✍🏼 Reilly L Allison , Allison D Ebert

🏠 Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, WI , United States

📑 Human Molecular Genetics (2024)

Abstract
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the loss of upper and lower motor neurons (MNs). The loss of MNs in ALS leads to muscle weakness and wasting, respiratory failure, and death often within two years of diagnosis. Glial cells in ALS show aberrant expression of pro-inflammatory and neurotoxic proteins associated with activation and have been proposed as ideal therapeutic targets. In this study, we examined astrocyte-targeted treatments to reduce glial activation and neuron pathology using cells differentiated from ALS patient-derived iPSC carrying SOD1 and C9ORF72 mutations. Specifically, we tested the ability of increasing interleukin 10 (IL-10) and reducing C-C motif chemokine ligand 2 (CCL2/MCP-1) signaling targeted to astrocytes to reduce activation phenotypes in both astrocytes and microglia. Overall, we found IL10/CCL2NAb treated astrocytes to support anti-inflammatory phenotypes and reduce neurotoxicity, through different mechanisms in SOD1 and C9ORF72 cultures. We also found altered responses of microglia and motor neurons to astrocytic influences when cells were cultured together rather than in isolation. Together these data support IL-10 and CCL2 as non-mutation-specific therapeutic targets for ALS and highlight the role of glial-mediated pathology in this disease.

How the WOLF was used in this study
Researchers used a WOLF Cell Sorter to purify and expand stable fluorescent iPSC lines. After creating GFP- and RFP-expressing induced pluripotent stem cell (iPSC) lines from ALS patient and control backgrounds via lentiviral transduction, the authors isolated GFP⁺ and RFP⁺ cells using the WOLF Cell Sorter to obtain highly purified populations expressing these fluorescent markers for downstream applications. This sorting step was critical for generating homogeneous, stably labeled microglia and motor neuron populations that could be reliably used in subsequent functional assays, co-cultures, and imaging experiments to assess how astrocyte-targeted modulation of IL-10 and CCL2 impacts glial and neuronal phenotypes in ALS models.