TY - JOUR
T1 - Identification of Natural RORγ Ligands that Regulate the Development of Lymphoid Cells
AU - Santori, Fabio R.
AU - Huang, Pengxiang
AU - Van De Pavert, Serge A.
AU - Douglass, Eugene F.
AU - Leaver, David J.
AU - Haubrich, Brad A.
AU - Keber, Rok
AU - Lorbek, Gregor
AU - Konijn, Tanja
AU - Rosales, Brittany N.
AU - Rozman, Damjana
AU - Horvat, Simon
AU - Rahier, Alain
AU - Mebius, Reina E.
AU - Rastinejad, Fraydoon
AU - Nes, W. David
AU - Littman, Dan R.
N1 - Funding Information:
We thank David Mangelsdorf for advice; The Scripps center for mass spectrometry and metabolomics for LC-MS and XCMS analysis; Gesine Saher for the sxlt cell line; Jun Huh for plasmids; Dr. Rene Lafont for ecdysone biosynthetic intermediates; Jialin Liu for chemical synthesis; Raghu R.V. Malapaka and Eric H. Xu for help with alphascreen assays; Jason A. Hall for mouse breeding and genotyping; Wendy Huang for experimental support; and Renee de Pooter, Maria Ciofani, and Natalia B. Ivanova for critical reading of the manuscript. This work was supported by NIH grant R01-AI080885-01A1 (D.R.L.), National Science Foundation grant MCB-0929212 (W.D.N), Slovenian Research Agency (ARRS) Grants J3-6804 (S.H,), J4-4306 (S.H, and R.K.), and P1-0104 (D.R.) and graduate fellowship (G.L.), breakthrough project NGI 40-41009-98-9077 (S.A.v.d.P.), Netherlands Organization for Scientific Research-Earth and Life Sciences (NWO-ALW) Top Grant 09.048 (R.E.M.), and NIH grant T32HL007151 (F.R.S.). D.R.L. is an Investigator of the Howard Hughes Medical Institute.
Publisher Copyright:
© 2015 Elsevier Inc.
PY - 2015/2/3
Y1 - 2015/2/3
N2 - Summary 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.
AB - Summary 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.
UR - http://www.scopus.com/inward/record.url?scp=84922901154&partnerID=8YFLogxK
U2 - 10.1016/j.cmet.2015.01.004
DO - 10.1016/j.cmet.2015.01.004
M3 - Article
C2 - 25651181
AN - SCOPUS:84922901154
VL - 21
SP - 286
EP - 298
JO - Cell Metabolism
JF - Cell Metabolism
SN - 1550-4131
IS - 2
ER -