Role of the intestinal epithelial barrier in IBD

Ulcerative colitis (UC) and Crohn’s Disease (CD) are chronic inflammatory disorders whose etiology is still unknown. Increasing evidence, however, seems to demonstrate that primary changes in the intestinal epithelial barrier could play a role in the onset and perpetuation (paper Nuria) of these diseases.

 

In order to more deeply investigate the contribution of the epithelium in IBD, we have recently set up in our laboratory a protocol for long-term in vitro expansion of human epithelial stem cells (hESCs) as 3D spheroid cultures from freshly isolated crypts. This protocol has been optimized using clinical samples (both surgical and biopsy specimens) from healthy subjects and IBD patients.

Expansion of hESCs

INTESTINAL CRYPTS

INTESTINAL SPHEROIDS

We are currently exploiting this culture system to explore whether primary defects are present in the epithelial stem cell compartment of IBD patients by investigating the presence of expression profile differences in the spheroid cultures derived from IBD samples.

 

Graphical view of human colon mucosa and epithelial crypt. Left side shows a graphical section of the normal colon wall where the primary layers are can bee seen. Right side depicts epithelial crypt cell types and gradients of BMP (bone morphogenic protein), Notch and Wnt signaling (drawing by Rut Mora, PhD).

 

In parallel, we are investigating the potential of the hESC culture system as a cell-based tool for the treatment of those IBD patients refractory to the standard therapeutic protocols. Our approach is currently focused on optimizing the engraftment and regeneration capacity of rat and human spheroids in animal models of intestinal inflammation (both rats and mice).

Immunofluorescent staining of differentiated organoids in culture.

This project is directed by Azucena Salas. Isabella Dotti is involved in the generation of organoid cultures. Nuría Planell is in charge of the microarray analysis. Elena Ferrer is involved in all of the transplantation experiments. We are actively collaborating with Dr. Eduard Batlle’s group at the Institut de Recerca Biomèdica (IRB Barcelo na), which helped us in the establishment of the human spheroid culture.

Funding:

- Fundación Marató TV3. PI: Azucena Salas and Eduard Batlle.

- R+D National Fund from Spanish Minestry of Economy and Innovation (I+D BFU2012-35999/BFI); PI: Azucena Salas.

- Leona and Harry Helmsley Trust; PI: Azucena Salas

Publications:

 

Potential Use of Human Stem Cell-Derived Intestinal Organoids to Study Inflammatory Bowel Diseases.

Dotti I, Salas A.

Inflamm Bowel Dis. 2018 Nov 29;24(12):2501-2509.

doi: 10.1093/ibd/izy275.

Alterations in the epithelial stem cell compartment could contribute to permanent changes in the mucosa of patients with ulcerative colitis.

Dotti I, Mora-Buch R, Ferrer-Picón E, Planell N, Jung P, Masamunt MC, Leal RF, Martín de Carpi J, Llach J, Ordás I, Batlle E, Panés J, Salas A.

Gut. 2017 Dec;66(12):2069-2079.

doi: 10.1136/gutjnl-2016-312609. Epub 2016 Nov 1.

 

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