Behavioral changes and growth deficits in a crispr engineered mouse model of the schizophrenia-associated 3q29 deletion

ABSTRACT The 3q29 deletion confers increased risk for neuropsychiatric phenotypes including intellectual disability, autism spectrum disorder, generalized anxiety disorder, and a >40-fold

increased risk for schizophrenia. To investigate consequences of the 3q29 deletion in an experimental system, we used CRISPR/Cas9 technology to introduce a heterozygous deletion into the

syntenic interval on C57BL/6 mouse chromosome 16. mRNA abundance for 20 of the 21 genes in the interval was reduced by ~50%, while protein levels were reduced for only a subset of these,

suggesting a compensatory mechanism. Mice harboring the deletion manifested behavioral impairments in multiple domains including social interaction, cognitive function, acoustic startle, and

amphetamine sensitivity, with some sex-dependent manifestations. In addition, 3q29 deletion mice showed reduced body weight throughout development consistent with the phenotype of 3q29

deletion syndrome patients. Of the genes within the interval, _DLG1_ has been hypothesized as a contributor to the neuropsychiatric phenotypes. However, we show that _Dlg1__+/-_ mice did not

exhibit the behavioral deficits seen in mice harboring the full 3q29 deletion. These data demonstrate the following: the 3q29 deletion mice are a valuable experimental system that can be

used to interrogate the biology of 3q29 deletion syndrome; behavioral manifestations of the 3q29 deletion may have sex-dependent effects; and mouse-specific behavior phenotypes associated

with the 3q29 deletion are not solely due to haploinsufficiency of _Dlg1_. Access through your institution Buy or subscribe This is a preview of subscription content, access via your

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PSYCHOTIC ILLNESS IN PWS AND SCHIZOPHRENIA Article Open access 20 August 2021 A GENETICS-FIRST APPROACH TO UNDERSTANDING AUTISM AND SCHIZOPHRENIA SPECTRUM DISORDERS: THE 22Q11.2 DELETION

SYNDROME Article Open access 03 October 2022 DISSECTING 16P11.2 HEMI-DELETION TO STUDY SEX-SPECIFIC STRIATAL PHENOTYPES OF NEURODEVELOPMENTAL DISORDERS Article 26 January 2024 REFERENCES *

Ballif BC, Theisen A, Coppinger J, Gowans GC, Hersh JH, Madan-Khetarpal S, et al. Expanding the clinical phenotype of the 3q29 microdeletion syndrome and characterization of the reciprocal

microduplication. Mol Cytogenet. 2008;1:8. Article  Google Scholar  * Glassford MR, Rosenfeld JA, Freedman AA, Zwick ME, Mulle JG, Unique Rare Chromosome Disorder Support G. Novel features

of 3q29 deletion syndrome: results from the 3q29 registry. Am J Med Genet A. 2016;170A:999–1006. Article  Google Scholar  * Marshall CR, Howrigan DP, Merico D, Thiruvahindrapuram B, Wu W,

Greer DS, et al. Contribution of copy number variants to schizophrenia from a genome-wide study of 41,321 subjects. Nat Genet. 2017;49:27–35. Article  CAS  Google Scholar  * Mulle JG, Dodd

AF, McGrath JA, Wolyniec PS, Mitchell AA, Shetty AC, et al. Microdeletions of 3q29 confer high risk for schizophrenia. Am J Hum Genet. 2010;87:229–36. Article  CAS  Google Scholar  * Mulle

JG, Gambello MJ, Cook EH, Rutkowski TP, Glassford M. 3q29 recurrent deletion. In: Adam MP, Ardinger HH, Pagon RA, Wallace SE, Bean LJH, Stephens K, et al., editors. GeneReviews((R)).

Seattle, WA; University of Washington, Seattle; 1993–2019. Available from: https://www.ncbi.nlm.nih.gov/books/NBK385289/. * Sanders SJ, He X, Willsey AJ, Ercan-Sencicek AG, Samocha KE, Cicek

AE, et al. Insights into autism spectrum disorder genomic architecture and biology from 71 risk loci. Neuron. 2015;87:1215–33. Article  CAS  Google Scholar  * Willatt L, Cox J, Barber J,

Cabanas ED, Collins A, Donnai D, et al. 3q29 microdeletion syndrome: clinical and molecular characterization of a new syndrome. Am J Hum Genet. 2005;77:154–60. Article  CAS  Google Scholar 

* Grozeva D, Conrad DF, Barnes CP, Hurles M, Owen MJ, O’Donovan MC, et al. Independent estimation of the frequency of rare CNVs in the UK population confirms their role in schizophrenia.

Schizophr Res. 2012;135:1–7. Article  Google Scholar  * Mulle JG. The 3q29 deletion confers >40-fold increase in risk for schizophrenia. Mol Psychiatry. 2015;20:1028–9. Article  CAS 

Google Scholar  * Korablev AN, Serova IA, Serov OL. Generation of megabase-scale deletions, inversions and duplications involving the Contactin-6 gene in mice by CRISPR/Cas9 technology. BMC

Genet. 2017;18(Suppl 1):112. Article  Google Scholar  * Carroll LS, Williams HJ, Walters J, Kirov G, O’Donovan MC, Owen MJ. Mutation screening of the 3q29 microdeletion syndrome candidate

genes DLG1 and PAK2 in schizophrenia. Am J Med Genet B Neuropsychiatr Genet. 2011;156B:844–9. Article  CAS  Google Scholar  * Rutkowski TP, Schroeder JP, Gafford GM, Warren ST, Weinshenker

D, Caspary T, et al. Unraveling the genetic architecture of copy number variants associated with schizophrenia and other neuropsychiatric disorders. J Neurosci Res. 2017;95:1144–60. Article

  CAS  Google Scholar  * Leonard AS, Davare MA, Horne MC, Garner CC, Hell JW. SAP97 is associated with the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor GluR1 subunit. J

Biol Chem. 1998;273:19518–24. Article  CAS  Google Scholar  * Purcell SM, Moran JL, Fromer M, Ruderfer D, Solovieff N, Roussos P, et al. A polygenic burden of rare disruptive mutations in

schizophrenia. Nature. 2014;506:185–90. Article  CAS  Google Scholar  * Uezato A, Kimura-Sato J, Yamamoto N, Iijima Y, Kunugi H, Nishikawa T. Further evidence for a male-selective genetic

association of synapse-associated protein 97 (SAP97) gene with schizophrenia. Behav Brain Funct. 2012;8:2. Article  CAS  Google Scholar  * Toyooka K, Iritani S, Makifuchi T, Shirakawa O,

Kitamura N, Maeda K, et al. Selective reduction of a PDZ protein, SAP-97, in the prefrontal cortex of patients with chronic schizophrenia. J Neurochem. 2002;83:797–806. Article  CAS  Google

Scholar  * Gupta P, Uner OE, Nayak S, Grant GR, Kalb RG. SAP97 regulates behavior and expression of schizophrenia risk enriched gene sets in mouse hippocampus. PLoS ONE. 2018;13:e0200477.

Article  Google Scholar  * Wang Y, Zeng C, Li J, Zhou Z, Ju X, Xia S, et al. PAK2 haploinsufficiency results in synaptic cytoskeleton impairment and autism-related behavior. Cell Rep.

2018;24:2029–41. Article  CAS  Google Scholar  * Grice SJ, Liu JL, Webber C. Synergistic interactions between _Drosophila_ orthologues of genes spanned by de novo human CNVs support

multiple-hit models of autism. PLoS Genet. 2015;11:e1004998. Article  Google Scholar  * Mahoney ZX, Sammut B, Xavier RJ, Cunningham J, Go G, Brim KL, et al. Discs-large homolog 1 regulates

smooth muscle orientation in the mouse ureter. Proc Natl Acad Sci USA. 2006;103:19872–7. Article  CAS  Google Scholar  * Yang M, Silverman JL, Crawley JN. Automated three-chambered social

approach task for mice. Curr Protoc Neurosci. 2011;Chapter 8: Unit 8 26. * Chalermpalanupap T, Schroeder JP, Rorabaugh JM, Liles LC, Lah JJ, Levey AI, et al. Locus coeruleus ablation

exacerbates cognitive deficits, neuropathology, and lethality in P301S tau transgenic mice. J Neurosci. 2018;38:74–92. Article  CAS  Google Scholar  * Weinshenker D, Miller NS, Blizinsky K,

Laughlin ML, Palmiter RD. Mice with chronic norepinephrine deficiency resemble amphetamine-sensitized animals. Proc Natl Acad Sci USA. 2002;99:13873–7. Article  CAS  Google Scholar  *

R-CoreTeam. A language and environment for statistical computing. Vienna, Austria: R foundation for Statistical Computing; 2017. http://www.R-project.org. * Bates D, Machler M, Bolker BM,

Walker SC. Fitting linear mixed-effects models using lme4. J Stat Softw. 2015;67:1–48. Article  Google Scholar  * Kuznetsova A, Brockhoff PB, Christensen RHB. lmerTest package: tests in

linear mixed effects models. J Stat Softw. 2017;82:1–26. Article  Google Scholar  * Cox DM, Butler MG. A clinical case report and literature review of the 3q29 microdeletion syndrome. Clin

Dysmorphol. 2015;24:89–94. Article  Google Scholar  * Nielsen J, Fejgin K, Sotty F, Nielsen V, Mork A, Christoffersen CT, et al. A mouse model of the schizophrenia-associated 1q21.1

microdeletion syndrome exhibits altered mesolimbic dopamine transmission. Transl Psychiatry. 2017;7:1261. Article  Google Scholar  * Takahashi H, Nakamura T, Kim J, Kikuchi H, Nakahachi T,

Ishitobi M, et al. Acoustic hyper-reactivity and negatively skewed locomotor activity in children with autism spectrum disorders: an exploratory study. Front Psychiatry. 2018;9:355. Article

  Google Scholar  * Kesby JP, Eyles DW, McGrath JJ, Scott JG. Dopamine, psychosis and schizophrenia: the widening gap between basic and clinical neuroscience. Transl Psychiatry. 2018;8:30.

Article  CAS  Google Scholar  * Paval D. A dopamine hypothesis of autism spectrum disorder. Dev Neurosci. 2017;39:355–60. Article  CAS  Google Scholar  * Kidd JM, Cooper GM, Donahue WF,

Hayden HS, Sampas N, Graves T, et al. Mapping and sequencing of structural variation from eight human genomes. Nature. 2008;453:56–64. Article  CAS  Google Scholar  * Logue SF, Paylor R,

Wehner JM. Hippocampal lesions cause learning deficits in inbred mice in the Morris water maze and conditioned-fear task. Behav Neurosci. 1997;111:104–13. Article  CAS  Google Scholar  *

Morris RG, Hagan JJ, Rawlins JN. Allocentric spatial learning by hippocampectomised rats: a further test of the “spatial mapping” and “working memory” theories of hippocampal function. Q J

Exp Psychol B. 1986;38:365–95. CAS  PubMed  Google Scholar  * Baez-Mendoza R, Schultz W. The role of the striatum in social behavior. Front Neurosci. 2013;7:233. Article  Google Scholar  *

Koch M. The neurobiology of startle. Prog Neurobiol. 1999;59:107–28. Article  CAS  Google Scholar  * Murphy MM, Lindsey Burrell T, Cubells JF, Espana RA, Gambello MJ, Goines KCB, et al.

Study protocol for The Emory 3q29 Project: evaluation of neurodevelopmental, psychiatric, and medical symptoms in 3q29 deletion syndrome. BMC Psychiatry. 2018;18:183. Article  Google Scholar

  * Halladay AK, Bishop S, Constantino JN, Daniels AM, Koenig K, Palmer K, et al. Sex and gender differences in autism spectrum disorder: summarizing evidence gaps and identifying emerging

areas of priority. Mol Autism. 2015;6:36. Article  Google Scholar  * Kirkovski M, Enticott PG, Fitzgerald PB. A review of the role of female gender in autism spectrum disorders. J Autism Dev

Disord. 2013;43:2584–603. Article  Google Scholar  * Lai MC, Lombardo MV, Baron-Cohen S. Autism. Lancet. 2014;383:896–910. Article  Google Scholar  * Prendergast BJ, Onishi KG, Zucker I.

Female mice liberated for inclusion in neuroscience and biomedical research. Neurosci Biobehav Rev. 2014;40:1–5. Article  Google Scholar  Download references ACKNOWLEDGEMENTS The work was

supported by R01GM097331 (TC, DW, and STW), R56MH116994 (TC, JGM, DW, and STW) and R01MH110701 along with funds from the Department of Human Genetics at Emory. This study was supported in

part by the Mouse Transgenic and Gene Targeting Core (TMF) and the Rodent Behavioral Core, which are subsidized by the Emory University School of Medicine and are part of the Emory

Integrated Core Facilities. Additional support was provided by the Georgia Clinical and Translational Science Alliance of the National Institutes of Health under Award Number UL1TR002378.

The content is solely the responsibility of the authors and does not necessarily reflect the official views of the National Institutes of Health. We are grateful to Dr. Jeffrey Miner,

Washington University in St Louis, for providing _Dlg1_ mutant mice. This study was also supported by the Emory Winship Research Pathology Core Lab. AUTHOR CONTRIBUTIONS GJB, TC, JGM, STW,

and DW designed the research. TPR, RHP, and RMP performed research with help from GMG, SMG, UAK, RMP, TM, and JPS. MPE, TPR, and RMP analyzed data. TC, JGM, TPR, and DW wrote the manuscript.

All authors provided edits and approved final manuscript. AUTHOR INFORMATION Author notes * These authors contributed equally: David Weinshenker, Tamara Caspary, Jennifer Gladys Mulle

AUTHORS AND AFFILIATIONS * Department of Human Genetics, Emory University School of Medicine, Atlanta, GA, 30322, USA Timothy P. Rutkowski, Rebecca M. Pollak, Stephanie M. Grewenow, 

Georgette M. Gafford, Tamika Malone, Uswa A. Khan, Jason P. Schroeder, Michael P. Epstein, Stephen T. Warren, David Weinshenker, Tamara Caspary & Jennifer Gladys Mulle * Department of

Cell Biology, Emory University School of Medicine, Atlanta, GA, 30322, USA Ryan H. Purcell & Gary J. Bassell Authors * Timothy P. Rutkowski View author publications You can also search

for this author inPubMed Google Scholar * Ryan H. Purcell View author publications You can also search for this author inPubMed Google Scholar * Rebecca M. Pollak View author publications

You can also search for this author inPubMed Google Scholar * Stephanie M. Grewenow View author publications You can also search for this author inPubMed Google Scholar * Georgette M.

Gafford View author publications You can also search for this author inPubMed Google Scholar * Tamika Malone View author publications You can also search for this author inPubMed Google

Scholar * Uswa A. Khan View author publications You can also search for this author inPubMed Google Scholar * Jason P. Schroeder View author publications You can also search for this author

inPubMed Google Scholar * Michael P. Epstein View author publications You can also search for this author inPubMed Google Scholar * Gary J. Bassell View author publications You can also

search for this author inPubMed Google Scholar * Stephen T. Warren View author publications You can also search for this author inPubMed Google Scholar * David Weinshenker View author

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Gladys Mulle View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Jennifer Gladys Mulle. ETHICS DECLARATIONS CONFLICT

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ARTICLE Rutkowski, T.P., Purcell, R.H., Pollak, R.M. _et al._ Behavioral changes and growth deficits in a CRISPR engineered mouse model of the schizophrenia-associated 3q29 deletion. _Mol

Psychiatry_ 26, 772–783 (2021). https://doi.org/10.1038/s41380-019-0413-5 Download citation * Received: 26 November 2018 * Revised: 13 February 2019 * Accepted: 18 March 2019 * Published: 11

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