Abstract

Astrocytes are the most abundant cell type in the brain, where they perform a wide array of functions, yet the nature of their cellular heterogeneity and how it oversees these diverse roles remains shrouded in mystery. Using an intersectional fluorescence-activated cell sorting–based strategy, we identified five distinct astrocyte subpopulations present across three brain regions that show extensive molecular diversity. Application of this molecular insight toward function revealed that these populations differentially support synaptogenesis between neurons. We identified correlative populations in mouse and human glioma and found that the emergence of specific subpopulations during tumor progression corresponded with the onset of seizures and tumor invasion. In sum, we have identified subpopulations of astrocytes in the adult brain and their correlates in glioma that are endowed with diverse cellular, molecular and functional properties. These populations selectively contribute to synaptogenesis and tumor pathophysiology, providing a blueprint for understanding diverse astrocyte contributions to neurological disease.

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Acknowledgements

We thank M. Brenner and M. Goodell for assistance with our cell-surface-marker antibody screen. This work was supported by grants from the Sontag Foundation (B.D.), the National Multiple Sclerosis Society (RG-1501-02756; B.D.), the Cancer Prevention Research Institute of Texas (RP510334 and RP160192 awarded to both B.D. and C.J.C.), the American Cancer Society (PF-15-220; K.Y.) and the US National Institutes of Health (NIH) (NS071153 and AG054111 (B.D.), NS089366 (B.D.), NS29709 (J.L.N.) and T32HL902332 (K.Y. and J.C.)). This project was also supported in part by the Genomic and RNA Profiling Core at Baylor College of Medicine with funding from the NIH–NCI grant (P30CA125123) and the expert assistance of L. White, the Cytometry and Cell Sorting Core at Baylor College of Medicine with funding from the NIH (P30 AI036211, P30 CA125123 and S10 RR024574), the expert assistance of J. Sederstrom and by IDDRC grant number 1U54 HD083092 from the Eunice Kennedy Shriver National Institute of Child Health and Human Development.

Author information

Author notes

    • Asante Hatcher
    •  & Teng-Wei Huang

    These authors contributed equally to this work.

    • Chad J Creighton
    •  & Benjamin Deneen

    These authors jointly directed this work.

Affiliations

  1. Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas, USA.

    • Chia-Ching John Lin
    • , Kwanha Yu
    • , Teng-Wei Huang
    • , Jeffrey Carlson
    • , Wenyi Zhu
    • , Nabil Ahmed
    •  & Benjamin Deneen
  2. Department of Neuroscience, Baylor College of Medicine, Houston, Texas, USA.

    • Asante Hatcher
    • , Hyun Kyoung Lee
    • , Benjamin R Arenkiel
    • , Jeffrey L Noebels
    •  & Benjamin Deneen
  3. Neurological Research Institute at Texas' Children's Hospital, Baylor College of Medicine, Houston, Texas, USA.

    • Hyun Kyoung Lee
    • , Akash J Patel
    • , Benjamin R Arenkiel
    •  & Benjamin Deneen
  4. Department of Pediatrics, Division of Neurology, Texas Children's Hospital, Houston, Texas, USA.

    • Hyun Kyoung Lee
  5. Program in Developmental Biology, Baylor College of Medicine, Houston, Texas, USA.

    • Jeffrey Carlson
    • , Benjamin R Arenkiel
    •  & Benjamin Deneen
  6. Department of Neurological Sciences, University of Vermont, Vermont, Vermont, USA.

    • Matthew C Weston
  7. Dan L. Duncan Cancer Center, Division of Biostatistics, Baylor College of Medicine, Houston, Texas, USA.

    • Fengju Chen
    • , Yiqun Zhang
    •  & Chad J Creighton
  8. Department of Pathology, Texas Children's Hospital, Baylor College of Medicine, Houston, Texas, USA.

    • Carrie A Mohila
  9. Department of Neurosurgery, Baylor College of Medicine, Houston, Texas, USA.

    • Akash J Patel
  10. Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas, USA.

    • Benjamin R Arenkiel
    •  & Jeffrey L Noebels
  11. Department of Neurology, Baylor College of Medicine, Houston, Texas, USA.

    • Jeffrey L Noebels
  12. Department of Medicine, Baylor College of Medicine, Houston, Texas, USA.

    • Chad J Creighton

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Contributions

C.-C.J.L., K.Y. and B.D. conceived the project, designed the experiments and wrote the manuscript; C.-C.J.L., K.Y., A.H., T.-W.H., H.K.L., J.C. and W.Z. performed the experiments; M.C.W., C.A.M., N.A., A.J. and B.R.A. provided essential reagents; F.C., Y.Z. and C.J.C. designed and executed the bioinformatics analysis; and J.L.N. and C.J.C. assisted in experimental design.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Chad J Creighton or Benjamin Deneen.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–6

  2. 2.

    Supplementary Methods Checklist

Excel files

  1. 1.

    Supplementary Table 1

    List of Cell Surface Antibodies; Statistical Analysis of Astrocyte Subpopulation Dynamics

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    Supplementary Table 2

    Astrocyte and Neuron Control Gene List

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    Supplementary Table 3

    RNA-Seq Statistical Analysis, Pop v Neg Gene Lists

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    Supplementary Table 4

    Pop v Neg Gene List GO Analysis

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    Supplementary Table 5

    RNA-Seq Statistical Analysis, Pop v Pop Gene Lists

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    Supplementary Table 6

    Pop v Pop Gene List GO Analysis

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    Supplementary Table 7

    GSEA derived Population C Synpase gene list and GO

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    Supplementary Table 8

    Glioma Population Specific Gene Lists

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    Supplementary Table 9

    Astrocyte Subpopulation Gene Expression in Glioma Populations; Epilepsy GSEA analysis gene list

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    Supplementary Table 10

    Subject Genders and Ages

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    Supplementary Table 11

    Antibody Information

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DOI

https://doi.org/10.1038/nn.4493

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