Impact of Mutations in Aminoacyl tRNA Synthetases on Protein Translation and Cellular Stress

NCT05514470 | Withdrawn

• 2 other identifiers: APHP2202762024-A01311-46
• Study Type: interventional
• Target: N/A
• Locations: 1 country
• Sites: 1

Brief Summary

Mutations in the genes encoding cytosolic aminoacyl-tRNA synthetases are responsible for early-onset multisystemic diseases including to varying degrees interstitial lung disease, liver damage, neurological and digestive disorders, and systemic inflammation. These are rare and severe diseases whose pathophysiology is poorly understood. The investigative team hypothesizes that mutations within these genes are responsible for a decrease in protein translation and lead to a cellular stress response similar to that induced by amino acid deprivation. The investigative team also hypothesizes that these alterations could be corrected by high-dose supplementation in the culture medium of the corresponding amino acid. The main objective of the study is to precisely determine the consequences of cytosolic aminoacyl-tRNA synthetase mutations at the cell level on protein translation.

Conditions

Interstitial Lung and Liver Disease; Infantile Liver Failure Syndrome 1; Neurologic, Endocrine and Pancreatic Disease, Multisystem, Infantile-Onset 2; Rajab Interstitial Lung Disease With Brain Calcifications 2

Keywords

Cytosolic aminoacyl-tRNA synthetase; Mutations in the MARS1 gene; Mutations in the LARS1 gene; Mutations in the YARS1 gene; Mutations in the FARSA gene

Outcome Measures

Primary Outcomes (8)

• Determination of total protein content

  Determination of total protein content by Bicinchoninic acid assay.

  Day 0

• Incorporation of d-methionine and d-phenylalanine into proteins

  Incorporation of methionine and phenylalanine by labelled amino-acid fluorescent assays using ready-to-use kits.

  Day 0

• Study of polysomes profiling

  Study of polysome profils by differential sedimentation on sucrose gradients.

  Day 0

• Study of the assembly of the ribosomal 43S pre-initiation complex

  Study of the assembly of the ribosomal 43S pre-initiation complex by co-immunoprecipitation experiments.

  Day 0

• Phosphorylation of eIF2α and 4EBP and the expression of ATF4

  Phosphorylation of eIF2α and 4EBP and the expression of ATF4 by western blot.

  Day 0

• Ribosome profiling

  Ribosome profiling by high throughput sequencing.

  Day 0

• Transfer RNA (tRNA) sequencing

  Transfer RNA (tRNA) sequencing by high throughput sequencing.

  Day 0

• Production of reactive oxygen species (ROS)

  Production of reactive oxygen species (ROS) by fluorescent measurement after cells' incubation with 2',7'- dichlorodihydrofluorescein diacetate (H2DCFDA).

  Day 0

Study Arms (1)

Patients

EXPERIMENTAL

Patients with mutations in genes encoding cytosolic aminoacyl-tRNA synthetases and cared at Necker Hospital.

Other: Skin biopsy

Interventions

Skin biopsy OTHER

A skin biopsy performed on the forearm or thigh depending on the patient's age and wishes, with a biopsy punch with a diameter of 3 to 4 mm depending on the child's age (3 for children under 3 years, 4 beyond). Culture of fibroblasts and immortalization.

Patients

Eligibility Criteria

• Sex: all
• Healthy Volunteers: No
• Age Groups: Child (0-17), Adult (18-64), Older Adult (65+)

You may qualify if:

• Patients carrying mutations in genes encoding cytosolic aminoacyl-tRNA synthetases responsible for a multi-systemic phenotype
• Information and consent of the patient if an adult and of the holders of parental authority if a minor patient and of the minor patient

You may not qualify if:

• \- Non-consent of one of the holders of parental authority or of the minor patient or of adult patient
• Contrôl patients :
• Fibroblasts from control patients without mutation in genes encoding cytosolic aminoacyl-tRNA synthetases, from an existing biological collection. The control patients will be selected according to the age at which the skin biopsy was performed in order to have an age match between the patients and the controls.
• Information and consent of the patient if an adult and of the holders of parental authority if a minor patient and of the minor patient

Sponsors & Collaborators

• Assistance Publique - Hôpitaux de Paris: lead
• URC-CIC Paris Descartes Necker Cochin: collaborator

Study Sites (1)

Hôpital Necker-Enfants Malades

Paris, 75015, France

Location

MeSH Terms

Conditions

Liver Diseases; Neurologic Manifestations; Pancreatic Diseases

Condition Hierarchy (Ancestors)

Digestive System Diseases; Nervous System Diseases; Signs and Symptoms; Pathological Conditions, Signs and Symptoms

Study Officials

• Alice HADCHOUEL, MD, PhD

  Assistance Publique - Hôpitaux de Paris

  PRINCIPAL INVESTIGATOR

• Isabelle SERMET-GAUDELUS, MD, PhD

  Assistance Publique - Hôpitaux de Paris

  STUDY DIRECTOR

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: NA
• Masking: NONE
• Purpose: BASIC SCIENCE
• Intervention Model: SINGLE GROUP
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: June 17, 2022
• First Posted: August 24, 2022
• Study Start: November 1, 2024
• Primary Completion (Estimated): November 1, 2027
• Study Completion (Estimated): November 1, 2027
• Last Updated: September 25, 2025

Record last verified: 2025-09

Data Sharing

• IPD Sharing: Will not share

Locations

FR (1)