Small-molecule screen reveals pathways that regulate C4 secretion in stem cell-derived astrocytes

Stem Cell Reports, vol. 18, pp. 237–253

Abstract

In the brain, the complement system plays a crucial role in the immune response and in synaptic elimination during normal development and disease. Here, we sought to identify pathways that modulate the production of complement component 4 (C4), recently associated with an increased risk of schizophrenia. To design a disease-relevant assay, we first developed a rapid and robust 3D protocol capable of producing large numbers of astrocytes from pluripotent cells. Transcriptional profiling of these astrocytes confirmed the homogeneity of this population of dorsal fetal-like astrocytes. Using a novel ELISA-based small-molecule screen, we identified epigenetic regulators, as well as inhibitors of intracellular signaling pathways, able to modulate C4 secretion from astrocytes. We then built a connectivity map to predict and validate additional key regulatory pathways, including one involving c-Jun-kinase. This work provides a foundation for developing therapies for CNS diseases involving the complement cascade.

Authors 22

1. Francesca A. Rapino corresponding

   Broad Institute · Harvard University · Harvard Stem Cell Institute · Stanley Center for Psychiatric Research

   Affiliation as printed

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: francesca_rapino@harvard.edu

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

   Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

   Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

2. Ted E. Natoli

   Broad Institute

   Affiliation as printed

   Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

3. Francesco Limone Aachen

   Broad Institute · Harvard University · Leiden University Medical Center · Harvard Stem Cell Institute · Stanley Center for Psychiatric Research

   Affiliation as printed

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Leiden University Medical Center, LUMC, 2333 ZA Leiden, the Netherlands

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

   Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

   Leiden University Medical Center, LUMC, 2333 ZA Leiden, the Netherlands

   Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

4. Jack Blank

   Harvard University · Harvard Stem Cell Institute

   Affiliation as printed

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

   Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

5. Matthew Tegtmeyer

   Broad Institute · Harvard University · Stanley Center for Psychiatric Research · Massachusetts Institute of Technology

   Affiliation as printed

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

   Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

6. William C. Chen

   Harvard University · Harvard Stem Cell Institute

   Affiliation as printed

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

   Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

7. Erika M. Norabuena

   Harvard University · Harvard Stem Cell Institute

   Affiliation as printed

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

   Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

8. Juhi G. Narula

   Harvard University · Harvard Stem Cell Institute

   Affiliation as printed

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

   Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

9. Dane Z. Hazelbaker

   Broad Institute · Harvard University · Stanley Center for Psychiatric Research · Massachusetts Institute of Technology

   Affiliation as printed

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

   Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

   Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

10. Gabriella Angelini

    Broad Institute · Harvard University · Stanley Center for Psychiatric Research · Massachusetts Institute of Technology

    Affiliation as printed

    Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

    Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

    Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

11. Lindy E. Barrett

    Broad Institute · Harvard University · Stanley Center for Psychiatric Research · Massachusetts Institute of Technology

    Affiliation as printed

    Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

    Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

    Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

12. Alison O’Neil

    Harvard University · Harvard Stem Cell Institute

    Affiliation as printed

    Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

    Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

13. Ursula Konstantin Beattie

    Harvard University · Harvard Stem Cell Institute

    Affiliation as printed

    Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

    Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

    Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

14. Jessica M. Thanos

    Harvard University · Massachusetts General Hospital

    Affiliation as printed

    Center for Quantitative Health, Center for Genomic Medicine and Department of Psychiatry, Massachusetts General Hospital, Boston, MA, USA; Department of Psychiatry, Harvard Medical School, Boston, MA, USA

    Center for Quantitative Health, Center for Genomic Medicine and Department of Psychiatry, Massachusetts General Hospital, Boston, MA, USA

    Department of Psychiatry, Harvard Medical School, Boston, MA, USA

15. Heather de Rivera

    Broad Institute · Harvard University · Stanley Center for Psychiatric Research

    Affiliation as printed

    Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Department of Genetics, Harvard Medical School, Boston, MA, USA

    Department of Genetics, Harvard Medical School, Boston, MA, USA

    Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

16. Steven D. Sheridan

    Harvard University · Massachusetts General Hospital

    Affiliation as printed

    Center for Quantitative Health, Center for Genomic Medicine and Department of Psychiatry, Massachusetts General Hospital, Boston, MA, USA; Department of Psychiatry, Harvard Medical School, Boston, MA, USA

    Center for Quantitative Health, Center for Genomic Medicine and Department of Psychiatry, Massachusetts General Hospital, Boston, MA, USA

    Department of Psychiatry, Harvard Medical School, Boston, MA, USA

17. Roy H. Perlis

    Harvard University · Massachusetts General Hospital

    Affiliation as printed

    Center for Quantitative Health, Center for Genomic Medicine and Department of Psychiatry, Massachusetts General Hospital, Boston, MA, USA; Department of Psychiatry, Harvard Medical School, Boston, MA, USA

    Center for Quantitative Health, Center for Genomic Medicine and Department of Psychiatry, Massachusetts General Hospital, Boston, MA, USA

    Department of Psychiatry, Harvard Medical School, Boston, MA, USA

18. Steven A. McCarroll

    Broad Institute · Harvard University · Stanley Center for Psychiatric Research

    Affiliation as printed

    Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Department of Genetics, Harvard Medical School, Boston, MA, USA

    Department of Genetics, Harvard Medical School, Boston, MA, USA

    Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

19. Beth A. Stevens

    Broad Institute · Boston Children's Hospital · Harvard University · Stanley Center for Psychiatric Research

    Affiliation as printed

    Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Department of Neurology, F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA

    Department of Neurology, F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA

    Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

20. Aravind N. Subramanian

    Broad Institute

    Affiliation as printed

    Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

21. Ralda Nehme

    Broad Institute · Harvard University · Stanley Center for Psychiatric Research · Massachusetts Institute of Technology

    Affiliation as printed

    Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

    Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

    Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA

22. Lee L. Rubin corresponding

    Broad Institute · Harvard University · Harvard Stem Cell Institute · Stanley Center for Psychiatric Research

    Affiliation as printed

    Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA; Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: lee_rubin@harvard.edu

    Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA

    Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA

    Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA