Leukemia is a heterogeneous group of hematological malignancies. Despite the enormous progress that has been made in the field of hemato-oncology in recent years, there are still many problems related to, among others, disease recurrence and drug resistance, which is why the search for ideal biomarkers with high clinical utility continues. Research shows that exosomes play a critical role in the biology of leukemia and are associated with the drug resistance, metastasis, and immune status of leukemias. Exosomes with their cargo of non-coding RNAs act as a kind of intermediary in intercellular communication and, at the same time, have the ability to manipulate the cell microenvironment and influence the reaction, proliferative, angiogenic, and migratory properties of cells. Exosomal ncRNAs (in particular, circRNAs and microRNAs) appear to be promising cell-free biomarkers for diagnostic, prognostic, and treatment monitoring of leukemias. This review examines the expression of exosomal ncRNAs in leukemias and their potential regulatory role in leukemia therapy but also in conditions such as disease relapse, drug resistance, metastasis, and immune status. Given the key role of ncRNAs in regulating gene networks and intracellular pathways through their ability to interact with DNA, transcripts, and proteins and identifying their specific target genes, defining potential functions and therapeutic strategies will provide valuable information.

Umbilical cord mesenchymal stromal cells (UC-MSCs) are a key element of regenerative medicine due to their ability to secrete growth factors that stimulate proliferation and angiogenesis, and modulate the inflammatory response. Despite their widespread use, the influence of the perinatal microenvironment on their biological properties remains poorly understood. The aim of this study was to assess the influence of pH and blood gas parameters in umbilical cord blood on the global transcriptomic profile of UC-MSCs and to analyze the correlation between the metabolic status of the newborn and the expression of key trophic factors: EGF, FGF2, FGFR1, FGFR3, GDNF, HGF, IGF1, NES, NGF, and PGF. Methods: The study was conducted in two stages. In the first phase, transcriptomic screening was performed using Affymetrix HuGene 2.0 ST microarray on cells isolated from three environmental groups defined by cord blood pH: acidic (pH < 7.35), physiological (7.35–7.39), and alkaline (pH ≥ 7.4). In the second phase, the results were validated using qPCR on an expanded study group (N = 50). Gene expression levels (RQ) were related to blood gas parameters (pH, pCO₂, pO₂, cHCO₃) and the presence of clinical features of threatened neonatal asphyxia. Results: Microarray analysis revealed that environmental pH acts as a molecular phenotypic switch. Under low pH conditions (<7.35), a shift in cell profile from proliferative to structural–migratory was observed. Significant overexpression of genes responsible for extracellular matrix (ECM) organization and adhesion (e.g., COMP, DCN, LUM, FMOD) was observed, while pathways related to cell cycle and cell division (↓CDK1, AURKA, TOP2A) were downregulated. qPCR validation confirmed these observations, demonstrating a strong positive correlation between blood pH and the expression of regenerative mediators: FGFR1 (r = 0.28), EGF (r = 0.30), NGF (r = 0.39), and IGF1 (r = 0.30). A negative correlation was also found between carbon dioxide pressure (pCO₂) and the expression of NGF, FGFR1, and EGF. A significant clinical finding was that in newborns diagnosed with threatened asphyxia, EGF, FGFR1, and NGF gene expression was significantly reduced, indicating impaired trophic potential of the cells in response to metabolic stress. Conclusions: These results indicate that cord blood gas parameters are critical regulators of the genetic activity of UC-MSCs. Metabolic and respiratory acidosis not only inhibit the cells’ proliferative potential but also force them into a matrix remodeling mode, permanently modifying their transcriptomic profile. This suggests that the neonatal acid–base status may serve as an objective indicator of the “biological quality” of isolated stromal cells, which has significant implications for their future applications in cell therapies.

Background: Mesenchymal stem cells (MSCs) exhibit a high capacity for differentiation, possess anti-inflammatory and proangiogenic properties, and stimulate the growth and proliferation of neighboring cells. MSCs are a promising tool in regenerative medicine. However, the molecular mechanisms underlying the properties of these cells are not yet fully understood. Gene expression in MSCs influences their characteristics and differentiation potential. Therefore, it is essential to investigate factors affecting gene expression as well as those activating signaling pathways, which will enable more effective and individualized applications of MSCs. In this study, we aimed to identify signaling pathways involved in gene expression in umbilical cord-derived MSCs (UC-MSCs) that may be altered by maternal diabetes and hypothyroidism during pregnancy. Methods: The research material consisted of UC-MSCs. Samples obtained from nine participants were analyzed. UC-MSCs were isolated and cultured, and RNA was extracted. The isolated RNA was used for microarray-based gene expression analysis. Subsequently, pathway enrichment analysis was performed to identify the signaling pathways involved. Results: In the diabetes group, 340 genes (0.71%) were upregulated, while 268 genes (0.56%) were downregulated compared with UC-MSCs from the control group. In the diabetes group, the most compact module was composed of proteins associated with WNT/planar cell polarity (WNT/PCP) signaling. The second module included genes related to smooth muscle activity. In the hypothyroidism group, an association was identified between the extracellular matrix organization pathways (GO:0030198) and the extracellular structure organization (GO:0043062) pathways. Moreover, in this group, increased expression of MMP1, MMP10, and GREM1 was observed. Conclusions: In summary, our study demonstrated the impact of diabetes and hypothyroidism on gene expression in UC-MSCs. We also observed the activation of distinct signaling pathways depending on the presence of these conditions. However, this work represents a preliminary screening, and the results should be validated by PCR in a larger cohort.

Hypoxia-inducible factor 1 (HIF-1) may play a role in mammary gland development, milk production and secretion in mammals. Due to the limited number of scientific reports on the expression of HIF genes in colostrum cells, it was decided to examine the expression of HIF1A, HIF3A and EPAS1 in the these cells, collected from 35 patients who voluntarily agreed to provide their biological material for research, were informed about the purpose of the study and signed a consent to participate in it. The expression of HIF genes was assessed using qPCR. Additionally, the influence of clinical parameters (method of delivery, occurrence of stillbirths in previous pregnancies, BMI level before pregnancy and at the moment of delivery, presence of hypertension during pregnancy, presence of Escherichia coli in vaginal culture, iron supplement and heparin intake during pregnancy) on the gene expression was assessed, revealing statistically significant correlations. The expression of HIF1A was 3.5-fold higher in the case of patients with the presence of E. coli in vaginal culture (p = 0.041) and 2.5 times higher (p = 0.031) in samples from women who used heparin during pregnancy. Approximately 1.7-fold higher expression of the EPAS1 was observed in women who did not supplement iron during pregnancy (p = 0.046). To our knowledge, these are the first studies showing the relationship between HIF expression in cells from breast milk and the method of delivery and health condition of women giving birth. The assessment of HIF expression requires deeper examination in a larger study group, and the results of further studies will allow to determine whether HIF can become biomarkers in pregnancy pathology states.

Perinatal mesenchymal stem cells (MSCs) are characterized by high phenotypic plasticity, allowing them to dynamically adapt to microenvironmental stimuli. However, the extent to which maternal pathophysiology and clinical history in vivo permanently alter their biological properties remains poorly understood. The aim of this study is to determine the influence of the maternal in vivo environment—including vascular stress (gestational hypertension), metabolic stress (hypothyroidism), and obstetric history (miscarriages, stillbirths)—on the transcriptomic, proteomic, and epigenetic profiles of growth factors in umbilical cord blood stem cells (UC-MSCs) and human milk stem cells (MM-MSCs). UC-MSCs (n = 51) and MM-MSCs (n = 78) were isolated from eligible donors. Global transcriptomic pathways associated with maternal hypertension were examined in UC-MSCs using Affymetrix expression microarrays. Targeted polymerase chain reaction (qPCR) validation was performed in both cell sources for key growth factors and receptors and for microRNA-155 in MM-MSCs. Paracrine protein concentrations (HGF, IGF-1, VEGF) in whole milk were quantified using ELISA. Microarray screening demonstrated that maternal hypertension is associated with a transcriptomic shift in the UC-MSCs towards a profibrotic signature, marked by an increased expression of the canonical Wnt signaling pathway and structural components of the extracellular matrix, while simultaneously suppressing the expression of genes associated with cell migration networks. qPCR confirmed that maternal hypertension is associated with significantly reduced FGFR1 expression and a downward trend in EGF and HGF in UC-MSCs, while correlating with an asymmetric response in MM-MSCs. Maternal hypothyroidism was linked to a transcriptomic profile suggestive of a rescue response (a trend toward increased IGF1 and EGF expression) in UC-MSCs, in contrast to a potential paracrine translation deficit (trends toward reduced VEGF, HGF, and IGF-1 protein levels) in the milk matrix. Importantly, a history of miscarriage correlated with alterations in the cellular mechanism of MM-MSCs (a trend toward increased IGF1 and EGF mRNA expression), while previous stillbirths were associated with a systemic, threefold increase in free IGF-1 protein concentration in whole milk. This paracrine dynamic strongly correlated with miR-155 expression, suggesting a potential regulatory role in the lactation niche. Our findings suggest that the molecular and epigenetic makeup of perinatal stem cell niches is dynamically and persistently associated with the mother’s somatic health and reproductive history. This tissue-specific plasticity provides robust molecular support for the DOHaD paradigm, indicating that offspring stem cell profiles may directly reflect the maternal inheritance in vivo.

Tumor necrosis factor alpha-stimulated protein 6 (TSG-6) is a potent anti-inflammatory mediator secreted by mesenchymal stem cells (MSCs). In human milk (HM), these cells (human milk mesenchymal stem cells, HM-MSCs) contribute to postnatal programming of the neonatal gastrointestinal immune axis. This study assessed how maternal gestational hypertension and maternal age affect the cytoprotective TSG-6 pool and its associated immunomodulatory microRNA signature in early milk. Early colostrum, 2–4 days postpartum, from healthy mothers and hypertensive patients was analyzed. TSG-6 gene expression in the heterogeneous milk cellular fraction containing HM-MSCs and TSG-6 protein concentration in whole milk were determined using quantitative polymerase chain reaction (qPCR) and Enzyme-Linked Immunosorbent Assay (ELISA), respectively. Comprehensive multivariable General Linear Models (GLMs) were used to adjust for potential confounding variables, including pre-pregnancy Body Mass Index (BMI), gestational age (GA), and gestational diabetes. Additionally, TSG-6 levels were correlated with fetal cord blood gas parameters and previously profiled microRNAs in the cellular fraction. GLM analysis revealed that gestational hypertension remained significantly associated with reduced TSG-6 gene expression in the cellular fraction and total TSG-6 protein concentration in whole milk, regardless of pre-pregnancy BMI or metabolic comorbidities. In healthy pregnancies, exploratory analyses suggested that TSG-6 levels correlated positively with optimal fetal metabolic status (cord blood pH). Importantly, TSG-6 expression showed a strong positive correlation with a signature of immunomodulatory microRNAs (miR) (miR-155-5p, miR-21-3p, miR-126-3p). These microRNAs were concurrently downregulated in the hypertensive group; however, because all hypertensive participants received methyldopa, the independent contributions of hypertension and antihypertensive treatment to these changes cannot be distinguished. Gestational hypertension independently reduces the anti-inflammatory TSG-6 in early breast milk. The concomitant reduction in TSG-6 and its correlated microRNA co-expression pattern indicates an altered immunomodulatory profile within the mammary gland during lactation. We hypothesize that these changes might alter the local immunomodulatory properties of milk in the neonatal gastrointestinal tract. However, because no functional in vitro or in vivo assays were performed, this remains a purely theoretical implication that requires future experimental validation.