Regulation Of Cell Function Research Articles

Nicotine induces macrophage pyroptosis via LINC01272/miR-515/KLF6 axis

Nicotine contributes to the causation of atherosclerosis, which the prominent cellular components are macrophages. Long non-coding RNAs (lncRNAs) play an important role in regulating cell functions such as cell proliferation, differentiation and programmed death. However, the function and mechanism of lncRNAs in nicotine-induced macrophage pyroptosis has not been reported. We screened the deferentially expressed lncRNAs of human carotid artery plaque (GSE97210) and verified them in nicotine-induced pyroptosis of macrophages. Results showed only LINC01272 was up-regulated in a dose-dependent manner in macrophages. The immunofluorescence staining result confirmed that interfering LINC01272 inhibited nicotine-induced macrophage pyroptosis. Through bioinformatics analysis, dual luciferase reporter gene assay and qPCR, we identified miR-515 was significantly negatively correlated with the expression of LINC01272, and KLF6 is the target gene of miR-515. Furthermore, our results demonstrated that LINC01272/miR-515/KLF6 axis meditated nicotine-induced macrophage pyroptosis. In addition, in human peripheral blood mononuclear cells of smoking populations, the expression of GSDMD-N, NLRP3, LINC01272 and KLF6 was significantly increased, while the level of miR-515 was reduced. This study confirmed that nicotine increases the expression of LINC01272 to competitively bind with miR-515 in macrophages, reducing the inhibitory effect of miR-515 on its target gene KLF6, which ultimately induces macrophage pyroptosis.

Bariatric Surgery Alters the Postprandial Recovery From Hypoglycemia, Mediated by Cholinergic Signal.

Rerouted gut after Roux-en-Y gastric bypass (GB) and, to a lesser degree, after sleeve gastrectomy (SG) leads to larger glucose excursion and lower nadir glucose, predisposing individuals to hypoglycemia. Despite prandial hyperglucagonemia, endogenous glucose production response to hypoglycemia is reduced after GB or SG. Parasympathetic nervous system (PNS) activity plays a key role in regulation of glucose kinetics and islet cell function. We examined the effect of acute PNS blockade on counterregulatory glucose and islet cell response to meal ingestion during insulin-induced hypoglycemia among GB, SG, and control subjects who had not had gastric surgery. Our findings demonstrate that cholinergic signal is critical in the recovery from hypoglycemia by meal ingestion in humans who have not had gastric surgery, although prandial PNS-mediated physiologic responses to hypoglycemia are differentially changed by GB and SG.

Transcriptional regulation of dendritic cell development and function.

Dendritic cells (DCs) are sentinel immune cells that form a critical bridge linking the innate and adaptive immune systems. Extensive research addressing the cellular origin and heterogeneity of the DC network has revealed the essential role played by the spatiotemporal activity of key transcription factors. In response to environmental signals DC mature but it is only following the sensing of environmental signals that DC can induce an antigen specific T cell response. Thus, whilst the coordinate action of transcription factors governs DC differentiation, sensing of environmental signals by DC is instrumental in shaping their functional properties. In this review, we provide an overview that focuses on recent advances in understanding the transcriptional networks that regulate the development of the reported DC subsets, shedding light on the function of different DC subsets. Specifically, we discuss the emerging knowledge on the heterogeneity of cDC2s, the ontogeny of pDCs, and the newly described DC subset, DC3. Additionally, we examine critical transcription factors such as IRF8, PU.1, and E2-2 and their regulatory mechanisms and downstream targets. We highlight the complex interplay between these transcription factors, which shape the DC transcriptome and influence their function in response to environmental stimuli. The information presented in this review provides essential insights into the regulation of DC development and function, which might have implications for developing novel therapeutic strategies for immune-related diseases.

Influence of e-waste exposure on DNA damage and DNA methylation in people living near recycling sites.

The association between long-term exposure to e-waste and poor health is well established, but how e-waste exposure affects DNA methylation is understudied. In this study, we measured the DNA damage levels and the alternation of DNA methylation in peripheral blood mononuclear cells (PBMCs) collected from a population exposed to e-waste. The concentration of 28 PCB congeners in the blood samples of e-waste recycling workers was elevated than those of the reference group. DNA damage levels were significantly higher than that of samples from the reference group by detecting the SCGE, CA, and CBMN assays. Eventually, we found that the methylation level of 1233 gene loci was changed in the exposure group. Bioinformatic analysis of differential genes revealed that the hypermethylated genes were enriched in cell component movement and regulation of cell function, and hypomethylated genes were involved in the cellular metabolic process. Among the 30 genes we tested, 14 genes showed a negative correlation between methylation level and expression level. Therefore, e-waste exposure potentially increased the levels of DNA damage and alters DNA methylation, which would likely impact human health.

Regulation of Dendritic Cell Functions by Vitamins as Promising Therapeutic Strategy for Immune System Disorders.

A functional immune system is crucial for a healthy life, protecting from infections, tumors, or autoimmune disorders; these are accomplished by the interaction between various immune cells. Nourishment, particularly micronutrients, are very important components in the immune system balance, therefore this review emphasizes the vitamins (D, E, A, C) and Dendritic cells' subsets due to vitamins' roles in immune processes, especially on dendritic cells' functions, maturation, and cytokine production. Current studies reveal significant benefits related to vitamins, including vitamin E, which can contribute to the control of dendritic cells' function and maturation. Furthermore, vitamin D plays an immunoregulatory and anti-inflammatory role in the immune system. Metabolite of vitamin A which is called retinoic acid leads to T cells' differentiation to T helper 1 or T helper 17, so low levels of this vitamin exacerbate the menace of infectious diseases, and vitamin C has anti-oxidant effects on dendritic cells and modulate their activation and differentiation program. Additionally, the correlation between the amount of vitamin and the occurrence or progression of allergic diseases and autoimmunity disorders is discussed according to the results of previous studies.

MicroRNA-29 differentially mediates preeclampsia-dysregulated cellular responses to cytokines in female and male fetal endothelial cells.

Preeclampsia (PE) differentially impairs female and male fetal endothelial cell function, which is associated with an increased risk of adult-onset cardiovascular disorders in children born to mothers with PE. However, the underlying mechanisms are poorly defined. We hypothesize that dysregulation of microRNA-29a-3p and 29c-3p (miR-29a/c-3p) in PE disturbs gene expression and cellular responses to cytokines in fetal endothelial cells in a fetal sex-dependent manner. RT-qPCR analysis of miR-29a/c-3p was performed on female and male unpassaged (P0) human umbilical vein endothelial cells (HUVECs) from normotensive (NT) pregnancies and PE. Bioinformatic analysis of an RNA-seq dataset was performed to identify PE-dysregulated miR-29a/c-3p target genes in female and male P0-HUVECs. Gain- and loss-of-function assays were conducted to determine the effects of miR-29a/c-3p on endothelial monolayer integrity and proliferation in response to transforming growth factor-β1 (TGFβ1) and tumour necrosis factor-α (TNFα) in NT and PE HUVECs at passage 1. We observed that PE downregulated miR-29a/c-3p in male and female P0-HUVECs. PE dysregulated significantly more miR-29a/c-3p target genes in female vs. male P0-HUVECs. Many of these PE-differentially dysregulated miR-29a/c-3p target genes are associated with critical cardiovascular diseases and endothelial function. We further demonstrated that miR-29a/c-3p knockdown specifically recovered the PE-abolished TGFβ1-induced strengthening of endothelial monolayer integrity in female HUVECs, while miR-29a/c-3p overexpression specifically enhanced the TNFα-promoted cell proliferation in male PE HUVECs. In conclusion, PE downregulates miR-29a/c-3p expression and differentially dysregulates miR-29a/c-3p target genes associated with cardiovascular diseases and endothelial function in female and male fetal endothelial cells, possibly contributing to the fetal sex-specific endothelial dysfunction observed in PE. KEY POINTS: Preeclampsia differentially impairs female and male fetal endothelial cell function in responses to cytokines. Pro-inflammatory cytokines are elevated in maternal circulation during pregnancy in preeclampsia. MicroRNAs are critical regulators of endothelial cell function during pregnancy. We have previously reported that preeclampsia downregulated microRNA-29a-3p and 29c-3p (miR-29a/c-3p) in primary fetal endothelial cells. However, it is unknown if PE differentially dysregulates the expression of miR-29a/c-3p in female and male fetal endothelial cells. We show that preeclampsia downregulates miR-29a/c-3p in male and female HUVECs and preeclampsia dysregulates cardiovascular disease- and endothelial function-associated miR-29a/c-3p target genes in HUVECs in a fetal sex-specific manner. MiR-29a/c-3p differentially mediate cell responses to cytokines in female and male fetal endothelial cells from preeclampsia. We have revealed fetal sex-specific dysregulation of miR-29a/c-3p target genes in fetal endothelial cells from preeclampsia. This differential dysregulation may contribute to fetal sex-specific endothelial dysfunction in offspring born to preeclamptic mothers.

The impact of high-fat diet-induced oxidative stress on micro RNA’s in various tissues

Stress is the body’s reaction to any kind of injury or danger. It is linked to the production of oxidative free radicals, which are responsible for a variety of acute, chronic, and potentially fatal illnesses and diseases. Free radicals, due to their extreme reactivity, can harm or even kill cells. A High-Fat Diet (HFD) causes “oxidative stress”, which is characterized by an increase in the body’s generation of Reactive Oxygen Species (ROS) as a result of higher levels of triglycerides and Free Fatty Acids (FFA). HFD-induced oxidative stress alters cellular function by affecting transcriptional factors and mitochondrial enzymes (synthesis/inhibition). ROS and FFA damage the receptors of the epithelium, resulting in epithelial damage that impairs cellular function. ROS levels can harm cells by altering the expression of microRNA (miRNA), a sign of RNA damage. MiRNAs are non-coding RNAs found in animals, plants, and some viruses that play a role in the post-transcriptional regulation of gene expression. These three pathways—RNA cleavage, RNA destabilization, and RNA translation into proteins— all play a role in mRNA expression. The miRNA regulates the up- and downregulation of mRNA expression for cellular function, enzyme synthesis, and receptor modulation. MiRNA regulates cell function by maintaining the balance between cellular ROS levels and cellular damage.

Infection Meets Inflammation: N6-Methyladenosine, an Internal Messenger RNA Modification as a Tool for Pharmacological Regulation of Host-Pathogen Interactions.

The significance of internal mRNA modifications for the modulation of transcript stability, for regulation of nuclear export and translation efficiency, and their role in suppressing innate immunity is well documented. Over the years, the molecular complexes involved in the dynamic regulation of the most prevalent modifications have been characterized-we have a growing understanding of how each modification is set and erased, where it is placed, and in response to what cues. Remarkably, internal mRNA modifications, such as methylation, are emerging as an additional layer of regulation of immune cell homeostasis, differentiation, and function. A fascinating recent development is the investigation into the internal modifications of host/pathogen RNA, specifically N6-methyladenosine (m6A), its abundance and distribution during infection, and its role in disease pathogenesis and in shaping host immune responses. Low molecular weight compounds that target RNA-modifying enzymes have shown promising results in vitro and in animal models of different cancers and are expanding the tool-box in immuno-oncology. Excitingly, such modulators of host mRNA methyltransferase or demethylase activity hold profound implications for the development of new broad-spectrum therapeutic agents for infectious diseases as well. This review describes the newly uncovered role of internal mRNA modification in infection and in shaping the function of the immune system in response to invading pathogens. We will also discuss its potential as a therapeutic target and identify pitfalls that need to be overcome if it is to be effectively leveraged against infectious agents.

Pregnancy-specific beta-1-glycoprotein 1-enriched exosomes are involved in the regulation of vascular endothelial cell function during pregnancy

IntroductionPregnancy is a dynamic time period associated with significant physiological changes in the cardiovascular system. It is well known that during pregnancy, the placenta secretes a variety of molecular signals, including exosomes, into the maternal circulation to adapt to increased blood volume and maintain blood pressure at normotensive levels. MethodsIn the present study, we compared the effects of exosomes derived from the peripheral blood serum of nonpregnant women (NP-Exo) and pregnant women with uncomplicated pregnancy (P-Exo) on endothelial cell function. We also analyzed the proteomic profiles of these two groups of exosomes and the molecular mechanisms underlying the effect of exosome cargoes on vascular endothelial cell function. ResultsWe found that P-Exo were positively involved in regulating the function of human umbilical vein endothelial cell (HUVEC) and promoting the release of nitric oxide (NO). Furthermore, we revealed that trophoblast-derived pregnancy-specific beta-1-glycoprotein 1 (PSG1)-enriched exosomes treatment induced the promotion of HUVEC proliferation and migration as well as the release of NO. In addition, we found that P-Exo maintained blood pressure at normal levels in mice. DiscussionThese results suggested that PSG1-enriched exosomes derived from maternal peripheral blood regulate the function of vascular endothelial cells and play an important role in maintaining maternal blood pressure during pregnancy.

In Situ Activation of the Receptor Aggregation for Cell Apoptosis by an AI-Au Intelligent Nanomachine via Tumor Extracellular Acidity.

Polyvalent ligand-induced cell receptor aggregation is closely related to cell behavior regulation. At present, most of the means to induce receptor aggregation rely on external stimuli such as light, heat, and magnetic fields, which may cause side effects to normal cells. How to achieve receptor aggregation on the cancer cell surface to achieve cell apoptosis selectively is still a challenge. Therefore, by taking advantage of the unique property of cancer cells' slightly acidic microenvironment, an easy-to-use apoptosis-inducing mode for the in situ activation of cell surface nucleolin clustering has been developed, which not only opened a new channel for nucleolin receptor aggregation to regulate cell function and further development but also avoided damage to normal cells, providing a new strategy for tumor treatment. Dual functional ssDNA (AS1411 aptamer and pH-responsive I-strand sequence) was modified on the surface of gold nanoparticles (AuNPs) to fabricate AI-Au intelligent nanomachines. Then, the specific binding on cancer cells and aggregation of the nucleolin receptors can be achieved via the formation of an i-Motif structure among adjacent AuNPs under the acidic microenvironment. The result showed that AI-Au nanomachines mediated nucleolin cross-linking on the cell surface, resulting in a cytotoxic effect of approximately 60%. Experiments such as calcein-AM/PI staining, nuclear dye staining, and flow cytometry demonstrated that cell apoptosis became more evident with the increase of acidity under the cell surface microenvironment. Immunofluorescence imaging further confirmed the Cyt-c/caspase-3 apoptosis pathway induced by AI-Au nanomachines. The proposed strategy used for specific cancer cell apoptosis by the in situ activation of tumor cell membrane receptor aggregation is inexpensive and simple to use, which not only provides a new means of nucleolin receptor aggregation for regulating cell function but also offers a new strategy for tumor treatment with reduced side effect to normal cells. This work is significant for comprehending the ligand-induced receptor aggregation process and can lead to the development of a promising anticancer drug.

1628-P: Role of CD38 in the Regulation of Muscle Stem Cell Functions in Aged Mice

Skeletal muscle repair and regeneration is a complex process and is known to be delayed with aging, diabetes and obesity. This is in part due to infiltration of inflammatory macrophages which can delay the recovery process. Both macrophages and satellite cells require nicotinamide adenine dinucleotide (NAD+) for their biological functions. Aging-induced decline in NAD+ have been shown to worsen insulin resistance and muscle dysfunction. CD38 is an ectoenzyme that degrades NAD+ and is increased in inflammatory macrophages in aging, obesity and other insulin-resistant states. However, how inhibition of CD38 affects satellite cell functions in aged or obese mice, remains unknown. Here, we utilized aged CD38 knockout mice to assess the role of this protein and NAD+ in the recovery process in injured muscle. CD38 KO mice were found to be resistant to diet-induced obesity and had improved insulin sensitivity. Our data show that genetic deletion of CD38 in aged mice increased NAD+ levels and thus enhanced muscle stem/satellite cells differentiation and promote myogenesis. Deletion of CD38 also resulted in an enhanced number of regenerating myofibers. Furthermore, we found that deletion of CD38 significantly reduces fibrosis of muscle and also improved limb grip strength. Pharmacological inhibition of CD38 in C2C12 myoblasts and ex-vivo primary myoblasts significantly increased NAD+ levels and also enhanced the differentiation of satellite cells into myoblasts and myofibers. We, therefore, examined the rescue effects of 78c (CD38i) on human recombinant CD38 (hCD38) inhibition of muscle stem cells. Administration of hCD38 inhibited expression of MyoD and MyoG in primary myoblasts, while this effect was rescued through 78c treatment. Taken together, these data demonstrate that restoration of NAD+ levels either through genetic deletion/pharmacological inhibition of CD38 can improve muscle stem cell functions and muscle regeneration and might promote muscle regeneration in obesity, type 2 diabetes, and aging. Disclosure A.Nawaz: None. K.Yaku: None. I.Jeelani: None. K.Tobe: Other Relationship; MSD K.K., Novo Nordisk Pharma Ltd., Takeda Pharmaceutical Company Limited, Daiichi Sankyo, Mitsubishi Tanabe Pharma Corporation, Suntory Global Corporation, Ltd.,, Taiho Pharmaceutical Co. Ltd., Japan Diabetes Foundation, Japan Association for Diabetes Education and Care. T.Nakagawa: None.

290-LB: Single-Cell Analysis of Endocrine Development Identifies Novel Roles for GLIS3 in Pancreatic ß-Cell Fate

The Krüppel-like zinc finger transcription factor GLIS3 has been identified as a critical regulator of pancreatic beta cell development and function. Mice lacking a functional Glis3 gene fail to produce enough insulin and suffer from severe hyperglycemia soon after birth. This phenotype matches what has been observed in humans with genetic mutations in GLIS3 and who have been diagnosed with permanent neonatal diabetes within the first month of life. Additional and ongoing work has begun to identify some of the transcriptional mechanisms utilized by GLIS3, yet many questions remain unanswered. One such question is whether GLIS3 has roles in endocrine development prior to the formation of functional β-cells. To address this question, I have utilized single cell RNA-sequencing of mouse embryonic pancreas, at embryonic days (e)15.5 and e18.5, of both wild type (WT) and Glis3 knockout (KO) embryos. Analysis of this data has identified the expected developmental populations, from bipotent progenitor cells to pro-endocrine cells and ultimately hormone positive β-, α-, δ-, and ε-cells. Interestingly, there is little difference in gene expression between the WT and Glis3 KO pancreas in most of the populations observed (pro-endocrine, FEV+, α-/δ-/ε-cells). However, there are significant differences in gene expression between WT β-cells and KO β-cells. Surprisingly, most of the differences are in genes upregulated in the KO cells, with only a small number of genes downregulated. These upregulated genes are normally downregulated over the course of development from FEV+ to β-cells. These data indicate that GLIS3 plays a unique role in developing β-cells in that it primarily represses genes during the course of development but is also critical for the transcriptional activation of specific targets, particularly Ins2. Understanding how GLIS3 regulates β-cell generation, and how GLIS3 itself is regulated, could potentially provide new therapeutic avenues for the treatment of diabetes. Disclosure D. W. Scoville: None. S. Grimm: None. A. M. Jetten: None. Funding National Institutes of Health (Z01-ES-101585)

Biological and Catalytic Properties of Selenoproteins.

Selenocysteine is a catalytic residue at the active site of all selenoenzymes in bacteria and mammals, and it is incorporated into the polypeptide backbone by a co-translational process that relies on the recoding of a UGA termination codon into a serine/selenocysteine codon. The best-characterized selenoproteins from mammalian species and bacteria are discussed with emphasis on their biological function and catalytic mechanisms. A total of 25 genes coding for selenoproteins have been identified in the genome of mammals. Unlike the selenoenzymes of anaerobic bacteria, most mammalian selenoenzymes work as antioxidants and as redox regulators of cell metabolism and functions. Selenoprotein P contains several selenocysteine residues and serves as a selenocysteine reservoir for other selenoproteins in mammals. Although extensively studied, glutathione peroxidases are incompletely understood in terms of local and time-dependent distribution, and regulatory functions. Selenoenzymes take advantage of the nucleophilic reactivity of the selenolate form of selenocysteine. It is used with peroxides and their by-products such as disulfides and sulfoxides, but also with iodine in iodinated phenolic substrates. This results in the formation of Se-X bonds (X = O, S, N, or I) from which a selenenylsulfide intermediate is invariably produced. The initial selenolate group is then recycled by thiol addition. In bacterial glycine reductase and D-proline reductase, an unusual catalytic rupture of selenium-carbon bonds is observed. The exchange of selenium for sulfur in selenoproteins, and information obtained from model reactions, suggest that a generic advantage of selenium compared with sulfur relies on faster kinetics and better reversibility of its oxidation reactions.

Autocrine Proteinase-Activated Receptor (PAR) signaling in PC3 prostate cancer cells.

Proteinase-activated receptors (PARs) are G protein-coupled receptors (GPCRs) activated by limited N-terminal proteolysis. PARs are highly expressed in many cancer cells including PCa and regulate various aspects of tumor growth and metastasis. Specific activators of PARs in different physiological and pathophysiological contexts remain poorly defined. In this study, we examined the androgen independent human prostatic cancer cell line PC3 and find functional expression of PAR1 and PAR2 but not PAR4. Using genetically encoded PAR cleavage biosensors, we show that PC3 cells secrete proteolytic enzymes that cleave PARs and trigger autocrine signaling. CRISPR/Cas9 targeting of PAR1 and PAR2 combined with microarray analysis revealed genes that are regulated through this autocrine signaling mechanism. We find several genes that are known PCa prognostic factors or biomarker to be differentially expressed in the PAR1-KO and PAR2-KO PC3 cells. We further examined PAR1 and PAR2 regulation of PCa cell proliferation and migration and find that absence of PAR1 promotes PC3 cell migration and suppresses cell proliferation while PAR2 deficiency showed opposite effects. Overall, these results demonstrate that autocrine signaling through PARs is an important regulator of PCa cell function.

Integrated Analysis of Single-Cell RNA-Seq and Bulk RNA-Seq Combined with Multiple Machine Learning Identified a Novel Immune Signature in Diabetic Nephropathy.

Increasing evidence suggests that immune modulation contributes to the pathogenesis and progression of diabetic nephropathy (DN). However, the role of immune modulation in DN has not been elucidated. The purpose of this study was to search for potential immune-related therapeutic targets and molecular mechanisms of DN. Gene expression datasets were obtained from the Gene Expression Omnibus (GEO) database. A total of 1793 immune-related genes were acquired from the Immunology Database and Analysis Portal (ImmPort). Weighted gene co-expression network analysis (WGCNA) was performed for GSE142025, and the red and turquoise co-expression modules were found to be key for DN progression. We utilized four machine learning algorithms, namely, random forest (RF), support vector machine (SVM), adaptive boosting (AdaBoost), and k-nearest neighbor (KNN), to evaluate the diagnostic value of hub genes. Immune infiltration patterns were analyzed using the CIBERSORT algorithm, and the correlation between immune cell type abundance and hub gene expression was also investigated. A total of 77 immune-related genes of advanced DN were selected for subsequent analyzes. Functional enrichment analysis showed that the regulation of cytokine-cytokine receptor interactions and immune cell function play a corresponding role in the progression of DN. The final 10 hub genes were identified through multiple datasets. In addition, the expression levels of the identified hub genes were corroborated through a rat model. The RF model exhibited the highest AUC. CIBERSORT analysis and single-cell sequencing analysis revealed changes in immune infiltration patterns between control subjects and DN patients. Several potential drugs to reverse the altered hub genes were identified through the Drug-Gene Interaction database (DGIdb). This pioneering work provided a novel immunological perspective on the progression of DN, identifying key immune-related genes and potential drug targets, thus stimulating future mechanistic research and therapeutic target identification for DN.

Interim results from a phase I/II study of duvelisib PI3Kδγ inhibitor and nivolumab in patients with advanced unresectable melanoma who have progressed on anti-PD1 therapy.

9540 Background: The majority of unresectable melanoma patients eventually progress on first-line checkpoint blockade therapy. Duvelisib is a potent phosphoinositide 3-kinase (PI3K) δ and γ isoform inhibitor approved for relapsed/refractory CLL. PI3Kγ inhibition has been shown to restore anti-PD1 activity in preclinical tumor models through conversion of tumor associated macrophages (TAMs) from an immunosuppressive to a pro-inflammatory state. PI3Kδ inactivation has been associated with impaired T regulatory cell (Treg) function and reduced intratumoral Treg numbers. We report study design and results of a phase I study to evaluate the safety and efficacy of duvelisib and nivolumab in anti-PD1 refractory patients. Methods: Utilizing a modified 3+3 design with escalating doses of duvelisib (15 mg daily, 25 mg daily, and 25 mg BID), patients with unresectable melanoma were treated with nivolumab and duvelisib until unacceptable toxicity or disease progression. Primary endpoints for the phase I portion were maximum tolerated dose (MTD) and recommended phase II dose (RP2D) of duvelisib. Secondary endpoints include early and late toxicities and anti-tumor activity based on RECIST v1.1. Exploratory endpoints were to evaluate dose-dependent changes in the immune cell populations of peripheral blood and tumor tissue with duvelisib and nivolumab. Results: Ten patients have been enrolled (7 at dose level I, 3 at dose level II) with mean age 64. Two patients had BRAF V600E mutation and were previously treated with BRAF/MEK inhibitors. No dose limiting toxicities (DLT) were observed. Grade 3 or higher adverse events (AEs) attributable to study treatment occurred in 4 patients (40%), including G3 neutropenia (10%), G3 diarrhea (10%), G3 vomiting (10%) and G4 hepatitis (20%). Both instances of hepatitis resolved rapidly with 1-2mg/kg of prednisone. In one patient, hepatitis and nausea/vomiting led to discontinuation of treatment. Out of 6 patients who had follow-up imaging, 4 exhibited progressive disease, one stable disease, and one had partial response of 11 months duration. Given the published data on the regulation of macrophages and Tregs by PI3Kδγ inhibitors, we are utilizing multispectral imaging to measure the spatial interactions of key immune cells within the tumor microenvironment (T cells, B cells, and macrophages) in all matched pre-/post-treatment paraffin-embedded tissues, for which results will be presented. Conclusions: The combination of duvelisib and nivolumab was well-tolerated in patients with advanced unresectable melanoma refractory to ICI without DLTs to date at 15 and 25mg daily dosages. Significant anti-tumor activity was noted in one patient. Enrollment at dose level II is ongoing. Clinical trial information: NCT04688658 .

3D micropattern force triggers YAP nuclear entry by transport across nuclear pores and modulates stem cells paracrine.

Biophysical cues of the cellular microenvironment tremendously influence cell behavior by mechanotransduction. However, it is still unclear how cells sense and transduce the mechanical signals from 3D geometry to regulate cell function. Here, the mechanotransduction of human mesenchymal stem cells (MSCs) triggered by 3D micropatterns and its effect on the paracrine of MSCs are systematically investigated. Our findings show that 3D micropattern force could influence the spatial reorganization of the cytoskeleton, leading to different local forces which mediate nucleus alteration such as orientation, morphology, expression of Lamin A/C and chromatin condensation. Specifically, in the triangular prism and cuboid micropatterns, the ordered F-actin fibers are distributed over and fully transmit compressive forces to the nucleus, which results in nuclear flattening and stretching of nuclear pores, thus enhancing the nuclear import of YES-associated protein (YAP). Furthermore, the activation of YAP significantly enhances the paracrine of MSCs and upregulates the secretion of angiogenic growth factors. In contrast, the fewer compressive forces on the nucleus in cylinder and cube micropatterns cause less YAP entering the nucleus. The skin repair experiment provides the first in vivo evidence that enhanced MSCs paracrine by 3D geometry significantly promotes tissue regeneration. The current study contributes to understanding the in-depth mechanisms of mechanical signals affecting cell function and provides inspiration for innovative design of biomaterials.

Photobiomodulation in human neuroblastoma under oxidative stress

AbstractBackgroundWith the increase in life expectancy, the number of people with neurodegenerative diseases such as Alzheimer’s Disease (AD) is rising rapidly. Considering the pathways of AD, one of its fundamental processes is oxidative stress, affecting the central nervous system and cell regeneration. Interestingly, in the last decades, photobiomodulation has shown positive effects on neural cells that are compromised by AD. Studies have demonstrated that LED (light emitting diode) irradiation regulates cell function, stimulating cell proliferation, promoting biomodulation in the cytoskeleton, and reducing apoptosis. To contribute with the elucidation of the importance of photobiomodulation in AD, this study aims to evaluate its neuroprotective effect, comparing different energy density of LED irradiating in SH‐SY5Y cells (human neuroblastoma) under the effect of oxidative stress induced by hydrogen peroxide (H2O2).MethodSH‐SY5Y cells (human neuroblastoma) were cultivated in DMEM/F12 medium (Dulbecco’s modified Eagles medium ‐ Gibco), supplemented with 10% fetal bovine serum and 1% antibiotic and antimycotic, at 37°C, in an atmosphere of 5% CO2. Cells were plated in 24‐well plates. After cell adhesion, cells were irradiated with LED at 660 nm (LED Biopdi/Irrad‐Led5), at energy densities of 1 J/cm2, 3 J/cm2, 5 J/cm2 and 10 J/ cm2. 24 h after irradiation, oxidative stress was induced with 200 μM H2O2 for 24h (this parameter had been previously determined by ROSSATO, R. C. et al, 2021).ResultOnce cells are irradiated and stressed, a considerable statistic difference is observed for all energy densities evaluated, compared to the stressed‐only group (H2O2). Importantly, the energy density of 3 J/cm2 was the lowest one that obtained better statistical increase in mithocondrial activity (p < 0.0001).ConclusionIt was possible to determine and compare the energy densities of LED irradiation to promote cellular photobiomodulation, inducing mithocondrial activity on neubroblastoma. Importantly, this study demonstrated the neuroprotective effect of LED in an in vitro model of AD oxidative stress, approaching novel concepts and a preventive tool.ROSSATO, R. C. et al. Neuroprotective effects of taurine on SH‐SY5Y cells under hydrocortisone induced stress. Research, Society and Development, v. 10, n. 9, p. e55510918426‐e55510918426, 2021

Relationship between obesity, insulin resistance and cell membrane properties

Introduction and aim. The obesity is one of the greatest public health problems in developing countries and it is a triggering factor for diabetes associated with insulin resistance. The importance of cell membrane lipids as essential regulators of insulin resistance, since changes in the dynamic properties of the cell membrane (e.g., membrane fluidity), could be one of the events by which obesity affects insulin sensitivity. Thus, the insulin resistance may not only be a cause but also a consequence of lipid disorders such as dyslipidemia and/or cell membrane phospholipid composition change. The modification of plasma membrane lipid composition can change membrane biophysical properties and thus influencing protein-lipid interactions, enzymatic activity and regulation of surface receptors. Alterations in the lipid composition modify the fluidity of plasma membranes and the expression of membrane functions, such as receptor binding and enzyme activities. This review summarizes the current knowledge on the effects of the modulation of plasma membrane lipid composition and membrane fluidity in the functionality of membrane proteins involved in insulin activity, including the insulin receptor, glucose transport and Na + /K + ATPase and, in turn, the key features of the metabolic syndrome. Material and methods. References for that article were found through PubMed and Google Scholar, using terms: “obesity”, “insulin resistance” and “membrane properties”. The research was limited to abstracts and available full-text articles. Analysis of the literature. There is a strong relationship between dietary lipids, membrane lipid profiles and insulin resistance. The changes in the dynamic properties of the cell membrane (e.g., membrane fluidity), could be one of the events by which obesity affects insulin sensitivity. The modification of plasma membrane lipid composition can change membrane biophysical properties and thus influencing protein–lipid interactions, enzymatic activity, and regulation of surface receptors. Modifications of membrane phospholipid composition could have a role in the insulin action by altering membrane fluidity and, as a consequence, the insulin signaling pathway. Conclusion. As conclusion the membrane-lipid therapy approach can be used to treat important pathologies such as obesity and many others diseases such as : cancer, cardiovascular pathologies, neurodegenerative processes, obesity, metabolic disorders, inflammation, and infectious and autoimmune diseases. This pharmacological strategy aims to regulate cell functions by influencing lipid organization and membrane fluidity, inducing a concomitant modulation of membrane protein localization and activity which might serve to reverse the pathological state. Through this review we suggest an in-depth analysis of the membrane lipid therapy field, especially its molecular bases and its relevance to the development of innovative therapeutic approaches.

Novel approaches for allergen-specific immunotherapy—An overview

Allergen-specific immunotherapy (AIT) is an allergen-specific treatment for people with IgE-related allergies. Allergen-specific immunotherapy (AIT) is used to treat allergic disorders when symptoms persist despite medication and allergen avoidance. The therapy is presumed effective if it reduces the use of medications, improves the quality of life even after discontinuation of treatment, as well as prevents the conversion of one type of allergy to the other and the development of new sensitization. The allergen-specific immunotherapeutic agents can be administered sublingually, subcutaneously, or through some other routes, such as intra-lymphatically and epicutaneously to induce allergen tolerance by modifying immune responses (innate and adaptive). The primary mechanism of AIT is the induction of functional regulatory cells, such as regulatory T cells, follicular T cells, B cells, dendritic cells, innate lymphoid cells, and natural killer cells, which results in the control of the functions of type 2 inflammatory cells. However, there are several downsides to AIT, including the contentious treatment period resulting in high cost, systemic allergic reactions, and the lack of a biomarker for forecasting treatment responders. Vaccine adjuvants, adjunctive therapies, and novel vaccine technologies are currently being researched to address the issues associated with AIT. This article focuses on defined molecular approaches for improving the potential of specific immunotherapy that use recombinant allergen derivatives, allergen-derived peptides, virus-coupled allergens, nanoparticles, and specific adjuvants.