The interaction of transcription factors GATA4 and NKX2-5 and the effect of interaction-targeted small molecules on the heart

Heart failure, a progressive disease, is a consequence of various cardiac diseases. It is one of the leading causes of mortality in the world and a challenge for modern medicine. Although the primary prevention of cardiac diseases in developed countries has improved, the absolute number of heart failure patients is increasing. The current treatments relieve the symptoms and improve quality of life, but they only delay the progression of the disease. Cardiac hypertrophy initially develops as an adaptive response to reduce ventricular wall stress and maintain cardiac function. However, if hypertrophy is prolonged, it activates pathological remodelling processes, e.g. increased fibrosis and cell death, inadequate angiogenesis and upregulation of foetal gene expression. Sequence-specific DNA-binding proteins, transcription factors (TFs), regulate gene expression and coordinate cell specification. The TF GATA-binding protein 4 (GATA4) is involved in several remodelling processes. It mediates the hypertrophic growth of cardiomyocytes, induces angiogenesis, and regulates apoptosis and cell survival. GATA4 and another cardiac TF, NK2 homeobox 5 (NKX2-5), interact physically and synergistically upregulate gene expression of, e.g. the foetal genes. In addition, their physical co-operation is required for mechanical stretch-induced brain natriuretic peptide (BNP) activation and cardiomyocyte hypertrophy. Thus, GATA4-NKX2-5 interaction is an interesting novel drug target. The aim of this thesis was to (i) characterise the GATA4-NKX2-5 protein-protein interaction and determine the NKX2-5 binding site on the GATA4 protein, (ii) investigate small molecule binding to GATA4 and NKX2-5 and (iii) to study the effect of the small molecule compounds in experimental in vivo models of heart failure. Mutational studies revealed a specific region involving amino acid N272 on the surface of the GATA4 protein that mediates the interaction with NKX2-5. The inhibitor of GATA4-NKX2-5 interaction, 3i-1000, was demonstrated to bind GATA4. This compound presented anti-hypertrophic and cardioprotective effects in vivo. This work demonstrates that cardiac TFs can be targeted and their functions modulated with small molecule compounds. In addition, the results suggest that targeting protein-protein interactions of key TFs may present a novel strategy for treatment of heart failure

Early clinical features of new-onset refractory status epilepticus (NORSE) in adults

Peer reviewe

GATA4-targeted compound exhibits cardioprotective actions against doxorubicin-induced toxicity in vitro and in vivo : establishment of a chronic cardiotoxicity model using human iPSC-derived cardiomyocytes

Doxorubicin is a widely used anticancer drug that causes dose-related cardiotoxicity. The exact mechanisms of doxorubicin toxicity are still unclear, partly because most in vitro studies have evaluated the effects of short-term high-dose doxorubicin treatments. Here, we developed an in vitro model of long-term low-dose administration of doxorubicin utilizing human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Moreover, given that current strategies for prevention and management of doxorubicin-induced cardiotoxicity fail to prevent cancer patients developing heart failure, we also investigated whether the GATA4-targeted compound 3i-1000 has cardioprotective potential against doxorubicin toxicity both in vitro and in vivo. The final doxorubicin concentration used in the chronic toxicity model in vitro was chosen based on cell viability data evaluation. Exposure to doxorubicin at the concentrations of 1-3 mu M markedly reduced (60%) hiPSC-CM viability already within 48 h, while a 14-day treatment with 100 nM doxorubicin concentration induced only a modest 26% reduction in hiPCS-CM viability. Doxorubicin treatment also decreased DNA content in hiPSC-CMs. Interestingly, the compound 3i-1000 attenuated doxorubicin-induced increase in pro-B-type natriuretic peptide (proBNP) expression and caspase-3/7 activation in hiPSC-CMs. Moreover, treatment with 3i-1000 for 2 weeks (30 mg/kg/day, i.p.) inhibited doxorubicin cardiotoxicity by restoring left ventricular ejection fraction and fractional shortening in chronic in vivo rat model. In conclusion, the results demonstrate that long-term exposure of hiPSC-CMs can be utilized as an in vitro model of delayed doxorubicin-induced toxicity and provide in vitro and in vivo evidence that targeting GATA4 may be an effective strategy to counteract doxorubicin-induced cardiotoxicity.Peer reviewe

納税者の行為と納税者以外の者の行為――重加算税が課せられる行為と共同事業者の行為の対比を契機として――（一）

As companies have recently gotten more interested in utilizing the increasingly gathered data and realizing the potential of data analysis, the ability to upgrade data into value for business has been recognized as an advantage. Companies gain competitive advantage if they are able to benefit from the fleet data that is produced both in and outside the boundaries of the company. Benefits of fleet management are based on the possibility to have access to the massive amounts of asset data that can then be utilized e.g. to gain cost savings and to develop products and services. The ambition of the companies is to create value from fleet data but this requires that different actors in ecosystem are working together for a common goal – to get the most value out of fleet data for the ecosystem. In order that this could be possible, we need a framework to meet the requirements of the fleet life-cycle data utilization. This means that the different actors in the ecosystem need to understand their role in the fleet data refining process in order to promote the value creation from fleet data. The objective of this paper is to develop a framework for knowledge management in order to create value from fleet data in ecosystems. As a result, we present a conceptual framework which helps companies to develop their asset management practices related to the fleet of assets.Jako, że przedsiębiorstwa w ostatnim czasie zaczęły bardziej interesować się wykorzystaniem wzrastającej ilości danych i zdały sobie sprawę z potencjału analizy danych, uznano, że zdolność do przekształcenia danych w wartość dla przedsiębiorstwa jest dla niego korzystne. Przedsiębiorstwa zdobywają przewagę konkurencyjną, jeśli są w stanie wykorzystać dane floty, które są generowane zarówno wewnątrz, jak i na zewnątrz przedsiębiorstwa. Korzyści płynące z zarządzania flotą opierają się na możliwości uzyskania dostępu do ogromnych ilości danych o aktywach, które można następnie wykorzystać np. w celu uzyskania oszczędności a także rozwoju produktów i usług. Celem przedsiębiorstw jest tworzenie wartości z danych floty, ale wymaga to, aby różne podmioty w ekosystemie współpracowały ze sobą we wspólnym celu – aby uzyskać najwyższą wartość z danych floty dla ekosystemu. Aby było to możliwe, potrzebne są ramy, aby spełnić wymagania dotyczące wykorzystania danych o cyklu życia floty. Oznacza to, że różne podmioty w ekosystemie muszą zrozumieć swoją rolę w procesie rafinacji danych floty w promowaniu tworzenia wartości z danych floty. Celem niniejszego artykułu jest opracowanie ram zarządzania wiedzą w tworzeniu wartości z danych floty w ekosystemach. Jako wynik przedstawiono koncepcyjne podstawy, które pomagają przedsiębiorstwom rozwijać praktyki zarządzania aktywami związane z flotą aktywów

GATA-targeted compounds modulate cardiac subtype cell differentiation in dual reporter stem cell line

BackgroundPharmacological modulation of cell fate decisions and developmental gene regulatory networks holds promise for the treatment of heart failure. Compounds that target tissue-specific transcription factors could overcome non-specific effects of small molecules and lead to the regeneration of heart muscle following myocardial infarction. Due to cellular heterogeneity in the heart, the activation of gene programs representing specific atrial and ventricular cardiomyocyte subtypes would be highly desirable. Chemical compounds that modulate atrial and ventricular cell fate could be used to improve subtype-specific differentiation of endogenous or exogenously delivered progenitor cells in order to promote cardiac regeneration.MethodsTranscription factor GATA4-targeted compounds that have previously shown in vivo efficacy in cardiac injury models were tested for stage-specific activation of atrial and ventricular reporter genes in differentiating pluripotent stem cells using a dual reporter assay. Chemically induced gene expression changes were characterized by qRT-PCR, global run-on sequencing (GRO-seq) and immunoblotting, and the network of cooperative proteins of GATA4 and NKX2-5 were further explored by the examination of the GATA4 and NKX2-5 interactome by BioID. Reporter gene assays were conducted to examine combinatorial effects of GATA-targeted compounds and bromodomain and extraterminal domain (BET) inhibition on chamber-specific gene expression.ResultsGATA4-targeted compounds 3i-1000 and 3i-1103 were identified as differential modulators of atrial and ventricular gene expression. More detailed structure-function analysis revealed a distinct subclass of GATA4/NKX2-5 inhibitory compounds with an acetyl lysine-like domain that contributed to ventricular cells (%Myl2-eGFP+). Additionally, BioID analysis indicated broad interaction between GATA4 and BET family of proteins, such as BRD4. This indicated the involvement of epigenetic modulators in the regulation of GATA-dependent transcription. In this line, reporter gene assays with combinatorial treatment of 3i-1000 and the BET bromodomain inhibitor (+)-JQ1 demonstrated the cooperative role of GATA4 and BRD4 in the modulation of chamber-specific cardiac gene expression.ConclusionsCollectively, these results indicate the potential for therapeutic alteration of cell fate decisions and pathological gene regulatory networks by GATA4-targeted compounds modulating chamber-specific transcriptional programs in multipotent cardiac progenitor cells and cardiomyocytes. The compound scaffolds described within this study could be used to develop regenerative strategies for myocardial regeneration.Peer reviewe

GSK3β Serine 389 Phosphorylation Modulates Cardiomyocyte Hypertrophy and Ischemic Injury

Prior studies show that glycogen synthase kinase 3β (GSK3β) contributes to cardiac ischemic injury and cardiac hypertrophy. GSK3β is constitutionally active and phosphorylation of GSK3β at serine 9 (S9) inactivates the kinase and promotes cellular growth. GSK3β is also phosphorylated at serine 389 (S389), but the significance of this phosphorylation in the heart is not known. We analyzed GSK3β S389 phosphorylation in diseased hearts and utilized overexpression of GSK3β carrying ser→ala mutations at S9 (S9A) and S389 (S389A) to study the biological function of constitutively active GSK3β in primary cardiomyocytes. We found that phosphorylation of GSK3β at S389 was increased in left ventricular samples from patients with dilated cardiomyopathy and ischemic cardiomyopathy, and in hearts of mice subjected to thoracic aortic constriction. Overexpression of either GSK3β S9A or S389A reduced the viability of cardiomyocytes subjected to hypoxia–reoxygenation. Overexpression of double GSK3β mutant (S9A/S389A) further reduced cardiomyocyte viability. Determination of protein synthesis showed that overexpression of GSK3β S389A or GSK3β S9A/S389A increased both basal and agonist-induced cardiomyocyte growth. Mechanistically, GSK3β S389A mutation was associated with activation of mTOR complex 1 signaling. In conclusion, our data suggest that phosphorylation of GSK3β at S389 enhances cardiomyocyte survival and protects from cardiomyocyte hypertrophy

GSK3β Serine 389 Phosphorylation Modulates Cardiomyocyte Hypertrophy and Ischemic Injury

Prior studies show that glycogen synthase kinase 3β (GSK3β) contributes to cardiac ischemic injury and cardiac hypertrophy. GSK3β is constitutionally active and phosphorylation of GSK3β at serine 9 (S9) inactivates the kinase and promotes cellular growth. GSK3β is also phosphorylated at serine 389 (S389), but the significance of this phosphorylation in the heart is not known. We analyzed GSK3β S389 phosphorylation in diseased hearts and utilized overexpression of GSK3β carrying ser→ala mutations at S9 (S9A) and S389 (S389A) to study the biological function of constitutively active GSK3β in primary cardiomyocytes. We found that phosphorylation of GSK3β at S389 was increased in left ventricular samples from patients with dilated cardiomyopathy and ischemic cardiomyopathy, and in hearts of mice subjected to thoracic aortic constriction. Overexpression of either GSK3β S9A or S389A reduced the viability of cardiomyocytes subjected to hypoxia–reoxygenation. Overexpression of double GSK3β mutant (S9A/S389A) further reduced cardiomyocyte viability. Determination of protein synthesis showed that overexpression of GSK3β S389A or GSK3β S9A/S389A increased both basal and agonist-induced cardiomyocyte growth. Mechanistically, GSK3β S389A mutation was associated with activation of mTOR complex 1 signaling. In conclusion, our data suggest that phosphorylation of GSK3β at S389 enhances cardiomyocyte survival and protects from cardiomyocyte hypertrophy

Ammattikorkeakouluopettajien monialainen sosiaali- ja terveydenhuollon tiedonhallinnan osaaminen

The quick digital transformation (DT) in society has affected the development of service in health and social services strongly during the last five years. The digitalization has resulted in a change in the paradigm, which has required a strong reaction from education to respond to the needs of competence in working life. The students who graduate from a university of applied sciences must be provided with sufficient readiness to operate in working life and to cope with change. The precondition is that the educators of health and social care, business, service design and IT keep up with the development of digitalization. The purpose of this study was to evaluate the informatics competences of the educators at the universities of applied sciences with an emphasis on the operational environments of health and social services. The survey is a part of the SotePeda 24/7 project. The survey was sent to Finnish universities of applied sciences (22) participating in the project in April- May 2019. The survey form was based on national and international evidence-based information. The form included 12 fields of competence and their contents. 172 answers were obtained after three reminders. A factor analysis was conducted utilizing the maximum likelihood method and the result was 18 factors. The largest load factor was named as the information management and digitalization competence factor. This article discusses the informatics variables loaded for this factor in the survey. The correspondence and discriminant analyses conducted in the study showed the competences vary in different education fields. The biggest differences in informatics competence were found in the social services education field, whereas the competences of the educators of the health care education were good and extremely good more evenly. In other fields participating in the study there were big differences inside single education fields regarding the informatics competences of health and social services.Nopea yhteiskunnan digitaalinen muutos (Digital Transformation, DT) on vaikuttanut viimeisten viiden vuoden aikana vahvasti sosiaali- ja terveydenhuollon palvelukehitykseen. Digitalisaatio on synnyttänyt paradigman muutoksen, mihin koulutuksen on pitänyt vahvasti reagoida vastatakseen vaadittaviin työelämän osaamistarpeisiin. Ammattikorkeakoulusta valmistuvalle ammattilaiselle on koulutuksessa taattava riittävät valmiudet toimia työelämässä ja selviytyä muutoksesta. Edellytyksenä on, että sosiaali- ja terveysalan, liiketalouden, palvelumuotoilualan ja IT-alan opettajat hallitsevat digitalisaation kehityksen. Tutkimuksen tarkoituksena oli arvioida ammattikorkeakouluopettajien sosiaali- ja terveydenhuollon digitaalisessa toimintaympäristössä tarvittavaa tiedonhallinnan osaamista. Tutkimus on osa SotePeda 24/7 -hanketta. Hankkeessa mukana oleville Suomen ammattikorkeakouluille (22) lähetettiin huhti-toukokuussa 2019 kyselylomake, joka pohjautui kansalliseen ja kansainväliseen näyttöön perustuvaan tietoon. Lomakkeessa oli 12 osaamisaluetta sisältöineen. Vastauksia saatiin kolmen muistutuksen jälkeen 172. Aineisto analysoitiin faktorianalyysillä hyödyntäen maximum likelihood menetelmää, ja tuloksena saatiin 18 faktoria. Suurimman latauksen saanut faktori nimettiin tiedonhallinnan ja digitaalisuuden osaamisen faktoriksi. Tässä artikkelissa tarkastellaan kyselyssä tälle faktorille latautuneita tiedonhallinnan muuttujia. Korrespondenssianalyysit ja tarkentavat erotteluanalyysit osoittivat osaamisen eroavan eri koulutusaloilla. Sosiaalialalla tiedonhallinnan osaamisessa oli koulutusaloista isommat erot kuin terveysalan opettajilla, joiden osaaminen oli tasaisemmin hyvää ja erittäin hyvää. Sosiaali- ja terveydenhuollon tiedonhallinnan osaamisessa oli muilla tutkimukseen osallistuvilla aloilla isoja eroja oman koulutusalan sisällä