Bovine spongiform encephalopathy infection alters endogenous retrovirus expression in distinct brain regions of cynomolgus macaques (Macaca fascicularis)

<p>Abstract</p> <p>Background</p> <p>Prion diseases such as bovine spongiform encephalopathies (BSE) are transmissible neurodegenerative diseases which are presumably caused by an infectious conformational isoform of the cellular prion protein. Previous work has provided evidence that in murine prion disease the endogenous retrovirus (ERV) expression is altered in the brain. To determine if prion-induced changes in ERV expression are a general phenomenon we used a non-human primate model for prion disease.</p> <p>Results</p> <p>Cynomolgus macaques (<it>Macaca fasicularis</it>) were infected intracerebrally with BSE-positive brain stem material from cattle and allowed to develop prion disease. Brain tissue from the <it>basis pontis </it>and <it>vermis cerebelli </it>of the six animals and the same regions from four healthy controls were subjected to ERV expression profiling using a retrovirus-specific microarray and quantitative real-time PCR. We could show that Class I gammaretroviruses HERV-E4-1, ERV-9, and MacERV-4 increase expression in BSE-infected macaques. In a second approach, we analysed ERV-K-(HML-2) RNA and protein expression in extracts from the same cynomolgus macaques. Here we found a significant downregulation of both, the macaque ERV-K-(HML-2) Gag protein and RNA in the frontal/parietal cortex of BSE-infected macaques.</p> <p>Conclusions</p> <p>We provide evidence that dysregulation of ERVs in response to BSE-infection can be detected on both, the RNA and the protein level. To our knowledge, this is the first report on the differential expression of ERV-derived structural proteins in prion disorders. Our findings suggest that endogenous retroviruses may induce or exacerbate the pathological consequences of prion-associated neurodegeneration.</p

Upregulation of miRNA hsa-miR-342-3p in experimental and idiopathic prion disease

The aim of our study was to analyze the differential expression of miRNAs in the brains of BSE-infected cynomolgus macaques as a model for Creutzfeldt-Jakob disease (CJD). MicroRNAs (miRNAs) are small noncoding RNAs regulating gene expression by mRNA targeting. Among other functions they contribute to neuronal development and survival. Recently, the lack of miRNA processing has been shown to promote neurodegeneration and deregulation of several miRNAs has been reported to be associated with Scrapie in mice. Therefore, we hypothesized that miRNAs are also regulated in response to human prion disease. We have applied miRNA-microarrays to identify deregulated miRNA candidates in brains of BSE-infected macaques. Shock-frozen brain sections of six BSE-infected and five non-infected macaques were used to validate regulated miRNA candidates by two independent qRT-PCR-based methods. Our study revealed significant upregulation of hsa-miR-342-3p and hsa-miR-494 in the brains of BSE-infected macaques compared to non-infected animals. In a pilot study we could show that hsa-miR-342-3p was also upregulated in brain samples of human type 1 and type 2 sporadic CJD. With respect to the reported regulation of this miRNA in Scrapie-infected mice, we propose that upregulation of hsa-miR-342-3p may be a general phenomenon in late stage prion disease and might be used as a novel marker for animal and human TSEs

Badinage Woodwind Quintet: Judith Ross, Flute; Wayne Montag, Clarinet; Marvin Carlton, Oboe; Joyce Hitchcock, Bassoon; Richard Weyhrich, Horn; February 10, 1975

Hayden AuditoriumMonday EveningFebruary 10, 19758:15 p.m

Unequal allelic expression of wild-type and mutated β-myosin in familial hypertrophic cardiomyopathy

Familial hypertrophic cardiomyopathy (FHC) is an autosomal dominant disease, which in about 30% of the patients is caused by missense mutations in one allele of the β-myosin heavy chain (β-MHC) gene (MYH7). To address potential molecular mechanisms underlying the family-specific prognosis, we determined the relative expression of mutant versus wild-type MYH7-mRNA. We found a hitherto unknown mutation-dependent unequal expression of mutant to wild-type MYH7-mRNA, which is paralleled by similar unequal expression of β-MHC at the protein level. Relative abundance of mutated versus wild-type MYH7-mRNA was determined by a specific restriction digest approach and by real-time PCR (RT-qPCR). Fourteen samples from M. soleus and myocardium of 12 genotyped and clinically well-characterized FHC patients were analyzed. The fraction of mutated MYH7-mRNA in five patients with mutation R723G averaged to 66 and 68% of total MYH7-mRNA in soleus and myocardium, respectively. For mutations I736T, R719W and V606M, fractions of mutated MYH7-mRNA in M. soleus were 39, 57 and 29%, respectively. For all mutations, unequal abundance was similar at the protein level. Importantly, fractions of mutated transcripts were comparable among siblings, in younger relatives and unrelated carriers of the same mutation. Hence, the extent of unequal expression of mutated versus wild-type transcript and protein is characteristic for each mutation, implying cis-acting regulatory mechanisms. Bioinformatics suggest mRNA stability or splicing effectors to be affected by certain mutations. Intriguingly, we observed a correlation between disease expression and fraction of mutated mRNA and protein. This strongly suggests that mutation-specific allelic imbalance represents a new pathogenic factor for FHC

Generational Association Studies of Dopaminergic Genes in Reward Deficiency Syndrome (RDS) Subjects: Selecting Appropriate Phenotypes for Reward Dependence Behaviors

Abnormal behaviors involving dopaminergic gene polymorphisms often reflect an insufficiency of usual feelings of satisfaction, or Reward Deficiency Syndrome (RDS). RDS results from a dysfunction in the “brain reward cascade,” a complex interaction among neurotransmitters (primarily dopaminergic and opioidergic). Individuals with a family history of alcoholism or other addictions may be born with a deficiency in the ability to produce or use these neurotransmitters. Exposure to prolonged periods of stress and alcohol or other substances also can lead to a corruption of the brain reward cascade function. We evaluated the potential association of four variants of dopaminergic candidate genes in RDS (dopamine D1 receptor gene [DRD1]; dopamine D2 receptor gene [DRD2]; dopamine transporter gene [DAT1]; dopamine beta-hydroxylase gene [DBH]). Methodology: We genotyped an experimental group of 55 subjects derived from up to five generations of two independent multiple-affected families compared to rigorously screened control subjects (e.g., N = 30 super controls for DRD2 gene polymorphisms). Data related to RDS behaviors were collected on these subjects plus 13 deceased family members. Results: Among the genotyped family members, the DRD2 Taq1 and the DAT1 10/10 alleles were significantly (at least p < 0.015) more often found in the RDS families vs. controls. The TaqA1 allele occurred in 100% of Family A individuals (N = 32) and 47.8% of Family B subjects (11 of 23). No significant differences were found between the experimental and control positive rates for the other variants. Conclusions: Although our sample size was limited, and linkage analysis is necessary, the results support the putative role of dopaminergic polymorphisms in RDS behaviors. This study shows the importance of a nonspecific RDS phenotype and informs an understanding of how evaluating single subset behaviors of RDS may lead to spurious results. Utilization of a nonspecific “reward” phenotype may be a paradigm shift in future association and linkage studies involving dopaminergic polymorphisms and other neurotransmitter gene candidates

10Kin1day: A Bottom-Up Neuroimaging Initiative.

We organized 10Kin1day, a pop-up scientific event with the goal to bring together neuroimaging groups from around the world to jointly analyze 10,000+ existing MRI connectivity datasets during a 3-day workshop. In this report, we describe the motivation and principles of 10Kin1day, together with a public release of 8,000+ MRI connectome maps of the human brain

Transmission of BSE to non human primates as a model for the variant Creutzfeldt-Jakob disease (vCJD) in humans

Die EU-Studie QLK1-CT-2002-01096 soll im Tiermodell das Risiko für den Menschen ermitteln, durch den Konsum von BSE-kontaminierten Nahrungsmitteln, an der varianten Creutzfeldt-Jakob Erkrankung (vCJD) zu erkranken. Als Tiermodell für den Menschen dienen Javaneraffen (Macaca fascicularis). In der Studie wurden Gruppen von je 6 nicht humanen Primaten intracerebral oder oral mit Verdünnungsreihen aus Hirnhomogenat BSE-infizierter Rinder infiziert.In dieser Arbeit konnte nachgewiesen werden, dass 6/6 der mit 50 mg BSE-Hirnhomogenat infizierten Javaneraffen an einer Prionenerkrankung erkrankten. Im Vergleich mit verschiedenen humanen CJD-Subtypen konnte diese als vCJD identifiziert werden. Diese Ergebnisse bestätigen BSE-Material als Auslöser für vCJD. Zusätzlich wurden bei 2/6 Tieren Charakteristika festgestellt, die erstmals auf einen zweiten Prionenstamm bei vCJD hinweisen. Während der Typisierung der Prionenstämme konnten jedoch auch die Grenzen der gängigen Methoden nachgewiesen werden. Über die konformationelle Stabilität des Prion Proteins gegenüber einer Denaturierung mit Guanidinium Hydrochlorid wurden in demselben Tier abhängig von der Hirnregion unterschiedliche Stämme nachgewiesen. In bisher veröffentlichten Studien wurde ein Vergleich der Stabilität als Marker für Prionenstämme zumeist unabhängig von der Hirnregion untersucht. Ein solcher Vergleich ist nach den Ergebnissen dieser Arbeit nicht aussagekräftig.Der zweite Schwerpunkt der Arbeit zielte auf die Suche nach neuen Markern für Prionenerkrankungen. Neben einer Vielzahl von ontogenetischen und zellulären Funktionen wurde in den letzten Jahren das Potential von mikroRNAs (miRNAs) deutlich, als Marker für verschiedene Erkrankungen zu dienen. Tatsächlich konnten in dieser Arbeit drei miRNAs identifiziert werden, die im Zusammenhang mit simianem vCJD hochreguliert waren. Für eine miRNA wurde dies auch für die humane sporadische CJD untersucht und bestätigt. Über eine Analyse der potentiellen Zielgene konnte eine Verbindung der miRNAs zu neurodegenerativen Erkrankungen, wie der Spinozerebralen Ataxie Typ 2 und Chorea Huntington aufgezeigt werden

Asynchronous Onset of Clinical Disease in BSE-Infected Macaques

To estimate the effect of the variability of prion disease onset on primary bovine spongiform encephalopathy transmission to humans, we studied 6 cynomolgus macaques. The preclinical incubation period was significantly prolonged in 2 animals, implying that onset of variant Creutzfeldt-Jacob disease in humans could be more diverse than previously expected