Access

Format

Author / Creator

Subject

Recently added

Eresources

Language

Place of Publication

Dates

Location

Thoracic aortic aneurysm and dissection (TAAD) are complex disease states with high morbidity and mortality that pose significant challenges to early diagnosis. Patients with an aneurysm are asymptomatic and typically present to the emergency department only after the development of a dissection. The extracellular matrix (ECM) plays a crucial role in regulating the aortic structure and function. The histopathologic hallmark termed medial degeneration is characterised by smooth muscle cell (SMC) loss, the degradation of elastic and collagen fibres and proteoglycan (PG) accumulation. Covalently attached to the protein core of PGs are a number of glycosaminoglycan chains, negatively charged molecules that provide flexibility, compressibility, and viscoelasticity to the aorta. PG pooling in the media can produce discontinuities in the aortic wall leading to increased local stress. The accumulation of PGs is likely due to an imbalance between their synthesis by SMCs and decreased proteolysis by A Disintegrin-like and Metalloproteinase with Thrombospondin motifs (ADAMTS) proteoglycanases in the ECM. Mouse models of TAAD indicated that these proteases exert a crucial, albeit complex and not fully elucidated, role in this disease. This has led to a mounting interest in utilising ADAMTS proteoglycanases as biomarkers of TAAD. In this review, we discuss the role of ADAMTSs in thoracic aortic disease and their potential use in facilitating the clinical diagnosis of TAAD and disease progression.

International journal of molecular sciences, 2023-07, Vol.24 (15), p.12135

The Role of ADAMTS Proteoglycanases in Thoracic Aortic Disease

BackgroundThoracic Aortic Aneurysm (TAA) is a disease with significant morbidity and mortality. The aneurysm is asymptomatic, and patients often present to the emergency department after a dissection. A reliable biomarker is required to diagnose the condition early to direct the patient to the appropriate services. TAA is characterised by proteoglycan accumulation which are normally found in the extracellular matrix (ECM) of the medial layer. This may be due to impaired activity of intrinsic proteases A Disintegrin and Metalloproteinase with Thrombospondin Motifs (ADAMTS) 4 and 5. We aimed to find a circulating proteoglycan-based biomarker associated with abnormal material properties of the diseased aortic wall.MethodsADAMTS-generated fragment ‘versikine’, ADAMTS4, ADAMTS5, ADAMTS13 and von Willebrand Factor (vWF) were measured in the plasma of patients with TAA (n=48) and healthy controls (n=23) using Enzyme-Linked Immunosorbent Assays. Patients with connective tissue disease (CTD) and bicuspid aortic valve were excluded from the study. Mann-Whitney U test was used to compare the analyte plasma levels between the groups and Spearman Rank to test for correlation. Receiver-operator characteristics curve analysis was used to generate specificity and sensitivity values. In five patients, explanted TAA samples from surgery were processed to extract proteoglycans using 4M Guanidine HCl and purified by Diethylaminoethyl chromatography. Western Blot was used to qualitatively analyse the fragments.ResultsVersikine plasma concentration was higher in TAA patients compared to controls (34.23±3.83 vs 10.05±1.15 ng/mL, p<0.0001). In TAA patients, versikine concentration was negatively correlated with the aortic diameter (r=-0.3712, p=0.0218) and outer curve thickness (r=-0.3819, p=0.0493). ADAMTS4 plasma concentration was lower in TAA patients compared to controls (11.02±2.88 vs 25.02±5.07 ng/mL, p=0.0114). ADAMTS5, ADAMTS13 and vWF concentrations were similar in both groups. Versikine plasma levels could identify TAA with a sensitivity of 68.1% and specificity of 95.6%. In addition, high levels of versikine were present in the aneurysmal aortic tissue.Abstract BS57 Figure 1Schematic Representation of Thoracic Aortic Aneurysm (TAA) pathogenesis and biomarker panel selection. A) A diagram of the thoracic aneurysm in the ascending aorta where the tunica media layer of the aorta has been zoomed in to show an accumulation of proteoglycans (PG) in the extracellular matrix (ECM) where the smooth muscle cells (SMCs) reside. The wider lumen can be seen in TAA aorta compared to healthy aorta. There is also loss of SMCs in the media layer. B) Normal aorta in healthy adult where a reduced content of proteoglycans (PGs) can be observed compared to A. C) Blood sample taken from patients and healthy controls. D) 96-Well Plate used for all the Enzyme-linked immunosorbent assay (ELISA) experiments run in the lab to measure plasma content of the potential biomarkers. E) The list of the different molecules that were measured to include in the biomarker panel and to analyse their levels compared to the aortic properties in TAA patients.PG=proteoglycan, SMCs=smooth muscle cells, ECM=extracellular matrix, ADAMTS= A Disintegrin and Metalloproteinase with Thrombospondin Motif, vWF= von Willebrand Factor.Abstract BS57 Figure 2Proteoglycan extraction in human aortic tissue. 1) Aortic aneurysm tissue sample obtained straight after aortic aneurysm repair surgery from the ascending aorta. 2) Adventitia was removed due to the thrombi and fatty deposits that make dissection into smaller fragments challenging. We were interested in extracting proteoglycans mainly from the aortic media layer. 3) After tissue was dissected into superior (region further away from the aortic valve) and inferior (region of the tissue closer to the aortic valve), the tissue was further dissected into square 1.5x1.5cm fragments and stored in -80 after freezing with liquid nitrogen. 4) Frozen tissue was homogenized using a mortar and pestle until it looked almost like a powder. 5) 10mL 4M Guanidine Extraction buffer was adder per 1 g of tissue and homogenate was further centrifugated at speed 18000g for 45 minutes. 6) The filtered homogenate was loaded onto the fast protein liquid chromatography (FPLC) using Diethylaminoethyl cellulose (DEAE). Absorbance was read at OD280 and high OD corresponded to fractions 4-6 and 11-19. 2 mL fractions were collected including the starting material (SM), flowthrough (FT), washing (W). 7) 20uL of all the fractions including SM, W, FT were stained with Dimethylmethylene Blue Assay (DMMB) which binds to glycosaminoglycans (GAGs) and showed high GAG content in fractions 15-17, the SM,FT and W. 8) Western blot analysis using anti-Chondroitin Sulphate (CS) primary antibody showed high protein content with molecular weight 250 kilodalton(kDa) in the SM, FT,W and fractions 15-17. Protein with molecular weight 37kDa was present in SM, FT,W and fractions 14-18. Fractions 15-17 were pooled into pool 1 and fractions 14 and 18 were pooled in pool 2 and underwent another western blot analysis using anti-DPEAAE. The neoepitope antibody detects an N-terminal versican fragment generated by ADAMTS5 cleavage at Glu441 -Ala442 bond (versikine). 10 uL of Pool 2 showed presence of versikine. Anti-G1, anti-full length versican and anti-biglycan antibodies did not give any results using 10mL of the pooled fractions.ConclusionVersikine levels are higher in the plasma of TAA patients which may be due to the high turnover of versican in the aortic tissue. Versikine levels correlated with both aortic diameter and outer curve aortic thickness. Lower ADAMTS4 levels suggest reduced proteolysis which may contribute to the accumulation of proteoglycans in the tissue. Versikine has potential to become a reliable biomarker for diagnosing a TAA. Larger trials including CTD and bicuspid valve patients are required to validate versikine as a biomarker and test its’ ability to predict aortic diameter changes over time.Conflict of Interestnone

Heart (British Cardiac Society), 2023, Vol.109 (Suppl 3), p.A285-A287

BS57 Assessing proteoglycans and proteoglycanases as biomarkers for thoracic aortic aneurysms

Non-ST-segment elevation myocardial infarction (NSTEMI) is a leading cause of emergency hospitalization across Europe. This study evaluates the in-hospital and mid-term outcomes of patients who underwent coronary artery bypass graft (CABG) after NSTEMI. A retrospective analysis of all cases who underwent isolated CABG after NSTEMI from September 2017 to September 2022 at our center. Patients were stratified according to in-hospital survival. Patient characteristics, operative details, and procedural complications were compared between those who survived and those who did not. Predictors of in-hospital and mid-term mortality were evaluated using logistic and Cox regression modeling. Kaplan-Meier analysis was used to generate a survival curve for all alive patients at the time of discharge. Among 1,011 patients (median age 64 [56 to 72] years, 852 [84.3%] male), 735 (72.7%) underwent urgent, 239 (23.6%) elective, and 37 (3.7%) emergency CABG. The in-hospital mortality was 1.5% (15/1,011 patients). Those who died were more likely to be New York Heart Association class III/IV, have left ventricular ejection fraction <21%, severe renal impairment, peripheral vascular disease (PVD), or poor mobility. Emergency procedures, preoperative ventilation, inotropic support, and intra-aortic balloon pump (IABP) use were also more prevalent among those who died. Logistic regression modeling revealed new postoperative stroke (odds ratio 22.0, 95% confidence interval 3.6 to 135.5, p = 0.001), preoperative IABP use (11.4; 2.4 to 53.7, p = 0.002), new hemodialysis (9.6; 2.7 to 34.7, p <0.001), PVD (5.6; 1.6 to 20.0, p = 0.008), and poor mobility (odds ratio 4.8, 95% confidence interval 1.3 to 18.2, p = 0.022) as independent predictors of in-hospital mortality. In conclusion, new postoperative stroke, preoperative IABP use, new hemodialysis, PVD, and poor mobility are independent predictors of mortality in patients with NSTEMI who underwent isolated CABG.
[Display omitted]

The American journal of cardiology, 2024-07, Vol.223, p.165-173

Outcomes of Patients Presenting With Non-ST Elevation Myocardial Infarction Who Underwent Surgical Revascularization

ABSTRACT
Proteoglycans and their proteolytic fragments diffuse into biological fluids such as plasma, serum, urine, or synovial fluid, where they can be detected by antibodies or mass‐spectrometry. Neopeptides generated by the proteolysis of proteoglycans are recognized by specific neoepitope antibodies and can act as a proxy for the activity of certain proteases. Proteoglycan and proteoglycan fragments can be potentially used as prognostic, diagnostic, or theragnostic biomarkers for several diseases characterized by dysregulated extracellular matrix remodeling such as osteoarthritis, rheumatoid arthritis, atherosclerosis, thoracic aortic aneurysms, central nervous system disorders, viral infections, and cancer. Here, we review the main mechanisms accounting for the presence of soluble proteoglycans and their fragments in biological fluids, their potential application as diagnostic, prognostic, or theragnostic biomarkers, and highlight challenges and opportunities ahead of their clinical translation.

Proteoglycan research, 2024-10, Vol.2 (4), p.e70011-n/a

Soluble Proteoglycans and Proteoglycan Fragments as Biomarkers of Pathological Extracellular Matrix Remodeling

Export record to excel

Export this record

Export record to excel

The Role of ADAMTS Proteoglycanases in Thoracic Aortic Disease

Export this record

Export record to excel

BS57 Assessing proteoglycans and proteoglycanases as biomarkers for thoracic aortic aneurysms

Export this record

Export record to excel

Outcomes of Patients Presenting With Non-ST Elevation Myocardial Infarction Who Underwent Surgical Revascularization

Export this record

Export record to excel

Soluble Proteoglycans and Proteoglycan Fragments as Biomarkers of Pathological Extracellular Matrix Remodeling

Refine your search