mtDNA Mutation Load Analysis in Mesoangioblasts

Recruiting

• Conditions: Mitochondrial Myopathies
• Interventions: Other: in vitro analysis

• Registration Number: NCT05199740
• Lead Sponsor: Maastricht University

Brief Summary

Mitochondrial diseases caused by defects in oxidative phosphorylation (OXPHOS) due to heteroplasmic mitochondrial DNA (mtDNA) mutations are rare (frequency 1/5,000), but severe multi-system disorders. Clinical manifestations are highly variable, but predominantly affect energy demanding tissues, like brain and muscle. Myopathy is a common feature of mtDNA disorders, being present in more than 50% of the mtDNA mutation carriers, and seriously affects patients' general well-being and quality of life. Currently, no treatment is available for these patients, although the induction of muscle regeneration by exercise treatment has been shown to alleviate their myopathy. This implies that these patients can produce muscle fibres that perform better, most likely because the mutation load is lower. Mesoangioblasts (MABs) are myogenic precursors that have been recognized as a source for development of a systemic myogenic stem-cell therapy. Autologous MABs may be feasible for half of the mtDNA mutation carriers of 6 different mtDNA mutations, as their mtDNA mutation load in mesoangioblasts was (nearly) absent (\<10%). However, there are many more mtDNA mutations in the 16.5kb mtDNA and the aim of this study is to determine the mtDNA mutation load in mesoangioblasts of other mtDNA mutation carriers and identify the patients or mutations for which this is a feasible approach.

Detailed Description

Rationale: Mitochondrial diseases caused by defects in oxidative phosphorylation (OXPHOS) due to heteroplasmic mitochondrial DNA (mtDNA) mutations are rare (frequency 1/5,000), but severe multi-system disorders. Clinical manifestations are highly variable, but predominantly affect energy demanding tissues, like brain and muscle. Myopathy is a common feature of mtDNA disorders, being present in more than 50% of the mtDNA mutation carriers, and seriously affects patients' general well-being and quality of life. Currently, no treatment is available for these patients, although the induction of muscle regeneration by exercise treatment has been shown to alleviate their myopathy. This implies that these patients can produce muscle fibres that perform better, most likely because the mutation load is lower. Mesoangioblasts (MABs) are myogenic precursors that have been recognized as a source for development of a systemic myogenic stem-cell therapy, and allogeneic transplantation has been successfully applied to mice and dogs with Duchenne muscular dystrophy. A subsequent phase I/II clinical study in boys with DMD demonstrated that donor MABs treatment was relatively safe, but did not result in clinical improvement, which can partly be attributed to the required use of immunosuppressive agents. The use of autologous MABs would circumvent this and a previous study of our group demonstrated that this is feasible for half of the mtDNA mutation carriers of 6 different mtDNA mutations, as their mtDNA mutation load in mesoangioblasts was (nearly) absent (\<10%). However, there are many more mtDNA mutations in the 16.5kb mtDNA and the aim of this study is to determine the mtDNA mutation load in mesoangioblasts of other mtDNA mutation carriers and identify the patients or mutations for which this is a feasible approach.

Objective: The primary objectives of this project is to assess the mtDNA mutation load in mesoangioblasts of mtDNA mutation carriers and identify which patients display no/low (\<10%) mtDNA mutation load in mesoangioblasts. Secondary objectives aim at determining the proliferation, myogenic differentiation and OXPHOS capacity of mesoangioblasts, their systemic inflammation status and assessment of the mtDNA mutation load in satellite cells.

Study design: Mono-centre observation study. Study population: 30 adult carriers of a heteroplasmic mtDNA point-mutation or large-scale mtDNA deletion (\>2kb).

Intervention: From each participant, a 30mg skeletal muscle biopsy and a 20ml venous blood sample will be collected.

Main study parameters/endpoints: Assess the mtDNA mutation load in skeletal muscle derived mesoangioblasts.

Study Timeline

• Study First Submitted: 2021-12-17
• Study First Submitted QC: 2022-01-06
• Study First Posted: 2022-01-20
• Study Start: 2022-12-20
• Status Verified: 2024-01
• Last Update Submitted: 2024-01-22
• Last Update Posted: 2024-01-24
• Primary Completion: 2025-07-01
• Study Completion: 2026-07-01

Recruitment & Eligibility

• Status: RECRUITING
• Sex: All
• Target Recruitment: 30

Inclusion Criteria

Not provided

Exclusion Criteria

Not provided

Study & Design

• Study Type: OBSERVATIONAL
• Study Design: Not specified

Arm && Interventions

Group	Intervention	Description
mtDNA mutation carriers	in vitro analysis	Carriers of a pathogenic mtDNA mutation

Primary Outcome Measures

Name	Time	Method
Assess mtDNA mutation load in mesoangioblasts	1 day	Isolate mesoangioblasts and quantify mtDNA mutation load using GeneScan analysis

Secondary Outcome Measures

Name	Time	Method
Assess level of systemic inflammation marker SDF-1 in blood plasma	1 day	SDF-1 measurement in plasma (pg/ml)
Assess level of systemic inflammation marker TNFa in blood plasma	1 day	TNFa measurement in plasma (pg/ml)
Assess mitochondrial function in mesoangioblasts	1 day	Assess mtDNA copynumber and OXPHOS capacity in mesoangioblasts
Assess level of systemic inflammation marker IL-6 in blood plasma	1 day	IL-6 measurement in plasma (pg/ml)
Assess myogenic differentiation capacity of mesoangioblasts	1 day	Assess myogenic fusion index of mesoangioblasts isolated from a muscle biopsy
Assess mtDNA mutation load in satellite cells	1 day	Isolate satellite cells and quantify mtDNA mutation load using GeneScan analysis
Assess level of systemic tissue damage marker CK in blood plasma	1 day	CK measurement in plasma (U/l)

Trial Locations

Locations (1)

Maastricht University; 🇳🇱 Maastricht, Netherlands