bims-microg Biomed News
on Microglia in health and disease
Issue of 2025–11–16
twenty-one papers selected by
Marcus Karlstetter, Universität zu Köln



  1. Int J Mol Sci. 2025 Oct 31. pii: 10630. [Epub ahead of print]26(21):
      Diabetic retinopathy (DR), a major complication of diabetes, is driven by chronic inflammation in which retinal microglial cells play a central role. The Hippo pathway kinases NDR1/2 regulate macrophage function, but their role in microglia and DR remain unknown. This study investigates the function of the NDR2 kinase in microglial cells under high-glucose (HG) conditions. Using CRISPR-Cas9, we partially knocked out the Ndr2/Stk38l gene in BV-2 mouse microglial cells and analyzed metabolic activity, phagocytosis, migration, and cytokine release. We confirmed NDR2 expression in microglia and observed increased levels under HG, suggesting a role in hyperglycemia-induced stress. Ndr2/Stk38l (hereafter referred to as Ndr2) downregulation impaired mitochondrial respiration and reduced metabolic flexibility, indicating defective stress adaptation. Functionally, microglia with a partial downregulation of Ndr2 displayed reduced phagocytic and migratory capacity-both dependent on cytoskeletal dynamics. Moreover, Ndr2 downregulation altered the secretory profile, elevating pro-inflammatory cytokines (IL-6, TNF, IL-17, IL-12p70) even under normal glucose levels. These findings identify NDR2 protein kinase as a key regulator of microglial metabolism and inflammatory behavior under diabetic conditions. By modulating immune and metabolic responses, NDR2 may contribute to the neuroinflammatory processes underlying DR. Targeting NDR2 function in microglia may offer novel therapeutic strategies to mitigate retinal inflammation and progression of DR.
    Keywords:  Ndr2/Stk38l gene; Nuclear Dbf2-Related (NDR) kinases; diabetic retinopathy; high-glucose; inflammation; metabolism; microglia; retina
    DOI:  https://doi.org/10.3390/ijms262110630
  2. Science. 2025 Nov 13. 390(6774): eadq1177
      Multiple sclerosis (MS) progresses through brain region-specific inflammation and degeneration, with poorly defined mechanisms. In individuals with MS, we identified increased expression of formyl peptide receptor 1 (FPR1) in central nervous system (CNS)-resident microglia and CNS-infiltrating macrophages. Blood amounts of N-formylated peptides, which are endogenous agonists of FPR1, correlated with disease progression in patients with MS. In MS mouse models, signaling through FPR1 promoted microglial mitochondrial dysfunction, causing axonal loss and apoptosis. FPR1-expressing microglia sustained the clonal expansion of myelin-reactive CD4+ T cells in the CNS. A CNS-penetrating small molecule FPR1 antagonist, T0080, mitigated autoimmune responses and axonal degeneration. Our study identifies FPR1 signaling as a potential mechanism for MS progression and suggests antagonizing FPR1 as a therapeutic approach.
    DOI:  https://doi.org/10.1126/science.adq1177
  3. Nat Commun. 2025 Nov 12. 16(1): 9951
      Multiple sclerosis (MS) is a chronic, inflammatory condition characterized by neurodegeneration and lost myelin, or demyelination. This lost myelin may be regenerated in people with MS through a process called remyelination, that is prone to failure and is impaired with age. Remyelination is facilitated by microglia but our understanding of the microglial response during remyelination is incomplete. Here, we profile the microglial response during remyelination in the lysolecithin mouse model using single-cell RNA sequencing and find several distinct microglial states during the early stages of remyelination that coalesce into a resolved state defined by the presence of myelin transcripts, a state also present in MS brains. We also observe a delay in the appearance of several microglial states with age, in concordance with delayed remyelination. This multi-faceted microglial response during efficient remyelination provides the basis of multi-faceted microglia-specific targets for future MS therapies.
    DOI:  https://doi.org/10.1038/s41467-025-64906-w
  4. Adv Sci (Weinh). 2025 Nov 11. e08957
      Depression is one of the most prevalent neuropsychiatric disorders worldwide, and multiple studies have implicated metabolic dysfunction in its pathophysiology. However, the molecular mechanisms by which metabolic pathways modulate depressive‑like behavior remain largely uncharacterized. Here, this work finds that the CDGSH iron sulfur domain 1 (CISD1), a redox protein localized to the outer mitochondrial membrane, is upregulated in the medial prefrontal cortex after chronic stress. Pharmacological inhibition and genetic knockdown of CISD1 significantly ameliorate depressive-like behavior in mice, and CISD1 knockdown also reverse microglial inflammatory activation. Moreover, this work finds that chronic stress specifically upregulates microglial CISD1 expression, and that conditional knockout of microglial CISD1 alleviates neuroinflammation and depressive‑like behavior in mice. Mechanistically, chronic stress promotes NADH oxidation to generate NAD⁺ by upregulating CISD1 expression. The elevated NAD⁺ functions as a cofactor for glyceraldehyde-3-phosphate dehydrogenase, accelerating glycolysis and promoting inflammatory activation. Pioglitazone exerts antidepressant effects by inhibiting NADH oxidation through a CISD1-dependent pathway in microglia. In conclusion, this study elucidates the role of CISD1 in microglial metabolism, establishing a robust experimental foundation for screening potential antidepressant drugs.
    Keywords:  CDGSH iron sulfur domain 1; depression; medial prefrontal cortex; microglial inflammatory activation; pioglitazone
    DOI:  https://doi.org/10.1002/advs.202508957
  5. Brain Behav Immun. 2025 Nov 11. pii: S0889-1591(25)00420-9. [Epub ahead of print] 106178
      Traumatic brain injury (TBI) affects over 69 million people every year, and mild traumatic brain injury (mTBI) accounts for 70-90 % of cases. TBI has two components: i) primary injury - direct damage to the tissue from the mechanical impact and ii) secondary injury - additional or extended damage to the tissue from the ensuing biochemical and physiological processes such as neuroinflammation. Neuroinflammation triggered in part by activated microglia, determines whether the post-injury outcome is recovery or long-term neurodegeneration. Microglia, key components of the neuroinflammatory process, release cytokines such as TNF-α, which affect neuronal activity. Our study investigated the effects of acute microglia depletion on hippocampal neurophysiology in male mice (7-10 days after mTBI), a time window that allows us to target sub-acute microglial responses post-injury. An additional objective of the study was to determine if the pro-inflammatory cytokine TNF-α contributed to the injury-induced network excitability shifts in the hippocampal circuitry. We demonstrate that depleting microglia with PLX-3397 treatment for 7-10 days after mTBI restores network excitability in hippocampal area CA1 and the dentate gyrus (DG). Furthermore, treatments with thalidomide and etanercept show that TNF-α plays a role in altering the network excitability after mTBI. These findings provide new insights into the physiological changes after injury and highlight potential targets for future interventions to specifically address the detrimental effects of chronic inflammation.
    Keywords:  Circuit function; Contextual fear conditioning; Electrophysiology; Extinction; Hippocampus; Microglia depletion; Neuroinflammation; mTBI
    DOI:  https://doi.org/10.1016/j.bbi.2025.106178
  6. Brain Behav Immun. 2025 Nov 12. pii: S0889-1591(25)00422-2. [Epub ahead of print] 106180
      Both clinical and preclinical evidence demonstrates a robust association between mood disorders and cerebrovascular diseases, but the underlying mechanism remains elusive. In this study, to model the comorbidity of depression and cerebral hypoperfusion, we combined two different types of chronic stress paradigms, chronic unpredictable mild stress (CUMS) and social defeat (SD), with bilateral carotid artery stenosis (BCAS). Mice in the comorbidity group exhibited additive impairments in cognitive behaviors, surpassing the effects observed in the sham, stress-only, or BCAS-only groups. Notably, the most prominent change was massive microglial activation in the comorbidity group extending into critical gray matter areas, accompanied by severe brain lesion including blood-brain barrier (BBB) damage, demyelination, localized neuronal disruption, and abnormal vessel formation. Importantly, microglia emerged as central players in all observed cellular events, displayed stage-specific roles mediated by distinct subpopulations, driving neuroinflammation in response to BCAS and promoting angiogenesis under comorbid condition. Our findings suggest chronic stress impairs microglial function, increasing vulnerability to cerebral hypoperfusion. This study highlights chronic stress as a key risk factor for cerebrovascular events and underscores the importance of stress management in patients with this comorbidity.
    Keywords:  Carotid stenosis; Chronic stress; Depression; Ischemia; Microglia
    DOI:  https://doi.org/10.1016/j.bbi.2025.106180
  7. Adv Sci (Weinh). 2025 Nov 12. e13270
      Glaucoma is a leading cause of irreversible blindness, characterized by progressive retinal ganglion cells (RGCs) loss. Increasing evidence links microglial activation and lipid metabolism dysregulation to neurodegeneration. However, the role of microglial lipid metabolic reprogramming in disease pathogenesis remains unclear. This study finds that microglia in an acute ocular hypertension (AOH) model exhibit abnormal lipid droplet accumulation, downregulation of low-density lipoprotein receptor-related protein 1 (LRP1), and a shift toward a pro-inflammatory M1 phenotype. Importantly, serum samples from glaucoma patients reveal significantly reduced LRP1 levels compared to controls. To restore lipid homeostasis, this study develops alpinetin-loaded PLGA nanoparticles (AlpNPs), which demonstrate efficient microglial uptake and sustained release. AlpNPs reduced intracellular lipid accumulation, promoted M2 polarization, and suppressed microglial proliferation and migration. Mechanistically, AlpNPs directly bind to LRP1 and enhance its interaction with PPARγ, thereby activating the downstream LXRα-ABCA1 pathway, which is pivotal for cholesterol efflux and anti-inflammatory responses. Knockdown of LRP1 abolished the protective effects of AlpNPs, confirming its essential role in mediating metabolic reprogramming. In vivo, intravitreal injection of AlpNPs significantly attenuates retinal inflammation and preserves RGCs in AOH. These findings identify microglial lipid metabolic dysfunction as a key driver in glaucoma and highlight LRP1-targeted nanotherapy as a promising strategy for neuroprotection.
    Keywords:  AlpNPs; LRP1‐PPARγ; glaucoma; lipid metabolism; microglia polarization
    DOI:  https://doi.org/10.1002/advs.202513270
  8. Adv Sci (Weinh). 2025 Nov 12. e16269
      Glial scar formation represents a significant obstacle to neural regeneration following spinal cord injury (SCI), evolving from a protective glial response in the acute phase to a fibrotic and inhibitory barrier in the chronic stage. In this study, chondroitin sulphate proteoglycans (CSPGs) are identified as key regulators of scar maturation via a pathogenic microglia-astrocyte axis. CSPGs promote the transition of reactive astrocytes (RAs) into scar-forming astrocytes (SAs) by inducing a pro-inflammatory microglial phenotype. Mechanistically, CSPGs suppress cytochrome P450 (CYP450) enzyme activity in microglia, disrupting metabolic homeostasis and perpetuating inflammatory responses. Targeted degradation of CSPGs reprogrammes microglia toward an anti-inflammatory state, thereby attenuating SA differentiation and fibrotic matrix deposition. To enable spatiotemporally precise intervention, a reactive oxygen species-responsive, connective tissue growth factor-binding fusogenic lipopolyplex for RA-targeted delivery of the chondroitinase ABC (ChABC) gene is designed. This platform selectively degrades CSPGs at the lesion border, interrupts the maladaptive glial feedback loop, and facilitates scar-free repair after SCI. These findings reveal a metabolic mechanism underlying glial scarring and propose a precision nanotherapeutic strategy to modulate the SCI microenvironment, thereby enhancing neuronal regeneration and functional recovery.
    Keywords:  chondroitin sulphate proteoglycan degradation; microglia–astrocyte crosstalk; scar‐free repair; spinal cord injury; targeted gene delivery; tissue regeneration
    DOI:  https://doi.org/10.1002/advs.202516269
  9. Metab Brain Dis. 2025 Nov 15. 40(8): 313
      Vulnerable brain models exhibit heightened susceptibility to anesthetic neurotoxicity, with diabetic offspring warranting particular attention, yet the underlying mechanisms remain unclear. This study integrated bioinformatics and experiments to investigate the molecular basis of sevoflurane-induced neurodevelopmental toxicity in diabetic offspring. Offspring of streptozotocin-induced diabetic dams (gestational diabetes model) and control mice were exposed to sevoflurane (2.5%, 3 × 2 h, P6-P8). Hippocampal tissues underwent RNA sequencing. Bioinformatics analysis identified dysregulated pathways, followed by validation of neuronal apoptosis (TUNEL), synaptic proteins (PSD-95, Synaptophysin), and microglial activity (Iba1+). Hub genes were screened via protein-protein interaction networks. Diabetic-sevoflurane offspring showed aberrant activation of neural crest differentiation, oxidative stress/redox pathways, and microglial pathogen phagocytosis. Compare with the Control group the diabetic-sevoflurane offspring showed increased apoptosis, synaptic loss and enhanced Iba1 + microglial density. Uty, Uba1y, Ddx3y, Kdm5d, and Eif2s3y were identified as key regulators. Gestational diabetes primes microglia via Hub gene networks, amplifying sevoflurane-induced neurotoxicity. Targeting these genes may mitigate risks in diabetic offspring requiring anesthesia.Clinical trial number: Not applicable.
    Keywords:  Anesthetic; Gestational diabetes; Microglia; Neuron; Neurotoxicity
    DOI:  https://doi.org/10.1007/s11011-025-01746-z
  10. Acta Neuropathol Commun. 2025 Nov 14. 13(1): 234
      Exposure to pesticides, such as rotenone or paraquat, is an environmental factor that plays an important role in the pathogenesis of Parkinson's disease (PD). Rotenone induces PD-like pathology and is therefore used to develop parkinsonian animal models. Dopaminergic neurotoxicity caused by rotenone has been attributed to the inhibition of mitochondrial complex I, oxidative stress and neuroinflammation; however, the mechanisms underlying selective dopaminergic neurodegeneration by rotenone remain unclear. To resolve this, we focused on glial diversity and examined whether the brain region-specific glial response to rotenone could determine the vulnerability of dopaminergic neurons using primary cultured neurons, astrocytes and microglia from the midbrain and striatum of rat embryos and rotenone-injected PD model mice. Direct neuronal treatment with low-dose rotenone failed to damage dopaminergic neurons. Conversely, rotenone exposure in the presence of midbrain astrocyte and microglia or conditioned media from rotenone-treated midbrain glial cultures containing astrocytes and microglia produced dopaminergic neurotoxicity, but striatal glia did not. Surprisingly, conditioned media from rotenone-treated midbrain astrocytes or microglia monocultures did not affect neuronal survival. We also demonstrated that rotenone targeted midbrain astrocytes prior to microglia to induce dopaminergic neurotoxicity. Rotenone-treated astrocytes produced secreted protein acidic and rich in cysteine (SPARC) extracellularly, which induced microglial proliferation, increase in IL-1β and TNF-α, and NF-κB (p65) nuclear translocation in microglia, resulting in dopaminergic neurodegeneration. In addition, rotenone exposure caused the secretion of NFAT-related inflammatory cytokines and a reduction in the level of an antioxidant metallothionein (MT)-1 from midbrain glia. Furthermore, we observed microglial proliferation and a decrease in the number of MT-positive astrocytes in the substantia nigra, but not the striatum, of low-dose rotenone-injected PD model mice. Our data highlight that rotenone targets midbrain astrocytes, leading to SPARC secretion, which promotes the neurotoxic conversion of microglia and leads to glial dysfunction-mediated dopaminergic neurodegeneration.
    Keywords:  Astrocyte; Microglia; Parkinson's disease; Rotenone; SPARC
    DOI:  https://doi.org/10.1186/s40478-025-02160-3
  11. Cell Rep. 2025 Nov 08. pii: S2211-1247(25)01315-4. [Epub ahead of print]44(11): 116544
      Retinal microglia originate during embryonic hematopoiesis and play an essential role in visual function. However, the mechanisms guiding microglial precursors into the retina remain poorly understood. Here, we show that, in zebrafish embryos, microglial precursors enter the vitreous space via blood vessels, mainly the ventral radial vessel/hyaloid vein, before migrating to the developing retina. Vascular Lamb1 provides a haptotactic gradient sensed by Itga6 in microglial precursors, which activates Rac1 signaling to promote F-actin polarization and directional migration. Disruption of blood vessels or interference with the Lamb1-Itga6 axis blocks microglial precursor entry into the vitreous space and reduces retinal colonization. Similarly, LAMB1 is deposited in the blood vessels of mice, and endothelial-specific deficiency impairs microglial precursor settlement. These findings indicate a conserved Lamb1-Itga6 signaling axis that guides retinal microglial migration during development.
    Keywords:  CP: cell biology; CP: neuroscience; Itga6; Lamb1; blood vessel; microglial precursor; retina
    DOI:  https://doi.org/10.1016/j.celrep.2025.116544
  12. Eur J Neurosci. 2025 Nov;62(9): e70281
      Microglia, the brain-resident macrophages, are critically involved in numerous physiological and pathological brain processes, including neoplasms, epilepsy, and neurodegeneration. However, investigating microglial function is notoriously difficult because they are extremely sensitive to changes in their environment and drastically alter their transcriptional state and morphology once they are isolated from the brain and cultured in vitro. In vivo experiments in mice are likewise limited because of vast differences between mouse and human microglia, particularly regarding the expression of disease-associated genes. To overcome this issue, we developed a highly controlled in vitro cerebral organoid platform where human microglia adopt an in vivo-like phenotype. This approach allows long-term studies and high-throughput analysis of in vivo-like human microglia suitable for disease modeling and drug testing.
    Keywords:  RNA‐seq; cerebral organoids; human stem cells; microglia
    DOI:  https://doi.org/10.1111/ejn.70281
  13. Invest Ophthalmol Vis Sci. 2025 Nov 03. 66(14): 26
       Purpose: Retinoblastoma (RB) is the most prevalent intraocular malignancy in children, which significantly impacts patients' quality of life. Chimeric antigen receptor (CAR) immune cell therapies opened a door to treating tumors. However, whether microglia, the resident immune cell in the retina, could be engineered to treat retinoblastoma remains unknown. The purpose of this study is to generate human CAR-microglia and to investigate the antitumor effects of CAR-microglia in RB.
    Methods: The presence of CAR in microglia cells was verified using a fluorescence microscope, PCR, and flow cytometry analysis. The CAR-microglia generated from induced pluripotent stem cells were identified by immunofluorescence, flow cytometry analysis, and RNA sequencing (RNA-seq). The antitumor effects of CAR-microglia in retinoblastoma are investigated in vitro and in vivo by live imaging, flow cytometry analysis, RNA-seq, quantitative real-time PCR, cell viability analysis, cell bioluminescence analysis, ELISA, optical coherence tomography, fundus photography, bioluminescence imaging, and hematoxylin and eosin staining.
    Results: We developed human GD2 CAR-microglia and found that they exhibited a remarkable phagocytic effect against retinoblastoma Y79 cells and retinoblastoma organoids, as demonstrated by live imaging. Moreover, GD2 CAR-microglia administration to immunodeficient mice carrying a retinoblastoma xenograft resulted in a significant reduction in tumor growth and prolonged survival.
    Conclusions: Our findings demonstrate a potent antitumor effect of GD2-CAR-engineered microglia in retinoblastoma, offering compelling evidence to support the continued development of CAR-microglia as a therapeutic strategy for this disease.
    DOI:  https://doi.org/10.1167/iovs.66.14.26
  14. Alzheimers Dement. 2025 Nov;21(11): e70879
       INTRODUCTION: Lipid regulation is crucial role in Alzheimer's disease (AD) pathogenesis. In AD, microglia show elevated sterol O-acyltransferase 1/Acyl-coenzymeA: Choleseterol Acyltransferase 1 (SOAT1) expression, encoding Acyl-coenzymeA: Cholesterol Acyltransferase 1  (ACAT1), which produces cholesteryl esters (CEs) in lipid droplets. Inhibiting ACAT1 has been shown to reduce amyloid beta (Aβ) pathology, though the mechanism is unclear.
    METHODS: We inhibited ACAT1 using avasimibe (AV) in wild-type, triggering receptor expressed on myeloid cells 2 (TREM2) knockout (KO), and low-density lipoprotein receptor related protein 1 (LRP1) KO mouse BV2 and human induced pluripotent stem cell-derived microglia and measured the impact on Aβ uptake to determine the mechanism through which the inhibition of ACAT1 enhances Aβ uptake.
    RESULTS: ACAT1 inhibition increased LRP1 levels and soluble TREM2 (sTREM2) release via enhanced TREM2 cleavage by ADAM metallopeptidase domain 10/17 (ADAM10/17). KO of TREM2 or blockade of sTREM2 release prevented AV-enhanced Aβ uptake. This effect was rescued by recombinant sTREM2, but only when LRP1 was present.
    DISCUSSION: ACAT1 inhibition promotes microglial Aβ uptake in a sTREM2- and LRP1-dependent manner, offering insights into novel therapeutic strategies for AD.
    HIGHLIGHTS: Inhibition of ACAT1, the major enzyme that catalyzes cholesterol storage via esterification enhances microglia-mediated Aβ uptake. Increased Aβ uptake is dependent on the presence of both TREM2 and LRP1. Inhibition of ACAT1 increases cleavage of TREM2 via ADAM10/17 to release sTREM2. Treatment of microglial cells with sTREM2 rescues Aβ uptake in TREM2 KO BV2 cells. Inhibition of ACAT1 promotes Aβ uptake through increased shedding of TREM2, which enhances Aβ uptake through a LRP1-dependent mechanism.
    Keywords:  ACAT1/SOAT1; Alzheimer's disease; LRP1; TREM2; amyloid beta; lipid droplets; microglia; phagocytosis
    DOI:  https://doi.org/10.1002/alz.70879
  15. Mol Ther. 2025 Nov 12. pii: S1525-0016(25)00872-X. [Epub ahead of print]
      Recent advances in cell-based therapies have demonstrated therapeutic safety and efficacy for a variety of diseases. However, significant challenges remain in their development for the treatment of neurological disorders. Preclinical studies have explored microglial replacement strategies utilizing bone marrow transplantation and CSF1R inhibitor (CSF1Ri) treatment. However, these approaches often require highly invasive strategies and result in engrafted cells that remain transcriptionally and functionally distinct from microglia. To assess less-invasive microglia replacement strategies capable of preserving microglial ontogeny, we developed a Csf1r-G793A knockin mouse. We found that G793A mice develop typical microglial densities and peripheral hematopoietic populations, which exhibit broad CSF1Ri resistance enabling widespread microglia replacement following direct intraparenchymal injection or bone marrow transplantation. Furthermore, we demonstrate that widespread microglia replacement can be achieved via less-invasive intracisternal delivery of CSF1Ri-resistant murine and induced pluripotent stem cell (iPSC)-derived human microglia. Together with the accompanying study by Lombroso and colleagues, our findings demonstrate that G793A mice provide a robust and broadly applicable source of engraftable donor microglia and peripheral macrophages for the preclinical investigation of microglia replacement strategies.
    Keywords:  CSF1R inhibitors; cell therapies; iPSC-microglia; immune cell therapy; microglia; microglia replacement; microglia transplantation; routes of delivery
    DOI:  https://doi.org/10.1016/j.ymthe.2025.10.051
  16. Alcohol. 2025 Nov 06. pii: S0741-8329(25)00125-9. [Epub ahead of print]
      Alcohol use disorders (AUDs) affect substantial populations worldwide and increase the risk of developing cognitive impairments and alcohol-associated dementia. While chronic inflammatory signaling likely plays an important role in alcohol-associated neurological sequalae, the precise mechanisms underlying alcohol-associated neuropathology remain enigmatic. We hypothesize that alcohol leads to neuroimmune dysregulation among neurons, astrocytes, and microglia; and is perpetuated by innate immune signaling pathways involving cell-cell signaling. To investigate how alcohol dysregulates neuroimmune interactions in a human context, we constructed a triculture model comprising neurons, astrocytes, and microglia derived from human induced pluripotent stem cells. After exposure to ethanol, we observed significant differential gene expression relating to innate immune pathways, inflammation, and microglial activation. Microglial activation was confirmed with morphological analysis and expression of CD68, a lysosomal-associated membrane protein and marker for phagocytic microglial activation. A striking finding in our study was the elevation of TREM2 expression and, specifically, TREM2 alternatively spliced isoforms that are predicted to give rise to soluble TREM2. TREM2 loss-of-function variants have been reported to be a risk factor for Alzheimer's disease. These results suggest that ethanol exposure in the brain may lead to increased microglial activation and production of soluble isoform named TREM2219 through alternate splicing. Deciphering the molecular and cellular mechanisms underpinning ethanol-related neuroimmune dysregulation within a human context promises to shed light on the etiology of AUD-related disorders, potentially contributing to the development of effective therapeutic strategies.
    Keywords:  AUD; Microglia; TREM2; iPSC; neuron-glia triculture
    DOI:  https://doi.org/10.1016/j.alcohol.2025.10.006
  17. Free Radic Biol Med. 2025 Nov 11. pii: S0891-5849(25)01353-X. [Epub ahead of print]
      Retinopathy of prematurity (ROP) is a leading cause of childhood blindness and is characterized by retinal neovascularization. Microglia have emerged as key mediators of hypoxia-induced angiogenesis, yet the underlying molecular mechanisms remain incompletely defined. Heme oxygenase-1 (HMOX1) is a stress-inducible enzyme with well-established antioxidant and anti-inflammatory properties, acting as an important electrophilic signaling mediator in redox biology. Here, we investigated the role of microglial HMOX1 in retinal angiogenesis and its regulation of endothelial signaling. Using an oxygen-induced retinopathy (OIR) mouse model combined with bulk and single-cell RNA sequencing, we identified HMOX1 as a hypoxia-induced angiogenic regulator selectively enriched in retinal microglia. Conditional knockout of HMOX1 in microglia markedly reduced neovascularization in vivo, while in vitro hypoxia-driven microglia-endothelial co-culture assays confirmed impaired endothelial proliferation, migration, and tube formation. Mechanistically, loss or pharmacological inhibition of HMOX1 suppressed STAT3 activation and VEGF expression in endothelial cells, establishing a paracrine signaling pathway between microglia and endothelium. These findings demonstrate that microglial HMOX1 drives pathological angiogenesis by activating the endothelial STAT3-VEGF axis. Our study uncovers a novel hypoxia-microglia-endothelium signaling mechanism, highlighting microglial HMOX1 as a promising therapeutic target for ROP and other hypoxia-related angiogenic diseases.
    Keywords:  Endothelial cells; Heme oxygenase-1; Microglia; Oxygen-induced retinopathy; Retinal angiogenesis; STAT3
    DOI:  https://doi.org/10.1016/j.freeradbiomed.2025.11.018
  18. Cells. 2025 Oct 28. pii: 1687. [Epub ahead of print]14(21):
      Human microglia are central regulators and actors in brain infections and neuro-inflammatory pathologies. However, access to such cells is limited, and studies systematically mapping the spectrum of their inflammatory states are scarce. Here, we generated microglia-like cells (MGLCs) from human induced pluripotent stem cells and characterized them as a robust, accessible model system for studying inflammatory activation. We validated lineage identity through transcriptome profiling, revealing selective upregulation of microglial signature genes and enrichment of microglia/macrophage-related gene sets. MGLCs displayed distinct morphologies and produced stimulus- and time-dependent cytokine secretion profiles upon exposure to diverse inflammatory stimuli, including pro-inflammatory cytokines (TNFα, interferon-γ) and agonists of the Toll-like receptors TLR2 (FSL-1), TLR3 (Poly(I:C)), TLR4 (lipopolysaccharide, LPS), and TLR7 (imiquimod). Transcriptome profiling and bioinformatics analysis revealed distinct activation signatures. Functional assays demonstrated stimulus-specific engagement of NFκB and JAK-STAT signaling pathways. The shared NFκB nuclear translocation response of TLR ligands and TNFα was reflected in overlapping transcriptome profiles: they shared modules (e.g., oxidative stress response and TNFα-related signaling) identified by weighted gene co-expression network analysis. Finally, the potential consequences of microglia activation for neighboring cells were studied on the example of microglia-astrocyte crosstalk. The capacity of MGLC supernatants to stimulate astrocytes was measured by quantifying astrocytic NFκB translocation. MGLCs stimulated with FSL-1, LPS, or Poly(I:C) indirectly activated astrocytes via a strictly TNFα-dependent mechanism, highlighting the role of soluble mediators in the signal propagation. Altogether, this platform enables a dissection of microglia activation states and multi-parametric characterization of subsequent neuroinflammation.
    Keywords:  (neuro-)inflammation; TLR; TNFα; astrocytes; cytokine release; transcriptome changes
    DOI:  https://doi.org/10.3390/cells14211687
  19. STAR Protoc. 2025 Nov 12. pii: S2666-1667(25)00605-7. [Epub ahead of print]6(4): 104199
      Autofluorescence in brain tissue poses a challenge in immunofluorescent staining by obscuring antibody-labeled proteins, mainly within microglia. Here, we describe a cost-effective protocol for removing autofluorescence in mouse brain sections by photobleaching. We outline steps to collect and section brain tissue, apply a photobleaching step using a light-emitting diode (LED), and perform immunofluorescent staining with primary and secondary antibodies. This protocol enables clearer visualization of microglial markers, particularly in mouse models of neurodegenerative diseases and aging.
    Keywords:  antibody; genetics; immunology; microscopy; model organisms; molecular/chemical probes; neuroscience
    DOI:  https://doi.org/10.1016/j.xpro.2025.104199
  20. Biotechnol Bioeng. 2025 Nov 12.
      Microglia are critical regulators of brain homeostasis and immune responses in the central nervous system (CNS). However, existing human-based models fail to reproduce the early and complex microglia-neural cell interactions. The differentiation of human induced pluripotent stem cells (hiPSCs) into specialized cell types offers promising avenues for understanding human development and disease modeling. Herein, a methodology for the differentiation of hiPSC-derived erythromyeloid progenitors (iEMPs) and their 3D co-culture with hiPSC-derived neurospheres were explored, utilizing the Ambr 250 Modular stirred-tank bioreactor (STB) system. The aim of this study was to build a complex co-culture model between iEMP and neurospheres in a scalable and controlled environment. Our results demonstrate that the STB effectively supports the co-culture process, with iEMP integration into the neurospheres, exhibiting cell density, aggregate morphology, and concentration similar to the neurosphere cultures. The co-culture environment induced the upregulation of transcription factors critical for microglial lineage commitment. iEMP-neurospheres displayed a unique secretory profile, releasing proteins involved in extracellular matrix remodeling and neuronal differentiation, essential for microenvironment remodeling. In conclusion, this study underscores the role of iEMPs in CNS development and presents a robust platform for preclinical research.
    Keywords:  hiPSCs; iEMP; neurospheres; stem cell bioengineering; stirred‐tank bioreactors
    DOI:  https://doi.org/10.1002/bit.70100
  21. Bio Protoc. 2025 Nov 05. 15(21): e5493
      Three-dimensional (3D) human brain tissue models derived from induced pluripotent stem cells (iPSCs) have transformed the study of neural development and disease in vitro. While cerebral organoids offer high structural complexity, their large size often leads to necrotic core formation, limiting reproducibility and challenging the integration of microglia. Here, we present a detailed, reproducible protocol for generating multi-cell type 3D neurospheres that incorporate neurons, astrocytes, and optionally microglia, all derived from the same iPSCs. While neurons and astrocytes differentiate spontaneously from neural precursor cells, generated by dual SMAD-inhibition (blocking BMP and TGF-b signaling), microglia are generated in parallel and can infiltrate the mature neurosphere tissue after plating neurospheres into 48-well plates. The system supports a range of downstream applications, including functional confocal live imaging of GCaMP6f after adeno-associated virus (AAV) transduction of neurospheres or immunofluorescence staining after fixation. Our approach has been successfully implemented across multiple laboratories, demonstrating its robustness and translational potential for studying neuron-glia interactions and modeling neurodegenerative processes. Key features • Reproducible human iPSC-derived 3D neurosphere multi-cell type tissue culture system. • Optional addition of microglia allows for studying neuron-microglia interaction in vitro in 3D. • Reliable spontaneous activity offers functional tissue culture readouts of neural firing. • System allows modeling of human brain diseases, such as Alzheimer's disease.
    Keywords:  3D neural tissue; In vitro disease modeling; Microglia; Neuro–glia interaction; iPSC-derived cells
    DOI:  https://doi.org/10.21769/BioProtoc.5493