Abstract 13453: Estrogen Regulates Expression of Fibrotic Markers in Ips-derived Cardiac Fibroblasts and Cardiomyocytes

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
Mariana A Argenziano ◽  
Mariana Burgos Angulo ◽  
jiajia yang ◽  
Alexander Bertalovitz ◽  
Thomas McDonald
Keyword(s):  
Gene Expression ◽  
Wound Healing ◽  
Cardiac Fibroblasts ◽  
Epidemiologic Studies ◽  
Functional Studies ◽  
Fibrillar Collagens ◽  
Menopausal Women ◽  
Before And After ◽  
Induced Pluripotent ◽  
Beating Frequency

Introduction: Cardiovascular disease (CVD) is the leading cause of death among women. Epidemiologic studies indicate that pre-menopausal women are protected against the development of CVD when compared to age-matched men. Women show less cardiac remodeling, with downregulation in fibrosis and inflammation pathways. Remodeling is manifested by morphology and electrophysiological changes including hypertrophy, apoptosis, fibroblast proliferation, fibrosis and accumulation of fibrillar collagens. Hypothesis: We hypothesize that female protection against CVD is associated with estrogen levels as the incidence and severity of CVD increases after menopause. Methods: Induced pluripotent stem cells (IPSC) from male (M) and female (F) controls were differentiated to cardiomyocytes (IPSC-CMs) and cardiac fibroblasts (IPSC-CFs). Samples were cultured for 30 days and collected for analysis before and after 48 hs incubation with 1uM of 17-b-estradiol (EST). In IPSC-CMs we analyzed gene expression, morphology and beating frequency. Gene expression, morphology and wound healing were analyzed in IPSC-CFs. Results: Expression of ESR1 , and MYH7 was higher in female IPSC-CMs with upregulation after EST treatment in females but not males. Male IPSC-CMs showed higher expression of COL1A1 , CX43 , MYH6 and RYR2 ; MYH6 , RYR2 and COL1A1 were downregulated after EST treatment. No differences were observed in morphology or beating frequency before or after EST treatment (F:27.5±4.5 bpm vs M:22±2.5 bpm). In IPSC-CFs, males showed higher expression of the fibrotic markers COL1A1 , POSTN, SMAD2 and CCL2 ; EST treatment decreased the expression of POSTN and SMAD2 . Wound healing assays showed a decrease of the wound healing % in women after EST treatment (CT F:86.4±19.7% vs EST F:57.43±10.1%, at 8 hs, *p<0.05). Male wound healing rate was slower than women in control and treated groups, with no changes after EST treatment (CT F: 86.4±19.7% vs CT M: 51.65±18.4%, at 8 hs, *p<0.05). Conclusions: Estrogen treatment exerts an effect on gene expression in female and male IPSC-CMs and IPSC-CFs with men showing higher expression of fibrotic markers, which are down-regulated by EST. Further functional studies will shed light on the phenotypic effects of gene expression changes.

Identification of Genes During the Reprogramming of Fibroblast using Bioinformatics Analysis

2021 ◽  
Author(s):  
Peng Wu ◽  
Xiyalatu Sai ◽  
Xiaoying Tang ◽  
Huiming Guo ◽  
Huanlei Huang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Profiles ◽  
Human Fibroblasts ◽  
Gene Expression Profiles ◽  
Gene Expression Omnibus ◽  
Reprogramming Efficiency ◽  
Before And After ◽  
Induced Pluripotent ◽  
Microarray Datasets ◽  
Core Genes

Abstract Bioinformatics method was used to screen Human induced pluripotent stem cells reprogrammed by Human Fibroblasts (FBs). The reprogramming efficiency was explored by difference expressed genes (DEGs) before and after the expression of hiPSCs. Likewise, microarray datasets GSE34309, GSE43996, and GSE56805 datasets were downloaded from Gene Expression Omnibus (GEO) and GEO2R to screen the differentially expressed DEGs. Further, Gene Ontology (GO) and KEGG (Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways were analyzed by David database. Likewise, to explore the correlation between DEGs, and the STRING database software was used to construct the protein-protein internet (PPI), and the Cytoscape_v3.7.2 Cytohubba plug-in was used to screen out the core genes. A total of 622 DEGs were identified, including 344 up regulated genes and 278 down-regulated genes. DEGs were mainly concentrated in the cell cycle and PI3K-Akt signaling pathway. Among the ten core genes screened, CDK1, IL6, EGFR, FN1, CDH1, SOX2, BRCA1, EZH2, CD44, and CCNB1 were most significant in the reprogramming of FBs into iPSCs. Regardless underline the differential of those gene expression profiles and regulatory network during FBs reprogramming could provide a theoretical basis for efficient reprogramming.

Abstract 13599: A Truncated Mutation in Filamin C Reveals Non-myocyte Contributions to Hypertrophic Cardiomyopathy in Ipsc-derived Cardiac Fibroblasts

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
Mariana A Argenziano ◽  
jiajia yang ◽  
Mariana Burgos Angulo ◽  
Alexander Bertalovitz ◽  
Thomas V McDonald
Keyword(s):  
Gene Expression ◽  
Wound Healing ◽  
Cell Migration ◽  
Hypertrophic Cardiomyopathy ◽  
Sendai Virus ◽  
Cardiac Fibroblasts ◽  
Genetic Disorder ◽  
Cell Types ◽  
Pathogenic Mutation ◽  
Human Model

Introduction: Hypertrophic cardiomyopathy (HCM) is a cardiac genetic disorder that affects 1:500 individuals and represents one of the main causes of sudden cardiac death in young adults. Several mutations have been associated with HCM, including MYH7 and FLNC . The discovery of induced pluripotent stem cells (IPSC), has provided a unique patient- and disease-specific model to study cardiac disease. Although iPSC derived cardiomyocytes (IPSC-CMs) can provide a genetically diverse human model to study mechanisms of disease, the heart relies in a three-dimensional network of cells that actively participate in the development of hypertrophy. Hypothesis: We hypothesize that mutations causing HCM affect not only CMs, but other non-myocyte cell types such as cardiac fibroblasts contributing to the phenotype of the disease. Methods: IPSCs were reprogrammed from an HCM patient harboring a pathogenic mutation on FLNC (p. Ile616Aspfs*4) using Sendai virus. Cells were differentiated to cardiomyocytes (IPSC-CMs) and cardiac fibroblasts (IPSC-CFs) and cultured for 30 days before analysis. Gene expression, morphology and wound healing (WH) were analyzed in IPSC-CMs and IPSC-CFs. Results: We observed that IPSC-CFs show higher expression of FLNC than IPSC-CMs (CT CM: 1.04±0.19 vs CT CF: 4.05±0.97, **p<0.01). Besides, FLNC mutated (MT) IPSC-CFs showed lower expression of FLNC when compared to controls (CT), with no changes in expression in IPSC-CMs. Expression of the fibrotic markers POSTN , SMA and THY1 was higher in IPSC-CFs from MT. Immunostaining revealed changes in morphology in IPSC-CFs of MT with accumulation of FLNC in the nucleus. Treatment with TGF-b showed upregulation of CCL2 , SMA , POSTN , COL1A1 and FLNC in MT IPSC-CFs. Wound healing assay revealed no changes in cell migration between CT and MT, although TGF-b treatment increased WH % in MT (MT: 22.5±1.5% vs MT TGF: 59.4±6.5% at 8hs, **p<0.01). Conclusions: Based on our results, we conclude that CFs play an important role on the development of HCM; with higher changes in gene expression, cell migration and morphology than those observed in CMs. Further functional studies will provide evidence of the effects of these changes on the phenotype of the disease.

scholarly journals Thyroid Hormone Status Interferes with Estrogen Target Gene Expression in Breast Cancer Samples in Menopausal Women

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Sandro José Conde ◽  
Renata de Azevedo Melo Luvizotto ◽  
Maria Teresa de Síbio ◽  
Célia Regina Nogueira
Keyword(s):  
Gene Expression ◽  
Thyroid Hormone ◽  
Target Gene ◽  
Progesterone Receptors ◽  
Control Group ◽  
Target Gene Expression ◽  
Menopausal Women ◽  
Hormone Status ◽  
Before And After

We investigated thyroid hormone levels in menopausal BrC patients and verified the action of triiodothyronine on genes regulated by estrogen and by triiodothyronine itself in BrC tissues. We selected 15 postmenopausal BrC patients and a control group of 18 postmenopausal women without BrC. We measured serum TPO-AB, TSH, FT4, and estradiol, before and after surgery, and used immunohistochemistry to examine estrogen and progesterone receptors. BrC primary tissue cultures received the following treatments: ethanol, triiodothyronine, triiodothyronine plus 4-hydroxytamoxifen, 4-hydroxytamoxifen, estrogen, or estrogen plus 4-hydroxytamoxifen. Genes regulated by estrogen (TGFA, TGFB1, and PGR) and by triiodothyronine (TNFRSF9, BMP-6, and THRA) in vitro were evaluated. TSH levels in BrC patients did not differ from those of the control group (1.34 ± 0.60 versus 2.41 ± 1.10 μU/mL), but FT4 levels of BrC patients were statistically higher than controls (1.78 ± 0.20 versus 0.95 ± 0.16 ng/dL). TGFA was upregulated and downregulated after estrogen and triiodothyronine treatment, respectively. Triiodothyronine increased PGR expression; however 4-hydroxytamoxifen did not block triiodothyronine action on PGR expression. 4-Hydroxytamoxifen, alone or associated with triiodothyronine, modulated gene expression of TNFRSF9, BMP-6, and THRA, similar to triiodothyronine treatment. Thus, our work highlights the importance of thyroid hormone status evaluation and its ability to interfere with estrogen target gene expression in BrC samples in menopausal women.

Study the Relationship Between Estrogen Hormone and Calcium in Normal Females Before and After Menopause in Baghdad / Iraq

2018 ◽  
Vol 2 (1) ◽  
Author(s):  
Sameerah Mustafa ◽  
Asal Tawfeeq ◽  
Hadeel Hasan
Keyword(s):  
Oral Contraceptive ◽  
Healthy Controls ◽  
Serum Samples ◽  
Oral Contraceptive Pills ◽  
Contraceptive Pills ◽  
Menopausal Women ◽  
Before And After ◽  
Group A ◽  
Group B ◽  
Age Range

This study involved the collection of (90) samples of women serum which included (30) serum samples collected from women before menopause (reproductive women) in the age range of (22-43) years and were considered as (group A- control). While, (group B) included (30) serum samples collected from women using oral contraceptive pills between the ages of (22-43) years old. Whereas, another (30) serum samples were collected from women after menopause between the ages of (43-54) years and were considered as (group C). All of the collected serum samples were subjected to a number of serological and chemical tests for the measurement of (E2, HDL, LDL and Ca). Then, the obtained data were statistical analyzed and results showed a significant decrease (p˂ 0.05) in (E2 ,Ca and HDL) levels in menopausal women compared to that of the normal healthy controls. While, there were non-significant decrease (p> 0.05) in (E2, Ca and HDL) levels in women taking oral contraceptive when compared to the normal healthy controls. On the other hand, a significant increase (p˂ 0.05) was recorded in LDL level in menopausal women compared to that of the normal healthy controls whereas, no-significant increase (p˃ 0.05) in the LDL level in women taking oral contraceptives when compared to the control women.

scholarly journals Wound Healing Promotion by Hyaluronic Acid: Effect of Molecular Weight on Gene Expression and In Vivo Wound Closure

2021 ◽  
Vol 14 (4) ◽  
pp. 301
Author(s):  
Yayoi Kawano ◽  
Viorica Patrulea ◽  
Emmanuelle Sublet ◽  
Gerrit Borchard ◽  
Takuya Iyoda ◽  
...  
Keyword(s):  
Gene Expression ◽  
Molecular Weight ◽  
Wound Healing ◽  
Hyaluronic Acid ◽  
Healing Process ◽  
Dermal Fibroblasts ◽  
Water Soluble ◽  
Wound Healing Process ◽  
And Migration ◽  
Proliferation And Migration

Hyaluronic acid (HA) has been known to play an important role in wound healing process. However, the effect of molecular weight (MW) of exogenously administered HA on the wound healing process has not been fully understood. In this study, we investigated HA with different MWs on wound healing process using human epidermal keratinocytes and dermal fibroblasts. Cell proliferation and migration ability were assessed by water soluble tetrazolium (WST) assay and wound scratch assay. We examined the effect of HA addition in a full-thickness wound model in mice and the gene expression related to wound healing. Proliferation and migration of HaCaT cells increased with the increase of MW and concentration of HA. Interleukin (IL-1β), IL-8 and vascular endothelial growth factor (VEGF) as well as matrix metalloproteinase (MMP)-9 and MMP-13 were significantly upregulated by high molecular weight (HMW) HA in keratinocytes. Together with VEGF upregulation and the observed promotion of HaCaT migration, HA with the MW of 2290 kDa may hold potential to improve re-epithelialization, a critical obstacle to heal chronic wounds.

scholarly journals Correcting for experiment-specific variability in expression compendia can remove underlying signals

GigaScience ◽  
2020 ◽  
Vol 9 (11) ◽  
Author(s):  
Alexandra J Lee ◽  
YoSon Park ◽  
Georgia Doing ◽  
Deborah A Hogan ◽  
Casey S Greene
Keyword(s):  
Gene Expression ◽  
Large Scale ◽  
Original Signal ◽  
Batch Effects ◽  
Technical Variability ◽  
The Past ◽  
Statistical Correction ◽  
Before And After ◽  
Data Collections ◽  
Biological Patterns

Abstract Motivation In the past two decades, scientists in different laboratories have assayed gene expression from millions of samples. These experiments can be combined into compendia and analyzed collectively to extract novel biological patterns. Technical variability, or "batch effects," may result from combining samples collected and processed at different times and in different settings. Such variability may distort our ability to extract true underlying biological patterns. As more integrative analysis methods arise and data collections get bigger, we must determine how technical variability affects our ability to detect desired patterns when many experiments are combined. Objective We sought to determine the extent to which an underlying signal was masked by technical variability by simulating compendia comprising data aggregated across multiple experiments. Method We developed a generative multi-layer neural network to simulate compendia of gene expression experiments from large-scale microbial and human datasets. We compared simulated compendia before and after introducing varying numbers of sources of undesired variability. Results The signal from a baseline compendium was obscured when the number of added sources of variability was small. Applying statistical correction methods rescued the underlying signal in these cases. However, as the number of sources of variability increased, it became easier to detect the original signal even without correction. In fact, statistical correction reduced our power to detect the underlying signal. Conclusion When combining a modest number of experiments, it is best to correct for experiment-specific noise. However, when many experiments are combined, statistical correction reduces our ability to extract underlying patterns.

scholarly journals T9. EPIGENETIC PROFILING IN SCHIZOPHRENIA DERIVED HUMAN INDUCED PLURIPOTENT STEM CELLS (HIPSCS) AND NEURONS

2020 ◽  
Vol 46 (Supplement_1) ◽  
pp. S234-S234
Author(s):  
Lorna Farrelly ◽  
Shuangping Zhang ◽  
Erin Flaherty ◽  
Aaron Topol ◽  
Nadine Schrode ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mass Spectrometry ◽  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Neural Progenitor Cells ◽  
Gene Expression Pattern ◽  
Early Gene ◽  
Label Free ◽  
Induced Pluripotent

Abstract Background Schizophrenia (SCZ) is a severe psychiatric disorder affecting ~1% of the world’s population. It is largely heritable with genetic risk reflected by a combination of common variants of small effect and highly penetrant rare mutations. Chromatin modifications are known to play critical roles in the mediation of many neurodevelopmental processes, and, when disturbed, may also contribute to the precipitation of psychiatric disorders, such as SCZ. While a handful of candidate-based studies have measured changes in promoter-bound histone modifications, few mechanistic studies have been carried out to explore how these modifications may affect chromatin to precipitate behavioral phenotypes associated with the disease. Methods We applied an unbiased proteomics approach to evaluate the epigenetic landscape of SCZ in human induced pluripotent stem cells (hiPSC), neural progenitor cells (NPCs) and neurons from SCZ patients vs. matched controls. We utilized proteomics-based, label free liquid chromatography mass spectrometry (LC-MS/MS) on purified histones from these cells and confirmed our results by western blotting in postmortem SCZ cortical brain tissues. Furthermore we validated our findings with the application of histone interaction assays and structural and biophysical assessments to identify and confirm novel chromatin ‘readers’. To relate our findings to a SCZ phenotype we used a SCZ rodent model of prepulse inhibition (PPI) to perform pharmacological manipulations and behavioral assessments. Results Using label free mass spectrometry we performed PTM screening of hiPSCs, NPCs and matured neurons derived from SCZ patients and matched controls. We identified, amongst others, altered patterns of hyperacetylation in SCZ neurons. Additionally we identified enhanced binding of particular acetylation ‘reader’ proteins. Pharmacological inhibition of such proteins in an animal model of amphetamine sensitization ameliorated PPI deficits further validating this epigenetic signature in SCZ. Discussion Recent evidence indicates that relevance and patterns of acetylation in epigenetics advances beyond its role in transcription and small molecule inhibitors of these aberrant interactions hold promise as useful therapeutics. This study identifies a role for modulating gene expression changes associated with a SCZ epigenetic signature and warrants further investigation in terms of how this early gene expression pattern perhaps determines susceptibility or severity of the SCZ disease trajectory.

scholarly journals A Prototype Skin Substitute, Made of Recycled Marine Collagen, Improves the Skin Regeneration of Sheep

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1219
Author(s):  
Luca Melotti ◽  
Tiziana Martinello ◽  
Anna Perazzi ◽  
Ilaria Iacopetti ◽  
Cinzia Ferrario ◽  
...  
Keyword(s):  
Gene Expression ◽  
Wound Healing ◽  
Sea Urchin ◽  
Healing Process ◽  
Skin Wound ◽  
Skin Substitute ◽  
Large Animal ◽  
Type I ◽  
Skin Wound Healing ◽  
Skin Adnexa

Skin wound healing is a complex and dynamic process that aims to restore lesioned tissues. Collagen-based skin substitutes are a promising treatment to promote wound healing by mimicking the native skin structure. Recently, collagen from marine organisms has gained interest as a source for producing biomaterials for skin regenerative strategies. This preliminary study aimed to describe the application of a collagen-based skin-like scaffold (CBSS), manufactured with collagen extracted from sea urchin food waste, to treat experimental skin wounds in a large animal. The wound-healing process was assessed over different time points by the means of clinical, histopathological, and molecular analysis. The CBSS treatment improved wound re-epithelialization along with cell proliferation, gene expression of growth factors (VEGF-A), and development of skin adnexa throughout the healing process. Furthermore, it regulated the gene expression of collagen type I and III, thus enhancing the maturation of the granulation tissue into a mature dermis without any signs of scarring as observed in untreated wounds. The observed results (reduced inflammation, better re-epithelialization, proper development of mature dermis and skin adnexa) suggest that sea urchin-derived CBSS is a promising biomaterial for skin wound healing in a “blue biotechnologies” perspective for animals of Veterinary interest.

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