Diagnosis of neonatal neuroblastoma with postmortem magnetic resonance imaging

Postmortem magnetic resonance imaging (MRI) is emerging as a valuable tool to accompany traditional autopsy and has potential for use in cases when traditional autopsy is not possible. This case report will review the use of postmortem MRI with limited tissue sampling to differentiate between metastatic neuroblastoma and hepatoblastoma which could not be clearly differentiated with prenatal ultrasound, prenatal MRI, or emergent postnatal ultrasound. The mother presented to our institution at 27 weeks gestation after an obstetric ultrasound at her obstetrician's office identified a large abdominal mass. Fetal ultrasonography and MRI confirmed the mass but were unable to differentiate between neuroblastoma and multifocal hepatoblastoma. The baby was delivered by cesarean section after nonreassuring heart tones led to an emergent cesarean section. The baby underwent decompressive laparotomy to relieve an abdominal compartment syndrome; however, the family eventually decided to withdraw life support. At this time, we performed a whole body postmortem MRI which further characterized the mass as an adrenal neuroblastoma which was confirmed with limited tissue sampling. Postmortem MRI was especially helpful in this case, as the patient’s family declined traditional autopsy

Metabolomic Profiling of Cerebral Palsy Brain Tissue Reveals Novel Central Biomarkers and Biochemical Pathways Associated with the Disease: A Pilot Study

Cerebral palsy (CP) is one of the most common causes of motor disability in childhood, with complex and heterogeneous etiopathophysiology and clinical presentation. Understanding the metabolic processes associated with the disease may aid in the discovery of preventive measures and therapy. Tissue samples (caudate nucleus) were obtained from post-mortem CP cases (n = 9) and age- and gender-matched control subjects (n = 11). We employed a targeted metabolomics approach using both 1H NMR and direct injection liquid chromatography-tandem mass spectrometry (DI/LC-MS/MS). We accurately identified and quantified 55 metabolites using 1H NMR and 186 using DI/LC-MS/MS. Among the 222 detected metabolites, 27 showed significant concentration changes between CP cases and controls. Glycerophospholipids and urea were the most commonly selected metabolites used to develop predictive models capable of discriminating between CP and controls. Metabolomics enrichment analysis identified folate, propanoate, and androgen/estrogen metabolism as the top three significantly perturbed pathways. We report for the first time the metabolomic profiling of post-mortem brain tissue from patients who died from cerebral palsy. These findings could help to further investigate the complex etiopathophysiology of CP while identifying predictive, central biomarkers of CP

Genome-Wide DNA Methylation Analysis and Epigenetic Variations Associated with Congenital Aortic Valve Stenosis (AVS)

<div><p>Congenital heart defect (CHD) is the most common cause of death from congenital anomaly. Among several candidate epigenetic mechanisms, DNA methylation may play an important role in the etiology of CHDs. We conducted a genome-wide DNA methylation analysis using an Illumina Infinium 450k human methylation assay in a cohort of 24 newborns who had aortic valve stenosis (AVS), with gestational-age matched controls. The study identified significantly-altered CpG methylation at 59 sites in 52 genes in AVS subjects as compared to controls (either hypermethylated or demethylated). Gene Ontology analysis identified biological processes and functions for these genes including positive regulation of receptor-mediated endocytosis. Consistent with prior clinical data, the molecular function categories as determined using DAVID identified low-density lipoprotein receptor binding, lipoprotein receptor binding and identical protein binding to be over-represented in the AVS group. A significant epigenetic change in the <i>APOA5</i> and <i>PCSK9</i> genes known to be involved in AVS was also observed. A large number CpG methylation sites individually demonstrated good to excellent diagnostic accuracy for the prediction of AVS status, thus raising possibility of molecular screening markers for this disorder. Using epigenetic analysis we were able to identify genes significantly involved in the pathogenesis of AVS.</p></div

Receiver operating characteristic (ROC) curve analysis of methylation profiles for four specific markers associated with aortic valve stenosis.

<p>AUC: Area Under Curve; 95% CI: 95% Confidence Interval. Lower and upper confidence interval was given in parentheses. We have identified six CpG sites (cg16464924, cg05890887, cg26196700, cg22469274 cg00994804 cg04836786) among 52 differentially methylated genes that have ROC AUC ≥0.75. At each locus, the FDR p-value for methylation difference between AVS subjects and controls was highly significantly different. Due to figure resolution, we have included only for four markers.</p

Highly Differentially Methylated CpG sites in Aortic Stenosis subjects.

<p>Differentially methylated genes with Target ID, Gene ID, chromosome location, % methylation change and FDR p-value for each gene methylated. CpG sites with significant FDR p-value indicating methylation status and ROC AUC ≥0.75 appear to have a strong potential as diagnostic biomarkers for AVS.</p

Heatmap of unsupervised hierarchical clustering of AVS data on the basis of 59 differentially methylated CpG sites.

<p>Unsupervised hierarchical clustering analysis is very popular in identifying methylation-defined patient sub-groups. Fig 1 displays CpG sites that are at least either 1.5 fold demethylated or 1.5 fold hypermethylated in the disease (AVS) condition (red colored squares) compared to normal subjects (blue colored squares). Differentially methylated CpG sites have been displayed in three clusters (row-wise). The figure also displays direction, fold change in disease, probe relationship and probe annotation of differentially methylated CpG sites. Red color in the heatmap indicates hyper DNA-methylation, and blue hypo DNA-methylation values.</p