Profile of Macrophages in Human Abdominal Aortic Aneurysms: A Transcriptomic, Proteomic, and Antibody Protein Array Study

Abstract

Abdominal aortic aneurysms (AAA) are defined by an increased aortic diameter and characterized by impairment of the extracellular matrix, macrophages infiltration and decreased density of smooth muscle cells. Our aim is to identify the key molecules involved in the pathogenesis of AAAs. This study investigated transcriptomic and proteomic profiles of macrophages from AAA patients (>50 mm aortic diameter) (n = 24) and peripheral arterial occlusion (PAO) patients without AAA detected (n = 18), who both needed a surgery. An antibody protein microarray, generated by printing antibodies onto membranes against proteins selected from the transcriptomic and proteomic analysis, was performed to validate the proteins differentially expressed specifically in macrophages and plasma from the same patients. We found a restricted number of proteins differentially expressed between AAA and PAO patients: TIMP-3, ADAMTS5, and ADAMTS8 that differ significantly in plasma of AAA patients compared to PAO patients, as found in the macrophages. In contrast to plasma MMP-9, soluble glycoprotein V (sGPV) and plasmin-antiplasmin complex levels, plasma TIMP-3 levels were not correlated to AAA size but interestingly correlated to sGPV, a platelet activation marker. Combining transcriptomic and proteomic is a valid approach to identify diseases causing proteins and potential biomarkers.

Transcriptomic and proteomic profiles of vascular cells involved in human abdominal aortic aneurysm

“…All the PAO patients had undergone recent vascular imaging of the aorta and lower limbs before surgery: none had AAAs or iliac, femoral or popliteal aneurysms. The groups did not differ concerning their cardiovascular treatment (Table 2, Lamblin et al, 2010). Before surgery, fewer than two thirds had antiplatelet treatment.…”

“…The LILle Anevrysmal Study (LILAS) was a case-control study that enrolled 42 men, either AAA (case patients) or peripheral arterial occlusion (PAO) (control patients) who needed a vascular surgery or endovascular treatment at the Lille University Hospital Centre (Lille, France) (Lamblin et al, 2010). The ethics committee of the Lille University Hospital Center (France) approved the study (CCPPRB n° CP03/47 of 06 May 2003), and each patient provided written informed consent.…”

“…Specific oligonucleotides for 137 genes corresponding to 24 matrix metalloproteinases (MMP), 4 tissue inhibitors of metalloproteinases (TIMP), 20 ADAMs (a disintegrin and metalloproteinase), 20 ADAMTS (ADAMS with thrombospondin motifs), 65 other proteases and 4 control genes (hydroxymethylbilane synthase, beta-actin, NADPH oxidase 1, and Glyceraldehyde-3-phosphate dehydrogenase (GADPH)) were designed with OLIGOMER software (Mediagen, France) as previously described (Lamblin et al, 2010). Data were analyzed in comparison to a reference RNA, mixture from brain, colon, heart, placenta and testis, chosen for the expression of each protease and anti-protease in at least one of these tissues.…”

“…Interestingly, TIMP-3 was shown to be modulated in the two cell types involved in www.intechopen.com AAA, SMCs and macrophages and also in the plasma of the same patients, as shown with the overexpression of TIMP-3 in AAA tissue compared to aortic occlusive disease but they did not measure plasma TIMP-3 levels of the same patients as we did (Carell et al, 2002). The availability of TIMP assays allowed us to confirm the increased plasma levels of TIMP-3 in AAA patients (Lamblin et al, 2010 Macrophages and SMC analysis were performed respectively from 16 and 14 AAA and 16 and 11 PAO samples; Plasma analysis was performed from 24 AAA and 18 PAO samples. * Due to the limited number of SMC samples tested, we did not calculated SD for PAO patient and gave the value from 2 independent experiments performed.…”

Inactivation of PI3Kδ Induces Vascular Injury and Promotes Aneurysm Development by Upregulating the AP-1/MMP-12 Pathway in Macrophages

“…Monocytes/ macrophages infiltrate the vessel wall and release proteases, including elastase (matrix metalloproteinase-12 ) and metalloproteinases that compromise the integrity of the vascular wall through degradation of the extracellular matrix (ECM). Monocytes/macrophages also secrete inflammatory cytokines in the media and adventitia of aneurysmatic vessels, such as tumor necrosis factor (TNF)-α, interferon-γ, interleukin-1β, [9][10][11] and interleukin-6, 12 which are reported significantly increased in patients with AAA. As a central role in AAA, macrophages may be classically activated by tolllike receptor (TLR) ligands.…”