BACKGROUND: In Celiac disease (CD), cytoskeletal integrity of intestinal cells is disrupted by gliadin exposure. This study investigates the role of heat shock protein (Hsp)70 during cytoskeletal recovery in CD by assessing its induction and effects on junctional proteins. METHODS: Using an in-vitro model of CD, cytoskeletal injury and recovery was assessed in gliadin-exposed Caco-2 cells by measuring cellular distribution of ezrin, E-cadherin, and Hsp70 by differential centrifugation. Effects of Hsp70 were tested by an in-vitro repair assay, based on the incubation of injured or recovered cytoskeletal cellular fractions in noncytoskeletal supernatants containing low or high levels of Hsp70, or by transient transfection of Caco-2 cells with Hsp70. RESULTS: Cytoskeletal disruption of ezrin and E-cadherin was demonstrated in gliadin-exposed Caco-2 cells by their significant shift from the cytoskeletal pellet into the noncytoskeletal supernatant fraction. Recovery from gliadin exposure was associated with induction and cytoskeletal redistribution of Hsp70. The in-vitro repair assay delineated direct evidence for HSP-mediated repair by stabilization of junctional proteins by Hsp70. Overexpression of Hsp70 resulted in significantly increased cytoskeletal integrity. CONCLUSION: Our results establish an essential role of HSP-mediated cytoskeletal repair in Caco-2 cells during recovery from in-vitro gliadin exposure.
BACKGROUND: In Celiac disease (CD), cytoskeletal integrity of intestinal cells is disrupted by gliadin exposure. This study investigates the role of heat shock protein (Hsp)70 during cytoskeletal recovery in CD by assessing its induction and effects on junctional proteins. METHODS: Using an in-vitro model of CD, cytoskeletal injury and recovery was assessed in gliadin-exposed Caco-2 cells by measuring cellular distribution of ezrin, E-cadherin, and Hsp70 by differential centrifugation. Effects of Hsp70 were tested by an in-vitro repair assay, based on the incubation of injured or recovered cytoskeletal cellular fractions in noncytoskeletal supernatants containing low or high levels of Hsp70, or by transient transfection of Caco-2 cells with Hsp70. RESULTS: Cytoskeletal disruption of ezrin and E-cadherin was demonstrated in gliadin-exposed Caco-2 cells by their significant shift from the cytoskeletal pellet into the noncytoskeletal supernatant fraction. Recovery from gliadin exposure was associated with induction and cytoskeletal redistribution of Hsp70. The in-vitro repair assay delineated direct evidence for HSP-mediated repair by stabilization of junctional proteins by Hsp70. Overexpression of Hsp70 resulted in significantly increased cytoskeletal integrity. CONCLUSION: Our results establish an essential role of HSP-mediated cytoskeletal repair in Caco-2 cells during recovery from in-vitro gliadin exposure.
Authors: Sandro Drago; Ramzi El Asmar; Mariarosaria Di Pierro; Maria Grazia Clemente; Amit Tripathi; Anna Sapone; Manjusha Thakar; Giuseppe Iacono; Antonio Carroccio; Cinzia D'Agate; Tarcisio Not; Lucia Zampini; Carlo Catassi; Alessio Fasano Journal: Scand J Gastroenterol Date: 2006-04 Impact factor: 2.423
Authors: Michael Schumann; Sarah Kamel; Marie-Luise Pahlitzsch; Lydia Lebenheim; Claudia May; Michael Krauss; Michael Hummel; Severin Daum; Michael Fromm; Jörg-Dieter Schulzke Journal: Ann N Y Acad Sci Date: 2012-07 Impact factor: 5.691
Authors: Bettina Bidmon; Michaela Endemann; Thomas Müller; Klaus Arbeiter; Kurt Herkner; Christoph Aufricht Journal: Kidney Int Date: 2002-11 Impact factor: 10.612
Authors: Mark J Ropeleski; Jun Tang; Margaret M Walsh-Reitz; Mark W Musch; Eugene B Chang Journal: Gastroenterology Date: 2003-05 Impact factor: 22.682
Authors: M G Clemente; S De Virgiliis; J S Kang; R Macatagney; M P Musu; M R Di Pierro; S Drago; M Congia; A Fasano Journal: Gut Date: 2003-02 Impact factor: 23.059
Authors: Ruben M L Colunga Biancatelli; Pavel Solopov; Christiana Dimitropoulou; Betsy Gregory; Tierney Day; John D Catravas Journal: Cells Date: 2022-03-19 Impact factor: 6.600