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Pubblicazioni recenti - cardiac fibroblast
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A FOX-Like Mechanism Regulating Lung Fibroblasts - Are We Getting There?
Am J Respir Cell Mol Biol2020 Oct;():. doi: 10.1165/rcmb.2020-0441ED.
Pullamsetti Soni Savai, Günther Andreas,
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Down-regulation of Beclin1 promotes direct cardiac reprogramming.
Sci Transl Med2020 Oct;12(566):. doi: eaay7856.
Wang Li, Ma Hong, Huang Peisen, Xie Yifang, Near David, Wang Haofei, Xu Jun, Yang Yuchen, Xu Yangxi, Garbutt Tiffany, Zhou Yang, Liu Ziqing, Yin Chaoying, Bressan Michael, Taylor Joan M, Liu Jiandong, Qian Li,
Abstract
Direct reprogramming of fibroblasts to alternative cell fates by forced expression of transcription factors offers a platform to explore fundamental molecular events governing cell fate identity. The discovery and study of induced cardiomyocytes (iCMs) not only provides alternative therapeutic strategies for heart disease but also sheds lights on basic biology underlying CM fate determination. The iCM field has primarily focused on early transcriptome and epigenome repatterning, whereas little is known about how reprogramming iCMs remodel, erase, and exit the initial fibroblast lineage to acquire final cell identity. Here, we show that autophagy-related 5 (Atg5)-dependent autophagy, an evolutionarily conserved self-digestion process, was induced and required for iCM reprogramming. Unexpectedly, the autophagic factor Beclin1 (Becn1) was found to suppress iCM induction in an autophagy-independent manner. Depletion of resulted in improved iCM induction from both murine and human fibroblasts. In a mouse genetic model, haploinsufficiency further enhanced reprogramming factor-mediated heart function recovery and scar size reduction after myocardial infarction. Mechanistically, loss of up-regulated and down-regulated Wnt inhibitors, leading to activation of the canonical Wnt/?-catenin signaling pathway. In addition, Becn1 physically interacts with other classical class III phosphatidylinositol 3-kinase (PI3K III) complex components, the knockdown of which phenocopied depletion in cardiac reprogramming. Collectively, our study revealed an inductive role of Atg5-dependent autophagy as well as a previously unrecognized autophagy-independent inhibitory function of Becn1 in iCM reprogramming.
Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.
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68Ga-FAPI-04 Accumulation in Myocardial Infarction in a Patient With Neuroendocrine Carcinoma.
Clin Nucl Med2020 Oct;():. doi: 10.1097/RLU.0000000000003334.
Zhu Wenjia, Guo Fan, Wang Yawen, Ding Haiyan, Huo Li,
Abstract
We present a case of a 64-year-old man with neuroendocrine carcinoma. Incidental findings were demonstrated on Ga-FAPI-04 PET/CT in the inferior wall of left ventricle. A diagnosis of old myocardial infarction was made based on typical electrocardiogram change. Our case suggests that Ga-FAPI PET/CT, as a noninvasive method to reflect fibroblast activation, is potentially feasible for assessment of cardiac remodeling after myocardial infarction in a clinical setting.
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CXADR-like membrane protein protects against heart injury by preventing excessive pyroptosis after myocardial infarction.
J Cell Mol Med2020 Oct;():. doi: 10.1111/jcmm.15955.
Han Xinglong, Zhao Zhen-Ao, Yan Shiping, Lei Wei, Wu Hongchun, Lu Xing-Ai, Chen Yueqiu, Li Jingjing, Wang Yaning, Yu Miao, Wang Yongming, Zheng Yufang, Wang Hongyan, Shen Zhenya, Hu Shijun,
Abstract
Myocardial infarction (MI) results in cardiomyocyte death and ultimately leads to heart failure. Pyroptosis is a type of the inflammatory programmed cell death that has been found in various diseased tissues. However, the role of pyroptosis in MI heart remains unknown. Here, we showed that CXADR-like membrane protein (CLMP) was involved in pyroptosis in the mouse MI heart. Our data showed that CLMP was strongly expressed in fibroblasts of the infarcted mouse hearts. The Clmp mice showed more serious myocardial fibrosis and ventricular dysfunction post-MI than wild-type (Clmp ) mice, indicating a protective effect of the fibroblast-expressed CLMP against MI-induced heart damage. Transcriptome analyses by RNA sequencing indicated that Il-1? mRNA was significantly increased in the MI heart of Clmp mouse, which indicated a more serious inflammatory response. Meanwhile, cleaved caspase-1 and Gasdermin D were significantly increased in the Clmp MI heart, which demonstrated enhanced pyroptosis in the Clmp knockdown heart. Further analysis revealed that the pyroptosis mainly occurred in cardiac fibroblasts (CFs). Compared to wild-type fibroblasts, Clmp CFs showed more serious pyroptosis and inflammatory after LPS plus nigericin treatment. Collectively, our results indicate that CLMP participates in the pyroptotic and inflammatory response of CFs in MI heart. We have provided a novel pyroptotic insight into the ischaemic heart, which might hold substantial potential for the treatment of MI.
© 2020 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.
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Strategies and Challenges to Improve Cellular Programming-Based Approaches for Heart Regeneration Therapy.
Int J Mol Sci2020 Oct;21(20):. doi: E7662.
Jiang Lin, Liang Jialiang, Huang Wei, Wu Zhichao, Paul Christian, Wang Yigang,
Abstract
Limited adult cardiac cell proliferation after cardiovascular disease, such as heart failure, hampers regeneration, resulting in a major loss of cardiomyocytes (CMs) at the site of injury. Recent studies in cellular reprogramming approaches have provided the opportunity to improve upon previous techniques used to regenerate damaged heart. Using these approaches, new CMs can be regenerated from differentiation of iPSCs (similar to embryonic stem cells), the direct reprogramming of fibroblasts [induced cardiomyocytes (iCMs)], or induced cardiac progenitors. Although these CMs have been shown to functionally repair infarcted heart, advancements in technology are still in the early stages of development in research laboratories. In this review, reprogramming-based approaches for generating CMs are briefly introduced and reviewed, and the challenges (including low efficiency, functional maturity, and safety issues) that hinder further translation of these approaches into a clinical setting are discussed. The creative and combined optimal methods to address these challenges are also summarized, with optimism that further investigation into tissue engineering, cardiac development signaling, and epigenetic mechanisms will help to establish methods that improve cell-reprogramming approaches for heart regeneration.
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Endoglin Promotes Myofibroblast Differentiation and Extracellular Matrix Production in Diabetic Nephropathy.
Int J Mol Sci2020 Oct;21(20):. doi: E7713.
Gerrits Tessa, Zandbergen Malu, Wolterbeek Ron, Bruijn Jan A, Baelde Hans J, Scharpfenecker Marion,
Abstract
Diabetic nephropathy (DN) is a complication of diabetes mellitus that can lead to proteinuria and a progressive decline in renal function. Endoglin, a co-receptor of TGF-?, is known primarily for regulating endothelial cell function; however, endoglin is also associated with hepatic, cardiac, and intestinal fibrosis. This study investigates whether endoglin contributes to the development of interstitial fibrosis in DN. Kidney autopsy material from 80 diabetic patients was stained for endoglin and Sirius Red and scored semi-quantitatively. Interstitial endoglin expression was increased in samples with DN and was correlated with Sirius Red staining ( < 0.001). Endoglin expression was also correlated with reduced eGFR ( = 0.001), increased creatinine ( < 0.01), increased systolic blood pressure ( < 0.05), hypertension ( < 0.05), and higher IFTA scores ( < 0.001). Biopsy samples from DN patients were also co-immunostained for endoglin together with CD31, CD68, vimentin, or ?-SMA Endoglin co-localized with both the endothelial marker CD31 and the myofibroblast marker ?-SMA. Finally, we used shRNA to knockdown endoglin expression in a human kidney fibroblast cell line. We found that TGF-?1 stimulation upregulated , , and mRNA and ?-SMA protein, and that these effects were significantly reduced in fibroblasts after endoglin knockdown. Taken together, these data suggest that endoglin plays a role in the pathogenesis of interstitial fibrosis in DN.
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[Research progress on the association between myocardial fibroblast transdifferentiation and diabetic cardiomyopathy].
Zhonghua Xin Xue Guan Bing Za Zhi2020 Oct;48(10):885-889. doi: 10.3760/cma.j.cn112148-20191027-00659.
Gao J M, Hu B, Shen E,
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Unravelling the disease mechanism for TSPYL1 deficiency.
Hum Mol Genet2020 Oct;():. doi: ddaa233.
Buyse Gunnar, Di Michele Michela, Wijgaerts Anouck, Louwette Sophie, Wittevrongel Christine, Thys Chantal, Downes Kate, Ceulemans Berten, Van Esch Hilde, Van Geet Chris, Freson Kathleen,
Abstract
We describe a lethal combined nervous and reproductive systems disease in three affected siblings of a consanguineous family. The phenotype was characterized by visceroautonomic dysfunction (neonatal bradycardia/apnea, feeding problems, hyperactive startle reflex), severe postnatal progressive neurological abnormalities (including abnormal neonatal cry, hypotonia, epilepsy, polyneuropathy, cerebral gray matter atrophy), visual impairment, testicular dysgenesis in males, and sudden death at infant age by brainstem-mediated cardiorespiratory arrest. Whole exome sequencing revealed a novel homozygous frameshift variant p.Val242GlufsTer52 in the TSPY-like 1 gene TSPYL1. The truncated TSPYL1 protein that lacks the nucleosome assembly protein (NAP) domain was retained in the Golgi of fibroblasts from the three patients while control fibroblasts express full length TSPYL1 in the nucleus. Proteomic analysis of nuclear extracts from fibroblasts identified 24 up- and 20 down-regulated proteins in the patients compared to five controls with 'regulation of cell cycle' as the highest scored biological pathway affected. TSPYL1 deficient cells had prolonged S and G2 phases with reduced cellular proliferation rates. Tspyl1 depletion in zebrafish mimicked the patients' phenotype with early lethality, defects in neurogenesis and cardiac dilation. In conclusion, this study reports the third pedigree with recessive TSPYL1 variants, confirming that TSPYL1 deficiency leads to a combined nervous and reproductive systems disease, and provides for the first time insights into the disease mechanism.
© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.
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ANO1 regulates cardiac fibrosis via ATI-mediated MAPK pathway.
Cell Calcium2020 Oct;92():102306. doi: S0143-4160(20)30148-2.
Tian Xiangqin, Sun Changye, Wang Xianwei, Ma Ketao, Chang Yuqiao, Guo Zhikun, Si Junqiang,
Abstract
Cardiac fibrosis is associated with most of heart diseases, but its molecular mechanism remains unclear. Anoctamin-1 (ANO1), a calcium-activated chloride channels (CaCCs) protein, plays a critical role in various pathophysiological processes. In the current study, we identified ANO1 expression in myocardial infarction (MI) model of rat and verified the role of ANO1 in cardiac fibrosis using transcriptomics combined with RNAi assays. we found that ANO1 expression was increased during the first two weeks, and decreased in the third week after MI. Fluorescence double labeling showed that ANO1 was mainly expressed in cardiac fibroblasts (CFs) and displayed an increased expression in CFs with proliferation tendency. The proliferation and secretion of CFs were markedly inhibited by knockdown of ANO1. RNA-Seq showed that most of the downregulation genes were related to the proliferation of CFs and cardiac fibrosis. After ANO1 knockdown, the expressions of angiotensin II type 1 receptor (AT1R) and cell nuclear proliferation antigen were markedly reduced, and the phosphorylation levels of MEK and ERK1/2 was decreased significantly, indicating that ANO1 regulate cardiac fibrosis through ATIR-mediated MAPK signaling pathway. These findings would be useful for the development of therapeutic strategies targeting ANO1 to treat and prevent cardiac fibrosis.
Copyright © 2020 Elsevier Ltd. All rights reserved.
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Extracellular Vesicles of GMSCs Alleviate Aging-Related Cell Senescence.
J Dent Res2020 Oct;():22034520962463. doi: 10.1177/0022034520962463.
Shi H Z, Zeng J C, Shi S H, Giannakopoulos H, Zhang Q Z, Le A D,
Abstract
Healthy aging is a complex biological process with progressive accumulation of senescent cells characterized by stable cell cycle arrest, resulting in impaired homeostasis, regenerative potential, and gradual functional decline in multiple tissues and organs, whereby the aberrant activation of mammalian target of rapamycin (mTOR) signaling networks plays a central role. Herein, we explored the effects of extracellular vesicles (EVs) released by gingiva-derived mesenchymal stem cells (GMSC-EVs) on oxidative stress-induced cellular senescence in human endothelial cells and skin fibroblasts and their antiaging potentials. Our results showed that GMSC-EVs robustly abrogated oxidative stress-induced upregulation in the expression of cellular senescence-related genes, such as ?-galactosidase, p21, p53, and ?H2AX, and mTOR/pS6 signaling pathway, in human umbilical vein endothelial cells (HUVECs) and skin fibroblasts. Meanwhile, GMSC-EVs restored oxidative stress-induced impairment in proliferation and tube formation by HUVECs. Systemic administration of GMSC-EVs attenuated aging-associated elevation in the expression levels of p21, mTOR/pS6, interleukin 6, and tumor necrosis factor ? in skin and heart tissues of aged mice. These findings suggest that GMSC-EVs could be a potential alternative source of cell-free product for attenuation of aging-related skin and vascular dysfunctions due to their potent inhibitory effects on oxidative stress-induced cellular senescence in endothelial cells and skin fibroblasts.
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DPP-4 inhibitor induces FGF21 expression via sirtuin 1 signaling and improves myocardial energy metabolism.
Heart Vessels2020 Oct;():. doi: 10.1007/s00380-020-01711-z.
Furukawa Nozomi, Koitabashi Norimichi, Matsui Hiroki, Sunaga Hiroaki, Umbarawan Yogi, Syamsunarno Mas Rizky A A, Yamaguchi Aiko, Obokata Masaru, Hanaoka Hirofumi, Yokoyama Tomoyuki, Kurabayashi Masahiko,
Abstract
Dipeptidyl peptidase-4 (DPP-4) inhibitors are widely used incretin-based therapy for the treatment of type 2 diabetes. We investigated the cardioprotective effect of a DPP-4 inhibitor, vildagliptin (vilda), on myocardial metabolism and cardiac performance under pressure overload. Mice were treated with either vehicle or vilda, followed by transverse aortic constriction (TAC). After 3 weeks of TAC, cardiac hypertrophy and impairment of systolic function were attenuated in vilda-treated mice. Pressure-volume analysis showed that vilda treatment significantly improved left-ventricular contractile efficiency in TAC heart. Myocardial energy substrate analysis showed that vilda treatment significantly increased glucose uptake as well as fatty acid uptake. Fibroblast growth factor 21 (FGF21), a peptide involved in the regulation of energy metabolism, increased in TAC heart and was further increased by vilda treatment. FGF21 was strongly expressed in cardiac fibroblasts than in cardiomyocytes in mouse heart after TAC with vilda treatment. Vilda treatment markedly induced FGF21 expression in human cardiac fibroblasts through a sirtuin (Sirt) 1-mediated pathway, suggesting that fibroblast-mediated FGF21 expression may regulate energy metabolism and exert vilda-mediated beneficial effects in stressed heart. Vilda induced a metabolic regulator, FGF21 expression in cardiac fibroblasts via Sirt1, and increased contractile efficiency in murine pressure-overloaded heart.
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Effect of Co-culturing Fibroblasts in Human Skeletal Muscle Cell Sheet on Angiogenic Cytokine Balance and Angiogenesis.
Front Bioeng Biotechnol2020 ;8():578140. doi: 10.3389/fbioe.2020.578140.
Thummarati Parichut, Kino-Oka Masahiro,
Abstract
Skeletal muscle comprises a heterogeneous population of myoblasts and fibroblasts. Autologous skeletal muscle myoblasts are transplanted to patients with ischemia to promote cardiac regeneration. In damaged hearts, various cytokines secreted from the skeletal muscle myoblasts promote angiogenesis and consequently the recovery of cardiac functions. However, the effect of skeletal muscle fibroblasts co-cultured with skeletal muscle myoblasts on angiogenic cytokine production and angiogenesis has not been fully understood. To investigate these effects, production of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) was measured using the culture medium of monolayers prepared from various cell densities (mono-culture) and proportions (co-culture) of human skeletal muscle myoblasts (HSMMs) and human skeletal muscle fibroblasts (HSMFs). HSMM and HSMF mono-cultures produced VEGF, whereas HSMF mono-culture produced HGF. The VEGF productivity observed in a monolayer comprising low proportion of HSMFs was two-fold greater than that of HSMM and HSMF mono-cultures. The production of VEGF in HSMMs but not in HSMFs was directly proportional to the cell density. VEGF productivity in non-confluent cells with low cell-to-cell contact was higher than that in confluent cells with high cell-to-cell contact. The dynamic migration of cells in a monolayer was examined to analyze the effect of HSMFs on myoblast-to-myoblast contact. The random and rapid migration of HSMFs affected the directional migration of surrounding HSMMs, which disrupted the myoblast alignment. The effect of heterogeneous populations of skeletal muscle cells on angiogenesis was evaluated using human umbilical vein endothelial cells (HUVECs) incubated with fabricated multilayer HSMM sheets comprising various proportions of HSMFs. Co-culturing HSMFs in HSMM sheet at suitable ratio (30 or 40%) enhances endothelial network formation. These findings indicate the role of HSMFs in maintaining cytokine balance and consequently promoting angiogenesis in the skeletal muscle cell sheets. This approach can be used to improve transplantation efficiency of engineered tissues.
Copyright © 2020 Thummarati and Kino-oka.
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Carcinoid Heart Disease: Pathophysiology, Pathology, Clinical Manifestations, and Management.
Cardiology2020 Oct;():1-9. doi: 10.1159/000507847.
Jin Chengyue, Sharma Ajay Nair, Thevakumar Balasingam, Majid Muhammad, Al Chalaby Shahad, Takahashi Nene, Tanious Ashraf, Arockiam Aro Daniela, Beri Neil, Amsterdam Ezra A,
Abstract
Carcinoid heart disease (CHD) is a rare and potentially lethal manifestation of an advanced carcinoid (neuroendocrine) tumor. The pathophysiology of CHD is related to vasoactive substances secreted by the tumor, of which serotonin is most prominent in the pathophysiology of CHD. Serotonin stimulates fibroblast growth and fibrogenesis, which can lead to cardiac valvular fibrosis. CHD primarily affects right heart valves, causing tricuspid and pulmonic regurgitation and less frequently stenosis of these valves. Left heart valves are usually spared because vasoactive substances such as serotonin are enzymatically inactivated in the lung vasculature. The pathology of CHD is characterized by plaque-like deposition of fibrous tissue on valvular cusps, leaflets, papillary muscles, chordae, and ventricular walls. Symptomatic CHD usually presents between 50 and 70 years of age, initially as dyspnea and fatigue. Echocardiography is the mainstay of imaging and demonstrates thickened right heart valves with limited mobility and regurgitation. Treatment focuses on control of the underlying carcinoid syndrome, targeting subsequent valvular heart disease and managing consequent heart failure. Surgical valve replacement and catheter-directed valve procedures may be effective for selected patients with CHD.
© 2020 S. Karger AG, Basel.
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Polycystin-1, AKT, and CTGF during ischemia/reperfusion.
Biochim Biophys Acta Mol Basis Dis2020 Oct;():165986. doi: S0925-4439(20)30334-3.
Aránguiz P, Romero P, Vásquez F, Flores-Vergara R, Aravena D, Sánchez G, González M, Olmedo I, Pedrozo Z,
Abstract
During ischemia/reperfusion (I/R), cardiomyocytes activate pathways that regulate cell survival and death and release factors that modulate fibroblast-to-myofibroblast differentiation. The mechanisms underlying these effects are not fully understood. Polycystin-1 (PC1) is a mechanosensor crucial for cardiac function. This work aims to assess the role of PC1 in cardiomyocyte survival, its role in profibrotic factor expression in cardiomyocytes, and its paracrine effects on I/R-induced cardiac fibroblast function. In vivo and ex vivo I/R and simulated in vitro I/R (sI/R) were induced in wild-type and PC1-knockout (PC1 KO) mice and PC1-knockdown (siPC1) neonatal rat ventricular myocytes (NRVM), respectively. Neonatal rat cardiac fibroblasts (NRCF) were stimulated with conditioned medium (CM) derived from NRVM or siPC1-NRVM supernatant after reperfusion and fibroblast-to-myofibroblast differentiation evaluated. Infarcts were larger in PC1-KO mice subjected to in vivo and ex vivo I/R, and necrosis rates were higher in siPC1-NRVM than control after sI/R. PC1 activated the pro-survival AKT protein during sI/R and induced PC1-AKT-pathway-dependent CTGF expression. Furthermore, conditioned media from sI/R-NRVM induced PC1-dependent fibroblast-to-myofibroblast differentiation in NRCF. This novel evidence shows that PC1 mitigates cardiac damage during I/R, likely through AKT activation, and regulates CTGF expression in cardiomyocytes via AKT. Moreover, PC1-NRVM regulates fibroblast-to-myofibroblast differentiation during sI/R. PC1, therefore, may emerge as a new key regulator of I/R injury-induced cardiac remodeling.
Copyright © 2020 Elsevier B.V. All rights reserved.
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MMP9 inhibition increases autophagic flux in chronic heart failure.
Am J Physiol Heart Circ Physiol2020 Oct;():. doi: 10.1152/ajpheart.00032.2020.
Nandi Shyam Sundar, Katsurada Kenichi, Sharma Neeru M, Anderson Daniel R, Mahata Sushil K, Patel Kaushik P,
Abstract
An increased matrix metalloprotease9 (MMP9) during post-myocardial infarction (post-MI) exacerbates ischemia-induced chronic heart failure (CHF). Autophagy is cardioprotective during CHF, however, whether increased MMP9 suppressed autophagic activity in CHF is unknown. This study aimed to determine whether the increased MMP9 suppressed autophagic flux, and MMP9 inhibition increased autophagic flux in the heart of rats with Post-MI CHF. Sprague-Dawley rats underwent either Sham or coronary artery ligation, 6-8 weeks prior to being treated with MMP9-inhibitor for 7days, followed by cardiac autophagic flux measurement using lysosomal inhibitor BafilomycinA1. Further, autophagic flux was measured in vitro by treating H9c2 cardiomyocytes with two independent pharmacological MMP9 inhibitors, Salvianolic-Acid-B (SalB) or MMP9-inhibitor-I, and CRISPR/cas9-mediated MMP9 genetic ablation. CHF rats showed cardiac infarct, significantly increased left ventricular end-diastolic pressure (LVEDP), and increased MMP9 activity and fibrosis in the peri-infarct areas of left-ventricular myocardium. The autophagic markers LC3B-II and p62 measurement with lysosomal inhibition showed decreased autophagic flux in the peri-infarct myocardium. Treatment with SalB for 7days in CHF rats decreased MMP9 activity, cardiac fibrosis, and increased autophagic flux in the peri-infarct myocardium. As an in vitro corollary study, measurement of autophagic flux in H9c2 cardiomyocytes and fibroblasts showed that pharmacological inhibition or genetic ablation of MMP9 upregulates autophagic flux. These data are consistent with our observations that MMP9 inhibition upregulates autophagic flux in the heart of rats with CHF. In conclusion, the results in this study suggest that the beneficial outcome of MMP9 inhibition on pathological cardiac remodeling is in part mediated by improved autophagic flux.
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Repurposing mesalazine against cardiac fibrosis in vitro.
Naunyn Schmiedebergs Arch Pharmacol2020 Oct;():. doi: 10.1007/s00210-020-01998-9.
Hoffmann Maximilian, Kant Theresa A, Emig Ramona, Rausch Johanna S E, Newe Manja, Schubert Mario, Künzel Karolina, Winter Luise, Klapproth Erik, Peyronnet Rémi, Ravens Ursula, El-Armouche Ali, Künzel Stephan R,
Abstract
Cardiovascular diseases are exacerbated and driven by cardiac fibrosis. TGF? induces fibroblast activation and differentiation into myofibroblasts that secrete excessive extracellular matrix proteins leading to stiffening of the heart, concomitant cardiac dysfunction, and arrhythmias. However, effective pharmacotherapy for preventing or reversing cardiac fibrosis is presently unavailable. Therefore, drug repurposing could be a cost- and time-saving approach to discover antifibrotic interventions. The aim of this study was to investigate the antifibrotic potential of mesalazine in a cardiac fibroblast stress model. TGF? was used to induce a profibrotic phenotype in a human cardiac fibroblast cell line. After induction, cells were treated with mesalazine or solvent control. Fibroblast proliferation, key fibrosis protein expression, extracellular collagen deposition, and mechanical properties were subsequently determined. In response to TGF? treatment, fibroblasts underwent a profound phenoconversion towards myofibroblasts, determined by the expression of fibrillary ?SMA. Mesalazine reduced differentiation nearly by half and diminished fibroblast proliferation by a third. Additionally, TGF? led to increased cell stiffness and adhesion, which were reversed by mesalazine treatment. Collagen 1 expression and deposition-key drivers of fibrosis-were significantly increased upon TGF? stimulation and reduced to control levels by mesalazine. SMAD2/3 and ERK1/2 phosphorylation, along with reduced nuclear NF?B translocation, were identified as potential modes of action. The current study provides experimental pre-clinical evidence for antifibrotic effects of mesalazine in an in vitro model of cardiac fibrosis. Furthermore, it sheds light on possible mechanisms of action and suggests further investigation in experimental and clinical settings.
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Angiogenesis After Acute Myocardial Infarction.
Cardiovasc Res2020 Oct;():. doi: cvaa287.
Wu Xuekun, Reboll Marc R, Korf-Klingebiel Mortimer, Wollert Kai C,
Abstract
Acute myocardial infarction (MI) inflicts massive injury to the coronary microcirculation leading to vascular disintegration and capillary rarefication in the infarct region. Tissue repair after MI involves a robust angiogenic response that commences in the infarct border zone and extends into the necrotic infarct core. Technological advances in several areas have provided novel mechanistic understanding of postinfarction angiogenesis and how it may be targeted to improve heart function after MI. Cell lineage tracing studies indicate that new capillary structures arise by sprouting angiogenesis from preexisting endothelial cells (ECs) in the infarct border zone with no meaningful contribution from non-endothelial cell sources. Single cell RNA sequencing (scRNAseq) shows that ECs in infarcted hearts may be grouped into clusters with distinct gene expression signatures, likely reflecting functionally distinct cell populations. EC-specific multicolor lineage tracing reveals that EC subsets clonally expand after MI. Expanding EC clones may arise from tissue-resident ECs with stem cell characteristics that have been identified in multiple organs including the heart. Tissue repair after MI involves interactions among multiple cell types which occur, to a large extent, through secreted proteins and their cognate receptors. While we are only beginning to understand the full complexity of this intercellular communication, macrophage and fibroblast populations have emerged as major drivers of the angiogenic response after MI. Animal data support the view that the endogenous angiogenic response after MI can be boosted to reduce scarring and adverse left ventricular remodeling. The improved mechanistic understanding of infarct angiogenesis therefore creates multiple therapeutic opportunities. During preclinical development, all proangiogenic strategies should be tested in animal models that replicate both cardiovascular risk factor(s) and the pharmacotherapy typically prescribed to patients with acute MI. Considering that the majority of patients nowadays do well after MI, clinical translation will require careful selection of patients in need of proangiogenic therapies.
Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2020. For permissions please email: journals.permissions@oup.com.
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Unique patterns of cardiogenic and fibrotic gene expression in rat cardiac fibroblasts.
Vet World2020 Aug;13(8):1697-1708. doi: 10.14202/vetworld.2020.1697-1708.
Tachampa Kittipong, Wongtawan Tuempong,
Abstract
Background and Aim:
Cardiac fibroblasts are important for both normal and pathological states of the heart, but the knowledge in cell physiology and genomics is still poorly understood. The aims of the present study were; first, to investigate the expression of cardiac and fibrotic genes in rat cardiac fibroblasts compared to cardiomyocytes and other fibroblasts (skin and muscle fibroblasts), second, to examine the in vitro effect of serum concentration on fibroblast gene expression. The findings can potentially be applied in ischemia/reperfusion models.
Materials and Methods:
Rat cardiac fibroblasts were collected and cultured in different conditions, and their gene expression (21 cardiogenic genes and 16 fibrotic genes) was compared with cardiomyocytes and other fibroblasts using comparative quantitative polymerase chain reaction. We also mimicked myocardial ischemia/reperfusion by depleting and then adding a serum into the culture in conventional culture (10% serum).
Results:
Cardiac fibroblasts expressed most of the cardiogenic genes, but their expression levels were significantly lower than in cardiomyocytes, while almost all fibrotic genes in the cardiac fibroblasts were significantly more highly expressed than in cardiomyocytes, except matrix metallopeptidase 9 (Mmp9) which also had greater expression in other fibroblasts. After mimicking cardiac ischemia and reperfusion in vitro by starving and then adding a serum into the cardiac fibroblast culture, the results revealed that Mmp9 expression was significantly increased (>30 times) after increasing but not reducing the serum in the culture. The expression of most cardiogenic and fibrotic genes in cardiac fibroblasts tended to decrease after increasing the serum in the culture. These changes were specific to cardiac fibroblasts but no other fibroblasts.
Conclusion:
Cardiac fibroblasts have a distinct pattern of gene expression from other fibroblasts and cardiomyocytes. They are also sensitive to high serum concentration but not affected by serum depletion, suggesting that the process of developing cardiac fibrosis might be stimulated by reperfusion or overcirculation rather than ischemia. The cell starvation followed the adding of serum may serve as a useful model to study cardiac fibrosis cause by the change of blood flow.
Copyright: © Tachampa and Wongtawan.
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An3D diabetic human skin model from diabetic primary cells.
Biomed Mater2020 Oct;():. doi: 10.1088/1748-605X/abc1b1.
Yilmaz Ozdogan Candan, Kenar Halime, Davun Kivanc Emre, Yucel Deniz, Doger Emek, Alagoz Sahin,
Abstract
Diabetes mellitus, a complex metabolic disorder, leads to many health complications like kidney failure, diabetic heart disease, stroke, and foot ulcers. Treatment approaches of diabetes and identification of the mechanisms underlying diabetic complications of the skin have gained importance due to continued rapid increase in the diabetes incidence. A thick and pre-vascularized in vitro 3D type 2 diabetic human skin model (DHSM) was developed in this study. The methacrylated gelatin (GelMA) hydrogel was produced by photocrosslinking and its pore size (54.85±8.58 ?m), compressive modulus (4.53±0.67 kPa) and swelling ratio (17.5±2.2 %) were found to be suitable for skin tissue engineering. 8% GelMA hydrogel effectively supported the viability, spreading and proliferation of human dermal fibroblasts. By isolating dermal fibroblasts, HUVECs and keratinocytes from type 2 diabetic patients, an in vitro 3D type 2 diabetic human skin model, 12 mm in width and 1.86 mm thick, was constructed. The skin model consisted of a continuous basal epidermal layer and a dermal layer with blood capillary-like structures, ideal for evaluating the effects of anti-diabetic drugs and wound healing materials and factors. The functionality of the DHSM was showed by applying a therapeutic hydrogel into its central wound; especially fibroblast migration to the wound site was evident in 9 days. We have demonstrated that DHSM is a biologically relevant model with sensitivity and predictability in evaluating the diabetic wound healing potential of a therapeutic material.
© 2020 IOP Publishing Ltd.
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Germline and Mosaic Variants in PRKACA and PRKACB Cause a Multiple Congenital Malformation Syndrome.
Am J Hum Genet2020 Oct;():. doi: S0002-9297(20)30325-6.
Palencia-Campos Adrian, Aoto Phillip C, Machal Erik M F, Rivera-Barahona Ana, Soto-Bielicka Patricia, Bertinetti Daniela, Baker Blaine, Vu Lily, Piceci-Sparascio Francesca, Torrente Isabella, Boudin Eveline, Peeters Silke, Van Hul Wim, Huber Celine, Bonneau Dominique, Hildebrand Michael S, Coleman Matthew, Bahlo Melanie, Bennett Mark F, Schneider Amy L, Scheffer Ingrid E, Kibćk Maria, Kristiansen Britta S, Issa Mahmoud Y, Mehrez Mennat I, Ismail Samira, Tenorio Jair, Li Gaoyang, Skĺlhegg Bjřrn Steen, Otaify Ghada A, Temtamy Samia, Aglan Mona, Jřnch Aia E, De Luca Alessandro, Mortier Geert, Cormier-Daire Valérie, Ziegler Alban, Wallis Mathew, Lapunzina Pablo, Herberg Friedrich W, Taylor Susan S, Ruiz-Perez Victor L,
Abstract
PRKACA and PRKACB code for two catalytic subunits (C? and C?) of cAMP-dependent protein kinase (PKA), a pleiotropic holoenzyme that regulates numerous fundamental biological processes such as metabolism, development, memory, and immune response. We report seven unrelated individuals presenting with a multiple congenital malformation syndrome in whom we identified heterozygous germline or mosaic missense variants in PRKACA or PRKACB. Three affected individuals were found with the same PRKACA variant, and the other four had different PRKACB mutations. In most cases, the mutations arose de novo, and two individuals had offspring with the same condition. Nearly all affected individuals and their affected offspring shared an atrioventricular septal defect or a common atrium along with postaxial polydactyly. Additional features included skeletal abnormalities and ectodermal defects of variable severity in five individuals, cognitive deficit in two individuals, and various unusual tumors in one individual. We investigated the structural and functional consequences of the variants identified in PRKACA and PRKACB through the use of several computational and experimental approaches, and we found that they lead to PKA holoenzymes which are more sensitive to activation by cAMP than are the wild-type proteins. Furthermore, expression of PRKACA or PRKACB variants detected in the affected individuals inhibited hedgehog signaling in NIH 3T3 fibroblasts, thereby providing an underlying mechanism for the developmental defects observed in these cases. Our findings highlight the importance of both C? and C? subunits of PKA during human development.
Copyright © 2020 American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.
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