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

NCT ID: NCT05514470

Last Updated: 2025-09-25

Basic Information

• Recruitment Status: WITHDRAWN
• Clinical Phase: NA
• Study Classification: INTERVENTIONAL
• Study Start Date: 2024-11-30
• Study Completion Date: 2027-11-30

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.

Detailed Description

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.

The parameters below will be studied in vitro in cell culture from skin biopsies of patients and control cells:

• Determination of total protein content
• The incorporation of d-methionine, leucine, tyrosine or phenylalanine into proteins
• The study of polysomes profiling
• The study of the assembly of the ribosomal 43S pre-initiation complex
• The phosphorylation of eIF2α and 4EBP and the expression of ATF4
• Ribosome profiling
• Transfer RNA (tRNA) sequencing
• The production of reactive oxygen species (ROS)

The results of these studies will be compared:

• Between patient cells and control cells
• Between genetically corrected patient cells, by stable transfection of the wild-type cDNA of the concerned genes and uncorrected cells
• Between patient cells cultured in medium enriched with the corresponding amino acid.

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

Study Design

• Allocation Method: NA
• Intervention Model: SINGLE_GROUP
• Primary Study Purpose: BASIC_SCIENCE
• Blinding Strategy: NONE

Study Groups

Patients

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

Group Type: EXPERIMENTAL

Skin biopsy

Intervention Type: 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.

Interventions

Skin biopsy

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.

Intervention Type: OTHER

Eligibility Criteria

Inclusion Criteria

• 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

Exclusion Criteria

• 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

• Eligible Sex: ALL
• Accepts Healthy Volunteers: No

Sponsors

URC-CIC Paris Descartes Necker Cochin

OTHER

Sponsor Role: collaborator

Assistance Publique - Hôpitaux de Paris

OTHER

Sponsor Role: lead

Responsible Party

Responsibility Role: SPONSOR

Principal Investigators

Alice HADCHOUEL, MD, PhD

Role: PRINCIPAL_INVESTIGATOR

Assistance Publique - Hôpitaux de Paris

Isabelle SERMET-GAUDELUS, MD, PhD

Role: STUDY_DIRECTOR

Assistance Publique - Hôpitaux de Paris

Locations

Hôpital Necker-Enfants Malades

Paris, , France

Countries

France

Other Identifiers

2024-A01311-46

Identifier Type: OTHER

Identifier Source: secondary_id

APHP220276

Identifier Type: -

Identifier Source: org_study_id