Phosphoproteomic profiling of bryostatin-1-modulated CNS innate immunity for MS therapeutic target discovery

Overview

This project investigates how bryostatin-1, a brain-penetrant compound, modulates protein kinase C (PKC) activity in central nervous system (CNS) innate immune cells, including microglia and CNS-associated macrophages. Prior studies have shown that bryostatin-1 stimulates an anti-inflammatory and regenerative phenotype in these cells, promoting immunomodulation and remyelination. However, the specific proteins phosphorylated downstream of PKC activation remain unknown. By profiling the phosphoproteome, this research aims to identify critical signaling molecules involved in neuroinflammation and remyelination, ultimately revealing new therapeutic targets for multiple sclerosis (MS) and related neurodegenerative conditions such as Alzheimer's disease.

The work is grounded in the understanding that PKC is a key downstream effector of TREM2 and PLC gamma-2, two molecules strongly implicated in neuroinflammatory disease pathogenesis. Unraveling bryostatin-1-induced post-translational modifications will provide mechanistic insight into how innate immune cells can be therapeutically reprogrammed to support CNS repair.

Technical specifications

Research approach:

• In vitro phosphoproteomic analysis: Cultured microglia will be treated with bryostatin-1 or vehicle and challenged with pro-inflammatory LPS, followed by mass spectrometry-based comparison of phosphorylated protein profiles
• In vivo phosphoproteomic analysis: The well-established EAE (experimental autoimmune encephalomyelitis) mouse model of inflammatory MS will be used to compare CNS innate immune phosphoproteomes between bryostatin-1-treated and vehicle-treated groups
• Pathway mapping: Identified phosphoproteins will be mapped to signaling cascades downstream of PKC, TREM2, and PLC gamma-2 to pinpoint key regulatory nodes in neuroinflammation and remyelination

Key features:

• Leverages bryostatin-1, a clinically relevant compound with demonstrated brain penetrance and immunomodulatory activity
• Applies state-of-the-art phosphoproteomics to characterize post-translational modifications in CNS innate immune cells
• Bridges molecular profiling with disease-relevant functional outcomes in an established MS animal model
• Expected project duration: approximately one year

Technology readiness level

This research is at an early discovery stage, building on validated preliminary findings that bryostatin-1 activates anti-inflammatory and regenerative transcriptional programs in microglia and CNS macrophages. The proposed phosphoproteomic profiling represents a foundational mechanistic study that will generate new knowledge about PKC-mediated signaling in CNS innate immunity. Results are expected to identify and prioritize novel therapeutic targets for MS and related neurodegenerative diseases, creating a basis for subsequent drug discovery and development efforts. The team is seeking partnership support to conduct this one-year investigation.