Astrocytes Differentiated from LRRK2-I1371V Parkinson's-Disease-Induced Pluripotent Stem Cells Exhibit Similar Yield but Cell-Intrinsic Dysfunction in Glutamate Uptake and Metabolism, ATP Generation, and Nrf2-Mediated Glutathione Machinery.

Owing to the presence of multiple enzymatic domains, LRRK2 has been associated with a diverse set of cellular functions and signaling pathways. It also has several pathological mutant-variants, and their incidences show ethnicity biases and drug-response differences with expression in dopaminergic-neurons and astrocytes. Here, we aimed to assess the cell-intrinsic effect of the LRRK2-I1371V mutant variant, prevalent in East Asian populations, on astrocyte yield and biology, involving Nrf2-mediated glutathione machinery, glutamate uptake and metabolism, and ATP generation in astrocytes derived from LRRK2-I1371V PD patient iPSCs and independently confirmed in LRRK2-I1371V-overexpressed U87 cells. Astrocyte yield (GFAP-immunopositive) was comparable between LRRK2-I1371V and healthy control (HC) populations; however, the astrocytic capability to mitigate oxidative stress in terms of glutathione content was significantly reduced in the mutant astrocytes, along with a reduction in the gene expression of the enzymes involved in glutathione machinery and nuclear factor erythroid 2-related factor 2 (Nrf2) expression. Simultaneously, a significant decrease in glutamate uptake was observed in LRRK2-I1371V astrocytes, with lower gene expression of glutamate transporters SLC1A2 and SLC1A3. The reduction in the protein expression of SLC1A2 was also directly confirmed. Enzymes catalyzing the generation of γ glutamyl cysteine (precursor of glutathione) from glutamate and the metabolism of glutamate to enter the Krebs cycle (α-ketoglutaric acid) were impaired, with significantly lower ATP generation in LRRK2-I1371V astrocytes. De novo glutamine synthesis via the conversion of glutamate to glutamine was also affected, indicating glutamate metabolism disorder. Our data demonstrate for the first time that the mutation in the LRRK2-I1371V allele causes significant astrocytic dysfunction with respect to Nrf2-mediated antioxidant machinery, AT -generation, and glutamate metabolism, even with comparable astrocyte yields.

Plasma microRNAs as a Potential Biomarker for Identification of Progressive Supranuclear Palsy.

Progressive supranuclear palsy (PSP) is the second most common Parkinsonian disorder with complex etiology. The underlying molecular mechanism of PSP pathogenesis remains unclear. The present study aims to find the feasibility of using plasma miRNAs as novel biomarkers. Plasma-focused qPCR panels were used for microRNA profiling and identified differentially expressed microRNAs in PSP compared to controls. The DIANA-miRPath v3.0 was used to perform KEGG pathway analysis. We then confirmed the expression of selected candidates by RT-qPCR and their clinical utility was assessed by ROC analysis. Profiling data revealed 28 differentially expressed microRNAs in PSP. Five overexpressed miRNAs were selected for further analysis. The KEGG pathway analysis revealed 48 high-risk pathways. The study revealed that as a single marker-miR-19b-3p, miR-33a-5p, miR-130b-3p, miR-136-3p, and miR-210-3p had a specificity of 64.71%, 82.35%, 68.75%, 82.35%, and 70.59% at sensitivity 77.78%, 77.78%, 66.67%, 73.33%, and 66.67%, respectively. The result suggests that circulating plasma miRNAs were altered in PSP compared to control. The findings of this study may provide potential biomarkers and pathways associated with PSP. Further large-scale validation studies are required to confirm the same.

Speech Characteristics of Patients with Parkinson's Disease-Does Dopaminergic Medications Have a Role?

Objective The present study aimed to investigate the effects of dopaminergic medication on voice, speech motor functions, and motor impairment in patients with Parkinson's disease (PD). Materials and Methods Twenty-five individuals (16 males and 9 females) with PD underwent comprehensive assessment of voice, speech, and motor functions in levodopa medication ON and medication OFF conditions. Age- and gender-matched healthy controls were recruited to compare speech and acoustic parameters. Multi-Dimensional Voice Program (MDVP) from Computerized Speech Laboratory (Model: 4500) was utilized for acoustic analysis of voice and the Voice Handicap Index (VHI) for the self-assessment of vocal function. Frenchay Dysarthria Assessment (FDA-2) and Unified Parkinson's Disease Rating Scale-III (UPDRS III) were used to evaluate speech motor and motor functions, respectively. Statistical Analysis The mean and standard deviation were used as descriptive statistics measures. Raw scores were obtained for FDA-2, DRS, VHI, MDVP parameters, and UPDRS-III in either medication condition. The Wilcoxon signed-rank test was performed to determine the statistical significance of the above measures in both genders across the medication conditions. Spearman's rank correlation coefficient was used to determine the relationship between motor speech function and motor impairment and between VHI and MDVP parameters across both medication conditions. The interrater reliability rating was established using Cohen's kappa. Results An improvement in lip and laryngeal functioning was found in the medication ON over medication OFF state in both males and females with PD. A few frequency and amplitude-related measures improved in the medication-ON state over the medication-OFF state. UPDRS-III scores reduced from the OFF state to the ON state, and no change in dysarthria severity or VHI was found in either gender or medication condition. No correlation was found between speech motor function and motor function or between VHI and acoustic parameters of voice in either medication condition. Conclusions Improvement in motor symptoms with levodopa was predominantly observed when compared with the minor improvements in a few aspects of speech motor function and vocal parameters. The results of this study suggest the need for speech therapy as a nonpharmacological treatment method for speech impairments in PD.

Assessment of Sleep Spindle Density among Genetically Positive Spinocerebellar Ataxias Types 1, 2, and 3 Patients.

OBJECTIVE: The effect of thalamic degeneration in patients with spinocerebellar ataxias (SCA) and sleep spindle (SS) abnormalities has not been studied so far, although there is a strong association between these disorders. This study was done to evaluate and compare the SS densities (SSDs) of genetically proven autosomal dominant SCA1, SCA2 and SCA3 patients with controls. METHODS: Prospectively and genetically confirmed cases of SCA and controls were recruited. Patients were assessed clinically, were evaluated with sleep questionnaires and an overnight polysomnography was performed. SSDs were analyzed using neuroloop gain plugin of Polyman version 1.15 software. RESULTS: Eighteen patients of SCA1 (n = 6), SCA2 (n = 5), SCA3 (n = 7) and 6 controls were recruited in our study. The mean age of SCA1 patients was 39.2 ± 5.4, of SCA2 patients was 30.8 ± 9.5 and of SCA3 patients was 35.4 ± 6.4 years. The mean duration of illness in SCA1 was 4.7 ± 1.7 years, in SCA2 it was 4.3 ± 4.4 years and in SCA3 it was 5 ± 2.3 years. The median SSD values (percentage loop gain) during stage 2 of non-rapid eye movement sleep were 16.9% in SCA1, 0% in SCA2, 1.2% in SCA3 and 59.5% in controls. There was a significant difference in SSD values in SCA2 (p = 0.04), SCA3 (p = 0.02) patients and controls. CONCLUSION: SSDs were significantly decreased in patients with SCA, which is a novel finding. This is likely due to the "thalamic switch" disruption, observed as reduced SSDs in SCA2 and SCA3. Sleep spindle deficits could act as one of the biomarkers of ongoing neurodegeneration in the thalamic circuitry of SCA patients.

Handwriting Analysis in Parkinson's Disease: Current Status and Future Directions.

BACKGROUND: The majority of patients with Parkinson's disease (PD) have handwriting abnormalities. Micrographia (abnormally small letter size) is the most commonly reported and easily detectable handwriting abnormality in patients with PD. However, micrographia is perhaps the tip of the iceberg representing the handwriting abnormalities in PD. Digitizing tablet technology, which has evolved over the last 2 decades, has made it possible to study the pressure and kinematic features of handwriting. This has resulted in a surge of studies investigating graphomotor impairment in patients with PD. METHODS: The objectives of this study were to review the evolution of the kinematic analysis of handwriting in PD and to provide an overview of handwriting abnormalities observed in PD along with future directions for research in this field. Articles for review were searched from the PubMed and SCOPUS databases. RESULTS: Digitizing tablet technologies have resulted in a shift of focus from the analysis of only letter size to the analysis of several kinematic features of handwriting. Studies based on the kinematic analysis of handwriting have revealed that patients with PD may have abnormalities in velocity, fluency, and acceleration in addition to micrographia. The recognition of abnormalities in several kinematic parameters of handwriting has given rise to the term PD dysgraphia. In addition, certain kinematic properties potentially may be helpful in distinguishing PD from other parkinsonian disorders. CONCLUSION: The journey from micrographia to PD dysgraphia is indeed a paradigm shift. Further research is warranted to gain better insight into the graphomotor impairments in PD and their clinical implications.