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Principal Investigator: LORENZ P. STUDER
Organization: SLOAN-KETTERING INST CAN RESEARCH
Fiscal Year: 2024
Award: $635,667
Funding agency: National Institute of Neurological Disorders and Stroke
Project Summary
Human embryonic (hESC) and human induced pluripotent stem cells (hiPSC) offer great promise for basic
research and for applications in disease modeling. The initial challenge for exploiting this potential was to direct
stem cell differentiation towards specific nerve cell or glial fates relevant to disease. Over the last few years, we
have developed many such protocols that now enable researchers to generate > 50 distinct human cell types in
a dish. However, a major remaining challenge is the fetal rather than adult-like features exhibited by the
resulting cells, which limits their usefulness. The reason for their immaturity is unclear but may be linked to a
cell-intrinsic, clock-like mechanism that controls the timing of maturation. While it takes 9 months for a human
baby to develop, from conception to birth, the same process takes only 20 days in a mouse. Those dramatic
timing differences are recapitulated in a dish, where the maturation of human cells may require many months
to reach adult-like properties. Interestingly, we observe such timing differences in both 2D and 3D culture
including in neural organoids. Even after transplanting human cells into the mouse brain, cells continue to
follow a human-specific maturation trajectory, despite being surrounded by an adult host microenvironment.
Here we will address this challenge by building assays to measure and quantify neuronal and glial
maturation and by developing strategies to override the intrinsic maturation clock. Towards these goals, we
have established a unique stem cell-based assay to produce nerve cells at very high precision and in a
temporally synchronized manner. The resulting cells then progressively mature from fetal to adult-like stages
over a period of several months allowing us to define markers that predict the neuronal maturation state. In Aim
1, we will build on these preliminary data and establish stage-specific “fingerprints” of nerve cell maturation to
determine maturation states at unprecedented precision. In addition, we will characterize the maturation of glial
cells (astrocytes and microglia) in a novel tri-culture system to test whether the presence of glia can improve
neuronal maturation. In Aim 2, we will apply maturation “fingerprints” as a readout for identifying factors that
can accelerate maturation timing. In preliminary studies, we have identified chemicals and genes that are
strong candidates for driving neuronal maturation. We will further validate those findings in the tri-culture
system to determine the combined effect of intrinsic and extrinsic maturation factors. Finally, we will perform
mechanistic studies to understand how those factors induce more adult-like features in human cells. In Aim3,
we will test our optimized maturation strategies in more complex 3D culture systems to assess whether
induced maturation strategies impact other developmental processes such as cell migration and organization.
Finally, we will assess whether “induced maturation” strategies can be adopted to achieve accelerated
timelines of neuronal maturation upon transplantation into the developing murine brain, a strategy that could
enable new human disease models for the study of neurodevelopmental or neuropsychiatric disorders in vivo.
Terms: <21+ years old><3-D><3-Dimensional><3D><3D cell culture><3D culture><Acceleration><Address><Adopted><Adult><Adult Human><Assay><Astrocytes><Astrocytus><Astroglia><Automobile Driving><Basic Research><Basic Science><Behavior><Bioassay><Biological Assay><Biology><Birth><Brain><Brain Nervous System><Cell Body><Cell Locomotion><Cell Maturation><Cell Migration><Cell Movement><Cells><Cellular Migration><Cellular Motility><Characteristics><Chemicals><Chromatin><Complex><Conceptions><Data><Degenerative Neurologic Disorders><Development><Developmental Process><Disease><Disorder><Embryo><Embryonic><Encephalon><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Exhibits><Fingerprint><Future><Genes><Genetic><Glia><Glial Cells><Goals><Heterograft><Heterologous Transplantation><Hortega cell><Human><Human Characteristics><Human Development><Human Nature><In Vitro><Intrinsic factor><Investigators><Kolliker's reticulum><Link><Measures><Mice><Mice Mammals><Microglia><Modeling><Modern Man><Molecular><Molecular Fingerprinting><Molecular Profiling><Murine><Mus><Nerve Cells><Nerve Unit><Nervous System><Nervous System Degenerative Diseases><Neural Cell><Neural Degenerative Diseases><Neural Stem Cell><Neural degenerative Disorders><Neurocyte><Neurodegenerative Diseases><Neurodegenerative Disorders><Neurodevelopmental Disorder><Neuroglia><Neuroglial Cells><Neurologic Body System><Neurologic Degenerative Conditions><Neurologic Organ System><Neurological Development Disorder><Neuronal Differentiation><Neurons><Non-neuronal cell><Nonneuronal cell><Organoids><Parturition><Process><Progenitor Cells><Property><Protocol><Protocols documentation><Regenerative Medicine><Reporter><Research Personnel><Researchers><Role><Study models><System><Testing><Transplantation><Work><Xenograft><Xenograft procedure><Xenotransplantation><adulthood><astrocytic glia><cell motility><cell type><degenerative diseases of motor and sensory neurons><degenerative neurological diseases><developmental><differentiation of pluripotent stem cells><differentiation protocol><directed differentiation><disease model><disorder model><driving><epigenetically><excitatory neuron><fetal><genetic approach><genetic strategy><gitter cell><hiPSC><human disease><human iPS><human iPSC><human induced pluripotent cell><human induced pluripotent stem cells><human inducible stem cells><human model><human pluripotent stem cell><human progenitor cell derived><human stem cell-derived><improved><in utero transplantation><in vivo><induced human pluripotent stem cells><inhibitory neuron><intra-uterine transplantation><intrauterine transplantation><mesoglia><microglial cell><microgliocyte><model of human><molecular profile><molecular signature><nerve cement><nerve stem cell><neural><neural precursor><neural precursor cell><neural progenitor><neural progenitor cells><neurodegenerative illness><neurodevelopmental disease><neuron development><neuron progenitors><neuronal><neuronal development><neuronal progenitor><neuronal progenitor cells><neuronal stem cells><neuroprogenitor><neuropsychiatric disease><neuropsychiatric disorder><novel><perivascular glial cell><pharmacologic><pluripotent stem cell differentiation><predictive biomarkers><predictive marker><predictive molecular biomarker><prenatal transplantation><prevent><preventing><progenitor cell differentiation><progenitor differentiation><programs><social role><stem><stem and progenitor differentiation><stem cell differentiation><stem cells><three dimensional><three dimensional cell culture><timeline><tool><transplant><xeno-transplant><xeno-transplantation>