Altered epithelial barrier functions in the colon of patients with spina bifida

Charlène Brochard, Guillaume Bouguen, Raphael Olivier, Tony Durand, Sébastien Henno, Benoît Peyronnet, Mael Pagenault, Chloé Lefèvre, Gaëlle Boudry, Mikael Croyal, Alain Fautrel, Maxime Esvan, Alain Ropert, Anne Dariel, Laurent Siproudhis, Michel Neunlist, Charlène Brochard, Guillaume Bouguen, Raphael Olivier, Tony Durand, Sébastien Henno, Benoît Peyronnet, Mael Pagenault, Chloé Lefèvre, Gaëlle Boudry, Mikael Croyal, Alain Fautrel, Maxime Esvan, Alain Ropert, Anne Dariel, Laurent Siproudhis, Michel Neunlist

Abstract

Our objectives were to better characterize the colorectal function of patients with Spina Bifida (SB). Patients with SB and healthy volunteers (HVs) completed prospectively a standardized questionnaire, clinical evaluation, rectal barostat, colonoscopy with biopsies and faecal collection. The data from 36 adults with SB (age: 38.8 [34.1-47.2]) were compared with those of 16 HVs (age: 39.0 [31.0-46.5]). Compared to HVs, rectal compliance was lower in patients with SB (p = 0.01), whereas rectal tone was higher (p = 0.0015). Ex vivo paracellular permeability was increased in patients with SB (p = 0.0008) and inversely correlated with rectal compliance (r = - 0.563, p = 0.002). The expression of key tight junction proteins and inflammatory markers was comparable between SB and HVs, except for an increase in Claudin-1 immunoreactivity (p = 0.04) in SB compared to HVs. TGFβ1 and GDNF mRNAs were expressed at higher levels in patients with SB (p = 0.02 and p = 0.008). The levels of acetate, propionate and butyrate in faecal samples were reduced (p = 0.04, p = 0.01, and p = 0.02, respectively). Our findings provide evidence that anorectal and epithelial functions are altered in patients with SB. The alterations in these key functions might represent new therapeutic targets, in particular using microbiota-derived approaches.Clinical Trials: NCT02440984 and NCT03054415.

Conflict of interest statement

The authors declare no competing interests.

© 2022. The Author(s).

Figures

Figure 1
Figure 1
Rectal adaptation to isobaric distension. (A) Increasing the pressure of distension increased the recorded initial rectal volume (pressure effect, p < 0.0001). Compared with the HVs, the recorded initial rectal volumes (rectal compliance) were significantly lower in the patients with SB (group effect, p = 0.01; interaction group pression, p = 0.04). (B) Increasing the pressure of distension increased the recorded maximal rectal volumes (pressure effect, p < 0.0001). (C) Increasing pressure of distension decreased the rectal tone (maximal volume–initial volume) up to the treshold of 21 mmHg (pressure effect, p < 0.0001) for both groups. Compared with the HVs, rectal tone was significantly higher in the patients with SB up to the treshold of 21 mmHg (group effect, p = 0.08; interaction group pression; p = 0.001). *Patients with Spina Bifida; O Healthy volunteers.
Figure 2
Figure 2
(A) and (B) Comparison of para- and transcellular permeability in patients with Spina Bifida and healthy volunteers (Hvs). (A) The paracellular permeability was significantly increased in patients with SB (p = 0008). (B) For the evaluation of transcellular permeability, no significant changes were observed between the two groups (p = 0.16). (C,D) The expression levels of claudin 1 and ZO1 assessed in Western Blot were comparable between the two groups. (D) The average claudin 1 area/crypt tended to be lower in patients with SB (p = 0.08). (E) The average ZO1 area /crypt were comparable between the two groups.
Figure 3
Figure 3
(A) A first macroscopic image of the whole biopsy was first obtained using a wide-field camera in order to screen for the regions of interest within the biopsy. Static FF-OCT images of the region of interest were then obtained at 2 depths. (B) Two images for each subject were assessed. (C) Image of biopsy of Hv stained with Sirius red. (D) Image of biopsy of SB stained with Sirius red. (E) The percentage of collagen per area were significantly lower in patients with SB (p = 0.0003). (F) The percentage of collagen per area after staining with red sirius was significantly lower in patients with SB (p = 0.0075). (G) The mRNA expression of TGF beta was significantly higher in patients with SB compared with Hvs (p = 0.0176). (H) The mRNA expression of MMP2 was significantly higher in patients with SB compared with Hvs (p = 0.0121) and tended to be correlated with the paracellular permeability (p = 0.0516) (I). (J) The TIMP1/MMP1 balance was significantly lower in patients with Spina Bifida (p = 0.0348).
Figure 4
Figure 4
The expression levels of acetate, propionate and butyrate in fecal samples were significantly lower by 33, 54 and 53% in patients with SB compared with HVs (p = 0.02, p = 0.01, p = 0.02, respectively).

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