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Astrocytic trans-differentiation completes a multicellular paracrine feedback loop required for medulloblastoma tumor growth

ABSTRACT: Astrocytic trans-differentiation completes a multicellular paracrine feedback loop required for medulloblastoma tumor growth

PROVIDER: PRJNA437986 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6827140_1.fastq.gz	Fastqsanger.gz
	SRR6827140_2.fastq.gz	Fastqsanger.gz
	SRR6827150_1.fastq.gz	Fastqsanger.gz
	SRR6827150_2.fastq.gz	Fastqsanger.gz
	SRR6827160_1.fastq.gz	Fastqsanger.gz

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Astrocytic trans-differentiation completes a multicellular paracrine feedback loop required for medulloblastoma tumor growth

Project description:Astrocytic trans-differentiation completes a multicellular paracrine feedback loop required for medulloblastoma tumor growth

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Astrocytic trans-Differentiation Completes a Multicellular Paracrine Feedback Loop Required for Medulloblastoma Tumor Growth.

Project description:The tumor microenvironment (TME) is critical for tumor progression. However, the establishment and function of the TME remain obscure because of its complex cellular composition. Using a mouse genetic system called mosaic analysis with double markers (MADMs), we delineated TME evolution at single-cell resolution in sonic hedgehog (SHH)-activated medulloblastomas that originate from unipotent granule neuron progenitors in the brain. First, we found that astrocytes within the TME (TuAstrocytes) were trans-differentiated from tumor granule neuron precursors (GNPs), which normally never differentiate into astrocytes. Second, we identified that TME-derived IGF1 promotes tumor progression. Third, we uncovered that insulin-like growth factor 1 (IGF1) is produced by tumor-associated microglia in response to interleukin-4 (IL-4) stimulation. Finally, we found that IL-4 is secreted by TuAstrocytes. Collectively, our studies reveal an evolutionary process that produces a multi-lateral network within the TME of medulloblastoma: a fraction of tumor cells trans-differentiate into TuAstrocytes, which, in turn, produce IL-4 that stimulates microglia to produce IGF1 to promote tumor progression.

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Astrocytic trans-differentiation completes a multicellular paracrine feedback loop required for medulloblastoma tumor growth

Project description:We used RNA-sequencing technology for high-throughput profiling of tumor-derived astrocytes. Laser capture microdissection of individual cells was performed on an ArcturusXT LCM system (Applied Biosystems) with an infrared laser using CapSure HS LCM caps after optimization of laser power and duration. For each sample, 250 cells were microdissected per LCM cap and split into five 50-cell technical replicates (as a control, one replicate did not undergo reverse transcription) after elution from the cap. Reamplified cDNA (~500 bp 3’ ends) was purified away from primer concatemers by two rounds of purification with 0.7x volume of AMPure XP beads (Beckman) according to the manufacturer’s recommendations, and purified cDNA was quantified by Qubit assay (Life Technologies). For each sample, 1 ng was tagmented with the Nextera XT kit (Illumina) and sequenced as 75 bp paired-end reads on a NextSeq instrument using v2 reagents (Illumina). 14–22 million reads per sample were filtered for signal to noise (chastity filtered), assessed for overall quality with FastQC (http://www.bioinformatics.babraham.ac.uk/projects/fastqc/), and then mapped with STAR (Dobin et al., 2013) against the mouse genome build mm10.

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Astrocytic trans-differentiation completes a multicellular paracrine feedback loop required for medulloblastoma tumor growth

Project description:We report the application of RNA-sequencing technology for high-throughput profiling of tumor-associated microglia/macrophage (TAMs). First, we checked the purity of sample by qRT-PCR. We confirmed the high purity of TAMs with rare other cell type contamination. By profiling samples of TAMs purified from five individual medulloblastoma of mouse model, we find that TAMs maintain microglia gene signature even there are decrease of microglia genes and increase of macrophage genes. This study provides the evidence for the application of comprehensive gene profiling towards characterization of cellular origin of cell populations in tumor microenvironment.

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Expression profiling of lower-grade diffuse astrocytic glioma

Project description:Diffuse gliomas represent the most prevalent class of primary brain tumor. Despite significant recent advances in the understanding of glioblastoma (WHO IV), its most malignant subtype, lower-grade (WHO II and III) glioma variants remain comparatively understudied, especially in light of their notably variable clinical behavior. To examine the foundations of this heterogeneity, we performed multidimensional molecular profiling, including global transcriptional analysis, on 101 lower-grade diffuse astrocytic gliomas collected at our own institution, and validated our findings using publically available gene expression and copy number data from large independent patient cohorts. We found that IDH mutational status delineated molecularly and clinically distinct glioma subsets, with IDH mutant (IDH mt) tumors exhibiting TP53 mutations, PDGFRA overexpression, and prolonged survival, and IDH wild-type (IDH wt) tumors exhibiting EGFR amplification, PTEN loss, and unfavorable disease outcome. Furthermore, global expression profiling revealed three robust molecular subclasses within lower-grade diffuse astrocytic gliomas, two of which were predominantly IDH mt and one almost entirely IDH wt. IDH mt subclasses were distinguished from each other on the basis of TP53 mutations, DNA copy number abnormalities, and links to distinct stages of neurogenesis in the subventricular zone (SVZ). This latter finding implicates discrete pools of neuroglial progenitors as cells of origin for the different subclasses of IDH mt tumors. In summary, we have elucidated molecularly distinct subclasses of lower-grade diffuse astrocytic glioma that dictate clinical behavior and demonstrate fundamental associations with both IDH mutational status and neuroglial developmental stage. 80 tumor samples, one normal tissue sample (brain)

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