Project description:Here, we show that midbrain dopamine (DA) neurons derived from human pluripotent stem cells (hPSCs) are selectively permissive to SARS-CoV-2 infection both in vitro and upon transplantation in vivo, and that SARS-CoV-2 infection triggers a DA neuron inflammatory and cellular senescence response. A high-throughput screen in hPSC-derived DA neurons identified several FDA approved drugs, including riluzole, metformin, and imatinib, that can rescue the cellular senescence phenotype and prevent SARS-CoV-2 infection. RNA-seq analysis of human ventral midbrain tissue from COVID-19 patients, using formalin-fixed paraffin-embedded autopsy samples, confirmed the induction of an inflammatory and cellular senescence signature and identified low levels of SARS-CoV-2 transcripts. Our findings demonstrate that hPSC-derived DA neurons can serve as a disease model to study neuronal susceptibility to SARS-CoV-2 and to identify candidate neuroprotective drugs for COVID-19 patients. The susceptibility of hPSC-derived DA neurons to SARS-CoV-2 and the observed inflammatory and senescence transcriptional responses suggest the need for careful, long-term monitoring of neurological problems in COVID-19 patients.
Project description:COVID-19 patients commonly present with neurological signs of central nervous system (CNS) and/or peripheral nervous system dysfunction. However, which neural cells are permissive to infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been controversial. Here, we show that midbrain dopamine (DA) neurons derived from human pluripotent stem cells (hPSCs) are selectively susceptible and permissive to SARS-CoV-2 infection, and that SARS-CoV-2 infection triggers a DA neuron inflammatory and cellular senescence response. A high-throughput screen in hPSC-derived DA neurons identified several FDA approved drugs, including riluzole, metformin, and imatinib, that can rescue the cellular senescence phenotype by preventing SARS-CoV-2 infection. RNA-seq analysis of human ventral midbrain tissue from COVID-19 patients, using formalin-fixed paraffin-embedded autopsy samples, confirmed the induction of an inflammatory and cellular senescence signature and identified low levels of SARS-CoV-2 transcripts. Our findings demonstrate that hPSC-derived DA neurons can serve as a disease model to study neuronal susceptibility to SARS-CoV-2 and to identify candidate neuroprotective drugs for COVID-19 patients. The susceptibility of hPSC-derived DA neurons to SARS-CoV-2 and the observed inflammatory and senescence transcriptional responses suggest the need for careful, long-term monitoring of neurological problems in COVID-19 patients.
Project description:COVID-19 patients commonly present with signs of central nervous system and/or peripheral nervous system dysfunction. Here, we show that midbrain dopamine (DA) neurons derived from human pluripotent stem cells (hPSCs) are selectively susceptible and permissive to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. SARS-CoV-2 infection of DA neurons triggers an inflammatory and cellular senescence response. High-throughput screening in hPSC-derived DA neurons identified several FDA-approved drugs that can rescue the cellular senescence phenotype by preventing SARS-CoV-2 infection. We also identified the inflammatory and cellular senescence signature and low levels of SARS-CoV-2 transcripts in human substantia nigra tissue of COVID-19 patients. Furthermore, we observed reduced numbers of neuromelanin+ and tyrosine-hydroxylase (TH)+ DA neurons and fibers in a cohort of severe COVID-19 patients. Our findings demonstrate that hPSC-derived DA neurons are susceptible to SARS-CoV-2, identify candidate neuroprotective drugs for COVID-19 patients, and suggest the need for careful, long-term monitoring of neurological problems in COVID-19 patients.
Project description:RNA-seq analysis was applied to examine the mock or SARS-CoV-2 infected DA neurons.We found SARS-CoV-2 infection caused DA neurons senescence.We also found several drug candidates block SARS-CoV-2 infection caused senescence of hPSC-derived DA neurons. RNA-seq was applied to analyze the drug candidates or DMSO treated DA neurons upon SARS-CoV-2 infection. The genes involved in senescence pathway were rescued in riluzole, metformin or imatinib treated DA neurons.
Project description:Transcriptomic analysis of enriched of neuron population collected from in-vivo mouse brains has been a challenge in the neuroscience field due to its fragility in withstanding harsh condition during isolation and collection process. We established a fluorescent reporter mouse Ddc-hKO1 to facilitate the identification and collection of Ddc-expressing neurons (dopaminergic, serotonergic, cholinergic and adrenergic neurons) by the detection of red fluorescence signals using a FACs. We utilized an improved isolation protocol that ensure high yield and quality of viable neurons during collection that is suitable for RNA-sequencing. This is the first report of transcriptomic profiling of Ddc expressing (hKO1(+)) neurons. Successful collection of Ddc-expressing neurons were verified by gene markers of dopaminergic, serotonergic, cholinergic neurons in hKO1(+) populations while other neuron types in hKO1(-) population. Furthermore, GSEA analysis were performed on both hKO1(+) and hKO1(-) to further support the neuron types collected in respective population.
Project description:A mounting body of evidence reveals that inflammation is a major hallmark in the pathophysiology of Parkinson’s disease (PD). Notably, microglia are key players in orchestrating neuroinflammatory signalling pathways in the brain. However, the molecular mechanisms underlying the microglial response in the dopaminergic neuron degeneration remain unclear. In this study, we investigated the role of the PD-associated LRRK2-G2019S variant in microglial neurotoxicity that potentially drives the loss of dopaminergic neurons. Using PD patient-specific induced pluripotent stem cell-based models, we showed that microglia harbouring the LRRK2-G2019S mutation exhibited increased activation markers, phagocytic capacity, and secretion of inflammatory cytokines such as TNF-α. This was linked to metabolic dysregulation, including higher levels of glycolysis and a reduction in serine biosynthesis. These overactivated microglia drove dopaminergic neuron degeneration in a 3D midbrain organoid model. Intriguingly, addressing the PD associated metabolic disturbances reduced the inflammatory state of the microglia and reversed neuronal loss. These findings reinforce the immunometabolic control of microglia to preserve dopaminergic neuron integrity and its pivotal role in PD pathogenesis.
Project description:Performed RNA-seq analysis of animals with xbp-1s overexpression (ER stress response transcription factor) in specific neuron types: pan-neuronal, serotonergic neuron, dopaminergic neuron, and both serotonergic and dopaminergic neurons, all compared to a wild-type control. RNA-seq was performed on purified RNA extracted from ~1000 whole worms using a proprietary Genewiz protocol described briefly in the manuscript. 3 biological replicates are provided for each sample.