ABSTRACT
Tauopathies are age-associated neurodegenerative diseases whose mechanistic underpinnings remain elusive, partially due to a lack of appropriate human models. Here, we engineered human induced pluripotent stem cell (hiPSC)-derived neuronal lines to express 4R Tau and 4R Tau carrying the P301S MAPT mutation when differentiated into neurons. 4R-P301S neurons display progressive Tau inclusions upon seeding with Tau fibrils and recapitulate features of tauopathy phenotypes including shared transcriptomic signatures, autophagic body accumulation, and reduced neuronal activity. A CRISPRi screen of genes associated with Tau pathobiology identified over 500 genetic modifiers of seeding-induced Tau propagation, including retromer VPS29 and genes in the UFMylation cascade. In progressive supranuclear palsy (PSP) and Alzheimer's Disease (AD) brains, the UFMylation cascade is altered in neurofibrillary-tangle-bearing neurons. Inhibiting the UFMylation cascade in vitro and in vivo suppressed seeding-induced Tau propagation. This model provides a robust platform to identify novel therapeutic strategies for 4R tauopathy.
Subject(s)
Induced Pluripotent Stem Cells , Neurons , Tauopathies , tau Proteins , Humans , Induced Pluripotent Stem Cells/metabolism , tau Proteins/metabolism , Tauopathies/metabolism , Tauopathies/pathology , Neurons/metabolism , Neurons/pathology , Animals , Mice , Alzheimer Disease/metabolism , Alzheimer Disease/pathology , Alzheimer Disease/genetics , Brain/metabolism , Brain/pathology , Supranuclear Palsy, Progressive/metabolism , Supranuclear Palsy, Progressive/pathology , Supranuclear Palsy, Progressive/genetics , Cell Differentiation , Mutation , AutophagyABSTRACT
Alphaherpesvirinae family members can reactivate from latency following stress. The synthetic corticosteroid dexamethasone induces certain cellular transcription factors in murine and bovine trigeminal ganglionic neurons. Three dexamethasone-induced transcription factors, Krüppel-like factor 15, Slug, and SPDEF, stimulated the herpes simplex virus type 1-infected cell protein 0 (ICP0) promoter more than 150-fold. Conversely, other viral promoters (VP16 and ICP4) were not strongly stimulated, suggesting that the ICP0 promoter is preferentially activated by dexamethasone-simulated stress.
Subject(s)
Herpes Simplex/metabolism , Herpesvirus 1, Human/genetics , Immediate-Early Proteins/genetics , Kruppel-Like Transcription Factors/metabolism , Nuclear Proteins/metabolism , Promoter Regions, Genetic , Proto-Oncogene Proteins c-ets/metabolism , Transcription Factors/metabolism , Trigeminal Ganglion/metabolism , Ubiquitin-Protein Ligases/genetics , Animals , Dexamethasone/pharmacology , Female , Gene Expression Regulation, Viral , Herpes Simplex/genetics , Herpes Simplex/physiopathology , Herpes Simplex/virology , Herpesvirus 1, Human/metabolism , Humans , Immediate-Early Proteins/metabolism , Kruppel-Like Transcription Factors/genetics , Mice , Neurons/drug effects , Neurons/metabolism , Neurons/virology , Nuclear Proteins/genetics , Proto-Oncogene Proteins c-ets/genetics , Snail Family Transcription Factors , Stress, Physiological , Transcription Factors/genetics , Trigeminal Ganglion/drug effects , Trigeminal Ganglion/virology , Ubiquitin-Protein Ligases/metabolism , Up-RegulationABSTRACT
Nora virus is a recently discovered RNA picorna-like virus that produces a persistent infection in Drosophila melanogaster, but the antiviral pathway or change in gene expression is unknown. We performed cDNA microarray analysis comparing the gene expression profiles of Nora virus infected and uninfected wild-type D. melanogaster. This analysis yielded 58 genes exhibiting a 1.5-fold change or greater and p-value less than 0.01. Of these genes, 46 were up-regulated and 12 down-regulated in response to infection. To validate the microarray results, qRT-PCR was performed with probes for Chorion protein 16 and Troponin C isoform 4, which show good correspondence with cDNA microarray results. Differential regulation of genes associated with Toll and immune-deficient pathways, cytoskeletal development, Janus Kinase-Signal Transducer and Activator of Transcription interactions, and a potential gut-specific innate immune response were found. This genome-wide expression profile of Nora virus infection of D. melanogaster can pinpoint genes of interest for further investigation of antiviral pathways employed, genetic mechanisms, sites of replication, viral persistence, and developmental effects.
