Role of ACTG2 Variants in Smooth Muscle Determination and Function in Pediatric Intestinal Pseudo-obstruction.

N/ANot Yet RecruitingNCT06687564

University Hospital, Grenoble4 enrolled

Overview

The primary objective of this study is to describe the transcriptional impact of R178, R257, R40 or A136 variants of the ACTG2 gene on iPS differentiation mechanisms up to organoids derived from PIPO patient samples versus those derived from control / WT patients (generation of IPS from cultured cell lines), at different stages of their experimental ex vivo development.

Recruited patients will be sampled during a consultation: a blood sample and a biopsy will be taken directly from the patient. Once these samples have been taken, they will be cultured to be reprogrammed into iPS cells, then grown and differentiated into intestinal organoids. Various experiments will be carried out (as described in the outcomes) to identify at molecular, cellular and tissue level the mechanisms altered in patients with an R178, R257, R40 or A136 variant, assessing their consequence(s) on the development and functionality of the digestive mesenchyme.

Study Type

INTERVENTIONAL

Allocation

NON_RANDOMIZED

Purpose

BASIC_SCIENCE

Masking

NONE

Enrollment

Conditions

PIPO

Interventions

BiopsyPROCEDURE

A skin biopsy and a blood sample will be taken to culture the iPS cells and intestinal organoids.

Eligibility

Sex: ALLMin age: 4 Years

Medical Language ↔ Plain English

Inclusion Criteria: PIPO Population: 1. Minor or adult patient ≥ 4 years of age 2. Patient with PIPO before age 18 3. Male or female 4. Patient with PIPO meeting at least 2 of the ESPGHAN criteria (Thapar et al 2018) and carrying the R178, R257, R40 or A136 mutation of the ACTG2 gene. 5. Patient whose assent has been obtained and whose legal guardians have given their written informed consent 6. Patient affiliated to the French Social Security system or benefiting from an equivalent plan WT population: \- iPS cell lines MS573 or WT8288 or 202CT or SD378M, from the Nantes University Hospital biological collection and generated from samples from control patients without POIC who have consented to donate their samples. Exclusion Criteria: PIPO population : 1. Patients with a history of radiotherapy treatment 2. Patient with lymphocyte lineage damage

Locations (3)

Phymedexp Inserm U1046 - Cnrs Umr 9214

Montpellier, France

Tens - Inserm Un Umr 1235

Nantes, France

AP-HP Hôpital Robert Debré

Paris, France

Outcomes

Primary Outcomes

Description of the transcriptional impact of R178, R257, R40 or A136 variants of the ACTG2 gene

Difference in transcript expression (%) in RNASeq transcriptomic analysis between samples carrying the R178, R257, R40 or A136 variants of the ACTG2 gene on the differentiation mechanisms of iPS up to organoids derived from PIPO patient samples versus those derived from control patients

Time frame: At different stages of their experimental ex vivo development (mesenchymal progenitors, determined smooth muscle cells, differentiated smooth muscle cells and 3D organization of smooth muscle) through study completion, an average of 5 years

Secondary Outcomes

Evaluate the impact of R178, R257, R40 or A136 variants of the ACTG2 gene on gastrointestinal contractile function between mutant versus WT organoids.

Difference (%) in isometric contraction force between mutant versus WT organoids by electrical stimulation and specific agonist.

Evaluate the immunofluorescence labeling differential between mutant and WT cells from IPS and organoids

Difference in fluorescence (%) between mutant and WT cells as cell stages from iPS to organoids

Evaluate the impact of R178, R257, R40 or A136 mutations of the ACTG2 gene on the actin network of fibroblasts derived from skin samples of diseased patients versus those of WTpatients.

Difference in actin network labeling by immunofluorescence (%) between skin fibroblasts from PIPO and WT patients.

Central Contacts

John Rendu, Dr

CONTACT

0476765573 jrendu@chu-grenoble.fr

Mandy Leger

CONTACT

0476768410 mleger@chu-grenoble.fr

Data from ClinicalTrials.gov

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