The recent discovery that de novo zygotic mutations in the form of CNVs and point mutations make major contributions to neuropsychiatric diseases such as schizophrenia and autism begs the question as to the degree to which de novo post-zygotic mosaic mutations also contribute to disease. In this model, a mutation that occurs post-zygotically can seed some percentage of cells in the brain, and is sufficient to lead to neuronal dysfunction and disease. The approach of sequencing only non-neural tissues such as blood may underpower the detection of mosaicism, because mutations may be restricted to neural tissue. This proposal brings together a highly productive and collaborative team, in which each member contributes a special resource to make this effort truly unique. Gleeson and Mathern recently identified among the first de novo somatic mutations in the developing brain in the condition known as `hemimegalencephaly' (HME), a catastrophic neuropsychiatric condition associated with focal cortical disorganization (FCD). By comparing DNA from diseased brain vs. blood, we identified de novo somatic mutations in PIK3CA, AKT3 and MTOR, part of the mTOR pathway, in as few as 8% of brain cells, resulting in perturbations in an entire cerebral hemisphere. Courchesne and Roy recently identified focal patches of abnormal laminar cytoarchitecture in frontal and temporal cortex in the majority of available brain samples of children studied with autism (ASD), and we suggest focal patches akin to FCD may have similar mutations and contribute to disease. The goal of this application is to extend the discovery of mosaicism in patients with epilepsy and autism in which neurohistopathological evidence points to FCD, by sequencing dysplasias compared with adjacent normal tissue and/or blood at the DNA and RNA level at the single-cell level.
We aim to uncover key sets of genes, in which specific de novo mutations, in specific locations, at specific mosaicism levels, is sufficient to produce clinically defined disease. We will combine next-generation sequencing of FCD brain from patients with neuropsychiatric disease with advanced bioinformatics, single-cell sequencing, complete clinical correlated neuroanatomy and mouse modeling. We will: 1] Test for de novo somatic mutations in a retrospective and prospective cohort of FCD presenting with autism or epilepsy. 2] Correlate genetic disease burden with clinical, imaging, histopathological and single-cell sequencing findings. 3] Test mechanisms by which uncovered de novo mutations alter progenitor cell functions in mammalian cerebral cortex.

Public Health Relevance

Focal cortical disorganization (FCD), in which histopathologically diseased brain is adjacent to normal brain, is well documented to associate with neuropsychiatric disease including epilepsy and autism. The only established treatment is cortical resection to remove diseased brain, which can resolve symptoms. Our recent work identified somatic mutations in the mTOR pathway in patients with FCD, and linked FCD to autism. We propose that mutations exist in autism and epilepsy within patches of FCD. We will expand our mutation analysis and study the genetic basis of disease, with the goal of improved therapy.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Project--Cooperative Agreements (U01)
Project #
3U01MH108898-02S1
Application #
9249183
Study Section
Special Emphasis Panel (ZMH1 (04))
Program Officer
Senthil, Geetha
Project Start
2015-09-22
Project End
2020-06-30
Budget Start
2016-08-12
Budget End
2017-06-30
Support Year
2
Fiscal Year
2016
Total Cost
$220,350
Indirect Cost
$90,350
Name
Rockefeller University
Department
Pediatrics
Type
Other Domestic Higher Education
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065
Sorrells, Shawn F; Paredes, Mercedes F; Cebrian-Silla, Arantxa et al. (2018) Human hippocampal neurogenesis drops sharply in children to undetectable levels in adults. Nature 555:377-381
Courchesne, Eric; Pramparo, Tiziano; Gazestani, Vahid H et al. (2018) The ASD Living Biology: from cell proliferation to clinical phenotype. Mol Psychiatry :
McConnell, Michael J; Moran, John V; Abyzov, Alexej et al. (2017) Intersection of diverse neuronal genomes and neuropsychiatric disease: The Brain Somatic Mosaicism Network. Science 356:
Dandekar, Sugandha; Wijesuriya, Hemani; Geiger, Tim et al. (2016) Shared HLA Class I and II Alleles and Clonally Restricted Public and Private Brain-Infiltrating ?? T Cells in a Cohort of Rasmussen Encephalitis Surgery Patients. Front Immunol 7:608
Baek, Seung Tae; Copeland, Brett; Yun, Eun-Jin et al. (2015) An AKT3-FOXG1-reelin network underlies defective migration in human focal malformations of cortical development. Nat Med 21:1445-54