Novel human pluripotent stem cell-derived hypothalamus organoids demonstrate cellular diversity
Lily Sarrafha,
Drew R. Neavin,
Gustavo M. Parfitt,
Ilya A. Kruglikov,
Kristen Whitney,
Ricardo Reyes,
Elena Coccia,
Tatyana Kareva,
Camille Goldman,
Regine Tipon,
Gist Croft,
John F. Crary,
Joseph E. Powell,
Joel Blanchard,
Tim Ahfeldt
Affiliations
Lily Sarrafha
Nash Family Department of Neuroscience, Mount Sinai, New York, NY 10029, USA; Department of Neurology, Mount Sinai, New York, NY 10029, USA; Department of Cell, Developmental and Regenerative Biology, Mount Sinai, New York, NY 10029, USA; Ronald M. Loeb Center for Alzheimer’s Disease, Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Mount Sinai, New York, NY 10029, USA; Black Family Stem Cell Institute, Mount Sinai, New York, NY 10029, USA
Drew R. Neavin
Garvan-Weizmann Centre for Cellular Genomics, Garvan Institute of Medical Research, Darlinghurst, Sydney, NSW 2010, Australia
Gustavo M. Parfitt
Nash Family Department of Neuroscience, Mount Sinai, New York, NY 10029, USA; Ronald M. Loeb Center for Alzheimer’s Disease, Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Mount Sinai, New York, NY 10029, USA; Black Family Stem Cell Institute, Mount Sinai, New York, NY 10029, USA
Ilya A. Kruglikov
New York Stem Cell Foundation, New York, NY 10019, USA
Kristen Whitney
Nash Family Department of Neuroscience, Mount Sinai, New York, NY 10029, USA; Ronald M. Loeb Center for Alzheimer’s Disease, Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Mount Sinai, New York, NY 10029, USA; Department of Pathology, Molecular, and Cell-Based Medicine, Mount Sinai, New York, NY 10029, USA
Ricardo Reyes
Nash Family Department of Neuroscience, Mount Sinai, New York, NY 10029, USA; Department of Neurology, Mount Sinai, New York, NY 10029, USA; Department of Cell, Developmental and Regenerative Biology, Mount Sinai, New York, NY 10029, USA; Ronald M. Loeb Center for Alzheimer’s Disease, Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Mount Sinai, New York, NY 10029, USA; Black Family Stem Cell Institute, Mount Sinai, New York, NY 10029, USA
Elena Coccia
Nash Family Department of Neuroscience, Mount Sinai, New York, NY 10029, USA; Department of Neurology, Mount Sinai, New York, NY 10029, USA; Department of Cell, Developmental and Regenerative Biology, Mount Sinai, New York, NY 10029, USA; Ronald M. Loeb Center for Alzheimer’s Disease, Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Mount Sinai, New York, NY 10029, USA; Black Family Stem Cell Institute, Mount Sinai, New York, NY 10029, USA
Tatyana Kareva
Nash Family Department of Neuroscience, Mount Sinai, New York, NY 10029, USA; Department of Neurology, Mount Sinai, New York, NY 10029, USA; Department of Cell, Developmental and Regenerative Biology, Mount Sinai, New York, NY 10029, USA; Ronald M. Loeb Center for Alzheimer’s Disease, Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Mount Sinai, New York, NY 10029, USA; Black Family Stem Cell Institute, Mount Sinai, New York, NY 10029, USA
Camille Goldman
Nash Family Department of Neuroscience, Mount Sinai, New York, NY 10029, USA; Department of Neurology, Mount Sinai, New York, NY 10029, USA; Department of Cell, Developmental and Regenerative Biology, Mount Sinai, New York, NY 10029, USA; Ronald M. Loeb Center for Alzheimer’s Disease, Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Mount Sinai, New York, NY 10029, USA; Black Family Stem Cell Institute, Mount Sinai, New York, NY 10029, USA
Regine Tipon
New York Stem Cell Foundation, New York, NY 10019, USA
Gist Croft
New York Stem Cell Foundation, New York, NY 10019, USA
John F. Crary
Nash Family Department of Neuroscience, Mount Sinai, New York, NY 10029, USA; Ronald M. Loeb Center for Alzheimer’s Disease, Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Mount Sinai, New York, NY 10029, USA; Department of Pathology, Molecular, and Cell-Based Medicine, Mount Sinai, New York, NY 10029, USA; Windreich Department of Artificial Intelligence and Human Health, Mount Sinai, New York, NY 10029, USA
Joseph E. Powell
Garvan-Weizmann Centre for Cellular Genomics, Garvan Institute of Medical Research, Darlinghurst, Sydney, NSW 2010, Australia; UNSW Cellular Genomics Futures Institute, University of New South Wales, Kensington, Sydney, NSW 2052, Australia
Joel Blanchard
Nash Family Department of Neuroscience, Mount Sinai, New York, NY 10029, USA; Department of Cell, Developmental and Regenerative Biology, Mount Sinai, New York, NY 10029, USA; Ronald M. Loeb Center for Alzheimer’s Disease, Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Mount Sinai, New York, NY 10029, USA; Black Family Stem Cell Institute, Mount Sinai, New York, NY 10029, USA; Corresponding author
Tim Ahfeldt
Nash Family Department of Neuroscience, Mount Sinai, New York, NY 10029, USA; Department of Neurology, Mount Sinai, New York, NY 10029, USA; Department of Cell, Developmental and Regenerative Biology, Mount Sinai, New York, NY 10029, USA; Ronald M. Loeb Center for Alzheimer’s Disease, Mount Sinai, New York, NY 10029, USA; Friedman Brain Institute, Mount Sinai, New York, NY 10029, USA; Black Family Stem Cell Institute, Mount Sinai, New York, NY 10029, USA; Corresponding author
Summary: The hypothalamus is a region of the brain that plays an important role in regulating body functions and behaviors. There is a growing interest in human pluripotent stem cells (hPSCs) for modeling diseases that affect the hypothalamus. Here, we established an hPSC-derived hypothalamus organoid differentiation protocol to model the cellular diversity of this brain region. Using an hPSC line with a tyrosine hydroxylase (TH)-TdTomato reporter for dopaminergic neurons (DNs) and other TH-expressing cells, we interrogated DN-specific pathways and functions in electrophysiologically active hypothalamus organoids. Single-cell RNA sequencing (scRNA-seq) revealed diverse neuronal and non-neuronal cell types in mature hypothalamus organoids. We identified several molecularly distinct hypothalamic DN subtypes that demonstrated different developmental maturities. Our in vitro 3D hypothalamus differentiation protocol can be used to study the development of this critical brain structure and can be applied to disease modeling to generate novel therapeutic approaches for disorders centered around the hypothalamus.
