Transcript Intranasal Delivery of GDNF for the Treatment of Parkinson

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.