Current therapies for inflammatory bowel disease(IBD)often fail to achieve complete remission and are associated with systemic toxicity owing to their broad immunosuppressive effects.To overcome these limitations,we d...Current therapies for inflammatory bowel disease(IBD)often fail to achieve complete remission and are associated with systemic toxicity owing to their broad immunosuppressive effects.To overcome these limitations,we developed a bioengineered extracellular vesicle(EV)platform that modulates key immune signaling pathways to efficiently restore the T-cell balance in inflamed intestinal tissues.EVs derived from Wharton’s jelly mesenchymal stem cells were engineered to display PD-L1 on their surface and encapsulate miR-27a-3p.Surface PD-L1 engages the PD-1 checkpoint in activated T cells,attenuating T-cell receptor signaling via SHP2-mediated dephosphorylation of ZAP70 and AKT.In parallel,miR-27a-3p suppresses prohibitin 1(PHB1),a mitochondrial regulator of Th17 cell bioenergetics and inflammatory function,thereby reducing Th17 polarization and increasing the number of FOXP3⁺regulatory T cells.These dual-targeting EVs preferentially localized to inflamed intestinal tissues via chemokine(CCR2/CXCR4)and PD-1-dependent mechanisms.In humanized mouse models of colitis,these EVs attenuated mucosal inflammation,suppressed effector T-cell responses,and preserved epithelial integrity.In IBD patient-derived colonoid cultures,PDL1/miR-27a-3p EVs maintained epithelial viability and barrier integrity without inducing cytotoxicity or structural disruption.Transcriptomic and single-cell analyses revealed the downregulation of inflammatory and exhaustion signatures,along with the enrichment of regulatory subsets.Collectively,this study presents a cell-free immunotherapeutic approach that reprograms T cells in inflamed tissues through the PD-1 and mitochondrial signaling pathways while maintaining intestinal epithelial integrity,offering a promising therapeutic strategy for IBD and other T cell-driven inflammatory disorders.展开更多
Background Tetralogy of Fallot （TOF） is the most common malformation of children with an incidence of approximately 10% of congenital heart disease patients. There can be a wide spectrum to the severity of the anato...Background Tetralogy of Fallot （TOF） is the most common malformation of children with an incidence of approximately 10% of congenital heart disease patients. There can be a wide spectrum to the severity of the anatomic defects, which include ventricular septal defect, aortic override, right ventricular outflow tract obstruction, and right ventricular hypertrophy. We examined the relationship between right ventricular hypertrophy in patients with TOF and the gene expression of factors in the mitogen-activated protein kinase （MAPK） signal pathway. Methods To gain insight into the characteristic gene（s） involved in molecular mechanisms of right ventricular hypertrophy in TOF, differential mRNA and micro RNA expression profiles were assessed using expression-based micro array technology on right ventricular biopsies from young TOF patients who underwent primary correction and on normal heart tissue. We then analyzed the gene expression of the MAPK signal pathway using reverse transcription-polymerase chain reaction （RT-PCR） in normals and TOF patients. Results Using the micro RNA chip V3.0 and human whole genome oligonucleotide microarray VI.0 to detect the gene expression, we found 1068 genes showing altered expression of at least two-fold in TOF patients compared to the normal hearts, and 47 micro RNAs that showed a significant difference of at least two-fold in TOF patients. We then analyzed these mRNAs and micro RNAs by target gene predicting software Microcosm Targets version 5.0, and determined those mRNA highly relevant to the right ventricular hypertrophy by RT-PCR method. There were obvious differences in the gene expression of factors in the MAPK signal pathway when using RT-PCR, which was consistent to the results of the cDNA microarray.Conclusion The upregulation of genes in the MAPK signal pathway may be the key events that contribute to right ventricular hypertrophy and stunted angiogenesis in patients with TOF.展开更多
基金supported by the Basic Research Laboratory Program(Advanced Type)through the National Research Foundation of Korea(NRF),funded by the Korean Government(RS-2023-00218476)Image created with BioRender.com with permission.
文摘Current therapies for inflammatory bowel disease(IBD)often fail to achieve complete remission and are associated with systemic toxicity owing to their broad immunosuppressive effects.To overcome these limitations,we developed a bioengineered extracellular vesicle(EV)platform that modulates key immune signaling pathways to efficiently restore the T-cell balance in inflamed intestinal tissues.EVs derived from Wharton’s jelly mesenchymal stem cells were engineered to display PD-L1 on their surface and encapsulate miR-27a-3p.Surface PD-L1 engages the PD-1 checkpoint in activated T cells,attenuating T-cell receptor signaling via SHP2-mediated dephosphorylation of ZAP70 and AKT.In parallel,miR-27a-3p suppresses prohibitin 1(PHB1),a mitochondrial regulator of Th17 cell bioenergetics and inflammatory function,thereby reducing Th17 polarization and increasing the number of FOXP3⁺regulatory T cells.These dual-targeting EVs preferentially localized to inflamed intestinal tissues via chemokine(CCR2/CXCR4)and PD-1-dependent mechanisms.In humanized mouse models of colitis,these EVs attenuated mucosal inflammation,suppressed effector T-cell responses,and preserved epithelial integrity.In IBD patient-derived colonoid cultures,PDL1/miR-27a-3p EVs maintained epithelial viability and barrier integrity without inducing cytotoxicity or structural disruption.Transcriptomic and single-cell analyses revealed the downregulation of inflammatory and exhaustion signatures,along with the enrichment of regulatory subsets.Collectively,this study presents a cell-free immunotherapeutic approach that reprograms T cells in inflamed tissues through the PD-1 and mitochondrial signaling pathways while maintaining intestinal epithelial integrity,offering a promising therapeutic strategy for IBD and other T cell-driven inflammatory disorders.
文摘Background Tetralogy of Fallot （TOF） is the most common malformation of children with an incidence of approximately 10% of congenital heart disease patients. There can be a wide spectrum to the severity of the anatomic defects, which include ventricular septal defect, aortic override, right ventricular outflow tract obstruction, and right ventricular hypertrophy. We examined the relationship between right ventricular hypertrophy in patients with TOF and the gene expression of factors in the mitogen-activated protein kinase （MAPK） signal pathway. Methods To gain insight into the characteristic gene（s） involved in molecular mechanisms of right ventricular hypertrophy in TOF, differential mRNA and micro RNA expression profiles were assessed using expression-based micro array technology on right ventricular biopsies from young TOF patients who underwent primary correction and on normal heart tissue. We then analyzed the gene expression of the MAPK signal pathway using reverse transcription-polymerase chain reaction （RT-PCR） in normals and TOF patients. Results Using the micro RNA chip V3.0 and human whole genome oligonucleotide microarray VI.0 to detect the gene expression, we found 1068 genes showing altered expression of at least two-fold in TOF patients compared to the normal hearts, and 47 micro RNAs that showed a significant difference of at least two-fold in TOF patients. We then analyzed these mRNAs and micro RNAs by target gene predicting software Microcosm Targets version 5.0, and determined those mRNA highly relevant to the right ventricular hypertrophy by RT-PCR method. There were obvious differences in the gene expression of factors in the MAPK signal pathway when using RT-PCR, which was consistent to the results of the cDNA microarray.Conclusion The upregulation of genes in the MAPK signal pathway may be the key events that contribute to right ventricular hypertrophy and stunted angiogenesis in patients with TOF.
