DCLK1 isoforms and aberrant Notch signaling in the regulation of human and murine colitis
Abstract Alternative promoter usage generates long and short isoforms (DCLK1-L and DCLK1-S) of doublecortin-like kinase-1 (DCLK1). Tight control of Notch signaling is important to prevent and restitute inflammation in the intestine. Our aim was to investigate whether Notch1–DCLK1 axis regulates the...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Publishing Group
2021-06-01
|
| Series: | Cell Death Discovery |
| Online Access: | https://doi.org/10.1038/s41420-021-00526-9 |
| _version_ | 1818738158226898944 |
|---|---|
| author | Badal C. Roy Ishfaq Ahmed Jason Stubbs Jun Zhang Thomas Attard Seth Septer Danny Welch Shrikant Anant Venkatesh Sampath Shahid Umar |
| author_facet | Badal C. Roy Ishfaq Ahmed Jason Stubbs Jun Zhang Thomas Attard Seth Septer Danny Welch Shrikant Anant Venkatesh Sampath Shahid Umar |
| author_sort | Badal C. Roy |
| collection | DOAJ |
| description | Abstract Alternative promoter usage generates long and short isoforms (DCLK1-L and DCLK1-S) of doublecortin-like kinase-1 (DCLK1). Tight control of Notch signaling is important to prevent and restitute inflammation in the intestine. Our aim was to investigate whether Notch1–DCLK1 axis regulates the mucosal immune responses to infection and whether this is phenocopied in human models of colitis. In the FFPE (formalin-fixed paraffin-embedded) sections prepared from the colons of ulcerative colitis (UC) and immune-mediated colitis (IRAEC) patients, expression of DCLK1 isoforms correlated positively with Notch1 and negatively with a transcriptional repressor, FoxD3 (Forkhead Box D3). DCLK1 protein staining in these sections was predominantly sub-epithelial (stromal) wherein DCLK1 co-localized with NICD, CD68, CD11c, and neutrophil elastase (NE). NE also co-stained with Citrullinated-H3 indicating the presence of neutrophil extracellular traps. In human neutrophils, elevated levels of DCLK1-S, CXCL-10, Ly6G, MPO, NE, and Notch1/2 in LPS-treated cells were inhibited when LPS was added in conjunction with Notch blocker dibenzazepine (DBZ; LPS + DBZ group). In CR-infected Rag1 −/− mice, higher levels of DCLK1 in the colonic crypts were inhibited when mice received DBZ for 10 days coincident with significant dysbiosis, barrier disruption, and colitis. Concurrently, DCLK1 immunoreactivity shifted toward the stroma in CR + DBZ mice with predominance of DCLK1-S that coincided with higher Notch1 levels. Upon antibiotic treatment, partial restoration of crypt DCLK1, reduction in MPO activity, and increased survival followed. When intestinal epithelial cell-specific Dclk1-knockout (Dclk1 ΔIEC ) or Dclk1 ΔIEC ;Rag1 −/− double knockout (DKO) mice were infected with CR and given a single dose of DBZ, they developed barrier defect and severe colitis with higher levels of stromal DCLK1-S, Ly6G, NE, and Notch1. We therefore propose that, by regulating the mucosal immune responses, the Notch–DCLK1 axis may be integral to the development of murine or human colitis. |
| first_indexed | 2024-12-18T01:04:29Z |
| format | Article |
| id | doaj.art-d7436833a6ef4b55967dec6aa603e365 |
| institution | Directory Open Access Journal |
| issn | 2058-7716 |
| language | English |
| last_indexed | 2024-12-18T01:04:29Z |
| publishDate | 2021-06-01 |
| publisher | Nature Publishing Group |
| record_format | Article |
| series | Cell Death Discovery |
| spelling | doaj.art-d7436833a6ef4b55967dec6aa603e3652022-12-21T21:26:17ZengNature Publishing GroupCell Death Discovery2058-77162021-06-017111410.1038/s41420-021-00526-9DCLK1 isoforms and aberrant Notch signaling in the regulation of human and murine colitisBadal C. Roy0Ishfaq Ahmed1Jason Stubbs2Jun Zhang3Thomas Attard4Seth Septer5Danny Welch6Shrikant Anant7Venkatesh Sampath8Shahid Umar9Department of Surgery, University of Kansas Medical CenterDepartment of Surgery, University of Kansas Medical CenterDepartment of Internal Medicine, University of Kansas Medical CenterDepartment of Internal Medicine, University of Kansas Medical CenterChildren’s Mercy HospitalChildren’s HospitalDepartment of Cancer Biology, University of Kansas Medical CenterDepartment of Cancer Biology, University of Kansas Medical CenterChildren’s Mercy HospitalDepartment of Surgery, University of Kansas Medical CenterAbstract Alternative promoter usage generates long and short isoforms (DCLK1-L and DCLK1-S) of doublecortin-like kinase-1 (DCLK1). Tight control of Notch signaling is important to prevent and restitute inflammation in the intestine. Our aim was to investigate whether Notch1–DCLK1 axis regulates the mucosal immune responses to infection and whether this is phenocopied in human models of colitis. In the FFPE (formalin-fixed paraffin-embedded) sections prepared from the colons of ulcerative colitis (UC) and immune-mediated colitis (IRAEC) patients, expression of DCLK1 isoforms correlated positively with Notch1 and negatively with a transcriptional repressor, FoxD3 (Forkhead Box D3). DCLK1 protein staining in these sections was predominantly sub-epithelial (stromal) wherein DCLK1 co-localized with NICD, CD68, CD11c, and neutrophil elastase (NE). NE also co-stained with Citrullinated-H3 indicating the presence of neutrophil extracellular traps. In human neutrophils, elevated levels of DCLK1-S, CXCL-10, Ly6G, MPO, NE, and Notch1/2 in LPS-treated cells were inhibited when LPS was added in conjunction with Notch blocker dibenzazepine (DBZ; LPS + DBZ group). In CR-infected Rag1 −/− mice, higher levels of DCLK1 in the colonic crypts were inhibited when mice received DBZ for 10 days coincident with significant dysbiosis, barrier disruption, and colitis. Concurrently, DCLK1 immunoreactivity shifted toward the stroma in CR + DBZ mice with predominance of DCLK1-S that coincided with higher Notch1 levels. Upon antibiotic treatment, partial restoration of crypt DCLK1, reduction in MPO activity, and increased survival followed. When intestinal epithelial cell-specific Dclk1-knockout (Dclk1 ΔIEC ) or Dclk1 ΔIEC ;Rag1 −/− double knockout (DKO) mice were infected with CR and given a single dose of DBZ, they developed barrier defect and severe colitis with higher levels of stromal DCLK1-S, Ly6G, NE, and Notch1. We therefore propose that, by regulating the mucosal immune responses, the Notch–DCLK1 axis may be integral to the development of murine or human colitis.https://doi.org/10.1038/s41420-021-00526-9 |
| spellingShingle | Badal C. Roy Ishfaq Ahmed Jason Stubbs Jun Zhang Thomas Attard Seth Septer Danny Welch Shrikant Anant Venkatesh Sampath Shahid Umar DCLK1 isoforms and aberrant Notch signaling in the regulation of human and murine colitis Cell Death Discovery |
| title | DCLK1 isoforms and aberrant Notch signaling in the regulation of human and murine colitis |
| title_full | DCLK1 isoforms and aberrant Notch signaling in the regulation of human and murine colitis |
| title_fullStr | DCLK1 isoforms and aberrant Notch signaling in the regulation of human and murine colitis |
| title_full_unstemmed | DCLK1 isoforms and aberrant Notch signaling in the regulation of human and murine colitis |
| title_short | DCLK1 isoforms and aberrant Notch signaling in the regulation of human and murine colitis |
| title_sort | dclk1 isoforms and aberrant notch signaling in the regulation of human and murine colitis |
| url | https://doi.org/10.1038/s41420-021-00526-9 |
| work_keys_str_mv | AT badalcroy dclk1isoformsandaberrantnotchsignalingintheregulationofhumanandmurinecolitis AT ishfaqahmed dclk1isoformsandaberrantnotchsignalingintheregulationofhumanandmurinecolitis AT jasonstubbs dclk1isoformsandaberrantnotchsignalingintheregulationofhumanandmurinecolitis AT junzhang dclk1isoformsandaberrantnotchsignalingintheregulationofhumanandmurinecolitis AT thomasattard dclk1isoformsandaberrantnotchsignalingintheregulationofhumanandmurinecolitis AT sethsepter dclk1isoformsandaberrantnotchsignalingintheregulationofhumanandmurinecolitis AT dannywelch dclk1isoformsandaberrantnotchsignalingintheregulationofhumanandmurinecolitis AT shrikantanant dclk1isoformsandaberrantnotchsignalingintheregulationofhumanandmurinecolitis AT venkateshsampath dclk1isoformsandaberrantnotchsignalingintheregulationofhumanandmurinecolitis AT shahidumar dclk1isoformsandaberrantnotchsignalingintheregulationofhumanandmurinecolitis |