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Mitochondrial Dysfunctions Driving Insulin Resistance

Recruiting
Conditions
Mitochondrial Diseases
Mitochondrial Myopathies
Mitochondrial Disorders
Registration Number
NCT06080581
Lead Sponsor
Rigshospitalet, Denmark
Brief Summary

The overarching aim of this observational study is to characterize muscle mitochondrial defects in individuals harboring pathogenic mitochondrial DNA (mtDNA) mutations associated with an insulin-resistant phenotype.

In a case-control design, individuals with pathogenic mtDNA mutations will be compared to controls matched for sex, age, and physical activity level. Participants will attend a screening visit and two experimental trials including:

* An oral glucose tolerance test

* A hyperinsulinemic-euglycemic clamp combined with measurements of femoral artery blood flow and arteriovenous difference of glucose

* Muscle biopsy samples

Detailed Description

Background: Peripheral insulin resistance is a major risk factor for metabolic diseases such as type 2 diabetes. Skeletal muscle accounts for the majority of insulin-stimulated glucose disposal, hence restoring insulin action in skeletal muscle is key in the prevention of type 2 diabetes. Mitochondrial dysfunction is implicated in the etiology of muscle insulin resistance. Also, as mitochondrial function is determined by its proteome quantity and quality, alterations in the muscle mitochondrial proteome may play a critical role in the pathophysiology of insulin resistance. However, insulin resistance is multifactorial in nature and whether mitochondrial derangements are a cause or a consequence of impaired insulin action is unclear. In recent years, the study of humans with genetic mutations has shown enormous potential to establish the mechanistic link between two physiological variables; indeed, if the mutation has a functional impact on one of those variables, then the direction of causality can be readily ascribed. Mitochondrial myopathies are genetic disorders of the mitochondrial respiratory chain affecting predominantly skeletal muscle. Mitochondrial myopathies are caused by pathogenic mutations in either nuclear or mitochondrial DNA (mtDNA), which ultimately lead to mitochondrial dysfunction. Although the prevalence of mtDNA mutations is just 1 in 5,000, the study of patients with mtDNA defects has the potential to provide unique information on the pathogenic role of mitochondrial derangements that is disproportionate to the rarity of affected individuals. The m.3243A\>G mutation in the MT-TL1 gene encoding the mitochondrial leucyl-tRNA 1 gene is the most common mutation leading to mitochondrial myopathy in humans. The m.3243A\>G mutation is associated with impaired glucose tolerance and insulin resistance in skeletal muscle. Most importantly, insulin resistance precedes impairments of β-cell function in carriers of the m.3243A\>G mutation, making these patients an ideal human model to study the causative nexus between muscle mitochondrial dysfunction and insulin resistance. Thus, a comprehensive characterization of mitochondrial functional defects and the associated proteome alterations in patients harboring a mtDNA mutation associated with an insulin-resistant phenotype may elucidate the causal nexus between mitochondrial derangements and insulin resistance. Also, as mitochondrial dysfunction exhibits many faces (e.g. reduced oxygen consumption rate, impaired ATP synthesis, overproduction of reactive oxygen species, altered membrane potential), such an approach may clarify which features of mitochondrial dysfunction play a prominent role in the pathogenesis of insulin resistance.

Objective: To characterize muscle mitochondrial defects in individuals harboring pathogenic mitochondrial DNA (mtDNA) mutations associated with an insulin-resistant phenotype.

Study design: Case-control study in individuals with pathogenic mtDNA mutations (n=15) and healthy controls (n=15) matched for sex, age, and physical activity level.

Endpoint: Differences between individuals with pathogenic mtDNA mutations and controls.

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
Primary Outcome Measures
NameTimeMethod
Muscle mitochondrial respirationBaseline

Mitochondrial O2 flux is measured by high-resolution respirometry in permeabilized fibers from muscle biopsy samples

Muscle mitochondrial reactive oxygen species (ROS) productionBaseline

Mitochondrial H2O2 emission rates are measured by high-resolution fluorometry in permeabilized fibers from muscle biopsy samples

Skeletal muscle insulin sensitivity90-150 minutes after initiation of the hyperinsulinemic euglycemic clamp

Insulin-stimulated muscle glucose uptake is determined by the hyperinsulinemic-euglycemic clamp method integrated with measurements of femoral artery blood flow and arteriovenous difference of glucose

Muscle mitochondrial proteomeBaseline

Mitochondrial proteome signatures are determined by mass spectrometry-based proteomics in muscle biopsy samples

Whole-body insulin sensitivity90-150 minutes after initiation of the hyperinsulinemic euglycemic clamp

Whole-body insulin sensitivity is determined by the hyperinsulinemic-euglycemic clamp method

Secondary Outcome Measures
NameTimeMethod
Muscle mtDNA heteroplasmyBaseline

mtDNA mutation load is measured in muscle biopsy samples from the patients with mitochondrial myopathy

Muscle insulin signalingBefore (baseline) and 0-150 minutes after initiation of a hyperinsulinemic-euglycemic clamp

Insulin-mediated changes in the abundance of (phosphorylated) proteins modulating insulin action are measured by immunoblotting in muscle and fat biopsy samples

Muscle integrated stress response signaling proteinsBaseline

Abundance of (phosphorylated) proteins modulating the integrated stress response pathway is measured by immunoblotting in muscle biopsy samples.

Glucose tolerance0-180 minutes after ingestion of an oral glucose solution

Glucose tolerance is determined by the plasma glucose response curve measured during an oral glucose tolerance test

Muscle release of FGF21 and GDF15Before (baseline) and 0-150 minutes after initiation of a hyperinsulinemic-euglycemic clamp

Skeletal muscle production of FGF21 and GDF15 is determined by measurements of femoral artery blood flow and arteriovenous difference of plasma FGF21 and GDF15

Beta cell function0-180 minutes after ingestion of an oral glucose solution

Beta cell function is determined by measurements of plasma insulin and insulin C-peptide during an oral glucose tolerance test

Muscle integrated stress response genesBaseline

mRNA content of genes governing the integrated stress response pathway is measured by Real-Time PCR in muscle biopsy samples.

Trial Locations

Locations (1)

Rigshospitalet

🇩🇰

Copenhagen, Denmark

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