Neurotensin stimulates sortilin and mTOR in human microglia inhibitable by methoxyluteolin, a potential therapeutic target for autism.
We had reported elevated serum levels of the peptide neurotensin (NT) in children with autism spectrum disorders (ASD). Here, we show that NT stimulates primary human microglia, the resident immune cells of the brain, and the immortalized cell line of human microglia-SV40. NT (10 nM) increases the gene expression and release (P< 0.001) of the proinflammatory cytokine IL-1β and chemokine (C-X-C motif) ligand 8 (CXCL8), chemokine (C-C motif) ligand 2 (CCL2), and CCL5 from human microglia. NT also stimulates proliferation (P< 0.05) of microglia-SV40. Microglia express only the receptor 3 (NTR3)/sortilin and not the NTR1 or NTR2. The use of siRNA to target sortilin reduces (P< 0.001) the NT-stimulated cytokine and chemokine gene expression and release from human microglia. Stimulation with NT (10 nM) increases the gene expression of sortilin (P< 0.0001) and causes the receptor to be translocated from the cytoplasm to the cell surface, and to be secreted extracellularly. Our findings also show increased levels of sortilin (P< 0.0001) in the serum from children with ASD (n= 36), compared with healthy controls (n= 20). NT stimulation of microglia-SV40 causes activation of the mammalian target of rapamycin (mTOR) signaling kinase, as shown by phosphorylation of its substrates and inhibition of these responses by drugs that prevent mTOR activation. NT-stimulated responses are inhibited by the flavonoid methoxyluteolin (0.1-1 μM). The data provide a link between sortilin and the pathological findings of microglia and inflammation of the brain in ASD. Thus, inhibition of this pathway using methoxyluteolin could provide an effective treatment of ASD.
Molecular Immunopharmacology and Drug Discovery Laboratory, Department of Integrative Physiology and Pathobiology, Tufts University School of Medicine, Boston, MA 02111.|Graduate Program in Cell, Molecular, and Developmental Biology, Sackler School of Graduate Biomedical Sciences, Tufts University, Boston, MA 02111.
Arti B
AB
Patel
Molecular Immunopharmacology and Drug Discovery Laboratory, Department of Integrative Physiology and Pathobiology, Tufts University School of Medicine, Boston, MA 02111.
Irene
I
Tsilioni
Department of Pharmacology, Boston University School of Medicine, Boston, MA 02118; theoharis.theoharides@tufts.edu sleeman@bu.edu.
Susan E
http://orcid.org/0000-0003-2875-1394
SE
Leeman
Molecular Immunopharmacology and Drug Discovery Laboratory, Department of Integrative Physiology and Pathobiology, Tufts University School of Medicine, Boston, MA 02111; theoharis.theoharides@tufts.edu sleeman@bu.edu.|Graduate Program in Cell, Molecular, and Developmental Biology, Sackler School of Graduate Biomedical Sciences, Tufts University, Boston, MA 02111.|Department of Internal Medicine, Tufts University School of Medicine and Tufts Medical Center, Boston, MA 02111.|Department of Psychiatry, Tufts University School of Medicine and Tufts Medical Center, Boston, MA 02111.
Theoharis C
TC
Theoharides
autism spectrum disorders; human microglia; mTOR; methoxyluteolin; sortilin
Nat Rev Neurosci. 2015 Aug;16(8):469-86
26189694
Immunity. 2008 Oct 17;29(4):565-77
18848473
Brain Behav Immun. 2011 Jan;25(1):40-5
20705131
FEBS Lett. 2001 Apr 20;495(1-2):100-5
11322955
Nat Neurosci. 2013 Nov;16(11):1537-43
24165680
Exp Cell Res. 2015 Oct 15;338(1):119-25
26004870
Exp Ther Med. 2014 May;7(5):1065-1070
24940388
Mol Autism. 2014 Jan 10;5(1):3
24410870
J Allergy Clin Immunol. 2015 Apr;135(4):1044-52.e5
25498791
Pharmacol Rev. 2000 Dec;52(4):673-751
11121513
J Neuroinflammation. 2010 Aug 23;7:48
20731814
Trends Pharmacol Sci. 1999 Jul;20(7):302-9
10390649
Mol Neurobiol. 2014 Jun;49(3):1487-500
24752587
J Neurosci. 2003 Feb 15;23(4):1198-205
12598608
Neuron Glia Biol. 2011 May;7(2-4):205-13
22874006
Biochem Biophys Res Commun. 2015 Oct 2;465(4):864-70
26319432
J Clin Psychopharmacol. 2008 Oct;28(5):479-83
18794641
J Neurosci. 2015 Oct 14;35(41):13836-42
26468183
Brain Res Mol Brain Res. 1998 Jun 1;57(1):1-9
9630473
Nat Rev Drug Discov. 2013 Oct;12(10):777-90
24080699
J Neurosci. 1996 Sep 15;16(18):5613-20
8795617
Science. 2011 Sep 9;333(6048):1456-8
21778362
Proc Natl Acad Sci U S A. 2013 Feb 26;110(9):3507-12
23401516
Transl Psychiatry. 2015 Sep 29;5:e647
26418275
Proc Natl Acad Sci U S A. 2012 Jul 31;109(31):12776-81
22802640
Nature. 1990 Jan 25;343(6256):336-40
2137200
J Neuroimmunol. 2009 Feb 15;207(1-2):111-6
19157572
Brain Behav Immun. 2013 Jul;31:54-68
22841693
J Clin Invest. 2011 Apr;121(4):1231-41
21490404
J Clin Invest. 1999 Mar;103(6):843-9
10079105
Genet Med. 2009 Feb;11(2):111-7
19265751
Annu Rev Clin Psychol. 2014;10:193-212
24329180
J Immunol. 2008 Apr 1;180(7):4586-95
18354181
Neuron. 2006 May 4;50(3):377-88
16675393
Transl Psychiatry. 2014 Oct 14;4:e466
25313509
Front Cell Neurosci. 2013 Mar 26;7:28
23533374
J Neuroimmunol. 2009 Dec 10;217(1-2):20-7
19766327
Science. 2016 Feb 26;351(6276):933-9
26822608
PLoS One. 2014 Oct 17;9(10):e110356
25330013
Br J Pharmacol. 1995 Apr;114(7):1466-70
7541694
J Biol Chem. 1973 Oct 10;248(19):6854-61
4745447
Biochem J. 2013 Mar 15;450(3):537-46
23252429
J Med Genet. 2005 Apr;42(4):318-21
15805158
Nat Commun. 2014 Dec 10;5:5748
25494366
Proc Natl Acad Sci U S A. 2006 May 16;103(20):7759-64
16682628
Biochem Soc Trans. 2015 Aug;43(4):702-6
26551716
Proc Natl Acad Sci U S A. 1987 May;84(10):3516-20
3472221
Biochem Pharmacol. 2009 Nov 1;78(9):1242-51
19576187
N Engl J Med. 2015 Jul 9;373(2):163-72
26154789
FASEB J. 2012 Aug;26(8):3103-17
22516295
Eur J Biochem. 1994 Apr 1;221(1):297-306
7909519
Glia. 2014 May;62(5):667-79
24504982
Genes Dev. 2008 Jun 1;22(11):1490-500
18519641
Transl Psychiatry. 2016 Jun 28;6(6):e844
27351598
Ann Neurol. 2012 Apr;71(4):444-57
22334391
J Neuroinflammation. 2010 Jan 14;7:3
20074346
Regul Pept. 1994 Aug 31;53(1):17-29
7800856
Proc Natl Acad Sci U S A. 2008 May 27;105(21):7534-9
18490655
Eur J Pharmacol. 2016 May 5;778:96-102
25941080
Lancet. 2014 Mar 8;383(9920):896-910
24074734
Trends Immunol. 2015 Jan;36(1):21-9
25592035
Curr Opin Neurobiol. 2015 Feb;30:92-9
25464374
Gut. 2008 Jan;57(1):50-8
17525093
Cell Rep. 2014 Jun 12;7(5):1626-39
24857653
Mol Pharm. 2007 Nov-Dec;4(6):826-32
17958394
Brain Res Mol Brain Res. 1999 Mar 5;65(2):216-9
10064893
Immunol Rev. 2012 Jul;248(1):228-39
22725965
J Neurosci Res. 2005 Aug 1;81(3):322-6
15957186
Trends Neurosci. 2014 Mar;37(3):125-35
24388427
J Neuroinflammation. 2013 Apr 09;10:46
23570274
Ann Neurol. 2005 Jan;57(1):67-81
15546155
Int J Immunopathol Pharmacol. 2012 Apr-Jun;25(2):317-23
22697063
J Neurosci Res. 2004 Oct 1;78(1):92-9
15372498
Clin Ther. 2013 May;35(5):592-602
23688534
Life Sci. 1990;46(9):607-17
2407920
Biochem Biophys Res Commun. 2002 Nov 15;298(5):760-4
12419319
Natl Health Stat Report. 2015 Nov 13;(87):1-20
26632847
J Clin Psychopharmacol. 2004 Dec;24(6):577-81
15538117
Biol Psychiatry. 2010 Aug 15;68(4):368-76
20674603
Immunology. 2014 Mar;141(3):314-27
24032675
Exp Neurobiol. 2015 Sep;24(3):177-85
26412966
J Biol Chem. 1997 Feb 7;272(6):3599-605
9013611
FEBS Lett. 1996 May 20;386(2-3):91-4
8647296
Mol Psychiatry. 2015 Apr;20(4):440-6
24934179
JAMA Psychiatry. 2013 Jan;70(1):49-58
23404112
J Pharmacol Exp Ther. 2002 Dec;303(3):1061-6
12438528
Cell Physiol Biochem. 2012;29(5-6):931-40
22613992
Trends Immunol. 2015 Oct;36(10):614-24
26431939
Mol Autism. 2016 Jan 20;7:9
26793298
J Nutr Biochem. 2011 Jul;22(7):612-24
21036586
Cell Metab. 2014 Mar 4;19(3):373-9
24508508
Pediatr Neurol. 2005 Sep;33(3):195-201
16139734
J Pharmacol Exp Ther. 2007 Mar;320(3):1237-45
17194798
Mol Psychiatry. 2002;7 Suppl 2:S12-3
12142934