Proteomic and histopathologic profiling reveal molecular features and clinical biomarkers of coronary atherosclerosis

Introduction: The key molecular mechanisms associated with human atherosclerotic plaque progression, atherosclerotic lesion initiation and plaque destabilization are not fully understood. Hypothesis: Different stages of atherosclerosis (AS) progression have distinct pathological processes. Some key proteins induce AS progression. Methods: We employed 20 coronary artery samples among 4 explanted hearts from coronary heart disease patients. Each heart contains stages of unaffected control (NC), pathological intimal thickening (PIT), fibroatheroma (FA), transition to thin-cap fibroatheroma (TCFA) and ruptured plaque (RP), which were validated by hematoxylin and eosin (H&E) staining. We performed quantitative proteomics in each of the samples based on tandem-mass-tags and liquid chromatography-tandem mass spectrometry. Furthermore, we measured proteins in human coronary artery tissue via immunofluorescence. Results: Proteomics revealed that extracellular matrix (ECM) organization was the most significantly enriched pathway in PIT. FA exhibited an enrichment of immunologic processes, such as neutrophil degranulation, myeloid leukocyte activation and innate immune response. Molecular profiles of TCFA and RP are similar, with complement and coagulation cascades pathway as a hallmark of these stages. Clustering analysis of DEPs displayed 4 clusters, notably cluster 2 had a precipitous elevation from FA to TCFA. Within this cluster, we identified a high-rank molecule: macrophage-capping protein (CAPG). Immunohistochemistry and immunofluorescence revealed that CAPG is highly elevated in plaque compared to NC and is specifically expressed in the plaque. Conclusions: ECM organization was continuously dysregulated in AS progression. Immune response might initiate the atherosclerotic plaque. Complement and coagulation cascades pathway was a hallmark of TCFA and RP. CAPG might be a therapeutic target for AS progression.

Accurate evaluation of the axillary lymph node (ALN) status is needed for determining the treatment protocol for breast cancer (BC). The value of magnetic resonance imaging (MRI)-based tumor heterogeneity in assessing ALN metastasis in BC is unclear. To assess the value of deep learning (DL)-derived kinetic heterogeneity parameters based on BC dynamic contrast-enhanced (DCE)-MRI to infer the ALN status. Retrospective. 1256/539/153/115 patients in the training cohort, internal validation cohort, and external validation cohorts I and II, respectively. 1.5 T/3.0 T, non-contrast T1-weighted spin-echo sequence imaging (T1WI), DCE-T1WI, and diffusion-weighted imaging. Clinical pathological and MRI semantic features were obtained by reviewing histopathology and MRI reports. The segmentation of the tumor lesion on the first phase of T1WI DCE-MRI images was applied to other phases after registration. A DL architecture termed convolutional recurrent neural network (ConvRNN) was developed to generate the KHimage (kinetic heterogeneity of DCE-MRI image) score that indicated the ALN status in patients with BC. The model was trained and optimized on training and internal validation cohorts, tested on two external validation cohorts. We compared ConvRNN model with other 10 models and the subgroup analyses of tumor size, magnetic field strength, and molecular subtype were also evaluated. Chi-squared, Fisher's exact, Student's t, Mann-Whitney U tests, and receiver operating characteristics (ROC) analysis were performed. P < 0.05 was considered significant. The ConvRNN model achieved area under the curve (AUC) of 0.802 in the internal validation cohort and 0.785-0.806 in the external validation cohorts. The ConvRNN model could well evaluate the ALN status of the four molecular subtypes (AUC = 0.685-0.868). The patients with larger tumor sizes (>5 cm) were more susceptible to ALN metastasis with KHimage scores of 0.527-0.827. A ConvRNN model outperformed traditional models for determining the ALN status in patients with BC. 3 TECHNICAL EFFICACY: Stage 2.

One of the main risk factors for cerebral ischemic events is atherosclerotic disease of the internal carotid artery (ICA). Nowadays, increasing attention is being paid to the relationship between the morphological features of atherosclerotic plaque and the occurrence of stroke. Several studies have demonstrated that the presence of specific vulnerable plaque types, with a large lipid core and thin fibrous cap, can be used as an independent risk predictor of cerebral ischemic events. The present study is an attempt to develop the method of plaque surface morphology assessment presented by de Weert et al. by correlating the results of Dual Source Computed Tomography (DSCT) with those from intravascular ultrasound virtual histology (IVUS-VH). A group of 30 symptomatic patients (13 men and 17 women; 72 ± 9 years) with ICA stenosis suspected on the basis of ultrasound imaging (US) and confirmed to be above 70% in DSCT underwent intravascular ultrasound (IVUS) imaging. The results of DSCT were categorized according to the de Weert classification. There were 13 cases (43%) with smooth wall surfaces, 10 cases (33%) with discreet wall irregularities, and seven cases (23%) with incursions of contrast, indicating the presence of ulceration. In the IVUS-VH examinations, 4 out of 30 cases (13%) were identified as having adaptive intimal thickening (AIT), 4 (13%) as showing pathological intimal thickening (PIT), 6 (20%) with fibroatheromas (FA), six (20%) with fibrocalcific plaque (FCa), and 10 (33%) as having thin-cap fibroatheroma (TCFA), which is high-risk plaque. Comparing the above results showed that all the patients with confirmed wall ulceration in DSCT were characterized as having high-risk plaque in IVUS-VH. Using DSCT with the de Weert classification of plaque surface morphology makes reliable detection of ulcerations possible; therefore, this could become a significant new technique to improve current imaging protocols for patients with a high risk of ischemic cerebrovascular events.

Pathology of high risk atherosclerotic plaque provides the basis for understanding the imaging and treatment of atherosclerosis. The earliest vascular change described microscopically are adaptive intimal thickening and fatty streaks, whereas pathologic intimal thickening are the first of the progressive plaques subtypes. Fibroatheromas are characterized by an acellular necrotic core, accumulated cellular debris and cholesterol monohydrate, and a lack of extracellular matrix. The development of the necrotic core is believed to originate from apoptotic macrophages. Thinning of the fibrous cap leads to plaques vulnerable to rupture, or thin-cap fibroatheromas. Overlying thrombosis can arise from one of several mechanisms including ruptures, erosion, or calcified nodules. Calcium within atherosclerosis is a common imaging target which increases with lesion progression and is present in greatest frequent in healed plaque ruptures and fibrous plaques. Thin cap fibroatheromas most frequently contain speckled calcification but may show heavily calcified areas or an absence of calcification. which is not very useful in diagnosing these lesions by calcium-based imaging. Coronary lesions with thrombi in the absence of rupture (erosions) exclusively show stippled or no calcification. Rupture in the absence of calcification is rare. In contrast, diffuse calcification is almost always associated with healed ruptures.

Thin-cap fibroatheromas (TCFA) are prone to plaque rupture and thrombosis. Intravascular ultrasound (IVUS) virtual histology (VH) assesses plaque composition and lesion morphology in vivo. Methods &amp; Results: We used serial (baseline and follow-up @11 mos) VH-IVUS to study non-culprit plaque morphology in 221 lesions (plaque burden &gt;40%) in 106 pts. Lesions were classified into 4 types based on plaque composition; pathological intimal thickening (PIT), thin-capped fibroatheroma (TCFA), thick-capped fibroatheroma (ThCFA), fibrotic/fibrocalcific. At baseline, 21 lesions were TCFAs (confluent necrotic core contacting to the lumen). Overall during follow-up (Figure ), 16/21 (76%) TCFAs healed: 13 became ThCFAs, 2 TCFAs became PIT, 1 TCFA became fibrotic, and 5 TCFAs (24%) remains unchanged although the location of the necrotic core in contact with the lumen shifted axially. Compared to TCFAs that healed, TCFAs that remained TCFAs were more often proximal in location (distance from coronary ostium to the lesion of 19±6 vs. 41±22mm, respectively, p=0.037) and had larger lumen area (9.9±3.1 vs. 6.9±1.8 mm2, p=0.013), vessel area (22.8±6.4 vs. 15.3±2.6 mm2, p=0.001), plaque area (12.9±4.9 vs. 8.4±1.7 mm2, p=0.005), calcium area (1.3±0.6 vs. 0.6±0.3 mm2, p=0.014), and necrotic core area (2.6±1.0 vs. 1.6±0.7 mm2, p=0.015). In addition, 6 new TCFAs developed; these 6 late-developing TCFAs had the appearance of PIT at baseline (Figure ). Conclusion: Although most TCFAs seem to stabilize or heal during 12 mos follow-up, proximal TCFAs in larger vessels with more plaque and calcium and a larger necrotic core appear to heal less often and new TCFAs can develop.

The term “ P athologic I ntimal T hickening” (PIT) was recently introduced to define an early stage of atherosclerosis described in human coronary lesions found at autopsies of sudden death victims.1 This descriptive identifier is based on the AHA type III (intermediate) lesion and, as originally presented by Stary and collegues, it’s believed to be the morphological and chemical bridge to more advanced plaques.2 The precise histological features and clinical relevance of PIT remains unsettled, and use of the term is still far from widespread. In short, PIT identifies a lesion with an extracellular lipid pool with intimal smooth muscle cell loss typically adjacent to the medial wall in addition to varying degrees of macrophage infiltration near the lumen. These morphological features indicate a progressive lesion in the earlier stages of atherosclerosis, although there is yet the presence of a necrotic core. As recently studied by Nakashima and colleagues in this issue of Arteriosclerosis, Thrombosis, and Vascular Biology , it may provide a key in settling the chicken-versus-egg debate of atherosclerotic plaque progression: does lipid come first, or do macrophages?3 Is PIT the precursor lesion of fibroatheroma? The study in this issue uses 3-dimensional histology to attempt to address some of these issues, and, in doing so, may raise as many questions as it answers. See page 1159 Although there are many detailed autopsy studies describing various lesion morphologies, little is known how human atherosclerosis progresses from early to more advanced plaques, marked by the formation of a necrotic core. This important question remains, in part, from a lack of direct experimental testing in prospective models of human disease. Moreover, potentially relevant finding in animals are difficult to associate with humans because the pathologic change of atherosclerosis in man cannot be definitely equated with animals. Although …

To develop and evaluate nomograms incorporating clinical features and quantitative CT parameters for the preoperative prediction of microvascular invasion (MVI) and pathological grading in patients with hepatocellular carcinoma (HCC). This retrospective multicenter study involved 684 consecutive patients with pathologically confirmed HCC. In this context, 553 patients from Center 1 were randomly split into training (70%) and internal validation (30%) cohorts, and 131 patients from Center 2 served as an external validation cohort. Predictive factors for MVI-positive and high-grade HCC were identified through univariate and multivariate logistic regression. Two nomograms combining clinical factors and CT quantitative parameters were developed. For MVI prediction, multivariate analysis identified HBV infection, an AFP concentration > 20 ng/mL, a larger tumor diameter, a higher arterial absolute enhancement value (AAEV), and a lower portal relative enhancement ratio (PRER) as independent predictors. The nomogram achieved area under the curve (AUC) values of 0.769 (95% CI: 0.720-0.818), 0.771 (0.692-0.850), and 0.760 (0.648-0.872) in the training, internal validation, and external validation cohorts, respectively. For pathological grade prediction, younger age, an AFP concentration > 20 ng/mL, a larger tumor diameter, and a lower PRER were independent predictors. The corresponding nomograms had AUCs of 0.696 (0.639-0.752), 0.644 (0.547-0.741), and 0.768 (0.681-0.856) across cohorts. Both nomograms demonstrated excellent calibration and significant clinical utility for decision curve analysis. Nomograms integrating clinical features and quantitative CT parameters facilitate accurate preoperative prediction of MVI status and pathological grading in HCC patients, demonstrating strong potential for clinical implementation. Question Predicting microvascular invasion (MVI) and pathological grading preoperatively in hepatocellular carcinoma (HCC) patients using a clinically practical and cost-effective method remains challenging. Findings Quantitative parameters derived from contrast-enhanced CT demonstrated the ability to predict MVI and pathological grade, with validated models demonstrating robust diagnostic performance and clinical applicability. Clinical relevance Nomograms, which integrate clinical features and quantitative CT parameters, represent a clinically applicable tool for the preoperative prediction of MVI status and pathological grading in HCC patients, facilitating personalized treatment planning and optimized patient management.

Longitudinal changes in tumor morphology and body composition following neoadjuvant chemotherapy predict progression-free survival in adenocarcinoma of the esophagogastric junction: A multi-center study.

This study evaluated whether optical frequency domain imaging (OFDI) accurately distinguish between fibroatheroma (FA) and pathological intimal thickening (PIT) compared with histopathology. A total of 631 histological cross-sections from 14 autopsy hearts were analyzed for the comparison between OFDI and histological images. Of those, 190 (30%) sections were diagnosed with PIT and 120 (19%) with FA. The OFDI signal attenuation rate was calculated from an exponential. The lipid length was measured longitudinally by detection of sequential OFDI frames within a plaque segment containing lipids. The lipid arc was measured with a protractor centered in the center of the lumen. The fibrous cap thickness was defined as the minimum thickness of the signal rich band overlying PIT and FA. There was no significant difference in the OFDI signal attenuation rate between FA and PIT (3.09 ± 1.04 versus 2.79 ± 1.20, p = 0.13). However, the lipid length was significantly longer, the maximum lipid arc was significantly larger, and the fibrous cap thickness was significantly thinner in FA than in PIT (7.5 [4.3-10.3] mm versus 4.3 [2.7-5.8] mm, p < 0.0001, 125 [101-174]° versus 96 [74-131]°, p < 0.0001, and 220 [167-280] µm versus 260 [190-332] µm, p = 0.019). This study revealed OFDI may have the potential capability for discriminating FA from PIT based on the longitudinal and circumferential extent of lipid plaque, although the OFDI signal attenuation rate was similar between FA and PIT.

Dynamic Nature of Nonculprit Coronary Artery Lesion Morphology in STEMI: A Serial IVUS Analysis From the HORIZONS-AMI Trial

Coronary artery disease (CAD) represents a global leading cause of death especially for middle- and high-income countries. In 2012, all-cause deaths were estimated as 56 million worldwide with 17.5 million deaths attributed to cardiovascular disease (CVD). The initial concept of atherosclerosis progression and plaque phenotypic change was established by Dr. Velican in the early 1980s who focused on morphological descriptions of fatty streak lesions to advanced fibroatheromatous plaques complicated by haemorrhage, calcification, ulceration and thrombosis. Numeric American Heart Association (AHA) lesion types I-IV were essentially replaced by descriptive terminology to include adaptive intimal thickening, intimal xanthoma, pathologic intimal thickening (PIT) and fibroatheroma (FA) with recognised progressive lesion morphology of early and late FAs. Intimal thickening in the absence of atherosclerosis is considered the earliest vascular change consisting primarily of smooth muscle cells (SMCs) and extracellular matrix. PIT is considered the earliest of progressive change.

Progression of coronary artery disease: Cinearteriographic and clinical observations in medically and surgically treated patients

Thin-cap fibroatheroma (TCFA) is the most common type of vulnerable plaque and is the precursor of plaque rupture. However, rupture of a TCFA is not the only mechanism underlying thrombus formation or acute coronary syndrome. Although statin therapy changes the composition of coronary artery plaques, the effects of statins, particularly different types of statins, on plaque phenotype have not been fully examined. This study compared the effects of pitavastatin versus pravastatin on coronary artery plaque phenotype assessed by virtual histology (VH) intravascular ultrasound (IVUS) in patients with angina pectoris (AP). Coronary atherosclerosis in nonculprit lesions was evaluated using VH-IVUS at baseline and 8 months after statin therapy; analyzable IVUS data were obtained from 83 patients with stable AP (39 patients treated with pitavastatin and 44 with pravastatin) and 36 patients with unstable AP (19 patients treated with pitavastatin and 17 with pravastatin). Pitavastatin had a strong effect on reducing pathologic intimal thickening (PIT), especially in patients with unstable AP, but had no impact on VH-TCFA or fibroatheroma (FA). By contrast, pravastatin had weak effects on reducing PIT, VH-TCFA, or FA. Increases in the number of calcified plaques were observed for both statins. In conclusion, pitavastatin and pravastatin changed coronary artery plaque phenotype as assessed by VH-IVUS in patients with AP. However, the effects of these statins on coronary artery plaque phenotype were different.

Background ; Few studies have investigated the association between biomarkers and in vivo plaque vulnerability in patients with stable coronary artery disease (CAD). Methods ; We examined two or three vessels of 55 stable CAD patients with combined use of virtual histology intravascular ultrasound (VH-IVUS) and optical coherence tomography (OCT), and diagnosed thin-cap fibroatheroma (TCFA). The definition of TCFA was described as follow: necrotic core rich lesion (%necrotic-core&gt;10%) without evident overlying fibrous component and %plaque-volume&gt;40% by VH-IVUS analysis, and the thinnest fibrous-cap thickness &lt;65μm by OCT measurement. The patients were divided into two groups, the patients with TCFA and without TCFA, and compared their clinical characteristics, laboratory data and new atherosclerotic biomarkers (adiponectin, oxidative stress marker) between these two groups. Results ; In 55 patients, we could observe 126 vessels (2.29±0.46 vessel/patient) with both modality and we identified 37 TCFA in them. At least one TCFA was identified in 24 patients (43.6%) and not in 31 patients. There were no differences in the clinical characteristics between the two groups. However, a greater number of patients with TCFA had a past history of ACS than those without TCFA (P=0.04). As for laboratory data, there were no differences in lipid and sugar profiles and inflammatory markers, but patients with TCFA had lower plasma adiponectin levels and higher urine- 8-isoprostane/creatinine levels, which is considered as reliable in vivo oxidative stress marker, than patients without TCFA (p=0.0002, P=0.04, respectively). Multivariate logistic regression analysis revealed the most powerful indicative factor of TCFA prevalence in stable CAD was plasma adiponectin levels (P=0.001). The receiver-operating characteristic curve analysis identified plasma adiponectin &lt;6.8μg/ml (Area under curve = 0.80, 95% confidence interval = 0.67 to 0.90, P=0.0001, sensitivity 70.8%, specificity 80.6%) as the optimal cut-off point for prediction of the TCFA prevalence in stable CAD. Conclusion ; Plasma adiponectin levels and urine-8-isoprostane may be one of the tools that help to stratify vulnerable patients into risk categories in patients with stable CAD.

Introduction: Current methods to identify thin-cap fibroatheroma (TCFA) with optical coherence tomography (OCT) are qualitative. While two mechanisms for the appearance of false TCFA have been identified and include macrophages that cause superficial shadowing and tangential light dropout, we report three new mechanisms for the appearance of false TCFAs in OCT images. Methods: 13 coronary arteries from 8 human cadaver hearts (6 male, 2 female, aged 63 ± 12 years) were imaged with OCT. OCT TCFAs were identified independently by three expert OCT readers and defined by a bright homogenous fibrous cap overlying a diffusely shadowed signal-poor region. Analysis was performed to confirm the identification of TCFA or to identify the histologic origin of the OCT TCFA appearance. Arc length of the signal-poor region of each OCT TCFA was also measured. Results: 38 OCT TCFA were identified, of which only 5 were verified with histology. Measurements of arc length and thickness of the cap are summarized in the table. Of the 33 false TCFA, three morphologic plaque types were identified: thick-cap fibroatheroma (TkFA; N=18), pathologic intimal thickening (PIT; N=10), and fibrocalcific (FC; N=5). New mechanisms for the signal-poor regions that appeared as lipid cores included loose connective tissue (N=7), neovasculature (N=3), and proteoglycans (N=4). Superficial shadowing of macrophages caused the appearance of TCFA in 5 cases. In the 18 TkFA cases, OCT underestimated the thickness of the fibrous cap (45.2 ± 4.0 μm by OCT compared to 197.5 ± 18.8 μm by histology). Conclusion: Using an arc length of greater than 90 degrees will aid in differentiating true from false TCFA. Additionally, due to incomplete light penetration, TkFA can be mistaken for TCFA. These results demonstrate that caution should be used when diagnosing vulnerable plaque with OCT.