Therapeutic Potential of Designer Receptors, Gary Aston-Jones, PhD

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  • 1. Different types of neurons are interdigitated in the same brain areas: How selectively stimulate/inhibit? Guan et al, Intl J Obesity, 2002.
  • 2. Gene transfer in brain in vivo viral vectors Davidson & Breakefield, Nat. Rev. Neurosci. 2003.
  • 3. Cell-type-specific gene transfer & modulation of neuronal activity Calloway, E., Trends Neurosci. 2005. Allostatin (AL) = invertebrate peptide/receptor system
  • 4. Evolving the lock to fit the key: Designer receptors exclusively activated by designer drugs (DREADDS) Conklin, B., PNAS (2007) Armbruster et al., PNAS (2007) directed evolution: muscarinic ACh receptor -> DREADD CNO=clozapine n-oxide Muscarinic Ach receptor DEADD
  • 5. Courtesy of Bryan Roth, U North Carolina Specific neuronal inhibition with hM4D DREADD
  • 6. Lentivirus to drive DREADD via orexin promoter: selective activation or inhibition of orexin neurons in behaving animals
    • Brain orexin/hypocxretin system
      • Arousal, stress
      • Reward, addiction
      • Eating, obesity
  • 7. Photosensitive ion channels for control of neuronal activity Zhang et al., Nature Rev. Neurosci. (2007) Channelrhodopsin Halorhodopsin excitation inhibition
  • 8. Photostimulation yields more specific effects than electrical stimulation Alternative to Deep Brain Stimulation Zhang et al., Nature Rev. Neurosci. (2007)
  • 9. Cell-type specific expression of channelrhodopsin in brain norepinephrine neurons in vivo PRSx8 promoter; mCherry tag Microinjection in vivo in locus coeruleus TH (NE neurons) mCherry (ChR2 neurons) Merge David Moorman Elena Vazey
  • 10. Photostimulation of CNS neurons in vivo record laser
  • 11. Photostimulation of neuronal activity in vivo Locus coeruleus neuronal recordings in anesthetized rat David Moorman Elena Vazey
  • 12. Summary
    • Novel proteins designed and expressed in neurons
      • Designer receptors
      • Photosensitive channels/ion pumps
    • Neurons transduced in vivo to express novel proteins
      • Viral vectors (lenti virus, adeno-associated virus, etc)
      • Cell-type specific expression using promoters
    • Many clinical applications
      • Designer receptors (DREADDS): More specific than conventional pharmacology
      • Photostimulation: More specific, less toxic than Deep Brain Stimulation (DBS)
      • Much better targeting ability than conventional approaches, e.g., DBS, lesion
      • Many fewer side-effects
  • 13. Acknowledgements Rachel Smith David Moorman Angie Cason Pouya Tashili-Fahadan Kimberlei Richardson Steve Mahler Lana Zhang Greg Sartor Elena Vazey Mike Smith Becky Fallon Glenda Harris (U Penn) Matt Wimmer (U Penn) MUSC Neuroscience of Addiction Research Center National Institute on Drug Abuse Lilly Pharmaceuticals
  • 14. Transfer Vector Design
    • Promoters
      • mPGK-1
      • hSyn-1: neurons
      • hGFAP: glia
      • CaM kinase II
      • rMOR
      • rPPE
    • Internal ribosome entry sites (IRES)
      • Encephalomyocarditis virus (ECMV)
      • Synthetic (Gtx)
    • Woodchuck hepatitis virus post-translational regulatory element (WPRE)
    Promoter Transgene IRES EGFP WPRE