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
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.
Study Type
INTERVENTIONAL
Allocation
NON_RANDOMIZED
Purpose
OTHER
Masking
NONE
Enrollment
60
* 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
Olivier FOUQUET
Angers, France
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
Time frame: 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
Time frame: 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
Time frame: 1 month
Level of tissue expression of the genes and coding for the proteins Fis1
Level expression of Fis1 by RT-qPCR
Time frame: 1 month
Level of tissue expression of the genes and coding for the proteins Drp1
Level expression of Drp1 by RT-qPCR
Time frame: 1 month
Level of tissue expression of the genes and coding for the proteins Nfr1
Level expression of Nfr1 by RT-qPCR
Time frame: 1 month
Level of tissue expression of the genes and coding for the proteins Tfam
Level expression of Tfam by RT-qPCR
Time frame: 1 month
Level of tissue expression of the genes and coding for the proteins PGC1⍺
Level expression of PGC1⍺ by RT-qPCR
Time frame: 1 month
Analysis of mitochondrial network
This platform is for informational purposes only and does not constitute medical advice. Always consult a qualified healthcare professional.
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.
Time frame: 1 month
Protein of remodelling and constitution of extracellular matrix: Metalloprotease MMp2
Level expression of Metalloprotease MMp2 by western blot
Time frame: 1 month
Proteins of remodelling and constitution of extracellular matrix: Timp 1/2
Level expression of Timp 1/2 by western blot
Time frame: 1 month
Proteins of remodelling and constitution of extracellular matrix: Collagene I/III
Level expression of Collagene I/III by western blot
Time frame: 1 month.
Protein of remodelling and constitution of extracellular matrix: Elastine
Level expression of Elastine by western blot
Time frame: 1 month
Proteins of survival cell: Bcl2/Bax
Level expression of Bcl2/Bax by western blot
Time frame: 1 month
Proteins of survival cell: Cytochrome C
Level expression of Cytochrome C by western blot
Time frame: 1 month
Proteins of oxydative stress: NADPH
Level expression of NADPH by western blot
Time frame: 1 month
Proteins of oxydative stress: OxyD
Level expression of OxyD by western blot
Time frame: 1 month
Proteins of oxydative stress: Sod 1/2
Level expression of Sod 1/2 by western blot
Time frame: 1 month
Proteins of Smooth Muscle Cell reactivity: Myh11
Level expression of Myh11 by western blot
Time frame: 1 month
Proteins of Smooth Muscle Cell reactivity: Acta 2
Level expression of Acta 2 by western blot
Time frame: 1 month
Proteins of Smooth Muscle Cell reactivity: MLC20
Level expression of MLC20 by western blot
Time frame: 1 month
Proteins of Smooth Muscle Cell reactivity: Rock1
Level expression of Rock1 by western blot
Time frame: 1 month
Proteins of Smooth Muscle Cell reactivity: Rhoa
Level expression of Rhoa by western blot
Time frame: 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.
Time frame: 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.
Time frame: 2 years