Acute lung injury (ALI), especially when resulting from trauma-associated hemorrhagic shock (THS), is a life-threatening condition with limited treatment options and high mortality. Understanding the molecular mechanisms driving ALI in this context is essential to identify reliable biomarkers and therapeutic targets. This study aimed to explore the transcriptomic alterations and protein interaction networks in a rat model of THS-induced ALI using RNA sequencing and bioinformatics tools. RNA-seq analysis was performed on lung tissues from five THS-induced and five normal rats. Analysis revealed 1003 differentially expressed genes, including 365 upregulated and 638 downregulated. Functional enrichment pointed to significant involvement of pathways related to oxidative stress, hypoxia response, neutrophil degranulation, ferroptosis, and immune activation. Protein-protein interaction network analysis identified four key gene modules, with Module 3 notably associated with iron metabolism and neutrophilic inflammation. Hub genes such as Cd163, Nqo1, Gclc, Lcn2, and Mmp8 were identified as central regulators and validated in independent samples (three THS-induced and three controls). Lcn2 and cathepsins (CTSS, CTSK, CTSL) emerged as particularly relevant for their multifaceted roles in inflammation, iron homeostasis, and matrix remodeling. These findings provide novel insights into the immunometabolic dysregulation underlying THS-induced ALI and suggest promising molecular targets for future therapeutic interventions aimed at mitigating lung injury in critically injured trauma patients.
{"title":"Unveiling the molecular mechanisms of hemorrhagic shock and acute lung injury: An integrative RNA-Seq and network analysis","authors":"Manjaree Mishra , Shivangi Agrawal , Shashi P. Mishra , Rajiv Kumar , Katyayani Mishra , Ekta Pathak , Rajeev Mishra","doi":"10.1016/j.trsl.2025.10.002","DOIUrl":"10.1016/j.trsl.2025.10.002","url":null,"abstract":"<div><div>Acute lung injury (ALI), especially when resulting from trauma-associated hemorrhagic shock (THS), is a life-threatening condition with limited treatment options and high mortality. Understanding the molecular mechanisms driving ALI in this context is essential to identify reliable biomarkers and therapeutic targets. This study aimed to explore the transcriptomic alterations and protein interaction networks in a rat model of THS-induced ALI using RNA sequencing and bioinformatics tools. RNA-seq analysis was performed on lung tissues from five THS-induced and five normal rats. Analysis revealed 1003 differentially expressed genes, including 365 upregulated and 638 downregulated. Functional enrichment pointed to significant involvement of pathways related to oxidative stress, hypoxia response, neutrophil degranulation, ferroptosis, and immune activation. Protein-protein interaction network analysis identified four key gene modules, with Module 3 notably associated with iron metabolism and neutrophilic inflammation. Hub genes such as Cd163, Nqo1, Gclc, Lcn2, and Mmp8 were identified as central regulators and validated in independent samples (three THS-induced and three controls). Lcn2 and cathepsins (CTSS, CTSK, CTSL) emerged as particularly relevant for their multifaceted roles in inflammation, iron homeostasis, and matrix remodeling. These findings provide novel insights into the immunometabolic dysregulation underlying THS-induced ALI and suggest promising molecular targets for future therapeutic interventions aimed at mitigating lung injury in critically injured trauma patients.</div></div>","PeriodicalId":23226,"journal":{"name":"Translational Research","volume":"284 ","pages":"Pages 17-28"},"PeriodicalIF":5.9,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145411395","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-01Epub Date: 2025-08-27DOI: 10.1016/j.trsl.2025.08.004
Elisa Gambini , Erica Rurali , Veronica Barbagallo , Sergio Pirola , Alessandro Scopece , Andrea Biondi , Beatrice Bassetti , Manuel Casaburo , Luana Eramo , Giorgio Pio Alberto Marinelli , Diego Farinello , Simona Rodighiero , Yuri D’alessandra , Mattia Chiesa , Gabriella Spaltro , Veronica Ricci , Aoife Gowran , Elisa Castiglioni , Daniele Fileccia , Giuseppe Nanci , Giulio Pompilio
Background
Despite recent significant therapeutic progress, cardiovascular diseases (CVD) remain an unmet clinical, economic, and social burden worldwide. Cell-based therapies have been proposed as therapeutic strategies, however, the overall efficacy was modest.
Objective
We aimed to fully characterize a novel subpopulation of CD90− mesenchymal cells derived from human heart tissue (hCmPC90-) and evaluate its ability to induce cardiac tissue repair and functional recovery.
Methods
We performed a comprehensive phenotypic characterization of the hCmPC90− by flow cytometry and RNA sequencing. A direct comparison of hCmPC90− with previously clinically tested bone marrow- and cardiac-derived cell types, has been conducted both in vitro by means of various assays of angiogenic potency, and in vivo, by testing the ability to ameliorate left ventricular function in a mouse model of acute myocardial infarction (AMI).
Results
hCmPC90− showed distinct surface markers and transcriptional phenotype compared with unselected mesenchymal heart cells (hCmPCs) and the positive CD90 counterpart (hCmPC90+). When human hCmPC90−, hCmPC90+, hCmPC, cardiosphere-derived cells (CDCs), and bone marrow-derived CD34+ cells were functionally tested in vitro, hCmPC90− revealed a superior endothelial differentiation ability, higher anti-inflammatory, cardio-protective capacity, and angiocrine activity. Moreover, hCmPC90− showed specific immune-privileged features. When intramyocardially delivered into infarcted mouse hearts, hCmPC90− outperformed three weeks after injection other clinical-grade cell types, as for left ventricular (LV) function and adverse LV remodeling recovery, infarct size reduction, and vascular density augmentation.
Conclusion
hCmPC90− shows a superior biological potency which deserves clinical exploitation as an advanced therapy medicinal product in the context of refractory ischemic heart disease.
{"title":"Angiogenic and reparative potency of a human cardiac CD90− mesenchymal subpopulation in heart ischemic model","authors":"Elisa Gambini , Erica Rurali , Veronica Barbagallo , Sergio Pirola , Alessandro Scopece , Andrea Biondi , Beatrice Bassetti , Manuel Casaburo , Luana Eramo , Giorgio Pio Alberto Marinelli , Diego Farinello , Simona Rodighiero , Yuri D’alessandra , Mattia Chiesa , Gabriella Spaltro , Veronica Ricci , Aoife Gowran , Elisa Castiglioni , Daniele Fileccia , Giuseppe Nanci , Giulio Pompilio","doi":"10.1016/j.trsl.2025.08.004","DOIUrl":"10.1016/j.trsl.2025.08.004","url":null,"abstract":"<div><h3>Background</h3><div>Despite recent significant therapeutic progress, cardiovascular diseases (CVD) remain an unmet clinical, economic, and social burden worldwide. Cell-based therapies have been proposed as therapeutic strategies, however, the overall efficacy was modest.</div></div><div><h3>Objective</h3><div>We aimed to fully characterize a novel subpopulation of CD90<sup>−</sup> mesenchymal cells derived from human heart tissue (hCmPC90<sup>-</sup>) and evaluate its ability to induce cardiac tissue repair and functional recovery.</div></div><div><h3>Methods</h3><div>We performed a comprehensive phenotypic characterization of the hCmPC90<sup>−</sup> by flow cytometry and RNA sequencing. A direct comparison of hCmPC90<sup>−</sup> with previously clinically tested bone marrow- and cardiac-derived cell types, has been conducted both <em>in vitro</em> by means of various assays of angiogenic potency, and <em>in vivo,</em> by testing the ability to ameliorate left ventricular function in a mouse model of acute myocardial infarction (AMI).</div></div><div><h3>Results</h3><div>hCmPC90<sup>−</sup> showed distinct surface markers and transcriptional phenotype compared with unselected mesenchymal heart cells (hCmPCs) and the positive CD90 counterpart (hCmPC90<sup>+</sup>). When human hCmPC90<sup>−</sup>, hCmPC90<sup>+</sup>, hCmPC, cardiosphere-derived cells (CDCs), and bone marrow-derived CD34<sup>+</sup> cells were functionally tested <em>in vitro</em>, hCmPC90<sup>−</sup> revealed a superior endothelial differentiation ability, higher anti-inflammatory, cardio-protective capacity, and angiocrine activity. Moreover, hCmPC90<sup>−</sup> showed specific immune-privileged features. When intramyocardially delivered into infarcted mouse hearts, hCmPC90<sup>−</sup> outperformed three weeks after injection other clinical-grade cell types, as for left ventricular (LV) function and adverse LV remodeling recovery, infarct size reduction, and vascular density augmentation.</div></div><div><h3>Conclusion</h3><div>hCmPC90<sup>−</sup> shows a superior biological potency which deserves clinical exploitation as an advanced therapy medicinal product in the context of refractory ischemic heart disease.</div></div>","PeriodicalId":23226,"journal":{"name":"Translational Research","volume":"283 ","pages":"Pages 22-35"},"PeriodicalIF":5.9,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144984658","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-01Epub Date: 2025-09-11DOI: 10.1016/j.trsl.2025.09.001
Yu Zhang , Shan Cao , Yun Liu , Ziliang Ke , Zhe Wu , Xiaohui Fang , Yang Zhang , Jingyi Chen , Congyi Yang , Yiken Lin , Ning Chen , Jun Xu , Yulan Liu
The appendix, a component of the gut-associated lymphoid tissue enriched with B lymphocytes, plays a pivotal role in intestinal mucosal immunity. Previous studies have indicated that prior appendectomy may prevent the onset of ulcerative colitis (UC); however, its therapeutic role in UC remains unclear, and prophylactic appendectomy is not a realistic approach to prevent UC. In this study, we confirmed that appendectomy alleviates dextran sodium sulphate (DSS)-induced chronic murine colitis and further demonstrated that appendiceal B (APB) lymphocytes exacerbate colonic inflammation by migrating to the colon via the CCL20–CCR6 axis and facilitating colonic CD4+ T cell-mediated T helper 1 (Th1) and T helper 17 (Th17) immune responses. Single-cell sequencing of colonic tissues revealed IgG+ B cell-skewed responses in patients with UC, and APB cell expansion was positively correlated with disease severity. Immunofluorescence co-staining suggested that colonic B cells of UC patients related to the appendix. These findings highlight the therapeutic potential of appendectomy and B cell-targeted immunotherapy in UC treatment and further introduce the hypothesis that UC with appendiceal orifice inflammation may represent a distinct subtype of the disease.
{"title":"Appendiceal B lymphocytes contribute to the pathogenesis of experimental colitis through fueling colonic CD4+ T polarization","authors":"Yu Zhang , Shan Cao , Yun Liu , Ziliang Ke , Zhe Wu , Xiaohui Fang , Yang Zhang , Jingyi Chen , Congyi Yang , Yiken Lin , Ning Chen , Jun Xu , Yulan Liu","doi":"10.1016/j.trsl.2025.09.001","DOIUrl":"10.1016/j.trsl.2025.09.001","url":null,"abstract":"<div><div>The appendix, a component of the gut-associated lymphoid tissue enriched with B lymphocytes, plays a pivotal role in intestinal mucosal immunity. Previous studies have indicated that prior appendectomy may prevent the onset of ulcerative colitis (UC); however, its therapeutic role in UC remains unclear, and prophylactic appendectomy is not a realistic approach to prevent UC. In this study, we confirmed that appendectomy alleviates dextran sodium sulphate (DSS)-induced chronic murine colitis and further demonstrated that appendiceal B (APB) lymphocytes exacerbate colonic inflammation by migrating to the colon via the CCL20–CCR6 axis and facilitating colonic CD4<sup>+</sup> T cell-mediated T helper 1 (Th1) and T helper 17 (Th17) immune responses. Single-cell sequencing of colonic tissues revealed IgG<sup>+</sup> B cell-skewed responses in patients with UC, and APB cell expansion was positively correlated with disease severity. Immunofluorescence co-staining suggested that colonic B cells of UC patients related to the appendix. These findings highlight the therapeutic potential of appendectomy and B cell-targeted immunotherapy in UC treatment and further introduce the hypothesis that UC with appendiceal orifice inflammation may represent a distinct subtype of the disease.</div></div>","PeriodicalId":23226,"journal":{"name":"Translational Research","volume":"283 ","pages":"Pages 56-70"},"PeriodicalIF":5.9,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145058859","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-01Epub Date: 2025-09-09DOI: 10.1016/j.trsl.2025.09.002
Ming-Mei Shang , Zhuang Liu , Bogdan Knezevic , Christine Möller Westerberg , Sudeepta Kumar Panda , Hai Fang , Ning Xu Landén , Michael Sundström , Julian C. Knight , Louise Berg
Background and Purpose
Systemic lupus erythematosus (SLE) is a systemic autoimmune disease involving production of autoantibodies by B cells. This study aimed at identifying novel drug targets using a computational algorithm to select targets and thereafter validate the top ranked 11 targets by siRNA knockdown in a primary B cell maturation assay.
Experimental Approach
The top 1 % genes (∼150 genes) from SLE genome-wide association studies were ranked by Priority index (Pi), a computational tool integrating genomic and network information to prioritize disease-relevant genes. These were further filtered by network connectivity, drugability, for ranking highly in autoimmune diseases and for not directly interfering with the B cell stimulation cocktail used. From this, 11 genes were selected for validation by siRNA knockdown: IFNGR1, IL-2, IRF4, IL-12A, IL-12B, VCAM-1, ATF6B, RELA, IKBKG, CHUK and MAPK14. Effects on induced maturation and viability of primary blood B cells were analyzed by flow cytometry, and effects on IgG secretion were investigated by ELISA. RNA-sequencing of B cells treated with siRNA was performed to investigate molecular mechanisms underlying the functional alterations.
Key Results
Experimental results show that several of the targets (IFNGR1, IL-2, IL-12A, MAPK14, IRF4, CHUK, ATF6B, IKBKG, and RELA) are involved in B cell maturation, as knockdown caused reduced IgG production and/or decreased maturation of B cells. The observed variability of effects on IgG secretion and B cell maturation suggests differences in the mechanistic roles of the proteins encoded by these genes. RNA-seq analysis of cells where expression of the targeted genes had been modulated showed effects on the expression level of hundreds of genes involved in cellular processes important for B cell functions.
Conclusion and Implications
Combining the target prioritization algorithm with experimental functional validation studies by gene knockdown and whole transcriptomics profiling constitutes a promising approach to identify potential novel drug targets in immune disorders.
{"title":"Combining computational target prioritization and a B cell maturation assay for target evaluation studies in systemic lupus erythematosus","authors":"Ming-Mei Shang , Zhuang Liu , Bogdan Knezevic , Christine Möller Westerberg , Sudeepta Kumar Panda , Hai Fang , Ning Xu Landén , Michael Sundström , Julian C. Knight , Louise Berg","doi":"10.1016/j.trsl.2025.09.002","DOIUrl":"10.1016/j.trsl.2025.09.002","url":null,"abstract":"<div><h3>Background and Purpose</h3><div>Systemic lupus erythematosus (SLE) is a systemic autoimmune disease involving production of autoantibodies by B cells. This study aimed at identifying novel drug targets using a computational algorithm to select targets and thereafter validate the top ranked 11 targets by siRNA knockdown in a primary B cell maturation assay.</div></div><div><h3>Experimental Approach</h3><div>The top 1 % genes (∼150 genes) from SLE genome-wide association studies were ranked by Priority index (Pi), a computational tool integrating genomic and network information to prioritize disease-relevant genes. These were further filtered by network connectivity, drugability, for ranking highly in autoimmune diseases and for not directly interfering with the B cell stimulation cocktail used. From this, 11 genes were selected for validation by siRNA knockdown: <em>IFNGR1, IL-2, IRF4, IL-12A, IL-12B, VCAM-1, ATF6B, RELA, IKBKG, CHUK</em> and <em>MAPK14.</em> Effects on induced maturation and viability of primary blood B cells were analyzed by flow cytometry, and effects on IgG secretion were investigated by ELISA. RNA-sequencing of B cells treated with siRNA was performed to investigate molecular mechanisms underlying the functional alterations.</div></div><div><h3>Key Results</h3><div>Experimental results show that several of the targets (<em>IFNGR1, IL-2, IL-12A, MAPK14, IRF4, CHUK, ATF6B, IKBKG,</em> and <em>RELA</em>) are involved in B cell maturation, as knockdown caused reduced IgG production and/or decreased maturation of B cells. The observed variability of effects on IgG secretion and B cell maturation suggests differences in the mechanistic roles of the proteins encoded by these genes. RNA-seq analysis of cells where expression of the targeted genes had been modulated showed effects on the expression level of hundreds of genes involved in cellular processes important for B cell functions.</div></div><div><h3>Conclusion and Implications</h3><div>Combining the target prioritization algorithm with experimental functional validation studies by gene knockdown and whole transcriptomics profiling constitutes a promising approach to identify potential novel drug targets in immune disorders.</div></div>","PeriodicalId":23226,"journal":{"name":"Translational Research","volume":"283 ","pages":"Pages 36-46"},"PeriodicalIF":5.9,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145042674","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-01Epub Date: 2025-09-19DOI: 10.1016/j.trsl.2025.09.003
Min-Jung Park , Junhyeong Lee , Merc Emil Matienzo , Sangyi Lim , Keon Kim , Chang-Min Lee , Dong-il Kim
Adaptive thermogenesis, particularly via the β3-adrenergic receptor (ADRB3)–protein kinase A catalytic subunit α (PKA Cα)–uncoupling protein 1 (UCP1) pathway, promotes energy expenditure and contributes to metabolic homeostasis, thereby establishing this pathway as a promising therapeutic target for metabolic disorders associated with excessive energy intake. In this study, we aimed to evaluate the therapeutic potential of adeno-associated virus (AAV)-mediated gene therapy targeting thermogenic pathways in metabolic tissues for the treatment of obesity-related dysfunctions. We demonstrated that adipocyte-specific overexpression of UCP1 improved glucose tolerance. Similarly, ADRB3 overexpression significantly enhanced glucose tolerance. Furthermore, ectopic expression of UCP1 in hepatocytes and myofibers also led to improved glucose tolerance. These findings highlight the potential of AAV-mediated gene therapy targeting the ADRB3–PKA Cα–UCP1 axis as a promising strategy for the treatment of obesity-associated metabolic disorders.
{"title":"Delivery of thermogenic genes to metabolic tissues: Effects on body weight and glucose tolerance","authors":"Min-Jung Park , Junhyeong Lee , Merc Emil Matienzo , Sangyi Lim , Keon Kim , Chang-Min Lee , Dong-il Kim","doi":"10.1016/j.trsl.2025.09.003","DOIUrl":"10.1016/j.trsl.2025.09.003","url":null,"abstract":"<div><div>Adaptive thermogenesis, particularly via the β3-adrenergic receptor (ADRB3)–protein kinase A catalytic subunit α (PKA Cα)–uncoupling protein 1 (UCP1) pathway, promotes energy expenditure and contributes to metabolic homeostasis, thereby establishing this pathway as a promising therapeutic target for metabolic disorders associated with excessive energy intake. In this study, we aimed to evaluate the therapeutic potential of adeno-associated virus (AAV)-mediated gene therapy targeting thermogenic pathways in metabolic tissues for the treatment of obesity-related dysfunctions. We demonstrated that adipocyte-specific overexpression of UCP1 improved glucose tolerance. Similarly, ADRB3 overexpression significantly enhanced glucose tolerance. Furthermore, ectopic expression of UCP1 in hepatocytes and myofibers also led to improved glucose tolerance. These findings highlight the potential of AAV-mediated gene therapy targeting the ADRB3–PKA Cα–UCP1 axis as a promising strategy for the treatment of obesity-associated metabolic disorders.</div></div>","PeriodicalId":23226,"journal":{"name":"Translational Research","volume":"283 ","pages":"Pages 47-55"},"PeriodicalIF":5.9,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145115803","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号