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Polystyrene microplastics induce an immunometabolic active state in macrophages

Abstract

Anti-inflammatory and proinflammatory responses in macrophages are influenced by cellular metabolism. Macrophages are the primary phagocyte in mucosal environments (i.e., intestinal tract and lungs) acting as first-line defense against microorganisms and environmental pollutants. Given the extensive contamination of our food and water sources with microplastics, we aimed to examine the metabolic response in macrophages to microplastic particles (MPs). Utilizing murine macrophages, we assessed the metabolic response of macrophages after polystyrene MP phagocytosis. The phagocytosis of MP by macrophages induced a metabolic shift toward glycolysis and a reduction in mitochondrial respiration that was associated with an increase of cell surface markers CD80 and CD86 and cytokine gene expression associated with glycolysis. The gastrointestinal consequences of this metabolic switch in the context of an immune response remain uncertain, but the global rise of plastic pollution and MP ingestion potentially poses an unappreciated health risk.

Graphical abstract

Macrophage phagocytosis of microplastics alters cellular metabolism.

- Macrophages cannot degrade PS MP.

- MP phagocytosis increases glycolysis in murine macrophages.

- MP phagocytosis reduces mitochondrial respiration in murine macrophages.

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Fig. 1
Fig. 2

Data availability

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Code availability

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Abbreviations

ECAR:

Extracellular acidification rate

GRA:

Glycolytic rate assay

IBD:

Inflammatory bowel disease

IEC:

Intestinal epithelial cell

LAP:

LC3-associated phagocytosis

MFI:

Mean fluorescence intensity

mitoOCR:

Mitochondrial oxygen consumption rate

MP:

Microplastic particles

PER:

Proton efflux rate

TCA:

Tricarboxylic acid cycle

References

  • Bain CC, Schridde A. Origin, differentiation, and function of intestinal macrophages. Front Immunol. 2018;9:2733.

    PubMed  PubMed Central  Google Scholar 

  • Bain CC, Scott CL, Uronen-Hansson H, Gudjonsson S, Jansson O, Grip O, Guilliams M, Malissen B, Agace WW, Mowat AM. Resident and pro-inflammatory macrophages in the colon represent alternative context-dependent fates of the same Ly6Chi monocyte precursors. Mucosal Immunol. 2013;6:498–510.

    CAS  PubMed  Google Scholar 

  • Castillo EF, Dekonenko A, Arko-Mensah J, Mandell MA, Dupont N, Jiang S, Delgado-Vargas M, Timmins GS, Bhattacharya D, Yang H, et al. Autophagy protects against active tuberculosis by suppressing bacterial burden and inflammation. Proc Natl Acad Sci U S A. 2012;109:E3168-3176.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Catarino AI, Macchia V, Sanderson WG, Thompson RC, Henry TB. Low levels of microplastics (MP) in wild mussels indicate that MP ingestion by humans is minimal compared to exposure via household fibres fallout during a meal. Environ Pollut. 2018;237:675–84.

    CAS  PubMed  Google Scholar 

  • Cau A, Avio CG, Dessi C, Follesa MC, Moccia D, Regoli F, Pusceddu A. Microplastics in the crustaceans Nephrops norvegicus and Aristeus antennatus: flagship species for deep-sea environments? Environ Pollut. 2019;255:113107.

    CAS  PubMed  Google Scholar 

  • Cheung LTO, Lui CY, Fok L. Microplastic contamination of wild and captive flathead grey mullet (Mugil cephalus). Int J Environ Res Public Health. 2018;15(4):597. https://doi.org/10.3390/ijerph15040597.

    CAS  Article  PubMed Central  Google Scholar 

  • Cho Y, Shim WJ, Jang M, Han GM, Hong SH. Abundance and characteristics of microplastics in market bivalves from South Korea. Environ Pollut. 2019;245:1107–16.

    CAS  PubMed  Google Scholar 

  • Cox KD, Covernton GA, Davies HL, Dower JF, Juanes F, Dudas SE. Human consumption of microplastics. Environ Sci Technol. 2019;53:7068–74.

    CAS  PubMed  Google Scholar 

  • Deng Y, Zhang Y, Lemos B, Ren H. Tissue accumulation of microplastics in mice and biomarker responses suggest widespread health risks of exposure. Sci Rep. 2017;7:46687.

    PubMed  PubMed Central  Google Scholar 

  • Diskin C, Palsson-McDermott EM. Metabolic modulation in macrophage effector function. Front Immunol. 2018;9:270.

    PubMed  PubMed Central  Google Scholar 

  • Doyen P, Hermabessiere L, Dehaut A, Himber C, Decodts M, Degraeve T, Delord L, Gaboriaud M, Mone P, Sacco J, et al. Occurrence and identification of microplastics in beach sediments from the Hauts-de-France region. Environ Sci Pollut Res Int. 2019;26:28010–21.

    CAS  PubMed  Google Scholar 

  • Feng Z, Zhang T, Li Y, He X, Wang R, Xu J, Gao G. The accumulation of microplastics in fish from an important fish farm and mariculture area, Haizhou Bay China. Sci Total Environ. 2019;696:133948.

    CAS  PubMed  Google Scholar 

  • Florey O, Gammoh N, Kim SE, Jiang X, Overholtzer M. V-ATPase and osmotic imbalances activate endolysosomal LC3 lipidation. Autophagy. 2015;11:88–99.

    PubMed  Google Scholar 

  • Galluzzi L, Baehrecke EH, Ballabio A, Boya P, Bravo-San Pedro JM, Cecconi F, Choi AM, Chu CT, Codogno P, Colombo MI, et al. Molecular definitions of autophagy and related processes. EMBO J. 2017;36:1811–36.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Gao Y, Liu Y, Hong L, Yang Z, Cai X, Chen X, Fu Y, Lin Y, Wen W, Li S, et al. Golgi-associated LC3 lipidation requires V-ATPase in noncanonical autophagy. Cell Death Dis. 2016;7:e2330.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Gatica D, Lahiri V, Klionsky DJ. Cargo recognition and degradation by selective autophagy. Nat Cell Biol. 2018;20:233–42.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Hanada T, Noda NN, Satomi Y, Ichimura Y, Fujioka Y, Takao T, Inagaki F, Ohsumi Y. The Atg12-Atg5 conjugate has a novel E3-like activity for protein lipidation in autophagy. J Biol Chem. 2007;282:37298–302.

    CAS  PubMed  Google Scholar 

  • Heckmann BL, Teubner BJW, Tummers B, Boada-Romero E, Harris L, Yang M, Guy CS, Zakharenko SS, Green DR. LC3-associated endocytosis facilitates beta-amyloid clearance and mitigates neurodegeneration in murine Alzheimer’s disease. Cell. 2019;178(536–551):e514.

    Google Scholar 

  • Hu Y, Mai W, Chen L, Cao K, Zhang B, Zhang Z, Liu Y, Lou H, Duan S, Gao Z. mTOR-mediated metabolic reprogramming shapes distinct microglia functions in response to lipopolysaccharide and ATP. Glia. 2020;68:1031–45.

    PubMed  Google Scholar 

  • Jacquin E, Leclerc-Mercier S, Judon C, Blanchard E, Fraitag S, Florey O. Pharmacological modulators of autophagy activate a parallel noncanonical pathway driving unconventional LC3 lipidation. Autophagy. 2017;13:854–67.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Jahan S, Strezov V, Weldekidan H, Kumar R, Kan T, Sarkodie SA, He J, Dastjerdi B, Wilson SP. Interrelationship of microplastic pollution in sediments and oysters in a seaport environment of the eastern coast of Australia. Sci Total Environ. 2019;695:133924.

    CAS  PubMed  Google Scholar 

  • Jha AK, Huang SC, Sergushichev A, Lampropoulou V, Ivanova Y, Loginicheva E, Chmielewski K, Stewart KM, Ashall J, Everts B, et al. Network integration of parallel metabolic and transcriptional data reveals metabolic modules that regulate macrophage polarization. Immunity. 2015;42:419–30.

    CAS  PubMed  Google Scholar 

  • Jin Y, Lu L, Tu W, Luo T, Fu Z. Impacts of polystyrene microplastic on the gut barrier, microbiota and metabolism of mice. Sci Total Environ. 2019;649:308–17.

    CAS  PubMed  Google Scholar 

  • Karami A, Golieskardi A, Ho YB, Larat V, Salamatinia B. Microplastics in eviscerated flesh and excised organs of dried fish. Sci Rep. 2017a;7:5473.

    PubMed  PubMed Central  Google Scholar 

  • Karami A, Golieskardi A, Keong Choo C, Larat V, Galloway TS, Salamatinia B. The presence of microplastics in commercial salts from different countries. Sci Rep. 2017b;7:46173.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kim JS, Lee HJ, Kim SK, Kim HJ. Global pattern of microplastics (MPs) in commercial food-grade salts: sea salt as an indicator of seawater MP pollution. Environ Sci Technol. 2018;52:12819–28.

    CAS  PubMed  Google Scholar 

  • Knoop KA, Miller MJ, Newberry RD. Transepithelial antigen delivery in the small intestine: different paths, different outcomes. Curr Opin Gastroenterol. 2013;29:112–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Koelmans AA, Mohamed Nor NH, Hermsen E, Kooi M, Mintenig SM, De France J. Microplastics in freshwaters and drinking water: critical review and assessment of data quality. Water Res. 2019;155:410–22.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lee HY, Kim J, Quan W, Lee JC, Kim MS, Kim SH, Bae JW, Hur KY, Lee MS. Autophagy deficiency in myeloid cells increases susceptibility to obesity-induced diabetes and experimental colitis. Autophagy. 2016;12:1390–403.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Li J, Yang D, Li L, Jabeen K, Shi H. Microplastics in commercial bivalves from China. Environ Pollut. 2015;207:190–5.

    CAS  PubMed  Google Scholar 

  • Litvak Y, Byndloss MX, Bäumler AJ. Colonocyte metabolism shapes the gut microbiota. Science. 2018;362(6418):eaat9076. https://doi.org/10.1126/science.aat9076.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Lu L, Wan Z, Luo T, Fu Z, Jin Y. Polystyrene microplastics induce gut microbiota dysbiosis and hepatic lipid metabolism disorder in mice. Sci Total Environ. 2018;631–632:449–58.

    PubMed  Google Scholar 

  • Ma TY, Nguyen D, Bui V, Nguyen H, Hoa N. Ethanol modulation of intestinal epithelial tight junction barrier. Am J Physiol. 1999;276:G965-974.

    CAS  PubMed  Google Scholar 

  • Martinez J, Verbist K, Wang R, Green DR. The relationship between metabolism and the autophagy machinery during the innate immune response. Cell Metab. 2013;17:895–900.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Martinez J, Malireddi RK, Lu Q, Cunha LD, Pelletier S, Gingras S, Orchard R, Guan JL, Tan H, Peng J, et al. Molecular characterization of LC3-associated phagocytosis reveals distinct roles for Rubicon, NOX2 and autophagy proteins. Nat Cell Biol. 2015;17:893–906.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Mazzini E, Massimiliano L, Penna G, Rescigno M. Oral tolerance can be established via gap junction transfer of fed antigens from CX3CR1(+) macrophages to CD103(+) dendritic cells. Immunity. 2014;40:248–61.

    CAS  PubMed  Google Scholar 

  • Merkley SD, Goodfellow SM, Guo Y, Wilton ZER, Byrum JR, Schwalm KC, Dinwiddie DL, Gullapalli RR, Deretic V, Jimenez Hernandez A, Bradfute SB, In JG, Castillo EF. Non-autophagy role of Atg5 and NBR1 in unconventional secretion of IL-12 prevents gut dysbiosis and inflammation bioRxiv 2020. https://doi.org/10.1101/2020.12.07.414227.

  • Millet P, Vachharajani V, McPhail L, Yoza B, McCall CE. GAPDH Binding to TNF-alpha mRNA contributes to posttranscriptional repression in monocytes: a novel mechanism of communication between inflammation and metabolism. J Immunol. 2016;196:2541–51.

    CAS  PubMed  Google Scholar 

  • Mizushima N, Noda T, Yoshimori T, Tanaka Y, Ishii T, George MD, Klionsky DJ, Ohsumi M, Ohsumi Y. A protein conjugation system essential for autophagy. Nature. 1998;395:395–8.

    CAS  PubMed  Google Scholar 

  • Mogilenko DA, Haas JT, L’Homme L, Fleury S, Quemener S, Levavasseur M, Becquart C, Wartelle J, Bogomolova A, Pineau L, et al. Metabolic and innate immune cues merge into a specific inflammatory response via the UPR. Cell. 2019;178:263.

    CAS  PubMed  Google Scholar 

  • Nan B, Su L, Kellar C, Craig NJ, Keough MJ, Pettigrove V. Identification of microplastics in surface water and Australian freshwater shrimp Paratya australiensis in Victoria Australia. Environ Pollut. 2020;259:113865.

    CAS  PubMed  Google Scholar 

  • Newsholme P, Gordon S, Newsholme EA. Rates of utilization and fates of glucose, glutamine, pyruvate, fatty acids and ketone bodies by mouse macrophages. Biochem J. 1987;242:631–6.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Ng SC, Shi HY, Hamidi N, Underwood FE, Tang W, Benchimol EI, Panaccione R, Ghosh S, Wu JCY, Chan FKL, et al. Worldwide incidence and prevalence of inflammatory bowel disease in the 21st century: a systematic review of population-based studies. Lancet. 2018;390:2769–78.

    Google Scholar 

  • Niess JH, Brand S, Gu X, Landsman L, Jung S, McCormick BA, Vyas JM, Boes M, Ploegh HL, Fox JG, et al. CX3CR1-mediated dendritic cell access to the intestinal lumen and bacterial clearance. Science. 2005;307:254–8.

    CAS  PubMed  Google Scholar 

  • Noda NN, Fujioka Y, Hanada T, Ohsumi Y, Inagaki F. Structure of the Atg12-Atg5 conjugate reveals a platform for stimulating Atg8-PE conjugation. EMBO Rep. 2013;14:206–11.

    CAS  PubMed  Google Scholar 

  • Novotna K, Cermakova L, Pivokonska L, Cajthaml T, Pivokonsky M. Microplastics in drinking water treatment - current knowledge and research needs. Sci Total Environ. 2019;667:730–40.

    CAS  PubMed  Google Scholar 

  • Palsson-McDermott EM, Dyck L, Zaslona Z, Menon D, McGettrick AF, Mills KHG, O’Neill LA. Pyruvate kinase M2 is required for the expression of the immune checkpoint PD-L1 in immune cells and tumors. Front Immunol. 2017;8:1300.

    PubMed  PubMed Central  Google Scholar 

  • Ramsperger AFRM, Narayana VKB, Gross W, Mohanraj J, Thelakkat M, Greiner A, Schmalz H, Kress H, Laforsch C. Environmental exposure enhances the internalization of microplastic particles into cells. Sci Adv. 2020;6(50):eabd1211. https://doi.org/10.1126/sciadv.abd1211.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Rummel CD, Loder MG, Fricke NF, Lang T, Griebeler EM, Janke M, Gerdts G. Plastic ingestion by pelagic and demersal fish from the North Sea and Baltic Sea. Mar Pollut Bull. 2016;102:134–41.

    CAS  PubMed  Google Scholar 

  • Sakoh-Nakatogawa M, Matoba K, Asai E, Kirisako H, Ishii J, Noda NN, Inagaki F, Nakatogawa H, Ohsumi Y. Atg12-Atg5 conjugate enhances E2 activity of Atg3 by rearranging its catalytic site. Nat Struct Mol Biol. 2013;20:433–9.

    CAS  PubMed  Google Scholar 

  • Sanjuan MA, Dillon CP, Tait SW, Moshiach S, Dorsey F, Connell S, Komatsu M, Tanaka K, Cleveland JL, Withoff S, et al. Toll-like receptor signalling in macrophages links the autophagy pathway to phagocytosis. Nature. 2007;450:1253–7.

    CAS  PubMed  Google Scholar 

  • Schnyder J, Baggiolini M. Role of phagocytosis in the activation of macrophages. J Exp Med. 1978;148:1449–57.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Schwabl P, Koppel S, Konigshofer P, Bucsics T, Trauner M, Reiberger T, Liebmann B. Detection of various microplastics in human stool: a prospective case series. Ann Intern Med. 2019;171:453–7.

    PubMed  Google Scholar 

  • Shanmugam N, Reddy MA, Guha M, Natarajan R. High glucose-induced expression of proinflammatory cytokine and chemokine genes in monocytic cells. Diabetes. 2003;52:1256–64.

    CAS  PubMed  Google Scholar 

  • Stock V, Bohmert L, Lisicki E, Block R, Cara-Carmona J, Pack LK, Selb R, Lichtenstein D, Voss L, Henderson CJ, et al. Uptake and effects of orally ingested polystyrene microplastic particles in vitro and in vivo. Arch Toxicol. 2019;93:1817–33.

    CAS  PubMed  Google Scholar 

  • Tanaka K, Takada H. Microplastic fragments and microbeads in digestive tracts of planktivorous fish from urban coastal waters. Sci Rep. 2016;6:34351.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Viola A, Munari F, Sanchez-Rodriguez R, Scolaro T, Castegna A. The metabolic signature of macrophage responses. Front Immunol. 2019;10:1462.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Wang X, Rao H, Zhao J, Wee A, Li X, Fei R, Huang R, Wu C, Liu F, Wei L. STING expression in monocyte-derived macrophages is associated with the progression of liver inflammation and fibrosis in patients with nonalcoholic fatty liver disease. Lab Invest. 2020;100:542–52.

    CAS  PubMed  Google Scholar 

  • Wright SL, Kelly FJ. Plastic and human health: a micro issue? Environ Sci Technol. 2017;51:6634–47.

    CAS  PubMed  Google Scholar 

  • Wu F, Wang Y, Leung JYS, Huang W, Zeng J, Tang Y, Chen J, Shi A, Yu X, Xu X, et al. Accumulation of microplastics in typical commercial aquatic species: a case study at a productive aquaculture site in China. Sci Total Environ. 2020;708:135432.

    CAS  PubMed  Google Scholar 

  • Wu MY, Lu JH. Autophagy and Macrophage Functions: Inflammatory Response and Phagocytosis. Cells. 2019;9(1):70. https://doi.org/10.3390/cells9010070.

    CAS  Article  PubMed Central  Google Scholar 

  • Xie M, Yu Y, Kang R, Zhu S, Yang L, Zeng L, Sun X, Yang M, Billiar TR, Wang H, et al. PKM2-dependent glycolysis promotes NLRP3 and AIM2 inflammasome activation. Nat Commun. 2016;7:13280.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Yoshimori T, Yamamoto A, Moriyama Y, Futai M, Tashiro Y. Bafilomycin A1, a specific inhibitor of vacuolar-type H(+)-ATPase, inhibits acidification and protein degradation in lysosomes of cultured cells. J Biol Chem. 1991;266:17707–12.

    CAS  PubMed  Google Scholar 

  • Zeng Z, Shi YX, Tsao T, Qiu Y, Kornblau SM, Baggerly KA, Liu W, Jessen K, Liu Y, Kantarjian H, et al. Targeting of mTORC1/2 by the mTOR kinase inhibitor PP242 induces apoptosis in AML cells under conditions mimicking the bone marrow microenvironment. Blood. 2012;120:2679–89.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang H, Zheng L, McGovern DP, Hamill AM, Ichikawa R, Kanazawa Y, Luu J, Kumagai K, Cilluffo M, Fukata M, et al. Myeloid ATG16L1 facilitates host-bacteria interactions in maintaining intestinal homeostasis. J Immunol. 2017;198:2133–46.

    CAS  PubMed  Google Scholar 

  • Zhao M, Burisch J. Impact of genes and the environment on the pathogenesis and disease course of inflammatory bowel disease. Dig Dis Sci. 2019;64:1759–69.

    CAS  PubMed  Google Scholar 

  • Zhao Z, Fux B, Goodwin M, Dunay IR, Strong D, Miller BC, Cadwell K, Delgado MA, Ponpuak M, Green KG, et al. Autophagosome-independent essential function for the autophagy protein Atg5 in cellular immunity to intracellular pathogens. Cell Host Microbe. 2008;4:458–69.

    CAS  PubMed  PubMed Central  Google Scholar 

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Funding

This work was supported in part by the National Center for Research Resources and the National Center for Advancing Translational Sciences of the National Institutes of Health (NIH) through grant no. UL1TR001449 (EFC), in part by NIH grant 1R56ES032037-01 (E.F.C.), in part by NIH grant P20GM121176 (EFC), P20GM130422 (MJC), and New Mexico Medical Trust C-2446-RAC (EFC). SMG was supported in part by the Infectious Disease and Inflammation Program Pre-doctoral T32 training grant, NIH/NIAID grant T32AI007538.

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SDM participated in writing the manuscript and performed all experiments with help from HCM, JMH, CLL, and JW. SMG performed confocal microscopy. MJC provided critical reagents and helped design the study. EFC designed the study, analyzed data, and wrote the paper. All authors approved the final version of the manuscript.

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Correspondence to Eliseo F. Castillo.

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All experiments were approved by the Institutional Animal Care and Use Committee of the University of New Mexico Health Sciences Center, in accordance with the National Institutes of Health guidelines for use of live animals. The University of New Mexico Health Sciences Center is accredited by the American Association for Accreditation of Laboratory Animal Care.

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Merkley, S.D., Moss, H.C., Goodfellow, S.M. et al. Polystyrene microplastics induce an immunometabolic active state in macrophages. Cell Biol Toxicol 38, 31–41 (2022). https://doi.org/10.1007/s10565-021-09616-x

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Keywords

  • Microplastics
  • Macrophage
  • Metabolism
  • Inflammation
  • Particulates