Characterization of RNA Cargo from Microglial Extracellular Vesicles in ALS and FTD
Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are progressive neurodegenerative diseases caused by various gene mutations. The goal of this study will be to investigate changes in the RNA cargo of microglia-derived extracellular vesicles (MDEVs) from subjects with frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), and normal controls. Extracellular vesicles (EVs) are released from all cell types and carry molecular cargo that reflects the activities of the origin cell. Previous studies have shown that EVs play a role in transferring cellular cargo such as RNA, DNA, and proteins to target cells. After delivery, the vesicles have the potential to modify the target cell’s phenotype and advance the progression of various diseases.
Microglia are the primary immune cells in the central nervous system and function as tissue-resident macrophages. They can become activated by pathological events and may even contribute to the spread of pathology in neurodegenerative diseases. For this study, human-induced pluripotent stem cells (iPSCs) were generated from patient samples and differentiated into microglia. Cell culture media from these microglia cultures were collected and EVs were isolated using size exclusion chromatography (SEC). Before we began the studies on the microglia media, we tested our EV collection methods using two SEC columns, one packed with 70 nm beads and one with 35 nm beads. We present our results of EV numbers, sizes, RNA concentration and proteins from our tests.