Biology subjects

Nobuta, H.

Publications and source records attributed to Nobuta, H..

3 recordsLinked to original sources

bioRxiv2026

Transcriptional Heterogeneity Reveals a Synaptic Gene Program in Developing and Adult Human Oligodendrocyte Precursor Cells

Human oligodendrocyte precursor cells (OPCs) arise early in gestation and expand broadly during cortical development, yet the extent of their heterogeneity remains poorly defined. Here, we isolated >2,300 highly pure OPCs from post-conceptional week (PCW) 17 human cortex using an optimized PDGFR-based immunopanning and performed single-cell RNA sequencing. Unsupervised clustering revealed four transcriptionally distinct embryonic OPC subsets, including a previously unrecognized population that expressed genes linked to synaptic development, synaptic signaling, and neuromodulation. This subset - designated embryonic synaptic OPCs (eSyn-OPCs) - comprised approximately 28.5% of all embryonic OPCs in the cortex and was characterized by robust expression of synapse-associated secreted factors (THBS2, WNT5A, WNT7A, PLAT, ACHE) and multiple neurotransmitter receptor subunits. Histological analyses across PCW 12-22 demonstrated that eSyn-OPCs first appear around PCW 15 and are enriched in proliferative germinal zones. Spatial transcriptomics confirmed their localization near neural stem and progenitor cells, suggesting proximal neuron-OPC communication during early cortical assembly. Purified eSyn-OPCs differentiated into mature oligodendrocytes in vitro, confirming their oligodendrocyte lineage identity. Reanalysis of adult human single-nucleus RNA-seq datasets uncovered a transcriptionally analogous OPC subset (adult synaptic OPCs, aSyn-OPCs), though with reduced representation of structural synaptic genes and neurotransmitter receptor diversity compared to eSyn-OPCs. Together, these results identify a synaptically specialized OPC population in both developing and adult human cortex and reveal that eSyn-OPCs possess unexpectedly rich synaptic signaling machinery. These findings suggest that human OPCs may participate directly in neuron-glia communication during early cortical development and raise the possibility of developmental stage-specific roles for eSyn-OPCs in shaping neural circuit formation.

neuroscience↗

bioRxiv2025

Controlled Delivery of a Neurotrophic Factor in the Adult Mouse Brain Using Engineered Microglia

Microglia, the resident immune cells of the central nervous system, have been proposed as vehicles for delivering therapeutic biologics. These cells can be genetically engineered in vitro and transplanted into host animals following ablation of endogenous microglia, enabling repopulation of the brain parenchyma. However, current replacement strategies often rely on radiation or transgenic models, limiting their clinical relevance. CSF1R inhibitors offer a more translational approach to microglia ablation, though surviving host cells can compete with transplanted microglia during repopulation. In this study, we successfully ablated endogenous microglia using a CSF1R inhibitor in adult mice and developed a method to transplant engineered microglia expressing Brain-Derived Neurotrophic Factor (BDNF) in a doxycycline-inducible manner. To enhance engraftment, transplanted cells also expressed a constitutively active CSF1R mutant (caCSF1R). BDNF-expressing transplanted microglia spread through large areas of host mice brains, displayed similar morphology and transcriptional profile to repopulating host microglia, and responded to pro-inflammatory stimuli. Treatment with doxycycline resulted in increased BDNF expression and TrkB phosphorylation in the host brain. Expression of caCSF1R provided transplanted cells with a competitive advantage over endogenous repopulating cells, resulting in the accelerated spread of the transplants. Our results demonstrate the functional integration and therapeutic potential of microglia as vehicles for delivering neurotrophic factors to the brain in a controllable manner. Furthermore, we show that caCSF1R expression is able to enhance the spread of transplanted microglia. SIGNIFICANCEThis study demonstrates the potential of engineered microglia to deliver the protein Brain-Derived Neurotrophic Factor to the brain parenchyma, under the control of orally-administered doxycycline. The technique can be generalized to a wide array of proteins, offering a novel paradigm for neurological therapy.

neuroscience↗

bioRxiv2025

The mGluR5 Agonist, CHPG, Enhances Numbers of Differentiated Human Oligodendrocytes

Previous studies on adult mice indicate that the mGluR5 agonist 2-chloro-5-hydroxyphenyl glycine (CHPG), reduces cuprizone-elicited losses in myelin. This effect is partly mediated by CHPG binding to mGluR5 receptors on reactive astrocytes, triggering the release of brain derived neurotrophic factor (BDNF), which results in an increase in myelin, and alleviates behavioral deficits. However, it remains unclear whether CHPG has similar beneficial effects on human cells. To address this issue, we examined effects of CHPG on human cells using both human induced pluripotent stem cell (hiPSC)-derived oligodendrocytes and primary human fetal brain cells. Treatment of hiPSCs (30M, 5 days) or primary cells (30 M, 3 days) with CHPG increases the percent of MBP+O4+ mature oligodendrocytes relative to total O4+cells, without affecting survival. When effects of CHPG were evaluated on proliferating OPCs, effects on proliferation are observed. In contrast, when CHPG was evaluated in young oligodendrocytes, effects on proliferation were gone, suggesting that in this population CHPG is influencing differentiation. Interestingly, in contrast to observations in mice, mGluR5 expression in humans is localized on PDGFR+ oligodendrocyte precursor cells (OPCs) and O4+ immature oligodendrocytes, but not astrocytes. Moreover, using purified human OPC cultures, we show a direct effect of CHPG in enhanced differentiation. To identify potential cellular targets of CHPG in the adult human brain, we analyzed postmortem tissue from individuals with multiple sclerosis (MS) and healthy controls. In contrast to the hiPSCs or fetal cells, demyelinated white matter from MS patients showed elevated mGluR5 mRNA expression in astrocytes. Taken together, our findings suggest that CHPG enhances the differentiation of human OPCs during development through a mechanism distinct from that observed in adult cuprizone-treated mice. Moreover, astrocytes in MS pathology upregulate mGluR5, suggesting they may become responsive to CHPG under disease conditions.

neuroscience↗