Project description:This study explores the circuit integration of human glioblastoma organoids (GBOs) in vivo in the adult mouse brain. We performed single cell RNA sequencing (scRNA-seq) to understand the cell state diversity of malignant tumor cells in GBOs at baseline and after stimulation by 1 mM acetylcholine (ACh) for 1 hour. Sliced neocortical organoids (SNOs) were also sequenced to study gene expression properties of neural stem cells (NSCs).
Project description:This study explores the circuit integration of human glioblastoma organoids (GBOs) in vivo in the adult mouse brain. Here, we performed RNA sequencing analysis of GBOs at baseline conditions and various treatment timepoints of 1 mM acetylcholine (ACh).
Project description:Neuronal activity-driven mechanisms impact glioblastoma cell proliferation and invasion (1–7), and glioblastoma remodels neuronal circuits (8,9). Distinct intratumoral regions maintain functional connectivity via a subpopulation of malignant cells that mediate tumor-intrinsic neuronal connectivity and synaptogenesis through their transcriptional programs (8). However, the effects of tumor-intrinsic neuronal activity on other cells, such as immune cells, remain unknown. Here we show that regions within glioblastomas displaying elevated connectivity are characterized by regional immunosuppression. This is accompanied by different cell compositions and inflammatory status of tumor-associated macrophages (TAMs) in the tumor microenvironment. In preclinical models, genetic knockout of Thrombospondin-1 (TSP1/Thbs1), a synaptogenic factor critical for glioma-induced neuronal circuit remodeling, in glioblastoma cells suppressed synaptogenesis and glutamatergic neuronal hyperexcitability. Moreover, this restored antigen-presentation and pro-inflammatory responses, promoted the infiltration of pro-inflammatory TAMs and CD8+ T-cells, and mitigated the immunosuppressive effect of TAMs on T-cell proliferation. Furthermore, pharmacological inhibition of glutamatergic excitatory neuronal signaling redirected TAMs toward a less immunosuppressive phenotype, resulting in prolonged mouse survival. Lastly, pharmacological inhibition of glutamatergic signaling showed potential to enhance the efficacy of immune cell-based therapy. Altogether, our results demonstrate previously unrecognized immunosuppression mechanisms resulting from glioma-neuronal circuit remodeling and suggest that targeting glioma-neuron-immune crosstalk could provide new avenues for immunotherapy.
Project description:Neuronal activity-driven mechanisms impact glioblastoma cell proliferation and invasion (1–7), and glioblastoma remodels neuronal circuits (8,9). Distinct intratumoral regions maintain functional connectivity via a subpopulation of malignant cells that mediate tumor-intrinsic neuronal connectivity and synaptogenesis through their transcriptional programs (8). However, the effects of tumor-intrinsic neuronal activity on other cells, such as immune cells, remain unknown. Here we show that regions within glioblastomas displaying elevated connectivity are characterized by regional immunosuppression. This is accompanied by different cell compositions and inflammatory status of tumor-associated macrophages (TAMs) in the tumor microenvironment. In preclinical models, genetic knockout of Thrombospondin-1 (TSP1/Thbs1), a synaptogenic factor critical for glioma-induced neuronal circuit remodeling, in glioblastoma cells suppressed synaptogenesis and glutamatergic neuronal hyperexcitability. Moreover, this restored antigen-presentation and pro-inflammatory responses, promoted the infiltration of pro-inflammatory TAMs and CD8+ T-cells, and mitigated the immunosuppressive effect of TAMs on T-cell proliferation. Furthermore, pharmacological inhibition of glutamatergic excitatory neuronal signaling redirected TAMs toward a less immunosuppressive phenotype, resulting in prolonged mouse survival. Lastly, pharmacological inhibition of glutamatergic signaling showed potential to enhance the efficacy of immune cell-based therapy. Altogether, our results demonstrate previously unrecognized immunosuppression mechanisms resulting from glioma-neuronal circuit remodeling and suggest that targeting glioma-neuron-immune crosstalk could provide new avenues for immunotherapy.
Project description:Gene expression profiling of distinct members of a neuronal circuit has the potential to identify candidate molecules and mechanisms that underlie the formation, organization and function of the circuit. To this end, we report here the application of a novel method to characterize RNAs from small numbers of specific Drosophila brain neurons, which belong to the circadian circuit. We identified three different sets of mRNAs enriched in different subclasses of clock neurons: one is enriched in all clock neurons, a second is enriched in PDF-positive clock neurons and a third is enriched in PDF-negative clock neurons. Moreover, we characterized 2 novel genes, Fer2 and dnocturnin, one from each subgroup, which highlight subgroup-specific features and play important roles in circadian rhythms. The methodology is a powerful tool not only to dissect the cellular and molecular basis of circadian rhythms but also to molecularly characterize other Drosophila neuronal circuits. Experiment Overall Design: Circadican related neuronal celltypes (Tim, Pdf) or general neurons (Elav) were labeled by GFP or YFP using specific Gal4 drivers. Expression of those celltypes were profiled after manual sorting of those GFP or YFP positive cells. 3 biological replicates were collected (except adult small pdf cells).
Project description:The molecular etiology of numerous risk genes for autism spectrum disorder (ASD), including CDH11, remains elusive. We investigated the role of CDH11 in the development of ASD-related behaviors using gene-deficient mice. CDH11 is enriched at synapses in glutamatergic neurons of the anterior cingulate cortex (ACC), which project to the striatum, nucleus accumbens, and amygdala. Ablation of Cdh11 in these neurons during development increases self-grooming and reduces sociability, with decreased neuronal activity in the ACC. Chemogenetic inhibition of ACC glutamatergic neurons recapitulates the over-grooming phenotype, while activation of these neurons mitigates self-grooming in Cdh11-deficient mice. Proteomics of ACC synaptosomes and CDH11 interactomes suggest the involvement of CDH11 in synaptic vesicle trafficking, supported by reduced presynaptic vesicle density at excitatory synapses in Cdh11-deficient mice. These findings highlight the critical role of CDH11 in ASD-related brain circuit development and offer insights into the molecular mechanisms and potential therapeutic targets for ASD.
Project description:The molecular etiology of numerous risk genes for autism spectrum disorder (ASD), including CDH11, remains elusive. We investigated the role of CDH11 in the development of ASD-related behaviors using gene-deficient mice. CDH11 is enriched at synapses in glutamatergic neurons of the anterior cingulate cortex (ACC), which project to the striatum, nucleus accumbens, and amygdala. Ablation of Cdh11 in these neurons during development increases self-grooming and reduces sociability, with decreased neuronal activity in the ACC. Chemogenetic inhibition of ACC glutamatergic neurons recapitulates the over-grooming phenotype, while activation of these neurons mitigates self-grooming in Cdh11-deficient mice. Proteomics of ACC synaptosomes and CDH11 interactomes suggest the involvement of CDH11 in synaptic vesicle trafficking, supported by reduced presynaptic vesicle density at excitatory synapses in Cdh11-deficient mice. These findings highlight the critical role of CDH11 in ASD-related brain circuit development and offer insights into the molecular mechanisms and potential therapeutic targets for ASD
Project description:The molecular etiology of numerous risk genes for autism spectrum disorder (ASD), including CDH11, remains elusive. We investigated the role of CDH11 in the development of ASD-related behaviors using gene-deficient mice. CDH11 is enriched at synapses in glutamatergic neurons of the anterior cingulate cortex (ACC), which project to the striatum, nucleus accumbens, and amygdala. Ablation of Cdh11 in these neurons during development increases self-grooming and reduces sociability, with decreased neuronal activity in the ACC. Chemogenetic inhibition of ACC glutamatergic neurons recapitulates the over-grooming phenotype, while activation of these neurons mitigates self-grooming in Cdh11-deficient mice. Proteomics of ACC synaptosomes and CDH11 interactomes suggest the involvement of CDH11 in synaptic vesicle trafficking, supported by reduced presynaptic vesicle density at excitatory synapses in Cdh11-deficient mice. These findings highlight the critical role of CDH11 in ASD-related brain circuit development and offer insights into the molecular mechanisms and potential therapeutic targets for ASD.