Genome-wide In Vivo CNS Screening Identifies Genes that Modify CNS Neuronal Survival and mHTT Toxicity
- Creators
- Wertz, Mary H.
- Mitchem, Mollie R.
- Pineda, S. Sebastian
- Hachigian, Lea J.
- Lee, Hyeseung
- Lau, Vanessa
- Powers, Alex
- Kulicke, Ruth
- Madan, Gurrein K.
- Colic, Medina
- Therrien, Martine
- Vernon, Amanda
- Beja-Glasser, Victoria F.
- Hegde, Mudra
- Gao, Fan
- Kellis, Manolis
- Hart, Traver
- Doench, John G.
- Heiman, Myriam
Abstract
Unbiased in vivo genome-wide genetic screening is a powerful approach to elucidate new molecular mechanisms, but such screening has not been possible to perform in the mammalian central nervous system (CNS). Here, we report the results of the first genome-wide genetic screens in the CNS using both short hairpin RNA (shRNA) and CRISPR libraries. Our screens identify many classes of CNS neuronal essential genes and demonstrate that CNS neurons are particularly sensitive not only to perturbations to synaptic processes but also autophagy, proteostasis, mRNA processing, and mitochondrial function. These results reveal a molecular logic for the common implication of these pathways across multiple neurodegenerative diseases. To further identify disease-relevant genetic modifiers, we applied our screening approach to two mouse models of Huntington's disease (HD). Top mutant huntingtin toxicity modifier genes included several Nme genes and several genes involved in methylation-dependent chromatin silencing and dopamine signaling, results that reveal new HD therapeutic target pathways.
Additional Information
© 2020 Elsevier Inc. Received 22 July 2019, Revised 3 December 2019, Accepted 6 January 2020, Available online 30 January 2020. The authors thank the CHDI Foundation for providing the zQ175 mice used in this study. This work was supported by NIH/NINDS award R01 NS085880 (M. Heiman); an award from the JPB Foundation (M. Heiman); an award from the Bev Hartig Huntington's Disease Foundation (M. Heiman); a Fay/Frank Seed Award from the Brain Research Foundation (M. Heiman); the Jeptha H. and Emily V. Wade Award (M. Heiman); fellowships from the JPB Foundation and the Hereditary Disease Foundation (M.H.W.); the NIH (5T32EB019940-05; S.S.P.); and the JPB Foundation (H.L.). T.H. and M.C. were supported by NIH/NIGMS award R35GM130119 (T.H.), CPRIT grant RR160032 (T.H.), and the NIH/NCI Cancer Center Support Grant P30 CA016672. Author Contributions: M.H.W. performed the in vivo screening and Nme1 experiments and analyses. M.R.M., A.P., and L.J.H. performed the intracranial stereotaxic injections. M.C. and T.H. performed the DrugZ analysis. R.K. conducted immunohistochemical experiments and mouse handling. A.V. assisted in mouse handling. H.L., G.K.M., and V.F.B.-G. assisted in Nme1 experiments. M.T. assisted in Nme1 experiments and data analysis. S.S.P. and H.L. prepared and S.S.P. analyzed the snRNA-seq data of Nme1OX experiments. L.J.H. collected and F.G. analyzed the RNA-seq data. M. Hegde and J.G.D. assisted in screening design and data analysis. M.H.W. and M. Heiman wrote the manuscript. M. Heiman conceived and supervised the project. All authors reviewed and approved the final manuscript. The authors declare no competing interests.Attached Files
Accepted Version - nihms-1569339.pdf
Supplemental Material - 1-s2.0-S0896627320300040-mmc1.pdf
Supplemental Material - 1-s2.0-S0896627320300040-mmc2.xlsx
Supplemental Material - 1-s2.0-S0896627320300040-mmc3.xlsx
Supplemental Material - 1-s2.0-S0896627320300040-mmc4.xlsx
Supplemental Material - 1-s2.0-S0896627320300040-mmc5.xlsx
Supplemental Material - 1-s2.0-S0896627320300040-mmc6.xlsx
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Additional details
- PMCID
- PMC7181458
- Eprint ID
- 101005
- DOI
- 10.1016/j.neuron.2020.01.004
- Resolver ID
- CaltechAUTHORS:20200130-124820336
- R01 NS085880
- NIH
- JPB Foundation
- Bev Hartig Huntington's Disease Foundation
- Brain Research Foundation
- Jeptha H. and Emily V. Wade Award
- Hereditary Disease Foundation
- 5T32EB019940-05
- NIH
- R35GM130119
- NIH
- RR160032
- Cancer Prevention and Research Institute of Texas
- P30 CA016672
- NIH
- Created
-
2020-01-30Created from EPrint's datestamp field
- Updated
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2023-07-20Created from EPrint's last_modified field