Investigating the relationship between mitochondrial deficits and proteostasis dysfunction in cellular models of Parkinson’s disease
For the past two years I have been a research assistant in the Lavoie Lab of the Center for Translational Research in Neurodegenerative Diseases within the College of Medicine at the University of Florida. In June 2024 I began work on a new project under the supervision of my mentor Dr. Nitya Subrahmanian, Research Asst. Professor, and our lab PI and CTRND Director Dr. Matthew J. LaVoie. The aim of my project is to investigate whether mild deficits in Complex I of the electron transport chain are sufficient to promote pathological α-synuclein accumulation in neuronal models of Parkinson's Disease.
Biological Safety Cabinet where I perform cell culture work
Example of an immunoblot I ran for NDUFAF2 expression in Human Embryonic Kidney cells
Research Focus
The focus of this project is to determine the effect that knocking down the expression of NDUFAF2, an assembly factor of Complex I, has on the accumulation of alpha synuclein. Aggregation this protein is assumed to be the main contributor to the death of dopaminergic cells, and the progressive loss of these cells is what causes the motor and nonmotor collection of symptoms that make up Parkinson's Disease. Preliminary data from the LaVoie Lab points to mild Complex I deficits of the ETC as being a potential driver of synuclein aggregation in idiopathic PD, and I seek to replicate this phenomenon by knocking down NDUFAF2 expression in stem cell derived neurons, thereby partially inhibiting CI activity, and investigating the downstream impact this has on synuclein aggregation.
Project Aims and Responsibilities
I have several aims for this project. my preliminary goal is to use lentiviral transduction to knockdown NDUFAF2 expression with shRNA in induced Pluripotent Stem Cells. Once I confirm successful knockdown via immunoblotting, I will then generate induced neurons from these iPSCs. With a neuronal model established, I will then focus on answering two questions. Aim one to determine the effect mild CI deficits have on the accumulation of exogenously introduced synuclein. I will assess this using the complementary methods of immunocytochemistry and immunoblotting. Aim two is to identify what pathological species of truncated synuclein these iNs generate in response to endogenous synuclein stress. Through these aims I hope to answer important questions about mitochondrial stress in relation to Parkinson's Disease.