Endomyocardial Biopsy Specimens Research Articles

Prevalence of cardiotropic viruses in adults with clinically suspected myocarditis in South Africa

ObjectivesTo determine the prevalence and types of viral pathogens in the myocardium of patients presenting with clinically suspected myocarditis in South Africa.MethodThis is a prospective cross-sectional study. Consecutive adults presenting...

Correlations between pathological deposition and non-invasive diagnostic modalities like 99mTc-PYP scintigraphy, cardiac magnetic resonance, GLS in patients with transthyretin cardiac amyloidosis

Abstract Background Although wild-type transthyretin amyloid cardiomyopathy (ATTRwt-CM) was previously considered a rare disease, recent diagnostic imaging modalities have revealed that it is considerably underdiagnosed among elderly patients with heart failure. The severity of CM is thought to be related to the extent of amyloid deposition in heart. 99mTc-labeled pyrophosphate (99mTc-PYP) scintigraphy, cardiovascular magnetic resonance (CMR), global longitudinal strain (GLS) provide diagnostic and prognostic information in ATTRwt-CM. However, the relevance of these imaging modalities and their association with cardiac amyloid load has not been fully evaluated. Purpose The aim of study was to elucidate the associations between pathological amyloid load and cardiac retention evaluated by 99mTc-PYP scintigraphy, CMR, GLS in patients with ATTRwt-CM. Method Cardiac amyloid load was calculated as (amyloid deposition area/ total myocardium area)×100 using endomyocardial biopsy specimen. Cardiac retention was quantified by heart to contralateral (H/CL) ratio by 99mTc-PYP scintigraphy. Native T1 and extracellular volume (ECV) were obtained by CMR. GLS was analyzed using the 2D echo at the time of diagnosis. Result The mean cardiac amyloid load was 23.0±15.2% (n=57) and correlation with H/CL ratio (1.94±0.36 n=57), native T1 (1426.7±52.5 n=57), ECV (57.9±12.9 n=54), GLS (−9.1±2.4 n=57) were positive (r=0.375 p=0.004, r=0.496 r=0.304 p<0.001, r=0.304 p=0.025, r=0.473 p<0.001). Conclusion Increased cardiac amyloid load correlated with an increased 99mTc-PYP positivity, native T1, ECV, and an impaired GLS. These results suggest that imaging parameters may reflect histological and functional changes due to amyloid deposition in the myocardium. Funding Acknowledgement Type of funding sources: None.

Establishment of Methods Indentifying Genes Associated with Acute Cardiac Cellular Rejection Using a Small Thin Slice Specimen

Purpose Endomyocardial biopsy (EMB) is a gold standard to detect acute cellar cardiac rejection (ACR), but due to its invasiveness, a new non-invasive approach is warranted. Our final goal is to identify new biomarkers corresponding to ACR severity. Using RNA-seq analysis, we found specific gene expression profiles in EMB specimens corresponding to the grades of ISHLT criteria. We also established a transcriptome analysis method utilizing an FFPE (formalin fixed paraffin embedded) EMB section remained after pathological examination. Methods 1. Gene expression profiling in fresh frozen EMB section: 7 cardiac recipients who had pathological ACR were enrolled. They were divided into two groups: patients with before and at rejection ISHLT grade 1A (G-going; N=3) and with at and after treating rejection grade 1A (G-resolving; N=4). Transcriptome analysis was performed using a fresh frozen section (0.5 - 3 mm wide, 7 µm thickness). 2. Establishment of a 3’mRNA-seq method utilizing thin FFPE EMB section: 28 myocardial specimens (grade 0 in 7, 1A in 12, 2 in 2, 3A in 4 and quilty in 3) were analyzed. 3’mRNA-seq libraries were prepared using total RNA isolated from an FFPE section (1-2 mm wide, 5 µm) remained after pathological examination. 3. GSEA (Gene Set Enrichment Analysis) and DGE analysis: After RNA-sequencing, gene expression profiling was analyzed by GSEA and clustered by k-means. Results In G-going, we identified up-regulation of the genes mainly related to ACR, interferon gamma, inflammation, and ILG-JAK-STAT3 signaling, while in G-resolving, we identified down-regulation of these genes. Inflammatory responses were also observed in 3’mRNA-seq analysis using FFPE section. We also separated 270 up-regulating genes in grade3A and 229 ACR related genes associated with ISHLT criteria. In our analysis, most of these identified genes were not listed in AlloMap ®. Conclusion Although these analyses were preliminary because of the 1st analysis stage, these technique for analyzing gene expressions using a tiny myocardial sample may have a role to find new biomarkers associated with ACR.

Diagnostic value of miRNA-101 and miRNA-27 in acute heart transplant rejection

Objective : to determine the diagnostic value of miRNA-101 and miRNA-27 expression levels for acute heart transplant rejection. Materials and methods . The study enrolled 46 heart recipients, among whom were 36 men (78.3%); the average age of recipients was 47.7 = 10.8 (16 to 67) years. Serum microRNA expression levels were measured via quantitative polymerase chain reaction (PCR). Graft rejection was verified through morphological analysis of endomyocardial biopsy specimens. Results. The expression levels of miRNA-101 and miRNA-27 in recipients with acute graft rejection are significantly lower than in recipients without rejection (p = 0.04 and p = 0.03, respectively). When the miRNA-101 expression level is below the determined threshold value, the risk of developing acute graft rejection increases 1.8 times (RR = 1.8 [95% CI 1.13–3.01]). When the miRNA-27 expression level is below the determined threshold value, the risk of developing acute graft rejection increases 1.9 times (RR = 1.9 [95% CI 1.12–3.37]). Simultaneous decrease in the expression levels of miRNA-101 and miRNA-27 below the determined threshold values increases the likelihood of acute graft rejection by 2.0 times (RR = 2.0 [95% CI 1.16–3.36]). Conclusion . The serum miRNA-101 and miRNA-27 expression levels are of diagnostic value for acute graft rejection in heart recipients.

The Association between the Clinical Severity of Heart Failure and Docosahexaenoic Acid Accumulation in Hypertrophic Cardiomyopathy

Background: Hypertrophic cardiomyopathy (HCM) is a common genetic disease with diverse morphology, symptoms, and prognosis. Hypertrophied myocardium metabolism has not been explored in detail. We assessed the association between myocardium lipid metabolism and clinical severity of heart failure (HF) in HCM using imaging mass spectrometry (IMS). Methods and Results: We studied 16 endomyocardial biopsy (EMB) specimens from patients with HCM. Analysis was conducted using desorption electrospray ionization IMS. The Samples were assigned into two cohorts according to the period of heart biopsy (cohort 1, n=9 and cohort 2, n=7). In each cohort, samples were divided into two groups according to the clinical severity of HF in HCM: clinically severe and clinically mild groups. Signals showing a significant difference between the two groups were analyzed by volcano plot. In cohort 1, the volcano plot identified four signals; the intensity in the clinically severe group was more than twice that of the mild group. Out of the four signals, docosahexaenoic acid (DHA) showed significant differences in intensity between the two groups in cohort 2 (10575.8±2750.3 vs. 19839.3±4803.2, p=0.025). Conclusion: The intensity of DHA was significantly higher in EMB samples from the clinically severe HCM group than in those from the mild group. Funding Information: This study received funding from the Ministry of Education, Science, Sports and Culture of Japan, Grants-in-Aid for Scientific Research (Grant Number 18K08065 [YM]). Declaration of Interests: The authors have no conflicts of interest to disclose. Ethics Approval Statement: All the patients provided informed consent. The institutional review board of Hamamatsu University School of Medicine approved all aspects of this study (application number 17-261). The study was conducted in accordance with the regulations of Declaration of Helsinki.

Active Cardiac Sarcoidosis Imitating Cardiac Metastases.

A 54-year-old woman diagnosed with sarcoidosis based on mediastinal lymph node biopsy underwent conventional contrast material–enhanced chest CT examination without electrocardiographic synchronization to evaluate pulmonary sarcoidosis. Past medical history included prior left total mastectomy for breast cancer (Fig 1, A). CT results demonstrated multiple hypoattenuating masses involving the septum, free walls of both ventricles, and right atrium (Fig 1, B, C). Cardiac metastases were a potential diagnosis; however, the cardiac lesions were similar in appearance to lymph nodes noted in the hilum and mediastinum. Subsequent late gadolinium enhancement MRI results showed abnormal foci of delayed enhancement in both ventricles and the right atrium, corresponding to the lesions on chest CT (Fig 1, D). Fluorine 18 fluorodeoxyglucose PET results showed abnormal uptake in these areas (maximum standardized uptake value, 12.0) (Fig 1, E). Finally, results of endomyocardial biopsy from the right ventricle revealed a granuloma with lymphocytes, demonstrating active sarcoidosis (Fig 1, F). Open in a separate window A, Contrast-enhanced CT image obtained with a delay time of 90 seconds after contrast medium injection at a slice thickness of 5 mm shows hilar and mediastinal lymphadenopathy (arrows). Note the prior left total mastectomy (arrowheads). B and C, Contrast-enhanced CT images demonstrate hypoattenuating foci in the septum, both ventricles, and right atrium (arrows). D, A four-chamber view of late enhancement MRI acquired 10 minutes after injection of gadolinium-based contrast material reveals delayed gadolinium enhancement in the septum, both ventricles, and right atrium (arrows). E, Abnormal fluorine 18 fluorodeoxyglucose (18F-FDG) uptake is observed in the septum, both ventricles, and right atrium on the 18F-FDG PET image (arrows). F, Endomyocardial biopsy specimen obtained from the right ventricle demonstrates granulomas with giant cells. (CD68 immunohistochemical staining; magnification rate, ×400.)

Diagnostic value of cardiovascular magnetic resonance in comparison to endomyocardial biopsy in cardiac amyloidosis: a multi-centre study

BackgroundCardiac amyloidosis (CA) is an infiltrative disease characterised by accumulation of amyloid deposits in the extracellular space of the myocardium—comprising transthyretin (ATTR) and light chain (AL) amyloidosis as the most frequent subtypes. Histopathological proof of amyloid deposits by endomyocardial biopsy (EMB) is the gold standard for diagnosis of CA. Cardiovascular magnetic resonance (CMR) allows non-invasive workup of suspected CA. We conducted a multi-centre study to assess the diagnostic value of CMR in comparison to EMB for the diagnosis of CA.MethodsWe studied N = 160 patients characterised by symptoms of heart failure and presence of left ventricular (LV) hypertrophy of unknown origin who presented to specialised cardiomyopathy centres in Germany and underwent further diagnostic workup by both CMR and EMB. If CA was diagnosed, additional subtyping based on EMB specimens and monoclonal protein studies in serum was performed. The CMR protocol comprised cine- and late-gadolinium-enhancement (LGE)-imaging as well as native and post-contrast T1-mapping (in a subgroup)—allowing to measure extracellular volume fraction (ECV) of the myocardium.ResultsAn EMB-based diagnosis of CA was made in N = 120 patients (CA group) whereas N = 40 patients demonstrated other diagnoses (CONTROL group). In the CA group, N = 114 (95%) patients showed a characteristic pattern of LGE indicative of CA. In the CONTROL group, only 1/40 (2%) patient showed a “false-positive” LGE pattern suggestive of CA. In the CA group, there was no patient with elevated T1-/ECV-values without a characteristic pattern of LGE indicative of CA. LGE-CMR showed a sensitivity of 95% and a specificity of 98% for the diagnosis of CA. The combination of a characteristic LGE pattern indicating CA with unremarkable monoclonal protein studies resulted in the diagnosis of ATTR-CA (confirmed by EMB) with a specificity of 98% [95%-confidence interval (CI) 92–100%] and a positive predictive value (PPV) of 99% (95%-CI 92–100%), respectively. The EMB-associated risk of complications was 3.13% in this study—without any detrimental or persistent complications.ConclusionNon-invasive CMR shows an excellent diagnostic accuracy and yield regarding CA. When combined with monoclonal protein studies, CMR can differentiate ATTR from AL with high accuracy and predictive value. However, invasive EMB remains a safe invasive gold-standard and allows to differentiate CA from other cardiomyopathies that can also cause LV hypertrophy.

Prognostication of cardiac amyloidosis by pittsburgh b compound positron emission tomography

Abstract Introduction Regardless of the underlying pathogenesis of amyloid production, cardiac involvement is the leading cause of morbidity and mortality in systemic amyloidosis. In the cardiac variant, there is an extracellular deposition of amyloid substance throughout the heart. The infiltrative process results in ventricular wall thickening with concentric hypertrophy and low cardiac output. The PiB compound has been promising for the detection of amyloid deposits in the brain but it remains unknown whether this noninvasive evaluation can predict the prognosis of patients with cardiac amyloidosis. Purpose This study sought to investigate the efficacy of 11C-Pittsburgh B (PiB) positron emission tomography for prognostication of cardiac amyloidosis by noninvasively imaging the myocardial amyloid deposition. Methods A total of 33 consecutive patients were enrolled in this prospective pilot study with suspected cardiac amyloidosis. The amyloid deposit was assessed with amyloid P immunohistochemistry in endomyocardial biopsy specimens and was compared with the degree of myocardial 11C-PiB uptake on PET. Results It was found that the degree of myocardial 11C-PiB PET uptake was significantly greater in the patients of cardiac amyloidosis compared with control group subjects and was parallel with the degree of amyloid deposit on histology (p<0.001). When the cardiac amyloidosis patients were divided into tertiles by the degree of myocardial 11C-PiB PET uptake, patients with the highest PiB uptake experienced the worst clinical event-free survival (log-rank p=0.013). The clinical outcome was significantly predicted by the degree of myocardial PiB PET uptake on multivariate Cox regression analysis (adjusted hazard ratio: 1.191; 95% confidence interval: 1.062 to 1.340; p=0.005). Conclusion Our study concludes that the noninvasive evaluation of myocardial amyloid load by 11C-PiB PET reflects the degree of amyloid deposit and is an independent predictor of clinical outcome in cardiac amyloidosis patients. 11-C Pittsburgh B PET imaging Funding Acknowledgement Type of funding source: Other. Main funding source(s): Self funding

MicroRNA expression levels in early and long-term period following heart transplantation

Objective : to conduct comparative analysis of the expression levels of microRNA-101, microRNA-142, microRNA- 27, microRNA-339 and microRNA-424 in patients with severe chronic heart failure and in heart recipients in the early and long-term period following heart transplantation and to determine the association with acute transplant rejection. Materials and methods . The study included 46 heart recipients, among whom were 36 men (78.3%); the average age of the recipients was 47.7 ± 10.8 (16 to 67) years, and 12 patients with end-stage chronic heart failure, among whom were 8 men (66.7%); the average age of the patients was 46.1 ± 6.4 (37 to 64) years. The control group consisted of 12 healthy individuals, not significantly different by gender and age. microRNA expression levels in blood plasma were determined through quantitative polymerase chain reaction (Q-PCR). Transplant rejection was verified via morphological analysis of endomyocardial biopsy specimens. Results . Blood plasma of patients with end-stage chronic heart failure had significantly higher expression rates of microRNA-101, microRNA-27, microRNA-339 and microRNA-424 than in healthy individuals (p < 0.02). In the early stages following transplantation, the expression levels of microRNA-101 and microRNA-27 in heart recipients were significantly lower than in patients with severe chronic heart failure (p < 0.003). A year or more after transplantation, there were no significant differences in the expression levels of microRNA-101, microRNA- 142, and microRNA-339 in heart recipients and in healthy individuals. In recipients with acute rejection, the expression levels of microRNA-101 and microRNA-27 significantly differed from that of recipients without signs of rejection (p = 0.04 and p = 0.03, respectively). Conclusion . The obtained data on changes in the expression levels of microRNA-101 and microRNA-27 in heart recipients with acute transplant rejection suggests possible diagnostic value of these biomarkers in determining the risk of rejection.

Diagnostic value of galectin-3 in heart transplant recipients with myocardial complications

Objective : to determine the diagnostic value of galectin-3 in transplant recipients with myocardial fibrosis and acute heart transplant rejection, verified by endomyocardial biopsy. Materials and methods . The study included 124 patients with end-stage heart failure. Their ages ranged from 16 to 71 (average 48 ± 12) years, of which 106 (85%) were men and 18 (15%) were women. From 2013 to 2016, these patients underwent a heart transplant procedure at the Shumakov National Medical Research Center of Transplantology and Artificial Organs, Moscow, Russian Federation. Analysis of endomyocardial biopsy specimens was used to verify acute cellular, humoral rejection and myocardial fibrosis of the heart transplant. Severity and nature of fibrosis was evaluated using a qualitative imaging technique. Galectin-3 concentration was measured by enzyme immunoassay using Human Galectin-3 Platinum ELISA reagent kits (Bender MedSystems GmbH, Vienna, Austria). Results . In the long-term post-transplantation period, in comparison with the early post-transplantation period, the number of verified graft myocardial fibrosis increased by 88% in recipients who had acute rejection crises and by 37% in recipients who had no rejection crises. Graft myocardial fibrosis was detected more often in recipients who had acute humoral rejection than in those who had acute cell rejection (92% vs 75% of cases, respectively). Plasma galectin-3 levels in recipients with graft myocardial fibrosis was higher than in recipients without it (p = 0.05 1 year and p = 0.01 1–5 years after heart transplantation). In recipients who had acute rejection crises, the risk of developing graft myocardial fibrosis was 1.64 (RR = 1.64 ± 0.1 [95% CI 1.1–2.2]). Conclusion . Galectin-3 is a biomarker for myocardial fibrosis in acute heart transplant rejection.

Quadritherapy vs standard tritherapy immunosuppressant regimen after heart transplantation: A propensity score-matched cohort analysis.

After heart transplant, adding everolimus (EVL) to standard immunosuppressive regimen mostly relies on converting calcineurin inhibitors (CNIs) into EVL. The aim of this study was to describe the effects of combining low-dose EVL and CNIs in maintenance immunosuppression regimen (quadritherapy) and compare it with standard tritherapy associating standard-dose CNIs, mycophenolate mofetil, and corticosteroids. In the 3-year registry cohort of heart transplanted patients, those who received quadritherapy were compared with those who received tritherapy. EVL was added after 3months posttransplant. Three analyses were performed to control for confounders: propensity score matching, multivariable survival, and inverse probability score weighting analyses. Among 213 patients who were included (75 with quadritherapy), propensity score matching selected 64 unique pairs of patients with similar characteristics. In the matched cohort (n=128), quadritherapy was associated with fewer deaths (3 [4.7%] vs 17 [21.9%], P=.007) and biopsy-proven acute rejections (15 [23.4%] vs 31 [48.4%], P=.002). These results were confirmed in the overall cohort (n=213), after multivariable and inverse probability score weighting analyses. Renal function and donor-specific HLA-antibodies remained similar in both groups. Low-dose combination quadritherapy was associated with fewer deaths and rejections, compared with standard immunosuppression tritherapy.

RNA Expression Analysis of Antibody Mediated Rejection after Heart Transplant with or without Donor Specific Antibodies

Antibody mediated rejection (AMR) is an increasingly studied cause of graft failure after heart transplantation. AMR diagnostic criteria previously required serum donor specific antibodies (DSA). However, the most recent criteria only requires pathological findings independent of DSA. This classification defined a subset of patients with AMR without known antibodies. To investigate this complication, we performed RNA sequencing using endomyocardial biopsy samples taken at the time of pathology confirmed AMR in the presence and absence of DSA. We evaluated the transcriptome profile of the paraffin-embedded endomyocardial biopsy specimens of 30 representative heart transplant recipients. Patients were categorized as having no evidence of AMR (controls), immunohistochemical findings of AMR (pAMR1i) in the presence of DSA (1i+D), pAMR1i in the absence of DSA (1i-D), histopathological findings of AMR (pAMR1h) in the presence of DSA (1h+D), pAMR1h in the absence of DSA (1h-D) or presence of both immunohistochemical and histopathological findings of AMR (pAMR2) in the presence of DSA. A total of 5 patients were analyzed per group. All were matched for previous mechanical circulatory support and time from transplant to biopsy. There was no differential RNA expression between patients with no AMR and those in the 1i-D group. There was also no differential expression between patients in the 1h+D and pAMR2 groups. The largest difference was seen between the no AMR and 1i+D groups and the no AMR and pAMR2 groups. In addition, there was RNA expression differences between patients with immunohistochemical evidence of AMR compared to those with histopathological findings, suggesting distinct mechanisms. These studies demonstrate that RNA expression profiling can be performed on endomyocardial biopsy tissue preserved in paraffin from the time of a rejection episode and that different expression patterns are observed dependent on the criteria used to diagnose AMR. In particular, AMR in the presence of DSA showed a different RNA expression profile than AMR without DSAs. The RNA expression associated with histological markers of AMR also appeared different than that associated with immunological markers. Overall, our findings suggest the existence of different subtypes of AMR that may rely on distinct mechanisms of injury.

Evaluating Biventricular Myocardial Velocity and Interventricular Dyssynchrony in Adult Patients During the First Year After Heart Transplantation.

Magnetic resonance tissue phase mapping (TPM) measures three-directional myocardial velocities of the left and right ventricle (LV, RV). This noninvasive technique may supplement endomyocardial biopsy (EMB) in monitoring grafts post-heart transplantation (HTx). To assess biventricular myocardial velocity alterations in grafts and investigate the relationship between velocities and acute cellular rejection (ACR) episodes. Prospective. Twenty-seven patients within 1 year post-HTx (49 ± 13 years, 19 M) and 18 age-matched controls (49 ± 15 years, 12 M). 1.5T, 2D balanced steady-state free precession, and TPM. Ventricular function: end-diastolic and end-systolic volumes, stroke volumes, ejection fraction (EF), and myocardial mass. TPM velocities: peak-systolic and peak-diastolic velocities, cardiac twist, and interventricular dyssynchrony. ACR rejection episodes: International Society for Heart and Lung Transplantation grading of EMB specimens. The Lilliefors test for normality, unpaired t-tests, and Wilcoxon rank-sum tests for normally and nonnormally distributed data, respectively, were used, as well as multivariate regression for confounding variables and Pearson's correlation for associations between TPM velocities and global function. Compared to controls, HTx patients demonstrated reduced biventricular systolic longitudinal velocities (LV: 5.2 ± 2.1 vs. 4.0 ± 1.5 cm/s, P < 0.05; RV: 4.2 ± 1.3 vs. 3.1 ± 1.2 cm/s, P < 0.01). Correlation analysis revealed significant positive relationships for biventricular EF with radial peak velocities of the same ventricle in both systole and diastole (LV systole: r = 0.48, P < 0.01; LV diastole: r = 0.28, P < 0.05; RV systole: r = 0.35, P < 0.01; RV diastole: r = 0.36, P < 0.01). Segmentally, longitudinal velocities were impaired in 7/16 LV segments and 5/10 RV segments in systole and 7/10 RV segments in diastole. TPM analysis in studies with >4 preceding ACR episodes showed globally reduced RV and LV systolic radial velocity, and segmentally reduced radial and longitudinal systolic velocities. Biventricular global and segmental velocities were reduced in HTx patients. Patients with >4 rejection episodes showed reduced myocardial velocities. The TPM sequence may add functional information for monitoring graft dysfunction. 2 TECHNICAL EFFICACY STAGE: 2 J. Magn. Reson. Imaging 2020;52:920-929.

Pittsburgh B Compound Positron Emission Tomography in Patients With AL Cardiac Amyloidosis

BackgroundIt remains unknown whether the noninvasive evaluation of the degree of amyloid deposition in the myocardium can predict the prognosis of patients with light chain (AL) cardiac amyloidosis. ObjectivesThe purpose of this study was to demonstrate that 11C-Pittsburgh B compound positron emission tomography (11C-PiB PET) is useful for prognostication of AL cardiac amyloidosis by noninvasively imaging the myocardial AL amyloid deposition. MethodsThis study consecutively enrolled 41 chemotherapy-naïve AL cardiac amyloidosis patients. The amyloid deposit was quantitatively assessed with amyloid P immunohistochemistry in endomyocardial biopsy specimens and was compared with the degree of myocardial 11C-PiB uptake on PET. The primary endpoint was a composite of all-cause death, heart transplantation, and acute decompensated heart failure. ResultsThe degree of myocardial 11C-PiB PET uptake was significantly higher in the cardiac amyloidosis patients compared with normal subjects and correlated well with the degree of amyloid deposit on histology (R2 = 0.343, p < 0.001). During follow-up (median: 423 days, interquartile range: 93 to 1,222 days), 24 patients experienced the primary endpoint. When the cardiac amyloidosis patients were divided into tertiles by the degree of myocardial 11C-PiB PET uptake, patients with the highest PiB uptake experienced the worst clinical event-free survival (log-rank p = 0.014). The degree of myocardial PiB PET uptake was a significant predictor of clinical outcome on multivariate Cox regression analysis (adjusted hazard ratio: 1.185; 95% confidence interval: 1.054 to 1.332; p = 0.005). ConclusionsThese proof-of-concept results show that noninvasive evaluation of myocardial amyloid load by 11C-PiB PET reflects the degree of amyloid deposit and is an independent predictor of clinical outcome in AL cardiac amyloidosis patients.

In situ nuclear DNA methylation in dilated cardiomyopathy: an endomyocardial biopsy study.

AimsAlthough distinct DNA methylation patterns have been reported, its localization and roles remain to be defined in heart failure. We investigated the cellular and subcellular localization of DNA methylation and its pathophysiological significance in human failing hearts.Methods and resultsUsing left ventricular (LV) endomyocardial biopsy specimens from 75 patients with dilated cardiomyopathy (DCM; age: 58 ± 14 years old, %female: 32%) and 20 patients without heart failure (controls; age: 56 ± 17 years old, %female: 45%), we performed immunohistochemistry and immunoelectron microscopy for methylated DNA, 5‐methylcytosine (5‐mC). We next investigated possible relations of the incidence of 5‐mC‐positive (%5‐mC+) cardiomyocytes with clinicopathological parameters. Immunopositivity for 5‐mC was detected in the cardiomyocytes and other cell types. The %5‐mC+ cardiomyocytes was significantly greater in DCM hearts than in controls (57 ± 13% in DCM vs. 25 ± 12% in controls, P < 0.0001). The localization of 5‐mC immunopositivity in cardiomyocyte nuclei coincided well with that of heterochromatin, as confirmed by immunoelectron microscopy. Substantial DNA methylation was also observed in interstitial non‐cardiomyocytes, but the incidences did not differ between control and DCM hearts (39 ± 7.9% in DCM vs. 41 ± 10% in controls, P = 0.4099). In DCM patients, the %5‐mC+ cardiomyocytes showed a significant inverse correlation with LV functional parameters such as heart rate (r = 0.2391, P = 0.0388), end‐diastolic pressure (r = 0.2397, P = 0.0397), and ejection fraction (r = −0.2917, P = 0.0111) and a positive correlation with LV dilatation (volume index at diastole; r = 0.2442, P = 0.0347; and volume index at systole; r = 0.3136, P = 0.0062) and LV hypertrophy (mass index; r = 0.2287, P = 0.0484)—that is, LV remodelling parameters. No significant correlations between DNA methylation and the histological parameters of the biopsies, including cardiomyocyte hypertrophy, fibrosis, and inflammatory cell infiltration, were noted.ConclusionsThe present study revealed increased nuclear DNA methylation in cardiomyocytes, but not other cell types, from DCM hearts, with predominant localization in the heterochromatin. Its significant relations with LV functional and remodelling parameters imply a pathophysiological significance of DNA methylation in heart failure.

Supplemental Cardioplegia During Donor Heart Implantation: A Systematic Review and Meta-Analysis

The optimal donor heart preservation and management strategy during heart transplantation remains controversial. We report the results of a systematic review and meta-analysis of the effect of supplemental cardioplegia administration during donor heart implant for transplantation. We searched MEDLINE and Embase databases until February 2019 for studies comparing patients who received transplants with the donor heart given supplemental cardioplegia or not. Data were extracted by 2 independent investigators. The main outcomes were early morbidity and mortality. Included were 7 retrospective observational studies (4 comparing to historical controls) and 3 randomized controlled trials enrolling 1125 patients. Supplemental cardioplegia included crystalloid and blood cardioplegia given continuous retrograde or as terminal "hot shots." Supplemental cardioplegia was associated with improved early mortality (risk ratio [RR], 0.55; 95% confidence interval [CI], 0.35-0.87; P < .01), greater rates of spontaneous return of sinus rhythm (RR, 2.62; 95% CI, 1.50-4.56; P < .01), shorter intensive care stay (mean difference,-3.4 days; 95% CI,-5.1 to-1.6; P < .01), and lower incidence of ischemic changes seen on endomyocardial biopsy specimens (RR, 0.49; 95% CI, 0.35-0.69; P < .01) compared with controls. Midterm mortality was not different between groups (incident rate ratio, 0.80; 95% CI, 0.51-1.26; P= .34). Administration of supplemental cardioplegia may be associated with a reduction in organ ischemic injury and shorter intensive care stay as well as improvement in early survival after transplantation. This strategy may be a simple and cost-effective adjunct to improve outcomes of heart transplantation, especially in an era of increasing use of marginal donor organs. Further investigation will be needed to confirm the findings of this hypothesis-generating study.

Immunoadsorption for Acute Symptomatic Antibody-Mediated Rejection after Heart Transplantation

Purpose Antibody-mediated rejection (AMR) after heart transplantation (HTX) is associated with adverse outcomes . A standardized therapeutic regimen has not yet been defined. Immunoadsorption (IA) has shown promising results in AMR after kidney transplantation . We present our results with IA for acute symptomatic AMR after HTX. Methods This retrospective single-center study included all consecutive patients with proven and treated symptomatic AMR between 2006 and 2016. Survival from AMR, incidence of cardiac allograft vasculopathy (CAV) as well as donor specific anti-HLA-antibody (DSA) cumulative mean fluorescence intensity (MFI) assessed by LUMINEX single antigen bead assay are described. Results 22 patients were designated to undergo extracorporeal antibody removal for symptomatic AMR and received IA (n=18, 81,8 %) or plasmapheresis (PP; n=1, 4,5 %). One patient was switched to IA after one initial PP. In two patients IA/PP was not initiated due to unfavorable prognosis. Concomitant therapy was methylprednisolone (100 %), IVIg (9 %) and rituximab (4,5 %). AMR occurred after a median of 5.65 months (IQR 0.43-21.4) from HTX. Overall survival after a median follow up of 4.85 years from AMR diagnosis was 63.6 %. Mortality 30 days, 1 year and 5 years after AMR onset was 27.3 %, 31.8 % and 31.8 %. 4 patients (18.2 %) developed severe CAV (ISHLT grade ≥ 2) after a median time of 30.5 months after AMR onset. 21 patients had positive DSA at the time of AMR diagnosis (HLA I only: 13.6 %, HLA II only: 50 %, HLA I+II: 31.8%). Conclusion Despite high early mortality, long-term outcome of patients surviving the acute phase is encouraging, however we observed an increased burden of CAV. Due to the oftentimes fulminant clinical course, rapid diagnosis and therapy, facilitated by close interdisciplinary cooperation of intensivists , cardiac surgeons and nephrologists is crucial. Next steps include a detailed characterization of DSAs over the treatment period as well as endomyocardial biopsy specimens of patients with AMR.

Effect of recipient gender and donor-specific antibodies on antibody-mediated rejection after heart transplantation.

Gender-difference regarding antibody-mediated rejection (AMR) after heart transplantation has been described. However, no study accounted for the presence of preformed donor-specific antibodies (pfDSA), a known risk factor of AMR, more common among women than men. In a single-institution 6-year cohort (2010-2015), time to AMR was assessed, comparing men with women by survival analysis with a 1-year death-censored follow-up. All AMRs were biopsy proven. Confounding variables that were accounted for included mean intensity fluorescence (MFI) of pfDSA, recipient age, HLA-, size- and sex-mismatch. 463 patients were included. Overall incidence of AMR was 10.3% at 1year. After adjusting for confounding variables, independent risk factors of AMR were female recipient gender (adjusted hazard-ratio [adj. HR]=1.78 [1.06-2.99]), P=.03) and the presence of pfDSA (adj. HR=3.20 [1.80-5.70], P<.001). This association remained significant when considering pfDSA by their MFI; female recipient gender had an adj. HR=2.2 (P=.026) and MFI of pfDSA (per 1 MFI-increase) adj. HR=1.0002 (P<.0001). In this cohort, women were at higher risk of AMR than men and this risk increase was additive to that of pfDSA. These findings may suggest a gender-related difference in the severity of pfDSA.

Evidence-Based Review and Discussion Points

Evidence-Based Review and Discussion Points

The ratio of circulating regulatory cluster of differentiation 4 T cells to endothelial progenitor cells predicts clinically significant acute rejection after heart transplantation

The aim of this study was to determine the value of the ratio of the percentage of circulating regulatory cluster of differentiation 4 T cells (%Tregs) to the percentage of endothelial progenitor cells (%EPCs; Treg/EPC ratio) for predicting clinically significant acute rejection. Peripheral blood %Tregs and %EPCs were quantified in 91 cardiac transplant recipients using flow cytometry at a mean of 42 ± 13 days after transplant. The primary end point was clinically significant acute rejection, defined as an event that led to an acute augmentation of immunosuppression in conjunction with an International Society for Heart and Lung Transplantation grade ≥ 2R in a right ventricular endomyocardial biopsy specimen or non-cellular rejection (specimen-negative rejection) with hemodynamic compromise (decrease in left ventricular ejection fraction by > 25%). Significant rejection occurred in 27 recipients (29.7%) during a median of 49.4 months (interquartile range, 37.0-62.0 months). The mean %Tregs and %EPCs were not significantly different between those with and without an episode of significant rejection, but the mean Treg/EPC ratio was significantly lower in recipients with significant rejection (44.9 vs 106.7, p = 0.001). Receiver operating characteristic curve analysis showed anarea under the curve value for significant rejection for a Treg/EPC ratio of 0.712. The best cutoff value of the Treg/EPC ratio that distinguished between those with or without significant rejection was ≤ 18 by receiver operating characteristic curve analysis. Kaplan-Meier analysis revealed that patients with a Treg/EPC ratio of ≤ 18 had a significantly higher rate of rejection than those with a Treg/EPC ratio > 18 (61.5% vs 16.9%, log-rank p < 0.0001). A low Treg/EPC ratio was an independent predictor of significant rejection. A low Treg/EPC ratio measured soon after heart transplantation is an independent predictor of acute rejection. The Treg/EPC ratio has potential as an early biomarker after heart transplantation for predicting acute rejection.