Analysis of repetitive element expression in the blood and skin of patients with Parkinson’s disease identifies differential expression of satellite elements

Repetitive elements (RE) constitute the majority of the human genome and have a range of functions both structural and regulatory on genomic function and gene expression. RE overexpression has been observed in several neurodegenerative diseases, consistent with the observation of aberrant expression of RE posing a mutagenic threat. Despite reports that associate RE expression with PD no study has comprehensively analysed the role of these elements in the disease. This study presents the first genome-wide analysis of RE expression in PD to date. Analysis of RNA-sequencing data of 12 PD patients and 12 healthy controls identified tissue-specific expression differences and more significantly, differential expression of four satellite elements; two simple satellite III (repName = CATTC_n and _GAATG_n) a high-copy satellite II (HSATII) and a centromeric satellite (ALR_Alpha) in the blood of PD patients. In support of the growing body of recent evidence associating REs with neurodegenerative disease, this study highlights the potential importance of characterization of RE expression in such diseases

Serum Amyloid Alpha Is Downregulated in Peripheral Tissues of Parkinson’s Disease Patients

We report the changed levels of serum amyloid alpha, an immunologically active protein, in Parkinson’s disease (PD) patients’ peripheral tissues. We have previously shown that Saa-1 and -2 (serum amyloid alpha-1,-2, genes) were among the top downregulated genes in PD patients’ skin, using whole-genome RNA sequencing. In the current study, we characterized the gene and protein expression profiles of skin and blood samples from patients with confirmed PD diagnosis and age/sex matched controls. qRT-PCR analysis of PD skin demonstrated downregulation of Saa-1 and -2 genes in PD patients. However, the lowered amount of protein could not be visualized using immunohistochemistry, due to low quantity of SAA (Serum Amyloid Alpha, protein) in skin. Saa-1 and -2 expression levels in whole blood were below detection threshold based on RNA sequencing, however significantly lowered protein levels of SAA1/2 in PD patients’ serum were shown with ELISA, implying that SAA is secreted into the blood. These results show that SAA is differentially expressed in the peripheral tissues of PD patients

Bi-allelic variants in TSPOAP1, encoding the active zone protein RIMBP1, cause autosomal recessive dystonia

Dystonia is a debilitating hyperkinetic movement disorder, which can be transmitted as a monogenic trait. Here, we describe homozygous frameshift, nonsense and missense variants in TSPOAP1, encoding the active zone RIM-binding protein 1 (RIMBP1), as a novel genetic cause of autosomal recessive dystonia in seven subjects from three unrelated families. Subjects carrying loss-of-function variants presented with juvenile-onset progressive generalized dystonia, associated with intellectual disability and cerebellar atrophy. Conversely, subjects carrying a pathogenic missense variant (p.Gly1808Ser) presented with isolated adult-onset focal dystonia. In mice, complete loss of RIMBP1, known to reduce neurotransmission, led to motor abnormalities reminiscent of dystonia, decreased Purkinje cell dendritic arborization, and reduced numbers of cerebellar synapses. In vitro analysis of the p.Gly1808Ser variant showed larger spike-evoked calcium transients and enhanced neurotransmission, suggesting that RIMBP1-linked dystonia can be caused by either reduced or enhanced rates of spike-evoked release in relevant neural networks. Our findings establish a direct link between dysfunction of the presynaptic active zone and dystonia and highlight the critical role played by well-balanced neurotransmission in motor control and disease pathogenesis

Biallelic variants in TSPOAP1, encoding the active-zone protein RIMBP1, cause autosomal recessive dystonia.

Dystonia is a debilitating hyperkinetic movement disorder, which can be transmitted as a monogenic trait. Here, we describe homozygous frameshift, nonsense, and missense variants in TSPOAP1, which encodes the active-zone RIM-binding protein 1 (RIMBP1), as a genetic cause of autosomal recessive dystonia in 7 subjects from 3 unrelated families. Subjects carrying loss-of-function variants presented with juvenile-onset progressive generalized dystonia, associated with intellectual disability and cerebellar atrophy. Conversely, subjects carrying a pathogenic missense variant (p.Gly1808Ser) presented with isolated adult-onset focal dystonia. In mice, complete loss of RIMBP1, known to reduce neurotransmission, led to motor abnormalities reminiscent of dystonia, decreased Purkinje cell dendritic arborization, and reduced numbers of cerebellar synapses. In vitro analysis of the p.Gly1808Ser variant showed larger spike-evoked calcium transients and enhanced neurotransmission, suggesting that RIMBP1-linked dystonia can be caused by either reduced or enhanced rates of spike-evoked release in relevant neural networks. Our findings establish a direct link between dysfunction of the presynaptic active zone and dystonia and highlight the critical role played by well-balanced neurotransmission in motor control and disease pathogenesis

Parkinson’s disease as a multisystem disorder: Whole transcriptome study in Parkinson’s disease patients’ skin and blood–finding the pathomechanistic link

Background and aims: Next to characteristic motor triad of Parkinson’s Disease (PD) due to loss of nigrostriatal neurons, more symptoms associated with non-neuronal tissues are emerging. Little is known about the molecular alterations underlying dermatologic issues or epidemiologic associations like increased incidence of melanoma in PD. The aim is to give an overview of the altered gene expression profiles of PD skin and blood using the novel method of RNA Sequencing. Networks of different genes are analyzed to map affected pathways that contribute to pathomolecular mechanism of PD in the periphery. Methods: Whole transcriptomic profiling of 12+12 idiopathic PD patients’ and matched controls’ skin biopsies and venous whole blood was performed with highthroughput RNA-sequencing analysis. Followingly, pathway analysis of differentially changed gene expressions was performed. The results were validated using RT-qPCR. Results: PD skin RNA-Seq resulted in a large collection of over 1000 differentially expressed genes, among which a clear pattern of global downregulation appeared. In blood, the differential changes were more subtle, blood being a heterogenous tissue. Pathways associated with mitochondrial metabolism and protein degradation by the ubiquitin-proteasome system were dysregulated in both. Conclusion: The concordance of these results with previous gene expression profiling studies demonstrate that the molecular alterations in PD leading to neurodegeneration in the CNS are systemic and manifest also in peripheral tissues. Major affected pathways include dysfunction in protein metabolism, mitochondrial dysfunction and impaired immune system. Homeostatic imbalance in the skin can lead to increased susceptibility to mutagenic hazards and provide a possible molecular link between melanoma and PD

Transcriptomic profiles in Parkinson’s disease

Transcriptomics in Parkinson’s disease offers insights into the pathogenesis of Parkinson’s disease but obtaining brain tissue has limitations. In order to bypass this issue, we profile and compare differentially expressed genes and enriched pathways (KEGG) in two peripheral tissues (blood and skin) of 12 Parkinson’s disease patients and 12 healthy controls using RNA-sequencing technique and validation with RT-qPCR. Furthermore, we compare our results to previous Parkinson’s disease post mortem brain tissue and blood results using the robust rank aggregation method. The results show no overlapping differentially expressed genes or enriched pathways in blood vs. skin in our sample sets (25 vs. 1068 differentially expressed genes with an FDR ≤ 0.05; 1 vs. 9 pathways in blood and skin, respectively). A meta-analysis from previous transcriptomic sample sets using either microarrays or RNA-Seq yields a robust rank aggregation list of cortical gene expression changes with 43 differentially expressed genes; a list of substantia nigra changes with 2 differentially expressed genes and a list of blood changes with 1 differentially expressed gene being statistically significant at FDR ≤ 0.05. In cortex 1, KEGG pathway was enriched, four in substantia nigra and two in blood. None of the differentially expressed genes or pathways overlap between these tissues. When comparing our previously published skin transcription analysis, two differentially expressed genes between the cortex robust rank aggregation and skin overlap. In this study, for the first time a meta-analysis is applied on transcriptomic sample sets in Parkinson’s disease. Simultaneously, it explores the notion that Parkinson’s disease is not just a neuronal tissue disease by exploring peripheral tissues. The comparison of different Parkinson’s disease tissues yields surprisingly few significant differentially expressed genes and pathways, suggesting that divergent gene expression profiles in distinct cell lineages, metabolic and possibly iatrogenic effects create too much transcriptomic noise for detecting significant signal. On the other hand, there are signs that point towards Parkinson’s disease-specific changes in non-neuronal peripheral tissues in Parkinson’s disease, indicating that Parkinson’s disease might be a multisystem disorder

Adapting The Sniffin' Sticks Olfactory Test To Diagnose Parkinson's Disease In Estonia

The aim of the study was to develop a culturally adapted translation of the 12-item smell identification test from Sniffin' Sticks (SS-12) for the Estonian population in order to help diagnose Parkinson's disease (PD). Methods: A standard translation of the SS-12 was created and 150 healthy Estonians were questioned about the smells used as response options in the test. Unfamiliar smells were replaced by culturally familiar options. The adapted SS-12 was applied to 70 controls in all age groups, and thereafter to 50 PD patients and 50 age- and sex-matched controls. Results: 14 response options from 48 used in the SS-12 were replaced with familiar smells in an adapted version, in which the mean rate of correct response was 87% (range 73-99) compared to 83% with the literal translation (range 50-98). In PD patients, the average adapted SS-12 score (5.4/12) was significantly lower than in controls (average score 8.9/12), p<0.0001. A multiple linear regression using the score in the SS-12 as the outcome measure showed that diagnosis and age independently influenced the result of the SS-12. A logistic regression using the SS-12 and age as covariates showed that the SS-12 (but not age) correctly classified 79.0% of subjects into the PD and control category, using a cut-off of <7 gave a sensitivity of 76% and specificity of 86% for the diagnosis of PD. Conclusions: The developed SS-12 cultural adaption is appropriate for testing olfaction in Estonia for the purpose of PD diagnosis. © 2014 Elsevier Ltd.208830833Soudrya, Y., Lemogne, C., Malinvaud, D., Consoli, S.-M., Bonfils, P., Olfactory system and emotion: common substrates (2011) Eur Ann Otorhinolaryngol Head Neck Dis, 128, pp. 18-23Kovács, T., Mechanisms of olfactory dysfunction in aging and neurodegenerative disorders (2004) Ageing Res Rev, 3, pp. 215-232Chaudhuri, K.R., Odin, P., The challenge of non-motor symptoms in Parkinson's disease (2010) Prog Brain Res, 184, pp. 325-341Bohnen, N.I., Studenski, S.A., Constantine, G.M., Moore, R.Y., Diagnostic performance of clinical motor and non-motor tests of Parkinson disease: a matched case-control study (2008) Eur J Neurol, 15 (7), pp. 685-691Silveira-Moriyama, L., de Jesus Carvalho, M., Katzenschlager, R., Petrie, A., Ranvaud, R., Barbosa, E.R., The use of smell identification tests in the diagnosis of Parkinson's disease in Brazil (2008) Mov Disord, 23, pp. 2328-2334Silveira-Moriyama, L., Schwingenschuh, P., O'Donnell, A., Schneider, S.A., Mir, P., Carrillo, F., Olfaction in patients with suspected Parkinsonism and scans without evidence of dopaminergic deficit (SWEDDs) (2009) JNeurol Neurosurg Psychiatry, 80 (7), pp. 744-748Doty, R.L., Shaman, P., Kimmelman, C., Dann, M.S., University of Pennsylvania Smell Identification Test: a rapid quantitative olfactory function test for the clinic (1984) Laryngoscope, 94, pp. 176-178Doty, R.L., Marcus, A., Lee, W.W., Development of the 12-Item cross-cultural smell identification test (CC-SIT) (1996) Laryngoscope, 106, pp. 353-356Hummel, T., Konnerth, C.G., Rosenheim, K., Kobal, G., Screening of olfactory function using a 4 minute odor identification test: reliability, normative data, and investigations in patients with olfactory loss (2001) Ann Otol Rhinol Laryngol, 110, pp. 976-981Nordin, S., Bramerson, A., Liden, E., Bende, M., The Scandinavian odor-identification test: development, reliability, validity and normative data (1998) Acta Otolaryngol, 118, pp. 226-234Simmen, D., Briner, H.R., Hess, K., Screeningtest des Geruchssinnes mit Riechdiskette (1999) Laryngorhinootologie, 78, pp. 125-130Hummel, C., Zucco, G.M., Iannilli, E., Maboshe, W., Landis, B.N., Hummel, T., OLAF: standardization of international olfactory tests (2012) Eur Arch Otorhinolaryngol, 269 (3), pp. 871-880Cho, J.H., Jeong, Y.S., Lee, Y.J., Hong, S.C., Yoon, J.H., Kim, J.K., The Korean version of the Sniffin' stick (KVSS) test and its validity in comparison with the cross-cultural smell identification test (CC-SIT) (2009) Auris Nasus Larynx, 36, pp. 280-286Shu, C.H., Yuan, B.C., Lin, S.H., Lin, C.Z., Cross-cultural application of the "Sniffin' Sticks" odor identification test (2007) Am J Rhinol, 21 (5), pp. 570-573Silveira-Moriyama, L., Sirisena, D., Gamage, P., Ranjanie Gamage, R., Silva, R., Lees, A.J., Adapting the sniffin' sticks to diagnose PD in Sri Lanka (2009) Mov Disord, 24 (8), pp. 1229-1233Mackay-Sim, A., Grant, L., Owen, C., Chant, D., Silburn, P., Australian norms for a quantitative olfactory function test (2004) JClin Neurosci, 11, pp. 874-879Neumann, C., Tsioulos, K., Merkonidis, C., Salam, M., Clark, A., Philpott, C., Validation study of the "Sniffin' Sticks" olfactory test in a British population: a preliminary communication (2012) Clin Otolaryngol, 37 (1), pp. 23-27Eibenstein, A., Fioretti, A.B., Lena, C., Rosati, N., Ottaviano, I., Fusetti, M., Olfactory screening test: experience in 102 Italian subjects (2005) Acta Otorhinolaryngol Ital, 25 (1), pp. 18-22Boesveldt, S., Verbaan, D., Knol, D.L., Visser, M., van Rooden, S.M., van Hilten, J.J., Acomparative study of odor identification and odor discrimination deficits in Parkinson's disease (2008) Mov Disord, 23 (14), pp. 1984-1990Boesveldt, S., de Muinck Keizer, R.J., Knol, D.L., Wolters, E., Berendse, H.W., Extended testing across, not within, tasks raises diagnostic accuracy of smell testing in Parkinson's disease (2009) Mov Disord, 24 (1), pp. 85-90Hummel, T., Kobal, G., Gudziol, H., Mackay-Sim, A., Normative data for the "Sniffin' Sticks" including tests of odor identification, odor discrimination, and olfactory thresholds: an upgrade based on a group of more than 3,000 subjects (2007) Eur Arch Otorhinolaryngol, 264, pp. 237-243Doty, R.L., Olfactory dysfunction in Parkinson disease (2012) Nat Rev Neurol, 8 (6), pp. 329-339Brand, G., Millot, J.L., Sex differences in human olfaction: between evidence and enigma (2001) QJ Exp Psychol B, 54 (3), pp. 259-270Larsson, M., Nilsson, L.G., Olofsson, J.K., Nordin, S., Demographic and cognitive predictors of cued odor identification: evidence from a population-based study (2004) Chem Senses, 29 (6), pp. 547-554Doty, R.L., Cameron, E.L., Sex differences and reproductive hormone influences on human odor perception (2009) Physiol Behav, 97 (2), pp. 213-228Orhan, K.S., Karabulut, B., Keles, N., Deger, K., Evaluation of factors concerning the olfaction using the sniffin' sticks test (2012) Otolaryngol Head Neck Surg, 146 (2), pp. 240-246Frye, R.E., Schwartz, B.S., Doty, R.L., Dose-related effects of cigarette smoking on olfactory function (1990) JAMA, 263, pp. 1233-1236Katotomichelakis, M., Balatsouras, D., Tripsianis, G., Davris, S., Maroudias, N., Danielides, V., The effect of smoking on the olfactory function (2007) Rhinology, 45, pp. 273-280Bower, J.H., Maraganore, D.M., Peterson, B.J., Ahlskog, J.E., Rocca, W.A., Immunologic diseases, anti-inflammatory drugs, & Parkinson disease: a case-control study (2006) Neurology, 67 (3), pp. 494-496Sedig, L., Leibner, J., Ramjit, A.L., Wu, S.S., Dai, Y., Okun, M.S., Is rhinorrhea an under-recognized intrinsic symptom of Parkinson disease? A prospective pilot study (2010) Int J Neurosci, 120 (4), pp. 258-26

Gene expression in skin of Parkinson’s disease patients

Poster Session

Differences in MDS-UPDRS Scores Based on Hoehn and Yahr Stage and Disease Duration

BACKGROUND: The Movement Disorder Society Unified Parkinson's Disease Rating Scale (MDS‐UPDRS) is a newly developed tool to assess Parkinson's disease (PD). Changes in scores on the scale over the course of PD, including increasing disease duration and Hoehn and Yahr (HY) stages, have not been described. The objectives of this study were to analyze MDS‐UPDRS scores on Parts I through IV and their differences based on HY stage and disease duration in a large cohort of patients with PD. METHODS: For this cross‐sectional study, demographic data and MDS‐UPDRS scores were collected, including HY stage. Subscores on MDS‐UPDRS Parts I through IV were analyzed using 1‐way analyses of variance for each HY stage and in 5‐year increments of disease duration. Part III (motor assessment) scores were analyzed separately for on and off states. RESULTS: The mean age of the 3206 patients was 65.8 ± 10.6 years, 53.3% were men, the mean disease duration was 11.5 ± 4.6 years, and the median HY stage was 2 (range, 0–5); 2156 patients were examined in an on state and 987 were examined in an off state. Scores for all MDS‐UPDRS parts increased significantly through HY stages 1 through 5, with an average increase of 3.8, 7.7, 14.6, and 2.0 points consecutively for parts I through IV, respectively. For the 5‐year increments of disease duration, MDS‐UPDRS subscores increased by an average of 1.6, 3.3, 4.2, and 1.4 points consecutively for parts I through IV, respectively. This increase was significant only during the first 15 years of disease for all 4 parts, including part III scores evaluated in both on and off states. CONCLUSIONS: MDS‐UPDRS scores for all 4 parts increase significantly with every HY stage and also with 5‐year increments of disease duration in the first 15 years of the disease