Advertisement

Longitudinal Transcriptomic Profiling in Carrageenan-Induced Rat Hind Paw Peripheral Inflammation and Hyperalgesia Reveals Progressive Recruitment of Innate Immune System Components

Published:November 20, 2020DOI:https://doi.org/10.1016/j.jpain.2020.11.001

      Highlights

      • Temporal gene regulatory programs with emphasis on the “inflammatory secretome.”
      • Sequential waves of leukocytes and antecedent patterns of the chemoattractants.
      • Subcellular localization of differentially expressed genes providing new targets.
      • Unique expression patterns of collagen-encoding genes related to tissue remodeling.

      Abstract

      Pain is a common but potentially debilitating symptom, often requiring complex management strategies. To understand the molecular dynamics of peripheral inflammation and nociceptive pain, we investigated longitudinal changes in behavior, tissue structure, and transcriptomic profiles in the rat carrageenan-induced peripheral inflammation model. Sequential changes in the number of differentially expressed genes are consistent with temporal recruitment of key leukocyte populations, mainly neutrophils and macrophages with each wave being preceded by upregulation of the cell-specific chemoattractants, Cxcl1 and Cxcl2, and Ccl2 and Ccl7, respectively. We defined 12 temporal gene clusters based on expression pattern. Within the patterns we extracted genes comprising the inflammatory secretome and others related to nociceptive tissue remodeling and to sensory perception of pain. Structural tissue changes, involving upregulation of multiple collagens occurred as soon as 1-hour postinjection, consistent with inflammatory tissue remodeling. Inflammatory expression profiling revealed a broad-spectrum, temporally orchestrated molecular and cellular recruitment process. The results provide numerous potential targets for modulation of pain and inflammation.

      Perspective

      This study investigates the highly orchestrated biological response during tissue inflammation with precise assessment of molecular dynamics at the transcriptional level. The results identify transcriptional changes that define an evolving inflammatory state in rats. This study provides foundational data for identifying markers of, and potential treatments for, inflammation and pain in patients.

      Key words

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to The Journal of Pain
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Bardina SV
        • Michlmayr D
        • Hoffman KW
        • Obara CJ
        • Sum J
        • Charo IF
        • Lu W
        • Pletnev AG
        • Lim JK
        Differential roles of chemokines CCL2 and CCL7 in monocytosis and leukocyte migration during west Nile virus infection.
        J Immunol. 2015; 195: 4306-4318
        • Barr TP
        • Hrnjic A
        • Khodorova A
        • Sprague JM
        • Strichartz GR
        Sensitization of cutaneous neuronal purinergic receptors contributes to endothelin-1-induced mechanical hypersensitivity.
        Pain. 2014; 155: 1091-1101
        • Barr TP
        • Kam S
        • Khodorova A
        • Montmayeur JP
        • Strichartz GR
        New perspectives on the endothelin axis in pain.
        Pharmacol Res. 2011; 63: 532-540
        • Binder JX
        • Pletscher-Frankild S
        • Tsafou K
        • Stolte C
        • O'Donoghue SI
        • Schneider R
        • Jensen LJ
        COMPARTMENTS: Unification and visualization of protein subcellular localization evidence.
        Database (Oxford). 2014; 2014: bau012
        • Blivis D
        • Haspel G
        • Mannes PZ
        • O'Donovan MJ
        • Iadarola MJ
        Identification of a novel spinal nociceptive-motor gate control for Adelta pain stimuli in rats.
        Elife. 2017; 6 (e23584)
        • Bruno K
        • Woller SA
        • Miller YI
        • Yaksh TL
        • Wallace M
        • Beaton G
        • Chakravarthy K
        Targeting toll-like receptor-4 (TLR4)-an emerging therapeutic target for persistent pain states.
        Pain. 2018; 159: 1908-1915
        • Carvalho TT
        • Borghi SM
        • Pinho-Ribeiro FA
        • Mizokami SS
        • Cunha TM
        • Ferreira SH
        • Cunha FQ
        • Casagrande R
        • Verri Jr, WA
        Granulocyte-colony stimulating factor (G-CSF)-induced mechanical hyperalgesia in mice: Role for peripheral TNFalpha, IL-1beta and IL-10.
        Eur J Pharmacol. 2015; 749: 62-72
        • Chazaud B
        Inflammation during skeletal muscle regeneration and tissue remodeling: Application to exercise-induced muscle damage management.
        Immunol Cell Biol. 2016; 94: 140-145
        • Cox MA
        • Jenh CH
        • Gonsiorek W
        • Fine J
        • Narula SK
        • Zavodny PJ
        • Hipkin RW
        Human interferon-inducible 10-kDa protein and human interferon-inducible T cell alpha chemoattractant are allotopic ligands for human CXCR3: Differential binding to receptor states.
        Mol Pharmacol. 2001; 59: 707-715
        • De Filippo K
        • Dudeck A
        • Hasenberg M
        • Nye E
        • van Rooijen N
        • Hartmann K
        • Gunzer M
        • Roers A
        • Hogg N
        Mast cell and macrophage chemokines CXCL1/CXCL2 control the early stage of neutrophil recruitment during tissue inflammation.
        Blood. 2013; 121: 4930-4937
        • de la Vega Gallardo N
        • Dittmer M
        • Dombrowski Y
        • Fitzgerald DC
        Regenerating CNS myelin: Emerging roles of regulatory T cells and CCN proteins.
        Neurochem Int. 2019; 130104349
        • Deftu AT
        • Ciorescu R
        • Gheorghe RO
        • Mihailescu D
        • Ristoiu V
        CXCL1 and CXCL2 inhibit the axon outgrowth in a time- and cell-type-dependent manner in adult rat dorsal root ganglia neurons.
        Neurochem Res. 2019; 44: 2215-2229
        • Gaskin DJ
        • Richard P
        The economic costs of pain in the United States.
        J Pain. 2012; 13: 715-724
        • Giannini E
        • Lattanzi R
        • Nicotra A
        • Campese AF
        • Grazioli P
        • Screpanti I
        • Balboni G
        • Salvadori S
        • Sacerdote P
        • Negri L
        The chemokine Bv8/prokineticin 2 is up-regulated in inflammatory granulocytes and modulates inflammatory pain.
        Proc Natl Acad Sci U S A. 2009; 106: 14646-14651
        • Gokin AP
        • Fareed MU
        • Pan HL
        • Hans G
        • Strichartz GR
        • Davar G
        Local injection of endothelin-1 produces pain-like behavior and excitation of nociceptors in rats.
        J Neurosci. 2001; 21: 5358-5366
        • Goswami SC
        • Mishra SK
        • Maric D
        • Kaszas K
        • Gonnella GL
        • Clokie SJ
        • Kominsky HD
        • Gross JR
        • Keller JM
        • Mannes AJ
        • Hoon MA
        • Iadarola MJ
        Molecular signatures of mouse TRPV1-lineage neurons revealed by RNA-Seq transcriptome analysis.
        J Pain. 2014; 15: 1338-1359
        • Gubbels Bupp MR
        Sex, the aging immune system, and chronic disease.
        Cell Immunol. 2015; 294: 102-110
        • Hoffmann T
        • Negri L
        • Maftei D
        • Lattanzi R
        • Reeh PW
        The prokineticin Bv8 sensitizes cutaneous terminals of female mice to heat.
        Eur J Pain. 2016; 20: 1326-1334
        • Holness CL
        • Simmons DL
        Molecular cloning of CD68, a human macrophage marker related to lysosomal glycoproteins.
        Blood. 1993; 81: 1607-1613
        • Inoue K
        • Hosoi J
        • Denda M
        Extracellular ATP has stimulatory effects on the expression and release of IL-6 via purinergic receptors in normal human epidermal keratinocytes.
        J Invest Dermatol. 2007; 127: 362-371
        • Khodorova A
        • Fareed MU
        • Gokin A
        • Strichartz GR
        • Davar G
        Local injection of a selective endothelin-B receptor agonist inhibits endothelin-1-induced pain-like behavior and excitation of nociceptors in a naloxone-sensitive manner.
        J Neurosci. 2002; 22: 7788-7796
        • Khodorova A
        • Montmayeur JP
        • Strichartz G
        Endothelin receptors and pain.
        J Pain. 2009; 10: 4-28
        • Koizumi S
        • Fujishita K
        • Inoue K
        • Shigemoto-Mogami Y
        • Tsuda M
        • Inoue K
        Ca2+ waves in keratinocytes are transmitted to sensory neurons: The involvement of extracellular ATP and P2Y2 receptor activation.
        Biochem J. 2004; 380: 329-338
        • Kroin JS
        • Buvanendran A
        • Nagalla SK
        • Tuman KJ
        Postoperative pain and analgesic responses are similar in male and female Sprague-Dawley rats.
        Can J Anaesth. 2003; 50: 904-908
        • Kubota S
        • Takigawa M
        Cellular and molecular actions of CCN2/CTGF and its role under physiological and pathological conditions.
        Clin Sci (Lond). 2015; 128: 181-196
        • LaPaglia DM
        • Sapio MR
        • Burbelo PD
        • Thierry-Mieg J
        • Thierry-Mieg D
        • Raithel SJ
        • Ramsden CE
        • Iadarola MJ
        • Mannes AJ
        RNA-Seq investigations of human post-mortem trigeminal ganglia.
        Cephalalgia. 2018; 38: 912-932
        • Lattanzi R
        • Maftei D
        • Marconi V
        • Florenzano F
        • Franchi S
        • Borsani E
        • Rodella LF
        • Balboni G
        • Salvadori S
        • Sacerdote P
        • Negri L
        Prokineticin 2 upregulation in the peripheral nervous system has a major role in triggering and maintaining neuropathic pain in the chronic constriction injury model.
        Biomed Res Int. 2015; 2015301292
        • Lima AL
        • Karl I
        • Giner T
        • Poppe H
        • Schmidt M
        • Presser D
        • Goebeler M
        • Bauer B
        Keratinocytes and neutrophils are important sources of proinflammatory molecules in hidradenitis suppurativa.
        Br J Dermatol. 2016; 174: 514-521
        • Liu S
        • Leask A
        CCN2 modulates hair follicle cycling in mice.
        Mol Biol Cell. 2013; 24: 3939-3944
        • Llorian-Salvador M
        • Gonzalez-Rodriguez S
        • Lastra A
        • Fernandez-Garcia MT
        • Hidalgo A
        • Menendez L
        • Baamonde A
        Involvement of CC chemokine receptor 1 and CCL3 in acute and chronic inflammatory pain in mice.
        Basic Clin Pharmacol Toxicol. 2016; 119: 32-40
        • Loppnow H
        • Libby P
        Adult human vascular endothelial cells express the IL6 gene differentially in response to LPS or IL1.
        Cell Immunol. 1989; 122: 493-503
        • Luo P
        • Shao J
        • Jiao Y
        • Yu W
        • Rong W
        CC chemokine ligand 2 (CCL2) enhances TTX-sensitive sodium channel activity of primary afferent neurons in the complete Freud adjuvant-induced inflammatory pain model.
        Acta Biochim Biophys Sin (Shanghai). 2018; 50: 1219-1226
        • Malin SA
        • Davis BM
        • Koerber HR
        • Reynolds IJ
        • Albers KM
        • Molliver DC
        Thermal nociception and TRPV1 function are attenuated in mice lacking the nucleotide receptor P2Y2.
        Pain. 2008; 138: 484-496
        • Moehring F
        • Cowie AM
        • Menzel AD
        • Weyer AD
        • Grzybowski M
        • Arzua T
        • Geurts AM
        • Palygin O
        • Stucky CL
        Keratinocytes mediate innocuous and noxious touch via ATP-P2X4 signaling.
        Elife. 2018; 7 (e31684)
        • Moehring F
        • Halder P
        • Seal RP
        • Stucky CL
        Uncovering the cells and circuits of touch in normal and pathological settings.
        Neuron. 2018; 100: 349-360
        • Morris CJ
        Carrageenan-induced paw edema in the rat and mouse.
        Methods Mol Biol. 2003; 225: 115-121
        • Nirmal AJ
        • Regan T
        • Shih BB
        • Hume DA
        • Sims AH
        • Freeman TC
        Immune cell gene signatures for profiling the microenvironment of solid tumors.
        Cancer Immunol Res. 2018; 6: 1388-1400
        • Peters NC
        • Egen JG
        • Secundino N
        • Debrabant A
        • Kimblin N
        • Kamhawi S
        • Lawyer P
        • Fay MP
        • Germain RN
        • Sacks D
        In vivo imaging reveals an essential role for neutrophils in leishmaniasis transmitted by sand flies.
        Science. 2008; 321: 970-974
        • Qi S
        • Zhao F
        • Li Z
        • Liang F
        • Yu S
        Silencing of PTX3 alleviates LPS-induced inflammatory pain by regulating TLR4/NF-kappaB signaling pathway in mice.
        Biosci Rep. 2020; 40 (BSR20194208)
        • Raghu H
        • Lepus CM
        • Wang Q
        • Wong HH
        • Lingampalli N
        • Oliviero F
        • Punzi L
        • Giori NJ
        • Goodman SB
        • Chu CR
        • Sokolove JB
        • Robinson WH
        CCL2/CCR2, but not CCL5/CCR5, mediates monocyte recruitment, inflammation and cartilage destruction in osteoarthritis.
        Ann Rheum Dis. 2017; 76: 914-922
        • Raithel SJ
        • Sapio MR
        • LaPaglia DM
        • Iadarola MJ
        • Mannes AJ
        Transcriptional changes in dorsal spinal cord persist after surgical incision despite preemptive analgesia with peripheral resiniferatoxin.
        Anesthesiology. 2018; 128: 620-635
        • Rastogi S
        • Rost B
        LocDB: experimental annotations of localization for Homo sapiens and Arabidopsis thaliana.
        Nucleic Acids Res. 2011; 39: D230-D234
        • Rose-John S
        Interleukin-6 family cytokines.
        Cold Spring Harb Perspect Biol. 2018; 10 (a028415)
        • Rosen S
        • Ham B
        • Mogil JS
        Sex differences in neuroimmunity and pain.
        J Neurosci Res. 2017; 95: 500-508
        • Sadler KE
        • Moehring F
        • Stucky CL
        Keratinocytes contribute to normal cold and heat sensation.
        Elife. 2020; 9 (e58625)
        • Sapio MR
        • Goswami SC
        • Gross JR
        • Mannes AJ
        • Iadarola MJ
        Transcriptomic analyses of genes and tissues in inherited sensory neuropathies.
        Exp Neurol. 2016; 283: 375-395
        • Sapio MR
        • Iadarola MJ
        • Loydpierson AJ
        • Kim JJ
        • Thierry-Mieg D
        • Thierry-Mieg J
        • Maric D
        • Mannes AJ
        Dynorphin and enkephalin opioid peptides and transcripts in spinal cord and dorsal root ganglion during peripheral inflammatory hyperalgesia and allodynia.
        J Pain. 2020; 21: 988-1004
        • Schweizerhof M
        • Stosser S
        • Kurejova M
        • Njoo C
        • Gangadharan V
        • Agarwal N
        • Schmelz M
        • Bali KK
        • Michalski CW
        • Brugger S
        • Dickenson A
        • Simone DA
        • Kuner R
        Hematopoietic colony-stimulating factors mediate tumor-nerve interactions and bone cancer pain.
        Nat Med. 2009; 15: 802-807
        • Silva RL
        • Lopes AH
        • Guimaraes RM
        • Cunha TM
        CXCL1/CXCR2 signaling in pathological pain: Role in peripheral and central sensitization.
        Neurobiol Dis. 2017; 105: 109-116
        • Smith JR
        • Hayman GT
        • Wang SJ
        • Laulederkind SJF
        • Hoffman MJ
        • Kaldunski ML
        • Tutaj M
        • Thota J
        • Nalabolu HS
        • Ellanki SLR
        • Tutaj MA
        • De Pons JL
        • Kwitek AE
        • Dwinell MR
        • Shimoyama ME
        The year of the rat: The rat genome database at 20: A multi-species knowledgebase and analysis platform.
        Nucleic Acids Res. 2020; 48: D731-D742
        • Tajerian M
        • Clark JD
        Spinal matrix metalloproteinase 8 regulates pain after peripheral trauma.
        J Pain Res. 2019; 12: 1133-1138
        • Talagas M
        • Lebonvallet N
        • Berthod F
        • Misery L
        Lifting the veil on the keratinocyte contribution to cutaneous nociception.
        Protein Cell. 2020; 11: 239-250
        • Thul PJ
        • Akesson L
        • Wiking M
        • Mahdessian D
        • Geladaki A
        • Ait Blal H
        • Alm T
        • Asplund A
        • Bjork L
        • Breckels LM
        • Backstrom A
        • Danielsson F
        • Fagerberg L
        • Fall J
        • Gatto L
        • Gnann C
        • Hober S
        • Hjelmare M
        • Johansson F
        • Lee S
        • Lindskog C
        • Mulder J
        • Mulvey CM
        • Nilsson P
        • Oksvold P
        • Rockberg J
        • Schutten R
        • Schwenk JM
        • Sivertsson A
        • Sjostedt E
        • Skogs M
        • Stadler C
        • Sullivan DP
        • Tegel H
        • Winsnes C
        • Zhang C
        • Zwahlen M
        • Mardinoglu A
        • Ponten F
        • von Feilitzen K
        • Lilley KS
        • Uhlen M
        • Lundberg E
        A subcellular map of the human proteome.
        Science. 2017; 356 (eaal3321)
        • Tsuda M
        • Kuboyama K
        • Inoue T
        • Nagata K
        • Tozaki-Saitoh H
        • Inoue K
        Behavioral phenotypes of mice lacking purinergic P2X4 receptors in acute and chronic pain assays.
        Mol Pain. 2009; 5: 28
        • UniProt Consortium T
        UniProt: The universal protein knowledgebase.
        Nucleic Acids Res. 2018; 46: 2699
        • Velly AM
        • Mohit S
        Epidemiology of pain and relation to psychiatric disorders.
        Prog Neuropsychopharmacol Biol Psychiatry. 2018; 87: 159-167
        • Wang S
        • Song R
        • Wang Z
        • Jing Z
        • Wang S
        • Ma J
        S100A8/A9 in inflammation.
        Front Immunol. 2018; 9: 1298
        • Yang F
        • Sun W
        • Luo WJ
        • Yang Y
        • Yang F
        • Wang XL
        • Chen J
        SDF1-CXCR4 signaling contributes to the transition from acute to chronic pain state.
        Mol Neurobiol. 2017; 54: 2763-2775
        • Yashima S
        • Shimazaki A
        • Mitoma J
        • Nakagawa T
        • Abe M
        • Yamada H
        • Higashi H
        Close association of B2 bradykinin receptors with P2Y2 ATP receptors.
        J Biochem. 2015; 158: 155-163
        • Zenz R
        • Eferl R
        • Kenner L
        • Florin L
        • Hummerich L
        • Mehic D
        • Scheuch H
        • Angel P
        • Tschachler E
        • Wagner EF
        Psoriasis-like skin disease and arthritis caused by inducible epidermal deletion of Jun proteins.
        Nature. 2005; 437: 369-375
        • Zhang N
        • Inan S
        • Cowan A
        • Sun R
        • Wang JM
        • Rogers TJ
        • Caterina M
        • Oppenheim JJ
        A proinflammatory chemokine, CCL3, sensitizes the heat- and capsaicin-gated ion channel TRPV1.
        Proc Natl Acad Sci U S A. 2005; 102: 4536-4541
        • Zhang W
        • Yu Y
        • Hertwig F
        • Thierry-Mieg J
        • Zhang W
        • Thierry-Mieg D
        • Wang J
        • Furlanello C
        • Devanarayan V
        • Cheng J
        • Deng Y
        • Hero B
        • Hong H
        • Jia M
        • Li L
        • Lin SM
        • Nikolsky Y
        • Oberthuer A
        • Qing T
        • Su Z
        • Volland R
        • Wang C
        • Wang MD
        • Ai J
        • Albanese D
        • Asgharzadeh S
        • Avigad S
        • Bao W
        • Bessarabova M
        • Brilliant MH
        • Brors B
        • Chierici M
        • Chu TM
        • Zhang J
        • Grundy RG
        • He MM
        • Hebbring S
        • Kaufman HL
        • Lababidi S
        • Lancashire LJ
        • Li Y
        • Lu XX
        • Luo H
        • Ma X
        • Ning B
        • Noguera R
        • Peifer M
        • Phan JH
        • Roels F
        • Rosswog C
        • Shao S
        • Shen J
        • Theissen J
        • Tonini GP
        • Vandesompele J
        • Wu PY
        • Xiao W
        • Xu J
        • Xu W
        • Xuan J
        • Yang Y
        • Ye Z
        • Dong Z
        • Zhang KK
        • Yin Y
        • Zhao C
        • Zheng Y
        • Wolfinger RD
        • Shi T
        • Malkas LH
        • Berthold F
        • Wang J
        • Tong W
        • Shi L
        • Peng Z
        • Fischer M
        Comparison of RNA-seq and microarray-based models for clinical endpoint prediction.
        Genome Biol. 2015; 16: 133