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

Impact of Loss-of-function Mutations of Genes Encoding Cytosolic Aminoacyl-tRNA Synthetases on Protein Translation and Responses to Cellular Stress

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.

Study Overview

• Status: Withdrawn
• 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
• Intervention / Treatment: Other: Skin biopsy

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.

• Study Type: Interventional
• Phase: Not Applicable

Contacts and Locations

Study Locations

• France
  • Paris, France, 75015
    • Hôpital Necker-Enfants Malades

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Child; Adult; Older Adult
• Accepts Healthy Volunteers: No

Description

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

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: N/A
• Interventional Model: Single Group Assignment
• Masking: None (Open Label)

Number of Arms

1

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Patients; Patients with mutations in genes encoding cytosolic aminoacyl-tRNA synthetases and cared at Necker Hospital.	Other: 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.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
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

Collaborators and Investigators

• Sponsor: Assistance Publique - Hôpitaux de Paris
• Collaborators: URC-CIC Paris Descartes Necker Cochin
• Investigators: Principal Investigator: Alice HADCHOUEL, MD, PhD, Assistance Publique - Hôpitaux de Paris; Study Director: Isabelle SERMET-GAUDELUS, MD, PhD, Assistance Publique - Hôpitaux de Paris

Study record dates

Study Major Dates

• Study Start (Estimated): November 1, 2024
• Primary Completion (Estimated): November 1, 2027
• Study Completion (Estimated): November 1, 2027

Study Registration Dates

• First Submitted: June 17, 2022
• First Submitted That Met QC Criteria: August 22, 2022
• First Posted (Actual): August 24, 2022

Study Record Updates

• Last Update Posted (Estimated): September 25, 2025
• Last Update Submitted That Met QC Criteria: September 22, 2025
• Last Verified: September 1, 2025

More Information

Terms related to this study

• 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
• Additional Relevant MeSH Terms: Nervous System Diseases; Digestive System Diseases; Pathological Conditions, Signs and Symptoms; Signs and Symptoms; Neurologic Manifestations; Liver Diseases; Pancreatic Diseases
• Other Study ID Numbers: APHP220276; 2024-A01311-46 (Other Identifier: ID-RCB number)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No