" Development of Primary Cultures of Diaphragmatic Myoblasts for Basic Research Purposes " (MYODiaph)

Neuromuscular diseases (NMDs) affecting motor neurons (MN) induce progressive muscle denervation, and become fatal when respiratory muscles e.g. the diaphragm are affected and can no longer contract. In some cases, such as Charcot's disease (Amyotrophic lateral sclerosis-ALS), there is no cure and patients die due to respiratory failure few years after diagnosis. Investigations on NMD-induced alterations of respiratory muscles in humans are limited notably by the absence of available in vitro model based on cell cultures of diaphragm-derived myoblasts. Yet, this cell tool is likely to help in developing original therapies to limit diaphragm muscle atrophy and dysfunction in NMD. To date, only cell cultures of human myoblasts obtained from limb muscles are available, making difficult to transpose results to the diaphragm.

Thus, in the present project, we propose to :

1. originally develop primary cultures of myoblasts from human diaphragm, obtained from surgical resection of diaphragmatic endometriosis,
2. characterize them in terms of differentiation status (Histology, IF), metabolism (Metabolomics by NMR, cell respiration), and gene expression (RNASeq), in comparison to primary myoblasts cultures derived from the deltoid already available in the team.

This project will provide an original new tool and important data on the specificity of diaphragm-derived myoblasts, compared to limb muscle-derived myoblasts with the long-term perspective of opening new therapeutical pathways for patients with severe NMDs.

Study Overview

• Status: Not yet recruiting
• Conditions: Endometriosis
• Intervention / Treatment: Other: During the surgery planned as part of the routine care of the patient, a part of the tissue sample will be saved in order to carry out the analysis planned for the research.

Detailed Description

Neuromuscular diseases (NMDs) affecting motor neurons (MN) induce progressive muscle denervation, and become fatal when respiratory muscles e.g. the diaphragm are affected and can no longer contract. In some cases, such as Charcot's disease (Amyotrophic lateral sclerosis-ALS), there is no cure and patients die due to respiratory failure few years after diagnosis. Investigations on NMD-induced alterations of respiratory muscles in humans are limited notably by the absence of available in vitro model based on cell cultures of diaphragm-derived myoblasts. Yet, this cell tool is likely to help in developing original therapies to limit diaphragm muscle atrophy and dysfunction in NMD. To date, only cell cultures of human myoblasts obtained from limb muscles are available, making difficult to transpose results to the diaphragm.

Thus, in the present project, we propose to :

1. originally develop primary cultures of myoblasts from human diaphragm, obtained from surgical resection of diaphragmatic endometriosis,
2. characterize them in terms of differentiation status (Histology, IF), metabolism (Metabolomics by NMR, cell respiration), and gene expression (RNASeq), in comparison to primary myoblasts cultures derived from the deltoid already available in the team.

This project will provide an original new tool and important data on the specificity of diaphragm-derived myoblasts, compared to limb muscle-derived myoblasts with the long-term perspective of opening new therapeutical pathways for patients with severe NMDs.

• Study Type: Observational
• Enrollment (Estimated): 10

Contacts and Locations

• Study Contact: Name: Marie Benhammani-Godard; Phone Number: 0033158411190; Email: marie.godard@aphp.fr
• Study Contact Backup: Name: Christelle NGUYEN, MD, PhD; Phone Number: 00 33 1 58 41 29 45; Email: christelle.nguyen2@aphp.fr

Study Locations

• France
  • IDF
    • Paris, IDF, France, 75014
      • Department of Thoracic Surgery - Cochin Hospital
      • Contact: Christelle NGUYEN, MD, PhD; Phone Number: 00 33 1 58 41 29 45; Email: christelle.nguyen2@aphp.fr
      • Contact: Marco ALIFANO, MD, PhD; Email: marco.alifano@aphp.fr

Participation Criteria

Eligibility Criteria

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

• Sampling Method: Non-Probability Sample

Study Population

Patients with diaphragmatic endometriosis requiring surgical nodules resection in the diaphragm.

Description

Inclusion Criteria:

• Age ≥ 18 years
• diaphragmatic endometriosis requiring surgical resection of the nodules in the diaphragm.
• Collection of non-opposition

Exclusion Criteria

• Inability to speak and/or read French
• Patients under tutor or curatorship
• Protected adults

Study Plan

How is the study designed?

Number of groups / cohorts

1

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Patients; Patients with endometriosis requiring surgical nodules resection in the diaphragm.	Other: During the surgery planned as part of the routine care of the patient, a part of the tissue sample will be saved in order to carry out the analysis planned for the research.; During the surgery planned as part of the routine care of the patient, a part of the tissue sample will be saved in order to carry out the analysis planned for the research.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Molecular characteristics	The primary endpoint will be the identification of molecular and cellular signatures specific of the diaphragm as compared with human deltoid muscle cultures, notably based on the study of : differentiation, by studying the course of myoblast differentiation and fusion in polynucleated myotubes (IF Desmin, Slow and fast Myosin Heavy Chain),; energy metabolism (extra- and intra-cellular metabolomics by NMR, cell respiration by the Seahorse technology),; gene regulation, by analysing the transcriptomic signature (RNAseq).	Inclusion
Cellular characteristics	The primary endpoint will be the identification of molecular and cellular signatures specific of the diaphragm as compared with human deltoid muscle cultures, notably based on the study of : differentiation, by studying the course of myoblast differentiation and fusion in polynucleated myotubes (IF Desmin, Slow and fast Myosin Heavy Chain),; energy metabolism (extra- and intra-cellular metabolomics by NMR, cell respiration by the Seahorse technology),; gene regulation, by analysing the transcriptomic signature (RNAseq).	Inclusion

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Presence of diaphragm-derived myoblasts	The secondary endpoint will be to build a library of molecular and cellular signatures for the major respiratory muscle i.e. the diaphragm in human. Decisive differences between cellular and molecular signatures across muscles (Diaphragm and Deltoid) will be analysed by principal component analysis.	Inclusion

Collaborators and Investigators

• Sponsor: Assistance Publique - Hôpitaux de Paris
• Collaborators: URC-CIC Paris Descartes Necker Cochin; UMR INSERM 1124
• Investigators: Study Director: Christelle NGUYEN, MD, PHD, Physical Medicine and Rehabilitation Department - Cochin hospital; Principal Investigator: Marco ALIFANO, MD, PHD, Department of Thoracic Surgery - Cochin Hospital

Study record dates

Study Major Dates

• Study Start (Estimated): February 1, 2026
• Primary Completion (Estimated): April 1, 2026
• Study Completion (Estimated): May 1, 2026

Study Registration Dates

• First Submitted: January 14, 2026
• First Submitted That Met QC Criteria: January 27, 2026
• First Posted (Actual): February 2, 2026

Study Record Updates

• Last Update Posted (Actual): February 2, 2026
• Last Update Submitted That Met QC Criteria: January 27, 2026
• Last Verified: January 1, 2026

More Information

Terms related to this study

• Keywords: Diaphragmatic endometriosis; Surgical nodules resection
• Additional Relevant MeSH Terms: Urogenital Diseases; Genital Diseases; Female Urogenital Diseases; Female Urogenital Diseases and Pregnancy Complications; Genital Diseases, Female; Endometriosis
• Other Study ID Numbers: APHP250977; 2024-A02556-41 (Other Identifier: Ministry of Health - France)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: UNDECIDED

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No