Histological Validation Research Articles

Histological Validation of MRI: A Review of Challenges in Registration of Imaging and Whole-Mount Histopathology.

Rigorous validation with ground truth information such as histology is needed to reliably assess the current and potential value of MRI techniques to characterize tissue and identify disease-related tissue alterations. Commonly used methods that aim to directly correlate histology and MRI data generally fall short of this goal due to spatial errors that preclude direct matching. Errors result from tissue deformation, differences in spatial resolution and slice thickness, non-coplanar and/or nonintersecting plane orientations, and different image contrast mechanisms. Some of these problems arise from limitations in standard protocols for clinical tissue processing and histology-based pathology reporting, and to some extent can be addressed by modifications to standard protocols without compromising the clinical process. Typical modifications include ex vivo specimen MRI, block-face photography, addition of fiducial markers, and 3D printed molds to constrain tissue deformation and guide sectioning. This review summarizes the advantages and limitations of MRI validation techniques based on coregistration of MRI with whole-mount histology of tissue specimens. LEVEL OF EVIDENCE: 4 TECHNICAL EFFICACY STAGE: 1.

The National Endoscopy Database (NED) Automated Performance Reports to Improve Quality Outcomes Trial (APRIQOT) randomized controlled trial design.

Background and study aims Colonoscopists with low polyp detection have higher post colonoscopy colorectal cancer incidence and mortality rates. The United Kingdom’s National Endoscopy Database (NED) automatically captures patient level data in real time and provides endoscopy key performance indicators (KPI) at a national, endoscopy center, and individual level. Using an electronic behavior change intervention, the primary objective of this study is to assess if automated feedback of endoscopist and endoscopy center-level optimal procedure-adjusted detection KPI (opadKPI) improves polyp detection performance. Methods This multicenter, prospective, cluster-randomized controlled trial is randomizing NHS endoscopy centres to either intervention or control. The intervention is targeted at independent colonoscopists and each center’s endoscopy lead. The intervention reports are evidence-based from endoscopist qualitative interviews and informed by psychological theories of behavior. NED automatically creates monthly reports providing an opadKPI, using mean number of polyps, and an action plan. The primary outcome is opadKPI comparing endoscopists in intervention and control centers at 9 months. Secondary outcomes include other KPI and proximal detection measures at 9 and 12 months. A nested histological validation study will correlate opadKPI to adenoma detection rate at the center level. A cost-effectiveness and budget impact analysis will be undertaken. Conclusion If the intervention is efficacious and cost-effective, we will showcase the potential of this learning health system, which can be implemented at local and national levels to improve colonoscopy quality, and demonstrate that an automated system that collects, analyses, and disseminates real-time clinical data can deliver evidence- and theory-informed feedback.

Histological validation of simultaneous non-contrast angiography and intraplaque hemorrhage imaging (SNAP) for characterizing carotid intraplaque hemorrhage.

This study sought to validate the performance of simultaneous non-contrast angiography and intraplaque hemorrhage (SNAP) imaging in characterizing carotid IPH by histology. Thirty-five patients with carotid atherosclerotic disease (symptomatic 50-70% stenosis or > 70% stenosis) scheduled for carotid endarterectomy underwent 3.0-T carotid MR imaging by acquiring SNAP and magnetization-prepared rapid acquisition gradient-echo (MP-RAGE) sequences. Presence and area of IPH were separately evaluated on SNAP and MP-RAGE images. Presence and area of IPH were also assessed on histology. Agreement between SNAP/MP-RAGE and histology was determined in identify and quantify IPH using Cohen kappa, Spearman correlation, and Bland-Altman analyses. Of all 35 patients (mean age: 63.1 ± 8.8 years; 27 males), 128 slices with successful registration were eligible for analysis. The accuracy, sensitivity, specificity, and positive and negative predictive values were 86.7%, 85%, 89.6%, 93.2%, and 78.2% for SNAP, and76.6%, 75%, 79.2%, 85.7%, and 65.5% for MP-RAGE in identification of IPH, respectively. In identification of IPH, the kappa value between SNAP and histology and between MP-RAGE and histology was 0.725 and 0.520, respectively. The correlation between SNAP and histology (r = 0.805, p < 0.001) was stronger than that between MP-RAGE and histology (r = 0.637, p < 0.001) in measuring IPH area. Bland-Altman analysis showed that, in measuring IPH area, the bias of SNAP (1.4 mm2, 95% CI: - 0.016 to 2.883) was smaller than that of MP-RAGE (1.7 mm2, 95% CI: - 0.039 to 3.430) compared with histology. This validation study by histology demonstrates that SNAP sequence better identifies and quantifies carotid intraplaque hemorrhage compared with traditional MP-RAGE sequence. • SNAP imaging showed better agreement with histology compared with MP-RAGE imaging, especially for the IPHs with small size. • SNAP sequence is a more effective tool to identify and quantify carotid IPH than traditional sequence of MP-RAGE that can help clinicians to optimizing the treatment strategy. • The plaque components of rich lipid pools or loose matrix and chronic/old IPH (cholesterol crystals) can lead to false positive and false negative results in SNAP and MP-RAGE imaging for identifying IPH.

Contrast-enhanced micro-CT to assess dental pulp tissue debridement in root canals of extracted teeth: a series of cascading experiments towards method validation.

To validate a new method for the evaluation of pulp tissue debridement in the root canals of extracted teeth using an impregnation protocol involving potassium triiodide, a radiocontrast solution known as Lugol's, combined with micro-computed tomographic (micro-CT) imaging. The impact of NaOCl on the radiopacity of Lugol's solution was assessed using a two-fold dilution series of Lugol in distilled water and 5.25% NaOCl, which were then pipetted into transparent dishes and radiographed. To verify the influence of Lugol on the proteolytic effect of NaOCl, a dissolution test was performed using fresh bovine meat. Ten slices did not undergo any tissue processing, whilst twenty slices were fixed in formaldehyde for 24h. After that, 10 of them were immersed in Lugol for another 24h. Then, all specimens were placed in NaOCl and the time required for a complete tissue dissolution was recorded. For the last experiments (histological validation and micro-CT assessment), 8 extracted mandibular premolars with formerly vital pulps were immersed in buffered formalin, scanned in a micro-CT device, accessed, immersed in Lugol for 7days and scanned again. Then, the root canals of 5 teeth were prepared and scanned, and the volume of remaining pulp tissue identified and quantified, whilst 3 teeth were histologically processed. The same specimens were subjected to histological assessment, and the images of the histologic sections were registered with the corresponding micro-CT images to verify whether the pulp tissue in the histological sections matched its counterpart in the Lugol-impregnated tissues identified in the micro-CT slices. There was no discernible effect on radiopacity when NaOCl was mixed with Lugol's solution. Tissue processing did not affect the time required for the complete dissolution of fresh bovine meat. Histological evaluation revealed a correlation between micro-CT and histological images confirming the identification of Lugol-impregnated pulp tissue in micro-CT images. The radiocontrast Lugol's solution was unaffected by NaOCl and did not interfere with its soft tissue dissolution capability. The impregnation protocol using Lugol's solution allowed the visualization of pulp tissue on the micro-CT images and the identification of pulp remnants after chemical-mechanical canal procedures.

Abstract PR02: Profiling intratumoral heterogeneity of bladder cancer subtypes at the single-cell level using machine-learning assisted histopathology

Abstract Purpose: Machine-learning assisted histopathology using markers of basal and luminal differentiation was employed to profile the intratumoral heterogeneity of bladder cancer from cystectomy patients and predict disease-free survival in this high-risk patient population. Methods: Urothelial carcinomas are biologically heterogeneous and vary greatly in clinical progression as well as treatment response. Delineation of molecular subtypes by gene expression analysis of luminal and basal markers has indicated differential outcomes associated with basal and luminal subtypes. However, histologic validation of this classification using protein markers (basal = KRT5/6, P63; luminal = KRT20/GATA3) has been challenging. While using multiplex-immunofluorescence to subtype a retrospective cystectomy cohort (a TMA of 380 patients), we determined that nearly 50% of tumors did not exhibit cytokeratin markers. Subtyping was further confounded by frequent loss of basal-to-luminal stratification and the emergence of intratumoral spatial heterogeneity with the basal and luminal subtypes being completely intermixed throughout the tumor. These observations caused us to hypothesize that previously undefined but clinically relevant subtypes might exist. To address this challenge we developed a single-cell image analysis pipeline that leveraged machine learning to classify molecular subtype and spatial heterogeneity within each tumor. Using the informatics software KNIME we achieved single-cell segmentation and extracted 285 features for 5 protein markers (P63, GATA3, collagen, nuclear stain, and pan-cytokeratin) from each ~20,000 cells contained in 2 cores of tumor and adjacent benign for each patient. Under guidance from a pathologist, definitive urothelial cells (luminal, intermediate, and basal cells) as well as stromal cells were selected from 25 cores normal urothelium to form the ground truth for XGboost-based machine-learning. Summary Findings: Single-cell profiling with machine learning on transcription factors could classify basal and luminal subtypes with greater than 97% accuracy according to validation in normal urothelium using keratin markers. While we were able to recapitulate differential survival associated with a pure basal subtype, it was the intratumoral heterogeneity of basal and luminal cells that was the predominant driver of disease-free survival. Conclusion: A newly identified bladder cancer subtype defined by intratumoral heterogeneity is a clinically relevant driver of disease-free survival. This abstract is also being presented as Poster B22. Citation Format: Adel Eskaros, Tatiana Novitskaya, Andries Zijlstra. Profiling intratumoral heterogeneity of bladder cancer subtypes at the single-cell level using machine-learning assisted histopathology [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2019 May 18-21; Denver, CO. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(15_Suppl):Abstract nr PR02.

DIAGNOSTIC ACCURACY OF APPROXIMAL CARIES IN DIGITAL RADIOGRAPH BY CHINESE AND AMERICAN DENTISTS: AN IN VIVO STUDY

<h3>Background</h3> Because dental education system is not the same in both the United States and China, there may be differences between Chinese and American dentists in the radiographic diagnosis of dental lesions. <h3>Objective</h3> The aim of this study was to assess whether there were any differences in the accuracy of diagnosis of approximal caries between Chinese and American dentists. <h3>Materials and Methods</h3> The study was approved by the local ethical committee. Thirty-nine noncavitated teeth were collected from 11 patients who had parts of the upper or lower jaws excised to remove a cyst or neoplasm. Before surgery, radiographs of the involved teeth were taken by using the digital imaging system Digora Optime (Soredex, Helsinki, Finland). The teeth were then sectioned for histologic validation of the lesions. Four Chinese and 4 American dentists and 5 American senior dental students evaluated all of the radiographs according to a 5-category scale. Receiver operating characteristic (ROC) analysis was performed. Repeated-measure analysis of variance (ANOVA) was employed for statistical analysis. <h3>Results</h3> There were no significant differences among the Chinese and American dentists and the American senior dental students (<i>P</i> = .472). Interobserver (<i>P</i> = .67) and intraobserver (<i>P</i> = .24) variances were not significant. <h3>Discussion</h3> Dental education background is not an influencing factor on the accuracy of diagnosis of approximal caries based on digital radiographs.

Histological validation of a type 1 diabetes clinical diagnostic model for classification of diabetes.

Misclassification of diabetes is common due to an overlap in the clinical features of type 1 and type 2 diabetes. Combined diagnostic models incorporating clinical and biomarker information have recently been developed that can aid classification, but they have not been validated using pancreatic pathology. We evaluated a clinical diagnostic model against histologically defined type 1 diabetes. We classified cases from the Network for Pancreatic Organ donors with Diabetes (nPOD) biobank as type 1 (n=111) or non-type 1 (n=42) diabetes using histopathology. Type 1 diabetes was defined by lobular loss of insulin-containing islets along with multiple insulin-deficient islets. We assessed the discriminative performance of previously described type 1 diabetes diagnostic models, based on clinical features (age at diagnosis, BMI) and biomarker data [autoantibodies, type 1 diabetes genetic risk score (T1D-GRS)], and singular features for identifying type 1 diabetes by the area under the curve of the receiver operator characteristic (AUC-ROC). Diagnostic models validated well against histologically defined type 1 diabetes. The model combining clinical features, islet autoantibodies and T1D-GRS was strongly discriminative of type 1 diabetes, and performed better than clinical features alone (AUC-ROC 0.97 vs. 0.95; P=0.03). Histological classification of type 1 diabetes was concordant with serum C-peptide [median <17pmol/l (limit of detection) vs. 1037pmol/l in non-type 1 diabetes; P<0.0001]. Our study provides robust histological evidence that a clinical diagnostic model, combining clinical features and biomarkers, could improve diabetes classification. Our study also provides reassurance that a C-peptide-based definition of type 1 diabetes is an appropriate surrogate outcome that can be used in large clinical studies where histological definition is impossible. Parts of this study were presented in abstract form at the Network for Pancreatic Organ Donors Conference, Florida, USA, 19-22 February 2019 and Diabetes UK Professional Conference, Liverpool, UK, 6-8 March 2019.

Histological validation of in vivo assessment of cancer tissue inhomogeneity and automated morphological segmentation enabled by Optical Coherence Elastography

We present a non-invasive (albeit contact) method based on Optical Coherence Elastography (OCE) enabling the in vivo segmentation of morphological tissue constituents, in particular, monitoring of morphological alterations during both tumor development and its response to therapies. The method uses compressional OCE to reconstruct tissue stiffness map as the first step. Then the OCE-image is divided into regions, for which the Young’s modulus (stiffness) falls in specific ranges corresponding to the morphological constituents to be discriminated. These stiffness ranges (characteristic "stiffness spectra") are initially determined by careful comparison of the "gold-standard" histological data and the OCE-based stiffness map for the corresponding tissue regions. After such pre-calibration, the results of morphological segmentation of OCE-images demonstrate a striking similarity with the histological results in terms of percentage of the segmented zones. To validate the sensitivity of the OCE-method and demonstrate its high correlation with conventional histological segmentation we present results obtained in vivo on a murine model of breast cancer in comparative experimental study of the efficacy of two antitumor chemotherapeutic drugs with different mechanisms of action. The new technique allowed in vivo monitoring and quantitative segmentation of (1) viable, (2) dystrophic, (3) necrotic tumor cells and (4) edema zones very similar to morphological segmentation of histological images. Numerous applications in other experimental/clinical areas requiring rapid, nearly real-time, quantitative assessment of tissue structure can be foreseen.

Histological validation of adipogenic differentiation potential of ASC on collagen-based 2D scaffolds.

Determination of the adipogenic potential and behavior of adipose-derived mesenchymal stem/stromal cells (ASCs) is particularly relevant for their potential clinical application in regenerative medicine, especially when regeneration is supported by biomaterials or scaffolds. Scaffolds need to be able to induce tissue repair and limit undesired adipogenic differentiation. Depending on the scaffold employed, determination of cell behavior may be hindered by material interference with staining, which will limit either cells identification or dye quantification. Collagen is a promising biomaterial in regenerative medicine, however, histological analysis of cells cultured on collagen-based scaffolds is challenging. Here we describe a new histological method based on iron hematoxylin combined with Oil red O (ORO) staining, for the determination of the adipogenic differentiation of ASCs cultivated on a collagen-based 2D scaffold. ASCs were seeded on collagen films or plastic, differentiated into adipocytes for 14days, and then stained with either ORO or iron hematoxylin and ORO combined. The collagen films avidly absorbed the ORO dye; conventional staining and quantification by dye extraction failed to discriminate between differentiated and undifferentiated cells on the films. On the contrary, the iron hematoxylin-ORO combination provided a quantitative and more reliable determination of adipocytes based on single cell count. This method is particularly recommended for determining the adipogenic differentiation potential of ASCs and other cell types grown on highly absorptive materials that need to be validated for their potential use in bioengineering and regenerative medicine.

The Accumulation of Tau-Immunoreactive Hippocampal Granules and Corpora Amylacea Implicates Reactive Glia in Tau Pathogenesis during Aging.

SummaryThe microtubule-associated tau protein forms pathological inclusions that accumulate in an age-dependent manner in tauopathies including Alzheimer's disease (AD). Since age is the major risk factor for AD, we examined endogenous tau species that evolve during aging in physiological and diseased conditions. In aged mouse brain, we found tau-immunoreactive clusters embedded within structures that are reminiscent of periodic acid-Schiff (PAS) granules. We showed that PAS granules harbor distinct tau species that are more prominent in 3xTg-AD mice. Epitope profiling revealed hypo-phosphorylated rather than hyper-phosphorylated tau commonly observed in tauopathies. High-resolution imaging and 3D reconstruction suggest a link between tau clusters, reactive astrocytes, and microglia, indicating that early tau accumulation may promote neuroinflammation during aging. Using postmortem human brain, we identified tau as a component of corpora amylacea (CA), age-related structures that are functionally analogous to PAS granules. Overall, our study supports neuroimmune dysfunction as a precipitating event in tau pathogenesis.

Prognostic value of myocardial fibrosis in severe aortic stenosis: study protocol for a prospective observational multi-center study (FIB-AS)

BackgroundAdverse cardiac remodeling with a myocardial fibrosis as a key pathophysiologic component may be associated to worse survival in aortic stenosis (AS) patients. Therefore, with the application of advanced cardiac imaging we aim to investigate left ventricular myocardial fibrosis in severe AS patients undergoing aortic valve replacement (AVR) and determine its impact with post-intervention clinical outcomes.MethodsIn a prospective, observational, cohort study patients with severe AS scheduled either for surgical or transcatheter AVR will be recruited from two tertiary heart centers in Denmark and Lithuania. All patients will receive standard of care in accordance with the current guidelines and will undergo additional imaging testing before and after AVR: echocardiography with deformation analysis and cardiovascular magnetic resonance (CMR) with T1 parametric mapping. Those undergoing surgical AVR will also have a myocardial biopsy sampled at the time of a surgery for histological validation. Patients will be recruited over a 2-year period and followed up to 2 years to ascertain clinical outcomes. Follow-up CMR will be performed 12 months following AVR, and echocardiography with deformation analysis will be performed 3, 12, and 24 months following AVR. The study primary outcome is a composite of all-cause mortality and major adverse cardiovascular events.DiscussionDespite continuous effort of research community there is still a lack of early predictors of left ventricular decompensation in AS, which could improve patient risk stratification and guide the optimal timing for aortic valve intervention, before irreversible left ventricular damage occurs. Advanced cardiac imaging and CMR derived markers of diffuse myocardial fibrosis could be utilized for this purpose. FIB-AS study is intended to invasively and non-invasively assess diffuse myocardial fibrosis in AS patients and investigate its prognostic significance in post-interventional outcomes. The results of the study will expand the current knowledge of cardiac remodeling in AS and will bring additional data on myocardial fibrosis and its clinical implications following AVR.Ethics/disseminationThe study has full ethical approval and is actively recruiting patients. The results will be disseminated through scientific journals and conference presentations.Trial registrationClinicalTrials.govNCT03585933. Registered on 02 July 2018.

MR Intracranial Vessel Wall Imaging: A Systematic Review.

The purpose of this systematic review is to identify trends and extent of variability in intracranial vessel wall MR imaging (VWI) techniques and protocols. Although variability in selection of protocol design and pulse sequence type is known, data on what and how protocols vary are unknown. Three databases were searched to identify publications using intracranial VWI. Publications were screened by predetermined inclusion/exclusion criteria. Technical development publications were scored for completeness of reporting using a modified Nature Reporting Summary Guideline to assess reproducibility. From 2,431 articles, 122 met the inclusion criteria. Trends over the last 23 years (1995-2018) show increased use of 3-Tesla MR (P<.001) and 3D volumetric T1-weighted acquisitions (P<.001). Most (65%) clinical VWI publications report achieving a noninterpolated in-plane spatial resolution of ≤.55 mm. In the last decade, an increasing number of technical development (n = 20) and 7 Tesla (n = 12) publications have been published, focused on pulse sequence development, improving cerebrospinal fluid suppression, scan efficiency, and imaging ex vivo specimen for histologic validation. Mean Reporting Summary Score for the technical development publications was high (.87, range: .63-1.0) indicating strong scientific technical reproducibility. Innovative work continues to emerge to address implementation challenges. Gradual adoption into the research and scientific community was suggested by a shift in the name in the literature from "high-resolution MR" to "vessel wall imaging," specifying diagnostic intent. Insight into current practices and identifying the extent of technical variability in the literature will help to direct future clinical and technical efforts to address needs for implementation.

Lesion Size- and Location-Dependent Recruitment of Contralesional Thalamus and Motor Cortex Facilitates Recovery after Stroke in Mice

Brain lesions caused by cerebral ischemia or hemorrhage lead to a local breakdown of energy homeostasis followed by irreversible cell death and long-term impairment. Importantly, local brain lesions also generate remote functional and structural disturbances, which contribute to the behavioral deficit but also impact the recovery of function. While spontaneous recovery has been associated with endogenous repair mechanisms at the vascular, neural, and immune cell levels, the impact of structural plasticity on sensory-motor dysfunction and recovery thereof remains to be elucidated by longitudinal imaging in a mouse model. Here, we applied behavioral assessments, in vivo fiber tracking, and histological validation in a photothrombotic stroke mouse model. Atlas-based whole-brain structural connectivity analysis and ex vivo histology revealed secondary neurodegeneration in the ipsilesional brain areas, mostly in the dorsal sensorimotor area of the thalamus. Furthermore, we describe for the first time a lesion size-dependent increase in structural connectivity between the contralesional primary motor cortex and thalamus with the ipsilesional cortex. The involvement of the contralesional hemisphere was associated with improved functional recovery relative to lesion size. This study highlights the importance of in vivo fiber tracking and the role of the contralesional hemisphere during spontaneous functional improvement as a potential novel stroke biomarker and therapeutic targets.

A Multimodal Ca(II) Responsive Near IR-MR Contrast Agent Exhibiting High Cellular Uptake.

Ca(II) ions are critical for the proper function of neurons by contributing to synaptic signaling and regulating neuronal plasticity. Dysregulation of Ca(II) is associated with a number of pathologies that cause neurodegeneration; therefore the ability to monitor Ca(II) intracellularly is an important target for molecular imaging. Contrast-enhanced MR imaging is a promising modality for imaging changes in Ca(II) concentrations.However, the majority of Ca(II) responsive MR agents are limited to the extracellular space or hindered by poor cellular uptake. Here, we describe a new class of multimodal, bioresponsive Ca(II) magnetic resonance agents that are coupled to the NIR probe IR-783. This new design is based on previous generations of our Ca(II) MR agents but overcomes two significant challenges: (1) the presence of the NIR probe dramatically increases cellular uptake of the agent and (2) provides histological validation of the MR signal using NIR fluorescence imaging. IR-783 targets organic anion transporter polypeptides, and we demonstrate that the agents are not toxic in HT-22 or U-87 MG cells up to 20 μM. The cellular uptake of complex 1 was measured to be greater than 16 femtomoles per cell (where ∼1 femtomole/cell is detectable in acquired MR images). Complex 1 is simultaneously detectable by both MR and NIR fluorescence imaging in vitro and is activated (turned on) by intracellular Ca(II) at concentrations between 1 and 10 μM.

Histological Validation of Cardiovascular Magnetic Resonance T1 Mapping for Assessing the Evolution of Myocardial Injury in Myocardial Infarction: An Experimental Study.

ObjectiveTo determine whether T1 mapping could monitor the dynamic changes of injury in myocardial infarction (MI) and be histologically validated.Materials and MethodsIn 22 pigs, MI was induced by ligating the left anterior descending artery and they underwent serial cardiovascular magnetic resonance examinations with modified Look-Locker inversion T1 mapping and extracellular volume (ECV) computation in acute (within 24 hours, n = 22), subacute (7 days, n = 13), and chronic (3 months, n = 7) phases of MI. Masson's trichrome staining was performed for histological ECV calculation. Myocardial native T1 and ECV were obtained by region of interest measurement in infarcted, peri-infarct, and remote myocardium.ResultsNative T1 and ECV in peri-infarct myocardium differed from remote myocardium in acute (1181 ± 62 ms vs. 1113 ± 64 ms, p = 0.002; 24 ± 4% vs. 19 ± 4%, p = 0.031) and subacute phases (1264 ± 41 ms vs. 1171 ± 56 ms, p < 0.001; 27 ± 4% vs. 22 ± 2%, p = 0.009) but not in chronic phase (1157 ± 57 ms vs. 1120 ± 54 ms, p = 0.934; 23 ± 2% vs. 20 ± 1%, p = 0.109). From acute to chronic MI, infarcted native T1 peaked in subacute phase (1275 ± 63 ms vs. 1637 ± 123 ms vs. 1471 ± 98 ms, p < 0.001), while ECV progressively increased with time (35 ± 7% vs. 46 ± 6% vs. 52 ± 4%, p < 0.001). Native T1 correlated well with histological findings (R2 = 0.65 to 0.89, all p < 0.001) so did ECV (R2 = 0.73 to 0.94, all p < 0.001).ConclusionT1 mapping allows the quantitative assessment of injury in MI and the noninvasive monitoring of tissue injury evolution, which correlates well with histological findings.

Vulnerable plaque imaging using 18F-sodium fluoride positron emission tomography.

Positron emission tomography (PET) with 18F-sodium fluoride (18F-NaF) has emerged as a promising non-invasive imaging modality to identify high-risk and ruptured atherosclerotic plaques. By visualizing microcalcification, 18F-NaF PET holds clinical promise in refining how we evaluate coronary artery disease, shifting our focus from assessing disease burden to atherosclerosis activity. In this review, we provide an overview of studies that have utilized 18F-NaF PET for imaging atherosclerosis. We discuss the associations between traditional coronary artery disease measures (risk factors) and 18F-NaF plaque activity. We also present the data on the histological validation as well as show how 18F-NaF uptake is associated with plaque morphology on intravascular and CT imaging. Finally, we discuss the technical challenges associated with 18F-NaF coronary PET highlighting recent advances in this area.

R2 prime (R2') magnetic resonance imaging for post-myocardial infarction intramyocardial haemorrhage quantification.

To assess whether R2* is more accurate than T2* for the detection of intramyocardial haemorrhage (IMH) and to evaluate whether T2' (or R2') is less affected by oedema than T2* (R2*), and thus more suitable for the accurate identification of post-myocardial infarction (MI) IMH. Reperfused anterior MI was performed in 20 pigs, which were sacrificed at 120 min, 24 h, 4 days, and 7 days. At each time point, cardiac magnetic resonance (CMR) T2- and T2*-mapping scans were recorded, and myocardial tissue samples were collected to quantify IMH and myocardial water content. After normalization by the number of red blood cells in remote tissue, histological IMH increased 5.2-fold, 10.7-fold, and 4.1-fold at Days 1, 4, and 7, respectively. The presence of IMH was correlated more strongly with R2* (r = 0.69; P = 0.013) than with T2* (r = -0.50; P = 0.085). The correlation with IMH was even stronger for R2' (r = 0.72; P = 0.008). For myocardial oedema, the correlation was stronger for R2* (r = -0.63; P = 0.029) than for R2' (r = -0.50; P = 0.100). Multivariate linear regressions confirmed that R2* values were significantly explained by both IMH and oedema, whereas R2' values were mostly explained by histological IMH (P = 0.024) and were little influenced by myocardial oedema (P = 0.262). Using CMR mapping with histological validation in a pig model of reperfused MI, R2'more accurately detected IMH and was less influenced by oedema than R2* (and T2*). Further studies are needed to elucidate whether R2' is also better suited for the characterization of post-MI IMH in the clinical setting.

Efficacy and Reproducibility of Attenuation-Compensated Optical Coherence Tomography for Assessing External Elastic Membrane Border and Plaque Composition in Native and Stented Segments - An In Vivo and Histology-Based Study.

Attenuation-compensated (AC) technique was recently introduced to improve the plaque characterization of optical coherence tomography (OCT). Histological validation demonstrated promising results but the efficacy and reproducibility of this technique for assessing in-vivo tissue composition remains unclear.Methods and Results:OCT images portraying native (n=200) and stented (n=200) segments and 31 histological cross-sections were analyzed. AC-OCT appeared superior to conventional (C)-OCT in detecting the external elastic lamina (EEM) borders (76% vs. 65.5%); AC-OCT enabled larger EEM arc detection compared with C-OCT (174.2±58.7° vs. 137.5±57.9°; P<0.001). There was poor agreement between the 2 techniques for detection of lipid in native and lipid and calcific tissue in stented segments (κ range: 0.164-0.466) but the agreement of C-OCT and AC-OCT was high for calcific tissue in native segments (κ=0.825). Intra and interobserver agreement of the 2 analysts was moderate to excellent with C-OCT (κ range: 0.681-0.979) and AC-OCT (κ range: 0.733-0.892) for all tissue types in both native and stented segments. Ex-vivoanalysis demonstrated that C-OCT was superior to AC-OCT (κ=0.545 vs. κ=0.296) for the detection of the lipid component in native segments. The AC technique allows better delineation of the EEM but it remains inferior for lipid pool detection and neointima characterization. Combined AC- and C-OCT imaging may provide additional value for complete assessment of plaque and neointima characteristics.

Patients Resistant Against PSMA-Targeting α-Radiation Therapy Often Harbor Mutations in DNA Damage-Repair-Associated Genes.

Prostate-specific membrane antigen (PSMA)-targeting α-radiation therapy (TAT) is an emerging treatment modality for metastatic castration-resistant prostate cancer. There is a subgroup of patients with poor response despite sufficient expression of PSMA in their tumors. The aim of this work was to characterize PSMA-TAT-nonresponding lesions by targeted next-generation sequencing. Methods: Of 60 patients treated with 225Ac-PSMA-617, we identified 10 patients who presented with a poor response despite sufficient tumor uptake in PSMA PET/CT. We were able to perform CT-guided biopsies with histologic validation of the nonresponding lesions in 7 of these nonresponding patients. Specimens were analyzed by targeted next-generation sequencing interrogating 37 DNA damage-repair-associated genes. Results: In the 7 tumor samples analyzed, we found a total of 15 whole-gene deletions, deleterious or presumably deleterious mutations affecting TP53 (n = 3), CHEK2 (n = 2), ATM (n = 2), and BRCA1, BRCA2, PALB2, MSH2, MSH6, NBN, FANCB, and PMS1 (n = 1 each). The average number of deleterious or presumably deleterious mutations was 2.2 (range, 0-6) per patient. In addition, several variants of unknown significance in ATM, BRCA1, MSH2, SLX4, ERCC, and various FANC genes were detected. Conclusion: Patients with resistance to PSMA-TAT despite PSMA positivity frequently harbor mutations in DNA damage-repair and checkpoint genes. Although the causal role of these alterations in the patient outcome remains to be determined, our findings encourage future studies combining PSMA-TAT and DNA damage-repair-targeting agents such as poly(ADP-ribose)-polymerase inhibitors.

107Diagnostic accuracy of optical coherence tomography for the identification of in-stent fibroatheroma following stent implantation: an ex-vivo histological validation study

Abstract Background Previous histopathological studies have demonstrated that new atherosclerotic formation within the neointima, called neoatherosclerosis, is one of the most important mechanisms leading to both very late in-stent restenosis and stent thrombosis after stent implantation. Therefore, to distinguish lipid-containing atherosclerotic neointima from other tissues using intracoronary imaging modalities is clinically important to prevent late stent failures. Purpose This study evaluated the diagnostic performance of optical coherence tomography (OCT) for the detection of “in-stent fibroatheroma” following stent implantation by comparing cross-sections of the model with the corresponding histological images. Methods Fifty stented coronary arteries from the 31 autopsy hearts were imaged by OCT. Coronary arterial histopathological specimens, all of which included more than 30% of %neointimal hyperplasia, were compared with the corresponding OCT cross-sections. Histological in-stent fibroatheroma was defined as neointima containing large necrotic core and inflammatory cells. OCT-derived in-stent fibroatheroma comprised a low-intensity tissue containing a poorly delineated region with invisible stent strut behind low signal intensity. Results A total of 122 OCT cross-sections were compared with histological images. OCT examination revealed that 24 images (20%) contained low-intensity tissue inside the neointima. Of those, 5 images, in which stent strut behind low signal intensity was invisible, were diagnosed as OCT-derived in-stent fibroatheroma (4%) (Figure A). By histological analysis, only 4 images were classified as in-stent fibroatheroma (3%) (Figure B). With histology as the gold standard, the sensitivity, specificity, positive predictive value, negative predictive value, and overall diagnostic accuracy for OCT-derived in-stent fibroatheroma were 100%, 99%, 80%, 100%, and 99%, respectively. The only histological finding underlying the false-positive-diagnosis of OCT-derived in-stent fibroatheroma was foam cells accumulation without necrotic core on the neointimal surface (Figure C and D). Most tissue that showed low-intensity tissue with visible stent strut by OCT contained proteoglycan matrix and organized thrombus in the absence of an underlying necrotic core. Coregistration of OCT with histology Conclusion This study showed the potential capability of OCT based on the visualization of stent struts behind low-intensity regions for discriminating in-stent fibroatheroma from other neointimal tissues following stent implantation.