35 results on '"Glaß, Hannes"'
Search Results
2. Therapeutic targeting of Lyn kinase to treat chorea-acanthocytosis
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Peikert, Kevin, Federti, Enrica, Matte, Alessandro, Constantin, Gabriela, Pietronigro, Enrica Caterina, Fabene, Paolo Francesco, Defilippi, Paola, Turco, Emilia, Del Gallo, Federico, Pucci, Pietro, Amoresano, Angela, Illiano, Anna, Cozzolino, Flora, Monti, Maria, Garello, Francesca, Terreno, Enzo, Alper, Seth Leo, Glaß, Hannes, Pelzl, Lisann, Akgün, Katja, Ziemssen, Tjalf, Ordemann, Rainer, Lang, Florian, Brunati, Anna Maria, Tibaldi, Elena, Andolfo, Immacolata, Iolascon, Achille, Bertini, Giuseppe, Buffelli, Mario, Zancanaro, Carlo, Lorenzetto, Erika, Siciliano, Angela, Bonifacio, Massimiliano, Danek, Adrian, Walker, Ruth Helen, Hermann, Andreas, and De Franceschi, Lucia
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- 2021
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3. Pathomechanisms of ALS8: altered autophagy and defective RNA binding protein (RBP) homeostasis due to the VAPB P56S mutation
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Tripathi, Priyanka, Guo, Haihong, Dreser, Alice, Yamoah, Alfred, Sechi, Antonio, Jesse, Christopher Marvin, Katona, Istvan, Doukas, Panagiotis, Nikolin, Stefan, Ernst, Sabrina, Aronica, Eleonora, Glaß, Hannes, Hermann, Andreas, Steinbusch, Harry, Feller, Alfred C., Bergmann, Markus, Jaarsma, Dick, Weis, Joachim, and Goswami, Anand
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- 2021
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4. Cell-Type-Dependent Recruitment Dynamics of FUS Protein at Laser-Induced DNA Damage Sites.
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Niu, Yu, Pal, Arun, Szewczyk, Barbara, Japtok, Julia, Naumann, Marcel, Glaß, Hannes, and Hermann, Andreas
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DNA repair ,DNA damage ,AMYOTROPHIC lateral sclerosis ,PLURIPOTENT stem cells ,INDUCED pluripotent stem cells ,SINGLE-stranded DNA - Abstract
Increased signs of DNA damage have been associated to aging and neurodegenerative diseases. DNA damage repair mechanisms are tightly regulated and involve different pathways depending on cell types and proliferative vs. postmitotic states. Amongst them, fused in sarcoma (FUS) was reported to be involved in different pathways of single- and double-strand break repair, including an early recruitment to DNA damage. FUS is a ubiquitously expressed protein, but if mutated, leads to a more or less selective motor neurodegeneration, causing amyotrophic lateral sclerosis (ALS). Of note, ALS-causing mutation leads to impaired DNA damage repair. We thus asked whether FUS recruitment dynamics differ across different cell types putatively contributing to such cell-type-specific vulnerability. For this, we generated engineered human induced pluripotent stem cells carrying wild-type FUS-eGFP and analyzed different derivatives from these, combining a laser micro-irradiation technique and a workflow to analyze the real-time process of FUS at DNA damage sites. All cells showed FUS recruitment to DNA damage sites except for hiPSC, with only 70% of cells recruiting FUS. In-depth analysis of the kinetics of FUS recruitment at DNA damage sites revealed differences among cellular types in response to laser-irradiation-induced DNA damage. Our work suggests a cell-type-dependent recruitment behavior of FUS during the DNA damage response and repair procedure. The presented workflow might be a valuable tool for studying the proteins recruited at the DNA damage site in a real-time course. [ABSTRACT FROM AUTHOR]
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- 2024
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5. Corrosion Products from Metallic Implants Induce ROS and Cell Death in Human Motoneurons In Vitro
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Glaß, Hannes, primary, Jonitz-Heincke, Anika, additional, Petters, Janine, additional, Lukas, Jan, additional, Bader, Rainer, additional, and Hermann, Andreas, additional
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- 2023
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6. Mitochondria–Endoplasmic Reticulum Contact Sites Dynamics and Calcium Homeostasis Are Differentially Disrupted in PINK1‐PD or PRKN‐PD Neurons
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Grossmann, Dajana, primary, Malburg, Nina, additional, Glaß, Hannes, additional, Weeren, Veronika, additional, Sondermann, Verena, additional, Pfeiffer, Julia F., additional, Petters, Janine, additional, Lukas, Jan, additional, Seibler, Philip, additional, Klein, Christine, additional, Grünewald, Anne, additional, and Hermann, Andreas, additional
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- 2023
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7. Restoring Axonal Organelle Motility and Regeneration in Cultured FUS-ALS Motoneurons through Magnetic Field Stimulation Suggests an Alternative Therapeutic Approach
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Kandhavivorn, Wonphorn, primary, Glaß, Hannes, additional, Herrmannsdörfer, Thomas, additional, Böckers, Tobias M., additional, Uhlarz, Marc, additional, Gronemann, Jonas, additional, Funk, Richard H. W., additional, Pietzsch, Jens, additional, Pal, Arun, additional, and Hermann, Andreas, additional
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- 2023
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8. Live Cell Imaging of ATP Levels Reveals Metabolic Compartmentalization within Motoneurons and Early Metabolic Changes in FUS ALS Motoneurons
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Zimyanin, Vitaly L., primary, Pielka, Anna-Maria, additional, Glaß, Hannes, additional, Japtok, Julia, additional, Großmann, Dajana, additional, Martin, Melanie, additional, Deussen, Andreas, additional, Szewczyk, Barbara, additional, Deppmann, Chris, additional, Zunder, Eli, additional, Andersen, Peter M., additional, Boeckers, Tobias M., additional, Sterneckert, Jared, additional, Redemann, Stefanie, additional, Storch, Alexander, additional, and Hermann, Andreas, additional
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- 2023
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9. Live cell imaging of ATP levels reveals metabolic compartmentalization within motoneurons and early metabolic changes inFUSALS motoneurons
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Zimyanin, Vitaly, primary, Pielka, Anne-Marie, additional, Glaß, Hannes, additional, Japtok, Julia, additional, Martin, Melanie, additional, Deussen, Andreas, additional, Szewczyk, Barbara, additional, Deppmann, Chris, additional, Zunder, Eli, additional, Andersen, Peter M., additional, Boeckers, Tobias M., additional, Sterneckert, Jared, additional, Redemann, Stefanie, additional, Storch, Alexander, additional, and Hermann, Andreas, additional
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- 2023
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10. Live cell imaging of ATP levels reveals metabolic compartmentalization within motoneurons and early metabolic changes in FUS ALS motoneurons
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Zimyanin, Vitaly L., Pielka, Anna-Maria, Glaß, Hannes, Japtok, Julia, Großmann, Dajana, Martin, Melanie, Deussen, Andreas, Szewczyk, Barbara, Deppmann, Chris, Zunder, Eli, Andersen, Peter M., Boeckers, Tobias M., Sterneckert, Jared, Redemann, Stefanie, Storch, Alexander, Hermann, Andreas, Zimyanin, Vitaly L., Pielka, Anna-Maria, Glaß, Hannes, Japtok, Julia, Großmann, Dajana, Martin, Melanie, Deussen, Andreas, Szewczyk, Barbara, Deppmann, Chris, Zunder, Eli, Andersen, Peter M., Boeckers, Tobias M., Sterneckert, Jared, Redemann, Stefanie, Storch, Alexander, and Hermann, Andreas
- Abstract
Motoneurons are one of the most energy-demanding cell types and a primary target in Amyotrophic lateral sclerosis (ALS), a debilitating and lethal neurodegenerative disorder without currently available effective treatments. Disruption of mitochondrial ultrastructure, transport, and metabolism is a commonly reported phenotype in ALS models and can critically affect survival and the proper function of motor neurons. However, how changes in metabolic rates contribute to ALS progression is not fully understood yet. Here, we utilize hiPCS-derived motoneuron cultures and live imaging quantitative techniques to evaluate metabolic rates in fused in sarcoma (FUS)-ALS model cells. We show that differentiation and maturation of motoneurons are accompanied by an overall upregulation of mitochondrial components and a significant increase in metabolic rates that correspond to their high energy-demanding state. Detailed compartment-specific live measurements using a fluorescent ATP sensor and FLIM imaging show significantly lower levels of ATP in the somas of cells carrying FUS-ALS mutations. These changes lead to the increased vulnerability of diseased motoneurons to further metabolic challenges with mitochondrial inhibitors and could be due to the disruption of mitochondrial inner membrane integrity and an increase in its proton leakage. Furthermore, our measurements demonstrate heterogeneity between axonal and somatic compartments, with lower relative levels of ATP in axons. Our observations strongly support the hypothesis that mutated FUS impacts the metabolic states of motoneurons and makes them more susceptible to further neurodegenerative mechanisms.
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- 2023
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11. Mitochondria-Endoplasmic Reticulum Contact Sites Dynamics and Calcium Homeostasis Are Differentially Disrupted in PINK1-PD or PRKN-PD Neurons
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Grossmann, Dajana, Malburg, Nina, Glaß, Hannes, Weeren, Veronika, Sondermann, Verena, Pfeiffer, Julia F., Petters, Janine, Lukas, Jan, Seibler, Philip, Klein, Christine, Grünewald, Anne, and Hermann, Andreas
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Biochemistry, biophysics & molecular biology [F05] [Life sciences] ,Biochimie, biophysique & biologie moléculaire [F05] [Sciences du vivant] - Abstract
Background: It is generally believed that the pathogenesis of PINK1/parkin-related Parkinson's disease (PD) is due to a disturbance in mitochondrial quality control. However, recent studies have found that PINK1 and Parkin play a significant role in mitochondrial calcium homeostasis and are involved in the regulation of mitochondria-endoplasmic reticulum contact sites (MERCSs). Objective: The aim of our study was to perform an in-depth analysis of the role of MERCSs and impaired calcium homeostasis in PINK1/Parkin-linked PD.MethodsIn our study, we used induced pluripotent stem cell-derived dopaminergic neurons from patients with PD with loss-of-function mutations in PINK1 or PRKN. We employed a split-GFP-based contact site sensor in combination with the calcium-sensitive dye Rhod-2 AM and applied Airyscan live-cell super-resolution microscopy to determine how MERCSs are involved in the regulation of mitochondrial calcium homeostasis. Results: Our results showed that thapsigargin-induced calcium stress leads to an increase of the abundance of narrow MERCSs in wild-type neurons. Intriguingly, calcium levels at the MERCSs remained stable, whereas the increased net calcium influx resulted in elevated mitochondrial calcium levels. However, PINK1-PD or PRKN-PD neurons showed an increased abundance of MERCSs at baseline, accompanied by an inability to further increase MERCSs upon thapsigargin-induced calcium stress. Consequently, calcium distribution at MERCSs and within mitochondria was disrupted. Conclusions: Our results demonstrated how the endoplasmic reticulum and mitochondria work together to cope with calcium stress in wild-type neurons. In addition, our results suggests that PRKN deficiency affects the dynamics and composition of MERCSs differently from PINK1 deficiency, resulting in differentially affected calcium homeostasis. © 2023 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.
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- 2023
12. Changes in Blood Cell Deformability in Chorea-Acanthocytosis and Effects of Treatment With Dasatinib or Lithium
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Reichel, Felix, primary, Kräter, Martin, additional, Peikert, Kevin, additional, Glaß, Hannes, additional, Rosendahl, Philipp, additional, Herbig, Maik, additional, Rivera Prieto, Alejandro, additional, Kihm, Alexander, additional, Bosman, Giel, additional, Kaestner, Lars, additional, Hermann, Andreas, additional, and Guck, Jochen, additional
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- 2022
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13. Applied Bayesian Approaches for Research in Motor Neuron Disease
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Temp, Anna G. M., primary, Naumann, Marcel, additional, Hermann, Andreas, additional, and Glaß, Hannes, additional
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- 2022
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14. Changes in blood cell deformability in Chorea-Acanthocytosis and effects of treatment with dasatinib or lithium
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Reichel, Felix, primary, Kraeter, Martin, additional, Peikert, Kevin, additional, Glass, Hannes, additional, Rosendahl, Philipp, additional, Herbig, Maik, additional, Prieto, Alejandro Rivera, additional, Kihm, Alexander, additional, Kaestner, Lars, additional, Hermann, Andreas, additional, and Guck, Jochen, additional
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- 2021
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15. The Erythrocyte Sedimentation Rate and Its Relation to Cell Shape and Rigidity of Red Blood Cells from Chorea-Acanthocytosis Patients in an Off-Label Treatment with Dasatinib
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Rabe, Antonia, Kihm, Alexander, Darras, Alexis, Peikert, Kevin, Simionato, Greta, Dasanna, Anil Kumar, Glaß, Hannes, Geisel, Jürgen, Quint, Stephan, Danek, Adrian, Wagner, Christian, Fedosov, Dmitry A., Hermann, Andreas, and Kaestner, Lars
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Adult ,Male ,Erythrocytes ,microfluidics ,therapeutic use [Dasatinib] ,3D shapes ,Blood Sedimentation ,drug effects [Acanthocytes] ,cell rigidity ,Microbiology ,Article ,ddc:570 ,therapeutic use [Protein Kinase Inhibitors] ,Humans ,dasatinib ,acanthocytes ,Cell Shape ,Protein Kinase Inhibitors ,drug effects [Cell Shape] ,blood [Biomarkers] ,drug therapy [Neuroacanthocytosis] ,Off-Label Use ,QR1-502 ,pathology [Acanthocytes] ,phase diagram ,mesoscopic modeling ,drug effects [Blood Sedimentation] ,drug effects [Erythrocytes] ,erythrocyte sedimentation rate ,blood [Neuroacanthocytosis] ,Biomarkers ,Neuroacanthocytosis ,pathology [Neuroacanthocytosis] - Abstract
Background: Chorea-acanthocytosis (ChAc) is a rare hereditary neurodegenerative disease with deformed red blood cells (RBCs), so-called acanthocytes, as a typical marker of the disease. Erythrocyte sedimentation rate (ESR) was recently proposed as a diagnostic biomarker. To date, there is no treatment option for affected patients, but promising therapy candidates, such as dasatinib, a Lyn-kinase inhibitor, have been identified. Methods: RBCs of two ChAc patients during and after dasatinib treatment were characterized by the ESR, clinical hematology parameters and the 3D shape classification in stasis based on an artificial neural network. Furthermore, mathematical modeling was performed to understand the contribution of cell morphology and cell rigidity to the ESR. Microfluidic measurements were used to compare the RBC rigidity between ChAc patients and healthy controls. Results: The mechano-morphological characterization of RBCs from two ChAc patients in an off-label treatment with dasatinib revealed differences in the ESR and the acanthocyte count during and after the treatment period, which could not directly be related to each other. Clinical hematology parameters were in the normal range. Mathematical modeling indicated that RBC rigidity is more important for delayed ESR than cell shape. Microfluidic experiments confirmed a higher rigidity in the normocytes of ChAc patients compared to healthy controls. Conclusions: The results increase our understanding of the role of acanthocytes and their associated properties in the ESR, but the data are too sparse to answer the question of whether the ESR is a suitable biomarker for treatment success, whereas a correlation between hematological and neuronal phenotype is still subject to verification.
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- 2021
16. Reduced Expression of GABAA Receptor Alpha2 Subunit Is Associated With Disinhibition of DYT-THAP1 Dystonia Patient-Derived Striatal Medium Spiny Neurons
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Staege, Selma, primary, Kutschenko, Anna, additional, Baumann, Hauke, additional, Glaß, Hannes, additional, Henkel, Lisa, additional, Gschwendtberger, Thomas, additional, Kalmbach, Norman, additional, Klietz, Martin, additional, Hermann, Andreas, additional, Lohmann, Katja, additional, Seibler, Philip, additional, and Wegner, Florian, additional
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- 2021
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17. Targeting Lyn Kinase in Chorea-Acanthocytosis: A Translational Treatment Approach in a Rare Disease
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Peikert, Kevin, Glaß, Hannes, Federti, Enrica, Matte, Alessandro, Pelzl, Lisann, Akgün, Katja, Ziemssen, Tjalf, Ordemann, Rainer, Lang, Florian, Patients, The Network for Translational Research for Neuroacanthocytosis, Franceschi, Lucia De, and Hermann, Andreas
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off-label ,ChAc ,Medicine ,dasatinib ,ddc:610 ,neuroacanthocytosis ,TKI ,Article - Abstract
Chorea-acanthocytosis (ChAc) is a neurodegenerative disease caused by mutations in the VPS13A gene. It is characterized by several neurological symptoms and the appearance of acanthocytes. Elevated tyrosine kinase Lyn activity has been recently identified as one of the key pathophysiological mechanisms in this disease, and therefore represents a promising drug target. Methods: We evaluated an individual off-label treatment with the tyrosine kinase inhibitor dasatinib (100 mg/d, 25.8–50.4 weeks) of three ChAc patients. Alongside thorough safety monitoring, we assessed motor and non-motor scales (e.g., MDS-UPDRS, UHDRS, quality of life) as well as routine and experimental laboratory parameters (e.g., serum neurofilament, Lyn kinase activity, actin cytoskeleton in red blood cells). Results: Dasatinib appeared to be reasonably safe. The clinical parameters remained stable without significant improvement or deterioration. Regain of deep tendon reflexes was observed in one patient. Creatine kinase, serum neurofilament levels, and acanthocyte count did not reveal consistent effects. However, a reduction of initially elevated Lyn kinase activity and accumulated autophagy markers, as well as a partial restoration of the actin cytoskeleton, was found in red blood cells. Conclusions: We report on the first treatment approach with disease-modifying intention in ChAc. The experimental parameters indicate target engagement in red blood cells, while clinical effects on the central nervous system could not be proven within a rather short treatment time. Limited knowledge on the natural history of ChAc and the lack of appropriate biomarkers remain major barriers for “clinical trial readiness”. We suggest a panel of outcome parameters for future clinical trials in ChAc.
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- 2021
18. A serum microRNA sequence reveals fragile X protein pathology in amyotrophic lateral sclerosis
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Freischmidt, Axel, Goswami, Anand, Limm, Katharina, Zimyanin, Vitaly L., Demestre, Maria, Glaß, Hannes, Holzmann, Karlheinz, Helferich, Anika M., Brockmann, Sarah J., Tripathi, Priyanka, Yamoah, Alfred, Poser, Ina, Oefner, Peter J., Böckers, Tobias M., Aronica, Eleonora, Ludolph, Albert C., Andersen, Peter M., Hermann, Andreas, Weis, Joachim, Reinders, Jörg, Danzer, Karin M., Weishaupt, Jochen H., Freischmidt, Axel, Goswami, Anand, Limm, Katharina, Zimyanin, Vitaly L., Demestre, Maria, Glaß, Hannes, Holzmann, Karlheinz, Helferich, Anika M., Brockmann, Sarah J., Tripathi, Priyanka, Yamoah, Alfred, Poser, Ina, Oefner, Peter J., Böckers, Tobias M., Aronica, Eleonora, Ludolph, Albert C., Andersen, Peter M., Hermann, Andreas, Weis, Joachim, Reinders, Jörg, Danzer, Karin M., and Weishaupt, Jochen H.
- Abstract
Knowledge about converging disease mechanisms in the heterogeneous syndrome amyotrophic lateral sclerosis (ALS) is rare, but may lead to therapies effective in most ALS cases. Previously, we identified serum microRNAs downregulated in familial ALS, the majority of sporadic ALS patients, but also in presymptomatic mutation carriers. A 5-nucleotide sequence motif (GDCGG; D = G, A or U) was strongly enriched in these ALS-related microRNAs. We hypothesized that deregulation of protein(s) binding predominantly to this consensus motif was responsible for the ALS-linked microRNA fingerprint. Using microRNA pull-down assays combined with mass spectrometry followed by extensive biochemical validation, all members of the fragile X protein family, FMR1, FXR1 and FXR2, were identified to directly and predominantly interact with GDCGG microRNAs through their structurally disordered RGG/RG domains. Preferential association of this protein family with ALS-related microRNAs was confirmed by in vitro binding studies on a transcriptome-wide scale. Immunohistochemistry of lumbar spinal cord revealed aberrant expression level and aggregation of FXR1 and FXR2 in C9orf72- and FUS-linked familial ALS, but also patients with sporadic ALS. Further analysis of ALS autopsies and induced pluripotent stem cell-derived motor neurons with FUS mutations showed co-aggregation of FXR1 with FUS. Hence, our translational approach was able to take advantage of blood microRNAs to reveal CNS pathology, and suggests an involvement of the fragile X-related proteins in familial and sporadic ALS already at a presymptomatic stage. The findings may uncover disease mechanisms relevant to many patients with ALS. They furthermore underscore the systemic, extra-CNS aspect of ALS.
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- 2021
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19. Pathomechanisms of ALS8:altered autophagy and defective RNA binding protein (RBP) homeostasis due to the VAPB P56S mutation
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Tripathi, Priyanka, Guo, Haihong, Dreser, Alice, Yamoah, Alfred, Sechi, Antonio, Jesse, Christopher Marvin, Katona, Istvan, Doukas, Panagiotis, Nikolin, Stefan, Ernst, Sabrina, Aronica, Eleonora, Glaß, Hannes, Hermann, Andreas, Steinbusch, Harry, Feller, Alfred C., Bergmann, Markus, Jaarsma, Dick, Weis, Joachim, Goswami, Anand, Tripathi, Priyanka, Guo, Haihong, Dreser, Alice, Yamoah, Alfred, Sechi, Antonio, Jesse, Christopher Marvin, Katona, Istvan, Doukas, Panagiotis, Nikolin, Stefan, Ernst, Sabrina, Aronica, Eleonora, Glaß, Hannes, Hermann, Andreas, Steinbusch, Harry, Feller, Alfred C., Bergmann, Markus, Jaarsma, Dick, Weis, Joachim, and Goswami, Anand
- Abstract
Mutations in RNA binding proteins (RBPs) and in genes regulating autophagy are frequent causes of familial amyotrophic lateral sclerosis (fALS). The P56S mutation in vesicle-associated membrane protein-associated protein B (VAPB) leads to fALS (ALS8) and spinal muscular atrophy (SMA). While VAPB is primarily involved in the unfolded protein response (UPR), vesicular trafficking and in initial steps of the autophagy pathway, the effect of mutant P56S-VAPB on autophagy regulation in connection with RBP homeostasis has not been explored yet. Examining the muscle biopsy of our index ALS8 patient of European origin revealed globular accumulations of VAPB aggregates co-localised with autophagy markers LC3 and p62 in partially atrophic and atrophic muscle fibres. In line with this skin fibroblasts obtained from the same patient showed accumulation of P56S-VAPB aggregates together with LC3 and p62. Detailed investigations of autophagic flux in cell culture models revealed that P56S-VAPB alters both initial and late steps of the autophagy pathway. Accordingly, electron microscopy complemented with live cell imaging highlighted the impaired fusion of accumulated autophagosomes with lysosomes in cells expressing P56S-VAPB. Consistent with these observations, neuropathological studies of brain and spinal cord of P56S-VAPB transgenic mice revealed signs of neurodegeneration associated with altered protein quality control and defective autophagy. Autophagy and RBP homeostasis are interdependent, as demonstrated by the cytoplasmic mis-localisation of several RBPs including pTDP-43, FUS, Matrin 3 which often sequestered with P56S-VAPB aggregates both in cell culture and in the muscle biopsy of the ALS8 patient. Further confirming the notion that aggregation of the RBPs proceeds through the stress granule (SG) pathway, we found persistent G3BP- and TIAR1-positive SGs in P56S-VAPB expressing cells as well as in the ALS8 patient muscle biopsy. We conclude that P56S-VAPB-ALS8 involv
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- 2021
20. A serum microRNA sequence reveals fragile X protein pathology in amyotrophic lateral sclerosis
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Freischmidt, Axel, primary, Goswami, Anand, additional, Limm, Katharina, additional, Zimyanin, Vitaly L, additional, Demestre, Maria, additional, Glaß, Hannes, additional, Holzmann, Karlheinz, additional, Helferich, Anika M, additional, Brockmann, Sarah J, additional, Tripathi, Priyanka, additional, Yamoah, Alfred, additional, Poser, Ina, additional, Oefner, Peter J, additional, Böckers, Tobias M, additional, Aronica, Eleonora, additional, Ludolph, Albert C, additional, Andersen, Peter M, additional, Hermann, Andreas, additional, Weis, Joachim, additional, Reinders, Jörg, additional, Danzer, Karin M, additional, and Weishaupt, Jochen H, additional
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- 2021
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21. Functional and Molecular Properties of DYT-SGCE Myoclonus-Dystonia Patient-Derived Striatal Medium Spiny Neurons
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Kutschenko, Anna, primary, Staege, Selma, additional, Grütz, Karen, additional, Glaß, Hannes, additional, Kalmbach, Norman, additional, Gschwendtberger, Thomas, additional, Henkel, Lisa M., additional, Heine, Johanne, additional, Grünewald, Anne, additional, Hermann, Andreas, additional, Seibler, Philip, additional, and Wegner, Florian, additional
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- 2021
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22. Human Spinal Motor Neurons Are Particularly Vulnerable to Cerebrospinal Fluid of Amyotrophic Lateral Sclerosis Patients
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Bräuer, Stefan, primary, Günther, René, additional, Sterneckert, Jared, additional, Glaß, Hannes, additional, and Hermann, Andreas, additional
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- 2020
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23. High content live profiling reveals concomitant gain and loss of function pathomechanisms in C9ORF72 amyotrophic lateral sclerosis
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Pal, Arun, primary, Kretner, Benedikt, additional, Abo-Rady, Masin, additional, Glaß, Hannes, additional, Naumann, Marcel, additional, Japtok, Julia, additional, Kreiter, Nicole, additional, Böckers, Tobias M., additional, Sterneckert, Jared, additional, and Hermann, Andreas, additional
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- 2020
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24. Combined Dendritic and Axonal Deterioration Are Responsible for Motoneuronopathy in Patient-Derived Neuronal Cell Models of Chorea-Acanthocytosis
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Glaß, Hannes, primary, Neumann, Patrick, additional, Pal, Arun, additional, Reinhardt, Peter, additional, Storch, Alexander, additional, Sterneckert, Jared, additional, and Hermann, Andreas, additional
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- 2020
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25. Knocking out C9ORF72 Exacerbates Axonal Trafficking Defects Associated with Hexanucleotide Repeat Expansion and Reduces Levels of Heat Shock Proteins
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Abo-Rady, Masin, primary, Kalmbach, Norman, additional, Pal, Arun, additional, Schludi, Carina, additional, Janosch, Antje, additional, Richter, Tanja, additional, Freitag, Petra, additional, Bickle, Marc, additional, Kahlert, Anne-Karin, additional, Petri, Susanne, additional, Stefanov, Stefan, additional, Glass, Hannes, additional, Staege, Selma, additional, Just, Walter, additional, Bhatnagar, Rajat, additional, Edbauer, Dieter, additional, Hermann, Andreas, additional, Wegner, Florian, additional, and Sterneckert, Jared L., additional
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- 2020
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26. High content organelle trafficking enables disease state profiling as powerful tool for disease modelling
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Pal, Arun, primary, Glaß, Hannes, additional, Naumann, Marcel, additional, Kreiter, Nicole, additional, Japtok, Julia, additional, Sczech, Ronny, additional, and Hermann, Andreas, additional
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- 2018
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27. Cell culture models of Chorea Acanthocytosis and their evaluation
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Hermann, Andreas, Tanaka, Elly, Technische Universität Dresden, Glaß, Hannes, Hermann, Andreas, Tanaka, Elly, Technische Universität Dresden, and Glaß, Hannes
- Abstract
Chorea Acanthocytosis (ChAc) is an autosomal recessive inherited disease caused by loss- of-function mutation in the VPS13A gene which encodes CHOREIN protein. This study used induced pluripotent stem cells (iPSCs) as well as neural progenitor cells (NPCs) to generate medium spiny neurons (MSN) as well as midbrain dopaminergic neurons (mDAN). The first objective of this thesis was to generate and characterize a stem cell based disease model of ChAc. The second objective was to establish two different differentiation protocols that yield different neuronal sub types that are affected in ChAc, and compare whether they harbor similar phenotypes and whether the faster protocol can be used to model the disease accurately. The generated iPSCs were characterized using AP staining as an early marker for reprogramming, qPCR for analysis of residual expression of exogenous transcription factors, immunocytochemistry (ICC) for staining of pluripotency markers as well as markers for mesoderm, ectoderm and endoderm formation upon three germ layer formation. Karyotyping was conducted to exclude aberrant clones. Western blot using CHOREIN antibody revealed that the cell lines retained their disease identity. There were no differences observed between wild type and ChAc lines in stem cell and neuron populations in either protocol. qPCR analysis, investigating the expression of previously described markers for characterization, revealed no significant clustering between wild type and ChAc lines in either protocol. A disturbed ratio of globular and filamentous actin is causative for the aberrant shape of ChAc erythrocytes. Investigation of the ratio in mature neurons revealed a significant reduction of this ratio in MSN but no difference in mDAN cultures. When the ratio of cytosolic and filamentous tubulin and the acetylation of tubulin were investigated, no differences were found between wild type and ChAc lines. Mature neurons of both differentiation protocols were subjected to trea
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- 2018
28. Cell culture models of Chorea Acanthocytosis and their evaluation
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Glaß, Hannes, Hermann, Andreas, Tanaka, Elly, and Technische Universität Dresden
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Basic research, Lifescience, Disease modelling ,ddc:570 ,Biomedizin ,Neurologie ,Choreoakanthozytose ,Grundlagenforschung, Biomedizin, Krankheitsmodellierung - Abstract
Chorea Acanthocytosis (ChAc) is an autosomal recessive inherited disease caused by loss- of-function mutation in the VPS13A gene which encodes CHOREIN protein. This study used induced pluripotent stem cells (iPSCs) as well as neural progenitor cells (NPCs) to generate medium spiny neurons (MSN) as well as midbrain dopaminergic neurons (mDAN). The first objective of this thesis was to generate and characterize a stem cell based disease model of ChAc. The second objective was to establish two different differentiation protocols that yield different neuronal sub types that are affected in ChAc, and compare whether they harbor similar phenotypes and whether the faster protocol can be used to model the disease accurately. The generated iPSCs were characterized using AP staining as an early marker for reprogramming, qPCR for analysis of residual expression of exogenous transcription factors, immunocytochemistry (ICC) for staining of pluripotency markers as well as markers for mesoderm, ectoderm and endoderm formation upon three germ layer formation. Karyotyping was conducted to exclude aberrant clones. Western blot using CHOREIN antibody revealed that the cell lines retained their disease identity. There were no differences observed between wild type and ChAc lines in stem cell and neuron populations in either protocol. qPCR analysis, investigating the expression of previously described markers for characterization, revealed no significant clustering between wild type and ChAc lines in either protocol. A disturbed ratio of globular and filamentous actin is causative for the aberrant shape of ChAc erythrocytes. Investigation of the ratio in mature neurons revealed a significant reduction of this ratio in MSN but no difference in mDAN cultures. When the ratio of cytosolic and filamentous tubulin and the acetylation of tubulin were investigated, no differences were found between wild type and ChAc lines. Mature neurons of both differentiation protocols were subjected to treatment with the proteotoxic stress inducer L-canavanine and the unfolded protein response (UPR) inducer tunicamycin. Survival was analyzed with the PrestoBlue assay as well as lactate dehydroxylase (LDH) release assay. Both cultures of mature neurons showed an increased susceptibility to the respective drugs. Furthermore the data suggests that MSN cultures are more vulnerable against proteotoxic stress (L-canavanine). Kinetics of tunicamycin poisoning were not different within MSN cultures but indicated a late cell death of ChAc lines under mDAN differentiation conditions. DNA damage plays a major role in the progression of neurodegenerative diseases. The amount of double strand breaks (DSB) was assessed in mature cultures of MSN and mDAN differentiations. There was no difference in basal level of DSB. When etoposide was applied to induce DNA damage, increased susceptibility of ChAc lines was observed. Albeit significant, the effect size was very small. Seahorse was used to characterize energy metabolism. Glycolysis was not impaired in ChAc lines in either protocol. Furthermore, MSN differentiation showed no difference in any parameter related to oxidative phosphorylation, while under mDAN conditions, coupling efficiency and spare respiratory capacity was increased for ChAc lines. The non-respiratory oxygen consumption was increased in ChAc lines in MSN cultures but decreased in mDAN cultures. The yeast homolog of VPS13A interacts with vesicle and mitochondrial membranes. Therefore, this study focuses on vesicle and mitochondria homeostasis. Live cell imaging of mature neurons of MSN differentiations revealed a decreased amount and reduced motility of mitochondria. Even though mitochondria were normally shaped their size was reduced. mDAN differentiations harbored a reduced amount and shortened mitochondria. These mitochondria, however, showed an increased motility. When analyzing aligned mature neurons in microfluidic chambers (MFCs), a strong phenotype was already observed in proximal regions, which resembled the distal parts of the channels. Hence, the dysregulation, that occurs distal in healthy controls, happens closer to the soma in diseased cells. The mitochondria potential marker JC-1 showed a hyperpolarization of mitochondria in MSN culture and a depolarization in mDAN cultures. When investigated in MFCs of mDAN cultures, there was a significant increase in potential observed at the distal position of ChAc lines, while wild type cultures showed no difference. Experiments conducted on the lysosomal compartments showed a decrease in proximal parts of ChAc MSN cultures when compared to wild type. Their shape was altered as well. mDAN cultures featured no significant morphological changes. Trafficking analysis revealed an increase in motility in MSN cultures but a decrease in mDAN cultures. When lysosomes were analyzed in MFCs only mDAN cultures showed an increase in retrograde transport. In order to investigate whether the in vitro phenotypes of Huntington (Htt) and ChAc are similar, some of the previous experiments were conducted in MSN differentiations of one Htt line. Cells from Htt behaved similar to ChAc lines when DNA damage response was investigated. Analysis of mitochondrial parameters showed no difference as well. However, the non-respiratory oxygen consumption was not increased and resembled wild type. When Htt neurons were investigated during live cell imaging, shortened mitochondria were found. Their number was not reduced significantly. However, a trend for reduction was observed. Mitochondria of Htt cells were more motile than ChAc or wild type lines. Mitochondrial potential was increased in Htt and comparable to ChAc. Lysosomal count showed a reduction and the area of Htt lysosomes was significantly smaller than wild type or ChAc. Lysosomes of Htt cells were more motile than their wild type or ChAc counterparts.:List of abbreviations Introduction 1. Neurodegenerative diseases 1.1. Chorea-acanthocytosis – a clinical overview 1.2. Chorea-Acanthocytosis – genetic considerations 2. Disease modelling 2.1. Human disease models 2.2. Induced pluripotent stem cells 2.3. Multipotent neuronal progenitor cells 3. Objectives of this thesis Materials & Methods 1. Cell culture procedures 1.1. Coating 1.2. Matrigel 1.3. PLO/laminin 1.4. Gelatin coating 1.5. Mouse embryonic fibroblast isolation 1.6. Generation of feeder cells 1.7. Human fibroblast culture 1.8. Reprogramming 1.9. iPSC culture 1.10. Culture of small molecule neuronal precursor cells (smNPC) 1.11. MSN differentiation 1.12. mDAN differentiation 2. Nucleic acid biochemistry 2.1. mRNA isolation 2.2. cDNA generation 2.3. Polymerase chain reaction (PCR) 2.4. Agarose gel electrophoresis 3. Cell survival analysis 3.1. PrestoBlue cell viability assay 3.2. Cytotoxicity detection kit: 3.3. DNA damage analysis 4. Metabolic characterization 5. Protein biochemistry 5.1. Alkaline phosphatase staining 5.2. Preparation of immunocytochemistry samples 5.3. Isolation of globular and filamentous actin 5.4. Whole cell protein Isolation 5.5. Cytosolic protein isolation 5.6. Protein concentration measurement 5.7. Western blot 6. Live cell imaging 7. Statistics Results 1. Generation of induced pluripotent stem cells 1.1. Silencing of exogenous transcription factors 1.2. Karyotyping of iPSC clones 1.3. Evaluation of pluripotency 1.4. Alkaline phosphatase staining 1.5. Staining of pluripotency markers 1.6. Three germ layer formation 1.7. Confirmation of ChAc phenotype by CHOREIN western blot 2. Characterization of differentiation potential 2.1. Differentiation efficiency 2.2. Characterization by qPCR 2.3. Ratio of polymerized and unpolymerized cytoskeleton proteins 2.4. Cell survival upon stress induction 2.5. DNA damage in mature neurons 2.6. Characterization of metabolism 3. Live cell imaging 3.1. Mitochondrial dynamics 3.1.1. Morphological analysis 3.1.1.1. Undirected neurons (96 well plate format) 3.1.1.2. Microfluidic chambers 3.1.2. Trafficking analysis 3.1.2.1. 96 well 3.1.2.2. Microfluidic chambers 3.1.3. JC-1 3.1.3.1. 96 well 3.1.3.2. Microfluidic chambers 3.2. Lysosomal dynamics 3.2.1. Morphological analysis 3.2.1.1. 96 well 3.2.1.2. Microfluidic chambers 3.2.2. Trafficking 3.2.2.1. 96 well 3.2.2.2. Microfluidic chambers 4. Comparison with Huntington’s disease 4.1. DNA damage 4.2. Characterization of metabolism 4.3. Live cell imaging 4.3.1. Mitochondria 4.3.1.1. Morphological analysis 4.3.1.2. Trafficking 4.3.1.3. JC-1 4.3.2. Lysosomes 4.3.2.1. Morphological analysis 4.3.2.2. Trafficking Discussion 1. Characterization of ChAc lines 1.1. ChAc stem cell lines show no impaired differentiation potential 1.2. Neurons from MSN differentiation have an altered G/F actin ratio 1.3. Mature neurons from ChAc lines are susceptible to UPR, proteotoxicity and DNA damage 1.4. ChAc neurons are not susceptible to DNA damage 1.5. Energy dynamics in ChAc and Huntington lines feature a shift to glycolysis 2. Live cell imaging of ChAc lines 2.1. Video analysis is reproducible and sensitive 2.2. ChAc lines have altered mitochondria shape and trafficking 2.3. Treatments are not selective on ChAc lines mitochondria 2.4. Mitochondrial potential is altered in ChAc lines 2.5. ChAc lysosomes feature normal morphology but altered trafficking 2.6. Lysosomes of MSN cultures respond poorly to treatments 3. MSN and mDAN differentiation highlight different aspects of the disease References List of figures List of tables Acknowledgments Appendix
- Published
- 2017
29. Defective mitochondrial and lysosomal trafficking in chorea-acanthocytosis is independent of Src-kinase signaling
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Glaß, Hannes, primary, Pal, Arun, additional, Reinhardt, Peter, additional, Sterneckert, Jared, additional, Wegner, Florian, additional, Storch, Alexander, additional, and Hermann, Andreas, additional
- Published
- 2018
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30. Neuronal Dysfunction in iPSC-Derived Medium Spiny Neurons from Chorea-Acanthocytosis Patients Is Reversed by Src Kinase Inhibition and F-Actin Stabilization
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Stanslowsky, Nancy, primary, Reinhardt, Peter, additional, Glass, Hannes, additional, Kalmbach, Norman, additional, Naujock, Maximilian, additional, Hensel, Niko, additional, Lübben, Verena, additional, Pal, Arun, additional, Venneri, Anna, additional, Lupo, Francesca, additional, De Franceschi, Lucia, additional, Claus, Peter, additional, Sterneckert, Jared, additional, Storch, Alexander, additional, Hermann, Andreas, additional, and Wegner, Florian, additional
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- 2016
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31. Differentiation Efficiency of Induced Pluripotent Stem Cells Depends on the Number of Reprogramming Factors
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Löhle, Matthias, primary, Hermann, Andreas, additional, Glaß, Hannes, additional, Kempe, Andrea, additional, Schwarz, Sigrid C., additional, Kim, Jeong Beom, additional, Poulet, Claire, additional, Ravens, Ursula, additional, Schwarz, Johannes, additional, Schöler, Hans R., additional, and Storch, Alexander, additional
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- 2012
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32. Additional file 1 of Therapeutic targeting of Lyn kinase to treat chorea-acanthocytosis
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Peikert, Kevin, Federti, Enrica, Matte, Alessandro, Constantin, Gabriela, Pietronigro, Enrica Caterina, Fabene, Paolo Francesco, Defilippi, Paola, Turco, Emilia, Gallo, Federico Del, Pucci, Pietro, Amoresano, Angela, Illiano, Anna, Cozzolino, Flora, Monti, Maria, Garello, Francesca, Terreno, Enzo, Alper, Seth Leo, Glaß, Hannes, Pelzl, Lisann, Akgün, Katja, Tjalf Ziemssen, Ordemann, Rainer, Lang, Florian, Brunati, Anna Maria, Tibaldi, Elena, Andolfo, Immacolata, Iolascon, Achille, Bertini, Giuseppe, Buffelli, Mario, Zancanaro, Carlo, Lorenzetto, Erika, Siciliano, Angela, Bonifacio, Massimiliano, Danek, Adrian, Walker, Ruth Helen, Hermann, Andreas, and De Franceschi, Lucia
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Data_FILES ,3. Good health - Abstract
Additional file 1.
33. Additional file 1 of Therapeutic targeting of Lyn kinase to treat chorea-acanthocytosis
- Author
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Peikert, Kevin, Federti, Enrica, Matte, Alessandro, Constantin, Gabriela, Pietronigro, Enrica Caterina, Fabene, Paolo Francesco, Defilippi, Paola, Turco, Emilia, Gallo, Federico Del, Pucci, Pietro, Amoresano, Angela, Illiano, Anna, Cozzolino, Flora, Monti, Maria, Garello, Francesca, Terreno, Enzo, Alper, Seth Leo, Glaß, Hannes, Pelzl, Lisann, Akgün, Katja, Tjalf Ziemssen, Ordemann, Rainer, Lang, Florian, Brunati, Anna Maria, Tibaldi, Elena, Andolfo, Immacolata, Iolascon, Achille, Bertini, Giuseppe, Buffelli, Mario, Zancanaro, Carlo, Lorenzetto, Erika, Siciliano, Angela, Bonifacio, Massimiliano, Danek, Adrian, Walker, Ruth Helen, Hermann, Andreas, and De Franceschi, Lucia
- Subjects
Data_FILES ,3. Good health - Abstract
Additional file 1.
34. Reduced Expression of GABA A Receptor Alpha2 Subunit Is Associated With Disinhibition of DYT-THAP1 Dystonia Patient-Derived Striatal Medium Spiny Neurons.
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Staege S, Kutschenko A, Baumann H, Glaß H, Henkel L, Gschwendtberger T, Kalmbach N, Klietz M, Hermann A, Lohmann K, Seibler P, and Wegner F
- Abstract
DYT-THAP1 dystonia (formerly DYT6) is an adolescent-onset dystonia characterized by involuntary muscle contractions usually involving the upper body. It is caused by mutations in the gene THAP1 encoding for the transcription factor Thanatos-associated protein (THAP) domain containing apoptosis-associated protein 1 and inherited in an autosomal-dominant manner with reduced penetrance. Alterations in the development of striatal neuronal projections and synaptic function are known from transgenic mice models. To investigate pathogenetic mechanisms, human induced pluripotent stem cell (iPSC)-derived medium spiny neurons (MSNs) from two patients and one family member with reduced penetrance carrying a mutation in the gene THAP1 (c.474delA and c.38G > A) were functionally characterized in comparison to healthy controls. Calcium imaging and quantitative PCR analysis revealed significantly lower Ca
2+ amplitudes upon GABA applications and a marked downregulation of the gene encoding the GABAA receptor alpha2 subunit in THAP1 MSNs indicating a decreased GABAergic transmission. Whole-cell patch-clamp recordings showed a significantly lower frequency of miniature postsynaptic currents (mPSCs), whereas the frequency of spontaneous action potentials (APs) was elevated in THAP1 MSNs suggesting that decreased synaptic activity might have resulted in enhanced generation of APs. Our molecular and functional data indicate that a reduced expression of GABAA receptor alpha2 subunit could eventually lead to limited GABAergic synaptic transmission, neuronal disinhibition, and hyperexcitability of THAP1 MSNs. These data give pathophysiological insight and may contribute to the development of novel treatment strategies for DYT-THAP1 dystonia., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Staege, Kutschenko, Baumann, Glaß, Henkel, Gschwendtberger, Kalmbach, Klietz, Hermann, Lohmann, Seibler and Wegner.)- Published
- 2021
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35. Targeting Lyn Kinase in Chorea-Acanthocytosis: A Translational Treatment Approach in a Rare Disease.
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Peikert K, Glaß H, Federti E, Matte A, Pelzl L, Akgün K, Ziemssen T, Ordemann R, Lang F, Patients TNFTRFN, De Franceschi L, and Hermann A
- Abstract
Chorea-acanthocytosis (ChAc) is a neurodegenerative disease caused by mutations in the VPS13A gene. It is characterized by several neurological symptoms and the appearance of acanthocytes. Elevated tyrosine kinase Lyn activity has been recently identified as one of the key pathophysiological mechanisms in this disease, and therefore represents a promising drug target. Methods: We evaluated an individual off-label treatment with the tyrosine kinase inhibitor dasatinib (100 mg/d, 25.8-50.4 weeks) of three ChAc patients. Alongside thorough safety monitoring, we assessed motor and non-motor scales (e.g., MDS-UPDRS, UHDRS, quality of life) as well as routine and experimental laboratory parameters (e.g., serum neurofilament, Lyn kinase activity, actin cytoskeleton in red blood cells). Results: Dasatinib appeared to be reasonably safe. The clinical parameters remained stable without significant improvement or deterioration. Regain of deep tendon reflexes was observed in one patient. Creatine kinase, serum neurofilament levels, and acanthocyte count did not reveal consistent effects. However, a reduction of initially elevated Lyn kinase activity and accumulated autophagy markers, as well as a partial restoration of the actin cytoskeleton, was found in red blood cells. Conclusions: We report on the first treatment approach with disease-modifying intention in ChAc. The experimental parameters indicate target engagement in red blood cells, while clinical effects on the central nervous system could not be proven within a rather short treatment time. Limited knowledge on the natural history of ChAc and the lack of appropriate biomarkers remain major barriers for "clinical trial readiness". We suggest a panel of outcome parameters for future clinical trials in ChAc.
- Published
- 2021
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