scholarly journals TAMI-53. THE ONCOMETABOLITE D-2HG INDUCE INFLAMMATORY ASTROGLIOSIS CAUSING NEUROTOXICITY AND SEIZURES IN IDH MUTATED GLIOMA

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii224-ii225
Author(s):  
Simon Behringer ◽  
Vidhya Ravi ◽  
Kevin Joseph ◽  
Juergen Beck ◽  
Oliver Schnell ◽  
...  
Keyword(s):  
Rna Sequencing ◽  
Metabolic Alteration ◽  
Neocortical Slice ◽  
Reactive Trajectories ◽  
Tumor Environment ◽  
Single Nucleus ◽  
Inflammatory Pattern ◽  
Transcriptional Adaptation ◽  
Transcriptional Phenotype

Abstract The role of tumor-associated astrocytes in the microenvironment of glioma has long been underestimated but is moving into the focus of current research. We explored the role of reactive astrocytes in IDH-mutated glioma using RNA-sequencing of purified astrocytes and microglia and single-nucleus RNA-sequencing of infiltrating tumor regions. Mapping of the transcriptional phenotype of astrocytes along developmental and reactive trajectories revealed an inflammatory transformation of IDH-mutated associated astrocytes. The major proportion of astrocytes is marked by complement-activation similar to findings in neuroinflammatory diseases. A human neocortical slices model with injected IDH-mutated patient-derived cells or D-2HG treatment (+/- microglia depletion) was used to map shared and unique transcriptional adaptation in astrocytes promoted by either tumor cells or metabolic alteration. High-dimensional electrophysiological profiling was used to investigate alterations in neural response to tumor-induced microenvironmental transformation. We showed that 2HG alone promote the inflammatory pattern of astrocytes, which causes neurotoxicity and seizures in our neocortical slice model. Depletion of microglia rescued the neurotoxicity suggesting that microglia predominantly drive inflammatory astrogliosis as a response to metabolic alteration the tumor environment. We showed that neurotoxic astrogliosis induced by the oncometabolite D-2HG via distinct microglia activation promote the evolution of frequently observed seizures in IDH-mutated glioma patients.

scholarly journals P11.58 Astrocytic Environment Aid the Evolution of Immunosuppressive Environment in Glioblastoma

2019 ◽  
Vol 21 (Supplement_3) ◽  
pp. iii57-iii57
Author(s):  
D H Heiland ◽  
V M Ravi ◽  
S P Behringer ◽  
O Schnell
Keyword(s):  
Rna Sequencing ◽  
De Novo ◽  
Inflammatory Diseases ◽  
Reactive Astrocytes ◽  
Loss Of Function ◽  
Cellular Environment ◽  
Pathway Activation ◽  
Tumor Environment ◽  
Anti Inflammatory ◽  
Transcriptional Phenotype

Abstract BACKGROUND Glioblastomas are referred to as immunologically “cold” tumors since their cellular environment promotes an anti-inflammatory environment leading to an unresolved barrier to immunotherapy. Although the role of reactive astrocytes in other inflammatory diseases has been investigated in several studies, the immunoregulatory functions of astrocytes in the tumor environment remains poorly understood. MATERIAL AND METHODS We purified reactive astrocytes from de-novo glioblastoma and non-infiltrated cortex specimens by immunoprecipitation and analyzed the transcriptional phenotype by RNA sequencing. In order to investigate the origin of astrocytic transformation, we used a microglia loss-of-function model in human organotypic slices. Microglia was depleted by clodronat stimulation. We injected tumor cells and analyzed gene expression of the astrocytes after 7d incubation by RNA sequencing. Environmental cytokines were analyzed by multi-ELISA. Immunostainings of slices were processed by confocal microscopy and 3D reconstruction. RESULTS Here we address the immunological impact of tumor-associated astrocytes, we were able to identify a novel reactive subtype marked by JAK/STAT pathway activation and CD274 expression. Our results show a distinct astrocytic transcriptional phenotype that mutually arises from both microglia and astrocytes of the tumor environment. This interaction leads to a large release of anti-inflammatory cytokines such as TGFß and IL10. The reactive subtype switch of astrocytes was recovered by inhibition of the JAK pathway, which caused an increase of pro-inflammatory environment. CONCLUSION Our results can form the basis for a novel therapeutic approach by directly targeting tumor-associated astrocytes in order to transformimmunologically “cold” into “warm” tumors.

Gut Microbiota and Antipsychotics Induced Metabolic Alteration

2019 ◽  
pp. 131-143
Author(s):  
Dong-Yu Kan ◽  
Su-Juan Li ◽  
Chen-Chen Liu ◽  
Ren-Rong Wu
Keyword(s):  
Gut Microbiota ◽  
Body Weight Gain ◽  
Neuropsychiatric Disorders ◽  
Elevated Risk ◽  
Metabolic Disturbance ◽  
Metabolic Dysfunction ◽  
Sequencing Technology ◽  
Severe Mental Disorder ◽  
Metabolic Alteration

Schizophrenia is a chronic and severe mental disorder with antipsychotics as primary medications, but the antipsychotic-induced metabolic side effects may contribute to the elevated risk of overall morbidity and mortality in patients with psych-iatric diseases. With the development in sequencing technology and bioinformatics, dysbiosis has been shown to contribute to body weight gain and metabolic dysfunction. However, the role of gut microbiota in the antipsychotic-induced metabolic alteration remains unknown. In this paper, we reviewed the recent studies of the gut microbiota with psychiatric disorders and antipsychotic-induced metabolic dysfunction. Patients with neuropsychiatric disorders may have a different composi-tion of gut microbiota compared with healthy controls. In addition, it seems that the use of antipsychotics is concurrently associated with both altered composition of gut microbiota and metabolic disturbance. Further study is needed to address the role of gut microbiota in the development of neuropsychiatric disorders and antipsychotic-induced metabolic disturbance, to develop novel therapeutics for both neuropsychiatric disorders and metabolic dysfunction.

scholarly journals The molecular taxonomy of primate amygdala via single-nucleus RNA-sequencing analysis

2021 ◽  
Author(s):  
Lei Zhang ◽  
Yanyong Cheng ◽  
Shihao Wu ◽  
Yufeng Lu ◽  
Zhenyu Xue ◽  
...  
Keyword(s):  
Rna Sequencing ◽  
Molecular Taxonomy ◽  
Sequencing Analysis ◽  
Single Nucleus

Single-nucleus RNA-sequencing of human choriodecidua before and after the onset of labor

Placenta ◽  
2021 ◽  
Vol 112 ◽  
pp. e67
Author(s):  
Léa Chicoisne ◽  
Muriel Andrieu ◽  
Céline Bertholle ◽  
Vaarany Karunanithy ◽  
Brigitte Izac ◽  
...  
Keyword(s):  
Rna Sequencing ◽  
Before And After ◽  
Single Nucleus

TAMI-72. DIVERSITY OF CELLULAR COMMUNICATION IN GLIOBLASTOMA

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi213-vi213
Author(s):  
Kevin Joseph ◽  
Lea Vollmer ◽  
Vidyha Ravi ◽  
Jürgen Beck ◽  
Ulrich Hofmann ◽  
...  
Keyword(s):  
Rna Sequencing ◽  
Cellular Signaling ◽  
Significant Inhibition ◽  
Intercellular Signaling ◽  
Scale Free ◽  
Transcriptional Dynamics ◽  
Transcriptional Adaptation ◽  
Electrophysiological Characterization ◽  
Electrical Signaling

Abstract OBJECTIVE Owing to recent advances in understanding of the active functional states exhibited within glioblastoma (GBM), intra-tumoral cellular signaling has moved into focus of neuro-oncology. In this study, we aim to explore the diversity of transcellular signaling and investigate correlations between transcriptional dynamics and functional signaling. METHODS Electrophysiological characterization of GBM was carried out using planar microelectrodes and Ca2+ imaging, in both 2D cell culture as well as in our novel human cortical GBM model. Exposure to physiologically relevant conditions present within the tumor was carried out to identify specific signaling cells of interest and capture the signaling diversity in response to environmental conditions. Transcriptional dynamics and plasticity were examined by means of scRNA-sequencing with CRISPR based perturbation, spatial transcriptomics and deep long-read RNA-sequencing. RESULTS Electrophysiological profiles of multiple primary GBM cell lines revealed characteristics of scale-free networks (R2=0.875), confirmed in both 2D culture as well as a human neocortical GBM model. When GBM was cultured in a “in-vivo” like environment, basal activity was significantly higher (50%, p=0.01). Cellular signaling was directly correlated to changes in the environment, like hypoxia or glutamatergic activation, and total inhibition of electrical signaling required the usage of synaptic inhibitors. Using single-cell RNA sequencing and proteomics, several synaptogenesis related genes were identified to play a crucial role in the lineage states present in GBM. CRISPR based perturbation of these genes resulted in alterations in cellular morphology and decreased cellular connectivity (p< 0.01), with loss of scale free features (R2=0.35), and transcriptomic loss of developmental lineages (FDR< 0.01), leading to significant inhibition of GBM stress response. CONCLUSION Our findings highlight the role of electrical signaling in glioblastoma. Cellular stressors induce intercellular signaling, leading to transcriptional adaptation suggesting that there exists a highly complex and powerful mechanism for dynamic transcriptional state adaptation.

scholarly journals Single nucleus and in situ RNA sequencing reveals cell topographies in the human pancreas

2020 ◽  
Author(s):  
Luca Tosti ◽  
Yan Hang ◽  
Olivia Debnath ◽  
Sebastian Tiesmeyer ◽  
Timo Trefzer ◽  
...  
Keyword(s):  
Rna Sequencing ◽  
Human Pancreas ◽  
Single Nucleus

scholarly journals Single Cell, Single Nucleus and Spatial RNA Sequencing of the Human Liver Identifies Hepatic Stellate Cell and Cholangiocyte Heterogeneity

2021 ◽  
Author(s):  
Tallulah S Andrews ◽  
Jawairia Atif ◽  
Jeff C Liu ◽  
Catia T Perciani ◽  
Xue-Zhong Ma ◽  
...  
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
Human Liver ◽  
Stellate Cell ◽  
Parenchymal Cell ◽  
Cell Types ◽  
Cell Populations ◽  
Healthy Human ◽  
Single Nucleus

The critical functions of the human liver are coordinated through the interactions of hepatic parenchymal and non-parenchymal cells. Recent advances in single cell transcriptional approaches have enabled an examination of the human liver with unprecedented resolution. However, dissociation related cell perturbation can limit the ability to fully capture the human liver's parenchymal cell fraction, which limits the ability to comprehensively profile this organ. Here, we report the transcriptional landscape of 73,295 cells from the human liver using matched single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA sequencing (snRNA-seq). The addition of snRNA-seq enabled the characterization of interzonal hepatocytes at single-cell resolution, revealed the presence of rare subtypes of hepatic stellate cells previously only seen in disease, and detection of cholangiocyte progenitors that had only been observed during in vitro differentiation experiments. However, T and B lymphocytes and NK cells were only distinguishable using scRNA-seq, highlighting the importance of applying both technologies to obtain a complete map of tissue-resident cell-types. We validated the distinct spatial distribution of the hepatocyte, cholangiocyte and stellate cell populations by an independent spatial transcriptomics dataset and immunohistochemistry. Our study provides a systematic comparison of the transcriptomes captured by scRNA-seq and snRNA-seq and delivers a high-resolution map of the parenchymal cell populations in the healthy human liver.

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