AIMS:With the extensive application of immunotherapy in treating cancer, the immunotherapy-related cardiovascular toxicity (ITR-CVT) has gotten a rapid recognition due to its high mortality. Previously, we have found that potential cancer immunotherapies based on promoting iNKT cell activation exacerbate ISO-induced cardiac injury, but the underlying mechanism is unknown. The current study is to determine which specific cell type/s and the corresponding molecular pathways are responsible for such a cardiotoxicity. MATERIALS AND METHODS:Transcriptome sequencing and bioinformatic analysis were performed on heart tissues from an enhanced cardiac injury model following iNKT cell activation via α-Galactosylceramide (αGC). The role of IFN-γ-STAT1 signaling was validated using IFN-γ antibody blocking and JAK-STAT1 chemical inhibition. The experiments of Macrophage isolation and depletion were conducted to assess cell-specific contributions. In vitro co-culture experiments with αGC-primed macrophages and fibroblasts were conducted under STAT1 inhibition or silencing. Tumor-bearing mice were also examined. KEY FINDINGS:Transcriptome analysis identified IFN-γ-STAT1 signaling as central to the enhanced cardiac injury, blocking IFN-γ or inhibiting STAT1 could attenuate the injury. Macrophages were identified as the main source of IFN-γ-STAT1 activation, and their depletion significantly reversed cardiac injury exacerbation. In vitro, STAT1 inhibition or silencing reduced fibroblast activation induced by αGC-primed macrophages. In tumor-bearing mice, αGC also further exacerbated cardiac injury. SIGNIFICANCE:These findings revealed that the activation of STAT1 in cardiac macrophages via IFNγ critically contributes to cardiotoxicity induced by iNKT-immunotherapy, which provides a potential method to manage ITR-CVT in patients.

With an in-depth understanding of cardiac cell differentiation, cell therapy derived from stem cells has shown promising therapeutic effects in the treatment of myocardial infarction (MI). Although many types of cardiac or noncardiac cells have been found to play protective roles in MI, the specific role of endocardial cells (ECCs) in MI has not been reported. The current study was designed to determine whether human embryonic stem cell (hESC)-derived endocardial cells (hESC-ECCs) could be protective against MI. We first developed a cell delivery system by constructing a photosensitive interpenetrating network hydrogel consisting of gelatin methacryloyl (GelMA) and silk fibroin methacryloyl (SilMA). The sorted hESC-ECCs were loaded into the delivery system and then injected into the pericardium cavity of the MI rats. These results show that the cell delivery system has good biocompatibility. Moreover, the delivered endocardial cells improved cardiac function and delayed capillary atrophy after MI. Further mechanistic analysis revealed that hESC-ECCs protect the mitochondria of cardiomyocytes from damage under oxidative stress and potentially promote the angiogenesis of cardiac endothelial cells. Our results demonstrated that hESC-ECCs have the potential to serve as a cell therapy strategy for MI treatment by maintaining cardiomyocyte survival and facilitating angiogenesis.

It has recently become more recognized that renal diseases in adults can originate from adverse intrauterine (maternal) environmental exposures. Previously, we found that prenatal lipopolysaccharide (LPS) exposure can result in chronic renal inflammation, which leads to renal damage in older offspring rats. To test whether prenatal inflammatory exposure predisposes offspring to renal damage, a mouse model of oral adenine consumption-induced chronic kidney disease (CKD) was applied to offspring from prenatal LPS-treated mothers (offspring-pLPS) and age-matched control offspring of prenatal saline-treated mothers (offspring-pSaline). We found that offspring-pLPS mice presented with more severe renal collagen deposition and renal dysfunction after 4 weeks of adenine consumption than sex- and treatment-matched offspring-pSaline controls. To illustrate the underlying molecular mechanism, we subjected offspring-pLPS and offspring-pSaline kidneys to genome-wide transcriptomic analysis. Bioinformatic analysis of the sequencing data, together with further experimental confirmation, revealed a strong activation of the PERK-eIF2α-ATF4-mediated unfolded protein response (UPR) in offspring-pLPS kidneys, which likely contributed to the CKD predisposition seen in offspring-pLPS mice. More importantly, the specific eIF2α-ATF4 signaling inhibitor ISIRB was able to prevent adenine-induced CKD in the offspring-pLPS mice. Our findings suggest that the eIF2α-ATF4-mediated UPR, but not PERK, is likely the major disease-causing pathway in prenatal inflammatory exposure-induced CKD predisposition. Our study also suggests that targeting this signaling pathway is a potentially promising approach for CKD treatment.

Altered cardiac innate immunity is highly associated with the progression of cardiac disease states and heart failure. S100A8/A9 is an important component of damage-associated molecular patterns (DAMPs) that is critically involved in the pathogenesis of heart failure, thus considered a promising target for pharmacological intervention. In the current study, initially, we validated the role of S100A8/A9 in contributing to cardiac injury and heart failure via the overactivation of the β-adrenergic pathway and tested the potential use of paquinimod as a pharmacological intervention of S100A8/A9 activation in preventing cardiac dysfunction, collagen deposition, inflammation, and immune cell infiltration in β-adrenergic overactivation–mediated heart failure. This finding was further confirmed by the cardiomyocyte-specific silencing of S100A9 via the use of the adeno-associated virus (AAV) 9-mediated short hairpin RNA (shRNA) gene silencing system. Most importantly, in the assessment of the underlying cellular mechanism by which activated S100A8/A9 cause aggravated progression of cardiac fibrosis and heart failure, we discovered that the activated S100A8/A9 can promote fibroblast-macrophage interaction, independent of inflammation, which is likely a key mechanism leading to the enhanced collagen production. Our results revealed that targeting S100A9 provides dual beneficial effects, which is not only a strategy to counteract cardiac inflammation but also preclude cardiac fibroblast-macrophage interactions. The findings of this study also indicate that targeting S100A9 could be a promising strategy for addressing cardiac fibrosis, potentially leading to future drug development.

HomeCirculationVol. 145, No. 13Activation of iNKT Cells at the Maternal–Fetal Interface Predisposes Offspring to Cardiac Injury Free AccessLetterPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toFree AccessLetterPDF/EPUBActivation of iNKT Cells at the Maternal–Fetal Interface Predisposes Offspring to Cardiac Injury Dayan Cao, MD, PhD, Ying Liu, PhD, Xin Chen, PhD, Jie Liu, MS, Jie Liu, MS, Wenjing Lai, MS, Shuhui Li, MD, PhD, Wenjia Wang, MS, Wenjun Zhang, MD, Deyong Xiao, PhD, Kai Yang, PhD, Baiyan Li, MD, PhD, Zhongjun Zhou, PhD, Chen-Leng Cai, PhD, Xiaohui Li, MD, PhD and Weinian Shou, PhD Dayan CaoDayan Cao https://orcid.org/0000-0001-7648-3668 Institute of Materia Medica and Center of Translational Medicine (D.C., J.L., J.L., X.C., W.W., X.L.), Army Medical University, Chongqing, China. Search for more papers by this author , Ying LiuYing Liu https://orcid.org/0000-0001-9447-030X Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis (Y.L., W.Z., D.X., K.Y., B.L., C.C., W.S.). Search for more papers by this author , Xin ChenXin Chen Institute of Materia Medica and Center of Translational Medicine (D.C., J.L., J.L., X.C., W.W., X.L.), Army Medical University, Chongqing, China. Search for more papers by this author , Jie LiuJie Liu https://orcid.org/0000-0002-8353-5582 Institute of Materia Medica and Center of Translational Medicine (D.C., J.L., J.L., X.C., W.W., X.L.), Army Medical University, Chongqing, China. Search for more papers by this author , Jie LiuJie Liu https://orcid.org/0000-0002-8353-5582 Institute of Materia Medica and Center of Translational Medicine (D.C., J.L., J.L., X.C., W.W., X.L.), Army Medical University, Chongqing, China. Search for more papers by this author , Wenjing LaiWenjing Lai Department of Pharmacy, Xinqiao Hospital (W.L.), Army Medical University, Chongqing, China. Search for more papers by this author , Shuhui LiShuhui Li Department of Clinical Biochemistry, College of Pharmacy (S.L.), Army Medical University, Chongqing, China. Search for more papers by this author , Wenjia WangWenjia Wang Institute of Materia Medica and Center of Translational Medicine (D.C., J.L., J.L., X.C., W.W., X.L.), Army Medical University, Chongqing, China. Search for more papers by this author , Wenjun ZhangWenjun Zhang https://orcid.org/0000-0001-9801-0634 Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis (Y.L., W.Z., D.X., K.Y., B.L., C.C., W.S.). Search for more papers by this author , Deyong XiaoDeyong Xiao https://orcid.org/0000-0002-8353-5582 Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis (Y.L., W.Z., D.X., K.Y., B.L., C.C., W.S.). Search for more papers by this author , Kai YangKai Yang Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis (Y.L., W.Z., D.X., K.Y., B.L., C.C., W.S.). Search for more papers by this author , Baiyan LiBaiyan Li Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis (Y.L., W.Z., D.X., K.Y., B.L., C.C., W.S.). Search for more papers by this author , Zhongjun ZhouZhongjun Zhou School of Biomedical Sciences, Faculty of Medicine, The University of Hong Kong, China (Z.Z.). Search for more papers by this author , Chen-Leng CaiChen-Leng Cai https://orcid.org/0000-0001-6900-4737 Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis (Y.L., W.Z., D.X., K.Y., B.L., C.C., W.S.). Search for more papers by this author , Xiaohui LiXiaohui Li Xiaohui Li, MD, PhD, Institute of Materia Medica and Center of Translational Medicine, College of Pharmacy, Army Medical University, No 30 Gaotanyan Road, Shapingba District, Chongqing, 400038, China. Email E-mail Address: [email protected] https://orcid.org/0000-0001-9653-9227 Institute of Materia Medica and Center of Translational Medicine (D.C., J.L., J.L., X.C., W.W., X.L.), Army Medical University, Chongqing, China. Search for more papers by this author and Weinian ShouWeinian Shou Correspondence to: Weinian Shou, PhD, Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, 1044 West Walnut Street, Indianapolis, IN 46202; Email E-mail Address: [email protected] https://orcid.org/0000-0001-5538-9605 Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis (Y.L., W.Z., D.X., K.Y., B.L., C.C., W.S.). Search for more papers by this author Originally published28 Mar 2022https://doi.org/10.1161/CIRCULATIONAHA.121.054239Circulation. 2022;145:1032–1035The fetal origin of adult disease (FOAD) paradigm is closely linked to the onset of cardiovascular disorders. Epidemiologic studies have suggested that intrauterine inflammatory environments are likely associated with chronic cardiovascular and metabolic diseases.1 We previously demonstrated that administration of a low dose of lipopolysaccharide (LPS) to pregnant mice did not grossly alter pregnancy or overall fetal development but could trigger cardiovascular events in 8-month-old offspring (offspring–prenatal LPS [pLPS]), thus establishing a unique FOAD experimental model (Figure [A]).2Download figureDownload PowerPointFigure. Activation of invariant natural killer T cells at the maternal–fetal interface predisposes offspring to cardiac injury. A, Schematic diagram of the experimental design of the prenatal low-dose lipopolysaccharide (LPS) exposure (Sigma; 75 μg/kg body weight) at 11.5 dpc and the administration of isoproterenol (ISO; Sigma; 5 mg/kg body weight) in 4-week-old offspring. B, Fluorescence-activated cell sorting analysis of the proportions of major immune cell types in offspring–prenatal lipopolysaccharide (pLPS) hearts and age-matched offspring–prenatal saline (pSaline) hearts, including T cells (CD45+CD19–CD3+), B cells (CD45+CD19+CD3–), dendritic cells (DCs; CD45+CD3–CD19–Ly6G–CD11b+F4/80–CD11c+MHCII+), and granulocytes (CD45+CD3–CD19–Ly6G+CD11b+). C, Fluorescence-activated cell sorting analysis of macrophages and monocyte-derived macrophages in offspring-pLPS hearts (4 weeks old) and age-matched offspring-pSaline hearts. Gating strategy is as indicated. D, Mammalian phenotype ontology analysis using the R package GSVA (gene set variation analysis). The data are presented as a heatmap and the original RNA sequencing data were deposited to the National Center for Biotechnology Information Sequence Read Archive (https://dataview.ncbi.nlm.nih.gov/object/PRJNA742378). E, Immunohistologic analyses of isoproterenol-mediated heart injury. The light brown (white arrows) or dark brown (red arrows) signals are positive staining signals. F, Mammalian phenotype ontology enrichment analysis presented as a heatmap reveals hyperactivation of inflammatory pathways in isoproterenol-treated offspring-pLPS hearts. The original RNA sequencing data were deposited to the Sequence Read Archive (https://dataview.ncbi.nlm.nih.gov/object/PRJNA742378). G, Fluorescence-activated cell sorting analysis of invariant natural killer T (iNKT) cells in the placenta from 48 to 96 hours after prenatal LPS exposure. H, Fluorescence-activated cell sorting analyses of iNKT activation in the uterus, decidua, and placenta at 72 hours after prenatal LPS exposure. The spleen serves as the positive control. I, A schematic diagram of the experimental design using CD1d-deficient female mice to assess whether maternal iNKT cell activation is essential for the enhanced cardiac injury in prenatally inflammatory stimulated offspring. J, Immunohistologic analysis of heart injury. Maternal iNKT cell deficiency abolishes the hypersensitivity to cardiac injury phenotype in offspring hearts. K, The top-ranked molecular networks identified by ingenuity pathway analysis in fetal liver-pLPS versus fetal liver-pSaline at 14.5 dpc. The original RNA sequencing data were deposited to the Sequence Read Archive (https://dataview.ncbi.nlm.nih.gov/object/PRJNA742673). L, Western blot analysis confirmed the downregulation of GFI1 (growth factor independent 1 transcriptional repressor) protein expression in fetal liver-pLPS, further indicating potentially altered myelopoiesis. M, A schematic diagram summarizing the potential pathogenetic pathway by which maternal inflammatory exposure predisposes the offspring hearts to injury. All quantification data are expressed as mean±SD; statistical analysis was performed with the Student independent t test or 1-way analysis of variance with the Tukey test. P<0.05 is considered significant.To explore the underlying mechanism, we quantified major immune cell types in 4-week-old offspring-pLPS hearts by fluorescence-activated cell sorting analysis (Figure [B and C]) and did not find significant alterations in these cell types except CCR2+ (C-C chemokine receptor type 2) macrophages. The proportional and normalized cell numbers of CCR2+ macrophages were significantly increased in offspring-pLPS compared with offspring–prenatal saline (offspring-pSaline; Figure [C]). CCR2+ macrophages are associated with the cardiac hyperinflammatory response. Genome-wide transcriptomic analysis demonstrated 105 differentially expressed genes in offspring-pLPS hearts, and mammalian phenotype ontology term analysis revealed that these altered genes are associated with abnormal cardiac immunity (Figure [D]), supporting the finding of increased levels of infiltrated macrophages in offspring-pLPS hearts (Figure [C]). We further tested whether the altered cardiac immunity predisposes offspring-pLPS hearts to injury (Figure [A]). As shown in Figure (E), although the levels of cardiac fibrosis remained low in both 4-week-old offspring-pLPS and offspring-pSaline hearts before isoproterenol treatment, the infusion of isoproterenol significantly induced more severe degrees of cardiac fibrosis and dramatically increased levels of infiltrated immune cells and macrophages in isoproterenol-treated offspring-pLPS hearts compared with isoproterenol-treated offspring-pSaline hearts. Isoproterenol treatment led to 440 upregulated genes and 62 downregulated genes in offspring-pLPS hearts. These upregulated genes were highly enriched in the enhanced level of inflammatory response and cell death pathways in offspring-pLPS/isoproterenol hearts compared with offspring-pSaline/isoproterenol hearts (Figure [F]). These data collectively demonstrate that prenatal inflammatory exposure can lead to altered cardiac innate immunity and predispose the offspring heart to injury.Placental tissues are rich in immune cells.3 Given that LPS treatment can lead to placental injury,4 we reasoned that the immune response at the maternal–fetal interface is potentially the main driving factor of this effect. To determine the immune response in the placenta, we analyzed the relative proportional changes in immune cell types, including T cells, B cells, macrophages, natural killer (NK) cells, and invariant natural killer T (iNKT) cells, in the placenta. Under our low-dose LPS conditions, we found that NK cells (data not shown) and iNKT cells were significantly increased (Figure [G]). NK cells are the major immune cells that play critical roles in mediating placental immune tolerance and immunomodulation. The increase in NK cells was expected. However, iNKT cells, a minor immune cell population in the placenta, were also significantly upregulated at 72 hours after LPS treatment. Given that iNKT cells have a long-lasting effect of preserving inflammatory activity and are activated by LPS, we thus performed a more detailed analysis on relative iNKT and activated iNKT cell numbers at the maternal–fetal interface in response to prenatal LPS treatment. Significant iNKT accumulation was first noted in the uterus at 48 hours after prenatal LPS exposure (data not shown) followed by a dramatic increase in the decidua and placenta at 72 hours (Figure [H], left panel). Consistent with this finding, activated iNKT cells were enriched in the decidua and placenta (Figure [H], right panel). To determine whether maternal iNKT cell activation is essential to the hypersensitivity to injury phenotype in offspring hearts, we used iNKT cell-deficient CD1d-knockout pregnant mice administered LPS at 11.5 dpc followed by isoproterenol treatment of 4-week-old offspring (Figure [I]). We found that in CD1d-knockout mice, the hypersensitivity to injury phenotype was abolished in offspring hearts, and comparable levels of cardiac fibrosis and infiltration of immune and CCR2+ macrophages were noted between isoproterenol-treated offspring-pLPS and offspring-pSaline (Figure [J]).Hematopoiesis in the fetal liver serves as a key sensor of maternal infection in modulating the development of the fetal immune system.5 We performed transcriptomic analysis of fetal livers and found a total of 101 differentially expressed genes in fetal liver-pLPS compared with fetal liver-pSaline. These genes are associated with networks in the development of the hematopoietic system, especially myeloid cells (Figure [K]). GFI1 (growth factor independent 1 transcriptional repressor) is a critical transcriptional repressor that regulates myeloid lineage differentiation and is downregulated in fetal liver-pLPS (Figure [L]). Anomalous myelopoiesis in fetal liver-pLPS is likely a key contributor to exacerbating monocyte-derived infiltrated macrophages in offspring-pLPS hearts. Our data collectively document a critical role of maternal iNKT cells in mediating prenatal inflammatory exposure and promoting the hypersensitivity to injury phenotype in offspring hearts (Figure [M]).All animal studies conformed to the Public Health Service Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committee of Indiana University and Army Medical University. The original data and research materials are available on request. The original RNA sequencing data were deposited in the National Center for Biotechnology Information Sequence Read Archive (URL: https://www.ncbi.nlm.nih.gov/sra).Article InformationAcknowledgmentsThe authors thank the members of Dr Shou’s and Dr X. Li’s laboratories for reading the manuscript and providing comments.Sources of FundingThis work is supported in part by the Riley Children Foundation (to Dr Shou), National Natural Science Foundation of China (numbers 81520108029, 81773742, and 81473210 to Dr X. Li), and [email protected] Program (to Dr Zhou).Nonstandard Abbreviations and AcronymsCCR2C-C chemokine receptor type 2FOADfetal origin of adult diseaseGFI1growth factor independent 1 transcriptional repressoriNKTinvariant natural killer TLPSlipopolysaccharideNKnatural killerpLPSprenatal lipopolysaccharide treatedpSalineprenatal saline treatedDisclosures None.FootnotesFor Sources of Funding and Disclosures, see page 1034.https://www.ahajournals.org/journal/circCorrespondence to: Weinian Shou, PhD, Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, 1044 West Walnut Street, Indianapolis, IN 46202; Email [email protected]eduXiaohui Li, MD, PhD, Institute of Materia Medica and Center of Translational Medicine, College of Pharmacy, Army Medical University, No 30 Gaotanyan Road, Shapingba District, Chongqing, 400038, China. Email [email protected]comReferences1. Mazumder B, Almond D, Park K, Crimmins EM, Finch CE. Lingering prenatal effects of the 1918 influenza pandemic on cardiovascular disease.J Dev Orig Health Dis. 2010; 1:26–34. doi: 10.1017/S2040174409990031CrossrefMedlineGoogle Scholar2. Deng Y, Song L, Nie X, Shou W, Li X. Prenatal inflammation exposure-programmed cardiovascular diseases and potential prevention.Pharmacol Ther. 2018; 190:159–172. doi: 10.1016/j.pharmthera.2018.05.009CrossrefMedlineGoogle Scholar3. Erlebacher A. Immunology of the maternal-fetal interface.Annu Rev Immunol. 2013; 31:387–411. doi: 10.1146/annurev-immunol-032712-100003CrossrefMedlineGoogle Scholar4. Fricke EM, Elgin TG, Gong H, Reese J, Gibson-Corley KN, Weiss RM, Zimmerman K, Bowdler NC, Kalantera KM, Mills DA, et al.. Lipopolysaccharide-induced maternal inflammation induces direct placental injury without alteration in placental blood flow and induces a secondary fetal intestinal injury that persists into adulthood.Am J Reprod Immunol. 2018; 79:e12816. doi: 10.1111/aji.12816CrossrefMedlineGoogle Scholar5. Apostol AC, Jensen KDC, Beaudin AE. Training the fetal immune system through maternal inflammation-a layered hygiene hypothesis.Front Immunol. 2020; 11:123. doi: 10.3389/fimmu.2020.00123CrossrefMedlineGoogle Scholar Previous Back to top Next FiguresReferencesRelatedDetails March 29, 2022Vol 145, Issue 13 Article InformationMetrics © 2022 American Heart Association, Inc.https://doi.org/10.1161/CIRCULATIONAHA.121.054239PMID: 35344410 Originally publishedMarch 28, 2022 Keywordsinflammationheart failureimmunityPDF download Advertisement SubjectsBasic Science ResearchInflammationMyocardial Biology

Phillyrin, a well-known natural compound from the dried fruits of Forsythia suspensa (Thunb.) Vahl., has shown anti-inflammatory, antioxidant and anti-virus activities as well as renal protective effects on diabetic nephropathy. In this study, we investigated whether phillyrin attenuated cardiac hypertrophy induced by catecholamine in vivo and in vitro, and explored the underlying mechanisms. Cardiac hypertrophy was induced in C57BL/6 mice by subcutaneous injection of norepinephrine (NE, a key catecholamine), and in rat cardiomyoblasts (H9c2) by stimulation with NE in vitro. Our results showed that administration of phillyrin (100 mg/kg, i.p. for 15 days) significantly improved cardiac function, histopathological changes, cardiac hypertrophy and decreased the upregulated hypertrophic markers (ANP, BNP, and 13-MHC). Moreover, treatment with phillyrin obviously reduced the infiltration of the CD68 positive macrophages and the mRNA expression of proinflammatory genes (IL-113, IL-6, and TNF-alpha) in left ventricular tissue. In addition, treatment with phillyrin markedly inhibited the phosphorylation of p38 MAPK, ERK1/2, AKT, and NF-xB p65 in heart tissues. Furthermore, in NE-treated H9c2 cells, pretreatment with phillyrin clearly attenuated cardiomyocyte hypertrophy, reduced ROS production and inhibited the phosphorylation of p38 MAPK, ERK1/2, AKT, and NF-xB p65 in vitro. Collectively, our results demonstrate that phillyrin effectively alleviates NE-induced cardiac hypertrophy and inflammatory response by suppressing p38 MAPK/ERK1/2 and AKT/NF-xB signaling pathways.

The RNA-binding protein QKI belongs to the hnRNP K-homology domain protein family, a well-known regulator of pre-mRNA alternative splicing and is associated with several neurodevelopmental disorders. Qki is found highly expressed in developing and adult hearts. By employing the human embryonic stem cell (hESC) to cardiomyocyte differentiation system and generating QKI-deficient hESCs (hESCs-QKIdel) using CRISPR/Cas9 gene editing technology, we analyze the physiological role of QKI in cardiomyocyte differentiation, maturation, and contractile function. hESCs-QKIdel largely maintain normal pluripotency and normal differentiation potential for the generation of early cardiogenic progenitors, but they fail to transition into functional cardiomyocytes. In this work, by using a series of transcriptomic, cell and biochemical analyses, and the Qki-deficient mouse model, we demonstrate that QKI is indispensable to cardiac sarcomerogenesis and cardiac function through its regulation of alternative splicing in genes involved in Z-disc formation and contractile physiology, suggesting that QKI is associated with the pathogenesis of certain forms of cardiomyopathies.

RNA binding proteins (RBPs) have a broad biological and physiological function and are critical in regulating pre-mRNA posttranscriptional processing, intracellular migration, and mRNA stability. QKI, also known as Quaking, is a member of the signal transduction and activation of RNA (STAR) family, which also belongs to the heterogeneous nuclear ribonucleoprotein K- (hnRNP K-) homology domain protein family. There are three major alternatively spliced isoforms, QKI-5, QKI-6, and QKI-7, differing in carboxy-terminal domains. They share a common RNA binding property, but each isoform can regulate pre-mRNA splicing, transportation or stability differently in a unique cell type-specific manner. Previously, QKI has been known for its important role in contributing to neurological disorders. A series of recent work has further demonstrated that QKI has important roles in much broader biological systems, such as cardiovascular development, monocyte to macrophage differentiation, bone metabolism, and cancer progression. In this mini-review, we will focus on discussing the emerging roles of QKI in regulating cardiac and vascular development and function and its potential link to cardiovascular pathophysiology.

Immunotherapies for cancers may cause severe and life-threatening cardiotoxicities. The underlying mechanisms are complex and largely elusive. Currently, there are several ongoing clinical trials based on the use of activated invariant natural killer T (iNKT) cells. The potential cardiotoxicity commonly associated with this particular immunotherapy has yet been carefully evaluated. The present study aims to determine the effect of activated iNKT cells on normal and β-adrenergic agonist (isoproterenol, ISO)-stimulated hearts. Mice were treated with iNKT stimulants, α-galactosylceramide (αGC) or its analog OCH, respectively, to determine their effect on ISO-induced cardiac injury. We showed that administration of αGC (activating both T helper type 1 (Th1)- and T helper type 2 (Th2)-liked iNKT cells) significantly accelerated the progressive cardiac injury, leading to enhanced cardiac hypertrophy and cardiac fibrosis with prominent increases in collagen deposition and TGF-β1, IL-6, and alpha smooth muscle actin expression. In contrast to αGC, OCH (mainly activating Th2-liked iNKT cells) significantly attenuated the progression of cardiac injury and cardiac inflammation induced by repeated infusion of ISO. Flow cytometry analysis revealed that αGC promoted inflammatory macrophage infiltration in the heart, while OCH was able to restrain the infiltration. In vitro coculture of αGC- or OCH-pretreated primary peritoneal macrophages with primary cardiac fibroblasts confirmed the profibrotic effect of αGC and the antifibrotic effect of OCH. Our results demonstrate that activating both Th1- and Th2-liked iNKT cells is cardiotoxic, while activating Th2-liked iNKT cells is likely cardiac protective, which has implied key differences among subpopulations of iNKT cells in their response to cardiac pathological stimulation.

Bone morphogenetic protein 10 (BMP10) is a cardiac peptide growth factor belonging to the transforming growth factor β superfamily that critically controls cardiovascular development, growth, and maturation. It has been shown that BMP10 elicits its intracellular signaling through a receptor complex of activin receptor-like kinase 1 with morphogenetic protein receptor type II or activin receptor type 2A. Previously, we generated and characterized a transgenic mouse line expressing BMP10 from the α-myosin heavy chain gene promoter and found that these mice have normal cardiac hypertrophic responses to both physiological and pathological stimuli. In this study, we report that these transgenic mice exhibit significantly reduced levels of cardiomyocyte apoptosis and cardiac fibrosis in response to a prolonged administration of the β-adrenoreceptor agonist isoproterenol. We further confirmed this cardioprotective function with a newly generated conditional Bmp10 transgenic mouse line, in which Bmp10 was activated in adult hearts by tamoxifen. Moreover, the intraperitoneal administration of recombinant human BMP10 was found to effectively protect hearts from injury, suggesting potential therapeutic utility of using BMP10 to prevent heart failure. Gene profiling and biochemical analyses indicated that BMP10 activates the SMAD-mediated canonical pathway and, unexpectedly, also the signal transducer and activator of transcription 3 (STAT3)-mediated signaling pathway both in vivo and in vitro Additional findings further supported the notion that BMP10's cardioprotective function likely is due to its dual activation of SMAD- and STAT3-regulated signaling pathways, promoting cardiomyocyte survival and suppressing cardiac fibrosis.

A new type of steel concrete composite beam with double-joist steel sections encased was introduced.Six large-scale specimens were designed and tested under four-point bending condition to study the performance indicators of normal section bending bearing capacity, deformation, ductility and ultimate curvature and ultimate bending moment.It was found that the double-steel reinforced concrete beam had high strength and well ductility.The cracking load and ultimate load of the beam were influenced by the steel ratio, rebar ratio, stirrup ratio, steel size, space, and shear studs.Based on the comparison of the test results and through correcting coefficient of concrete part and the whole part in two Chinese codes, the bending calculation formula of double-steel reinforced concrete beam was obtained.

BACKGROUND:Prenatal exposure to Lipopolysaccharide (LPS) produces hypertension in adult offspring rats. The present study was to explore the effects of prenatal inflammation on morphological and functional changes in the aorta from offspring rats and to further assess its susceptibility to cardiovascular diseases.METHODS AND RESULTS:Pregnant rats were treated intraperitoneally on gestation Days 8, 10 and 12 with saline, LPS (0.79 mg/kg), or pyrrolidine dithiocarbamate (PDTC, 100 mg/kg)+LPS, respectively. Aortic ring reactivity and histopathological alteration were analyzed in offspring at the age of 12 weeks. The detections of connexin (Cx) 37, Cx40, Cx43, and Cx45, including immunofluorescent patterns, protein levels and mRNA expression in the aorta, were performed as well. Furthermore, the expressions of Nuclear factor (NF)-κB (p65), IκBα, phospho-IκBα and IκBβ were determined. The results showed that prenatal LPS exposure leads to morphological abnormalities and impaired aortic reactivity in offspring. Prenatal LPS exposure also decreased the protein and mRNA expression of Cx37 in the aorta from offspring rats. NF-κB and phospho-IκBα levels were both increased, IκBα level, however, was decreased in the aorta of offspring from the maternal LPS exposure compared to the controls. Simultaneously, PDTC treatment markedly reversed the action of LPS.CONCLUSIONS:Decreased expression of Cx37 contributed to the aortic dysfunction of prenatal LPS exposure offspring, which should be associated with NF-κB activation.

Tumor-associated macrophages are a prominent component of lung cancer stroma and contribute to tumor progression. The protein V-set and Ig domain-containing 4 (VSIG4), a novel B7 family-related macrophage protein that has the capacity to inhibit T-cell activation, has a potential role in the development of lung cancer. In this study, 10 human non-small-cell lung cancer specimens were collected and immunohistochemically analyzed for VSIG4 expression. Results showed massive VSIG4+ cell infiltration throughout the samples. Immunofluorescent double staining showed that VSIG4 was present on CD68+ macrophages, but absent from CD3+ T cells, CD31+ endothelial cells, and CK-18+ epithelial cells. Moreover, VSIG4 was coexpressed on B7-H1+ and B7-H3+ cells in these tumor specimens. Transfection of the VSIG4 gene into 293FT cells demonstrated that the VSIG4 signal could inhibit cocultured CD4+ and CD8+ T-cell proliferation and cytokine (IL-2 and IFN-γ) production in vitro. Interestingly, in a murine tumor model induced by Lewis lung carcinoma cell line, we found that tumors grown in VSIG4-deficient (VSIG4−/−) mice were significantly smaller than those found in wild-type littermates. All of these results demonstrate that macrophage-associated VSIG4 is an activator that facilitates lung carcinoma development. Specific targeting of VSIG4 may prove to be a novel, efficacious strategy for the treatment of this carcinoma.

Based on a high-rise building in Guangdong Province and its large-span transfer structure,this paper put forward new constructional composite form with the double-steel reinforced concrete transfer( DSRCT) beam-column joint,and two low-cyclic reversed loading tests were conducted on the joints of two DSRCT beams under vertical and horizontal loads. The failure pattern,bending strength,stiffness,hysteretic characteristics,ductility,and energy dissipation capacity of the joints,and strain of reinforcements and shape steel at critical positions were all studied.Experimental results reveal: transfer beam with a built-in double steel web form a closed space constraints for concrete can improve the node region concrete shear capacity; hysteresis curves of double steel reinforced concrete transfer beam of are full; the deformation capacity,bearing capacity,stiffness,ductility and energy dissipation capacity are also improved; the end plate bolted connection for the converted column and double steel reinforced concrete beam can result in the development of plastic hinge at bottom of the converted column first,ensuring the mechanisms of‘strong beam and column and stronger joints' and‘strong transfer structure'in the seismic design.

BACKGROUND:Programmed death-1 (PD-1, Pdcd1)-deficient mice develop different types of autoimmune diseases depending on the mouse strain but its role in uterus development has not been reported.METHODS:In this study, the expression of PD-1 and its ligands, PD-L1 and PD-L2, in uterine tissues from aged WT mice in a 129svEv-Brd background was analyzed by immunohistochemistry and the uterine morphology between WT and PD-1-/- mice was compared by hematoxylin and eosin staining.RESULTS:The aged PD-1-/- female mice in a 129svEv-Brd rather than Balb/c background develop endometrial hyperplasia. H&E staining showed an increase in the number of glands, neovascularization and an extremely large luminal cavity in aged PD-1-/- uteri. Immunohistochemical assay showed that the expression of PD-1 was observed in glandular/luminal epithelium and cells infiltrating the stroma. Fluorescent double staining demonstrated that PD-1 expresses on CD68+ macrophages, CD3+ T cells, CD16+ monocytes, CD56+ NK cells and CK-18+ epithelial cells, respectively. Additionally, PD-1 co-expresses with vascular endothelial growth factor (VEGF), and PD-1 deficiency resulted in an accumulation of glandular/luminal epithelium derived VEGF, which accelerates the expression of the proliferation-associated protein, proliferating cell nuclear antigen (PCNA), and thus potentially lead to epithelial proliferation in aged PD-1-/- uteri.CONCLUSIONS:These findings showed that PD-1 deficiency augments luminal epithelial cell proliferation probably through induced VEGF secretion, suggesting PD-1 plays an important role in controlling the growth and differentiation of the uterine epithelium.VIRTUAL SLIDES:The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/5809067461223905.

TNF-α plays an essential role in the pathogenesis of fulminant virus hepatitis (FH) caused by infection with murine hepatitis virus strain-3 (MHV-3). However, the specific TNF-α receptors (TNFR) involved in this disease and how they mediate this effect are uncertain. Here, we showed that the expression of TNFR1 and TNFR2 in the liver and spleen was triggered by MHV-3. However, only TNFR1−/− mice were resistant to MHV-3 mediated FH, as displayed by a dramatic decrease in tissue necrosis and cell apoptosis in the infected spleens and livers from TNFR1−/− mice, as well as prolonged survival in these mice compared to wild type littermate controls. Mechanistically, TNFR1 deficiency directly impeded the serum and tissue levels of fibrinogen-like protein 2 (FGL2), a virus-induced procoagulant molecule that promotes cell apoptosis. Additionally, the expression of apoptosis-associated molecules, Fas and Fas ligand (FasL) in the infected organs from TNFR1−/− mice were also decreased. Moreover, the infiltration of neutrophils rather than Foxp3+ regulatory T cells, which produce proinflammatory factors and FGL2 directly, into the infected liver and spleen tissues was also decreased in TNFR1−/− mice. These combined results indicate that signaling through TNFR1 plays an essential role in the pathogenesis of FH caused by MHV-3 infection, and interruption of this signaling pathway could be useful for clinical therapy.

Background: Endothelial progenitor cells (EPCs) derived from the bone marrow (BM) play a key role in the homeostasis of vascular repair by enhanced reendothelialization. Panax notoginseng saponins (PNS), a highly valued traditional Chinese medicine, has been shown to reduce morbidity and mortality from coronary artery disease. The present research was designed to explore the contribution of progenitor cells to the progression of atherosclerotic plaques and the possible modulatory role of PNS in this process. Methods: PNS (60 or 120 mg/kg via intraperitoneal injection) was administered over 8 weeks in apolipoprotein E knockout mice on an atherogenic diet. The sizes and histochemical alteration of atherosclerotic lesions and numbers of EPCs in BM and peripheral blood were analyzed. The expression of chemokine stromal cell-derived factor 1α (SDF-1α) and its receptor, CXCR4, was monitored as well. Results: PNS significantly reduced the lesion area and intima-to-media ratio compared to vehicle treatment. PNS also augmented endothelialization and reduced the smooth muscle cell (SMCs) content of the lesions. The number of c-kit and sca-1 double-positive progenitor cells and flk-1 and sca-1 double-positive progenitor cells were significantly increased in the BM and the peripheral blood of the PNS-treated groups. PNS treatment increased the plasma levels of SDF-1α and SCF as well as the BM levels of matrix metalloproteinase-9 (MMP-9). Moreover, the mRNA levels of SDF-1α and protein levels of CXCR4 were both increased in the BM of mice treated with PNS, while SDF-1α expression decreased. Conclusion: PNS reduce the size of atherosclerotic plaques, and this effect appears to involve progenitor cell mobilization. SDF-1α-CXCR4 interactions and the possible modulatory role of PNS in this process may contribute to the increased progenitor cell mobilization.

Signals from the T cell immunoglobulin and mucin-domain (TIM)-containing molecules have been demonstrated to be involved in regulating the progress of carcinoma. However, the expression and anatomical distribution of TIMs in Langerhans cell sarcoma (LCS), which is a rare malignancy derived from dendritic cells of the epidermis, has yet to be determined. In this study, the expression of TIM-1, TIM-3 and TIM-4 in LCS samples were detected by immunohistochemistry. Our results showed that these three molecules were found in LCS sections. At the cellular level, these molecules were found on the cell membrane and in the cytoplasm. Immunofluorescence double-staining demonstrated that these TIMs were co-expressed with Langerin, a potential biomarker for detecting LCS. In addition, TIM-1 was also expressed on CD68(+) macrophages and CK-18(+) epithelial cells, while TIM-3 and TIM-4 were expressed on all cell types investigated, including CD3(+)T cells, CD68(+) macrophages, CD11c(+) dendritic cells, CD16(+) NK Cells, CD31(+) endothelial cells and CK-18(+) epithelial cells. Interestingly, TIMs were also co-expressed with some members of the B7 superfamily, including B7-H1, B7-H3 and B7-H4 on sarcoma cells. Our results clearly showed the characteristic expression and anatomical distribution of TIMs in LCS, and a clear understanding of their functional roles may further elucidate the pathogenesis of this carcinoma and potentially contribute to the development of novel immunotherapeutic strategies.