Identification of Natural RORγ Ligands That Regulate the Development of Lymphoid Cells

Overview

Journal Cell Metab

Publisher Cell Press

Specialties Cell Biology
Endocrinology

Date 2015 Feb 5

PMID 25651181

Citations 124

Authors

Fabio R Santori

Pengxiang Huang

Serge A van de Pavert

Eugene F Douglass Jr

David J Leaver

Brad A Haubrich

Rok Keber

Gregor Lorbek

Tanja Konijn

Brittany N Rosales

Damjana Rozman

Simon Horvat

Alain Rahier

Reina E Mebius

Fraydoon Rastinejad

W David Nes

Dan R Littman

Affiliations

Soon will be listed here.

Abstract

Mice deficient in the nuclear hormone receptor RORγt have defective development of thymocytes, lymphoid organs, Th17 cells, and type 3 innate lymphoid cells. RORγt binds to oxysterols derived from cholesterol catabolism, but it is not clear whether these are its natural ligands. Here, we show that sterol lipids are necessary and sufficient to drive RORγt-dependent transcription. We combined overexpression, RNAi, and genetic deletion of metabolic enzymes to study RORγ-dependent transcription. Our results are consistent with the RORγt ligand(s) being a cholesterol biosynthetic intermediate (CBI) downstream of lanosterol and upstream of zymosterol. Analysis of lipids bound to RORγ identified molecules with molecular weights consistent with CBIs. Furthermore, CBIs stabilized the RORγ ligand-binding domain and induced coactivator recruitment. Genetic deletion of metabolic enzymes upstream of the RORγt-ligand(s) affected the development of lymph nodes and Th17 cells. Our data suggest that CBIs play a role in lymphocyte development potentially through regulation of RORγt.

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References

Huang P, Chandra V, Rastinejad F . Structural overview of the nuclear receptor superfamily: insights into physiology and therapeutics. Annu Rev Physiol. 2010; 72:247-72. PMC: 3677810. DOI: 10.1146/annurev-physiol-021909-135917. View

Eberl G, Littman D . Thymic origin of intestinal alphabeta T cells revealed by fate mapping of RORgammat+ cells. Science. 2004; 305(5681):248-51. DOI: 10.1126/science.1096472. View

Bloch K . The biological synthesis of cholesterol. Science. 1965; 150(3692):19-28. DOI: 10.1126/science.150.3692.19. View

Cooke J, Sang J . Utilization of sterols by larvae of Drosophila melanogaster. J Insect Physiol. 1970; 16(5):801-12. DOI: 10.1016/0022-1910(70)90214-3. View

WOSTMANN B, PLEASANTS J, Bealmear P, Kincade P . Serum proteins and lymphoid tissues in germ-free mice fed a chemically defined, water soluble, low molecular weight diet. Immunology. 1970; 19(3):443-8. PMC: 1455676. View

Silberkang M, Havel C, Friend D, McCarthy B, Watson J . Isoprene synthesis in isolated embryonic Drosophila cells. I. Sterol-deficient eukaryotic cells. J Biol Chem. 1983; 258(13):8503-11. View

Billheimer J, Chamoun D, Esfahani M . Defective 3-ketosteroid reductase activity in a human monocyte-like cell line. J Lipid Res. 1987; 28(6):704-9. View

Goldstein J, Brown M . Regulation of the mevalonate pathway. Nature. 1990; 343(6257):425-30. DOI: 10.1038/343425a0. View

Koelle M, Talbot W, Segraves W, Bender M, Cherbas P, Hogness D . The Drosophila EcR gene encodes an ecdysone receptor, a new member of the steroid receptor superfamily. Cell. 1991; 67(1):59-77. DOI: 10.1016/0092-8674(91)90572-g. View

10.

Ivanov I, McKenzie B, Zhou L, Tadokoro C, Lepelley A, Lafaille J . The orphan nuclear receptor RORgammat directs the differentiation program of proinflammatory IL-17+ T helper cells. Cell. 2006; 126(6):1121-33. DOI: 10.1016/j.cell.2006.07.035. View

11.

Wassif C, Brownson K, Sterner A, Forlino A, Zerfas P, Wilson W . HEM dysplasia and ichthyosis are likely laminopathies and not due to 3beta-hydroxysterol Delta14-reductase deficiency. Hum Mol Genet. 2007; 16(10):1176-87. DOI: 10.1093/hmg/ddm065. View

12.

Saher G, Quintes S, Mobius W, Wehr M, Kramer-Albers E, Brugger B . Cholesterol regulates the endoplasmic reticulum exit of the major membrane protein P0 required for peripheral myelin compaction. J Neurosci. 2009; 29(19):6094-104. PMC: 6665514. DOI: 10.1523/JNEUROSCI.0686-09.2009. View

13.

Bauman D, Bitmansour A, McDonald J, Thompson B, Liang G, Russell D . 25-Hydroxycholesterol secreted by macrophages in response to Toll-like receptor activation suppresses immunoglobulin A production. Proc Natl Acad Sci U S A. 2009; 106(39):16764-9. PMC: 2757821. DOI: 10.1073/pnas.0909142106. View

14.

van de Pavert S, Olivier B, Goverse G, Vondenhoff M, Greuter M, Beke P . Chemokine CXCL13 is essential for lymph node initiation and is induced by retinoic acid and neuronal stimulation. Nat Immunol. 2009; 10(11):1193-9. PMC: 2771164. DOI: 10.1038/ni.1789. View

15.

Wang Y, Kumar N, Solt L, Richardson T, Helvering L, Crumbley C . Modulation of retinoic acid receptor-related orphan receptor alpha and gamma activity by 7-oxygenated sterol ligands. J Biol Chem. 2009; 285(7):5013-25. PMC: 2836105. DOI: 10.1074/jbc.M109.080614. View

16.

Wang Y, Kumar N, Crumbley C, Griffin P, Burris T . A second class of nuclear receptors for oxysterols: Regulation of RORalpha and RORgamma activity by 24S-hydroxycholesterol (cerebrosterol). Biochim Biophys Acta. 2010; 1801(8):917-23. PMC: 2886165. DOI: 10.1016/j.bbalip.2010.02.012. View

17.

He M, Kratz L, Michel J, Vallejo A, Ferris L, Kelley R . Mutations in the human SC4MOL gene encoding a methyl sterol oxidase cause psoriasiform dermatitis, microcephaly, and developmental delay. J Clin Invest. 2011; 121(3):976-84. PMC: 3049385. DOI: 10.1172/JCI42650. View

18.

Huh J, Leung M, Huang P, Ryan D, Krout M, Malapaka R . Digoxin and its derivatives suppress TH17 cell differentiation by antagonizing RORγt activity. Nature. 2011; 472(7344):486-90. PMC: 3172133. DOI: 10.1038/nature09978. View

19.

Skarnes W, Rosen B, West A, Koutsourakis M, Bushell W, Iyer V . A conditional knockout resource for the genome-wide study of mouse gene function. Nature. 2011; 474(7351):337-42. PMC: 3572410. DOI: 10.1038/nature10163. View

20.

Keber R, Motaln H, Wagner K, Debeljak N, Rassoulzadegan M, Acimovic J . Mouse knockout of the cholesterogenic cytochrome P450 lanosterol 14alpha-demethylase (Cyp51) resembles Antley-Bixler syndrome. J Biol Chem. 2011; 286(33):29086-29097. PMC: 3190716. DOI: 10.1074/jbc.M111.253245. View