Poster CC12 - Neuron-specific cytokine signature of MSC-induced neuroprotection in a cell culture model of Parkinson’s Disease

Friday, October 25, 2024

3:30 PM - 4:30 PM EST

Location: Exhibit Hall E, F & G

Presenting Author(s)

AH

Abigail L. Holberton (she/her/hers)

Research Technician
George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology
Marietta, Georgia, United States

Co-Author(s)

RC

Raquel Caldeira Botelho

Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, United States
Ariana Kalantari

Undergraduate Researcher
School of Biological Sciences, Georgia Institute of Technology
Atlanta, Georgia, United States
KZ

Kevin Zheng

Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, United States
Adhrija Anbuchelian (she/her/hers)

Research Assistant
School of Biological Sciences, Georgia Institute of Technology
Cumming , Georgia, United States
SD

Suzann Donovan

Department of Neuroscience, Emory University, United States

Primary Investigator(s)

Levi B. Wood, PhD

Associate Professor
George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology
Atlanta, Georgia, United States

Last Author(s)

Melissa L. Kemp, PhD

Professor
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology
Atlanta, Georgia, United States

Introduction: Parkinson's disease (PD) is a neurodegenerative disease characterized by the loss of dopaminergic (DA) neurons and accumulation of alpha synuclein aggregates, but mechanisms of neurodegeneration and methods to promote neuroprotection remain poorly understood. Mitochondrial dysfunction and neuronal death are central cellular phenotypes in PD. Mesenchymal stem cells (MSCs) have recently been shown to provide cellular protection in pathogenic environments as well as transfer their own healthy mitochondria into neurons. Here, we asked if MSCs would transfer mitochondria and modify neuronal function in the presence of aggregated alpha synuclein pre-formed fibrils (PFFs). To test this, and define the effects of MSC on neurons, we utilized the N2A neuronal cell line transduced with the biotin ligase TurboID tag to assess the N2A-specific effects of the presence of MSCs. We used cytokine expression as our primary readout because of the importance of mitochondria in regulating the immune microenvironment. We hypothesized that MSCs would modulate N2A cytokine secretion into the culture medium in the presence of PFFs.

Materials and

Methods: TurboID N2As were conditioned with PFFs and biotin for 24 hours and then co-cultured with MSCs for an additional 24 hours. An adapted Luminex multiplexed ELISA was used to quantify biotinylated proteins secreted by the N2As from the conditioned media to identify key contributors in the neuronal response to PFFs and co-culture environment.

Results, Conclusions, and Discussions: When conditioned with alpha synuclein PFFs, the TurboID N2As with MSCs had an increase in neuroprotective and anti-inflammatory cytokine expression, including IL-15 and IL-10 (Figure 1, Green box, Dunn’s test, p< 0.05), when compared to an N2A mono-culture (Figure 1, Yellow box), highlighting potentially protective stimulating effects of the MSCs on N2A function. Moreover, these results demonstrate the ability of using TurboID to deconvolute cell type specific signaling in a co-culture systems. Future studies in transgenic TurboID mice will illuminate the DA neuron response to PFFs in whole tissue and enable evaluation of MSCs as a stimulator of neuroprotective and anti-inflammatory signaling.

Acknowledgements (Optional):