Tourette Syndrome (TS) is a disorder of the developing telencephalon for which no significant causative genetic variant has yet emerged through the examination of blood samples. In this proposal we investigate whether somatic mutations might underlie in part the pathogenesis of TS. Existing evidence suggests that cells accumulate somatic mutations after the formation of the zygote, implying that cells of the human body do not have identical DNA sequence. Besides single nucleotide variation (SNV) and small insertion/deletions (InDels), cells can accumulate copy number variations (CNVs, i.e., duplications and deletions), insertions of transposable elements, inversions and translocations, all involving from few hundred to several millions of nucleotides. Somatic mosaicism arising in brain cells could explain the failure to discover consistent, replicable genetic risk factors in neuropsychiatric disorders like TS, and underlie at least in part the frequently observed variability between blood genotype and overall phenotype. There is no estimate of somatic mosaicism in either normal development or in disease. To test the hypothesis that somatic mutations might underlie the emergence of TS, in this proposal we will discover and quantify somatic genome variation in TS and normal control brains, followed by exploration of potential functional consequences of this variation.
In Aim 1, we will perform using advanced sequencing techniques comprehensive discovery of lineage-specific and region-specific somatic genomic variations: SNVs, InDels, CNVs, retrotransposon insertions, inversion and translocations. The analysis will involve 20 TS brains and matched 20 normal control brains. Mosaic variants will be discovered and validated in prefrontal cortex (PFC), premotor cortex (PMC) and striatum (STR), three regions strongly implicated in TS, as well as in specific cell lineages isolated from these regions, including pyramidal neurons, medium spiny neurons, interneurons and microglial cells.
In Aim 2, we will select 10 genomic variants, engineer them into iPSCs and in transgenic mice using CRISPR technologies, and characterize their impact on the molecular, tissue and behavior level. Together, these specific aims will provide the first estimate of somatic genomic variation (number, type, frequency) in the brain of TS and will yield hypotheses about their significance for brain development.

Public Health Relevance

New evidence suggests that cells of the brain acquire mutations during development, implying that the brain is a mosaic of cells with different DNA sequence, a phenomenon called somatic mosaicism. Using postmortem brain tissue, in this proposal we investigate the extent of somatic mosaicism in cells of the cerebral cortex and striatum of individual with Tourette Syndrome (TS) and matched controls. We will then explore the functional consequences of somatic mutations by expressing them in mouse models and in human induced pluripotent stem cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01MH106876-04
Application #
9414083
Study Section
Special Emphasis Panel (ZMH1)
Program Officer
Senthil, Geetha
Project Start
2015-05-01
Project End
2020-01-31
Budget Start
2018-02-01
Budget End
2019-01-31
Support Year
4
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Yale University
Department
Psychiatry
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520