Clinical trial NCT05916274: Exploration of the Activity of DNA Located Outside of Cellular Nucleus to Amplify Inflammation in Inflammatory Bowel Disease in Children Through Biological Pathway Cyclic GMP-AMP Synthase (cGAS) - Stimulator of Interferon Genes (STING)
Official title: Pro-inflammatory Role of Extracellular DNA in Inflammatory Bowel Disease in Children: Study of the cGAS-STING Pathway
Study type: INTERVENTIONAL
Overall status: COMPLETED
Phase: NA
Conditions: Inflammatory Bowel Diseases
Crohn Disease
Ulcerative Colitis
Keywords: Inflammatory Bowel disease
Crohn disease
Ulcerative colitis
cell-free DNA
cGAS
STING
Child
intestinal inflammation
Lead sponsor: Centre Hospitalier Régional d'Orléans
Sponsor class: OTHER
First posted: 2023-06-23
Enrollment: 40
Sex: ALL
Minimum age (years): 6
Maximum age (years): 17
Standard ages: CHILD
Accepts healthy volunteers: no
Allocation: NA
Intervention model: SINGLE_GROUP
Primary purpose: HEALTH_SERVICES_RESEARCH

Brief summary: Frequency of Inflammatory Bowel Diseases in children (IBD)-Crohn's disease (CD), Ulcerative colitis (UC) is constantly increasing. Pediatric-onset IBD represent a different nosological entity (from adult IBD) because of their major inflammatory activity, their significant anatomical extent and their stenotic and/or fistulizing character sometimes from diagnosis. Intestinal lesions are due to dysregulation of the intestinal immune system but the cause is unknown. The investigators hypothesize that extranuclear DNA participates in the amplification of the inflammatory response at the intestinal and blood levels during pediatric IBD through the cGAS-STING pathway. The investigators will analyse blood and fecal samples, and colonic biopsies issued from ill children and control participants on age of 6 to 17 years. The investigators think that this study will provide a better understanding of the mechanisms involved in pediatric IBD, assess the place of the cGAS-STING pathway, identify potential biomarkers of pediatric IBD and new potential therapeutic targets based in particular on the inhibition of the cGAS-STING pathway.

Detailed description: Inflammatory Bowel Diseases in children (IBD)-Crohn's disease (CD), ulcerative colitis (UC) are severe pathology that can affect the entire digestive tract. Their annual incidence is however constantly increasing.

IBD are complex multifactorial pathologies whose cause is still unknown today. IBD occurs on a predisposing genetic background in the presence of exogenous factors and alteration of the intestinal microbiota. Intestinal lesions are due to dysregulation of the intestinal immune system with increased secretion of pro-inflammatory cytokines at the expense of anti-inflammatory cytokines.

Pediatric-onset IBD represent a different nosological entity (from adult IBD) because of their major inflammatory activity, their significant anatomical extent and their stenotic and/or fistulizing character sometimes from diagnosis. Their impact is not only individual (growth retardation, puberty delay, psychological disorders) but also family/parental, school and social. These particularities justify that biomedical research focuses on it in a more specific way.

Extracellular and extranuclear DNA (enDNA) play a major role in innate immunity by stimulating pro-inflammatory responses and activating type I interferon production. The pro-inflammatory action of enDNA is mediated by enzyme cGAS, protein STING, toll-like receptor 9 (TLR9), and the inflammasome complex NLRP3.

The investigators hypothesize that enDNA participates in the amplification of the inflammatory response at the intestinal and blood levels during pediatric IBD through the cGAS-STING pathway. They also hypothesize that there are links between the cGAS-STING pathway and other pathways involved in pediatric IBD such as NOD2 and Autophagy. The investigators will analyse blood and fecal samples, and colonic biopsies issued from ill children and controls on age of 6 to 17 years. The investigators think that this study will provide a better understanding of the mechanisms involved in pediatric IBD, assess the place of the cGAS-STING pathway, identify potential biomarkers of pediatric IBD and new potential therapeutic targets based in particular on the inhibition of the cGAS-STING pathway.

Eligibility criteria: Inclusion Criteria:

* Participants aged from 6 years inclusive to 17 years inclusive
* Boys and girls
* Presenting an IBD or suspicion of IBD
* Requiring a colonoscopy for diagnosis or follow-up or other reason (abdominal pain, diarrhoea, rectal bleeding, weight loss) not confirming the diagnosis of Crohn's disease or Ulcerative Colitis
* Active IBD if:

  * CD: PCDAI score \>5 and CRP\>20mg/L and faecal calprotectin\> 400 µg/g
  * UC: PUCAI score\>10 and faecal calprotectin\>250 µg/g
* IBD in remission if:

  * CD: PCDAI score\<5 and CRP\<20mg/L and faecal calprotectin\<400 µg/g
  * UC: PUCAI score \<10 and faecal calprotectin \< 250 µg/g
* Patients and their parents who gave their consent to participate in the study

Exclusion Criteria:

* Refusal of the participant and/or one of his two parents
* Body weight less than or equal to 20 kg
* Blood hemoglobin level less than or equal to 9 g/dl
* Refusal or contraindication to general anesthesia
* Co-existing severe chronic pathology and/or treatment that could interfere with the results of the study; example: trisomy 21, treatment with growth hormone etc.
* Protected person (under guardianship or curatorship)
* Person under legal protection
* Person not affiliated to a social security scheme
* Pregnant or breastfeeding woman

Primary outcomes:
measure: Amount of mRNA specific for colonic cGAS
timeFrame: Baseline
Secondary outcomes:
measure: quantitative difference of amount of circulating mtDNA
timeFrame: Baseline
measure: Cytokine response
timeFrame: Baseline
measure: inflammatory / dysimmune response of cytokines
timeFrame: Baseline
measure: Difference of DNA methylation by Methyl-Seq between the 3 groups
timeFrame: Baseline
measure: Activation (phosphorylation) of the components of the cGAS-STING pathway
timeFrame: Baseline
measure: plasma DNase activity
timeFrame: baseline
measure: differences in microbial distribution
timeFrame: Baseline
measure: quantity (number of "reads") of colonic STING-specific mRNA
timeFrame: Baseline
measure: extracellular DNA by qPCR
timeFrame: Baseline
measure: Amount of STING in the cytoplasm
timeFrame: Baseline
measure: Amount of cGAS in the cytoplasm
timeFrame: Baseline
measure: Amount of nuclear DNA in teh cytoplasm
timeFrame: Baseline
measure: Amount of mitochondrial DNA in the cytoplasm
timeFrame: Baseline
measure: Amount of colonic DNase activity
timeFrame: Baseline
Other outcomes:
Locations:
city: Orléans
country: France
facility: CHR Orléans
Study officials:
affiliation: CHR d'Orleans
name: Georges DIMITROV, MD
role: PRINCIPAL_INVESTIGATOR
References:
citation: Ahn J, Son S, Oliveira SC, Barber GN. STING-Dependent Signaling Underlies IL-10 Controlled Inflammatory Colitis. Cell Rep. 2017 Dec 26;21(13):3873-3884. doi: 10.1016/j.celrep.2017.11.101.
citation: Canesso MCC, Lemos L, Neves TC, Marim FM, Castro TBR, Veloso ES, Queiroz CP, Ahn J, Santiago HC, Martins FS, Alves-Silva J, Ferreira E, Cara DC, Vieira AT, Barber GN, Oliveira SC, Faria AMC. The cytosolic sensor STING is required for intestinal homeostasis and control of inflammation. Mucosal Immunol. 2018 May;11(3):820-834. doi: 10.1038/mi.2017.88. Epub 2017 Dec 20.
citation: Martin GR, Blomquist CM, Henare KL, Jirik FR. Stimulator of interferon genes (STING) activation exacerbates experimental colitis in mice. Sci Rep. 2019 Oct 3;9(1):14281. doi: 10.1038/s41598-019-50656-5.
citation: Boyapati RK, Dorward DA, Tamborska A, Kalla R, Ventham NT, Doherty MK, Whitfield PD, Gray M, Loane J, Rossi AG, Satsangi J, Ho GT. Mitochondrial DNA Is a Pro-Inflammatory Damage-Associated Molecular Pattern Released During Active IBD. Inflamm Bowel Dis. 2018 Sep 15;24(10):2113-2122. doi: 10.1093/ibd/izy095.
citation: Vrablicova Z, Tomova K, Tothova L, Babickova J, Gromova B, Konecna B, Liptak R, Hlavaty T, Gardlik R. Nuclear and Mitochondrial Circulating Cell-Free DNA Is Increased in Patients With Inflammatory Bowel Disease in Clinical Remission. Front Med (Lausanne). 2020 Dec 14;7:593316. doi: 10.3389/fmed.2020.593316. eCollection 2020.
citation: Khan S, Mentrup HL, Novak EA, Siow VS, Wang Q, Crawford EC, Schneider C, Comerford TE 4th, Firek B, Rogers MB, Loughran P, Morowitz MJ, Mollen KP. Cyclic GMP-AMP synthase contributes to epithelial homeostasis in intestinal inflammation via Beclin-1-mediated autophagy. FASEB J. 2022 May;36(5):e22282. doi: 10.1096/fj.202200138R.
citation: Zhao F, Zheng T, Gong W, Wu J, Xie H, Li W, Zhang R, Liu P, Liu J, Wu X, Zhao Y, Ren J. Extracellular vesicles package dsDNA to aggravate Crohn's disease by activating the STING pathway. Cell Death Dis. 2021 Aug 27;12(9):815. doi: 10.1038/s41419-021-04101-z.
citation: Chen C, Zhang Y, Tao M, Zhao X, Feng Q, Fei X, Fu Y. Atrial Natriuretic Peptide Attenuates Colitis via Inhibition of the cGAS-STING Pathway in Colonic Epithelial Cells. Int J Biol Sci. 2022 Feb 7;18(4):1737-1754. doi: 10.7150/ijbs.67356. eCollection 2022.