Endometriosis
Conditions
Keywords
Diaphragmatic endometriosis, Surgical nodules resection
Brief summary
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.
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.
Interventions
OTHERDuring 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.
Sponsors
Assistance Publique - Hôpitaux de Paris
URC-CIC Paris Descartes Necker Cochin
UMR INSERM 1124
Study design
Observational model
COHORT
Time perspective
PROSPECTIVE
Eligibility
Sex/Gender
FEMALE
Age
18 Years to No maximum
Healthy volunteers
No
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
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Molecular characteristics | Inclusion | 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). |
| Cellular characteristics | Inclusion | 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). |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Presence of diaphragm-derived myoblasts | Inclusion | 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. |
Countries
France
Contacts
CONTACTChristelle NGUYEN, MD, PHD
CONTACTMarie BENHAMMANI-GODARD
STUDY_DIRECTORChristelle NGUYEN, MD, PHD
Physical Medicine and Rehabilitation Department - Cochin hospital
PRINCIPAL_INVESTIGATORMarco ALIFANO, MD, PHD
Department of Thoracic Surgery - Cochin Hospital
Outcome results
None listed