Protheomic and adipo/cytokine biomarker analysis of unstable carotid atheroma plaque
- Autores: Ajla Alibalic
- Directores de la Tesis: Cristóbal Richart Jurado (dir. tes.), M Teresa Auguet Quintillà (dir. tes.)
- Lectura: En la Universitat Rovira i Virgili ( España ) en 2022
- Idioma: inglés
- Número de páginas: 186
- Tribunal Calificador de la Tesis: Ramon Miralles Basseda (presid.), Josep Maria Alegret Colomé (secret.), Cristina Tural i Llàcher (voc.)
- Materias:
- Enlaces
- Tesis en acceso abierto en: TDX
- Resumen
- español
La arteriosclerosis es un proceso patológico que provoca manifestaciones clínicas de enfermedades vasculares. Los objetivos de esta tesis son identificar posibles biomarcadores de aterosclerosis carotídea (AC) en muestras de secretoma de pacientes con placa carotídea inestable y estudiar el posible papel de algunas adipo/citoquinas en la estabilidad de las placas ateroscleróticas. También analizamos los niveles de varias adipo/citocinas pro y antiinflamatorias en suero y secretoma de placa inestable según la presencia de factores de riesgo cardiovascular. En el primer estudio, realizamos un análisis proteómico cuantitativo en las muestras de secretoma obtenidas de placas ateroscleróticas carotídeas humanas mediante endarterectomía carotídea. En un segundo estudio, analizamos los niveles de varias adipo/citocinas pro y antiinflamatorias en suero y secretoma de placa inestable mediante ensayos inmunoabsorbentes ligados a enzimas. Se detectaron algunos cambios proteómicos en los secretomas de las placas ateroscleróticas carotídeas (CAP) en comparación con las arterias mamarias no ateroscleróticas (AMNE). Las proteínas aumentadas en los secretomas de CAP fueron la defensina 1 de neutrófilos, la apolipoproteína E, la clusterina y la glicoproteína alfa-2 de zinc.
- català
L'arteriosclerosi és un procés patològic que provoca manifestacions clíniques de malalties vasculars. Els objectius daquesta tesi són identificar possibles biomarcadors d?aterosclerosi carotídia (AC) en mostres de secretoma dels pacients amb placa carotídia inestable i estudiar el possible paper d’algunes adipo/citoquines en l’estabilitat de les plaques ateroscleròtiques. També analitzem els nivells de diverses adipes/citocines pro i antiinflamatòries en sèrum i secretoma de placa inestable segons la presència de factors de risc cardiovascular. En el primer estudi, realitzem una anàlisi proteòmica quantitativa a les mostres de secretoma obtingudes de plaques ateroscleròtiques carotídies humanes mitjançant endarterectomia carotídia. En un segon estudi, analitzem els nivells de diverses adipes/citocines pro i antiinflamatòries en sèrum i secretoma de placa inestable mitjançant assaigs immunoabsorbents lligats a enzims. Es van detectar alguns canvis proteòmics en els secretomes de les plaques ateroscleròtiques carotídies (CAP) en comparació amb les artèries mamàries no ateroscleròtiques (AMNE). Les proteïnes augmentades als secretomes de CAP van ser la defensina 1 de neutròfils, l'apolipoproteïna E, la clusterina i la glicoproteïna alfa-2 de zinc. Pel que fa als nivells sèrics de visfatina, no hi va haver diferències entre els grups de CAP inestable i AMNE.
- English
Arteriosclerosis, atheroma plaque formation, is a highly prevalent pathological process that starts in childhood and progresses over the years. Atherosclerosis is the underlying cause of important clinical manifestations of vascular diseases such as myocardial infarction, stroke or peripheral arterial occlusive disease.
With the increasing incidence of atherothrombosis and the risk of carotid atheroma plaque rupture, the search for novel therapeutic approaches and early biomarkers is a priority. Moreover, new biomarkers should have the potential to improve risk stratification, diagnosis or treatment. . The carotid artery is the artery most frequently used in studies on atherosclerosis. The reasons are the significant accessibility and great quantity of material obtained from endarterectomy Objectives of this thesis are to identify potential candidate biomarkers for carotid atherosclerosis in secretome samples of patients with unstable carotid plaque and to study the possible role of some adipo/cytokines in the stability of atherosclerotic plaques. We also analysed the levels of several pro- and anti-inflammatory adipo/cytokines in serum and unstable plaque secretome according to the presence of obesity, arterial hypertension, diabetes mellitus, dyslipidaemia and smoker status. We analyzed the protein secretion profile of carotid atherosclerotic plaque and non-atherosclerotic mammary secretomes. To evaluate their potential use as atherosclerotic biomarkers, we also studied the functional pathways in which these secreted proteins are involved.
We performed two different studies of human carotid atherosclerotic plaques (CAPs) obtained from patients (men, n = 12) who underwent carotid endarterectomy at the Angiology and Vascular Surgery Unit of the Hospital Universitari Joan XXIII (Tarragona, Spain). Patients with cerebrovascular ischemia and internal carotid artery stenosis >75% were included, diagnosed by color Doppler assisted duplex imaging and arteriography. The CAP diagnosis was made by an experienced pathologist following the American Heart Association (AHA) guidelines . Mammary arteries were used as non-atherosclerotic control arteries (MA).
Segments of mammary arteries (men, n = 10) were obtained during coronary revascularization surgery at the Cardiovascular Surgery Department of the Germans Trias i Pujol Hospital (Badalona, Spain). Patients who had an acute illness, acute or chronic inflammatory or infective diseases, or malignant neoplastic disease were excluded .Blood samples were obtained from each individual immediately before surgery and after overnight fasting. Serum was obtained by standard protocols and preserved at 80 °C until use In a first study we performed proteomic quantitative analysis and in a second, we analysed the levels of several pro- and anti-inflammatory adipo/cytokines in serum and unstable plaque secretom by enzyme-linked immunosorbent assays (ELISA) following the manufacturer’s instructions. We obtained substudy results with statistical analysis of results from a 2nd study.
In a first study we concluded that iTRAQ labelling spectrometry, some proteomic changes were detected in the secretomes of carotid atherosclerotic plaques compared with non atherosclerotic mammary arteries. Some proteins involved in focal adhesion, oxidative stress, inflammation, and endothelial dysfunction, among others, were differentially identified in the secretomes of carotid atherosclerotic plaques. The increased proteins in CAP secretomes were neutrophil defensin 1, apolipoprotein E, clusterin and zinc-alpha-2-glycoprotein. Prospective studies are needed to confirm which profile of secreted proteins could be useful targets for diagnosing and treating carotid atherosclerosis.
In a second study we detected that of all the adipo/cytokines analysed in secretomes, visfatin was the only cytokine that was increased in unstable carotid artery plaques compared with nonatherosclerotic mammary artery secretomes. Regarding visfatin serum levels, there were no differences between the unstable carotid atherosclerotic plaque and nonatherosclerotic mammary artery groups. However, visfatin circulating levels in patients with atherosclerosis were increased compared with those in a serum cohort of healthy subjects, suggesting a possible role of visfatin in atherogenic inflammatory diseases. The exact and precise mechanisms underlying the biological effects of visfatin require further investigation.
Despite the fact that the patients in substudy 3 had a very high cardiovascular risk and were treated with lipid-lowering drugs, only 55.5% of them reached the recommended LDL cholesterol levels according to the clinical guidelines. Circulating adipo/cytokines did not distinguish patients with carotid atherosclerosis and different clinical manifestations of metabolic syndrome. Low adiponectin secretome levels were found in patients with carotid atherosclerosis and obesity.
- español
