# Characterization of CNS protein aggregate components during acute and chronic EAE

> **NIH NIH R21** · ALBERT EINSTEIN COLLEGE OF MEDICINE · 2020 · $461,389

## Abstract

Multiple sclerosis (MS) is a multifocal demyelinating disease that results in neural dysfunction and culminates
in physical and cognitive disability. During MS, immune cells attacks the myelin sheath causing inflammation,
axonal damage, and impaired remyelination. Repair is inefficient in the presence of misfolded proteins, protein
aggregates, damaged organelles, and myelin debris, resulting in chronic disability and neurodegeneration.
Therefore, it is important to define the damaged cellular components and understand how to efficiently
eliminate them from CNS lesions that develop during MS. Protein aggregates are highly resistant to protein
degradation and targeting them for clearance is challenging. The long-term goal is to identify the proteins
within aggregates and understand how to eliminate them for efficient clearance of debris and better repair.
Cells attempt to remove misfolded proteins by degrading, refolding, or sequestering them in intracellular
compartments. If the proteins and the mechanisms by which they are degraded are identified, the efficiency of
debris clearance can be improved with targeted therapy. In this proposal, we will test the hypothesis that
following sensitization with MOG35-55 neuronal/axonal proteins and proteins enriched in the myelin
sheath are inefficiently cleared and form aggregates. The rationale for this project is the observation that
protein of unknown composition form aggregates in MS lesions. The central hypothesis will be tested by
pursuing two specific aims. Specific Aim 1 will characterize and validate the proteins within aggregates during
acute and chronic EAE. As part of our study, we will compare protein aggregates from age- and sex-matched
mouse spinal cord and identify the proteins by mass spectrometry (nanoLC-mass spectrometry ms/ms). This
analysis will identify common functions and conserved motifs within the identified proteins and compare
functions and clearance pathways using bioinformatics analyses, and western blot analysis. Specific Aim 2
will examine how clearance by the proteasome and macroautophagy (autophagy) affects protein aggregate
accumulation. Treatment with a disaccharide shown to activate autophagy will determine whether treatment is
able to reduce the number of proteins associated with aggregates as well as reduce clinical scores of treated
mice. The research proposed is innovative because it provides a framework to identify and assess proteins
and the signaling pathways associated with insoluble aggregates observed in EAE. The proposed research is
significant because it is expected to provide scientific justification to address how to clear protein aggregates,
improve repair, and identify target areas for therapy designed to efficiently clear cellular clearance and reduce
the duration in which axons are exposed to an unfavorable milieu in the CNS.

## Key facts

- **NIH application ID:** 9955591
- **Project number:** 1R21NS116526-01
- **Recipient organization:** ALBERT EINSTEIN COLLEGE OF MEDICINE
- **Principal Investigator:** BRIDGET SHAFIT-ZAGARDO
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $461,389
- **Award type:** 1
- **Project period:** 2020-05-01 → 2022-10-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9955591

## Citation

> US National Institutes of Health, RePORTER application 9955591, Characterization of CNS protein aggregate components during acute and chronic EAE (1R21NS116526-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9955591. Licensed CC0.

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