Intranasal Delivery of Proteins Using Cationic Liposomes for the Treatment of Parkinson’s Disease and the Use of Bioquant ® Image Analysis Software Presented by Mattia M. Migliore April 20, 2007

Introduction:  Parkinson’s 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.

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.

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.

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 Parkinson’s disease.

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.

 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.

Using Bioquant ® to Map Protein Distribution in the Brain Olfactory Bulb

Using Bioquant ® to determine co-localization of the administered protein with a dopamine neuronal marker, tyrosine hydroxylase Conditional Frequency Analysis

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.

Conclusion:  Bioquant ® will be used to qualitatively and quantitatively analyze the data in this project.