Biology subjects

Ferreira, A. F.

Publications and source records attributed to Ferreira, A. F..

4 recordsLinked to original sources

bioRxiv2023

Blood DDIT4 and TRIM13 transcript levels mark the early stages of Machado-Joseph disease

Machado-Joseph disease (MJD) is a rare late-onset polyglutamine neurodegenerative disease caused by the expansion of a CAG repeat in the ATXN3 gene encoding the ataxin- 3 (ATXN3) protein. Several studies have identified changes in the abundance of select transcripts and proteins in blood samples of MJD mutation carriers. Here, we aimed to: 1) identify blood transcriptional changes that could be potential biomarkers of MJD from preclinical to symptomatic disease stages; 2) correlate levels of differentially expressed transcripts in blood of MJD carriers with demographic, genetics, and clinical features; and 3) evaluate whether the identified differential abundance of selected transcripts in blood of MJD subjects is preserved in post-mortem brains of MJD patients. Using real- time quantitative PCR, we observed consistent dysregulation of DDIT4, TRIM13 and P2RY13 transcript levels in blood samples of MJD subjects from different disease stages (from preclinical to symptomatic), and of patients from two cohorts with different backgrounds (Azores and Brazil). Importantly, combined blood DDIT4 and TRIM13 transcript levels display a very high accuracy to discriminate MJD carriers in preclinical stage, early-stage patients, and patients with more than five years of disease duration from respective controls (AUC=1.00, AUC=0.96, and AUC=0.90, respectively). Moreover, in the combined group of Azorean and Brazilian patients, blood levels of P2RY13 transcript correlate with age at onset, and abundance of DDIT4 and TRIM13 transcripts correlate with the CAG repeat size of the expanded ATXN3 allele. In the subgroup of early-stage Azorean patients, blood levels of TRIM13 transcripts correlate with age at disease onset. Interestingly, abundance of DDIT4, TRIM13 and P2RY13 proteins is also altered in brains of MJD patients. In summary, this work shows that blood DDIT4 and TRIM13 transcript levels are potentially blood-based biomarkers of MJD of special usefulness in marking preclinical and early stages of the disease, and points for common dysregulated processes involving DDIT4, TRIM13 and P2RY13 in the blood and the brain of MJD subjects.

neuroscience↗

bioRxiv2023

Blood and cerebellar abundance of ATXN3 splice variants in spinocerebellar ataxia type 3/Machado-Joseph disease

BackgroundSpinocerebellar ataxia type 3 (SCA3)/Machado-Joseph disease (MJD) is an autosomal dominant polyglutamine disease. SCA3/MJD causative gene, ATXN3, is known to undergo alternative splicing (AS) and 54 transcripts are currently annotated. Differences in the toxicity of ataxin-3 protein isoforms, harbouring on its C-terminus two or three ubiquitin interacting motifs (UIMs), were previously uncovered, raising the hypothesis that specific ATXN3 splice variants play key roles in promoting the selective toxicity displayed in SCA3/MJD. MethodsUsing RNA-seq datasets we identified and determined the abundance of annotated ATXN3 transcripts in blood (n=60) and cerebellum (n=12) of SCA3/MJD subjects and controls. ResultsGlobally, the number and the abundance of individual ATXN3 transcripts were higher in the cerebellum than in the blood. While the most abundant transcript in the cerebellum was a protein with a coding sequence not defined of unknown function (ATXN3-208), the transcript with the highest abundance in blood was the reference transcript (ATXN3-251) which translates into an ataxin-3 isoform harboring three UIMs. Noteworthy, the abundance of ATXN3-251 and ATXN3-214, two out of the four transcripts that encode full-length ataxin-3 protein isoforms but differ in the C-terminus were strongly related with tissue expression specificity: ATXN3-251 (3UIM) was expressed in blood 50-fold more than in cerebellum, whereas ATXN3-214 (2UIM) was expressed in the cerebellum 20-fold more than in blood. ConclusionsThese findings provide new insights into the elucidation of ATXN3 AS in different tissues, contributing for a better understanding of SCA3/MJD pathogenesis and providing information for the development of future effective ATXN3 mRNA-lowering therapies.

genetics↗

bioRxiv2023

Tissue-specific vulnerability to apoptosis in Machado-Joseph disease

Machado-Joseph disease (MJD) is a dominant neurodegenerative disease caused by an expanded CAG repeat in the ATXN3 gene encoding the ataxin-3 protein. Several cellular processes, including transcription and apoptosis, are disrupted in MJD. To gain further insights into the extent of dysregulation of mitochondrial apoptosis in MJD, and to evaluate if expression alterations of specific apoptosis genes/proteins could be used as transcriptional biomarkers of disease, the levels of BCL2, BAX and TP53 and the BCL2/BAX ratio, an indicator of susceptibility to apoptosis, were assessed in blood and post-mortem brain samples from MJD subjects and MJD transgenic mice and controls. While patients show reduced levels of blood BCL2 transcripts, this measurement displays low accuracy to discriminate patients from matched controls. However, increased levels of blood BAX transcripts and decreased BCL2/BAX ratio are associated with earlier onset, indicating a possible association with MJD pathogenesis. Post-mortem MJD brains show increased BCL2/BAX transcript ratio in the dentate cerebellar nucleus (DCN) and increased BCL2/BAX insoluble protein ratio in the DCN and pons, suggesting that in these regions, severely affected by degeneration in MJD, cells show signs of apoptosis resistance. Interestingly, a follow-up study of 18 patients further shows that blood BCL2 and TP53 transcript levels increase over time in MJD patients. Furthermore, while the similar levels of blood BCL2, BAX, and TP53 transcripts observed in preclinical subjects and controls is mimicked by pre-symptomatic MJD mice, the expression profile of these genes in patient brains is partially replicated by symptomatic MJD mice. Globally, our findings indicate that there is tissue-specific vulnerability to apoptosis in MJD subjects and that this tissue dependent behavior is partially replicated in a MJD mouse model.

neuroscience↗

bioRxiv2022

Altered retinal structure and function in Spinocerebellar ataxia type 3

Spinocerebellar ataxia type 3 is an autosomal dominant neurodegenerative disorder caused by expansion of a polyglutamine (polyQ)-encoding CAG repeat in the ATXN3 gene. Because the ATXN3 protein regulates photoreceptor ciliogenesis and phagocytosis, we aimed to explore whether expanded polyQ ATXN3 impacts retinal function and integrity in SCA3 patients and transgenic mice. We evaluated the retinal structure and function in five patients with Spinocerebellar ataxia type 3 and in a transgenic mouse model of this disease (YACMJD84.2, Q84) using, respectively, optical coherence tomography (OCT) and electroretinogram (ERG). We further determined in the transgenic mice: a) the retinal expression pattern of ATXN3 and assessed the distribution of cones and rods by immunofluorescence (IF); and b) the retinal ultrastructure by transmission electron microscopy (TEM). Some patients with Spinocerebellar ataxia type 3 in our cohort revealed: i) reduced central macular thickness indirectly correlated with disease duration; ii) decreased thickness of the macula and the ganglion cell layer, and reduced macula volume inversely correlated with disease severity (SARA score); and iii) electrophysiological dysfunction of cones, rods, and inner retinal cells. Transgenic mice replicated the human OCT and ERG findings with aged homozygous Q84/Q84 mice showing a stronger phenotype accompanied by further thinning of the outer nuclear layer and photoreceptor layer and highly reduced cone and rod activities, thus supporting severe retinal dysfunction in these mice. In addition, Q84 mice showed progressive accumulation of ATXN3-positive aggregates throughout several retinal layers and depletion of cones alongside the disease course. TEM analysis of aged Q84/Q84 mouse retinas supported the IF ATXN3 aggregation findings by revealing the presence of high number of negative electron dense puncta in ganglion cells, inner plexiform and inner nuclear layers, and further showed thinning of the outer plexiform layer, thickening of the retinal pigment epithelium and elongation of apical microvilli. Our results indicate that retinal alterations detected by non-invasive eye examination using OCT and ERG could represent a biological marker of disease progression and severity in patients with Spinocerebellar ataxia type 3.

neuroscience↗