Epigenetic activation of antiviral sensors and effectors of interferon response pathways during SARS-CoV-2 infection

Recent studies have shown that methylation changes identified in blood cells of COVID-19 patients have a po-tential to be used as biomarkers of SARS-CoV-2 infection outcomes. However, different studies have reported different subsets of epigenetic lesions that stratify patients according to the severity of infection symptoms, and more importantly, the significance of those epigenetic changes in the pathology of the infection is still not clear. We used methylomics and transcriptomics data from the largest so far cohort of COVID-19 patients from four geographically distant populations, to identify casual interactions of blood cells' methylome in pathology of the COVID-19 disease. We identified a subset of methylation changes that is uniformly present in all COVID-19 patients regardless of symptoms. Those changes are not present in patients suffering from upper respiratory tract infections with symptoms similar to COVID-19. Most importantly, the identified epigenetic changes affect the expression of genes involved in interferon response pathways and the expression of those genes differs be-tween patients admitted to intensive care units and only hospitalized. In conclusion, the DNA methylation changes involved in pathophysiology of SARS-CoV-2 infection, which are specific to COVID-19 patients, can not only be utilized as biomarkers in the disease management but also present a potential treatment target

Bortezomib-Induced Epigenetic Alterations in Nerve Cells: Focus on the Mechanisms Contributing to the Peripheral Neuropathy Development

Bortezomib-induced peripheral neuropathy (BiPN) occurs in approximately 40% of patients with multiple myeloma. The induction of severe neuropathy entails the dose reduction or complete elimination of bortezomib (BTZ). Interestingly, discontinuation of BTZ mostly results in a reduction or complete resolution of peripheral neuropathy (PN) symptoms. Therefore, it is likely that the BiPN mechanisms are based on temporary/reversible changes such as epigenetic alterations. In this study, we examined the effect of treating nerve cells, differentiated from the Lund human mesencephalic (dLUHMES) cell line, with several low-dose BTZ (0.15 nM) applications. We showed a significant decrease in global histone H3 acetylation as well as histone H3 lysine 9 acetylation. Moreover, analysis of the genetic microarray showed changes mainly in epigenetic processes related to chromatin rearrangement, chromatin silencing, and gene silencing. GSEA analysis revealed three interesting signaling pathways (SIRT1, B-WICH and, b-Catenin) that may play a pivotal role in PN development. We also performed an analysis of the miRNA microarray which showed the interactions of miR-6810-5p with the genes MSN, FOXM1, TSPAN9, and SLC1A5, which are directly involved in neuroprotective processes, neuronal differentiation, and signal transduction. The study confirmed the existence of BTZ-induced complex epigenetic alterations in nerve cells. However, further studies are necessary to assess the reversibility of epigenetic changes and their potential impact on the induction/resolution of PN

Potencjał wysp Langerhansa do zmian strukturalno-funkcjonalnych podczas ciąży w modelu mysim

Okres ciąży wymaga szeregu adaptacji, które pozwalają na poprawny podział dostępnych zasobów pomiędzy matką a płodem, niezbędny zarówno do wewnątrzmacicznego wzrostu i zaspokojenia bieżących potrzeb matki, jak i do późniejszej laktacji. W modelu mysim, ciężarna samica podlega istotnym zmianom fizjologicznym, które pozwalają na utrzymanie macicy z płodami, stanowiące nawet 20-30% jej masy ciała. Obserwowane jest zwiększenie wagi organów reprodukcyjnych, serca, wątroby i trzustki które, połączone ze zmianami funkcjonalnymi i strukturalnymi, pozwalają sprostać metabolicznym wyzwaniom ciąży. Celem prezentowanej pracy była próba odpowiedź na pytanie, czy następują zmian cytoarchitektury i funkcjonalne wysp trzustkowych w ciąży i wraz z jej progresją, szczególnie w końcowej 1/3 okresu ciąży, kiedy wymogi płodu są największe, a zdolność matki do przekazywania zasobów staje się ograniczona (dzień 16 i dzień 19 z 20-dniowego okresu ciąży). W ramach eksperymentów przeprowadzono porównanie ciężarnych samic (dzień 16 i 19 ciąży) z dziewiczymi samicami i z męską grupą kontrolną. W ramach eksperymentów zastosowano techniki barwienia immunofluorescencyjnego, wykorzystując zarówno mikroskop fluorescencyjny, jak i konfokalny. W dalszych etapach badań zoptymalizowano rozbudowaną procedurę chirurgiczną izolacji wysp Langerhansa z ciężarnych samic, jak również biochemiczne techniki analizy funkcjonalnej in vitro izolowanych wysp. Wszystkie analizowane parametry, w tym wbudowywanie BrdU do DNA, pozytywne barwienie immunofluorescencyjne komórek zawierających aktywna kaspazę-3, występowanie niewielkich wysp trzustkowych oraz pomiar in vitro stymulowanej glukozą sekrecji insuliny wskazują na 16 dzień ciąży, jako czas maksymalnej zdolności do zmian strukturalno-funkcjonalnych wysp Langerhansa, w porównaniu do dziewiczych samic i grupy samców. W dniu 19, przed porodem, nie wykazano proliferacji komórek β, obserwowano zmniejszoną liczbę komórek zawierających aktywną kaspazę-3, a stymulowana glukozą sekrecja insuliny zmniejszyła się do ok. 50% w stosunku do obserwowanej w dniu 16. U dziewiczych samic, podobnie jak u samic w dniu 19 ciąży nie zaobserwowano wbudowywania BrdU do DNA, a poziom aktywnej kaspazy-3 był najniższy ze wszystkich grup samic. Podobnie nie zaobserwowano oznak zmiany struktury wysp Langerhansa u samców, co potwierdza ciążozależny charakter obserwowanych zmian. Podsumowując, w ramach projektu wykazano szereg adaptacji związanych z homeostazą glukozy, które zachodzą poprzez zmiany funkcjonalne i strukturalne wysp trzustkowych.Gestational adaptations are required to achieve the appropriate distribution of resources between the mother and fetus that is optimal for both intrauterine growth and maternal requirements to support the pregnancy and lactation. In mice, the pregnant dam undergoes major physiological changes which enable her to support a gravid uterus that accounts for 20-30% of her mass at term. There is an increase in the weight of the dam’s reproductive tissues, heart, liver and pancreas with potential functional and structural adaptations which help to meet metabolic challenge of pregnancy. The aim of this project was to determine whether the cytoarchitecture and remodeling potential of pancreas islets are affected by pregnancy in the mouse, adding to that analysis of the changes in the function of β-cells with gestational age. This study focused on the terminal 1/3rd of pregnancy, when fetal demand for resources are the greatest and maternal ability to supply becomes constrained (studied days 16 and 19 of pregnancy, term=20 days). It involved the comparison of pregnant females to virgin female counterparts and male mice. The project employed immunostaining, using both, fluorescent and confocal microscopy techniques, the optimization of the surgical technique for islets isolation from female mice and biochemical functional assays of β-cells in vitro. All analyzed parameters as BrdU incorporation, caspase-3-positive cell immunostaining, small islets occurrence and glucose stimulated insulin secretion in vitro indicate day 16 gestational age as the time of maximal remodeling capacity of islets of Langerhans as compared to other analyzed conditions. Near term, at day 19 gestational age, no β-cell proliferation was detected, less of caspase-3 positive cells were observed and insulin index from glucose stimulated insulin secretion was reduced to ~50% of the day 16 gestational age value. Conversely, in virgin counterpart there was no BrdU incorporation detected as on day 19 gestational age and the lowest value of caspase-3 positive cells from all female conditions. Similarly, there was no β-cells remodeling observed in the male pancreas, confirming the pregnancy-dependent character of the observed changes. Overall the project shows that, during mouse pregnancy, there are major adaptations in glucose homeostasis by means of cytoarchitectural and functional changes in pancreas

Peripheral neuropathy in patients with multiple myeloma: molecular effects of bortezomib

Multiple myeloma (MM) is a B cell neoplasm characterized by uncontrolled growth of malignant plasma cells within the bone marrow. The introduction of new treatment regimens and medicinal substances, particularly proteasome inhibitors (e.g. bortezomib or carfilzomib) and immunomodulatory drugs (e.g. lenalidomide, pomalidomide, and monoclonal antibodies), have radically changed MM therapy by improving the response rate and progression-free survival. However, these potentially effective drugs are associated with a number of side effects, the most serious of which include peripheral neuropathy, which appears in 40% of MM patients with bortezomib treatment and up to 70% with thalidomide treatment during long-term exposure. Usually, symptoms of neuropathy disappear after drug discontinuation or dose reduction. However, as a result, the effectiveness of the treatment is lowered and survival time is reduced. The pathogenesis of chemotherapy-induced peripheral neuropathy  is not fully understood. Current research focuses on areas such as the change in the expression of genes responsible for the proper functioning of the nervous system, neuroprotective protein factors, oxidative stress, pro-inflammatory factors and epigenetic changes (miRNA, DNA methylation or histone acetylation). Thoroughly elucidating the mechanisms responsible for the development of chemotherapy-induced peripheral neuropathy will allow us to reduce/eliminate this side effect and improve quality of life for patients

The Evidence That 25(OH)D3 and VK2 MK-7 Vitamins Influence the Proliferative Potential and Gene Expression Profiles of Multiple Myeloma Cells and the Development of Resistance to Bortezomib

Multiple myeloma (MM) remains an incurable hematological malignancy. Bortezomib (BTZ) is a proteasome inhibitor widely used in MM therapy whose potent activity is often hampered by the development of resistance. The immune system is vital in the pathophysiology of BTZ resistance. Vitamins D (VD) and K (VK) modulate the immune system; therefore, they are potentially beneficial in MM. The aim of the study was to evaluate the effect of BTZ therapy and VD and VK supplementation on the proliferation potential and gene expression profiles of MM cells in terms of the development of BTZ resistance. The U266 MM cell line was incubated three times with BTZ, VD and VK at different timepoints. Then, proliferation assays, RNA sequencing and bioinformatics analysis were performed. We showed BTZ resistance to be mediated by processes related to ATP metabolism and oxidative phosphorylation. The upregulation of genes from the SNORDs family suggests the involvement of epigenetic mechanisms. Supplementation with VD and VK reduced the proliferation of MM cells in both the non-BTZ-resistant and BTZ-resistant phenotypes. VD and VK, by restoring proper metabolism, may have overcome resistance to BTZ in vitro. This observation forms the basis for further clinical trials evaluating VD and VK as potential adjuvant therapies for MM patients

Vitamin D as a Potential Player in Immunologic Control over Multiple Myeloma Cells: Implications for Adjuvant Therapies

Multiple myeloma (MM) is a plasma cell malignancy with multifactorial etiology. One of the underlying mechanisms is immune system dysregulation. Immunotherapy is being widely introduced into various MM treatment protocols. Nevertheless, little is known about boosting the immune system with supportive treatment. Although classical actions of vitamin D (VD) are very well established, their non-classical actions related to the modulation of the immune system in MM are still a subject of ongoing research. In this literature review, we intend to summarize research conducted on VD and MM, both in vitro and in vivo, with particular emphasis on immune system modulation, the induction of the differentiation of malignant MM cells, synergic activity with anti-MM drugs, and MM-associated peripheral neuropathy

Molecular Mechanisms of Bortezomib Action: Novel Evidence for the miRNA−mRNA Interaction Involvement

Bortezomib is an anti-tumor agent, which inhibits 26S proteasome degrading ubiquitinated proteins. While apoptotic transcription-associated activation in response to bortezomib has been suggested, mechanisms related to its influence on post-transcriptional gene silencing mediated regulation by non-coding RNAs remain not fully elucidated. In the present study, we examined changes in global gene and miRNA expression and analyzed the identified miRNA–mRNA interactions after bortezomib exposure in human neuroblastoma cells to define pathways affected by this agent in this type of cells. Cell viability assays were performed to assess cytotoxicity of bortezomib. Global gene and miRNA expression profiles of neuroblastoma cells after 24-h incubation with bortezomib were determined using genome-wide RNA and miRNA microarray technology. Obtained results were then confirmed by qRT-PCR and Western blot. Further bioinformatical analysis was performed to identify affected biological processes and pathways. In total, 719 genes and 28 miRNAs were downregulated, and 319 genes and 61 miRNAs were upregulated in neuroblastoma cells treated with bortezomib. Possible interactions between dysregulated miRNA/mRNA, which could be linked to bortezomib-induced neurotoxicity, affect neurogenesis, cellular calcium transport, and neuron death. Bortezomib might exert toxic effects on neuroblastoma cells and regulate miRNA–mRNA interactions influencing vital cellular functions. Further studies on the role of specific miRNA–mRNA interactions are needed to elucidate mechanisms of bortezomib action

Preclinical Evaluation of Long-Term Neuroprotective Effects of BDNF-Engineered Mesenchymal Stromal Cells as Intravitreal Therapy for Chronic Retinal Degeneration in Rd6 Mutant Mice

This study aimed to investigate whether the transplantation of genetically engineered bone marrow-derived mesenchymal stromal cells (MSCs) to overexpress brain-derived neurotrophic factor (BDNF) could rescue the chronic degenerative process of slow retinal degeneration in the rd6 (retinal degeneration 6) mouse model and sought to identify the potential underlying mechanisms. Rd6 mice were subjected to the intravitreal injection of lentivirally modified MSC-BDNF or unmodified MSC or saline. In vivo morphology, electrophysiological retinal function (ERG), and the expression of apoptosis-related genes, as well as BDNF and its receptor (TrkB), were assessed in retinas collected at 28 days and three months after transplantation. We observed that cells survived for at least three months after transplantation. MSC-BDNF preferentially integrated into the outer retinal layers and considerably rescued damaged retinal cells, as evaluated by ERG and immunofluorescence staining. Additionally, compared with controls, the therapy with MSC-BDNF was associated with the induction of molecular changes related to anti-apoptotic signaling. In conclusion, BDNF overexpression observed in retinas after MSC-BDNF treatment could enhance the neuroprotective properties of transplanted autologous MSCs alone in the chronically degenerated retina. This research provides evidence for the long-term efficacy of genetically-modified MSC and may represent a strategy for treating various forms of degenerative retinopathies in the future

Adiponectin, Leptin and Resistin in Patients with Psoriasis

Psoriasis is a common chronic, inflammatory skin disease characterised by keratinocyte hyperproliferation, parakeratosis, and T-cell infiltration. Adipose tissue has an endocrine function, producing an abundance of cytokines and adipokines. It has also been described that the major adipokines, leptin, resistin, and adiponectin, may be involved in the pathogenesis of psoriasis. The aim of the study was to examine the plasma levels of adiponectin, leptin, and resistin in patients with psoriasis and their correlations with disease activity parameters: Psoriasis Activity Severity Index (PASI), Dermatology Life Quality Index (DLQI), and Body Surface Area (BSA) index, as well as selected clinical parameters. The study included 53 patients with the plaque type and 31 healthy controls. The plasma concentrations of adiponectin were significantly lower in patients with psoriasis (p p = 0.2). The plasma concentrations of resistin were significantly increased in patients with psoriasis compared to healthy controls (p = 0.02). There were no statistically significant correlations between adiponectin and leptin plasma concentrations and values of PASI, DLQI, and BSA. The resistin plasma concentrations correlated significantly with DLQI values. Additionally, we examined the correlations between adiponectin, leptin, and resistin plasma concentrations, and selected clinical parameters. Plasma concentrations of adiponectin correlated significantly with CRP values and ALT values. Leptin plasma concentrations correlated significantly with creatinine values. The results of our study confirm the role of adiponectin, leptin, and resistin in the pathogenesis of psoriasis

The Clinical Importance of IL-6, IL-8, and TNF-α in Patients with Ovarian Carcinoma and Benign Cystic Lesions

The exact pathogenesis and influence of various cytokines in patients with ovarian lesions remains unclear. Hence, this study aimed to investigate whether IL-6, IL-8, and TNF-α could be considered as new useful markers for diagnosis of ovarian cancer. 63 women diagnosed with ovarian cancer (OC) and 53 patients with benign ovarian cystic (BOC) lesions were included in this study. Serum levels of IL-6, IL-8, and TNF-α were measured using ELISA. Statistical comparisons were made using the Mann–Whitney U test and all correlations were evaluated by Spearman’s ranks. The serum IL-8 and TNF-α concentration measured in the OC Group was significantly higher than in the BOC Group (p < 0.05). The cutoff level of IL-8 and TNF-α in the serum was set at 4.09 ng/mL and 2.63 ng/mL, respectively, with the sensitivity and specificity of 70% and 96% for IL-8 and 85.7% and 79.3% for TNF-α (p < 0.0001). These results suggest that IL-8 and TNF-α are useful biomarkers for predicting the malignant character of lesions of the ovary. The present study highlighted the importance of measuring the cytokines such as IL-8 and TNF-α in patients with ovarian lesions in predicting the clinical outcome