Intestinal fibrosis in aganglionic segment of Hirschsprung's disease revealed by single‐cell RNA sequencing
Abstract Background Hirschsprung's disease (HSCR) is a relatively common congenital disability. Accumulating extracellular matrix (ECM) prompts intestinal fibrosis remodelling in the aganglionic segments of HSCR. The contributions of various cellular subsets in the fibrogenesis of HSCR segments...
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| Format: | Article |
| Language: | English |
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Wiley
2023-02-01
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| Series: | Clinical and Translational Medicine |
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| Online Access: | https://doi.org/10.1002/ctm2.1193 |
| _version_ | 1797897189674975232 |
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| author | Shiwei He Junfeng Wang Yanlei Huang Fanyang Kong Ran Yang Yong Zhan Zifeng Li Chunjing Ye Lingdu Meng Yankang Ren Ying Zhou Gong Chen Zhen Shen Song Sun Shan Zheng Rui Dong |
| author_facet | Shiwei He Junfeng Wang Yanlei Huang Fanyang Kong Ran Yang Yong Zhan Zifeng Li Chunjing Ye Lingdu Meng Yankang Ren Ying Zhou Gong Chen Zhen Shen Song Sun Shan Zheng Rui Dong |
| author_sort | Shiwei He |
| collection | DOAJ |
| description | Abstract Background Hirschsprung's disease (HSCR) is a relatively common congenital disability. Accumulating extracellular matrix (ECM) prompts intestinal fibrosis remodelling in the aganglionic segments of HSCR. The contributions of various cellular subsets in the fibrogenesis of HSCR segments are poorly understood. Methods Single‐cell transcriptomics from 8 aganglionic segments and 5 normal segments of 7 HSCR subjects and 26 healthy segments of seven healthy donors were analysed. Fibrotic phenotype and alterations were explored using differential expression analysis and single‐cell trajectory analysis. Fibrosis‐related transcription factors were inferred through single‐cell regulatory network inference. Bulk transcriptomic data, proteomic data, immunohistochemistry (IHC) and real‐time polymerase chain reaction were used to validate the alterations in the HSCR intestine. Results Various collagen, fibronectin and laminin protein‐coding genes expression were up‐regulated in the stromal and glial cells of the HSCR intestine. The number of fibroblasts and myofibroblasts in the aganglionic segments increased, and more myofibroblasts were activated at an earlier stage in HSCR segments, which infers that there is an intestinal fibrosis phenotype in HSCR segments. The fibrotic regulators POSTN, ANXA1 and HSP70 were highly expressed in the ECM‐related cellular subsets in the transitional segments and aganglionic segments. The transcription factor regulatory network revealed that fibrosis‐related and megacolon‐related NR2F1 in the fibroblasts and glial subsets was up‐regulated in the aganglionic segment. Conclusions This work identifies intestinal fibrosis and related regulators in aganglionic segments of HSCR; hence, anti‐fibrotic therapy may be considered to prevent HSCR‐associated enterocolitis (HAEC), relieve intestinal stricture and improve cell therapy. |
| first_indexed | 2024-04-10T07:54:44Z |
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| id | doaj.art-e0cea7c961ee49c187172fa93f66f496 |
| institution | Directory Open Access Journal |
| issn | 2001-1326 |
| language | English |
| last_indexed | 2024-04-10T07:54:44Z |
| publishDate | 2023-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Clinical and Translational Medicine |
| spelling | doaj.art-e0cea7c961ee49c187172fa93f66f4962023-02-23T07:02:33ZengWileyClinical and Translational Medicine2001-13262023-02-01132n/an/a10.1002/ctm2.1193Intestinal fibrosis in aganglionic segment of Hirschsprung's disease revealed by single‐cell RNA sequencingShiwei He0Junfeng Wang1Yanlei Huang2Fanyang Kong3Ran Yang4Yong Zhan5Zifeng Li6Chunjing Ye7Lingdu Meng8Yankang Ren9Ying Zhou10Gong Chen11Zhen Shen12Song Sun13Shan Zheng14Rui Dong15Department of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaDepartment of Pediatric SurgeryShanghai Key Laboratory of Birth DefectChildren's Hospital of Fudan UniversityMinistry of Health ShanghaiChinaAbstract Background Hirschsprung's disease (HSCR) is a relatively common congenital disability. Accumulating extracellular matrix (ECM) prompts intestinal fibrosis remodelling in the aganglionic segments of HSCR. The contributions of various cellular subsets in the fibrogenesis of HSCR segments are poorly understood. Methods Single‐cell transcriptomics from 8 aganglionic segments and 5 normal segments of 7 HSCR subjects and 26 healthy segments of seven healthy donors were analysed. Fibrotic phenotype and alterations were explored using differential expression analysis and single‐cell trajectory analysis. Fibrosis‐related transcription factors were inferred through single‐cell regulatory network inference. Bulk transcriptomic data, proteomic data, immunohistochemistry (IHC) and real‐time polymerase chain reaction were used to validate the alterations in the HSCR intestine. Results Various collagen, fibronectin and laminin protein‐coding genes expression were up‐regulated in the stromal and glial cells of the HSCR intestine. The number of fibroblasts and myofibroblasts in the aganglionic segments increased, and more myofibroblasts were activated at an earlier stage in HSCR segments, which infers that there is an intestinal fibrosis phenotype in HSCR segments. The fibrotic regulators POSTN, ANXA1 and HSP70 were highly expressed in the ECM‐related cellular subsets in the transitional segments and aganglionic segments. The transcription factor regulatory network revealed that fibrosis‐related and megacolon‐related NR2F1 in the fibroblasts and glial subsets was up‐regulated in the aganglionic segment. Conclusions This work identifies intestinal fibrosis and related regulators in aganglionic segments of HSCR; hence, anti‐fibrotic therapy may be considered to prevent HSCR‐associated enterocolitis (HAEC), relieve intestinal stricture and improve cell therapy.https://doi.org/10.1002/ctm2.1193extracellular matrixHirschsprungintestinal fibrosissingle‐cell transcriptomics |
| spellingShingle | Shiwei He Junfeng Wang Yanlei Huang Fanyang Kong Ran Yang Yong Zhan Zifeng Li Chunjing Ye Lingdu Meng Yankang Ren Ying Zhou Gong Chen Zhen Shen Song Sun Shan Zheng Rui Dong Intestinal fibrosis in aganglionic segment of Hirschsprung's disease revealed by single‐cell RNA sequencing Clinical and Translational Medicine extracellular matrix Hirschsprung intestinal fibrosis single‐cell transcriptomics |
| title | Intestinal fibrosis in aganglionic segment of Hirschsprung's disease revealed by single‐cell RNA sequencing |
| title_full | Intestinal fibrosis in aganglionic segment of Hirschsprung's disease revealed by single‐cell RNA sequencing |
| title_fullStr | Intestinal fibrosis in aganglionic segment of Hirschsprung's disease revealed by single‐cell RNA sequencing |
| title_full_unstemmed | Intestinal fibrosis in aganglionic segment of Hirschsprung's disease revealed by single‐cell RNA sequencing |
| title_short | Intestinal fibrosis in aganglionic segment of Hirschsprung's disease revealed by single‐cell RNA sequencing |
| title_sort | intestinal fibrosis in aganglionic segment of hirschsprung s disease revealed by single cell rna sequencing |
| topic | extracellular matrix Hirschsprung intestinal fibrosis single‐cell transcriptomics |
| url | https://doi.org/10.1002/ctm2.1193 |
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