Idiopathic central precocious puberty (CPP) is increasingly observed in girls. Premature thelarche (PT) and exaggerated thelarche (ET) are early pubertal variants that can be challenging to distinguish from CPP in clinical practice. Exosomal microRNAs are stable biomarkers capable of crossing the blood-brain barrier. Although miR-30b-5p has been reported to increase in pubertal boys and girls, human studies investigating microRNAs in CPP and puberty remain limited. To investigate exosomal microRNA expression profiles and associated pathways in early pubertal development, we conducted a cross-sectional study of 28 girls aged 6-8 years. Serum exosomal microRNA expression was analyzed using next-generation sequencing. Differentially expressed microRNAs (DEmiRNAs) between groups were identified, followed by pathway enrichment analysis. Distinct exosomal miRNA expression patterns were observed among the CPP, ET, and control groups, with 307 DEmiRNAs identified. The CPP, PT, and ET groups exhibited distinct miRNA expression profiles compared with the control group. miR-30b-5p was upregulated in the CPP, ET, and PT groups compared with the control group. Pathway enrichment analysis revealed the involvement of various signaling pathways including AGE-RAGE, MAPK, and mTOR signaling pathways. Serum exosomal microRNAs may serve as biomarkers for early puberty and provide insight into metabolic influences on pubertal development.

Extracellular vesicles (EVs) facilitate intercellular communication through the transfer of bioactive molecules that modulate tumor behavior. In this study, we demonstrate that oncogenic KRAS- and BRAF-mutations alter EV composition, modify surface properties, and impact intracellular trafficking. Through comparative proteomics, uptake assays, and nano-flow cytometry, we found that mutant EVs display increased levels of CD44, LSR, ITGB4, and CSPG4, leading to enhanced uptake and preferential trafficking to the endoplasmic reticulum and nucleus, while avoiding lysosomal degradation. Disruption of CD44 and CSPG4-mediated interactions with hyaluronic acid or chondroitin sulfate, respectively, reduced EV internalization and lysosomal trafficking, verifying their roles in mutation-dependent uptake. Notably, doxorubicin-loaded mutant EVs showed improved nuclear delivery and increased cytotoxicity compared to wild-type EVs, indicating that mutation-specific EVs function as efficient drug delivery vehicles. Collectively, these results reveal a direct association between oncogenic signaling and EV diversity, highlighting the potential of mutation-adapted EVs for precise delivery of therapeutics to specific subcellular compartments.

Purpose: Prostate cancer is one of the most common malignancies in men, yet current prognostic methods remain suboptimal. Emerging evidence indicates that microRNAs (miRNAs) play critical roles in prostate cancer progression. This study aimed to identify miRNAs associated with adverse clinical outcomes by comparing miRNA expression profiles between prostate tumors with unfavorable versus favorable prognostic features. Materials and Methods: High-throughput next-generation sequencing (NGS) was used to analyze miRNA expression in formalin-fixed, paraffin-embedded prostate cancer tissue samples. Patients were classified into favorable or unfavorable prognosis groups based on risk stratification scores, Gleason grade group, and biochemical recurrence. Differentially expressed miRNAs were identified using a fold-change threshold ≥2 and a false discovery rate (FDR) <0.05. Predicted target genes and pathway analyses were conducted to generate candidate regulatory hypotheses rather than confirm mechanistic relationships. Results: Several miRNAs were differentially expressed according to prognostic category. miR-206 was significantly downregulated in high-risk tumors compared with low-risk tumors. High-Gleason-grade tumors showed reduced expression of miR-7704 and miR-4454, while miR-25-3p and let-7f-5p were upregulated. In patients with early biochemical recurrence, miR-7704 and miR-10400-5p were downregulated relative to those with prolonged recurrence-free survival. Target prediction analysis identified CPEB3, HAND1, PTAR1, and SPRYD4 as shared candidate targets, with CPEB3 emerging as a prioritized candidate supported by consistency in external datasets rather than a confirmed molecular target. Conclusions: Distinct miRNA expression patterns correlate with prostate cancer aggressiveness and clinical outcomes. miR-206, miR-7704, miR-4454, miR-25-3p, and let-7f-5p represent candidate prognostic biomarkers. Their shared target CPEB3 should be interpreted as a prioritized candidate for future investigation. Given the very small sample size and the lack of qRT-PCR and functional validation, these findings should be considered preliminary and hypothesis-generating, requiring validation in larger independent cohorts and experimental studies.

Background/Objectives: Prostate cancer (PCa) remains a leading cause of cancer-related mortality in men. While localized PCa carries a favorable prognosis, progression to locally advanced disease substantially worsens outcomes. There is an unmet need for molecular markers that reliably distinguish indolent from aggressive tumors. MicroRNAs (miRNAs), small non-coding RNAs that post-transcriptionally regulate gene expression, are implicated in cancer development and progression, and their expression profiles may serve as putative biomarkers of disease stage. Methods: We performed small RNA sequencing on formalin-fixed paraffin-embedded (FFPE) prostate tumor tissues from nine patients (five localized, four locally advanced-stage). Differentially expressed miRNAs were identified using a bioinformatics pipeline (fold-change ≥ 2, p < 0.05). Target gene prediction, KEGG pathway enrichment, and miRNA–mRNA interaction network analyses were conducted, followed by in silico validation using two independent public mRNA datasets (GSE46602 and GSE21034, a subseries of GSE21032). Results: Seven unique mature miRNAs were differentially expressed between localized and locally advanced PCa. Five miRNAs (miR-145-5p, miR-205-5p, miR-206, miR-24-1-5p, and hsa-miR-3648) were downregulated in locally advanced tumors, while hsa-miR-625-3p and miR-324-5p were upregulated. Network analysis identified CLTC and CDK6 as putative downstream targets co-targeted by multiple downregulated miRNAs. Independent validation using GSE46602 and GSE21034 consistently demonstrated significantly higher expression of CLTC and CDK6 in locally advanced prostate cancer following Benjamini–Hochberg correction. Conclusions: This exploratory pilot analysis identified several miRNAs that differ between localized (pT2) and locally advanced (pT3–T4/N+) prostate cancer. Independent validation across two publicly available mRNA cohorts supports CLTC and CDK6 as candidate stage-associated genes. However, these findings remain hypothesis-generating and require validation in larger prospective cohorts together with functional confirmation before biomarker or therapeutic claims can be made.

Background/Objectives: This NGS-based study sought to identify novel molecular markers for prostate cancer by comparing miRNA expression in cancer and benign prostatic hyperplasia (BPH) tissues. Methods: Using high-throughput sequencing and stringent statistical criteria, the study identified eleven significantly dysregulated miRNAs (five downregulated, six upregulated) that differentiate the two conditions. Enrichment analyses linked these miRNAs to several key cancer-associated pathways, including PI3K-Akt and ErbB signaling. Results: Crucially, the protein vesicle-associated membrane protein-associated protein B (VAPB) was pinpointed as a central hub, regulated by three downregulated miRNAs (miR-143-3p, miR-221-3p, and miR-222-3p). Since VAPB has not been widely studied in prostate cancer, it represents a promising, novel candidate for both diagnosis and therapeutic targeting. Conclusions: Our NGS-based analysis revealed a distinct miRNA expression signature that differentiates prostate cancer from BPH. The downregulation of several tumor-suppressive miRNAs (with concomitant upregulation of oncogenic miRNAs) in prostate cancer may contribute to malignancy-including the de-repression of novel targets like VAPB, which we identify as a promising new biomarker and therapeutic target.

Extracellular vesicles (EVs) are implicated in the pathophysiology of obesity and diabetes. This study aimed to analyze the impact of diabetes on EV protein dynamics in patients with obesity before and after bariatric surgery. This prospective study assessed the serum EV proteins in 30 patients with obesity, with and without diabetes, and 37 healthy controls using liquid chromatography-tandem mass spectrometry. EV proteins were also analyzed 6 months after bariatric surgery in patients with obesity. We found 19 differentially expressed proteins (DEPs) in EVs between healthy controls and patients with obesity. In addition, 20 DEPs between patients with obesity who had and did not have diabetes were identified. After bariatric surgery, 14 DEPs mainly involved in the immune system were identified in patients with obesity and diabetes; adiponectin (ADIPOQ), mannose binding lectin 2 (MBL2), and hornerin (HRNR) were associated with glycemic control, body mass index, and weight loss, respectively. In patients with obesity without diabetes, 13 DEPs after bariatric surgery were predominantly involved in reactive oxygen species metabolism. Diabetes affects the differences in EV protein profiles in patients with obesity before and after bariatric surgery. These DEPs may serve as potential biomarkers and therapeutic targets for obesity-related diabetes.

Most cancer cells adopt a less efficient metabolic process of aerobic glycolysis with high level of glucose uptake followed by lactic acid production, known as the Warburg effect. This phenotypic transition enables cancer cells to achieve increased cellular survival and proliferation in a harsh low-oxygen tumor microenvironment. Also, the resulting acidic microenvironment causes inactivation of the immune system such as T-cell impairment that favors escape by immune surveillance. While lots of studies have revealed that tumor-derived EVs can deliver parental materials to adjacent cells and contribute to oncogenic reprogramming, their functionality in energy metabolism is not well addressed. In this study, we established prostate cancer cells PC-3AcT resistant to cellular death in an acidic culture medium driven by lactic acid. Quantitative proteomics between EVs derived from PC-3 and PC-3AcT cells identified 935 confident EV proteins. According to cellular adaptation to lactic acidosis, we revealed 159 regulated EV proteins related to energy metabolism, cellular shape, and extracellular matrix. These EVs contained a high abundance of glycolytic enzymes. In particular, PC-3AcT EVs were enriched with apolipoproteins including apolipoprotein B-100 (APOB). APOB on PC-3AcT EVs could facilitate their endocytic uptake depending on low density lipoprotein receptor of recipient PC-3 cells, encouraging increases of cellular proliferation and survival in acidic culture media via increased activity and expression of hexokinases and phosphofructokinase. The activation of recipient PC-3 cells can increase glucose consumption and ATP generation, representing an acquired metabolic reprogramming into the Warburg phenotype. Our study first revealed that EVs derived from prostate cancer cells could contribute to energy metabolic reprogramming and that the acquired metabolic phenotypic transition of recipient cells could favor cellular survival in tumor microenvironment.

BackgroundMicroRNAs (miRNAs) have emerged as potential biomarkers for insulin resistance-related conditions such as obesity, metabolic syndrome, and metabolic dysfunction-associated steatotic liver disease (MASLD) in adults. However, data in pediatric populations remain limited. This study aimed to compare serum exosomal miRNA expression among children with both obesity and MASLD, children with obesity without MASLD, and healthy controls, and to evaluate associations with insulin resistance markers.MethodsThirty prepubertal children (5 males and 5 females per group) were enrolled: children with both obesity and MASLD (n=10), children with obesity without MASLD (n=10), and healthy controls (n=10). Serum exosomal miRNAs were analyzed using next-generation sequencing. Differential expression analyses were performed between groups, and correlations with clinical and metabolic parameters were assessed, including the triglyceride-glucose (TyG) index, TyG index adjusted for ALT (TyG-ALT), and the homeostatic model assessment for insulin resistance (HOMA-IR).ResultsAll participants were prepubertal, with no significant differences in mean age across groups. miR-34a-5p, miR-122-5p, miR-885-5p, and miR-885-3p were significantly upregulated in children with both obesity and MASLD compared to both other groups, and positively correlated with AST and uric acid levels. Additionally, miR-122-5p, miR-885-5p, and miR-885-3p were associated with ALT and TyG-ALT index. Conversely, miR-570-3p and miR-32-3p were significantly downregulated in children with both obesity and MASLD and negatively correlated with AST, ALT, and TyG-ALT levels.ConclusionsSix exosomal miRNAs (miR-34a-5p, miR-122-5p, miR-885-5p, miR-885-3p, miR-570-3p, and miR-32-3p) were differentially expressed in children with both obesity and MASLD and showed strong correlations with markers of liver function and insulin resistance. These findings suggest their potential as non-invasive biomarkers for early identification of MASLD and related metabolic disturbances in pediatric populations.

Background Type 2 diabetes is a complex metabolic disorder characterized by insulin resistance and progressive beta-cell dysfunction. Although sex differences in type 2 diabetes prevalence, progression, and complications have been reported, the molecular mechanisms underlying these differences remain largely unknown. We aimed to utilize single-cell RNA sequencing to identify a beta-cell cluster that is more prevalent in males than in females and exhibits distinct gene expression patterns, gene set enrichment profiles, and cell-cell communication compared to other clusters. Methods FASTQ files from four public datasets were preprocessed, aligned to the human genome (GRCh38), and integrated into a high-quality matrix to mitigate batch effects. We focused on beta-cells from type 2 diabetes patients, performed trajectory inference to identify clusters, and conducted differential gene expression and gene set enrichment analyses. These findings were validated using bulk RNA-seq datasets. Additionally, cell-cell communication analysis was performed to identify ligand-receptor interactions, followed by a sensitivity analysis to assess sex-specific differences. Results We identified a male-dominant beta-cell cluster (adjusted P value=4.2×10–6) that displayed unique gene expression patterns and downregulation of pathways associated with protein metabolism and insulin synthesis. Differentially expressed genes (e.g., interleukin 24 [IL24], regulator of G protein signaling like 1 [RGSL1]) were confirmed through bulk analysis. Moreover, the cluster demonstrated distinct communication patterns with other cell types, underscoring sex-specific differences. Conclusion We have identified a male-dominant beta-cell cluster characterized by distinct gene expression, signaling pathways, and cell interactions. These findings provide insights into the pathophysiology of type 2 diabetes and may inform the development of more effective, sex-specific therapeutic strategies in the future.

Early detection and surgical excision of tumors have helped improve the survival rate of patients with breast cancer. However, patients with metastatic cancer typically have a poor prognosis. In this study, we propose that ANKRD1 promotes metastasis of breast cancer. ANKRD1 was found to be highly expressed in the MDA-MB-231 and MDA-LM-2 highly metastatic breast cancer cell lines compared to the non-metastatic breast cancer cell lines (MCF-7, ZR-75-30, T47D) and normal breast cancer cells (MCF-10A). Furthermore, high-grade tumors showed increased levels of ANKRD1 compared to low-grade tumors. Both in vitro and in vivo functional studies demonstrated the essential role of ANKRD1 in cancer cell migration and invasion. The previous studies have suggested a significant role of NF-κB and MAGE-A6 in breast cancer metastasis, but the upstream regulators of this axis are not well characterized. Our study suggests that ANKRD1 promotes metastasis of breast cancer by activating NF-κB as well as MAGE-A6 signaling. Our findings show that ANKRD1 is a potential therapeutic target and a diagnostic marker for breast cancer metastasis.

The 5-year survival rate of kidney cancer drops dramatically from 93% to 15% when it is metastatic. Metastasis constitutes for 30% of kidney cancer cases, in which clear cell renal cell carcinoma (ccRCC) is the most prominent subtype. By sequencing mRNA of ccRCC patient samples, we found that apolipoprotein L1 (APOL1) was highly expressed in tumors compared to their adjacent normal tissues. This gene has been previously identified in a large body of kidney disease research and was reported as a potential prognosis marker in many types of cancers. However, the molecular function of APOL1 in ccRCC, especially in metastasis, remained unknown. In this study, we modulated the expression of APOL1 in various renal cancer cell lines and analyzed their proliferative, migratory, and invasive properties. Strikingly, APOL1 overexpression suppressed ccRCC metastasis both in vitro and in vivo. We then explored the mechanism by which APOL1 alleviated ccRCC malignant progression by investigating its downstream pathways. APOL1 overexpression diminished the activity of focal adhesive molecules, Akt signaling pathways, and EMT processes. Furthermore, in the upstream, we discovered that miR-30a-3p could inhibit APOL1 expression. In conclusion, our study revealed that APOL1 play a role as a tumor suppressor in ccRCC and inhibit metastasis, which may provide novel potential therapeutic approaches for ccRCC patients.

Kidney cancer is the 14th most common cancer globally. The 5-year relative survival rate of kidney cancer at a localized stage is 92.9% and it declines to 17.4% in metastatic stage. Currently, the most accurate method of its diagnosis is tissue biopsy. However, the invasive and costly nature of biopsies makes it undesirable in many patients. Therefore, novel biomarkers for diagnosis and prognosis should be explored. Urinary extracellular vesicles (uEVs) are small vesicles (50–200 nm) in urine carrying nucleic acids, proteins and lipids as their cargos. These uEVs’ cargos can provide non-invasive alternative to monitor kidney health. In this review, we have summarized recent studies investigating potential use of uEVs’ cargos as biomarkers in kidney cancer for diagnosis, prognosis and therapeutic intervention.

Breastfeeding not only reduces infection-related morbidity, but also increases growth of preterm infants. Advantages of breast milk (BM) for preterm infants are significant. They continue to be studied. However, because not all preterm infants can receive breastfeeding, bovine-based infant formula (IF) is used as an alternative, which may increase the risk of several preterm complications. Exosomes isolated from biofluids are emerging as biomarkers in research of various diseases. Here, we characterized miRNA contents of exosomes in urine and serum samples of preterm infants who were BM and IF fed and performed transcriptomic analysis of small RNA libraries. We identified significantly up-regulated 6 miRNAs and 10 miRNAs, respectively. Gene Ontology (GO) analysis revealed that target genes of these miRNAs might participate in neuronal development, immunity modulation, detoxification of reactive oxygen species, and transmembrane exchange. Our data suggest that exosome-based systemic screening for preterm infants with breastfeeding might be a screening tool for identifying target molecules involved in therapy for preterm infants in neonatal intensive care unit (NICU) and for future application as nutraceutical formulations or pharmaceuticals.

Supplementary Figure S1 - PDF file 216K, Conditioned media from metastasis-incompetent PC3 cancer cells generate GFP+ CD11b+ BM-derived niches in the lungs

Supplementary Figure S7 - PDF file 58K, The seeding of tumor cells in the lungs is similar in Tsp1-/- BMT mice and WT BMT mice

Supplementary Figure S2 - PDF file 44K, Tsp-1 is expressed in the lungs of mice treated with metastasis-incompetent PC3 CM

Supplementary Figure S9 - PDF file 221K, Metastatic lesions in WT BMT mice exhibit reduced proliferation

Supplementary methods - PDF file 98K, Supplementary methods

Supplementary Figure S10 - PDF file 175K, Prosaposin modulates lung Tsp-1

Supplementary Figure S5 - PDF file 91K, Characterization of Tsp-1-/- bone marrow after transplantation