Journal of molecular and cellular cardiology plus最新文献_第4页

Generation and evaluation of a novel PI3K-targeting gene therapy in the failing mouse heart and healthy sheep heart 一种新的pi3k靶向基因治疗衰竭小鼠心脏和健康绵羊心脏的产生和评估

IF 2.2

Journal of molecular and cellular cardiology plus

Pub Date : 2025-07-31 DOI: 10.1016/j.jmccpl.2025.100478

Sebastian Bass-Stringer , Daniel G. Donner , Clive N. May , Aya Matsumoto , Emma I. Masterman , Aascha A. D'Elia , Yi Ching Chen , Helen Kiriazis , Jieting Luo , Roger Chooi , Clara Liu Chung Ming , Paul Gregorevic , Colleen J. Thomas , Bianca C. Bernardo , Kate L. Weeks , Julie R. McMullen

Heart failure (HF) remains a clinical challenge with cardiac dysfunction typically progressing even with treatment, and heart transplants only available to small numbers. We previously identified phosphoinositide 3-kinase (PI3K, p110α) as a master regulator of exercise-induced cardioprotection, and showed that gene therapy, incorporating a constitutively active form of PI3K (caPI3K) improved function of the failing mouse heart. However, this approach was not cardiac-specific and the gene therapy was challenging to manufacture. The aim of this study was to develop new PI3K-based gene therapies with more optimal properties for clinical translation. We generated and assessed adeno-associated viruses (AAV6) encoding various PI3K constructs, with different enhancers, promoters and transgene components in healthy adult male mice. The most promising AAV construct based on AAV expression, cardiac-specificity, and ease of manufacture contained a cardiac troponin T (cTnT) promoter together with a small region of the regulatory subunit of PI3K (iSH2), and an intron from the β-globin gene which enhances transcription (IVS2). This AAV (1 × 10¹², 2 × 10¹² vg) was administered to mice with myocardial ischemia/reperfusion injury (I/R: 1 h ischemia with reperfusion; AAV delivered 24 h post-I/R). Direct cardiac injections of PI3K-based AAVs were also performed in healthy adult female sheep. I/R mouse hearts treated with the AAV6-cTnT-IVS2-iSH2 displayed increased phosphorylation of Akt, but no improvement in cardiac function or structure was observed. AAV6-cTnT-IVS2-iSH2 successfully transduced healthy sheep hearts which increased endogenous PI3K catalytic activity. Further testing/optimization of the AAV (time of delivery and/or duration) will be required to assess the therapeutic potential of this approach.

心力衰竭（HF）仍然是一个临床挑战，即使治疗，心脏功能障碍通常也会恶化，心脏移植只适用于少数人。我们之前确定了磷酸肌肽3-激酶（PI3K, p110α）是运动诱导的心脏保护的主要调节因子，并表明基因治疗，包括组成活性形式的PI3K （caPI3K）改善了衰竭小鼠的心脏功能。然而，这种方法不是针对心脏的，而且这种基因疗法的制造具有挑战性。本研究的目的是开发新的基于pi3的基因疗法，具有更理想的临床转化特性。我们在健康成年雄性小鼠中生成并评估了编码多种PI3K构建体、具有不同增强子、启动子和转基因成分的腺相关病毒（AAV6）。基于AAV表达、心脏特异性和易于制造的最有希望的AAV构建物包含一个心脏肌钙蛋白T （cTnT）启动子、PI3K调控亚基的一个小区域（iSH2）和一个来自β-珠蛋白基因的内含子，该内含子可促进转录（IVS2）。将AAV （1 × 1012,2 × 1012vg）给予心肌缺血/再灌注损伤小鼠(I/R：缺血再灌注1 h；AAV在i /R后24小时交付)。在健康成年母羊中也进行了基于pi3的aav直接心脏注射。AAV6-cTnT-IVS2-iSH2处理的I/R小鼠心脏显示Akt磷酸化增加，但未观察到心脏功能或结构的改善。AAV6-cTnT-IVS2-iSH2成功转导了健康绵羊心脏，增加了内源性PI3K的催化活性。需要进一步测试/优化AAV（给药时间和/或持续时间）来评估该方法的治疗潜力。

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引用次数: 0

Current status and challenges of multi-omics research using animal models of atherosclerosis 动脉粥样硬化动物模型多组学研究现状与挑战

Journal of molecular and cellular cardiology plus

Pub Date : 2025-07-10 DOI: 10.1016/j.jmccpl.2025.100476

Tijana Mitić , Adriana Georgescu , Nicoleta Alexandru-Moise , Michael J. Davies , Cecile Vindis , Susana Novella , Eva Gerdts , Georgios Kararigas , Stephanie Bezzina Wettinger , Melissa M. Formosa , Brenda R. Kwak , Filippo Molica , Nuria Amigo , Andrea Caporali , Fernando de la Cuesta , Ignacio Fernando Hall , Angeliki Chroni , Fabio Martelli , Johannes A. Schmid , Paolo Magni , Dimitris Kardassis

Atherosclerosis is an underlying cause of cardiovascular diseases (CVD) which account for most deaths worldwide. Use of diverse preclinical models of atherosclerosis has been implemental in understanding the underlying mechanisms, the implicated cell types, the genes and the molecules at play in the onset and progression of atherosclerotic plaques. Although significant research advancements have been made, further research is necessary to delve into factors influencing plaque types, site preference within the vasculature, interactions with adjacent tissues (liver, pancreas and perivascular adipose tissue), inflammation and sex-based disparities, among others. The conventional low throughput methodologies which concentrate on individual cells, genes or metabolites are inadequate to tackle the complex and heterogeneous nature of atherosclerosis. With recent advancement in multi-omics and bioinformatics, research approaches have illuminated a clearer understanding of atherosclerosis. Consequently, these advancements pave the path to design novel therapeutics to complement currently approved lipid-lowering and other effective treatments. In this article, we summarize and critically evaluate the findings derived from recent high throughput single- or multi-omic studies conducted in animal models of atherosclerosis. We also delve into the challenges associated with using experimental animals to model human atherosclerosis and contemplate the essential enhancements needed to better mimic human conditions. We further discuss the requirement of establishing a structured multi-omic database for atherosclerosis research, enabling broader access and utilisation within the scientific community.

动脉粥样硬化是心血管疾病（CVD）的潜在原因，心血管疾病占全世界死亡人数最多。多种临床前动脉粥样硬化模型的应用有助于理解动脉粥样硬化斑块发生和发展的潜在机制、相关细胞类型、基因和分子。尽管已经取得了重大的研究进展，但需要进一步的研究来深入研究影响斑块类型、血管内的部位偏好、与邻近组织（肝脏、胰腺和血管周围脂肪组织）的相互作用、炎症和性别差异等因素。传统的专注于单个细胞、基因或代谢物的低通量方法不足以解决动脉粥样硬化的复杂性和异质性。随着近年来多组学和生物信息学的进展，研究方法已经阐明了动脉粥样硬化的更清晰的认识。因此，这些进步为设计新的治疗方法铺平了道路，以补充目前批准的降脂和其他有效的治疗方法。在本文中，我们总结并批判性地评估了最近在动脉粥样硬化动物模型中进行的高通量单组或多组研究的结果。我们还深入研究了与使用实验动物来模拟人类动脉粥样硬化相关的挑战，并考虑了更好地模拟人类条件所需的基本增强。我们进一步讨论了为动脉粥样硬化研究建立一个结构化的多组学数据库的需求，使其在科学界得到更广泛的访问和利用。

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引用次数: 0

Cardiology's best friend: Using naturally occurring disease in dogs to understand heart disease in humans 心脏病学最好的朋友：用狗身上自然发生的疾病来了解人类的心脏病

Journal of molecular and cellular cardiology plus

Pub Date : 2025-07-04 DOI: 10.1016/j.jmccpl.2025.100474

W. Glen Pyle

Heart diseases are a leading cause of death globally. Laboratory and preclinical animal models of disease have been critical in advancing our understanding of the mechanisms of pathology, creating diagnostic tools, and developing therapeutic interventions. However, fundamental biological dissimilarities between humans and rodents limits their usefulness in research, and the induction of disease in an otherwise healthy animal creates unrealistic conditions under which diseases are typically studied. Dogs are at high risk of acquiring and dying from several naturally occurring heart disorders that also affect people. The spontaneous nature of these conditions, along with highly similar cardiovascular systems, offers unique opportunities to investigate cardiovascular disease in a more relevant model for humans. This review focuses on three common cardiac conditions that impact humans and dogs: dilated cardiomyopathy, arrhythmogenic right ventricular cardiomyopathy, and mitral valve disease – comparing mechanisms of disease, diagnostics, and treatments, to identify strengths and present limitations of their utility. It is noted that the benefits of this research are bidirectional, with the potential to translate knowledge and clinical tools used in veterinary medicine to human patients, and vice versa.

心脏病是全球死亡的主要原因。疾病的实验室和临床前动物模型对于促进我们对病理机制的理解、创建诊断工具和开发治疗干预措施至关重要。然而，人类和啮齿类动物之间基本的生物学差异限制了它们在研究中的用处，而且在健康的动物身上诱发疾病创造了不现实的条件，而疾病通常是在这种条件下进行研究的。狗有很高的风险患上和死于几种自然发生的心脏疾病，这些疾病也会影响到人类。这些疾病的自发性，以及高度相似的心血管系统，为在更相关的人类模型中研究心血管疾病提供了独特的机会。本文综述了影响人类和狗的三种常见心脏疾病：扩张型心肌病、致心律失常的右室心肌病和二尖瓣疾病——比较疾病的机制、诊断和治疗，以确定它们的优势和局限性。值得注意的是，这项研究的好处是双向的，有可能将兽医中使用的知识和临床工具转化为人类患者，反之亦然。

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引用次数: 0

Myocardial damage post short-term self-administration cocaine usage in rat 大鼠短期自用可卡因后心肌损伤

Journal of molecular and cellular cardiology plus

Pub Date : 2025-07-01 DOI: 10.1016/j.jmccpl.2025.100475

Muhammad Zubair Saleem , Tian Liu , Ningjing Song , Jayme McReynolds , Chen Gao

Cocaine abuse remains a significant risk with profound adverse impact on cardiovascular health. While long-term cocaine addiction-induced cardiotoxicity is well-documented, the underlying mechanism and the molecular effects of short-term recreational usage of cocaine on the heart have not been well studied. We established a short-term cocaine exposure rat model through self-administration, mimicking real-world recreational cocaine usage. Our results indicate that even such short-term cocaine usage induces deleterious effect on the heart including pathological remodeling and transcriptome reprogramming associated with major metabolic and contractile processes. This study sheds important insight on the molecular mechanisms of short-term exposure of cocaine-induced cardiovascular damage.

可卡因滥用仍然是对心血管健康产生严重不利影响的重大风险。虽然长期可卡因成瘾引起的心脏毒性已被充分证明，但短期娱乐性可卡因使用对心脏的潜在机制和分子效应尚未得到很好的研究。我们通过自我给药建立了一个短期可卡因暴露大鼠模型，模仿真实世界的娱乐性可卡因使用。我们的研究结果表明，即使是这样的短期可卡因使用也会对心脏产生有害影响，包括与主要代谢和收缩过程相关的病理性重塑和转录组重编程。这项研究揭示了短期暴露于可卡因引起的心血管损伤的分子机制。

引用次数: 0

MitoQ Protects Against Oxidative Stress-Induced Mitochondrial Dysregulation in Human Cardiomyocytes MitoQ保护心肌细胞免受氧化应激诱导的线粒体失调

Journal of molecular and cellular cardiology plus

Pub Date : 2025-06-26 DOI: 10.1016/j.jmccpl.2025.100469

Alex M. Parker , Jarmon G. Lees , Mitchel Tate , Ren J. Phang , Anida Velagic , Minh Deo , Tayla Bishop , Thomas Krieg , Michael P. Murphy , Shiang Y. Lim , Miles J. De Blasio , Rebecca H. Ritchie

The overproduction of reactive oxygen species (ROS) and mitochondrial dysregulation are regarded as key mechanisms in the progression of cardiac remodelling in cardiometabolic diseases including heart failure. Conventional treatments are often ineffective as they do not specifically target the underlying pathological mechanisms. Mitoquinone mesylate (MitoQ), a mitochondrial-targeted antioxidant has been reported to be protective against vascular dysfunction in hypertension, diabetic kidney disease and alcohol-induced liver damage. However, the cardioprotective potential of MitoQ to limit oxidative stress-induced mitochondrial remodelling in cardiomyocytes has not been fully resolved. We sought to investigate the effect of MitoQ and its mitochondrial-targeting moiety dodecyl-triphenylphosphonium (dTPP) on hydrogen peroxide-induced overproduction of ROS, mitochondrial dysregulation and cell death in H9C2 rat cardiomyoblasts (H9C2-rCM) and human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM). Cardiomyocytes were exposed to acute or chronic treatment (5–60 min or 48 h) of vehicle control (0.0001 % Ultrapure Milli-Q water), hydrogen peroxide (100 μM) ± MitoQ (1 μM) or dTPP (1 μM) control. Hydrogen peroxide-induced overproduction of ROS, extracellular superoxide, mitochondrial ROS, mitochondrial hyperpolarisation and cell death were significantly blunted by MitoQ, but not dTPP, suggesting that the coenzyme Q₁₀ moiety of MitoQ is protective under these conditions. Interestingly, both MitoQ and dTPP exhibited a pro-mitochondrial fusion effect by preserving mitochondrial network and reducing mitochondrial fragmentation in oxidative stress conditions. Overall, our findings confirm the cytoprotective potential of MitoQ to limit oxidative stress-induced adverse mitochondrial remodelling and dysregulation that is clinically observed in cardiometabolic-induced cardiac dysfunction in the failing heart.

活性氧（ROS）的过量产生和线粒体失调被认为是心脏代谢疾病（包括心力衰竭）中心脏重构进展的关键机制。常规治疗往往是无效的，因为他们没有专门针对潜在的病理机制。甲磺酸米托醌（Mitoquinone mesylate, MitoQ）是一种线粒体靶向抗氧化剂，据报道可预防高血压、糖尿病肾病和酒精性肝损伤的血管功能障碍。然而，MitoQ限制心肌细胞氧化应激诱导的线粒体重构的心脏保护潜力尚未完全解决。我们试图研究MitoQ及其线粒体靶向片段十二烷基三苯磷（dTPP）对H9C2大鼠成心肌细胞（H9C2- rcm）和人诱导多能干细胞源性心肌细胞（hiPSC-CM）过氧化氢诱导的ROS过量产生、线粒体失调和细胞死亡的影响。心肌细胞暴露于急性或慢性对照（5-60分钟或48小时）的对照剂（0.0001 %超纯milliq水）、过氧化氢（100 μM）±MitoQ （1 μM）或dTPP （1 μM）。过氧化氢诱导的ROS过量产生、细胞外超氧化物、线粒体ROS、线粒体超极化和细胞死亡被MitoQ显著减弱，但dTPP没有，这表明MitoQ的辅酶Q10片段在这些条件下具有保护作用。有趣的是，在氧化应激条件下，MitoQ和dTPP都通过保留线粒体网络和减少线粒体碎片化而表现出促进线粒体融合的作用。总的来说，我们的研究结果证实了MitoQ的细胞保护潜力，以限制氧化应激诱导的不良线粒体重构和失调，这在临床上观察到心脏衰竭时心脏代谢诱导的心功能障碍。

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引用次数: 0

Semaphorin 3F is elevated in serum of heart failure patients and inhibits cardiac angiogenesis via the VEGF/Akt/eNOS pathway Semaphorin 3F在心力衰竭患者的血清中升高，并通过VEGF/Akt/eNOS途径抑制心脏血管生成

Journal of molecular and cellular cardiology plus

Pub Date : 2025-06-25 DOI: 10.1016/j.jmccpl.2025.100470

Diana Petrova , Miki Weberbauer , Stephanie Reichert , Stephanie Scheid , Jennifer Esser , Katrin Fink , Daniel Duerschmied , Martin Moser , Thomas Helbing

Left ventricular (LV) remodeling in heart failure (HF) is associated with vascular rarefaction and impaired angiogenesis. The inhibition of vascular endothelial growth factor (VEGF)-mediated angiogenesis is a key feature in the pathophysiology of HF. Semaphorin (Sema) 3F is a known inhibitor of VEGF signaling, but its role in HF remains to be elucidated.

Serum Sema3F levels were measured in HF patients (n = 70) by ELISA and were compared to those in patients with coronary artery disease (CAD, n = 26). Sema3F levels were significantly increased in HF patients. Sema3F RNA and protein expression were upregulated by hypoxia in cardiac endothelial cells (HCECs) as demonstrated by quantitative RT-PCR and Western blotting (WB). In Matrigel® sprouting assays, endothelial cell sprouting and branching were decreased in response to HF patient serum, suggesting that HF serum contains anti-angiogenic factors. Recombinant human Sema3F attenuated VEGF-mediated angiogenesis in Matrigel® sprouting, spheroid sprouting and aortic ring assays. Vice versa, siRNA-based Sema3F knockdown promoted angiogenesis. In zebrafish, morpholino-based Sema3F knockdown led to increased mortality and induced a vascular phenotype. Mechanistically, Sema3F inhibited VEGF-induced Akt and eNOS phosphorylation in endothelial cells, and Sema3F knockdown increased phosphorylation of Akt and eNOS.

Sema3F is elevated in serum of HF patients and has anti-angiogenic properties in cardiac angiogenesis through inhibition of the VEGF/Akt/eNOS pathway. Thus, targeting Sema3F could present a therapeutic approach to advanced HF in the future.

心力衰竭（HF）左心室（LV）重构与血管稀疏和血管生成受损有关。抑制血管内皮生长因子（VEGF）介导的血管生成是HF病理生理的一个关键特征。信号素（Sema） 3F是一种已知的VEGF信号抑制剂，但其在HF中的作用仍有待阐明。用ELISA法测定了HF患者（n = 70）的血清Sema3F水平，并与冠心病患者（n = 26）进行了比较。心衰患者的Sema3F水平显著升高。定量RT-PCR和Western blotting （WB）结果显示，缺氧导致心脏内皮细胞（HCECs）中Sema3F RNA和蛋白表达上调。在Matrigel®发芽试验中，内皮细胞发芽和分支减少，这表明HF患者血清含有抗血管生成因子。重组人Sema3F在Matrigel®发芽、球形发芽和主动脉环试验中减弱vegf介导的血管生成。反之亦然，基于sirna的Sema3F敲低促进血管生成。在斑马鱼中，基于morpholino的Sema3F敲低导致死亡率增加并诱导血管表型。在机制上，Sema3F抑制vegf诱导的内皮细胞中Akt和eNOS的磷酸化，Sema3F敲低增加Akt和eNOS的磷酸化。Sema3F在HF患者血清中升高，并通过抑制VEGF/Akt/eNOS通路在心脏血管生成中具有抗血管生成特性。因此，靶向Sema3F可能是未来晚期心衰的一种治疗方法。

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引用次数: 0

Combined genome and transcriptome analysis identifies molecular signatures of aortic disease in patients with Marfan syndrome 联合基因组和转录组分析确定马凡氏综合征患者主动脉疾病的分子特征

Journal of molecular and cellular cardiology plus

Pub Date : 2025-06-19 DOI: 10.1016/j.jmccpl.2025.100467

Katherine B. Stanley , Alexa V. Mederos , Ethan H. Barksdale , Joel S. Corvera , Joshua L. Davis , Fang Fang , Hongyu Gao , Courtney E. Vujakovich , Yunlong Liu , Stephanie M. Ware , Benjamin J. Landis

Introduction

Transcriptional dysregulation in patients with Marfan syndrome (MFS) is complex and not well-defined. There are likely patient-specific and general mechanisms in the aortic pathology. In this study, we combine genome and transcriptome data from patients with MFS to determine the transcriptional impacts of disease-causing variants in FBN1.

Methods

Prospectively enrolled participants provided blood and aortic tissue samples. Smooth muscle cells (SMCs) were cultured directly from the proximal aortic tissues of MFS cases undergoing aortic root replacement and controls during heart transplant. Genome sequencing (GS) analysis was combined with mRNA-sequencing (mRNA-seq) and single-cell gene expression profiling of SMCs. Findings in SMC culture analysis were further investigated in primary frozen aortic tissues.

Results

Automatic annotation of single-cell expression profiles classified 99% of cultured cells as SMCs. All disease-causing FBN1 variants were detected in both GS and SMC mRNA-seq reads. These included missense single nucleotide variants (SNVs), a whole-exon deletion, and a predicted stopgain SNV. Gene and allelic expression abnormalities in FBN1 were identified. Broadly, genes that were dysregulated in MFS were enriched for glycerophospholipid metabolism, immune, potassium channel, and extracellular matrix processes. Single-cell clustering analysis identified subtypes of SMCs. Some genes were differentially expressed in MFS across multiple SMC subtypes (e.g. TRPV2), whereas others were significant within specific SMC states (e.g. TGFB2 in SMCs expressing inflammatory markers).

Conclusions

mRNA-seq analysis of SMCs accurately identified FBN1 variants. General and patient-specific effects on allelic and gene expression were identified. Metabolism of glycerophospholipids may be dysregulated in aortic SMCs in MFS. Identifying pathogenic features with transcriptome analysis may guide novel diagnostic and therapeutic strategies.

马凡氏综合征（MFS）患者的转录失调是复杂且不明确的。主动脉病变可能有患者特异性和一般机制。在这项研究中，我们结合来自MFS患者的基因组和转录组数据来确定FBN1致病变异的转录影响。方法前瞻性招募的参与者提供血液和主动脉组织样本。平滑肌细胞（SMCs）是直接从心脏移植过程中行主动脉根置换术的MFS患者和对照组的主动脉近端组织中培养的。将基因组测序（GS）、mrna测序（mRNA-seq）和SMCs单细胞基因表达谱相结合进行分析。在原发冷冻主动脉组织中进一步研究SMC培养分析结果。结果单细胞表达谱自动注释将99%的培养细胞分类为SMCs。在GS和SMC的mRNA-seq读数中均检测到所有致病的FBN1变异。其中包括错义单核苷酸变异（SNV）、全外显子缺失和预测的停止增益SNV。发现FBN1基因和等位基因表达异常。总的来说，MFS中失调的基因在甘油磷脂代谢、免疫、钾通道和细胞外基质过程中富集。单细胞聚类分析确定了SMCs亚型。在MFS中，一些基因在多个SMC亚型（如TRPV2）中有差异表达，而另一些基因在特定SMC状态下有显著性表达（如表达炎症标志物的SMC中的TGFB2）。结论smrna -seq分析能准确鉴定出FBN1变异。确定了对等位基因和基因表达的一般和患者特异性影响。MFS患者主动脉SMCs中甘油磷脂代谢可能失调。通过转录组分析鉴定病原特征可以指导新的诊断和治疗策略。

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引用次数: 0

Bioinformatics tools for drug repurposing: a tutorial using heart failure as a case study 药物再利用的生物信息学工具：以心力衰竭为案例研究的教程

Journal of molecular and cellular cardiology plus

Pub Date : 2025-06-16 DOI: 10.1016/j.jmccpl.2025.100460

Ivo Fonseca , Fábio Trindade , Mário Santos , Adelino Leite-Moreira , Daniel Moreira-Gonçalves , Rui Vitorino , Rita Ferreira , Rita Nogueira-Ferreira

Purpose

Drug repurposing is a crucial strategy for researchers worldwide to accelerate drug development and mitigate associated risks and costs. Heart failure (HF) is a major global health problem with high prevalence and mortality rates. There are significant sex differences in HF, including in the risk factors and phenotype, which demand a sex-personalized drug treatment. A convenient approach in that direction is the reuse of drugs already approved for other conditions that are known to interact in sex-biased dysregulated pathways in HF. Numerous bioinformatics tools can help identify those candidates. This tutorial explores the utility of specific bioinformatics tools in identifying drugs to treat HF as a case study.

Methods

Herein, we explain how NeDRex helps identify genes associated with disease and drug repurposing and how functional enrichment analysis can be performed with ShinyGO. We also explain how to predict targets of small bioactive molecules with SwissTargetPrediction and how to retrieve known and predicted interactions between chemicals and proteins with STITCH.

Results

The tutorial demonstrates the use of these tools in searching for new HF treatments.

Conclusion

This tutorial is designed to ease entry into the utilization of the mentioned bioinformatics tools. This approach can also set a precedent for applying such tools to other diseases. The results presented in this tutorial are illustrative and do not constitute definitive evidence. They are intended for demonstration purposes only.

药物再利用是全球研究人员加速药物开发和降低相关风险和成本的关键策略。心力衰竭是一个主要的全球健康问题，发病率和死亡率都很高。心衰存在显著的性别差异，包括危险因素和表型，这需要性别个性化的药物治疗。在这个方向上，一个方便的方法是重复使用已经被批准用于其他疾病的药物，这些药物已知会在心衰的性别偏倚失调途径中相互作用。许多生物信息学工具可以帮助识别这些候选者。本教程以案例研究为例，探讨了特定生物信息学工具在识别治疗心衰药物方面的应用。方法在本文中，我们解释了NeDRex如何帮助识别与疾病和药物再利用相关的基因，以及如何使用ShinyGO进行功能富集分析。我们还解释了如何使用SwissTargetPrediction预测小生物活性分子的靶标，以及如何使用STITCH检索已知和预测的化学物质与蛋白质之间的相互作用。结果本教程演示了这些工具在寻找新的心衰治疗方法中的应用。本教程的目的是为了方便入门使用上述生物信息学工具。这种方法还可以为将此类工具应用于其他疾病开创先例。本教程中提供的结果是说明性的，并不构成确定的证据。它们仅用于演示目的。

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引用次数: 0

Characterization of human living myocardial slices culture-induced adaptations: a translational perspective 人类活体心肌切片培养诱导适应的表征：翻译视角

Journal of molecular and cellular cardiology plus

Pub Date : 2025-06-12 DOI: 10.1016/j.jmccpl.2025.100465

Jort S.A. van der Geest , Ernest Diez Benavente , Willem B. van Ham , Pieter A. Doevendans , Linda W. van Laake , Teun P. de Boer , Vasco Sampaio-Pinto , Joost P.G. Sluijter

Heart failure involves complex pathophysiological processes, best studied in multicellular human cardiac tissues that reflect the native cellular composition and microenvironment. However, maintaining primary cells and tissues in culture for extended periods remains challenging. Developing robust human cardiac models is critical for advancing preclinical research and bridging the gap to clinical applications. This study aims to characterize adaptations occurring in human living myocardial slices (LMS) during ex vivo culture.

During culture, LMS demonstrated progressive enhancements in contractile function including stronger force generation, reduced diastolic tension, and faster contraction-relaxation kinetics. However, excitability and force-frequency response decreased over the same period. Cultured LMS showed enhanced calcium handling, including increased ability to follow pacing, higher amplitude, and faster, more stable calcium re-uptake. Structurally, LMS displayed no changes in sarcomeres, cell-cell connections, or mitochondria, despite gene expression changes in cytoskeletal and extracellular matrix-related pathways. Transcriptomic analysis revealed metabolic activation with upregulation of metabolism-related pathways. Interestingly, LMS exhibited increased expression of genes associated with early cardiac development after the culture period.

LMS provide a powerful translational model for cardiovascular research, enabling the evaluation of novel therapies and fundamental studies. However, culture-induced adaptations must be carefully considered when interpreting results to ensure physiological and disease relevance.

心力衰竭涉及复杂的病理生理过程，在反映天然细胞组成和微环境的多细胞人类心脏组织中得到了最好的研究。然而，在长时间的培养中维持原代细胞和组织仍然具有挑战性。开发健壮的人类心脏模型对于推进临床前研究和弥合临床应用的差距至关重要。本研究旨在描述人类活体心肌切片（LMS）在离体培养过程中发生的适应性。在培养过程中，LMS表现出收缩功能的渐进式增强，包括更强的力产生、舒张张力降低和更快的收缩-松弛动力学。然而，兴奋性和力频响应在同一时期下降。培养的LMS显示钙处理能力增强，包括跟随起搏的能力增强，振幅更高，钙再摄取更快，更稳定。在结构上，尽管细胞骨架和细胞外基质相关通路的基因表达发生了变化，但LMS在肌瘤、细胞-细胞连接或线粒体方面没有变化。转录组学分析显示代谢激活与代谢相关通路上调。有趣的是，LMS在培养期后表现出与早期心脏发育相关的基因表达增加。LMS为心血管研究提供了一个强大的转化模型，使新疗法和基础研究的评估成为可能。然而，在解释结果时必须仔细考虑文化诱导的适应，以确保生理和疾病的相关性。

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引用次数: 0

Pericellular oxygen dynamics in human cardiac fibroblasts and iPSC-cardiomyocytes in high-throughput plates: insights from experiments and modeling 高通量板中人类心脏成纤维细胞和ipsc -心肌细胞的细胞周氧动力学：来自实验和建模的见解

Journal of molecular and cellular cardiology plus

Pub Date : 2025-06-11 DOI: 10.1016/j.jmccpl.2025.100464

Weizhen Li , David McLeod , Sarah Antonevich, Maria R. Pozo, Zhenyu Li, Emilia Entcheva

Adequate oxygen supply is crucial for proper cellular function. The emergence of high-throughput (HT) expansion of human stem-cell-derived cells and HT in vitro cellular assays for drug testing necessitate monitoring and understanding of the oxygenation conditions, yet virtually no data exists for such settings. We used HT label-free optical measurements and computational modeling to gain insights about oxygen availability (pericellular oxygen dynamics) in syncytia of human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CM) and human cardiac fibroblasts (cFB) grown in glass-bottom 96-well plates under static conditions. Our experimental results highlight the critical role of cell density and solution height (oxygen delivery path) in pericellular oxygen dynamics. The developed computational model, trained on the obtained comprehensive data set, revealed that time-variant maximum oxygen consumption rate, V_max, is needed to faithfully capture the complex pericellular oxygen dynamics in the excitable hiPSC-CMs, but not in the cFBs. Interestingly, hypoxia (<2 % pericellular oxygen) developed within hours in the dense iPSC-CM cultures when the solution volume was sufficiently large. Conversely, hiPSC-CMs grown at low cell density or in smaller solution volume, as well as cFB under all studied conditions, were found to operate in hyperoxic (>7 %) conditions. Pericellular oxygen dynamics of the differentiated hiPSC-CMs evolved over days in culture, with the best improvement in respiration seen in samples operating close to normoxia. Our results and the developed computational model can be used directly to optimize cardiac cell growth in HT plates and achieve desired physiological conditions, which is important in cellular assays for cardiotoxicity, drug development, personalized medicine and heart regeneration applications.

充足的氧气供应对正常的细胞功能至关重要。人类干细胞衍生细胞的高通量（HT）扩增和用于药物测试的体外HT细胞分析的出现需要监测和了解氧合条件，但实际上没有这样的数据存在。我们使用无HT标记的光学测量和计算模型来了解在静态条件下生长的人诱导多能干细胞来源的心肌细胞（hiPSC-CM）和人心脏成纤维细胞（cFB）合胞体中的氧可用性（细胞外氧动力学）。我们的实验结果强调了细胞密度和溶液高度（氧气输送路径）在细胞周氧动力学中的关键作用。建立的计算模型在获得的综合数据集上进行了训练，结果表明，在可激发的hiPSC-CMs中，需要时变最大耗氧量（Vmax）来忠实地捕捉复杂的细胞外氧动力学，而在cfb中则不需要。有趣的是，当溶液体积足够大时，在密集的iPSC-CM培养中，缺氧（2%细胞外氧）在数小时内发生。相反，在低细胞密度或较小溶液体积下生长的hiPSC-CMs，以及在所有研究条件下的cFB，都被发现在高氧（> 7%）条件下运行。分化的hiPSC-CMs的细胞外氧动力学在培养过程中经过数天的进化，在接近常氧环境的样品中，呼吸得到了最好的改善。我们的结果和开发的计算模型可以直接用于优化HT板中的心脏细胞生长并达到所需的生理条件，这在心脏毒性，药物开发，个性化医疗和心脏再生应用的细胞分析中具有重要意义。

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