Intranasal Delivery of Proteins Using Cationic Liposomes for the Treatment of Parkinson

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Intranasal Delivery of Proteins Using Cationic
Liposomes for the Treatment of Parkinsons
Disease and the Use of Bioquant Image Analysis
Software

• Presented by Mattia M. Migliore
• April 20, 2007

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Introduction

• Parkinsons disease (PD) is a progressive
  neurodegenerative disease, which interferes with
  normal motor function, and eventually results in
  akinesia and death.
• Results from the destruction of dopaminergic
  neurons of the A9 nigrostriatal pathway.
• Affects approximately 1.5 million people in the
  US alone.
• PD has no cure and current treatments only
  provide temporary symptomatic relief.

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Introduction (cont.)

• GDNF is a protein with therapeutic potential for
  PD because it exerts neurotrophic and
  neuroregenerative effects of dopamine neurons.
• GDNF levels are decreased by as much as 19.4 per
  SN neuron in PD patients (Chauhan et al., 2001
  Hurelbrink and Barker, 2004).
• GDNF does not cross the blood-brain barrier
  (BBB).
• GDNF administration requires invasive
  intracerebral infusions to reach its site of
  action.

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Introduction (cont.)

• The goal of this project is to develop a cationic
  liposomal drug delivery system to transport GDNF
  to the brain using the intranasal route of
  administration.
• The intranasal route of administration was chosen
  because it is non-invasive, and it bypasses the
  BBB.

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Specific AIMS

• Specific AIM 1 To characterize and optimize a
  nanoparticle formulation for intranasal GDNF.
• Using first a model protein to optimize our
  cationic liposomal formulation.
• Specific AIM 2 To determine brain delivery of
  GDNF in rats following intranasal administration.
• Specific AIM 3 To determine the therapeutic
  efficacy of intranasal GDNF in a rat model of
  Parkinsons disease.

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Using Bioquant to Quantitate Protein Brain
Delivery

• Fluorescently tagged ovalbumin was intranasally
  administered to rats.
• The fluorescent label, Alexa-488 was seen
  intracellularly in coronal brain sections.

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• Quantification was accomplished by thresholding
  the cells that take up the protein and performing
  a pixel count.
• GDNF immunohistochemistry was performed w/ a
  fluorescently tagged TxR secondary antibody
  following the same procedure.

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Using Bioquant to Map Protein Distribution in
the Brain
Olfactory Bulb
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Using Bioquant to determine co-localization of
the administered protein with a dopamine
neuronal marker, tyrosine hydroxylase
Conditional Frequency Analysis
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Using Bioquant to Determine If GDNF Can
Effectively Protect Against a 6-Hydroxydopamine
Lesion

• 6-Hydroxydopamine will be injected into the MFB
  to create an animal model of PD.
• Bioquant will be used to quantify the extent of
  the lesion, with a goal unilateral lesion of
  50-75.
• Following administration of GDNF, we will
  quantify the lesion to look for therapeutic
  effectiveness.

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Conclusion

• Bioquant will be used to qualitatively and
  quantitatively analyze the data in this project.