Rare and Disabling Movement Disorders: An Indian Experience

Recent decades have seen exciting developments in the field of movement disorders. These include identification of rare clinical syndromes and use of technological advances to understand their pathogenesis. Three such disorders are discussed here. The description of the Uner Tan syndrome from Turkey and surrounding regions provoked research into the controversial field of genetically induced devolution. Such cases with few additional findings have now been described from India. Sepiapterin reductase deficiency is a rare treatable autosomal recessive form of dopa responsive dystonia. Indian literature has recently added five confirmed cases to the international database. Such cases are eminently treatable. Successful application of modern technology in understanding the pathogenesis of progressive neurodegenerative disorder has been highlighted in the section on hereditary spastic paraplegias. Hitherto undescribed subcellular organelle transport defects and their potential rectification with known drugs have been demonstrated raising hopes for their cure

Reduced acetylated α-tubulin in SPAST hereditary spastic paraplegia patient PBMCs

HSP-SPAST is the most common form of hereditary spastic paraplegia (HSP), a neurodegenerative disease causing lower limb spasticity. Previous studies using HSP-SPAST patient-derived induced pluripotent stem cell cortical neurons have shown that patient neurons have reduced levels of acetylated α-tubulin, a form of stabilized microtubules, leading to a chain of downstream effects eventuating in increased vulnerability to axonal degeneration. Noscapine treatment rescued these downstream effects by restoring the levels of acetylated α-tubulin in patient neurons. Here we show that HSP-SPAST patient non-neuronal cells, peripheral blood mononuclear cells (PBMCs), also have the disease-associated effect of reduced levels of acetylated α-tubulin. Evaluation of multiple PBMC subtypes showed that patient T cell lymphocytes had reduced levels of acetylated α-tubulin. T cells make up to 80% of all PBMCs and likely contributed to the effect of reduced acetylated α-tubulin levels seen in overall PBMCs. We further showed that mouse administered orally with increasing concentrations of noscapine exhibited a dose-dependent increase of noscapine levels and acetylated α-tubulin in the brain. A similar effect of noscapine treatment is anticipated in HSP-SPAST patients. To measure acetylated α-tubulin levels, we used a homogeneous time resolved fluorescence technology-based assay. This assay was sensitive to noscapine-induced changes in acetylated α-tubulin levels in multiple sample types. The assay is high throughput and uses nano-molar protein concentrations, making it an ideal assay for evaluation of noscapine-induced changes in acetylated α-tubulin levels. This study shows that HSP-SPAST patient PBMCs exhibit disease-associated effects. This finding can help expedite the drug discovery and testing process

Whole genome sequencing for the genetic diagnosis of heterogenous dystonia phenotypes

Introduction: Dystonia is a clinically and genetically heterogeneous disorder and a genetic cause is often difficult to elucidate. This is the first study to use whole genome sequencing (WGS) to investigate dystonia in a large sample of affected individuals. Methods: WGS was performed on 111 probands with heterogenous dystonia phenotypes. We performed analysis for coding and non-coding variants, copy number variants (CNVs), and structural variants (SVs). We assessed for an association between dystonia and 10 known dystonia risk variants. Results: A genetic diagnosis was obtained for 11.7% (13/111) of individuals. We found that a genetic diagnosis was more likely in those with an earlier age at onset, younger age at testing, and a combined dystonia phenotype. We identified pathogenic/likely-pathogenic variants in ADCY5 (n = 1), ATM (n = 1), GNAL (n = 2), GLB1 (n = 1), KMT2B (n = 2), PRKN (n = 2), PRRT2 (n = 1), SGCE (n = 2), and THAP1 (n = 1). CNVs were detected in 3 individuals. We found an association between the known risk variant ARSG rs11655081 and dystonia (p = 0.003). Conclusion: A genetic diagnosis was found in 11.7% of individuals with dystonia. The diagnostic yield was higher in those with an earlier age of onset, younger age at testing, and a combined dystonia phenotype. WGS may be particularly relevant for dystonia given that it allows for the detection of CNVs, which accounted for 23% of the genetically diagnosed cases. © 2019 The Author

Patient-Derived Stem Cell Models in SPAST HSP: Disease Modelling and Drug Discovery

Hereditary spastic paraplegia is an inherited, progressive paralysis of the lower limbs first described by Adolph Strümpell in 1883 with a further detailed description of the disease by Maurice Lorrain in 1888. Today, more than 100 years after the first case of HSP was described, we still do not know how mutations in HSP genes lead to degeneration of the corticospinal motor neurons. This review describes how patient-derived stem cells contribute to understanding the disease mechanism at the cellular level and use this for discovery of potential new therapeutics, focusing on SPAST mutations, the most common cause of HSP

Antibody-free targeted proteomics assay for absolute measurement of α-tubulin acetylation

Acetylation of α-tubulin at conserved lysine 40 (K40) amino acid residue regulates microtubule dynamics and controls a wide range of cellular activities. Dysregulated microtubule dynamics characterized by differential α-tubulin acetylation is a hallmark of cancer, neurodegeneration, and other complex disorders. Hence, accurate quantitation of α-tubulin acetylation is required in human disease and animal model studies. We developed a novel antibody-free proteomics assay to measure α-tubulin acetylation targeting protease AspN-generated peptides harboring K40 site. Using the synthetic unmodified and acetylated stable isotope labeled peptides DKTIGGG and DKTIGGGD, we demonstrate assay linearity across 4 log magnitude and reproducibility o

Generation of human-induced pluripotent-stem-cell-derived cortical neurons for high-throughput imaging of neurite morphology and neuron maturation

High-throughput imaging allows in vitro assessment of neuron morphology for screening populations under developmental, homeostatic, and/or disease conditions. Here, we present a protocol to differentiate cryopreserved human cortical neuronal progenitors into mature cortical neurons for high-throughput imaging analysis. We describe the use of a notch signaling inhibitor to generate homogeneous neuronal populations at densities amenable to individual neurite identification. We detail neurite morphology assessment via measuring multiple parameters including neurite length, branches, roots, segments and extremities, and neuron maturation

Women and WASH in Nepal : a scoping review of existing literature

The relationship between gender and water, sanitation and hygiene (WASH) services is an issue for many developing states. This scoping review demonstrates that in Nepal, women bear the major social burden of inadequate WASH services and are under-represented in WASH-related policies and programmes. Four themes emerge: gender and water projects; lack of equal female participation in WASH policy development and implementation; women’s access to water; and menstruation and menstrual hygiene. Major cultural barriers embedded in Nepali society limit women’s access, agency and participation in WASH-related policy and practice

A Patient-Specific Stem Cell Model to Investigate the Neurological Phenotype Observed in Ataxia-Telangiectasia

International audienc

Women and Wash in Nepal: Key Issues and Challenges

This report has been jointly prepared by Western Sydney University’s Humanitarian and Development Research Initiative (HADRI) and Nepal’s National Disaster Risk Reduction Centre (NDRC). This report is thus the result of a strong collaboration between long-term organisational partners: HADRI staff and students have been working in Nepal on humanitarian and development issues — including water, sanitation and hygiene (WASH)— with NDRC and the University of Kathmandu Nepal for several years, and several Nepal-based scholars are Adjunct Fellows of HADRI. The purpose of this report is to provide an overview of the main WASH issues in Nepal with a specific focus on the experiences of women, to set out gaps in knowledge and areas in need of future attention. It offers a stocktake of current research, programmatic interventions and knowledge gaps on WASH in Nepal. The report also highlights challenges for the future. It is based on a comprehensive literature and policy review as well as field assessments drawn from NDRC. The report firstly sets out the broader context in Nepal with regard to social, political, cultural and environmental practices including those that impact on WASH practices. It also records the progress that has been made in many areas of society in spite of significant challenges such as the 2015 earthquake. A literature review summarises existing knowledge in the WASH-related areas of health, hygiene and sanitation, noting the variances due to gender, caste, ethnicity, economic status and location. the report then reviews access to water, paying particular attention to gender, and participation in programme and policy design. A case-study approach illustrates how these WASH-related issues are experienced at the local level in four districts in Nepal, with a policy review setting out the major stages in WASH policy design and implementation since the 1970s. The report concludes by setting out gaps in knowledge and critical areas for future action