The development of new therapies to treat amyotrophic lateral sclerosis (ALS) is often hindered by the poor bioavailability of candidate drugs to affected motor neurons within the central nervous system (CNS). Thus, while both glial cell line-derived neurotrophic factor (GDNF) and insulin-like growth factor-1 (IGF-1) have had robust survival promoting effects on injured motor neurons in experimental animals, their success in treating ALS patients appears to have been thwarted by insufficient access to motor neurons in the much larger human CNS. We hypothesize that genetic or chemical fusion of tetanus toxin fragment C (TTC) to either GDNF or IGF-1 will improve growth factor delivery to motor neurons through one or more mechanisms related to the nerve cell binding properties of TTC. The primary goal of this project, then, is to assess whether the neuroprotective activity of IGF-1:TTC and GDNF:TTC in an experimental animal model of ALS is superior to that of the respective growth factor alone. Our study has four specific aims: (1) optimize the design, expression, and purification of IGF-1:TTC and GDNF:TTC; (2) characterize the purity, stability, and basic functional activity of these fusion proteins; (3) examine the bioavailability and functional activity of the fusion proteins in rodent CNS following intracerebroventricular or intramuscular administration; and (4) investigate the neuroprotective activity of the fusion proteins in a transgenic rat model of amyotrophic lateral sclerosis (ALS). The experiments in Aim 1 will use recombinant protein expression and purification techniques to generate the fusion proteins, while the studies in Aims 2 and 3 will employ immunocytochemical, Western blot, enzyme immunoassay, and morphometric techniques to assess the functional properties of GDNF:TTC and IGF-1:TTC in cultured ceils and whole animals. Finally, the experiments in Aim 4 will employ transgenic animals in survival and behavioral experiments to assess the neuroprotective effects of GDNF:TTC and/or IGF-1 :TTC in vivo. While the present work may give rise to a new treatment for ALS, the information obtained from our characterization of IGF-1:TTC and GDNF:TTC may lead to the use of these fusion proteins in other neurological disorders as well.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS038679-06
Application #
7009596
Study Section
Clinical Neuroplasticity and Neurotransmitters Study Section (CNNT)
Program Officer
Tagle, Danilo A
Project Start
1999-04-19
Project End
2008-12-31
Budget Start
2006-01-01
Budget End
2006-12-31
Support Year
6
Fiscal Year
2006
Total Cost
$647,939
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
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