scholarly journals Norepinephrine transporter defects lead to sympathetic hyperactivity in stem cell and mouse models of Familial Dysautonomia

2021 ◽  
Author(s):  
Hsueh Fu Wu ◽  
Wenxin Yu ◽  
Joeseph Carey ◽  
Frances Lefcort ◽  
Hongxiang Liu ◽  
...  
Keyword(s):  
Nervous System ◽  
Stem Cell ◽  
Drug Testing ◽  
Neurodegenerative Disorder ◽  
Sympathetic Neuron ◽  
Familial Dysautonomia ◽  
Target Tissue ◽  
Future Drug ◽  
Screening Platform

Abstract Familial dysautonomia (FD) is a rare neurodevelopmental and neurodegenerative disorder that affects the sympathetic nervous system. Patients harbor a mutation in ELP1 yet, how loss of Elp1 affects the function of symNs remains unresolved. Such an understanding is critical since the most debilitating hallmarks of the disease include cardiovascular instability, dysautonomic crises and renal failure, which all result from dysregulated sympathetic activity. Here, we employ the human pluripotent stem cell (hPSC) technology as a modeling system to understand human, sympathetic neuron (symN)-specific disease mechanisms and provide a platform for drug testing and discovery. We show that FD symNs are intrinsically hyperactive in vitro, in co-cultures with cardiomyocyte target tissue and in FD animal models. We show that ELP1-rescued isogenic lines remain hyperactive, suggesting a different/additional disease mechanism. Accordingly, we report decreased intracellular norepinephrine (NE) levels, decreased NE re-uptake via NET and excessive extracellular NE in FD symNs. Finally, we performed a mini drug screen showing that current and new candidate drugs were able to lower hyperactivity. These findings may have implications for other peripheral nervous system disorders. Our drug screening platform may allow future drug testing and discovery for such disorders.

Predictive Modeling and Biomechanical Microengineering of Mesenchymal Stem Cells: A High Content Screening Platform to Enhance Lineage Specific Differentiation

2013 ◽  
Author(s):  
Amit Paul ◽  
David Franz ◽  
Sumaira Yahya ◽  
Shan Sun ◽  
Michael Cho
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Predictive Modeling ◽  
Stem Cell Differentiation ◽  
High Content Screening ◽  
Cell Behavior ◽  
Functional Changes ◽  
Screening Platform

Recent evidence suggests that stem cell differentiation can be regulated by modulation of the cell’s biomechanics. The cytoskeletal structures and arrangements in stem cells undergoing differentiation are dramatically altered, and these alterations vary by lineage. The complexity of events associated with the transformation of these precursor cells leaves many questions unanswered about morphological, structural, proteomic, and functional changes in differentiating stem cells. A thorough understanding of stem cell behavior, both experimentally and computationally, would allow for the development of more effective approaches to the expansion of stem cells in vitro and for the regulation of their commitment to a specific phenotype.

In Vitro Electrophysiological Drug Testing Using Human Embryonic Stem Cell Derived Cardiomyocytes

2009 ◽  
Vol 18 (1) ◽  
pp. 161-172 ◽  
Author(s):  
Oren Caspi ◽  
Ilanit Itzhaki ◽  
Izhak Kehat ◽  
Amira Gepstein ◽  
Gil Arbel ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Drug Testing ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem

Human stem cell-derived encapsulated liver tissue as an effective, consistent and long-lasting in vitro tool for drug testing and development

2017 ◽  
Vol 49 (4) ◽  
pp. e251
Author(s):  
C. Raggi ◽  
M. M’Callum ◽  
C. Mangahas ◽  
Z. Cohen ◽  
A. Shikanov ◽  
...  
Keyword(s):  
Stem Cell ◽  
Liver Tissue ◽  
Drug Testing ◽  
Human Stem Cell

Development of an in vitro stem cell assay for predictive drug testing

1984 ◽  
Vol 17 (3) ◽  
pp. 215
Author(s):  
D.I. Robertson ◽  
G. Stuart ◽  
A. Wong ◽  
P.H. Geggie ◽  
O. Siu
Keyword(s):  
Stem Cell ◽  
Drug Testing ◽  
Cell Assay

scholarly journals A method for human teratogen detection by geometrically confined cell differentiation and migration

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Jiangwa Xing ◽  
Yi-Chin Toh ◽  
Shuoyu Xu ◽  
Hanry Yu
Keyword(s):  
Stem Cell ◽  
Epithelial Mesenchymal Transition ◽  
False Negative ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cell ◽  
Mesenchymal Transition ◽  
Screening Platform ◽  
And Migration ◽  
Species Specific

Abstract Unintended exposure to teratogenic compounds can lead to various birth defects; however current animal-based testing is limited by time, cost and high inter-species variability. Here, we developed a human-relevant in vitro model, which recapitulated two cellular events characteristic of embryogenesis, to identify potentially teratogenic compounds. We spatially directed mesoendoderm differentiation, epithelial-mesenchymal transition and the ensuing cell migration in micropatterned human pluripotent stem cell (hPSC) colonies to collectively form an annular mesoendoderm pattern. Teratogens could disrupt the two cellular processes to alter the morphology of the mesoendoderm pattern. Image processing and statistical algorithms were developed to quantify and classify the compounds’ teratogenic potential. We not only could measure dose-dependent effects but also correctly classify species-specific drug (Thalidomide) and false negative drug (D-penicillamine) in the conventional mouse embryonic stem cell test. This model offers a scalable screening platform to mitigate the risks of teratogen exposures in human.

scholarly journals Neuronal Differentiation of Adipose Derived Stem Cells: Progress So Far

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
T. J. Moore ◽  
Heidi Abrahamse
Keyword(s):  
Stem Cells ◽  
Nervous System ◽  
Stem Cell ◽  
Cell Therapy ◽  
Stem Cell Therapy ◽  
Treatment Modalities ◽  
Adipose Derived Stem Cells ◽  
Positive Effects ◽  
Therapeutic Regime

The nervous system is essential for normal physiological function of all systems within the human body. Unfortunately the nervous system has a limited capacity for self-repair and there are a plethora of disorders, diseases, and types of trauma that affect the central and peripheral nervous systems; however, current treatment modalities are unable to remedy them. Stem cell therapy using easily accessible mesenchymal stem cells, such as those found in the adipose stroma, has come to the fore in a number of biomedical disciplines as a potential therapeutic regime. In addition to substantial research already having been conducted on thein vitrodifferentiation of stem cells for the treatment of neurological repair, numerous strategies for the induction and culture of stem cells into terminal neural lineages have also been developed. However, none of these strategies have yet been able to produce a fully functional descendent suitable for use in stem cell therapy. Due to the positive effects that low level laser irradiation has shown in stem cell studies to date, we propose that it could enhance the processes involved in the differentiation of adipose derived stem cells into neuronal lineages.

scholarly journals Development of a Screening Platform to Identify Small Molecules That Modify ELP1 Pre-mRNA Splicing in Familial Dysautonomia

2018 ◽  
Vol 24 (1) ◽  
pp. 57-67 ◽  
Author(s):  
Monica Salani ◽  
Fabio Urbina ◽  
Anthony Brenner ◽  
Elisabetta Morini ◽  
Ranjit Shetty ◽  
...  
Keyword(s):  
Method Development ◽  
Donor Site ◽  
Screening Method ◽  
Familial Dysautonomia ◽  
Mrna Splicing ◽  
Splice Donor Site ◽  
Signal Ratio ◽  
Screening Platform ◽  
Exon 20

Familial dysautonomia (FD) is an autonomic and sensory neuropathy caused by a mutation in the splice donor site of intron 20 of the ELP1 gene. Variable skipping of exon 20 leads to a tissue-specific reduction in the level of ELP1 protein. We have shown that the plant cytokinin kinetin is able to increase cellular ELP1 protein levels in vivo and in vitro through correction of ELP1 splicing. Studies in FD patients determined that kinetin is not a practical therapy due to low potency and rapid elimination. To identify molecules with improved potency and efficacy, we developed a cell-based luciferase splicing assay by inserting renilla (Rluc) and firefly (Fluc) luciferase reporters into our previously well-characterized ELP1 minigene construct. Evaluation of the Fluc/Rluc signal ratio enables a fast and accurate way to measure exon 20 inclusion. Further, we developed a secondary assay that measures ELP1 splicing in FD patient-derived fibroblasts. Here we demonstrate the quality and reproducibility of our screening method. Development and implementation of this screening platform has allowed us to efficiently screen for new compounds that robustly and specifically enhance ELP1 pre-mRNA splicing.

scholarly journals Comparison of the Simulated Response of Three in Silico Human Stem Cell-Derived Cardiomyocytes Models and in Vitro Data Under 15 Drug Actions

2021 ◽  
Vol 12 ◽  
Author(s):  
Michelangelo Paci ◽  
Jussi T. Koivumäki ◽  
Hua Rong Lu ◽  
David J. Gallacher ◽  
Elisa Passini ◽  
...  
Keyword(s):  
Experimental Data ◽  
Stem Cell ◽  
In Silico ◽  
Drug Testing ◽  
Drug Application ◽  
Human Stem Cell ◽  
Drug Induced ◽  
Drug Actions ◽  
Experimental Findings

Objectives: Improvements in human stem cell-derived cardiomyocyte (hSC-CM) technology have promoted their use for drug testing and disease investigations. Several in silico hSC-CM models have been proposed to augment interpretation of experimental findings through simulations. This work aims to assess the response of three hSC-CM in silico models (Koivumäki2018, Kernik2019, and Paci2020) to simulated drug action, and compare simulation results against in vitro data for 15 drugs.Methods: First, simulations were conducted considering 15 drugs, using a simple pore-block model and experimental data for seven ion channels. Similarities and differences were analyzed in the in silico responses of the three models to drugs, in terms of Ca2+ transient duration (CTD90) and occurrence of arrhythmic events. Then, the sensitivity of each model to different degrees of blockage of Na+ (INa), L-type Ca2+ (ICaL), and rapid delayed rectifying K+ (IKr) currents was quantified. Finally, we compared the drug-induced effects on CTD90 against the corresponding in vitro experiments.Results: The observed CTD90 changes were overall consistent among the in silico models, all three showing changes of smaller magnitudes compared to the ones measured in vitro. For example, sparfloxacin 10 µM induced +42% CTD90 prolongation in vitro, and +17% (Koivumäki2018), +6% (Kernik2019), and +9% (Paci2020) in silico. Different arrhythmic events were observed following drug application, mainly for drugs affecting IKr. Paci2020 and Kernik2019 showed only repolarization failure, while Koivumäki2018 also displayed early and delayed afterdepolarizations. The spontaneous activity was suppressed by Na+ blockers and by drugs with similar effects on ICaL and IKr in Koivumäki2018 and Paci2020, while only by strong ICaL blockers, e.g. nisoldipine, in Kernik2019. These results were confirmed by the sensitivity analysis.Conclusion: To conclude, The CTD90 changes observed in silico are qualitatively consistent with our in vitro data, although our simulations show differences in drug responses across the hSC-CM models, which could stem from variability in the experimental data used in their construction.

Urine Cells-derived iPSCs: An Upcoming Frontier in Regenerative Medicine

2021 ◽  
Vol 28 ◽  
Author(s):  
Sanjeev Gautam ◽  
Sangita Biswas ◽  
Birbal Singh ◽  
Ying Guo ◽  
Peng Deng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Drug Testing ◽  
Diagnostic Tools ◽  
Cell Therapies ◽  
Non Invasive ◽  
Key Issues ◽  
Induced Pluripotent

: There is a momentous surge in the development of stem cell technology as a therapeutic and diagnostic tools. Stem cell-derived cells are currently used in various clinical trials. However, key issues and challenges involve the low differentiation efficiency, integration, and functioning of transplanted stem cells-derived cells. Extraction of bone marrow, adipose, or other mesenchymal stem cells (MSCs) involves invasive methods, specialized skills, and expensive technologies. Urine-derived cells, on the other hand, are obtained by non-invasive methods. Samples can be obtained repeatedly from patients of any age. Urine-derived cells are used to generate reprogrammed or induced pluripotent stem cells (iPSCs), which can be cultured, and differentiated into various types of cell lineages for biomedical investigations and drug testing in vitro or in vivo using model animals of human diseases. Urine cell-derived iPSCs (UiPSCs) have emerged as a major area of research and immense therapeutic significance. Given that preliminary preclinical studies are successful in terms of safety and as a regenerative tool, the UiPSCs will pave the way to develop and expedite various types of autologous stem cell therapies.

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