Characterization of neural crest stem cells for a DPN-specific treatment

Grant Winners

Deans

Abstract

Diabetic peripheral neuropathy (DPN), a term that encompasses the damage of peripheral nerves-in particular those found in the extremities-is the most frequently observed serious and chronic complication of diabetes. With a prevalence of 30-50% among patients with diabetes, >70% of those with DPN describe it as severely painful, significantly impacting their quality of life. While some studies using pharmacological treatments have proven successful, pharmacological treatments are described as "disappointing," and provide no improvement for >35% of patients with DPN. The quality of life destroying effects of DPN and severe lack of success using pharmaceuticals underscore the dire need for a specific, long-term DPN treatment. In DPN, there are changes in the peripheral and central nervous systems. An increase in the release of the excitatory neurotransmitter glutamate and in the amount of its receptor may lead to excessive nerve firing in DPN. Additionally, there is the death of GABAergic interneurons (responsible for inhibiting nerve firing) and the decrease in their receptors. Together, this is known as central sensitization. The objective of this application is to characterize neural crest derived stem cells (NCSCs) from dental tissues to determine their therapeutic potential for DPN. The central hypothesis is that NCSCs will be amenable to neuro-differentiation and display more desirable properties than other stem cells. Our hypothesis is formulated on the basis that stem cells have been differentiated towards a GABAergic lineage. The rationale for the proposed research is that a DPN treatment may be achieved through characterization and differentiation, which will create a micro-environment that will arrest/reduce the effects of DPN in-vivo. We plan to test our central hypothesis and attain the objective of this application by pursuing the following specific aims: 1) Establish and characterize NCSC lines from various dental tissues and 2) Develop a neuro-differentiation protocol for the NCSC lines, including optimization of length of treatment and growth factor combination. In Aim 1, we will purify stem cells and use immunohistochemical (IHC) and genetic analysis to verify the resulting NCSCs. In Aim 2, we will neuro-differentiate NCSCs and use IHC, genetic, and neural assays to examine the effects of differentiation. Our research is innovative and significant because it is geared towards a DPN-specific treatment, is based on an etiology of DPN and central sensitization, and uses an approach geared towards reducing time from bench to bedside. This research may lead to a novel stem cell-based treatment for DPN.