Project description:In this study, the methods to isolate and identify extracellular vesicles (EVs) including exosomes, from the seminal plasma (SP) of 3 fertile (F) and subfertile (S) bucks have been developed. Additionally, we investigated whether specific miRNA abundance differences between F and SF bucks could serve as fertility biomarkers. Ultracentrifugation and size exclusion chromatography analysis have made it possible to isolate different SP-EVs concentrations (8.53x10^11 ± 1.04x10^11 and 1.84x10^12 ± 1.75x10^11 particles/ml of SP; p=0,008), with a similar average size (143.9 ± 11.9 and 115.5 ± 2.4 nm; p=0.7422) in F and S males, respectively. Particle size was not significantly correlated with any kinetic parameter. Also, EVs have been identified by electron microscopy and their marker proteins by Western blot. The concentration of SP-EVs was positively correlated with the percentage of abnormal forms (r=0.94; p<0.05) and with the percentage of immotile spermatozoa (r=0.88; p<0.05). A total of 18 miRNAs were differentialy expressed (FC>2, FDR<0.05) in F vs. S groups. SP from F and S males contains EVs with different miRNA cargo that could be used as biomarkers for male fertility.
Project description:We studied extracellular vesicles (EVs) from synovial fibroblast (SF). EVs were isolated from the secretome of non-senescent and irradiation-induced senescent SFs using size exclusion chromatography. EV RNA was extracted and subject to small RNA sequencing on an Illumina NovaSeq SP using 100bp, single end reads. After mapping against GRCh38.p12 and miRBase v22.1 differential expression analysis was undertaken with edgeRv3.28 using a quasi-likelihood negative binomial generalized log-linear model. We identified a panel of differentially expressed small non-coding RNAs.
Project description:The goal of this study was to identify chromatin regulatory sites by FAIRE-seq under conditions of basal and increased dosage of transcription factor SF-1 in the H295R human adrenocortical tumor cell line. 4 samples: input DNA in basal SF-1 expression conditions - FAIRE-seq in basal SF-1 expression conditions - input DNA in SF-1 overexpression conditions - FAIRE-seq in SF-1 overexpression conditions
Project description:To explore the cross feeding interactions between the acetogenic human colonic bacterium Blautia hydrogenotrophica and the nutritional specialist amyloltic bacterium Ruminococcus bromii in a continuous culture system utilising transcriptome analysis.
Project description:The goal of this study was to identify genomic binding sites of the NRSF/REST transcription factor under conditions of basal and increased SF-1 dosage in the H295R human adrenocortical tumor cell line. 4 samples: input DNA (2 replicates) - NRSF/REST ChIP basal SF-1 dosage - NRSF/REST ChIP increased SF-1 dosage
Project description:Background: PLK1 is overexpressed in acute myeloid leukemia (AML). A Phase 1b trial of the PLK1 inhibitor onvansertib (ONV) combined with decitabine (DAC) demonstrated initial safety and efficacy in patients with relapsed/refractory (R/R) AML. The current study aimed to identify molecular predictors of response to ONV + DAC in R/R AML patients. Patients and methods: A total of 44 R/R AML patients were treated with ONV+DAC and considered evaluable for efficacy. Bone marrow (BM) samples were collected at baseline for genomic and transcriptomic analysis (n=32). A 10 gene expression-signature, predictive of response to ONV + DAC, was derived from the leading-edge genes of gene set enrichment analyses (GSEA). The gene signature was evaluated in independent datasets and used to identify associated mutated genes. Results: 20% of the patients achieved complete remission, with or without hematologic count recovery (CR/CRi) and 32% exhibited a ≥50% reduction in bone marrow blasts. Patients who responded to treatment had elevated mitochondrial function and OXPHOS. The gene signature was not associated with response to DAC alone in an independent dataset. By applying the signature to the BeatAML cohort (n=399), we identified a positive association between predicted ONV + DAC response and mutations in splicing factors (SF). In the Phase 1b/2 trial, patients with SF mutations (SRSF2, SF3B1) had a higher CR/CRi rate (50%) compared to those without SF mutations (9%). Conclusion: PLK1 inhibition with ONV in combination with DAC could be a potential therapy in R/R AML patients, particularly those with high OXPHOS gene expression and SF mutations.
