Deriving Human Intestinal Organoids with Functional Tissue-Resident Macrophages All From Pluripotent Stem Cells

Overview

Journal Cell Mol Gastroenterol Hepatol

Specialty Gastroenterology

Date 2024 Dec 19

PMID 39701210

Authors

Kentaro Tominaga

Daniel O Kechele

J Guillermo Sanchez

Simon Vales

Ingrid Jurickova

Lizza Roman

Akihiro Asai

Jacob R Enriquez

Heather A McCauley

Keishi Kishimoto

Kentaro Iwasawa

Akaljot Singh

Yuko Horio

Jorge O Munera

Takanori Takebe

Aaron M Zorn

Michael A Helmrath

Lee A Denson

James M Wells

Affiliations

Soon will be listed here.

Abstract

Background & Aims: Organs of the gastrointestinal tract contain tissue-resident immune cells that function during tissue development, homeostasis, and disease. However, most published human organoid model systems lack resident immune cells, thus limiting their potential as disease avatars. For example, human intestinal organoids (HIOs) derived from pluripotent stem cells contain epithelial and various mesenchymal cell types but lack immune cells. In this study, we aimed to develop an HIO model with functional tissue-resident macrophages.

Methods: HIOs and macrophages were generated separately through the directed differentiation of human pluripotent stem cells and combined in vitro. Following 2 weeks of coculture, the organoids were used for transcriptional profiling, functional analysis of macrophages, or transplanted into immunocompromised mice and matured in vivo for an additional 10-12 weeks.

Results: Macrophages were incorporated into developing HIOs and persisted for 2 weeks in vitro HIOs and for at least 12 weeks in HIOs in vivo. These cocultured macrophages had a transcriptional signature that resembled those in the human fetal intestine, indicating that they were acquiring the features of tissue-resident macrophages. HIO macrophages could phagocytose bacteria and produced inflammatory cytokines in response to proinflammatory signals, such as lipopolysaccharide, which could be reversed with interleukin-10.

Conclusions: We generated an HIO system containing functional tissue-resident macrophages for an extended period. This new organoid system can be used to investigate the molecular mechanisms involved in inflammatory bowel disease.

References

Arango Duque G, Descoteaux A . Macrophage cytokines: involvement in immunity and infectious diseases. Front Immunol. 2014; 5:491. PMC: 4188125. DOI: 10.3389/fimmu.2014.00491. View

Kakni P, Truckenmuller R, Habibovic P, van Griensven M, Giselbrecht S . A Microwell-Based Intestinal Organoid-Macrophage Co-Culture System to Study Intestinal Inflammation. Int J Mol Sci. 2022; 23(23). PMC: 9736416. DOI: 10.3390/ijms232315364. View

Yu Q, Kilik U, Holloway E, Tsai Y, Harmel C, Wu A . Charting human development using a multi-endodermal organ atlas and organoid models. Cell. 2021; 184(12):3281-3298.e22. PMC: 8208823. DOI: 10.1016/j.cell.2021.04.028. View

Tsai Y, Nattiv R, Dedhia P, Nagy M, Chin A, Thomson M . patterning of pluripotent stem cell-derived intestine recapitulates human development. Development. 2016; 144(6):1045-1055. PMC: 5358103. DOI: 10.1242/dev.138453. View

Gray J, Farber D . Tissue-Resident Immune Cells in Humans. Annu Rev Immunol. 2022; 40:195-220. PMC: 11194112. DOI: 10.1146/annurev-immunol-093019-112809. View

Gomez Perdiguero E, Klapproth K, Schulz C, Busch K, Azzoni E, Crozet L . Tissue-resident macrophages originate from yolk-sac-derived erythro-myeloid progenitors. Nature. 2014; 518(7540):547-51. PMC: 5997177. DOI: 10.1038/nature13989. View

Ruiz J, Chen G, Haro Mora J, Keyvanfar K, Liu C, Zou J . Robust generation of erythroid and multilineage hematopoietic progenitors from human iPSCs using a scalable monolayer culture system. Stem Cell Res. 2019; 41:101600. PMC: 6953424. DOI: 10.1016/j.scr.2019.101600. View

Dick S, Wong A, Hamidzada H, Nejat S, Nechanitzky R, Vohra S . Three tissue resident macrophage subsets coexist across organs with conserved origins and life cycles. Sci Immunol. 2022; 7(67):eabf7777. DOI: 10.1126/sciimmunol.abf7777. View

Tsuruta S, Kawasaki T, Machida M, Iwatsuki K, Inaba A, Shibata S . Development of Human Gut Organoids With Resident Tissue Macrophages as a Model of Intestinal Immune Responses. Cell Mol Gastroenterol Hepatol. 2022; 14(3):726-729.e5. PMC: 9421619. DOI: 10.1016/j.jcmgh.2022.06.006. View

10.

Workman M, Mahe M, Trisno S, Poling H, Watson C, Sundaram N . Engineered human pluripotent-stem-cell-derived intestinal tissues with a functional enteric nervous system. Nat Med. 2016; 23(1):49-59. PMC: 5562951. DOI: 10.1038/nm.4233. View

11.

Bain C, Bravo-Blas A, Scott C, Gomez Perdiguero E, Geissmann F, Henri S . Constant replenishment from circulating monocytes maintains the macrophage pool in the intestine of adult mice. Nat Immunol. 2014; 15(10):929-937. PMC: 4169290. DOI: 10.1038/ni.2967. View

12.

Davies L, Jenkins S, Allen J, Taylor P . Tissue-resident macrophages. Nat Immunol. 2013; 14(10):986-95. PMC: 4045180. DOI: 10.1038/ni.2705. View

13.

Bain C, Scott C, Uronen-Hansson H, Gudjonsson S, Jansson O, Grip O . Resident and pro-inflammatory macrophages in the colon represent alternative context-dependent fates of the same Ly6Chi monocyte precursors. Mucosal Immunol. 2012; 6(3):498-510. PMC: 3629381. DOI: 10.1038/mi.2012.89. View

14.

Eicher A, Kechele D, Sundaram N, Berns H, Poling H, Haines L . Functional human gastrointestinal organoids can be engineered from three primary germ layers derived separately from pluripotent stem cells. Cell Stem Cell. 2021; 29(1):36-51.e6. PMC: 8741755. DOI: 10.1016/j.stem.2021.10.010. View

15.

Varol C, Mildner A, Jung S . Macrophages: development and tissue specialization. Annu Rev Immunol. 2015; 33:643-75. DOI: 10.1146/annurev-immunol-032414-112220. View

16.

Garrett W, Gordon J, Glimcher L . Homeostasis and inflammation in the intestine. Cell. 2010; 140(6):859-70. PMC: 2845719. DOI: 10.1016/j.cell.2010.01.023. View

17.

Noel G, Baetz N, Staab J, Donowitz M, Kovbasnjuk O, Pasetti M . Erratum: A primary human macrophage-enteroid co-culture model to investigate mucosal gut physiology and host-pathogen interactions. Sci Rep. 2017; 7:46790. PMC: 5414479. DOI: 10.1038/srep46790. View

18.

Shaw T, Houston S, Wemyss K, Bridgeman H, Barbera T, Zangerle-Murray T . Tissue-resident macrophages in the intestine are long lived and defined by Tim-4 and CD4 expression. J Exp Med. 2018; 215(6):1507-1518. PMC: 5987925. DOI: 10.1084/jem.20180019. View

19.

Singh A, Poling H, Chaturvedi P, Thorner K, Sundaram N, Kechele D . Transplanted human intestinal organoids: a resource for modeling human intestinal development. Development. 2023; 150(9). PMC: 10259511. DOI: 10.1242/dev.201416. View

20.

Gritz E, Hirschi K . Specification and function of hemogenic endothelium during embryogenesis. Cell Mol Life Sci. 2016; 73(8):1547-67. PMC: 4805691. DOI: 10.1007/s00018-016-2134-0. View