-regulation of miR-16-5p (p<0.0001), miR-142-3p (p=0.001), and miR-144-3p (p=0.003).ConclusionWe present changes in miRNA profile in the serum of GDM women, which may indicate significance in the pathophysiology of GDM. These findings emphasize the role of miRNAs as a predictive factor that could potentially be useful in early diagnosis.

Integrated analysis of blood mRNAs and microRNAs reveals immune changes with age in the Yangtze finless porpoise (Neophocaena asiaeorientalis).

Populations of Yangtze finless porpoises (YFPs) have rapidly declined in recent decades, raising the specter of extinction. In order to protect YFPs, a greater understanding of their biology is needed, including studying how their immune functioning changes with age. Here, we systematically studied the hematologic and biochemical parameters, as well as mRNAs and miRNAs profiles of old, adult, and young YFPs. The lymphocyte (LYMPH), neutrophils (NEUT) and eosinophils (EOS) counts in old YFPs were lower than those in young or adult YFPs. When comparing old to adult YFPs, the latter showed higher expression of genes associated with the innate and adaptive immune systems, including complement components, major histocompatibility complex, interleukins, TNF receptors, and chemokines/cytokines. When comparing old to young YFPs, the most striking difference was in higher toll-like receptor signaling in the latter. When comparing adult to young YFPs, the former exhibited higher expression of genes related to adaptive immunity and the FoxO signaling pathway, but lower expression of genes associated with the PI3K-Akt signaling pathway. Negative miRNA-mRNA interactions were predicted in comparisons of the old and adult (326), old and young (316), adult and young (211) groups. Overall, these results delineate a progression from early innate immune function dominance to adaptive immune function enhancement (young to adult) and deterioration (adult to old), and the changes in miRNAs profile correlate with the effects of age on immune functions. This study is the first to observe the changes of immune function of Yangtze finless porpoise with age using transcriptome method, and the study's findings are of great significance for protecting this endangered species.

Combined drought and heat stresses trigger different sets of miRNAs in contrasting potato cultivars.

MicroRNAs are small, non-coding RNAs that are responsible for regulation of gene expression during plant growth and development. Although there are many studies on miRNAs in other plants, little work has been done to understand the role of miRNAs in abiotic stress tolerance in potatoes. This study investigates changes in miRNA profiles of two different potato cultivars (tolerant, Unica and susceptible, Russet Burbank) in response to heat, drought and their combination. Transcriptomic studies revealed that miRNA profiles depend on the susceptibility and tolerance of the cultivar and also the stress conditions. Large number of miRNAs were expressed in Unica, whereas Russet Burbank indicated lesser number of changes in miRNA expression. Physiological and transcriptional results clearly supported that Unica cultivar is tolerant to combined drought and heat stress compared to Russet Burbank. Moreover, psRNATarget analysis predicted that major miRNAs identified were targeting genes playing important roles in response to drought and heat stress and their important roles in genetic and post-transcriptional regulation, root development, auxin responses and embryogenesis were also observed. This study focused on eight miRNAs (Novel_8, Novel_9, Novel_105, miR156d-3p, miR160a-5p, miR162a-3p, miR172b-3p and miR398a-5p) and their putative targets where results indicate that they may play a vital role at different post-transcriptional levels against drought and heat stresses. We suggest that miRNA overexpression in plants can lead to increased tolerance against abiotic stresses; furthermore, there should be more emphasis on the studies to investigate the role of miRNAs in combined abiotic stress in plants.

Arginine Alters miRNA Expression Involved in Development and Proliferation of Rat Mammary Tissue.

Simple SummaryMiRNAs are small noncoding RNAs that regulate a variety of developmental and physiological processes, with many having well-defined developmental and cell-type specific expression patterns. Aspects of the cell cycle such as cell differentiation, proliferation and apoptosis can be regulated by miRNA, underscoring an unexplored link between arginine supply and mammary tissue function during lactation. The specific objective of the present study was to determine miRNA profiles in mammary tissue at the end of lactation in response to enhanced dietary supply of arginine. Our results indicate that arginine may potentially be involved in the development of rat mammary glands through miRNA.This study was designed to determine the effects of dietary arginine on development and proliferation in rat mammary tissue through changes in miRNA profiles. Twelve pregnant Wistar rats were allocated randomly to two groups. A basal diet containing arginine or the control diet containing glutamate on an equal nitrogen basis as the arginine supplemented diet were used. The experiment included a pre-experimental period of four days before parturition and an experimental period of 17 days after parturition. Mammary tissue was collected for histology, RNA extraction and high-throughput sequencing analysis. The greater mammary acinar area indicated that arginine supplementation enhanced mammary tissue development (p < 0.01). MicroRNA profiling indicated that seven miRNA (miR-206-3p, miR-133a-5p, miR-133b-3p, miR-1-3p, miR-133a-3p, miR-1b and miR-486) were differentially expressed in response to Arginine when compared with the glutamate-based control group. In silico gene ontology enrichment and KEGG pathway analysis revealed between 240 and 535 putative target genes among the miRNA. Further verification by qPCR revealed concordance with the differential expression from the sequencing results: 17 of 28 target genes were differentially expressed (15 were highly expressed in arginine and 2 in control) and 11 target genes did not have significant difference in expression. In conclusion, our study suggests that arginine may potentially regulate the development of rat mammary glands through regulating miRNAs.

Bone marrow stromal cells derived exosomal miR-10a and miR-16 may be involved in progression of patients with multiple myeloma by regulating EPHA8 or IGF1R/CCND1.

Interaction with bone marrow stromal cells (BMSCs) has been suggested as an important mechanism for the progression of multiple myeloma (MM) cells, while exosomes are crucial mediators for cell-to-cell communication. The study was to investigate the miRNA profile changes in exosomes released by BMSCs of MM patients and explore their possible function roles.The microarray datasets of exosomal miRNAs in BMSCs were downloaded from the Gene Expression Omnibus database (GSE110271: 6 MM patients, 2 healthy donors; GSE78865: 4 donors and 2 MM patients; GSE39571: 7 MM patients and 4 controls). The differentially expressed miRNAs (DEMs) were identified using the LIMMA method. The target genes of DEMs were predicted by the miRwalk 2.0 database and the hub genes were screened by constructing the protein–protein interaction (PPI) network, module analysis and overlapping with the differentially expressed genes (DEGs) after overexpression or knockout of miRNAs.Three downregulated DEMs were found to distinguish MM from normal and MM-MGUS controls in the GSE39571 dataset; one downregulated and one upregulated DEMs (hsa-miR-10a) could differentiate MM from normal and MM-MGUS controls in the GSE110271-GSE78865 merged dataset. Furthermore, 11 downregulated (hsa-miR-16) and 1 upregulated DEMs were shared between GSE39571 and merged dataset when comparing MM with normal samples. The target genes were predicted for these 17 DEMs. PPI with module analysis showed IGF1R and CCND1 were hub genes and regulated by hsa-miR-16. Furthermore, EPHA8 was identified as a DEG that was downregulated in MM cells when the use of has-miR-10a mimics; while IGF1R, CCND1, CUL3, and ELAVL1 were also screened as DEGs that were upregulated in MM cells when silencing of hsa-miR-16.BMSCs-derived exosomal miR-10a and miR-16 may be involved in MM progression by regulating EPHA8 or IGF1R/CCND1/CUL3/ELAVL1, respectively. These exosomal miRNAs or genes may represent potential biomarkers for diagnosis of MM and prediction of progression and targets for developing therapeutic drugs.

Sustained Morphine Delivery Suppresses Bone Formation and Alters Metabolic and Circulating miRNA Profiles in Male C57BL/6J Mice.

Opioid use is detrimental to bone health, causing both indirect and direct effects on bone turnover. Although the mechanisms of these effects are not entirely clear, recent studies have linked chronic opioid use to alterations in circulating miRNAs. Here, we developed a model of opioid-induced bone loss to understand bone turnover and identify candidate miRNA-mediated regulatory mechanisms. We evaluated the effects of sustained morphine treatment on male and female C57BL/6J mice by treating with vehicle (0.9% saline) or morphine (17 mg/kg) using subcutaneous osmotic minipumps for 25 days. Morphine-treated mice had higher energy expenditure and respiratory quotient, indicating a shift toward carbohydrate metabolism. Micro-computed tomography (μCT) analysis indicated a sex difference in the bone outcome, where male mice treated with morphine had reduced trabecular bone volume fraction (Tb.BV/TV) (15%) and trabecular bone mineral density (BMD) (14%) in the distal femur compared with vehicle. Conversely, bone microarchitecture was not changed in females after morphine treatment. Histomorphometric analysis demonstrated that in males, morphine reduced bone formation rate compared with vehicle, but osteoclast parameters were not different. Furthermore, morphine reduced bone formation marker gene expression in the tibia of males (Bglap and Dmp1). Circulating miRNA profile changes were evident in males, with 14 differentially expressed miRNAs associated with morphine treatment compared with two differentially expressed miRNAs in females. In males, target analysis indicated hypoxia-inducible factor (HIF) signaling pathway was targeted by miR-223-3p and fatty acid metabolism by miR-484, -223-3p, and -328-3p. Consequently, expression of miR-223-3p targets, including Igf1r and Stat3, was lower in morphine-treated bone. In summary, we have established a model where morphine leads to a lower trabecular bone formation in males and identified potential mediating miRNAs. Understanding the sex-specific mechanisms of bone loss from opioids will be important for improving management of the adverse effects of opioids on the skeleton. © 2022 American Society for Bone and Mineral Research (ASBMR).

Analysis of altered miRNA profiling in the colon of a mouse model with β-lactoglobulin allergy

ObjectivesThe differences in the expression profiles of colonic miRNAs between β-lactoglobulin (β-Lg) allergic mice and normal mice were analyzed to investigate the important role of the miRNA regulation mechanism in the pathogenesis of cow’s milk allergy. MethodsThe present study performed Illumina sequencing to characterize the miRNA profile changes in mouse colon responding to β-Lg challenge. Target genes were predicted by TargetScan 50 and miRanda 3.3a algorithms and assessed by GO and KEGG analysis. The expression levels of selected miRNAs and cytokine production were verified by cell transfection and quantitative RT-PCR. ResultsA total of 15 miRNAs were diversely expressed between the colon of the normal and β-Lg-sensitized mice (P < 0.05, fold change of >1.50 or <0.67), including six up-regulated miRNAs and nine down-regulated miRNAs, among which seven miRNAs were validated using qRT-PCR. GO enrichment and KEGG pathway analyses further revealed that biological process, protein binding, cytoplasm and the pathways of cancer were significantly enriched, which were closely connected to the allergic inflammation development. Additionally, six key functional interaction pairs in β-Lg allergy were identified in miRNA prediction algorithms and verified using qRT-PCR. ConclusionsWe can conclude that our results suggested that the miRNAs regulation network participated in the pathogenesis of cow’s milk allergy.

Impact of Continuous Flow Left Ventricular Assist Device on Molecular Genetic Changes in Aortic Tissue

A pulsatility decrement caused by implantation of continuous flow left ventricular assist device (LVAD) has been corroborated as an additive factor further compromising aortic wall morphology and function. MicroRNAs (miRNAs) are important post-transcriptional regulators of gene expression which could reflect individual biologic adaptation to exposure to the low pulsatile environment. We hypothesized that pulsatility deficit caused by CF-LVAD implantation may influence miRNAs expression in aortic wall and that the miRNA profiles may differ between devices with different pulsatility profiles, such as speed modulated artificial pulse wave in HeartMate3. Paired aortic samples obtained at the time of LVAD implantation and at the time of Heart transplantation from CF-LVAD patients were examined for miRNA profiling in ten patients (3 females; mean age 55±13.8 years, ischemic cardiomyopathy 40%; HeartMateII(n=4), HeartMate3 (n=6). Human miRNome profile (miRCURY LNA miRNA PCR panels) of 752 miRNAs using qPCR on QuantStudio6 RT-PCR instrument was performed in all samples. The mean support duration was 383.5±222.4 days (range 162 to 887 days). 25 differently expressed multifunctional miRNAs that contribute to the regulation of multiple signaling pathways were identified. The most significant differences in miRNAs expression between implant/explant samples were found in miR-99b-5p; -144-3p; -23b-3p; -30e-3p and let-7d-5p (all p<0.002) which all play important role in differentiation of endothelial cells, vascular smooth muscle cells, or inflammation. No differences were observed between different types of CF-LVAD. The study provides additional evidence on a negative effect of suppressed physiologic pulsatility amplitude in CF-LVAD patients. Negative changes predominantly in miRNAs participating in vascular tissue engineering in aortic wall and pro-inflammatory changes of miRNA profiles during LVAD duration were observed. Our observations suggest that a programmed artificial pulsatility in HeartMate3 does not provide mitigation of continuous flow effect on aortic wall. Moreover, the most up-expressed miR-99b which target ENO2 gene, a marker of neuronal injury, was detected after explant of LVAD. Larger scale adverse events matched clinical trials are warranted.

Tspan8-Tumor Extracellular Vesicle-Induced Endothelial Cell and Fibroblast Remodeling Relies on the Target Cell-Selective Response.

Tumor cell-derived extracellular vesicles (TEX) expressing tetraspanin Tspan8-alpha4/beta1 support angiogenesis. Tspan8-alpha6/beta4 facilitates lung premetastatic niche establishment. TEX-promoted target reprogramming is still being disputed, we explored rat endothelial cell (EC) and lung fibroblast (Fb) mRNA and miRNA profile changes after coculture with TEX. TEX were derived from non-metastatic BSp73AS (AS) or metastatic BSp73ASML (ASML) rat tumor lines transfected with Tspan8 (AS-Tspan8) or Tspan8-shRNA (ASML-Tspan8kd). mRNA was analyzed by deep sequencing and miRNA by array analysis of EC and Fb before and after coculture with TEX. EC and Fb responded more vigorously to AS-Tspan8- than AS-TEX. Though EC and Fb responses differed, both cell lines predominantly responded to membrane receptor activation with upregulation and activation of signaling molecules and transcription factors. Minor TEX-initiated changes in the miRNA profile relied, at least partly, on long noncoding RNA (lncRNA) that also affected chromosome organization and mRNA processing. These analyses uncovered three important points. TEX activate target cell autonomous programs. Responses are initiated by TEX targeting units and are target cell-specific. The strong TEX-promoted lncRNA impact reflects lncRNA shuttling and location-dependent distinct activities. These informations urge for an in depth exploration on the mode of TEX-initiated target cell-specific remodeling including, as a major factor, lncRNA.

MiRNA profile in normal and aniridia conjunctival cells versus severity of aniridia‐associated keratopathy

AbstractPatients with congenital aniridia develop aniridia‐associated keratopathy (AAK), most of them exhibiting a similar disease mechanism namely haploinsufficiency of the PAX6 gene. The clinical onset and progression of AAK is quite diverse across patients. MiRNAs may act as regulatory elements affecting AAK progression. In mouse animal models the role of miRNAs controlling corneal vascularization has come into research focus. Interestingly, some of the miRNAs involved in neovascularization of the cornea, such as miR‐204, are regulated by PAX6 expression. The aim of this study was to identify deregulated miRNAs in conjunctival cells of AKK patients compared to unaffected controls, also in context with PAX6 haploinsufficiency. We further attempted to correlate miRNA expression with clinical severity of AKK (Staging ranging from 0 intact limbus, no pannus, clear cornea to 4 – end‐stage, thick, white, opaque vascularized pannus). We analyzed the miRNA expression profile in conjunctival cells of 20 patients with AKK and 20 age and sex matched controls using microarrays. We identified 10 down‐ and 11 upregulated miRNAs, including downregulated miR‐204‐5p and upregulated mir‐375. The latter miRNA has been reported to bind in PAX6 3′ UTR. Two patients showed mir‐204‐5p expression levels similar to healthy controls. These patients had mutations in PAX6 flanking genes likely affecting PAX6 regulatory regions and 0‐1 severity of AAK phenotype. Our results show strong expression changes in miRNA profile of Aniridia patients.