Schizophrenia (SZ) is a devastating brain disorder afflicting ~1% of population. Recent SZ genome-wide association studies (GWAS) have implicated dozens of loci, but causal mechanism for each locus is largely unknown - such knowledge would help to identify novel targets for more effective intervention. One of the most strongly associated SZ loci spans two brain-expressed microRNAs: MIR137 regulates neuronal differentiation, maturation and synaptic function, and about half of SZ GWAS loci contain MIR137 targets, suggesting a central hub role for MIR137 in a SZ gene network. MIR2682 has little known function, but it is predicted by TargetScan6.2 to target ankyrin 3 (ANK3), a gene associated with bipolar disorder in GWAS. To understand the causal mechanism underlying the MIR137 SZ locus, this R21 application will address these three key questions: 1) which of the many equivalently associated SNPs are functional, 2) which genes and in what types of neuron are affected by the risk variants, and 3) whether the risk variants impact SZ-relevant cellular phenotypes. We will leverage both the putatively regulatory common SNPs implicated by GWAS and a rare enhancer SNP identified by our resequencing. We expect the rare enhancer SNP to provide unparalleled insights on the causal mechanism because of its large effect;the risk allele was found to reduce reporter gene expression by >60%. Neurons differentiated from induced pluripotent stem cells (iPSCs) are emerging as a pathophysiologically relevant cellular model for studying brain disorders. We have shown robust expression of MIR137/MIR2682 in such neurons. Our hypothesis is that the risk alleles of both the rare and the common SZ- risk variants reduce expression of MIR137/MIR2682 in iPSC-derived neurons, resulting in SZ-relevant cellular phenotypic changes. We present a rigorous and innovative approach: 1) Instead of simply comparing a few iPSCs from SZ cases vs. controls, which is often underpowered to attribute phenotypic differences to a genetic variant due to variable genetic backgrounds between iPSCs, we will generate isogenic iPSC lines differing only at the SNP of interest by using an efficient transcription activator-like effector nuclease (TALEN)-mediated genome editing. 2) To overcome the heterogeneity of iPSC-derived neurons, we will simultaneously profile in single neurons the expression of different subtype-specific markers as well as of MIR137/MIR2682 and their target genes, and then examine within each subtype of neurons the effect of the SZ risk allele on gene expression. Combining the use of iPSC, genome editing, and single neuron expression profiling, we will pursue two specific aims: (1) To generate isogenic iPSC lines differing only at each of the SZ-risk variants on a unified genetic background, and (2) To determine in isogenic iPSC-derived cortical neurons the effects of SZ-risk alleles on expression of MIR137/MIR2682, and on SZ-relevant basic cellular phenotypes.

Public Health Relevance

We will study the functional effects of schizophrenia genetic risk variants on the expression of the microRNAs, MIR137 and MIR2682, and on schizophrenia-relevant cellular phenotypes in living human neurons. The results will provide causal mechanistic insights into one of the most strongly associated schizophrenia susceptibility loci identified by genome-wide association studies. The created cellular model carrying schizophrenia risk variants will enable further study of the relationship between these variants and schizophrenia-relevant molecular and cellular characteristics, and may also be used in screening for small molecules of therapeutic use targeting MIR137 expression.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21MH102685-01
Application #
8621279
Study Section
Molecular Neurogenetics Study Section (MNG)
Program Officer
Panchision, David M
Project Start
2014-02-15
Project End
2016-01-31
Budget Start
2014-02-15
Budget End
2015-01-31
Support Year
1
Fiscal Year
2014
Total Cost
$195,000
Indirect Cost
$70,000
Name
Northshore University Healthsystem
Department
Type
DUNS #
069490621
City
Evanston
State
IL
Country
United States
Zip Code
60201
Duan, Jubao; Shi, Jianxin; Fiorentino, Alessia et al. (2014) A rare functional noncoding variant at the GWAS-implicated MIR137/MIR2682 locus might confer risk to schizophrenia and bipolar disorder. Am J Hum Genet 95:744-53
Russell, Theron A; Blizinsky, Katherine D; Cobia, Derin J et al. (2014) A sequence variant in human KALRN impairs protein function and coincides with reduced cortical thickness. Nat Commun 5:4858
Shi, Sandra; Leites, Catherine; He, Deli et al. (2014) MicroRNA-9 and microRNA-326 regulate human dopamine D2 receptor expression, and the microRNA-mediated expression regulation is altered by a genetic variant. J Biol Chem 289:13434-44
Shi, Jianxin; Marconett, Crystal N; Duan, Jubao et al. (2014) Characterizing the genetic basis of methylome diversity in histologically normal human lung tissue. Nat Commun 5:3365