Wei-Ting Kuo1, Li Zuo2, Matthew A Odenwald3, Shariq Madha4, Gurminder Singh1, Christine B Gurniak5, Clara Abraham6, Jerrold R Turner7. 1. Laboratory of Mucosal Barrier Pathobiology, Department of Pathology, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts. 2. Laboratory of Mucosal Barrier Pathobiology, Department of Pathology, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts; Anhui Medical University, Hefei, Anhui, China. 3. Department of Pathology, The University of Chicago, Chicago, Illinois. 4. Department of Medical Oncology and Center for Functional Cancer Epigenetics, Dana-Farber Cancer Institute, Boston, Massachusetts. 5. Institute of Genetics, University of Bonn, Bonn, Germany. 6. Department of Internal Medicine, Yale University, New Haven, Connecticut. 7. Laboratory of Mucosal Barrier Pathobiology, Department of Pathology, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts; Department of Pathology, The University of Chicago, Chicago, Illinois. Electronic address: jrturner@bwh.harvard.edu.
Abstract
BACKGROUNDS & AIMS: Increased permeability is implicated in the pathogenesis of intestinal disease. In vitro and in vivo studies have linked down-regulation of the scaffolding protein ZO-1, encoded by the TJP1 gene, to increased tight junction permeability. This has not, however, been tested in vivo. Here, we assessed the contributions of ZO-1 to in vivo epithelial barrier function and mucosal homeostasis. METHODS: Public Gene Expression Omnibus data sets and biopsy specimens from patients with inflammatory bowel disease (IBD) and healthy control individuals were analyzed. Tjp1f/f;vil-CreTg mice with intestinal epithelial-specific ZO-1 knockout (ZO-1KO.IEC) mice and Tjp1f/f mice littermates without Cre expression were studied using chemical and immune-mediated models of disease as well as colonic stem cell cultures. RESULTS: ZO-1 transcript and protein expression were reduced in biopsy specimens from patients with IBD. Despite mildly increased intestinal permeability, ZO-1KO.IEC mice were healthy and did not develop spontaneous disease. ZO-1KO.IEC mice were, however, hypersensitive to mucosal insults and displayed defective repair. Furthermore, ZO-1-deficient colonic epithelia failed to up-regulate proliferation in response to damage in vivo or Wnt signaling in vitro. ZO-1 was associated with centrioles in interphase cells and mitotic spindle poles during division. In the absence of ZO-1, mitotic spindles failed to correctly orient, resulting in mitotic catastrophe and abortive proliferation. ZO-1 is, therefore, critical for up-regulation of epithelial proliferation and successful completion of mitosis. CONCLUSIONS: ZO-1 makes critical, tight junction-independent contributions to Wnt signaling and mitotic spindle orientation. As a result, ZO-1 is essential for mucosal repair. We speculate that ZO-1 down-regulation may be one cause of ineffective mucosal healing in patients with IBD.
BACKGROUNDS & AIMS: Increased permeability is implicated in the pathogenesis of intestinal disease. In vitro and in vivo studies have linked down-regulation of the scaffolding protein ZO-1, encoded by the TJP1 gene, to increased tight junction permeability. This has not, however, been tested in vivo. Here, we assessed the contributions of ZO-1 to in vivo epithelial barrier function and mucosal homeostasis. METHODS: Public Gene Expression Omnibus data sets and biopsy specimens from patients with inflammatory bowel disease (IBD) and healthy control individuals were analyzed. Tjp1f/f;vil-CreTg mice with intestinal epithelial-specific ZO-1 knockout (ZO-1KO.IEC) mice and Tjp1f/f mice littermates without Cre expression were studied using chemical and immune-mediated models of disease as well as colonic stem cell cultures. RESULTS: ZO-1 transcript and protein expression were reduced in biopsy specimens from patients with IBD. Despite mildly increased intestinal permeability, ZO-1KO.IEC mice were healthy and did not develop spontaneous disease. ZO-1KO.IEC mice were, however, hypersensitive to mucosal insults and displayed defective repair. Furthermore, ZO-1-deficient colonic epithelia failed to up-regulate proliferation in response to damage in vivo or Wnt signaling in vitro. ZO-1 was associated with centrioles in interphase cells and mitotic spindle poles during division. In the absence of ZO-1, mitotic spindles failed to correctly orient, resulting in mitotic catastrophe and abortive proliferation. ZO-1 is, therefore, critical for up-regulation of epithelial proliferation and successful completion of mitosis. CONCLUSIONS: ZO-1 makes critical, tight junction-independent contributions to Wnt signaling and mitotic spindle orientation. As a result, ZO-1 is essential for mucosal repair. We speculate that ZO-1 down-regulation may be one cause of ineffective mucosal healing in patients with IBD.
Authors: Wei-Ting Kuo; Le Shen; Li Zuo; Nitesh Shashikanth; Ma Lora Drizella M Ong; Licheng Wu; Juanmin Zha; Karen L Edelblum; Yitang Wang; Yingmin Wang; Steven P Nilsen; Jerrold R Turner Journal: Gastroenterology Date: 2019-08-08 Impact factor: 22.682
Authors: Matthew A Odenwald; Wangsun Choi; Aaron Buckley; Nitesh Shashikanth; Nora E Joseph; Yitang Wang; Michael H Warren; Mary M Buschmann; Roman Pavlyuk; Jeffrey Hildebrand; Ben Margolis; Alan S Fanning; Jerrold R Turner Journal: J Cell Sci Date: 2016-10-21 Impact factor: 5.285
Authors: Williams Turpin; Sun-Ho Lee; Juan Antonio Raygoza Garay; Karen L Madsen; Jonathan B Meddings; Larbi Bedrani; Namita Power; Osvaldo Espin-Garcia; Wei Xu; Michelle I Smith; Anne M Griffiths; Paul Moayyedi; Dan Turner; Ernest G Seidman; A Hillary Steinhart; John K Marshall; Kevan Jacobson; David Mack; Hien Huynh; Charles N Bernstein; Andrew D Paterson; Kenneth Croitoru Journal: Gastroenterology Date: 2020-08-10 Impact factor: 22.682