Year of Publication

2013

Degree Name

Doctor of Philosophy (PhD)

Document Type

Doctoral Dissertation

College

Medicine

Department

Anatomy and Neurobiology

First Advisor

Dr. Luke Bradley

Second Advisor

Dr. Don Gash

Abstract

Parkinson’s disease, the second most common neurodegenerative disease, is characterized by the loss of dopaminergic neurons within the substantia nigra. Currently, the treatments available for PD are symptomatic treatments that do not stop the progression of the disease. Trophic molecules, such as glial cell-line derived neurotrophic factor (GDNF), have been evaluated as potential therapeutic molecules that could stop the loss of neurons and potentially restore some of the neurons that have already been lost. However, these trophic molecules are large making them difficult to produce and delivery. Here we characterize three peptides (DNSP-5, DNSP-11, and DNSP-17) to determine it they are stable and offer protective effects similar to GNDF allowing them to be potential therapeutic molecules.

The data presented here involves the evaluation of the molecular and cellular mechanism of DNSP-5, DNSP-11, and DNSP-17, which are derived from prosequence of GDNF. Initial studies were carried out to evaluate the physical characteristics of these three peptides to determine their viability as potential therapeutic molecules. The structure and stability of these peptides were evaluated. Based on the data it was determined that the three peptides do not interact in vitro, allowing for further individual evaluations of the peptides. It was also determined that the peptides were stable when stored at both -80°C and 37°C for one month, allowing them to both potentially be stored during treatment.

Cell culture assays and proteomic profiling were utilized to determine binding partners and potential mechanisms through which DNSP-11 may be able to mediate apoptosis. It was determined that DNSP-11 was able to interact with a variety of binding partners that are involved in metabolism. These studies have aided in the understanding of neurotrophic factor prosequence function, but will also serve as a starting point for the development of novel trophic factors for PD treatment.

Finally, the interaction between DNSP-11 and GAPDH was evaluated as a potential anti-apoptotic mechanism. GAPDH has previously shown to play a role in mediating apoptotic pathways. It was hypothesized that the observed interaction between DNSP-11 and GAPDH could mediate that role of GAPDH in apoptosis and afford DNSP-11 its observed anti-apoptotic effects. It was observed that while DNSP-11’s interaction with GAPDH may play a role in its anti-apoptotic effects, it does not appear to be the only mechanism involved. Based on this data, it is likely that the other metabolic binding partners play a role in DNSP-11’s anti-apoptotic mechanisms and therefore, these interactions should be further evaluated.

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