Development of a Biosensor-Toolbox to Perform Chemical and Genetic Perturbance Screens in Mouse Neurons and Cellular Models of Psychiatric Diseases

Dienstagskolloquium

  • Datum: 31.05.2016
  • Uhrzeit: 15:00 - 16:00
  • Vortragende(r): Prof. Dr. Moritz Rossner
  • Neurobiologie psychiatrischer Erkrankungen, Klinik für Psychiatrie und Psychotherapie der LMU München, Campus Innenstadt
  • Ort: Max-Planck-Institut für Psychiatrie
  • Raum: Hörsaal
  • Gastgeber: Jan Deussing
Development of a Biosensor-Toolbox to Perform Chemical and Genetic Perturbance Screens in Mouse Neurons and Cellular Models of Psychiatric Diseases

Psychiatric diseases such as schizophrenia, bipolar disorder and autism spectrum disorders
are considered neurodevelopmental synaptopathies. Compelling evidence has been obtained
from large-scale genome-wide association studies which identified a plethora of genetic
variations within hundreds of genes encoding components of the post-synaptic compartment
and cellular as well as nuclear calcium signaling mediating excitation-transcription coupling.
There has been a tremendous progress in neuroscience research within the last decade to
employ an array of molecular and cellular techniques studying the impact of individual genes.
Neuroscience is, however, methodologically lagging behind the cancer field with respect to
large scale functional chemical and genetic screens. The difficulties of culturing postmitotic
and matured neurons in sufficiently large amounts and the development of robust readouts for
high-throughput screening are likely among the most important underlying reasons.
To adress some of these limitations, we have developed a functional genomics toolbox that is
applicable in primary neurons to screen for thousands of chemical or genetic modulators. The
sensitivity and robustness of the screen is largely driven by the pre-selection of genetic
reporters that respond to neuronal activity with high signal-to-noise ratios.
A proof-of-principle will be presented in which an AAV-based RNAi library was screened for
regulators of neuronal excitation and synapse-to-nucleus signaling using a genetic sensor
based on the Synaptic-Activity-Response-Element (SARE). The assay principle relies on
molecular barcodes which serve as quantitative reporters and at the same time as unique
identifiers of the targeted genes. Upon synaptic stimulation, the screen identified hundreds of
genes known to be involved in glutamatergic synapse-to-nucleus signaling including many
regulators of calcium- and cAMP-signaling. Reproducibility of the technical approach has been
verified by a substantial overlap of hits between three independent screens. Moreover, the
SARE reporter also allowed us to perform a screen with nearly 1k of clinically approved
compounds in which several potential negative and positive modulators of neuronal and/or
synaptic activity including several neuroleptics and anti-depressants were identified.
The presented genetic toolbox can be applied for the analysis of regulatory processes in
mouse primary neurons and is currently evaluated for usage in human cellular disease models

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