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Chemical genetics reveals a complex functional ground state of neural stem cells

Abstract

The identification of self-renewing and multipotent neural stem cells (NSCs) in the mammalian brain holds promise for the treatment of neurological diseases and has yielded new insight into brain cancer1,2,3. However, the complete repertoire of signaling pathways that governs the proliferation and self-renewal of NSCs, which we refer to as the 'ground state', remains largely uncharacterized. Although the candidate gene approach has uncovered vital pathways in NSC biology4,5,6,7,8, so far only a few highly studied pathways have been investigated. Based on the intimate relationship between NSC self-renewal and neurosphere proliferation8, we undertook a chemical genetic screen for inhibitors of neurosphere proliferation in order to probe the operational circuitry of the NSC. The screen recovered small molecules known to affect neurotransmission pathways previously thought to operate primarily in the mature central nervous system; these compounds also had potent inhibitory effects on cultures enriched for brain cancer stem cells. These results suggest that clinically approved neuromodulators may remodel the mature central nervous system and find application in the treatment of brain cancer.

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Figure 1: HTS of NPCs.
Figure 2: Identification of potent NPC-specific compounds.
Figure 3: Temporal effects of neuromodulators on NPC viability and apoptotic response.
Figure 4: Neuromodulator drug sensitivity in normal and cancerous NPCs.

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Acknowledgements

We thank L. Lee, P. Northcode and S. Dolma for technical expertise, G. Giaever and C. Nislow for access to cheminformatics software, and S. Cordes for critical suggestions regarding pharmacological rescue experiments. P.D. is supported by a Canadian Institutes of Health Research (CIHR) MD/PhD studentship. P.B.D. is supported by grants from the National Cancer Institute of Canada, with funds from the Canadian Cancer Society, CIHR and the Stem Cell Network, and from donations from BrainChild and the Durigon (Jessica's Footprint), Baker, Blyth and Kolic families. M.T. is supported by grants from the CIHR and a Canada Research Chair in Bioinformatics and Functional Genomics.

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Correspondence to Mike Tyers or Peter B Dirks.

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Supplementary information

Supplementary Fig. 1

Bioactive neuromodulators display a rich intraclass chemical diversity. (PDF 108 kb)

Supplementary Fig. 2

SAR analysis of adenosine and serotonin agonists. (PDF 125 kb)

Supplementary Fig. 3

SAR analysis of dopamine agonists and antagonists. (PDF 64 kb)

Supplementary Table 1

HTS of neural precursor cells. (PDF 97 kb)

Supplementary Table 2

HTS bioactive pharmacological classes. (PDF 9 kb)

Supplementary Table 3

Assessment of the potency and selectivity of a subset of the identified bioactive agents. (PDF 144 kb)

Supplementary Table 4

Normal and cancerous human neural precursor cells show sensitivity to a myriad of neurotransmission modulators. (PDF 129 kb)

Supplementary Methods (PDF 2256 kb)

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Diamandis, P., Wildenhain, J., Clarke, I. et al. Chemical genetics reveals a complex functional ground state of neural stem cells. Nat Chem Biol 3, 268–273 (2007). https://doi.org/10.1038/nchembio873

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