N-Arachidonoyl Dopamine Modulates Acute Systemic Inflammation via Nonhematopoietic TRPV1 This information is current as Samira K. Lawton, Fengyun Xu, Alphonso Tran, Erika of October 1, 2021. Wong, Arun Prakash, Mark Schumacher, Judith Hellman and Kevin Wilhelmsen J Immunol 2017; 199:1465-1475; Prepublished online 12 July 2017; doi: 10.4049/jimmunol.1602151 http://www.jimmunol.org/content/199/4/1465 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2017/07/12/jimmunol.160215 Material 1.DCSupplemental http://www.jimmunol.org/ References This article cites 69 articles, 11 of which you can access for free at: http://www.jimmunol.org/content/199/4/1465.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on October 1, 2021 • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Author Choice Freely available online through The Journal of Immunology Author Choice option Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2017 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology N-Arachidonoyl Dopamine Modulates Acute Systemic Inflammation via Nonhematopoietic TRPV1 Samira K. Lawton,*,† Fengyun Xu,† Alphonso Tran,† Erika Wong,† Arun Prakash,† Mark Schumacher,† Judith Hellman,†,‡,1 and Kevin Wilhelmsen†,1 N-Arachidonoyl dopamine (NADA) is an endogenous lipid that potently activates the transient receptor potential vanilloid 1 (TRPV1), which mediates pain and thermosensation. NADA is also an agonist of cannabinoid receptors 1 and 2. We have reported that NADA reduces the activation of cultured human endothelial cells by LPS and TNF-a. Thus far, in vivo studies using NADA have focused on its neurologic and behavioral roles. In this article, we show that NADA potently decreases in vivo systemic inflammatory responses and levels of the coagulation intermediary plasminogen activator inhibitor 1 in three mouse models of inflammation: LPS, bacterial lipopeptide, and polymicrobial intra-abdominal sepsis. We also found that the administration of NADA increases survival in endotoxemic mice. Additionally, NADA reduces blood levels of the neuropeptide calcitonin gene- Downloaded from related peptide but increases the neuropeptide substance P in LPS-treated mice. We demonstrate that the anti-inflammatory effects of NADA are mediated by TRPV1 expressed by nonhematopoietic cells and provide data suggesting that neuronal TRPV1 may mediate NADA’s anti-inflammatory effects. These results indicate that NADA has novel TRPV1-dependent anti-inflammatory properties and suggest that the endovanilloid system might be targeted therapeutically in acute inflammation. The Journal of Immunology, 2017, 199: 1465–1475. he endogenous lipid N-arachidonoyl dopamine (NADA) (9–13). The peripheral terminals of TRPV1-expressing noci- http://www.jimmunol.org/ is composed of an arachidonic acid backbone conjugated ceptors innervate most organs and tissues. TRPV1 expressed in the T to a dopamine moiety (1). NADA is a putative endocanna- peripheral nervous system functions to integrate multiple noxious binoid and possesses activity via the G protein–coupled cannabinoid stimuli, ultimately leading to the release of neuropeptides, such as receptors (2, 3). NADA has also been classified as an endovanilloid, calcitonin gene–related peptide (CGRP) and substance P. These which is a group of endogenous activators of the transient receptor neuropeptides elicit pain and neurogenic inflammatory responses potential vanilloid 1 (TRPV1) and include the endocannabinoid (14–18). TRPV1 is also expressed in the CNS, including the spinal anandamide (AEA) and a variety of lipoxygenase products (4–6). cord, striatum, hippocampus, cerebellum, and amygdala (19–23). TRPV1, a nonselective cation channel, is primarily known for its role Finally, TRPV1 is found in nonneuronal cells, including leukocytes, by guest on October 1, 2021 in sensing pain and temperature. Of the known endovanilloids, smooth muscle cells, and endothelial cells (11, 17, 24–27). In this NADA is the most potent activator of TRPV1 and is considered to be regard, TRPV1 activation has been reported to regulate the acti- a principal endogenous TRPV1 ligand (1, 5, 7, 8). TRPV1 is also vation and proinflammatory properties of CD4+ T cells (28) activated by a variety of noxious insults, including heat . 42˚C, NADA and TRPV1 have overlapping distributions, because low pH, and capsaicin, the active ingredient in chili peppers (5). NADA has been detected in the striatum, cerebellum, hippocam- TRPV1 is highly expressed in the peripheral nervous system, pus, thalamus, brainstem, and dorsal root ganglia (7, 19, 20, 23, specifically in a subset of small-diameter primary sensory neurons 29). Moreover, NADA has been proposed to modulate neuronal in trigeminal nerve and dorsal root ganglia that detect noxious homeostasis by reducing or inducing cation influx via activation of stimuli and in the inferior (nodose) ganglion of the vagus nerve cannabinoid receptors and TRPV1, respectively (1, 30). In addition *Biomedical Sciences Graduate Program, University of California, San Francisco, TLR agonist challenge experiments; E.W. and F.X. performed cecal ligation and San Francisco, CA 94143; †Department of Anesthesia and Perioperative Care, Uni- puncture surgeries; A.T., S.K.L., and E.W. collected and processed plasma samples versity of California, San Francisco, San Francisco, CA 94143; and ‡Division of and performed ELISAs; K.W., A.P., and S.K.L. performed the multiplex experiments; Critical Care Medicine, University of California, San Francisco, San Francisco, and K.W. and S.K.L. performed the ex vivo experiments. M.S. provided input into the CA 94143 design of studies and contributed to the writing of the manuscript. All authors pro- vided input on experimental design. 1J.H. and K.W. are cosenior authors. Address correspondence and reprint requests to Dr. Judith Hellman, Department of ORCID: 0000-0002-1424-0932 (A.P.). Anesthesia, University of California, San Francisco, 500 Parnassus Avenue, Box Received for publication December 22, 2016. Accepted for publication June 13, 0648, San Francisco, CA 94143. E-mail address: [email protected] 2017. The online version of this article contains supplemental material. This work was supported by grants from the Department of Anesthesia and Perioper- Abbreviations used in this article: AEA, anandamide; 2-AG, 2-arachidonoyl glycerol; ative Care, University of California, San Francisco (to J.H.), the International Anes- CB, cannabinoid; CBR, CB receptor; CB R, CBR 1; CB R, CBR 2; CD45.1+, B6. thesia Research Society (to J.H.), National Science Foundation Graduate Research 1 2 SJL-Ptprca mice; CGRP, calcitonin gene–related peptide; CLP, cecal ligation and Fellowship Program Grant 1144247 (to S.K.L.), and the University of California, San puncture; CV, coefficient of variability; FCSB, Flow Cytometry Staining Buffer; Francisco Research Evaluation and Allocation Committee/Research Allocation Pro- LC-MS/MS, liquid chromatography–tandem mass spectrometry; NADA, N-arachi- gram (Huntington Fund; to J.H.). donoyl dopamine; PAI, plasminogen activator inhibitor; TRPV1, transient receptor K.W. and J.H. conceptualized the project; K.W. conceptualized the design of exper- potential vanilloid 1; Trpv12/2, B6.129 3 1-Trpv1tm1Jul/J; WT, wild-type. iments, analyzed data, and contributed to the writing of the manuscript; J.H. super- This article is distributed under The American Association of Immunologists, Inc., vised the project, acquired funding, and analyzed data; S.K.L. designed and Reuse Terms and Conditions for Author Choice articles. performed experiments, including the neuropeptide enzyme immunoassay experi- ments, generated bone marrow chimera mice, performed flow cytometry, and ana- Ó lyzed data. S.K.L. and J.H. wrote the manuscript. S.K.L., F.X., and A.T. conducted Copyright 2017 by The American Association of Immunologists, Inc. 0022-1767/17/$30.00 www.jimmunol.org/cgi/doi/10.4049/jimmunol.1602151 1466 NADA REDUCES INFLAMMATION IN SEPSIS VIA NONMYELOID TRPV1 to its neurologic and behavioral effects, limited studies suggest that Immunoassays and MTT assay NADA has anti-inflammatory effects on nonneuronal cells, such as IL-6, CCL-2, IL-10, and plasminogen activator inhibitor (PAI)-1 levels were astrocytes, leukocytes, and endothelial cells (2, 24, 31–34). quantified in mouse plasma samples by ELISA (R&D Systems, Minne- We previously found that treatment with NADA reduces the acti- apolis, MN and Innovative Research, Novi, MI). Substance P and CGRPa/b vation of cultured endothelial cells by TNF-a, as well as by bacterial levels were quantified in mouse plasma samples by enzyme immunoassay lipopeptides (TLR2 agonists) and LPS (endotoxin, TLR4 agonist) (Enzo Life Sciences, Farmingdale, NY and Cayman Chemicals, Ann Arbor, MI, respectively), and absorbance was read using a FLUOstar OPTIMA (24). Furthermore, our data using pharmacological