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Principal Investigator: MIRJANA MALETIC-SAVATIC
Organization: BOSTON CHILDREN'S HOSPITAL
Fiscal Year: 2024
Award: $832,810
Funding agency: National Institute of Mental Health
Project Summary/Abstract
Copy number variations (CNVs) of the human 16p11.2 genetic locus, containing 29 coding genes, are associated
with a number of neurodevelopmental and psychiatric disorders. The deletion (16pdel) and duplication (16pdup)
variants of this region have poorly understood pleiotropic effects. Although autism is more common in patients
with deletions, and schizophrenia is more common in those with duplications, underlying mechanisms are not
clear. Several molecular pathways from the 16p11.2 region modulate neuronal differentiation, migration, axonal
development, and synapse formation, as well as energy and lipid metabolism. Studies of 16p-animal models
have suggested deficits in the KCTD13-RhoA pathway activation, neuronal migration, axonal development, and
behavior. In turn, disruption of ceramide homeostasis due to 16p11.2 CNVs at FAM57B locus altered lipid
abundance, cell membrane dynamics, synaptic protein expression, and synaptic transport, suggesting that
lipidome dysregulation could contribute to neuronal function and activity. However, the exact molecular
mechanisms underlying these neuronal dysfunctions in excitatory versus inhibitory neurons are lacking.
Moreover, contradictory results have been reported from different animal and human cell models. To address
this gap of knowledge, we developed human iPSC-derived neuronal models of 16p11.2 CNVs and demonstrated
that i) KCTD13 regulates RhoA pathway activation, and increased RhoA expression leads to hyperactivity of the
16pdel human cortical neuron networks, and ii) there are significant changes in key mitochondrial and lipid
enzyme transcripts, including decrease in FAM57B-mediated ceramide synthase expression, that directly
correlate with observed changes in the metabolome and lipidome. These data suggest that 16pdel leads to
complex metabolic disruptions and deficient ceramide expression that might contribute to the observed functional
neuronal network phenotypes. These data have led us to hypothesize that 16p11.2 CNVs cause dysregulation
of ceramide abundance in glutamatergic and GABAergic neurons that in turn promotes deficits in synaptic
development and function leading to network disorganization and hyperactivation. Here, we will investigate this
hypothesis and the effects of 16p11.2 CNVs on cortical neuron development and function in human iPSC-derived
2-dimensional excitatory-inhibitory neuron co-cultures and human iPSC-derived 3-dimensional forebrain
organoids. To reduce variability caused by different genotypic backgrounds, we will study CRISPR-Cas9 induced
16p11.2 CNV iPSC lines in addition to iPSC lines derived from patients and healthy controls. We will utilize state-
of-the-art molecular methodologies to uncover mechanisms underlying the synaptic dysfunction of the excitatory-
inhibitory neurons in 16p11.2 CNVs, including single cell transcriptional gene expression profiling and
lipidome/metabolome profiling. Finally, we will investigate the excitatory-inhibitory network function, connectivity,
and oscillation patterns with multi-electrode arrays and patch clamping. We anticipate that this study will uncover
new molecular targets related to cortical neuron dysfunction in 16p11.2 CNV disorders.
Terms: <16p><16p11.2><2-dimensional><3-D><3-D modeling><3-Dimensional><3D><3D modeling><ASD><Address><Affect><Animal Model><Animal Models and Related Studies><Animals><Architecture><Autism><Autistic Disorder><Autoregulation><Axon><Basal Transcription Factor><Basal transcription factor genes><Behavior><Bioenergetics><Bipolar Affective Psychosis><Bipolar Disorder><Brain><Brain Diseases><Brain Disorders><Brain Nervous System><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cas nuclease technology><Cell Body><Cell Communication and Signaling><Cell Line><Cell Signaling><Cell membrane><Cell model><CellLine><Cells><Cellular Matrix><Cellular model><Ceramides><Chromosome 16 Proximal Arm><Chromosome 16 Short Arm><Clinical><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Co-culture><Cocultivation><Coculture><Coculture Techniques><Code><Coding System><Complex><Copy Number Polymorphism><Cytoplasmic Membrane><Cytoskeletal System><Cytoskeleton><DA Neuron><Data><Development><Disease><Disorder><Dopamine neuron><Dysfunction><Early Infantile Autism><Encephalon><Encephalon Diseases><Energy Expenditure><Energy Metabolism><Engineering><Engineering / Architecture><Environment><Enzyme Gene><Enzymes><Fore-Brain><Forebrain><Functional disorder><Gene Expression Monitoring><Gene Expression Pattern Analysis><Gene Expression Profiling><Gene Transcription><General Transcription Factor Gene><General Transcription Factors><Generations><Genes><Genetic Transcription><Genotype><Glutamates><Homeostasis><Human><Hyperactivity><Induced pluripotent stem cell derived human neuron><Induced pluripotent stem cell derived neurons><Infantile Autism><Intellectual disability><Intellectual functioning disability><Intellectual limitation><Intermediary Metabolism><Intracellular Communication and Signaling><Intracranial CNS Disorders><Intracranial Central Nervous System Disorders><Kanner's Syndrome><Knowledge><L-Glutamate><Laboratories><Lipids><Manic-Depressive Psychosis><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Mediating><Mental disorders><Mental health disorders><Mesencephalon><Metabolic><Metabolic Processes><Metabolism><Methodology><Mid-brain><Midbrain><Midbrain structure><Mitochondria><Modeling><Modern Man><Molecular><Molecular Target><Nerve Cells><Nerve Unit><Neural Cell><Neurites><Neurocognitive><Neurocyte><Neurodevelopmental Disorder><Neurological Development Disorder><Neuron from iPSC><Neuron from induced pluripotent stem cells><Neuronal Differentiation><Neuronal Dysfunction><Neurons><Organoids><Pathology><Pathway interactions><Patients><Pattern><Penetrance><Phenotype><Physiological Homeostasis><Physiopathology><Plasma Membrane><Property><Prosencephalon><Psychiatric Disease><Psychiatric Disorder><RNA Expression><RNA Seq><RNA sequencing><RNAseq><Regulation><Reporting><Risk Factors><Role><Schizophrenia><Schizophrenic Disorders><Signal Transduction><Signal Transduction Systems><Signaling><Sorting><Strains Cell Lines><Synapses><Synaptic><Time><Transcript><Transcript Expression Analyses><Transcript Expression Analysis><Transcription><Transcription Factor Proto-Oncogene><Transcription Regulation><Transcription factor genes><Transcriptional Control><Transcriptional Regulation><Variant><Variation><analyze gene expression><autism spectral disorder><autism spectrum disorder><autistic spectrum disorder><biological signal transduction><bipolar affective disorder><bipolar disease><bipolar illness><bipolar mood disorder><brain volume><ceramide synthase><clinical phenotype><connectome><copy number variant><copy number variation><cultured cell line><dementia praecox><density><developmental><dihydroceramide desaturase><dopaminergic neuron><excitatory neuron><fat metabolism><gene expression analysis><gene expression assay><gene locus><genetic locus><genomic location><genomic locus><global gene expression><global transcription profile><glutamatergic><hiPSC><hiPSC-derived neurons><human iPS><human iPSC><human iPSC-derived sensory neuron><human induced pluripotent cell><human induced pluripotent stem cells><human inducible stem cells><iPS><iPS neurons><iPSC><iPSC derived-neurons><iPSC-derived human neuron><iPSCs><induced human pluripotent stem cells><induced pluripotent cell><induced pluripotent stem cell><induced pluripotent stem cell neurons><induced pluripotent stem cells derived from patients><induced pluripotent stem cells from patients><inducible pluripotent stem cell><inhibitory neuron><insight><intellectual and developmental disability><intracellular skeleton><limited intellectual functioning><lipid metabolism><lipidome><lipidomics><manic depressive disorder><manic depressive illness><mental illness><metabolism measurement><metabolome><metabolomics><metabonome><metabonomics><migration><mitochondrial><model of animal><molecular phenotype><multi-electrode arrays><multi-modal data><multi-modal datasets><multielectrode arrays><multimodal data><multimodal datasets><nerve cell metabolism><neural cell body><neural dysfunction><neurite growth><neurodevelopmental disease><neuron cell metabolism><neuron development><neuron metabolism><neuronal><neuronal cell body><neuronal cell metabolism><neuronal circuit><neuronal circuitry><neuronal development><neuronal excitability><neuronal metabolism><neurons derived from induced pluripotent stem cells><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic target><novel therapy target><patch clamp><pathophysiology><pathway><patient derived human iPS><patient derived human iPSC><patient derived human induced pluripotent stem cell><patient derived iPS><patient derived iPSC><patient derived induced pluripotent cells><patient derived induced pluripotent stem cells><patient-derived pluripotent stem cells><plasmalemma><pleiotropic effect><pleiotropism><pleiotropy><protein expression><psychiatric illness><psychological disorder><schizophrenic><social role><soma><synapse><synapse formation><synapse function><synaptic function><synaptogenesis><three dimensional><three-dimensional modeling><tool><transcription factor><transcriptional profiling><transcriptome><transcriptome sequencing><transcriptomic sequencing><two-dimensional>