MITAORTA - Role of Mitochondrial Dynamic in Aneurysm and Dissection of Ascending Thoracic Aorta

Study Overview
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Brief Summary
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The main objective is to compare the mitochondrial dynamic between patients operated for aneurysm of ascending aorta or type A aortic dissection (AAD) or control group

Detailed Description
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In an aortic aneurysm process, the alteration of the extracellular matrix (ECM) as well as the apoptosis of the smooth muscle cells are due to inflammatory phenomena and oxydative stress, involving mitochondria which has a key place within cells. Mitochondrial fusion and fission constitute mitochondrial dynamic and are involved in the mechanisms described above. The alteration of mitochondrial dynamics has been demonstrated in many pathologies, in particular neurological, cancer and cardiovascular disease and generally occurs in favor of fission. In a mouse model (FASEB J, 2021, Robert P ), the role of mitochondrial fusion has been demostrated as a protective factor against hypertension in resistance arteries and a deletion of OPA1 (optic Atrophy 1) fusion protein may lead to aneurysm until aortic dissection. The results of this experimental study suggest a role of the alteration of mitochondrial dynamic in the development of aneurysm and aortic dissection.

Official Title
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MITAORTA - Role of Mitochondrial Dynamic in Aneurysm and Dissection of Ascending Thoracic Aorta

Conditions
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Aortic Aneurysm and Dissection

Intervention / Treatment
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* Other: Mitochondrial dynamic analysis in the aorta samples and metabolomic profiling in the aortic diseases


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Aneurysm aortic group: patients treated for an aneurysm of the ascending thoracic aorta with surgical indication according to the ESC guidelines (2014). Aortic dissection group: patients treated for type A acute aortic dissection or intramural hematoma of the ascending thoracic aorta in emergency. Control group: patients operated for aortic valve replacement (little aortic sample before closing aortotomy) or coronary artery bypass grafting which the use of a saphenous graft and the performance of a proximal anastomosis on the ascending aorta is planned Exclusion Criteria: Patients under 18 years old Other acute aortic syndromes (penetrating ulcers, iatrogenic or traumatic dissections) Patients treated for aortic valve replacement in the context of infective endocarditis Patients treated for emergency aortic valve replacement or coronary bypass surgery** Pregnant, parturient and breastfeeding women Patients protected by an administrative or judicial measure (curatorship, guardianship) Patients receiving psychiatric care under duress Adults subject to a legal protection measure. Patients whose the samples planned for the study could not be taken; Patients in the control group whose tissue sampling will not be performed.

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 90 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Other
Allocation: Non-Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: Aneurysm and Dissection of Ascending Thoracic Aorta
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: Aneurysm group<br>Patients operated for aneurysm of the ascending aorta according the guidelines on the diagnosis and treatment of aortic diseases (European Society of Cardiology - 2014). | Other: Mitochondrial dynamic analysis in the aorta samples and metabolomic profiling in the aortic diseases<br>* For each patient: a segment of aorta will be sampled and cutted into 4 parts 1 fragment placed in a Falcon tube containing DMEM (Dulbecco's Modified Eagle Medium, Thermo Fisher®), temporarily stored at + 4°C, for primary culture of smooth muscle cells will allow analysis of the mitochondrial network. 1 fragment placed in a Falcon tube containing Allprotect Tissue Reagent (QIAGEN®), stored at -80°C for gene (RT-PCR) and protein (Western Blot) analysis. 2 fragments each placed in dry cryotube stored at - 80°C will be used for metabolomic analysis. For each patient, 2 arterial blood samples will be collected before general anaesthesia One tube of whole blood stored at -80°C for metabolomic analysis. One tube of blood stored at -80°C for plasma cytokines<br>|
| Active Comparator: Type A aortic dissection group<br>Patients operated for type A aortic dissection according the guidelines on the diagnosis and treatment of aortic diseases (European Society of Cardiology - 2014). | Other: Mitochondrial dynamic analysis in the aorta samples and metabolomic profiling in the aortic diseases<br>* For each patient: a segment of aorta will be sampled and cutted into 4 parts 1 fragment placed in a Falcon tube containing DMEM (Dulbecco's Modified Eagle Medium, Thermo Fisher®), temporarily stored at + 4°C, for primary culture of smooth muscle cells will allow analysis of the mitochondrial network. 1 fragment placed in a Falcon tube containing Allprotect Tissue Reagent (QIAGEN®), stored at -80°C for gene (RT-PCR) and protein (Western Blot) analysis. 2 fragments each placed in dry cryotube stored at - 80°C will be used for metabolomic analysis. For each patient, 2 arterial blood samples will be collected before general anaesthesia One tube of whole blood stored at -80°C for metabolomic analysis. One tube of blood stored at -80°C for plasma cytokines<br>|
| Sham Comparator: Control group<br>Patients without aortic aneurysm or aortic dissection operated for aortic valve replacement (AVR) and/or coronary artery bypass with a saphenous vein graft for proximal aortic anastomosis to collect the aortic sample. For patients operated for AVR, an aortic sample will be collected before closing the aorta. | Other: Mitochondrial dynamic analysis in the aorta samples and metabolomic profiling in the aortic diseases<br>* For each patient: a segment of aorta will be sampled and cutted into 4 parts 1 fragment placed in a Falcon tube containing DMEM (Dulbecco's Modified Eagle Medium, Thermo Fisher®), temporarily stored at + 4°C, for primary culture of smooth muscle cells will allow analysis of the mitochondrial network. 1 fragment placed in a Falcon tube containing Allprotect Tissue Reagent (QIAGEN®), stored at -80°C for gene (RT-PCR) and protein (Western Blot) analysis. 2 fragments each placed in dry cryotube stored at - 80°C will be used for metabolomic analysis. For each patient, 2 arterial blood samples will be collected before general anaesthesia One tube of whole blood stored at -80°C for metabolomic analysis. One tube of blood stored at -80°C for plasma cytokines<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Level of tissue expression of the genes and coding for the proteins (Optic Atrophy 1) OPA1 | Level expression of, (Optic Atrophy 1) OPA1 by RT-qPCR | 1 month | 
| Level of tissue expression of the genes and coding for the proteins MFN1 (Mitofusin 1) | Level expression of MFN1 (Mitofusin 1) by RT-qPCR | 1 month | 
| Level of tissue expression of the genes and coding for the proteins MFN2 (Mitofusin 2) | Level expression of MFN2 (Mitofusin 2) by RT-qPCR | 1 month | 
| Level of tissue expression of the genes and coding for the proteins Fis1 | Level expression of Fis1 by RT-qPCR | 1 month | 
| Level of tissue expression of the genes and coding for the proteins Drp1 | Level expression of Drp1 by RT-qPCR | 1 month | 
| Level of tissue expression of the genes and coding for the proteins Nfr1 | Level expression of Nfr1 by RT-qPCR | 1 month | 
| Level of tissue expression of the genes and coding for the proteins Tfam | Level expression of Tfam by RT-qPCR | 1 month | 
| Level of tissue expression of the genes and coding for the proteins PGC1⍺ | Level expression of PGC1⍺ by RT-qPCR | 1 month | 
| Analysis of mitochondrial network | To analyze the mitochondrial network, vascular smooth muscle cells will be extracted from the wall of aorta samples and seeded in Petri dish. When 80% confluence is obtained, cells will be incubated with a green fluorescent marker (Mitotacker Green Probes) and 3D fluorescence microscopy will be used. Analysis of mitochondrial network will be done after characterization of mitochondrial shapes and distribution in the different aorta samples. | 1 month | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Protein of remodelling and constitution of extracellular matrix: Metalloprotease MMp2 | Level expression of Metalloprotease MMp2 by western blot | 1 month | 
| Proteins of remodelling and constitution of extracellular matrix: Timp 1/2 | Level expression of Timp 1/2 by western blot | 1 month | 
| Proteins of remodelling and constitution of extracellular matrix: Collagene I/III | Level expression of Collagene I/III by western blot | 1 month. | 
| Protein of remodelling and constitution of extracellular matrix: Elastine | Level expression of Elastine by western blot | 1 month | 
| Proteins of survival cell: Bcl2/Bax | Level expression of Bcl2/Bax by western blot | 1 month | 
| Proteins of survival cell: Cytochrome C | Level expression of Cytochrome C by western blot | 1 month | 
| Proteins of oxydative stress: NADPH | Level expression of NADPH by western blot | 1 month | 
| Proteins of oxydative stress: OxyD | Level expression of OxyD by western blot | 1 month | 
| Proteins of oxydative stress: Sod 1/2 | Level expression of Sod 1/2 by western blot | 1 month | 
| Proteins of Smooth Muscle Cell reactivity: Myh11 | Level expression of Myh11 by western blot | 1 month | 
| Proteins of Smooth Muscle Cell reactivity: Acta 2 | Level expression of Acta 2 by western blot | 1 month | 
| Proteins of Smooth Muscle Cell reactivity: MLC20 | Level expression of MLC20 by western blot | 1 month | 
| Proteins of Smooth Muscle Cell reactivity: Rock1 | Level expression of Rock1 by western blot | 1 month | 
| Proteins of Smooth Muscle Cell reactivity: Rhoa | Level expression of Rhoa by western blot | 1 month | 
| Analysis of Plasma Metabolomes | 154 metabolites will be measured in plasma samples. Plasma data blocks will be submitted to multiblock orthogonal component analysis (MOCA) after normalizing concentrations of plasma samples by their weights. | 2 years | 
| Analysis of Aorta Metabolomes | 136 metabolites will be measured in plasma samples. Plasma data blocks will be submitted to multiblock orthogonal component analysis (MOCA) after normalizing concentrations of aorta samples by their weights. | 2 years | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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extracellular matrix, cell survival, oxydative stress, mitochondrial fusion, metabolomic profiling of aortic aneurym, metabolomic profilng of aortic dissection