Abstract 14803: Activin A Directly Impaired Human IPSC-cardiomyocyte Contractility and Electrophysiology

Activin A is a homo-dimeric TGF-β family member involved in embryonic development, tissue morphogenesis, and cellular differentiation. Levels of activin A correlate with NYHA functional classification and age dependent cardiac dysfunction; however, no data are available to demonstrate a direct role for activin A in regulating human cardiomyocyte dysfunction. The purpose of this study was to characterize the functional impact of chronic activin A on human cardiomyocyte electrophysiology, contractility, calcium handling, and gene expression. Methods: Human iPSC-cardiomyocytes (iPSC-CM) were supplied by Fujifilm Cellular Dynamics. Calcium transients were captured using the FLIPR Tetra imager and impedance/electrophysiology by the Nanion CardioExcyte96. Cells were plated (50k/well), allowed to reach a stable synchronous monolayer, and dosed with 1nM of activin A (A) at each media change alongside a media control (C) or in the presence of inhibitory anti-activin A antibodies. Gene expression was determined using Taqman. Results: Activin A reduced contractile impedance amplitude (C=9.82±0.30, A=6.82±0.21 Ohms, p<0.01) and slowed relaxation velocity kinetics (C=48.78±2.65, A=35.07±1.07 Ohms/sec, p<0.01). Elongated extracellular field potential durations were observed with activin A treatment compared to control (C=0.49±0.02, A=0.56±0.01 sec, p<0.01). Impaired calcium handling peak amplitude (C=609±99, A= C=447±33 RFU, p<0.05) was also observed alongside slower calcium flux falling times (C=0.52±0.05, A=0.70±0.04 sec, p<0.01). An inhibitory activin-A antibody restored all parameters to control values. Chronic exposure of iPSC-CM to activin A reduced expression of calcium handling genes RYR2 (C=1.01±0.07, A=0.75±0.03, p<0.01) and ATP2A2 (C=1.00±0.05, A=0.89±0.05, p<0.05). Conclusions: Chronic activin A treatment reduced cardiomyocyte contractile amplitude, slowed contractile kinetics, impaired cardiomyocyte calcium handling, elongated the action potential, and reduced expression of genes regulating key calcium handling proteins. These data demonstrate that elevated levels of activin A can directly act on cardiomyocytes and therefore may contribute to cardiac dysfunction in heart failure and aged populations.

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Fecal transplantation and butyrate improve neuropathic pain, modify immune cell profile, and gene expression in the PNS of obese mice

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2006065117 ◽

2020 ◽

Vol 117 (42) ◽

pp. 26482-26493 ◽

Cited By ~ 1

Author(s):

Raiza R. Bonomo ◽

Tyler M. Cook ◽

Chaitanya K. Gavini ◽

Chelsea R. White ◽

Jacob R. Jones ◽

...

Keyword(s):

Gene Expression ◽

Gut Microbiome ◽

Molecular Mechanisms ◽

Immune Cell ◽

Underlying Disease ◽

Calcium Handling ◽

Fecal Transplantation ◽

Fecal Microbiome ◽

Expression Of Genes ◽

Induced Changes

Obesity affects over 2 billion people worldwide and is accompanied by peripheral neuropathy (PN) and an associated poorer quality of life. Despite high prevalence, the molecular mechanisms underlying the painful manifestations of PN are poorly understood, and therapies are restricted to use of painkillers or other drugs that do not address the underlying disease. Studies have demonstrated that the gut microbiome is linked to metabolic health and its alteration is associated with many diseases, including obesity. Pathologic changes to the gut microbiome have recently been linked to somatosensory pain, but any relationships between gut microbiome and PN in obesity have yet to be explored. Our data show that mice fed a Western diet developed indices of PN that were attenuated by concurrent fecal microbiome transplantation (FMT). In addition, we observed changes in expression of genes involved in lipid metabolism and calcium handling in cells of the peripheral nerve system (PNS). FMT also induced changes in the immune cell populations of the PNS. There was a correlation between an increase in the circulating short-chain fatty acid butyrate and pain improvement following FMT. Additionally, butyrate modulated gene expression and immune cells in the PNS. Circulating butyrate was also negatively correlated with distal pain in 29 participants with varied body mass index. Our data suggest that the metabolite butyrate, secreted by the gut microbiome, underlies some of the effects of FMT. Targeting the gut microbiome, butyrate, and its consequences may represent novel viable approaches to prevent or relieve obesity-associated neuropathies.

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Different Impacts of MucR Binding to the babR and virB Promoters on Gene Expression in Brucella abortus 2308

Biomolecules ◽

10.3390/biom10050788 ◽

2020 ◽

Vol 10 (5) ◽

pp. 788 ◽

Cited By ~ 1

Author(s):

Giorgia Borriello ◽

Veronica Russo ◽

Rubina Paradiso ◽

Marita Georgia Riccardi ◽

Daniela Criscuolo ◽

...

Keyword(s):

Gene Expression ◽

Brucella Abortus ◽

Virulence Genes ◽

Gene Promoter ◽

Direct Interaction ◽

Mobility Shift ◽

Direct Role ◽

Expression Of Genes ◽

Electrophoretic Mobility Shift Assays

The protein MucR from Brucella abortus has been described as a transcriptional regulator of many virulence genes. It is a member of the Ros/MucR family comprising proteins that control the expression of genes important for the successful interaction of α-proteobacteria with their eukaryotic hosts. Despite clear evidence of the role of MucR in repressing virulence genes, no study has been carried out so far demonstrating the direct interaction of this protein with the promoter of its target gene babR encoding a LuxR-like regulator repressing virB genes. In this study, we show for the first time the ability of MucR to bind the promoter of babR in electrophoretic mobility shift assays demonstrating a direct role of MucR in repressing this gene. Furthermore, we demonstrate that MucR can bind the virB gene promoter. Analyses by RT-qPCR showed no significant differences in the expression level of virB genes in Brucella abortus CC092 lacking MucR compared to the wild-type Brucella abortus strain, indicating that MucR binding to the virB promoter has little impact on virB gene expression in B. abortus 2308. The MucR modality to bind the two promoters analyzed supports our previous hypothesis that this is a histone-like protein never found before in Brucella.

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Ouabain-Induced Gene Expression Changes in Human iPSC-Derived Neuron Culture Expressing Dopamine and cAMP-Regulated Phosphoprotein 32 and GABA Receptors

Brain Sciences ◽

10.3390/brainsci11020203 ◽

2021 ◽

Vol 11 (2) ◽

pp. 203

Author(s):

Alexander V. Lopachev ◽

Maria A. Lagarkova ◽

Olga S. Lebedeva ◽

Margarita A. Ezhova ◽

Rogneda B. Kazanskaya ◽

...

Keyword(s):

Gene Expression ◽

Gaba Receptors ◽

Neurite Growth ◽

Signaling Cascades ◽

Neuronal Maturation ◽

Medium Spiny Neurons ◽

Neuron Culture ◽

Spiny Neurons ◽

Expression Of Genes ◽

Human Ipsc

Cardiotonic steroids (CTS) are specific inhibitors and endogenous ligands of a key enzyme in the CNS—the Na+, K+-ATPase, which maintains and creates an ion gradient on the plasma membrane of neurons. CTS cause the activation of various signaling cascades and changes in gene expression in neurons and other cell types. It is known that intracerebroventricular injection of cardiotonic steroid ouabain causes mania-like behavior in rodents, in part due to activation of dopamine-related signaling cascades in the dopamine and cAMP-regulated phosphoprotein 32 (DARPP-32) expressing medium spiny neurons in the striatum. Dopaminergic projections in the striatum innervate these GABAergic medium spiny neurons. The objective of this study was to assess changes in the expression of all genes in human iPSC-derived expressing DARPP-32 and GABA receptors neurons under the influence of ouabain. We noted a large number of statistically significant upregulated and downregulated genes after a 16-h incubation with non-toxic concentration (30 nM) of ouabain. These changes in the transcriptional activity were accomplished with activation of MAP-kinase ERK1/2 and transcriptional factor cAMP response element-binding protein (CREB). Thus, it can be concluded that 30 nM ouabain incubated for 16 h with human iPSC-derived expressing DARPP-32 and GABA receptors neurons activates genes associated with neuronal maturation and synapse formation, by increasing the expression of genes associated with translation, vesicular transport, and increased electron transport chain function. At the same time, the expression of genes associated with proliferation, migration, and early development of neurons decreases. These data indicate that non-toxic concentrations of ouabain may induce neuronal maturation, neurite growth, and increased synaptogenesis in dopamine-receptive GABAergic neurons, suggesting formation of plasticity and the establishment of new neuronal junctions.

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An Efficient Method for the Differentiation of Human iPSC-Derived Endoderm toward Enterocytes and Hepatocytes

Cells ◽

10.3390/cells10040812 ◽

2021 ◽

Vol 10 (4) ◽

pp. 812

Author(s):

Shimeng Qiu ◽

Yaling Li ◽

Yuki Imakura ◽

Shinji Mima ◽

Tadahiro Hashita ◽

...

Keyword(s):

Small Intestine ◽

Activin A ◽

Double Positive ◽

Differentiation Method ◽

Drug Metabolizing Enzyme ◽

Human Ipscs ◽

Novel Method ◽

Induced Pluripotent ◽

Metabolizing Enzyme ◽

Human Ipsc

The endoderm, differentiated from human induced pluripotent stem cells (iPSCs), can differentiate into the small intestine and liver, which are vital for drug absorption and metabolism. The development of human iPSC-derived enterocytes (HiEnts) and hepatocytes (HiHeps) has been reported. However, pharmacokinetic function-deficiency of these cells remains to be elucidated. Here, we aimed to develop an efficient differentiation method to induce endoderm formation from human iPSCs. Cells treated with activin A for 168 h expressed higher levels of endodermal genes than those treated for 72 h. Using activin A (days 0–7), CHIR99021 and PI−103 (days 0–2), and FGF2 (days 3–7), the hiPSC-derived endoderm (HiEnd) showed 97.97% CD−117 and CD−184 double-positive cells. Moreover, HiEnts derived from the human iPSC line Windy had similar or higher expression of small intestine-specific genes than adult human small intestine. Activities of the drug transporter P-glycoprotein and drug-metabolizing enzyme cytochrome P450 (CYP) 3A4/5 were confirmed. Additionally, Windy-derived HiHeps expressed higher levels of hepatocyte- and pharmacokinetics-related genes and proteins and showed higher CYP3A4/5 activity than those derived through the conventional differentiation method. Thus, using this novel method, the differentiated HiEnts and HiHeps with pharmacokinetic functions could be used for drug development.

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Impact of Environmental Stressors on Gene Expression in the Embryo of the Italian Wall Lizard

Applied Sciences ◽

10.3390/app11114723 ◽

2021 ◽

Vol 11 (11) ◽

pp. 4723

Author(s):

Rosaria Scudiero ◽

Chiara Maria Motta ◽

Palma Simoniello

Keyword(s):

Gene Expression ◽

Protein Interactions ◽

Membrane Trafficking ◽

Translational Regulation ◽

Cellular Protection ◽

Terrestrial Vertebrate ◽

Expression Of Genes ◽

Stress Changes ◽

Probable Consequence ◽

The Impact

The cleidoic eggs of oviparous reptiles are protected from the external environment by membranes and a parchment shell permeable to water and dissolved molecules. As a consequence, not only physical but also chemical insults can reach the developing embryos, interfering with gene expression. This review provides information on the impact of the exposure to cadmium contamination or thermal stress on gene expression during the development of Italian wall lizards of the genus Podarcis. The results obtained by transcriptomic analysis, although not exhaustive, allowed to identify some stress-reactive genes and, consequently, the molecular pathways in which these genes are involved. Cadmium-responsive genes encode proteins involved in cellular protection, metabolism and proliferation, membrane trafficking, protein interactions, neuronal transmission and plasticity, immune response, and transcription regulatory factors. Cold stress changes the expression of genes involved in transcriptional/translational regulation and chromatin remodeling and inhibits the transcription of a histone methyltransferase with the probable consequence of modifying the epigenetic control of DNA. These findings provide transcriptome-level evidence of how terrestrial vertebrate embryos cope with stress, giving a key to use in population survival and environmental change studies. A better understanding of the genes contributing to stress tolerance in vertebrates would facilitate methodologies and applications aimed at improving resistance to unfavourable environments.

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Transversal gene expression panel to evaluate intestinal health in broiler chickens in different challenging conditions

Scientific Reports ◽

10.1038/s41598-021-85872-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

L. Criado-Mesas ◽

N. Abdelli ◽

A. Noce ◽

M. Farré ◽

J. F. Pérez ◽

...

Keyword(s):

Gene Expression ◽

Immune Response ◽

Barrier Function ◽

Nutrient Transport ◽

Gene Expression Pattern ◽

Vaccine Dose ◽

Special Focus ◽

Intestinal Health ◽

Barrier Failure ◽

Expression Of Genes

AbstractThere is a high interest on gut health in poultry with special focus on consequences of the intestinal diseases, such as coccidiosis and C. perfringens-induced necrotic enteritis (NE). We developed a custom gene expression panel, which could provide a snapshot of gene expression variation under challenging conditions. Ileum gene expression studies were performed through high throughput reverse transcription quantitative real-time polymerase chain reaction. A deep review on the bibliography was done and genes related to intestinal health were selected for barrier function, immune response, oxidation, digestive hormones, nutrient transport, and metabolism. The panel was firstly tested by using a nutritional/Clostridium perfringens model of intestinal barrier failure (induced using commercial reused litter and wheat-based diets without exogenous supplementation of enzymes) and the consistency of results was evaluated by another experiment under a coccidiosis challenge (orally gavaged with a commercial coccidiosis vaccine, 90× vaccine dose). Growth traits and intestinal morphological analysis were performed to check the gut barrier failure occurrence. Results of ileum gene expression showed a higher expression in genes involved in barrier function and nutrient transport in chickens raised in healthy conditions, while genes involved in immune response presented higher expression in C.perfringens-challenged birds. On the other hand, the Eimeria challenge also altered the expression of genes related to barrier function and metabolism, and increased the expression of genes related to immune response and oxidative stress. The panel developed in the current study gives us an overview of genes and pathways involved in broiler response to pathogen challenge. It also allows us to deep into the study of differences in gene expression pattern and magnitude of responses under either a coccidial vaccine or a NE.

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Selection of reference genes for gene expression analysis in Liriodendron hybrids’ somatic embryogenesis and germinative tissues

Scientific Reports ◽

10.1038/s41598-021-84518-w ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Tingting Li ◽

Weigao Yuan ◽

Shuai Qiu ◽

Jisen Shi

Keyword(s):

Gene Expression ◽

Somatic Embryogenesis ◽

Reference Genes ◽

Gene Selection ◽

Elongation Factor ◽

Reverse Transcription Pcr ◽

Expression Of Genes ◽

Suitable Reference ◽

Functional Analyses ◽

Elongation Factor 1

AbstractThe differential expression of genes is crucial for plant somatic embryogenesis (SE), and the accurate quantification of gene expression levels relies on choosing appropriate reference genes. To select the most suitable reference genes for SE studies, 10 commonly used reference genes were examined in synchronized somatic embryogenic and subsequent germinative cultures of Liriodendron hybrids by using quantitative real-time reverse transcription PCR. Four popular normalization algorithms: geNorm, NormFinder, Bestkeeper and Delta-Ct were used to select and validate the suitable reference genes. The results showed that elongation factor 1-gamma, histone H1 linker protein, glyceraldehyde-3-phosphate dehydrogenase and α-tubulin were suitable for SE tissues, while elongation factor 1-gamma and actin were best for the germinative organ tissues. Our work will benefit future studies of gene expression and functional analyses of SE in Liriodendron hybrids. It is also serves as a guide of reference gene selection in early embryonic gene expression analyses for other woody plant species.

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Controllability of Boolean networks with intermittent disturbances

Modern Physics Letters B ◽

10.1142/s0217984920503091 ◽

2020 ◽

Vol 34 (28) ◽

pp. 2050309

Author(s):

Tao You ◽

Hailun Zhang ◽

Mingyu Yang ◽

Xiao Wang ◽

Yangming Guo

Keyword(s):

Gene Expression ◽

Boolean Network ◽

Boolean Networks ◽

Genetic Mutations ◽

Intermittent Control ◽

Mathematical Tool ◽

Pulse Control ◽

Expression Of Genes ◽

Complicated Process ◽

Living Being

In biological systems, gene expression is an important subject. In order to clarify the specific process of gene expression, mathematical tools are needed to simulate the process. The Boolean network (BN) is a good mathematical tool. In this paper, we study a Boolean network with intermittent perturbations. This is of great significance for studying genetic mutations in bioengineering. The expression of genes in the internal system of a living being is a very complicated process, and it is clear that the process is trans-ageal for humans. Through the intermittent control and pulse control of the BN, we can obtain the trajectory of gene expression better and faster, which will provide a very important theoretical basis for our next research.

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Integrating metabolomics and targeted gene expression to uncover potential biomarkers of fungal/oomycetes-associated disease susceptibility in grapevine

Scientific Reports ◽

10.1038/s41598-020-72781-2 ◽

2020 ◽

Vol 10 (1) ◽

Cited By ~ 1

Author(s):

Marisa Maia ◽

António E. N. Ferreira ◽

Rui Nascimento ◽

Filipa Monteiro ◽

Francisco Traquete ◽

...

Keyword(s):

Gene Expression ◽

Plant Defence ◽

Breeding Programs ◽

Analysis Group ◽

Expression Of Genes ◽

Leucoanthocyanidin Reductase ◽

Defence Responses ◽

Targeted Gene Expression ◽

Plant Defence Responses ◽

First Time

Abstract Vitis vinifera, one of the most cultivated fruit crops, is susceptible to several diseases particularly caused by fungus and oomycete pathogens. In contrast, other Vitis species (American, Asian) display different degrees of tolerance/resistance to these pathogens, being widely used in breeding programs to introgress resistance traits in elite V. vinifera cultivars. Secondary metabolites are important players in plant defence responses. Therefore, the characterization of the metabolic profiles associated with disease resistance and susceptibility traits in grapevine is a promising approach to identify trait-related biomarkers. In this work, the leaf metabolic composition of eleven Vitis genotypes was analysed using an untargeted metabolomics approach. A total of 190 putative metabolites were found to discriminate resistant/partial resistant from susceptible genotypes. The biological relevance of discriminative compounds was assessed by pathway analysis. Several compounds were selected as promising biomarkers and the expression of genes coding for enzymes associated with their metabolic pathways was analysed. Reference genes for these grapevine genotypes were established for normalisation of candidate gene expression. The leucoanthocyanidin reductase 2 gene (LAR2) presented a significant increase of expression in susceptible genotypes, in accordance with catechin accumulation in this analysis group. Up to our knowledge this is the first time that metabolic constitutive biomarkers are proposed, opening new insights into plant selection on breeding programs.

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Distinct gene expression patterns in chronic lymphocytic leukemia defined by usage of specific VH genes

Blood ◽

10.1182/blood-2005-04-1483 ◽

2006 ◽

Vol 107 (5) ◽

pp. 2090-2093 ◽

Cited By ~ 39

Author(s):

Dirk Kienle ◽

Axel Benner ◽

Alexander Kröber ◽

Dirk Winkler ◽

Daniel Mertens ◽

...

Keyword(s):

Gene Expression ◽

Chronic Lymphocytic Leukemia ◽

Expression Patterns ◽

Gene Expression Pattern ◽

Cell Receptor ◽

Lymphocytic Leukemia ◽

Expression Of Genes ◽

Vh Genes ◽

Signaling Activation

The mutation status and usage of specific VH genes such as V3-21 and V1-69 are potentially independent pathogenic and prognostic factors in chronic lymphocytic leukemia (CLL). To investigate the role of antigenic stimulation, we analyzed the expression of genes involved in B-cell receptor (BCR) signaling/activation, cell cycle, and apoptosis control in CLL using these specific VH genes compared to VH mutated (VH-MUT) and VH unmutated (VH-UM) CLL not using these VH genes. V3-21 cases showed characteristic expression differences compared to VH-MUT (up: ZAP70 [or ZAP-70]; down: CCND2, P27) and VH-UM (down: PI3K, CCND2, P27, CDK4, BAX) involving several BCR-related genes. Similarly, there was a marked difference between VH unmutated cases using the V1-69 gene and VH-UM (up: FOS; down: BLNK, SYK, CDK4, TP53). Therefore, usage of specific VH genes appears to have a strong influence on the gene expression pattern pointing to antigen recognition and ongoing BCR stimulation as a pathogenic factor in these CLL subgroups.

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