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Principal Investigator: Jianping Fu
Organization: UNIVERSITY OF MICHIGAN AT ANN ARBOR
Fiscal Year: 2024
Award: $164,992
Funding agency: National Institute of Neurological Disorders and Stroke
Project Summary
Modeling NDE1 function in dysregulated brain development using a microfluidic CNS model
Development of the vertebrate central nervous system (CNS) begins with the formation of neural tube (NT) and
its regional patterning to generate the forebrain, midbrain, hindbrain, and spinal cord. Regional patterning of
the brain is a tightly regulated developmental process, deviation from which can result in neurodevelopmental
brain diseases. Multiple causes are associated with neurodevelopmental brain diseases, including genetic,
environmental, infectious, and traumatic factors. Even though the precise etiology of neurodevelopmental
brain diseases remains largely unknown, the genetic components of neurodevelopmental brain diseases have
been increasingly deciphered with the advent of personalized medicine. However, detailed pathophysiological
mechanisms of neurodevelopmental brain diseases remain challenging to study, due to limited access to
human CNS tissues. Animal models have been instrumental in understanding human neurodevelopment and
associated disorders. However, they are limited in revealing some of the most fundamental aspects of
development, genetics, pathology, and disease mechanisms that are unique to humans. Stem cell-based in
vitro models of human neurodevelopment are emerging as promising experimental tools. However, the
controllability and reproducibility of these models remain suboptimal. Furthermore, none of the current
neurodevelopment models is capable of recapitulating regional patterning of the brain faithfully in a 3D tubular
geometry, a hallmark of neurodevelopment.
The goal of this R21 project is to specifically address this significant technological need, by using
human pluripotent stem cells (hPSCs) to develop a human brain development model that can faithfully
recapitulate regional brain patterning. Importantly, we propose to apply this model to study the function of
nuclear distribution element 1 (NDE1), a gene implicated in a wide range of neurodevelopmental conditions,
including microcephaly (a small brain), microlissencephaly (a small brain with a simplified gyral pattern), or
microhydranencephaly (a more severe presentation). In our preliminary study, we have successfully leveraged
the developmental potential and self-organizing property of hPSCs in conjunction with innovative microfluidics
to develop the first of its kind, synthetic, fully patterned human NT model. Our preliminary data from brain
organoids generated from NDE1-knockout (KO) hPSCs further show that NDE1-KO brain organoids exhibit
reduced growth and gyrification and furthermore show abnormal brain regionalization. Thus, our preliminary
data suggest a novel and previously unexplored mechanism involving dysregulated brain regionalization in
NDE1-mediated microcephaly. In this proposal, we propose to first extend the microfluidic patterned human
NT model to recapitulate brain regionalization (Aim 1). We will then utilize this controllable human brain
development model and a Nde1-KO mouse model to study the role of NDE1 mutations in dysregulated brain
regionalization (Aim 2).
Terms: <3-D><3-Dimensional><3D><Address><Animal Model><Animal Models and Related Studies><Benchmarking><Best Practice Analysis><Biologic Models><Biological Models><Body Tissues><Brain><Brain Diseases><Brain Disorders><Brain Nervous System><Brain region><CNS Nervous System><Causality><Cell Body><Cell division><Cells><Central Nervous System><DNA Alteration><DNA Sequence Alteration><DNA mutation><Data><Development><Developmental Process><Disease><Disorder><Dorsal><Elements><Embryo><Embryonic><Encephalon><Encephalon Diseases><Etiology><Exhibits><Fore-Brain><Forebrain><Generalized Growth><Genes><Genetic><Genetic Alteration><Genetic Change><Genetic defect><Genetic mutation><Genetic study><Geometry><Gestation><Goals><Growth><Health><Hind Brain><Human><Individual><Intracranial CNS Disorders><Intracranial Central Nervous System Disorders><KO mice><Knock-out><Knock-out Mice><Knockout><Knockout Mice><Measurement><Mediating><Medulla Spinalis><Mesencephalon><Mice><Mice Mammals><Microcephaly><Microfluidic Device><Microfluidic Lab-On-A-Chip><Microfluidic Microchips><Microfluidics><Mid-brain><Midbrain><Midbrain structure><Model System><Modeling><Modern Man><Motility><Murine><Mus><Mutation><Neural Development><Neural Stem Cell><Neural Tube Development><Neural tube><Neuraxis><Nuclear><Null Mouse><Organoids><Pathology><Pattern><Position><Positioning Attribute><Pregnancy><Prevalence><Process><Progenitor Cells><Property><Prosencephalon><Proteins><Protocol><Protocols documentation><Reproducibility><Research><Rhombencephalon><Role><Sequence Alteration><Societies><Spinal Cord><Structure><Technology><Tissue Growth><Tissues><Tubular><Tubular formation><benchmark><brain abnormalities><causation><comparative><developmental><differentiation factors><disease causation><disease model><disease phenotype><disorder model><genome mutation><genome sequencing><genomic alteration><global gene expression><global transcription profile><hindbrain><human model><human pluripotent stem cell><in vitro Model><in vivo><innovate><innovation><innovative><micrencephaly><microencephaly><microfluidic chip><model development><model developments><model of animal><model of human><morphogenic factors><morphogens><mouse model><murine model><mutant><mutant mouse model><nerve stem cell><neural network><neural patterning><neural precursor><neural precursor cell><neural progenitor><neural progenitor cells><neurodevelopment><neuron progenitors><neuronal progenitor><neuronal progenitor cells><neuronal stem cells><neuroprogenitor><novel><ontogeny><personalization of treatment><personalized medicine><personalized therapy><personalized treatment><public health relevance><scRNA-seq><self organization><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><social role><spatiotemporal><stem cells><three dimensional><tool><transcriptome><µfluidic>