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Randolph, C. E.

Publications and source records attributed to Randolph, C. E..

5 recordsLinked to original sources

Sleep-dependent clearance of brain lipids by peripheral blood cells

Sleep is typically viewed through a brain-centric lens, with little known about the role of the periphery. Here, we identify a sleep function for peripheral macrophage-like cells (hemocytes) in the Drosophila circulation, showing that hemocytes track to the brain during sleep and take up lipids accumulated in cortex glia due to wake-associated oxidative damage. Through a screen of phagocytic receptors expressed in hemocytes, we discovered that knockdown of eater, a member of the Nimrod receptor family, reduces sleep. Loss of eater also disrupts hemocyte adhesion to the brain and lipid uptake, which results in increased brain levels of Acetyl CoA and acetylated proteins, including mitochondrial proteins PGC1 and DRP1. Dysregulation of mitochondria, reflected in high oxidation and reduced NAD+, is accompanied by impaired memory and lifespan. Thus, peripheral blood cells, which we suggest are precursors of mammalian microglia, perform a daily function of sleep to maintain brain function and fitness.

molecular biology↗

Impact Induces Phagocytic Defect in Reactive Microglia

We have developed traumatic brain injury (TBI)-on-a-chip in vitro models using primary microglia and neuronal networks and recorded the molecular and cellular changes following impact to represent impact injury. Using a pH-responsive amyloid {beta} (A{beta}pH), we showed that microglial phagocytosis was reduced at 7 days post-impact on the chip. Simultaneously, neurons increased their uptake of A{beta}, and decreased neuronal firing frequency at 7 days post-impact based on electrophysiological recordings. Given the importance of lipid metabolism in brain trauma and neurodegeneration, the lipidome secreted by impacted cells was analyzed to understand changes in cellular processes. Interestingly, many lipid species from the sphingomyelin, glycerophospholipid, and phosphatidylserine classes were significantly affected by impact, which are known to play important roles in the resolution of neuroinflammation and the pathogenesis of neurodegeneration.

neuroscience↗

Neutral lipid processing in glia is sexually dimorphic and promotes sleep through diacylglycerol catabolism

Sleep is thought to have a protective role in clearing toxic waste from the brain, which may include processing of damaged lipids. We recently showed that blocking endocytosis in glia increases sleep and report here that this block is associated with an increase in peroxidized lipids and glial lipid droplet accumulation, raising the possibility that accumulation of these lipids increases the need to sleep. Sleep gain induced by blocking glial transport is exaggerated by knockout of the lipid droplet coat protein, Lipid Storage Droplet 2 (Lsd2), suggesting that sleep-promoting lipids are not contained in lipid droplets. To identify lipids regulated by sleep state, we performed global, targeted lipidomics analysis on Drosophila neurons and glia, screening nearly 3,000 lipids across 11 major classes. This revealed that sex influences lipid composition in both cell types and lipid homeostasis following extended wakefulness. Female neurons and glia are enriched in ultra-long chain fatty acids, triacylglycerols, and diacylglycerols, with glial diacylglycerol enrichment correlating with elevated sleep need. Based on manipulations of neutral lipid metabolic pathways, we propose that monoacylglycerols, products of glial diacylglycerol catabolism, promote sleep.

neuroscience↗

Lesion-remote astrocytes govern microglia-mediated white matter repair

Spared regions of the damaged central nervous system undergo dynamic remodeling and exhibit a remarkable potential for therapeutic exploitation. Here, lesion-remote astrocytes (LRAs), which interact with viable neurons, glia and neural circuitry, undergo reactive transformations whose molecular and functional properties are poorly understood. Using multiple transcriptional profiling methods, we interrogated LRAs from spared regions of mouse spinal cord following traumatic spinal cord injury (SCI). We show that LRAs acquire a spectrum of molecularly distinct, neuroanatomically restricted reactivity states that evolve after SCI. We identify transcriptionally unique reactive LRAs in degenerating white matter that direct the specification and function of local microglia that clear lipid-rich myelin debris to promote tissue repair. Fueling this LRA functional adaptation is Ccn1, which encodes for a secreted matricellular protein. Loss of astrocyte CCN1 leads to excessive, aberrant activation of local microglia with (i) abnormal molecular specification, (ii) dysfunctional myelin debris processing, and (iii) impaired lipid metabolism, culminating in blunted debris clearance and attenuated neurological recovery from SCI. Ccn1-expressing white matter astrocytes are specifically induced by local myelin damage and generated in diverse demyelinating disorders in mouse and human, pointing to their fundamental, evolutionarily conserved role in white matter repair. Our findings show that LRAs assume regionally divergent reactivity states with functional adaptations that are induced by local context-specific triggers and influence disorder outcome. Astrocytes tile the central nervous system (CNS) where they serve vital roles that uphold healthy nervous system function, including regulation of synapse development, buffering of neurotransmitters and ions, and provision of metabolic substrates1. In response to diverse CNS insults, astrocytes exhibit disorder-context specific transformations that are collectively referred to as reactivity2-5. The characteristics of regionally and molecularly distinct reactivity states are incompletely understood. The mechanisms through which distinct reactivity states arise, how they evolve or resolve over time, and their consequences for local cell function and CNS disorder progression remain enigmatic. Immediately adjacent to CNS lesions, border-forming astrocytes (BFAs) undergo transcriptional reprogramming and proliferation to form a neuroprotective barrier that restricts inflammation and supports axon regeneration6-9. Beyond the lesion, spared but dynamic regions of the injured CNS exhibit varying degrees of synaptic circuit remodeling and progressive cellular responses to secondary damage that have profound consequences for neural repair and recovery10,11. Throughout these cytoarchitecturally intact, but injury-reactive regions, lesion-remote astrocytes (LRAs) intermingle with neurons and glia, undergo little to no proliferation, and exhibit varying degrees of cellular hypertrophy7,12,13. The molecular and functional properties of LRAs remain grossly undefined. Therapeutically harnessing spared regions of the injured CNS will require a clearer understanding of the accompanying cellular and molecular landscape. Here, we leveraged integrative transcriptional profiling methodologies to identify multiple spatiotemporally resolved, molecularly distinct states of LRA reactivity within the injured spinal cord. Computational modeling of LRA-mediated heterotypic cell interactions, astrocyte-specific conditional gene deletion, and multiple mouse models of acute and chronic CNS white matter degeneration were used to interrogate a newly identified white matter degeneration-reactive astrocyte subtype. We define how this reactivity state is induced and its role in governing the molecular and functional specification of local microglia that clear myelin debris from the degenerating white matter to promote repair.

neuroscience↗

Amyloid β Induces Lipid Droplet-Mediated Microglial Dysfunction in Alzheimer's Disease

Several microglia-expressed genes have emerged as top risk variants for Alzheimers disease (AD). Impaired microglial phagocytosis is one of the main proposed outcomes by which these AD-risk genes may contribute to neurodegeneration, but the mechanisms translating genetic association to cellular dysfunction remain unknown. Here we show that microglia form lipid droplets (LDs) upon exposure to amyloid-beta (A{beta}), and that their LD load increases with proximity to amyloid plaques in brains from human patients and the AD mouse model 5xFAD. LD formation is dependent on age and disease progression and is prominent in the hippocampus in mice and humans. Despite differences in microglial LD load between brain regions and sexes in mice, LD-laden microglia exhibited a deficit in A{beta} phagocytosis. Unbiased lipidomic analysis identified a decrease in free fatty acids (FFAs) and a parallel increase in triacylglycerols (TGs) as the key metabolic transition underlying LD formation. DGAT2, a key enzyme for converting FFAs to TGs, promotes microglial LD formation and is increased in 5xFAD and human AD brains. Inhibition or degradation of DGAT2 improved microglial uptake of A{beta} and drastically reduced plaque load in 5xFAD mice, respectively. These findings identify a new lipid-mediated mechanism underlying microglial dysfunction that could become a novel therapeutic target for AD.

neuroscience↗