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

doi:10.1016/j.isci.2023.107525

Novel human pluripotent stem cell-derived hypothalamus organoids demonstrate cellular diversity

iScience. 2023 Aug 2;26(9):107525. doi: 10.1016/j.isci.2023.107525. eCollection 2023 Sep 15.

Authors

Lily Sarrafha^{1

2

3

4

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6}, Drew R Neavin⁷, Gustavo M Parfitt^{1

4

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6}, Ilya A Kruglikov⁸, Kristen Whitney^{1

4

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9}, Ricardo Reyes^{1

2

3

4

5

6}, Elena Coccia^{1

2

3

4

5

6}, Tatyana Kareva^{1

2

3

4

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6}, Camille Goldman^{1

2

3

4

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6}, Regine Tipon⁸, Gist Croft⁸, John F Crary^{1

4

5

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10}, Joseph E Powell^{7

11}, Joel Blanchard^{1

3

4

5

6}, Tim Ahfeldt^{1

2

3

4

5

6}

Affiliations

¹ Nash Family Department of Neuroscience, Mount Sinai, New York, NY 10029, USA.
² Department of Neurology, Mount Sinai, New York, NY 10029, USA.
³ Department of Cell, Developmental and Regenerative Biology, Mount Sinai, New York, NY 10029, USA.
⁴ Ronald M. Loeb Center for Alzheimer's Disease, Mount Sinai, New York, NY 10029, USA.
⁵ Friedman Brain Institute, Mount Sinai, New York, NY 10029, USA.
⁶ Black Family Stem Cell Institute, Mount Sinai, New York, NY 10029, USA.
⁷ Garvan-Weizmann Centre for Cellular Genomics, Garvan Institute of Medical Research, Darlinghurst, Sydney, NSW 2010, Australia.
⁸ New York Stem Cell Foundation, New York, NY 10019, USA.
⁹ Department of Pathology, Molecular, and Cell-Based Medicine, Mount Sinai, New York, NY 10029, USA.
¹⁰ Windreich Department of Artificial Intelligence and Human Health, Mount Sinai, New York, NY 10029, USA.
¹¹ UNSW Cellular Genomics Futures Institute, University of New South Wales, Kensington, Sydney, NSW 2052, Australia.

Abstract

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.

Keywords: Cell biology; Neuroscience; Stem cells research; Transcriptomics.

Grants and funding

T32 AG049688/AG/NIA NIH HHS/United States