Single-cell atlas reveals multi-faced responses of losartan on tubular mitochondria in diabetic kidney disease
Abstract Background and Objective Mitochondria are crucial to the function of renal tubular cells, and their dynamic perturbation in many aspects is an important mechanism of diabetic kidney disease (DKD). Single-nucleus RNA sequencing (snRNA-seq) technology is a high-throughput sequencing analysis...
Saved in:
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-01-01
|
| Series: | Journal of Translational Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12967-025-06074-5 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832585409525186560 |
|---|---|
| author | Zhen Zhu Guangxin Luan Song Wu Yiyi Song Shuang Shen Kaiyue Wu Shengnan Qian Weiping Jia Jun Yin Tao Ren Jianping Ye Li Wei |
| author_facet | Zhen Zhu Guangxin Luan Song Wu Yiyi Song Shuang Shen Kaiyue Wu Shengnan Qian Weiping Jia Jun Yin Tao Ren Jianping Ye Li Wei |
| author_sort | Zhen Zhu |
| collection | DOAJ |
| description | Abstract Background and Objective Mitochondria are crucial to the function of renal tubular cells, and their dynamic perturbation in many aspects is an important mechanism of diabetic kidney disease (DKD). Single-nucleus RNA sequencing (snRNA-seq) technology is a high-throughput sequencing analysis technique for RNA at the level of a single cell nucleus. Here, our DKD mouse kidney single-cell RNA sequencing conveys a more comprehensive mitochondrial profile, which helps us further understand the therapeutic response of this unique organelle family to drugs. Methods After high fat diet (HFD), mice were intraperitoneally injected with streptozotocin (STZ) to induce DKD, and then divided into three subsets: CON (healthy) subset, DKD (vehicle) subset, and LST (losartan; 25 mg/kg/day) subset. Divide HK-2 cell into LG (low glucose; 5 mM) and HG (high glucose; 30 mM) and HG + LST (losartan; 1 µ M) subsets. snRNA-seq was performed on the renal tissues of LST and DKD subset mice. To reveal the effects of losartan on gene function and pathway changes in renal tubular mitochondria, Gene Ontology (GO) enrichment analysis and GSEA/GSVA scoring were performed to analyze the specific response of proximal tubular (PT) cell mitochondria to losartan treatment, including key events in mitochondrial homeostasis such as mitochondrial morphology, dynamics, mitophagy, autophagic flux, mitochondrial respiratory chain, apoptosis, and ROS generation. Preliminary validation through in vitro and in vivo experiments, including observation of changes in mitochondrial morphology and dynamics using probes such as Mitotracker Red, and evaluation of the effect of losartan on key events of mitochondrial homeostasis perturbation using electron microscopy, laser confocal microscopy, immunofluorescence, and Western blotting. Detection of autophagic flux in cells by transfecting Ad-mCherry-GFP-LC3B dual fluorescence labeled adenovirus. Various fluorescent probes and energy detector are used to detect mitochondrial apoptosis, ROS, and respiration of mitochondrion. Results Through the single-cell atlas of DKD mouse kidneys, it was found that losartan treatment significantly increased the percentage of PT cells. Gene Ontology (GO) enrichment analysis of differentially expressed genes showed enrichment of autophagy of mitochondrion pathway. Further GSEA analysis and GSVA scoring revealed that mitophagy and other key mitochondrial perturbation events, such as ROS production, apoptosis, membrane potential, adenosine triphosphate (ATP) synthesis, and mitochondrial dynamics, were involved in the protective mechanism of losartan on PT cells, thereby improving mitochondrial homeostasis. Consistent results were also obtained in mice and cellular experiments. In addition, we highlighted a specific renal tubular subpopulation with mitophagy phenotype found in single-cell data, and preliminarily validated it with co-localization and increased expression of Pink1 and Gclc in kidney specimens of DKD patients treated with losartan. Conclusions Our research suggests that scRNA-seq can reflect the multifaceted mitochondrial landscape of DKD renal tubular cells after drug treatment, and these findings may provide new targets for DKD therapy at the organelle level. |
| format | Article |
| id | doaj-art-28077734b45c48f4ba6854069d4f2a28 |
| institution | Kabale University |
| issn | 1479-5876 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Translational Medicine |
| spelling | doaj-art-28077734b45c48f4ba6854069d4f2a282025-01-26T12:50:26ZengBMCJournal of Translational Medicine1479-58762025-01-0123112610.1186/s12967-025-06074-5Single-cell atlas reveals multi-faced responses of losartan on tubular mitochondria in diabetic kidney diseaseZhen Zhu0Guangxin Luan1Song Wu2Yiyi Song3Shuang Shen4Kaiyue Wu5Shengnan Qian6Weiping Jia7Jun Yin8Tao Ren9Jianping Ye10Li Wei11Department of Respiratory Medicine, Shanghai Sixth People’s Hospital, Affiliated to Shanghai Jiao Tong University School of MedicineDepartment of Clinical Laboratory, Shanghai Sixth People’s Hospital, Affiliated to Shanghai Jiao Tong University School of MedicineDepartment of Cardiothoracic Surgery, Shanghai Sixth People’s Hospital, Affiliated to Shanghai Jiao Tong University School of MedicineDepartment of Respiratory Medicine, Shanghai Sixth People’s Hospital, Affiliated to Shanghai Jiao Tong University School of MedicineShanghai Diabetes Institute, Shanghai Sixth People’s Hospital, Affiliated to Shanghai Jiao Tong University School of MedicineShanghai Diabetes Institute, Shanghai Sixth People’s Hospital, Affiliated to Shanghai Jiao Tong University School of MedicineShanghai Diabetes Institute, Shanghai Sixth People’s Hospital, Affiliated to Shanghai Jiao Tong University School of MedicineShanghai Diabetes Institute, Shanghai Sixth People’s Hospital, Affiliated to Shanghai Jiao Tong University School of MedicineDepartment of Endocrine Medicine, Shanghai Sixth People’s Hospital, Affiliated to Shanghai Jiao Tong University School of MedicineDepartment of Respiratory Medicine, Shanghai Sixth People’s Hospital, Affiliated to Shanghai Jiao Tong University School of MedicineShanghai Diabetes Institute, Shanghai Sixth People’s Hospital, Affiliated to Shanghai Jiao Tong University School of MedicineDepartment of Endocrine Medicine, Shanghai Sixth People’s Hospital, Affiliated to Shanghai Jiao Tong University School of MedicineAbstract Background and Objective Mitochondria are crucial to the function of renal tubular cells, and their dynamic perturbation in many aspects is an important mechanism of diabetic kidney disease (DKD). Single-nucleus RNA sequencing (snRNA-seq) technology is a high-throughput sequencing analysis technique for RNA at the level of a single cell nucleus. Here, our DKD mouse kidney single-cell RNA sequencing conveys a more comprehensive mitochondrial profile, which helps us further understand the therapeutic response of this unique organelle family to drugs. Methods After high fat diet (HFD), mice were intraperitoneally injected with streptozotocin (STZ) to induce DKD, and then divided into three subsets: CON (healthy) subset, DKD (vehicle) subset, and LST (losartan; 25 mg/kg/day) subset. Divide HK-2 cell into LG (low glucose; 5 mM) and HG (high glucose; 30 mM) and HG + LST (losartan; 1 µ M) subsets. snRNA-seq was performed on the renal tissues of LST and DKD subset mice. To reveal the effects of losartan on gene function and pathway changes in renal tubular mitochondria, Gene Ontology (GO) enrichment analysis and GSEA/GSVA scoring were performed to analyze the specific response of proximal tubular (PT) cell mitochondria to losartan treatment, including key events in mitochondrial homeostasis such as mitochondrial morphology, dynamics, mitophagy, autophagic flux, mitochondrial respiratory chain, apoptosis, and ROS generation. Preliminary validation through in vitro and in vivo experiments, including observation of changes in mitochondrial morphology and dynamics using probes such as Mitotracker Red, and evaluation of the effect of losartan on key events of mitochondrial homeostasis perturbation using electron microscopy, laser confocal microscopy, immunofluorescence, and Western blotting. Detection of autophagic flux in cells by transfecting Ad-mCherry-GFP-LC3B dual fluorescence labeled adenovirus. Various fluorescent probes and energy detector are used to detect mitochondrial apoptosis, ROS, and respiration of mitochondrion. Results Through the single-cell atlas of DKD mouse kidneys, it was found that losartan treatment significantly increased the percentage of PT cells. Gene Ontology (GO) enrichment analysis of differentially expressed genes showed enrichment of autophagy of mitochondrion pathway. Further GSEA analysis and GSVA scoring revealed that mitophagy and other key mitochondrial perturbation events, such as ROS production, apoptosis, membrane potential, adenosine triphosphate (ATP) synthesis, and mitochondrial dynamics, were involved in the protective mechanism of losartan on PT cells, thereby improving mitochondrial homeostasis. Consistent results were also obtained in mice and cellular experiments. In addition, we highlighted a specific renal tubular subpopulation with mitophagy phenotype found in single-cell data, and preliminarily validated it with co-localization and increased expression of Pink1 and Gclc in kidney specimens of DKD patients treated with losartan. Conclusions Our research suggests that scRNA-seq can reflect the multifaceted mitochondrial landscape of DKD renal tubular cells after drug treatment, and these findings may provide new targets for DKD therapy at the organelle level.https://doi.org/10.1186/s12967-025-06074-5snRNA-seqMitophagyMitochondrial homeostasis perturbationDiabetic kidney diseaseLosartan |
| spellingShingle | Zhen Zhu Guangxin Luan Song Wu Yiyi Song Shuang Shen Kaiyue Wu Shengnan Qian Weiping Jia Jun Yin Tao Ren Jianping Ye Li Wei Single-cell atlas reveals multi-faced responses of losartan on tubular mitochondria in diabetic kidney disease Journal of Translational Medicine snRNA-seq Mitophagy Mitochondrial homeostasis perturbation Diabetic kidney disease Losartan |
| title | Single-cell atlas reveals multi-faced responses of losartan on tubular mitochondria in diabetic kidney disease |
| title_full | Single-cell atlas reveals multi-faced responses of losartan on tubular mitochondria in diabetic kidney disease |
| title_fullStr | Single-cell atlas reveals multi-faced responses of losartan on tubular mitochondria in diabetic kidney disease |
| title_full_unstemmed | Single-cell atlas reveals multi-faced responses of losartan on tubular mitochondria in diabetic kidney disease |
| title_short | Single-cell atlas reveals multi-faced responses of losartan on tubular mitochondria in diabetic kidney disease |
| title_sort | single cell atlas reveals multi faced responses of losartan on tubular mitochondria in diabetic kidney disease |
| topic | snRNA-seq Mitophagy Mitochondrial homeostasis perturbation Diabetic kidney disease Losartan |
| url | https://doi.org/10.1186/s12967-025-06074-5 |
| work_keys_str_mv | AT zhenzhu singlecellatlasrevealsmultifacedresponsesoflosartanontubularmitochondriaindiabetickidneydisease AT guangxinluan singlecellatlasrevealsmultifacedresponsesoflosartanontubularmitochondriaindiabetickidneydisease AT songwu singlecellatlasrevealsmultifacedresponsesoflosartanontubularmitochondriaindiabetickidneydisease AT yiyisong singlecellatlasrevealsmultifacedresponsesoflosartanontubularmitochondriaindiabetickidneydisease AT shuangshen singlecellatlasrevealsmultifacedresponsesoflosartanontubularmitochondriaindiabetickidneydisease AT kaiyuewu singlecellatlasrevealsmultifacedresponsesoflosartanontubularmitochondriaindiabetickidneydisease AT shengnanqian singlecellatlasrevealsmultifacedresponsesoflosartanontubularmitochondriaindiabetickidneydisease AT weipingjia singlecellatlasrevealsmultifacedresponsesoflosartanontubularmitochondriaindiabetickidneydisease AT junyin singlecellatlasrevealsmultifacedresponsesoflosartanontubularmitochondriaindiabetickidneydisease AT taoren singlecellatlasrevealsmultifacedresponsesoflosartanontubularmitochondriaindiabetickidneydisease AT jianpingye singlecellatlasrevealsmultifacedresponsesoflosartanontubularmitochondriaindiabetickidneydisease AT liwei singlecellatlasrevealsmultifacedresponsesoflosartanontubularmitochondriaindiabetickidneydisease |