ABSTRACT
Evidence abounds that gut microbiome components are associated with sex disparities in the immune system. However, it remains unclear whether the observed sex disparity in asthma incidence is associated with sex-dependent differences in immune-modulating gut microbiota, and/or its influence on allergic airway inflammatory processes. Using a mouse model of house dust mite (HDM)-induced allergic inflammation and the four core genotypes (FCGs) model, we have previously reported sex differences in lung inflammatory phenotypes. Here, we investigated associations of gut microbiomes with these phenotypes by challenging FCG mice [mouse with female sex chromosome and male gonad (XXM), mouse with female sex chromosome and female gonad (XXF), mouse with male sex chromosome and male gonad (XYM), and mouse with male sex chromosome and female gonad (XYF); n = 7/group] with HDM (25 µg) or PBS intranasally for 5 wk and collecting fecal samples. We extracted fecal DNA and analyzed the 16S microbiome via Targeted Metagenomic Sequencing. We compared α and ß diversity across genotypes and assessed the Firmicutes/Bacteroidetes (F/B) ratio. When comparing baseline and after exposure for the FCG, we found that the gut F/B ratio was only increased in the XXM genotype. We also found that α diversity was significantly increased in all FCG mice upon HDM challenge, with the highest increase in the XXF, and the lowest in the XXM genotypes. Similarly, ß diversity of the microbial community was also affected by challenge in a gonad- and chromosome-dependent manner. In summary, our results indicated that HDM treatment, gonads, and sex chromosomes significantly influence the gut microbial community composition. We concluded that allergic lung inflammation may be affected by the gut microbiome in a sex-dependent manner involving both hormonal and genetic influences.NEW & NOTEWORTHY Recently, the gut microbiome and its role in chronic respiratory disease have been the subject of extensive research and the establishment of its involvement in immune functions. Using the FCG mouse model, our findings revealed the influence of gonads and sex chromosomes on the microbial community structure before and after exposure to HDM. Our data provide a potential new avenue to better understand mediators of sex disparities associated with allergic airway inflammation.
Subject(s)
Disease Models, Animal , Gastrointestinal Microbiome , Animals , Gastrointestinal Microbiome/genetics , Female , Male , Mice , Sex Chromosomes/genetics , Asthma/immunology , Asthma/microbiology , Asthma/genetics , Pyroglyphidae/immunology , Inflammation/genetics , Inflammation/immunology , Inflammation/microbiology , Genotype , Gonads/microbiology , Hypersensitivity/immunology , Hypersensitivity/microbiology , Hypersensitivity/genetics , Sex CharacteristicsABSTRACT
Sex differences in allergic inflammation have been reported, but the mechanisms underlying these differences remain unknown. Contributions of both sex hormones and sex-related genes to these mechanisms have been previously suggested in clinical and animal studies. Here, Four-Core Genotypes (FCG) mouse model was used to study the inflammatory response to house dust mite (HDM) challenge and identify differentially expressed genes (DEGs) and regulatory pathways in lung tissue. Briefly, adult mice (8-10 wk old) of the FCG (XXM, XXF, XYM, XYF) were challenged intranasally with 25 µg of HDM or vehicle (PBS-control group) 5 days/wk for 5 wk (n = 3/10 group). At 72 h after the last exposure, we analyzed the eosinophils and neutrophils in the bronchoalveolar lavage (BAL) of FCG mice. We extracted lung tissue and determined DEGs using Templated Oligo-Sequencing (TempO-Seq). DEG analysis was performed using the DESeq2 package and gene enrichment analysis was done using Ingenuity Pathway Analysis. A total of 2,863 DEGs were identified in the FCG. Results revealed increased eosinophilia and neutrophilia in the HDM-treated group with the most significantly expressed genes in XYF phenotype and a predominant effect of female hormones vs. chromosomes. Regardless of the sex hormones, mice with female chromosomes had more downregulated genes in the HDM group but this was reversed in the control group. Interestingly, genes associated with inflammatory responses were overrepresented in the XXM and XYF genotypes treated with HDM. Sex hormones and chromosomes contribute to inflammatory responses to HDM challenge, with female hormones exerting a predominant effect mediated by inflammatory DEGs.NEW & NOTEWORTHY Gene expression profiling helps to provide deep insight into the global view of disease-related mechanisms and responses to therapy. Using the Four-Core Genotype mouse model, our findings revealed the influence of sex hormones and sex chromosomes in the gene expression of lungs exposed to an aeroallergen (House Dust Mite) and identified sex-specific pathways to better understand sex disparities associated with allergic airway inflammation.
Subject(s)
Allergens , Lung , Female , Mice , Male , Animals , Allergens/metabolism , Lung/metabolism , Disease Models, Animal , Gene Expression Profiling , Pyroglyphidae , Inflammation/genetics , Inflammation/metabolism , Gonadal Steroid Hormones/metabolism , Genotype , Gene Expression , Hormones/metabolism , Bronchoalveolar Lavage FluidABSTRACT
Epigenetic alterations such as dysregulation of miRNAs have been reported to play important roles in interactions between genetic and environmental factors. In this study, we tested the hypothesis that induction of lung inflammation by inhaled allergens triggers a sex-specific miRNA regulation that is dependent on chromosome complement and hormonal milieu. We challenged the four core genotypes (FCGs) model through intranasal sensitization with a house dust mite (HDM) solution (or PBS as a control) for 5 wk. The FCG model allows four combinations of gonads and sex chromosomes: 1) XX mice with ovaries (XXF), 2) XY mice with testes (XYM), 3) XX mice with testes (XXM), and 4) XY mice with ovaries (XYF). Following the challenge (n = 5-7/group), we assessed the expression of 84 inflammatory miRNAs in lung tissue using a PCR array and cytokine levels in bronchoalveolar lavage fluid (BAL) by a multiplex protein assay (n = 4-7 animals/group). Our results showed higher levels of the chemokine KC (an Il-8 homolog) and IL-7 in BAL from XYF mice challenged with HDM. In addition, IL-17A was significantly higher in BAL from both XXF and XYF mice. A three-way interaction among treatment, gonads, and sex chromosome revealed 60 of 64 miRNAs that differed in expression depending on genotype; XXF, XXM, XYF, and XYM mice had 45, 32, 4, and 52 differentially expressed miRNAs, respectively. Regulatory networks of miRNAs identified in this study were implicated in pathways associated with asthma. Female gonadal hormonal effects may alter miRNA expression and contribute to the higher susceptibility of females to asthma.NEW & NOTEWORTHY miRNAs play important roles in regulating gene and environmental interactions. However, their role in mediating sex differences in allergic responses and lung diseases has not been elucidated. Our study used a targeted omics approach to characterize the contributions of gonadal hormones and chromosomal components to lung responses to an allergen challenge. Our results point to the influence of sex hormones in miRNA expression and proinflammatory markers in allergic airway inflammation.
Subject(s)
Asthma , MicroRNAs , Female , Animals , Mice , Male , Cytokines/genetics , MicroRNAs/genetics , MicroRNAs/metabolism , Lung/metabolism , Sex Chromosomes/genetics , Sex Chromosomes/metabolism , Asthma/genetics , Asthma/metabolism , Inflammation/genetics , Inflammation/metabolism , Bronchoalveolar Lavage Fluid , Gonadal Hormones/genetics , Gonadal Hormones/metabolism , Disease Models, AnimalABSTRACT
Sex differences in the anatomy and physiology of the respiratory system have been widely reported. These intrinsic sex differences have also been shown to modulate the pathophysiology, incidence, morbidity, and mortality of several lung diseases across the life span. In this chapter, we describe the epidemiology of sex differences in respiratory diseases including neonatal lung disease (respiratory distress syndrome, bronchopulmonary dysplasia) and pediatric and adult disease (including asthma, cystic fibrosis, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, lung cancer, lymphangioleiomyomatosis, obstructive sleep apnea, pulmonary arterial hypertension, and respiratory viral infections such as respiratory syncytial virus, influenza, and SARS-CoV-2). We also discuss the current state of research on the mechanisms underlying the observed sex differences in lung disease susceptibility and severity and the importance of considering both sex and gender variables in research studies' design and analysis.
Subject(s)
COVID-19 , Pulmonary Disease, Chronic Obstructive , Adult , Child , Female , Humans , Infant, Newborn , Lung , Male , SARS-CoV-2 , Sex CharacteristicsABSTRACT
Inflammatory lung diseases affect men and women disproportionately, suggesting that fluctuations of circulating hormone levels mediate inflammatory responses. Studies have shown that ozone exposure contributes to lung injury and impairment of innate immunity with differential effects in men and women. Here, we hypothesized that 17ß-estradiol enhances inflammation and airway hyperresponsiveness (AHR), triggered by ozone exposure, in the female lung. We performed gonadectomy and hormone treatment (17ß-estradiol, 2 wk) in C57BL/6J female and male mice and exposed animals to 1 ppm of ozone or filtered air for 3 h. Twenty-four hours later, we tested lung function, inflammatory gene expression, and changes in bronchoalveolar lavage fluid (BALF). We found increased AHR and expression of inflammatory genes after ozone exposure. These changes were higher in females and were affected by gonadectomy and 17ß-estradiol treatment in a sex-specific manner. Gonadectomized male mice displayed higher AHR and inflammatory gene expression than controls exposed to ozone; 17ß-estradiol treatment did not affect this response. In females, ovariectomy reduced ozone-induced AHR, which was restored by 17ß-estradiol treatment. Ozone exposure also increased BALF lipocalin-2, which was reduced in both male and female gonadectomized mice. Treatment with 17ß-estradiol increased lipocalin-2 levels in females but lowered them in males. Gonadectomy also reduced ozone-induced expression of lung IL-6 and macrophage inflammatory protein-3 in females, which was restored by treatment with 17ß-estradiol. Together, these results indicate that 17ß-estradiol increases ozone-induced inflammation and AHR in females but not in males. Future studies examining diseases associated with air pollution exposure should consider the patient's sex and hormonal status.
Subject(s)
Estradiol/pharmacology , Estrogens/pharmacology , Lung/physiopathology , Oxidants, Photochemical/toxicity , Ozone/toxicity , Pneumonia/pathology , Animals , Female , Lung/drug effects , Male , Mice , Mice, Inbred C57BL , Pneumonia/chemically induced , Pneumonia/metabolism , Sex FactorsABSTRACT
BACKGROUND: Influenza A viruses cause life-threatening pneumonia and lung injury in the lower respiratory tract. Application of high GM-CSF levels prior to infection has been shown to reduce morbidity and mortality from pathogenic influenza infection in mice, but the mechanisms of protection and treatment efficacy have not been established. METHODS: Mice were infected intranasally with influenza A virus (PR8 strain). Supra-physiologic levels of GM-CSF were induced in the airways using the double transgenic GM-CSF (DTGM) or littermate control mice starting on 3 days post-infection (dpi). Assessment of respiratory mechanical parameters was performed using the flexiVent rodent ventilator. RNA sequence analysis was performed on FACS-sorted airway macrophage subsets at 8 dpi. RESULTS: Supra-physiologic levels of GM-CSF conferred a survival benefit, arrested the deterioration of lung mechanics, and reduced the abundance of protein exudates in bronchoalveolar (BAL) fluid to near baseline levels. Transcriptome analysis, and subsequent validation ELISA assays, revealed that excess GM-CSF re-directs macrophages from an "M1-like" to a more "M2-like" activation state as revealed by alterations in the ratios of CXCL9 and CCL17 in BAL fluid, respectively. Ingenuity pathway analysis predicted that GM-CSF surplus during IAV infection elicits expression of anti-inflammatory mediators and moderates M1 macrophage pro-inflammatory signaling by Type II interferon (IFN-γ). CONCLUSIONS: Our data indicate that application of high levels of GM-CSF in the lung after influenza A virus infection alters pathogenic "M1-like" macrophage inflammation. These results indicate a possible therapeutic strategy for respiratory virus-associated pneumonia and acute lung injury.
Subject(s)
Cell Polarity/physiology , Granulocyte-Macrophage Colony-Stimulating Factor/biosynthesis , Influenza A virus , Macrophages/metabolism , Monocytes/metabolism , Orthomyxoviridae Infections/metabolism , Animals , Female , Gene Expression , Granulocyte-Macrophage Colony-Stimulating Factor/genetics , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Mortality/trends , Orthomyxoviridae Infections/mortality , Orthomyxoviridae Infections/prevention & controlABSTRACT
BACKGROUND: Humanistic care in medicine has shown to improve healthcare outcomes. Language barriers are a significant obstacle to humanistic care, and trained medical interpreters have demonstrated to effectively bridge the gap for the vulnerable limited English proficiency (LEP) patient population. One way in which medical schools can train more humanistic physicians and provide language access is through the implementation of programs to train bilingual medical students as medical interpreters. The purpose of this prospective study was to evaluate whether such training had an impact on bilingual medical student's interpretation skills and humanistic traits. METHODS: Between 2015 and 2017, whole-day (~ 8 h) workshops on medical interpretation were offered periodically to 80 bilingual medical students at the Penn State College of Medicine. Students completed a series of questionnaires before and after the training that assessed the program's effectiveness and its overall impact on interpretation skills and humanistic traits. Students also had the opportunity to become certified medical interpreters. RESULTS: The 80 student participants were first- to third- year medical students representing 21 languages. Following training, most students felt more confident interpreting (98%) and more empathetic towards LEP patients (87.5%). Students' scores in the multiple-choice questions about medical interpretation/role of the interpreter were also significantly improved (Chi-Square test, p < 0.05). All students who decided to take the exam were able to successfully become certified interpreters. Ninety-two percent of participants reported they would recommend the program and would be willing to serve as a future "coaches" for interpreter training workshops delivered to peer students. CONCLUSIONS: Our program was successful in increasing self-reported measures of empathy and humanism in medical students. Our data suggests that implementation of medical interpreter training programs can be a successful strategy to develop of humanism in medical students, and aid in the development of sustainable language access for LEP patients.
Subject(s)
Communication Barriers , Humanism , Students, Medical , Translating , Humans , Physician-Patient Relations , Program Development , Prospective StudiesABSTRACT
BackgroundMaternal breast milk (MBM) is enriched in microRNAs, factors that regulate protein translation throughout the human body. MBM from mothers of term and preterm infants differs in nutrient, hormone, and bioactive-factor composition, but the microRNA differences between these groups have not been compared. We hypothesized that gestational age at delivery influences microRNA in MBM, particularly microRNAs involved in immunologic and metabolic regulation.MethodsMBM from mothers of premature infants (pMBM) obtained 3-4 weeks post delivery was compared with MBM from mothers of term infants obtained at birth (tColostrum) and 3-4 weeks post delivery (tMBM). The microRNA profile in lipid and skim fractions of each sample was evaluated with high-throughput sequencing.ResultsThe expression profiles of nine microRNAs in lipid and skim pMBM differed from those in tMBM. Gene targets of these microRNAs were functionally related to elemental metabolism and lipid biosynthesis. The microRNA profile of tColostrum was also distinct from that of pMBM, but it clustered closely with tMBM. Twenty-one microRNAs correlated with gestational age demonstrated limited relationships with method of delivery, but not other maternal-infant factors.ConclusionPremature delivery results in a unique MBM microRNA profile with metabolic targets. This suggests that preterm milk may have adaptive functions for growth in premature infants.
Subject(s)
MicroRNAs/metabolism , Milk, Human/metabolism , Obstetric Labor, Premature , Adult , Female , Humans , Infant, Premature , Male , PregnancyABSTRACT
Surfactant protein A (SP-A), a molecule with roles in lung innate immunity and surfactant-related functions, is encoded by two genes in humans: SFTPA1 (SP-A1) and SFTPA2 (SP-A2). The mRNAs from these genes differ in their 5'-untranslated regions (5'-UTR) due to differential splicing. The 5'-UTR variant ACD' is exclusively found in transcripts of SP-A1, but not in those of SP-A2. Its unique exon C contains two upstream AUG codons (uAUGs) that may affect SP-A1 translation efficiency. The first uAUG (u1) is in frame with the primary start codon (p), but the second one (u2) is not. The purpose of this study was to assess the impact of uAUGs on SP-A1 expression. We employed RT-qPCR to determine the presence of exon C-containing SP-A1 transcripts in human RNA samples. We also used in vitro techniques including mutagenesis, reporter assays, and toeprinting analysis, as well as in silico analyses to determine the role of uAUGs. Exon C-containing mRNA is present in most human lung tissue samples and its expression can, under certain conditions, be regulated by factors such as dexamethasone or endotoxin. Mutating uAUGs resulted in increased luciferase activity. The mature protein size was not affected by the uAUGs, as shown by a combination of toeprint and in silico analysis for Kozak sequence, secondary structure, and signal peptide and in vitro translation in the presence of microsomes. In conclusion, alternative splicing may introduce uAUGs in SP-A1 transcripts, which in turn negatively affect SP-A1 translation, possibly affecting SP-A1/SP-A2 ratio, with potential for clinical implication.
Subject(s)
Alternative Splicing/physiology , Codon, Initiator/physiology , Lung/metabolism , Pulmonary Surfactant-Associated Protein A/biosynthesis , RNA, Messenger/biosynthesis , Alternative Splicing/drug effects , Anti-Inflammatory Agents/pharmacology , Cell Line, Tumor , Dexamethasone/pharmacology , Endotoxins/pharmacology , Exons/physiology , Female , Humans , Male , Microsomes/metabolism , Mutation , Protein Sorting Signals/physiology , Pulmonary Surfactant-Associated Protein A/genetics , RNA, Messenger/geneticsABSTRACT
Sex differences in the incidence of respiratory diseases have been reported. Women are more susceptible to inflammatory lung disease induced by air pollution and show worse adverse pulmonary health outcomes than men. However, the mechanisms underlying these differences remain unknown. In the present study, we hypothesized that sex differences in the expression of lung inflammatory mediators affect sex-specific immune responses to environmental toxicants. We focused on the effects of ground-level ozone, a major air pollutant, in the expression and regulation of lung immunity genes. We exposed adult male and female mice to 2 ppm of ozone or filtered air (control) for 3 h. We compared mRNA levels of 84 inflammatory genes in lungs harvested 4 h postexposure using a PCR array. We also evaluated changes in lung histology and bronchoalveolar lavage fluid cell counts and protein content at 24 and 72 h postexposure. Our results revealed sex differences in lung inflammation triggered by ozone exposure and in the expression of genes involved in acute phase and inflammatory responses. Major sex differences were found in the expression of neutrophil-attracting chemokines (Ccl20, Cxcl5, and Cxcl2), the proinflammatory cytokine interleukin-6, and oxidative stress-related enzymes (Ptgs2, Nos2). In addition, the phosphorylation of STAT3, known to mediate IL-6-related immune responses, was significantly higher in ozone-exposed mice. Together, our observations suggest that a differential regulation of the lung immune response could be implicated in the observed increased susceptibility to adverse health effects from ozone observed in women vs. men.
Subject(s)
Air Pollutants/toxicity , Inflammation Mediators/metabolism , Lung/metabolism , Ozone/toxicity , Pneumonia/metabolism , Animals , Capillary Permeability , Female , Interleukins/genetics , Interleukins/metabolism , Lung/immunology , Lung/pathology , Male , Mice, Inbred C57BL , Neutrophil Infiltration , Oxidative Stress , Pneumonia/chemically induced , Pneumonia/immunology , Receptors, Pattern Recognition/genetics , Receptors, Pattern Recognition/metabolism , STAT3 Transcription Factor/metabolism , Sex Characteristics , Transcription Factors/genetics , Transcription Factors/metabolism , TranscriptomeABSTRACT
Knowledge of the methylation profile of genes allow for the identification of biomarkers that may guide diagnosis and effective treatment of disease. Human surfactant protein A (SP-A) plays an important role in lung homeostasis and immunity, and is encoded by two genes (SFTPA1 and SFTPA2). The goal of this study was to identify differentially methylated CpG sites in the promoter region of the SFTPA2 gene in lung cancer tissue, and to determine the correlation between the promoter's methylation profile and gene expression. For this, we collected 28 pairs of cancerous human lung tissue and adjacent noncancerous (NC) lung tissue: 17 adenocarcinoma (AC), 9 squamous cell carcinoma (SCC), and 2 AC with SCC features, and we evaluated DNA methylation of the SFTPA2 promoter region by bisulfite conversion. Our results identified a higher methylation ratio in one CpG site of the SFTPA2 gene in cancerous tissue versus NC tissue (0.36 versus 0.11, p = 0.001). When assessing AC samples, we also found cancerous tissues associated with a higher methylation ratio (0.43 versus 0.10, p = 0.02). In the SCC group, although cancerous tissue showed a higher methylation ratio (0.22 versus 0.11), this difference was not statistically significant (p = 0.35). Expression of SFTPA2 mRNA and total SP-A protein was significantly lower in cancer tissue when compared to adjacent NC tissue (p < 0.001), and correlated with the hypermethylated status of an SFTPA2 CpG site in AC samples. The findings of this pilot study may hold promise for future use of SFTPA2 as a biomarker for the diagnosis of lung cancer.
Subject(s)
DNA Methylation/genetics , Gene Expression/genetics , Lung Neoplasms/genetics , Pulmonary Surfactant-Associated Protein A/genetics , Transcriptome/genetics , Adenocarcinoma/genetics , Base Sequence , Biomarkers, Tumor/genetics , Carcinoma, Squamous Cell/genetics , CpG Islands/genetics , Gene Expression Regulation, Neoplastic/genetics , Humans , Molecular Sequence Data , Pilot Projects , Promoter Regions, Genetic/genetics , RNA, Messenger/geneticsABSTRACT
Surfactant protein A (SP-A) plays a vital role in maintaining normal lung function and in host defense. Two genes encode SP-A in humans (SFTPA1, SFTPA2), and several gene variants have been identified for these. We have previously shown that sequence elements of SFTPA1 and SFTPA2 3' untranslated regions (UTRs) differentially affect translation efficiency in vitro. Polymorphisms at the 3'UTRs of mRNA variants may account for differential binding of miRNAs, a class of small noncoding RNAs that regulate gene expression. In this work, we generated 3'UTR reporter constructs of the SFTPA1 and SFTPA2 variants most frequently found in the population, as well as mutants of a previously described 11-nt indel element (refSNP rs368700152). Reporter constructs were transfected in NCI-H441 cells in the presence or absence of miRNA mimics, and reporter gene expression was analyzed. We found that human miRNA mir-767 negatively affected expression of constructs containing SFTPA1 and SFTPA2 variants, whereas mir-4507 affected only constructs with 3'UTRs of SFTPA1 variants 6A, 6A(3), and 6A(4) (not containing the 11-nt element). Three miRNAs (mir-183, mir-449b, and mir-612) inhibited expression of recombinants of SFTPA2 variants and the SFTPA1 variant 6A(2), all containing the 11-nt element. Similar results were obtained for SP-A expression when these miRNAs were transfected in Chinese hamster ovary cells expressing SFTPA1 or SFTPA2 variants or in NCI-H441 cells (genotype 1A(5)/1A(5)-6A(4)/6A(4)). Moreover, transfection with a specific antagomir (antagomir-183) reversed the effects of mir-183 on SP-A mRNA levels. Our results indicate that sequence variability at the 3'UTR of SP-A variants differentially affects miRNA regulation of gene expression.
Subject(s)
3' Untranslated Regions/genetics , Gene Expression Regulation/genetics , MicroRNAs/genetics , Pulmonary Surfactant-Associated Protein A/genetics , Animals , Base Sequence , CHO Cells , Cell Line, Tumor , Cricetinae , Cricetulus , Humans , Molecular Sequence Data , Polymorphism, Single Nucleotide , Protein Biosynthesis/genetics , RNA, Messenger/genetics , Sequence AlignmentABSTRACT
Human surfactant protein A (SP-A) plays an important role in surfactant metabolism and lung innate immunity. SP-A is synthesized and secreted by alveolar type II (ATII) cells, one of the two cell types of the distal lung epithelium (ATII and ATI). We have shown that miRNA interactions with sequence polymorphisms on the SP-A mRNA 3'UTRs mediate differential expression of SP-A1 and SP-A2 gene variants in vitro. In the present study, we describe a physiologically relevant model to study miRNA regulation of SP-A in human ATII. For these studies, we purified and cultured human ATII on an air-liquid interface matrix (A/L) or plastic wells without matrix (P). Gene expression analyses confirmed that cells cultured in A/L maintained the ATII phenotype for over 5 days, whereas P-cultured cells differentiated to ATI. When we transfected ATII with siRNAs to inhibit the expression of Drosha, a critical effector of miRNA maturation, the levels of SP-A mRNA and protein increased in a time dependent manner. We next characterized cultured ATII and ATI by studying expression of 1,066 human miRNAs using miRNA PCR arrays. We detected expression of >300 miRNAs with 24 miRNAs differentially expressed in ATII versus ATI, 12 of which predicted to bind SP-A 3'UTRs, indicating that these may be implicated in SP-A downregulation in ATI. Thus, miRNAs not only affect SP-A expression, but also may contribute to the maintenance of the ATII cell phenotype and/or the trans-differentiation of ATII to ATI cells, and may represent new molecular markers that distinguish ATII and ATI.
Subject(s)
Pulmonary Alveoli/metabolism , Pulmonary Surfactant-Associated Protein A/genetics , Pulmonary Surfactant-Associated Protein A/metabolism , Ribonuclease III/genetics , Ribonuclease III/metabolism , 3' Untranslated Regions/genetics , Cell Differentiation/genetics , Cells, Cultured , Epithelial Cells/metabolism , Epithelium/metabolism , Humans , MicroRNAs/genetics , Phenotype , Pilot Projects , RNA, Messenger/genetics , RNA, Small Interfering/geneticsABSTRACT
Lung cancer remains a global public health concern with significant research focus on developing better diagnosis/prognosis biomarkers and therapeutical targets. Circular RNAs (circRNAs) are a type of single-stranded RNA molecules that covalently closed and have ubiquitous expression. These molecules have been implicated in a variety of disease mechanisms, including lung cancer, as they exhibit oncogenic or tumor suppressor characteristics. Recent research has shown an important role that circRNAs play at different stages of lung cancer, particularly in lung adenocarcinoma. In this review, we summarize the latest research on circRNAs and their roles within lung cancer diagnosis, as well as on disease mechanisms. We also discuss the knowledge gaps on these topics and possible future research directions.
ABSTRACT
Cardiovascular diseases (CVDs) are the leading cause of death worldwide. CVDs are promoted by the accumulation of lipids and immune cells in the endothelial space resulting in endothelial dysfunction. Endothelial cells are important components of the vascular endothelium, that regulate the vascular flow. The imbalance in the production of vasoactive substances results in the loss of vascular homeostasis, leading the endothelial dysfunction. Thus, endothelial dysfunction plays an essential role in the development of atherosclerosis and can be triggered by different cardiovascular risk factors. On the other hand, the 17ß-estradiol (E2) hormone has been related to the regulation of vascular tone through different mechanisms. Several compounds can elicit estrogenic actions similar to those of E2. For these reasons, they have been called endocrine-disrupting compounds (EDCs). This review aims to provide up-to-date information about how different EDCs affect endothelial function and their mechanistic roles in the context of CVDs.
Subject(s)
Cardiovascular Diseases , Endocrine Disruptors , Phthalic Acids , Humans , Parabens/toxicity , Endothelial Cells , Estradiol , Cardiovascular Diseases/chemically induced , Endothelium, Vascular/physiology , Endocrine Disruptors/toxicityABSTRACT
The emerging concern about chemicals in electronic cigarettes, even those without nicotine, demands the development of advanced criteria for their exposure and risk assessment. This study aims to highlight the sensitivity of lung nuclear receptors (NRs) to electronic cigarette e-liquids, independent of nicotine presence, and the influence of the sex variable on these effects. Adult male and female C57BL/6J mice were exposed to electronic cigarettes with 0%, 3%, and 6% nicotine daily (70 mL, 3.3 s, 1 puff per min/30 min) for 14 days, using the inExpose full body chamber (SCIREQ). Following exposure, lung tissues were harvested, and RNA extracted. The expression of 84 NRs was determined using the RT2 profiler mRNA array (Qiagen). Results exhibit a high sensitivity to e-liquid exposure irrespective of the presence of nicotine, with differential expression of NRs, including one (females) and twenty-four (males) in 0% nicotine groups compared to non-exposed control mice. However, nicotine-dependent results were also significant with seven NRs (females), fifty-three NRs (males) in 3% and twenty-three NRs (female) twenty-nine NRs (male) in 6% nicotine groups, compared to 0% nicotine mice. Sex-specific changes were significant, but sex-related differences were not observed. The study provides a strong rationale for further investigation.
Subject(s)
Aerosols , Electronic Nicotine Delivery Systems , Lung , Mice, Inbred C57BL , Nicotine , Receptors, Cytoplasmic and Nuclear , Animals , Female , Male , Pilot Projects , Mice , Lung/metabolism , Lung/drug effects , Receptors, Cytoplasmic and Nuclear/metabolism , Receptors, Cytoplasmic and Nuclear/genetics , Sex FactorsABSTRACT
Human surfactant protein A, an innate immunity molecule, is encoded by two genes: SFTPA1 (SP-A1) and SFTPA2 (SP-A2). The 5' untranslated (5'UTR) splice variant of SP-A2 (ABD), but not of SP-A1 (AD), contains exon B (eB), which is an enhancer for transcription and translation. We investigated whether eB contains cis-regulatory elements that bind trans-acting factors in a sequence-specific manner as well as the role of the eB mRNA secondary structure. Binding of cytoplasmic NCI-H441 proteins to wild-type eB, eB mutant, AD, and ABD 5'UTR mRNAs were studied by RNA electromobility shift assays (REMSAs). The bound proteins were identified by mass spectroscopy and specific antibodies (Abs). We found that 1) proteins bind eB mRNA in a sequence-specific manner, with two cis-elements identified within eB to be important; 2) eB secondary structure is necessary for binding; 3) mass spectroscopy and specific Abs in REMSAs identified 14-3-3 proteins to bind (directly or indirectly) eB and the natural SP-A2 (ABD) splice variant but not the SP-A1 (AD) splice variant; 4) other ribosomal and cytoskeletal proteins, and translation factors, are also present in the eB mRNA-protein complex; 5) knockdown of 14-3-3 ß/α isoform resulted in a downregulation of SP-A2 expression. In conclusion, proteins including the 14-3-3 family bind two cis-elements within eB of hSP-A2 mRNA in a sequence- and secondary structure-specific manner. Differential regulation of SP-A1 and SP-A2 is mediated by the 14-3-3 protein family as well as by a number of other proteins that bind UTRs with or without eB mRNA.
Subject(s)
14-3-3 Proteins/metabolism , Exons/genetics , Pulmonary Surfactant-Associated Protein A/genetics , RNA, Messenger/metabolism , 5' Untranslated Regions/physiology , Base Sequence , Cell Line, Tumor , Humans , Protein Structure, SecondaryABSTRACT
BACKGROUND: Electronic cigarettes (e-cigarettes) comprise a variety of products designed to deliver nicotine, flavorings, and other substances. To date, multiple epidemiological and experimental studies have reported a variety of health issues associated with their use, including respiratory toxicity, exacerbation of respiratory conditions, and behavioral and physiological effects. While some of these effects appear to be sex- and/or gender-related, only a portion of the research has been conducted considering these variables. In this review, we sought to summarize the available literature on sex-specific effects and sex and gender differences, including predictors and risk factors, effects on organ systems, and behavioral effects. METHODS: We searched and selected articles from 2018-2023 that included sex as a variable or reported sex differences on e-cigarette-associated effects. RESULTS: We found 115 relevant studies published since 2018 that reported sex differences in a variety of outcomes. The main differences reported were related to reasons for initiation, including smoking history, types of devices and flavoring, polysubstance use, physiological responses to nicotine and toxicants in e-liquids, exacerbation of lung disease, and behavioral factors such as anxiety, depression, sexuality, and bullying. CONCLUSIONS: The available literature supports the notion that both sex and gender influence the susceptibility to the negative effects of e-cigarette use. Future research needs to consider sex and gender variables when addressing e-cigarette toxicity and other health-related consequences.
Subject(s)
Electronic Nicotine Delivery Systems , Tobacco Products , Vaping , Humans , Male , Female , Vaping/adverse effects , Vaping/epidemiology , Nicotine/adverse effects , Sex Factors , Sex Characteristics , Flavoring Agents/toxicityABSTRACT
OBJECTIVES: High-dose oral iron supplementation for patients who develop iron deficiency after bariatric surgery may induce oxidative stress in the gastrointestine. The study's objective was to test this hypothesis by determining the impact of high-dose oral iron on systemic oxidative stress. METHODS: We used archived plasma samples from a randomized controlled clinical trial (NCT02404012) comparing FeSO4 (195 mg/day, NatureMade®, West Hills, CA) with a heme iron polypeptide (HIP, 60.4 mg/day, Proferrin®, Colorado Biolabs, Lafayette, CO) for 8 weeks. Systemic oxidative stress was measured using malondialdehyde and total antioxidant capacity (MDA, Abcam, ab238537 and TAC, Abcam, ab65329 Cambridge, UK) assays. Data was log-transformed and presented as means and standard deviations; a mixed model was used to determine the effects of time (0, 2, 4, and 8 weeks) and treatment (FeSO4 versus HIP) on oxidative stress. RESULTS: The FeSO4 (N = 8) and HIP (N = 5) participants were balanced in body mass index (35.0 ± 5.5 kg/m2), race (93 % White), time post-surgery (7.3 ± 3.3 years), as well as serum concentrations of iron (P > 0.05). The FeSO4 group tended to be older (44.3 ± 4.5 years) and they had lower concentrations of serum ferritin (6.5 ± 2.7 µg/mL) than the HIP (38.2 ± 9.3 years, and 12.9 ± 16.8 µg/mL) group (P = 0.080, and P = 0.017 respectively). We observed a larger increase in serum iron in the FeSO4 group during the 8 weeks of Fe supplementation, compared to that in the HIP group (p = 0.004). We observed a decreasing trend in MDA over the 8 weeks (p = 0.080) in the FeSO4 treatment group. There were no significant differences in TAC between and within FeSO4 and HIP groups over the 8 week supplementation period. CONCLUSIONS: This preliminary study suggests that high-dose oral iron supplementation for iron deficiency does not adversely impact systemic oxidative stress in patients undergoing bariatric surgery.