Development and Use of a Tissue and Human Enteroid Biorepository to Study the Pathophysiology of NEC

• Conditions: Enterocolitis, Necrotizing; Prematurity; Gastrointestinal Disease
• Interventions: Other: Organoids creation

• Registration Number: NCT04549727
• Lead Sponsor: Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico

Brief Summary

Despite a greater understanding of NEC physiopathology, modest progress has been done in terms of intervention and prevention of the disease over the past three decades, being the mortality rate unchanged.

Investigators intend to leverage our knowledge and technical expertise developed with fetal enteroids to further investigate the processes leading to NEC by deriving and performing functional studies on human intestinal enteroids generated from intestinal resection for therapeutic reasons in NEC and non-NEC patients

1. Generate a tissue biorepository composed of: enteroids and other lamina propria cells

2. Comparative studies of the gene expression profile of tissue, epithelial enteroids and underlying lamina propria of NEC, non-NEC, hypoxic and non-hypoxic infants

3. In vitro functional studies for the evaluation of critical factors in NEC pathophysiology

4. In vitro functional studies to identify the activation of processes leading to intestinal epithelium necroptosis and/or apoptosis in bacteria challenged and hypoxic conditions

5. Correlative studies of the impact of perinatal variables on the intestinal barrier functionality at baseline and challenged with pathogens

6. In vitro comparison of the intestinal barrier functionality in infants complicated by condition of prenatal hypoxia versus non hypoxic infants

7. Validation the NEC enteroids as an in vitro model for the identification of treatments and prevention of NEC

Detailed Description

Not available

Study Timeline

• Status Verified: 2020-09
• Study First Submitted: 2020-09-09
• Study First Submitted QC: 2020-09-09
• Last Update Submitted: 2020-09-15
• Study First Posted: 2020-09-16
• Last Update Posted: 2020-09-17
• Study Start: 2020-11-01
• Primary Completion: 2022-02-17
• Study Completion: 2022-02-17

Recruitment & Eligibility

• Status: UNKNOWN
• Sex: All
• Target Recruitment: 18

Inclusion Criteria

• Infants, with a postconceptional age below 44 weeks of gestation, undergoing a clinically indicated intervention of partial intestinal resection while hospitalized at the NICU of the Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico.

Exclusion Criteria

• Infants presenting a devastating damage of the intestine

Study & Design

• Study Type: OBSERVATIONAL
• Study Design: Not specified

Arm && Interventions

Group	Intervention	Description
infants with surgical NEC	Organoids creation	Infants who undergo surgery for NEC disease
Infants with GI surgical diseases other than NEC	Organoids creation	Infants who undergo surgery for other GI diseases than NEC

Primary Outcome Measures

Name	Time	Method
Correlative studies of the impact of perinatal variables	2 years	Assess how the perinatal features (expression of the neonatal phenotype, as IUGR, chorionamnionitis, perinatal hypoxia) on the intestinal barrier functionality at baseline and challenged with pathogens
functional studies barrier functionality	2 years	evaluation of barrier functionality at the baseline and in enteroids-derived monolayers challenged with pathogens, dead bacteria (as postbiotics), LPS, pharmacological agents, enteral nutrients and to evaluate innate immune response and barrier functionality as previously investigated in fetal enteroids and the contribution of myofibroblast, immune and ENS to the immune response
compare the intestinal barrier functionality in pathological conditions	2 years	comparison of the intestinal barrier functionality in infants complicated by condition of prenatal hypoxia versus non hypoxic infants
Validation of the enteroid NEC model	2 years	Validate the NEC enteroids as an in vitro model for the identification of treatments and prevention of NEC.
functional studies on cellular death	2 years	studies to identify the activation of processes leading to intestinal epithelium necroptosis and/or apoptosis in bacteria challenged and hypoxic conditions
Comparative studies of gene expression	2 years	assess the gene expression profile of tissue, epithelial enteroids and underlying lamina propria derived from NEC, non-NEC (further classified as hypoxic and non-hypoxic infants).; NB: the unit of analysis will be the organoids derived from patients' tissues Investigators expect to be able to derive 1 cell line per patient, and the number of derived organoids will depend from the viability of individual cell lines.