hESC-Derived Thalamic Organoids Form Reciprocal Projections When Fused with Cortical Organoids.

Human brain organoid techniques have rapidly advanced to facilitate investigating human brain development and diseases. These efforts have largely focused on generating telencephalon due to its direct relevance in a variety of forebrain disorders. Despite its importance as a relay hub between cortex and peripheral tissues, the investigation of three-dimensional (3D) organoid models for the human thalamus has not been explored. Here, we describe a method to differentiate human embryonic stem cells (hESCs) to thalamic organoids (hThOs) that specifically recapitulate the development of thalamus. Single-cell RNA sequencing revealed a formation of distinct thalamic lineages, which diverge from telencephalic fate. Importantly, we developed a 3D system to create the reciprocal projections between thalamus and cortex by fusing the two distinct region-specific organoids representing the developing thalamus or cortex. Our study provides a platform for understanding human thalamic development and modeling circuit organizations and related disorders in the brain.

[1]  T. Shimogori,et al.  Dynamic spatiotemporal gene expression in embryonic mouse thalamus , 2011, The Journal of comparative neurology.

[2]  Yoshiki Sasai,et al.  Generation of functional hippocampal neurons from self-organizing human embryonic stem cell-derived dorsomedial telencephalic tissue , 2015, Nature Communications.

[3]  K. Toyama,et al.  Laminar specificity of extrinsic cortical connections studied in coculture preparations , 1992, Neuron.

[4]  Jan H Lui,et al.  Non-epithelial stem cells and cortical interneuron production in the human ganglionic eminences , 2013, Nature Neuroscience.

[5]  D. Small,et al.  Proteoglycans in the central nervous system: Role in development, neural repair, and Alzheimer's disease , 2013, IUBMB life.

[6]  Y. Sasai,et al.  Generation of thalamic neurons from mouse embryonic stem cells , 2017, Development.

[7]  D. Geschwind,et al.  Functional cortical neurons and astrocytes from human pluripotent stem cells in 3D culture , 2015, Nature Methods.

[8]  Deanna M. Barch,et al.  Cognition in schizophrenia: core psychological and neural mechanisms , 2012, Trends in Cognitive Sciences.

[9]  Sinisa Hrvatin,et al.  Single-cell transcriptomics of the developing lateral geniculate nucleus reveals insights into circuit assembly and refinement , 2018, Proceedings of the National Academy of Sciences.

[10]  N. Andreasen.  The Role of the Thalamus in Schizophrenia , 1997, Canadian journal of psychiatry. Revue canadienne de psychiatrie.

[11]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[12]  Madeline A. Lancaster,et al.  Cerebral organoids model human brain development and microcephaly , 2013, Nature.

[13]  T. Sejnowski,et al.  Thalamocortical oscillations in the sleeping and aroused brain. , 1993, Science.

[14]  C. Lüscher,et al.  In vivo reprogramming of circuit connectivity in postmitotic neocortical neurons , 2013, Nature Neuroscience.

[15]  M. Tomishima,et al.  Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling , 2009, Nature Biotechnology.

[16]  J. Staiger,et al.  Mapping remodeling of thalamocortical projections in the living reeler mouse brain by diffusion tractography , 2013, Proceedings of the National Academy of Sciences.

[17]  J. Rubenstein,et al.  Subcortical origins of human and monkey neocortical interneurons , 2013, Nature Neuroscience.

[18]  Robert Gentleman,et al.  Using GOstats to test gene lists for GO term association , 2007, Bioinform..

[19]  V. Crunelli,et al.  Childhood absence epilepsy: Genes, channels, neurons and networks , 2002, Nature Reviews Neuroscience.

[20]  Min Tae M Park,et al.  Morphological Alterations in the Thalamus, Striatum, and Pallidum in Autism Spectrum Disorder , 2016, Neuropsychopharmacology.

[21]  C. Hetz,et al.  ER Proteostasis Control of Neuronal Physiology and Synaptic Function , 2018, Trends in Neurosciences.

[22]  S. Quake,et al.  A survey of human brain transcriptome diversity at the single cell level , 2015, Proceedings of the National Academy of Sciences.

[23]  N Yamamoto,et al.  Neural connections between the lateral geniculate nucleus and visual cortex in vitro. , 1989, Science.

[24]  David W. Nauen,et al.  Brain-Region-Specific Organoids Using Mini-bioreactors for Modeling ZIKV Exposure , 2016, Cell.

[25]  M. Eiraku,et al.  Minimization of exogenous signals in ES cell culture induces rostral hypothalamic differentiation , 2008, Proceedings of the National Academy of Sciences.

[26]  Zoltán Molnár,et al.  Thalamocortical development: how are we going to get there? , 2003, Nature Reviews Neuroscience.

[27]  L. Puelles,et al.  Molecular anatomy of the thalamic complex and the underlying transcription factors , 2015, Brain Structure and Function.

[28]  G. Govindaiah,et al.  Fusion of Regionally Specified hPSC-Derived Organoids Models Human Brain Development and Interneuron Migration. , 2017, Cell stem cell.

[29]  Y. Yanagawa,et al.  Ontogeny-recapitulating generation and tissue integration of ES cell–derived Purkinje cells , 2010, Nature Neuroscience.

[30]  Hideshi Kawakami,et al.  Self-organization of polarized cerebellar tissue in 3D culture of human pluripotent stem cells. , 2015, Cell reports.

[31]  Núria Queralt-Rosinach,et al.  DisGeNET: a comprehensive platform integrating information on human disease-associated genes and variants , 2016, Nucleic Acids Res..

[32]  Hans Clevers,et al.  Modeling Development and Disease with Organoids , 2016, Cell.

[33]  Fabian J. Theis,et al.  Diffusion maps for high-dimensional single-cell analysis of differentiation data , 2015, Bioinform..

[34]  Christian Rosenmund,et al.  Vesicular Glutamate Transporter VGLUT2 Expression Levels Control Quantal Size and Neuropathic Pain , 2006, The Journal of Neuroscience.

[35]  R. Guillery,et al.  Functional organization of thalamocortical relays. , 1996, Journal of neurophysiology.

[36]  R. Ramasubbu,et al.  Thalamocortical connectivity in major depressive disorder. , 2017, Journal of affective disorders.

[37]  Juergen A. Knoblich,et al.  Organogenesis in a dish: Modeling development and disease using organoid technologies , 2014, Science.

[38]  Daniel R Weinberger,et al.  Midbrain-like Organoids from Human Pluripotent Stem Cells Contain Functional Dopaminergic and Neuromelanin-Producing Neurons. , 2016, Cell stem cell.

[39]  S. Scholpp,et al.  Building a bridal chamber: development of the thalamus , 2010, Trends in Neurosciences.

[40]  A. Fairén,et al.  Neural cell adhesion molecule, NCAM, regulates thalamocortical axon pathfinding and the organization of the cortical somatosensory representation in mouse , 2012, Front. Mol. Neurosci..

[41]  Jonathan A. Bernstein,et al.  Assembly of functionally integrated human forebrain spheroids , 2017, Nature.

[42]  E. Puelles,et al.  Molecular Regionalization of the Developing Neural Tube , 2012 .

[43]  Joseph G. Gleeson,et al.  Primary Cilia in the Developing and Mature Brain , 2014, Neuron.

[44]  S. Pașca,et al.  The rise of three-dimensional human brain cultures , 2018, Nature.

[45]  Evan Z. Macosko,et al.  Highly Parallel Genome-wide Expression Profiling of Individual Cells Using Nanoliter Droplets , 2015, Cell.

[46]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[47]  Z. Molnár,et al.  Specificity and Plasticity of Thalamocortical Connections in Sema6A Mutant Mice , 2009, PLoS biology.

[48]  J. Bagley,et al.  Fused dorsal-ventral cerebral organoids model complex interactions between diverse brain regions , 2017, Nature methods.