Engineered induced-pluripotent stem cell derived monocyte extracellular vesicles alter inflammation in HIV humanized mice

Abstract

Aim: A peripheral inflammatory response can drive neuroinflammation in a number of infections including human immunodeficiency virus (HIV). Monocyte/macrophage (M/Mφ) activation is a hallmark of acute HIV infection and a source of chronic inflammation in a subset of HIV-infected individuals. We sought to decrease peripheral inflammation and M/Mφ transmigration after HIV infection by engineering extracellular vesicles (EV) to antagonize a microRNA (miR) associated with inflammation. We hypothesized that induced pluripotent stem cell (iPSC)-derived monocyte EVs (mEVs), engineered to contain an antagomir to miR-155 (αmiR mEV) would target monocyte inflammation and influence neuroinflammation in an HIV-infected humanized mice.

Methods: mEVs were characterized by tetraspanins, nanoparticle tracking analysis, electron microscopy, and their preferential entry into circulating monocytes as well as testing for endogenous selected miRNAs. HIV-infected humanized mice were treated with control or antagomir155 mEVs. Plasma viral load was measured plus activation markers on lymphocytes and monocytes and the number of macrophages in the brain was quantified.

Results: mEVs preferentially entered peripheral monocytes. HIV infection increased C-C chemokine receptor type 5 (CCR5) and major histocompatibility complex, class II, DR (HLA-DR) expression on T cells and monocytes. Treatments with mEVs did not decrease plasma HIV viral load; however, mEVs alone resulted in a decrease in %CCR5+ and %HLA-DR+ on T cells and an increase in %CCR5+ monocytes. αmiR mEVs decreased %CCR5 on M/Mφ. The mEV-treated HIV-infected mice did not show an increase in macrophage transmigration to the brain.

Conclusion: mEVs alone caused an unexpected decrease in lymphocyte activation and increase in monocyte %CCR5; however, this did not translate to an increase in macrophage transmigration to the brain.

Keywords

Monocyte / extracellular vesicles / pluripotent stem cells / miRNAs / HIV / bone marrow-liver-thymic (BLT) mouse / inflammation / neuroinflammation

References

[1] Gallitano SM,Brar K.Use of tumor necrosis factor (TNF) inhibitors in patients with HIV/AIDS.J Am Acad Dermatol2016;74:974-80

[2] Kumar A,Herbein G.Targeting TNF-alpha in HIV-1 infection.Curr Drug Targets2016;17:15-22

[3] Bochnakian A,Zisoulis DG.Interferon-inducible microRNA miR-128 modulates HIV-1 replication by targeting TNPO3 mRNA.J Virol2019;93:e00364-19 PMCID:PMC6798100

[4] Tang N,Gupta A,Pulliam L.Monocyte exosomes induce adhesion molecules and cytokines via activation of NF-κB in endothelial cells.FASEB J2016;30:3097-106 PMCID:PMC5001509

[5] Dalvi P,Tang N.Immune activated monocyte exosomes alter microRNAs in brain endothelial cells and initiate an inflammatory response through the TLR4/MyD88 pathway.Sci Rep2017;7:9954 PMCID:PMC5577170

[6] Elton TS,Elton SM.Regulation of the MIR155 host gene in physiological and pathological processes.Gene2013;532:1-12

[7] Akkina R,Balazs AB.Improvements and limitations of humanized mouse models for HIV research: NIH/NIAID “meet the experts” 2015 workshop summary.AIDS Res Hum Retroviruses2016;32:109-19

[8] Carrillo MA,Kitchen SG.The use of the humanized mouse model in gene therapy and immunotherapy for HIV and cancer.Front Immunol2018;9:746 PMCID:PMC5932400

[9] Zhen A,Youn C.Targeting type I interferon-mediated activation restores immune function in chronic HIV infection.J Clin Invest2017;127:260-8 PMCID:PMC5199686

[10] Kitchen SG,Bristol G.In vivo suppression of HIV by antigen specific T cells derived from engineered hematopoietic stem cells.PLoS Pathog2012;8:e1002649 PMCID:PMC3325196

[11] Rempel H,Calosing C,Pulliam L.Interferon-alpha drives monocyte gene expression in chronic unsuppressed HIV-1 infection.AIDS2010;24:1415-23 PMCID:PMC2991092

[12] Volarevic V,Gazdic M.Ethical and safety issues of stem cell-based therapy.Int J Med Sci2018;15:36-45 PMCID:PMC5765738

[13] Harrell CR,Djonov V,Volarevic V.Mesenchymal stem cell-derived exosomes and other extracellular vesicles as new remedies in the therapy of inflammatory diseases.Cells2019;8:1605 PMCID:PMC6952783

[14] Duan L,Zhao X.Extracellular vesicles derived from human placental mesenchymal stem cells alleviate experimental colitis in mice by inhibiting inflammation and oxidative stress.Int J Mol Med2020;46:1551-61 PMCID:PMC7447323

[15] Feng R,Chen K,Lin Y.Stem cell-derived extracellular vesicles mitigate ageing-associated arterial stiffness and hypertension.J Extracell Vesicles2020;9:1783869 PMCID:PMC7480600

[16] Fang SB,Meng XC.Small extracellular vesicles derived from human MSCs prevent allergic airway inflammation via immunomodulation on pulmonary macrophages.Cell Death Dis2020;11:409 PMCID:PMC7264182

[17] Khan M,Abramova T.Embryonic stem cell-derived exosomes promote endogenous repair mechanisms and enhance cardiac function following myocardial infarction.Circ Res2015;117:52-64 PMCID:PMC4482130

[18] Huang T,Fang L.Extracellular vesicle-derived miR-511-3p from hypoxia preconditioned adipose mesenchymal stem cells ameliorates spinal cord injury through the TRAF6/S1P axis.Brain Res Bull2022;180:73-85

[19] Qian Y,Chen L.Mesenchymal Stem cell-derived extracellular vesicles alleviate M1 microglial activation in brain injury of mice with subarachnoid hemorrhage via microRNA-140-5p delivery.Int J Neuropsychopharmacol2022;25:328-38

[20] Song BW,Kim R.Multiplexed targeting of miRNA-210 in stem cell-derived extracellular vesicles promotes selective regeneration in ischemic hearts.Exp Mol Med2021;53:695-708 PMCID:PMC8102609

[21] Rozier P,Maria ATJ.Mesenchymal stromal cells-derived extracellular vesicles alleviate systemic sclerosis via miR-29a-3p.J Autoimmun2021;121:102660

[22] Alivernini S,McSharry C.MicroRNA-155-at the critical interface of innate and adaptive immunity in arthritis.Front Immunol2017;8:1932 PMCID:PMC5760508

[23] Jablonski KA,Amici SA,Guerau-de-Arellano M.Control of the inflammatory macrophage transcriptional signature by miR-155.PLoS One2016;11:e0159724 PMCID:PMC4957803

[24] Gaudet AD,Hall JC.miR-155 deletion in mice overcomes neuron-intrinsic and neuron-extrinsic barriers to spinal cord repair.J Neurosci2016;36:8516-32 PMCID:PMC4978808

[25] Gaudet AD,Ayala MT,Watkins LR.Aging and miR-155 in mice influence survival and neuropathic pain after spinal cord injury.Brain Behav Immun2021;97:365-70 PMCID:PMC8453092

[26] Huang Q,Luo M.HIV-1-induced miR-146a Attenuates monocyte migration by targeting CCL5 in Human primary macrophages.AIDS Res Hum Retroviruses2018;34:580-9

[27] Bhargavan B,McMillan JE.CCR5 antagonist reduces HIV-induced amyloidogenesis, tau pathology, neurodegeneration, and blood-brain barrier alterations in HIV-infected hu-PBL-NSG mice.Mol Neurodegener2021;16:78 PMCID:PMC8607567

[28] Zitzer NC,Meng X.MicroRNA-155 modulates acute graft-versus-host disease by impacting T cell expansion, migration, and effector function.J Immunol2018;200:4170-9 PMCID:PMC5988971

[29] Zang YC,Halder JB.Aberrant T cell migration toward RANTES and MIP-1 alpha in patients with multiple sclerosis. Overexpression of chemokine receptor CCR5.Brain2000;123:1874-82

[30] Buckner CM,Willams DW,Berman JW.Characterization of monocyte maturation/differentiation that facilitates their transmigration across the blood-brain barrier and infection by HIV: implications for NeuroAIDS.Cell Immunol2011;267:109-23 PMCID:PMC4335637

[31] Ndhlovu LC,Chew GM.Treatment intensification with maraviroc (CCR5 antagonist) leads to declines in CD16-expressing monocytes in cART-suppressed chronic HIV-infected subjects and is associated with improvements in neurocognitive test performance: implications for HIV-associated neurocognitive disease (HAND).J Neurovirol2014;20:571-82 PMCID:PMC4268390

[32] Pulliam L,Rempel H.Invasive chronic inflammatory monocyte phenotype in subjects with high HIV-1 viral load.J Neuroimmunol2004;157:93-8

[33] Veenstra M,Li M,Clements JE.Mechanisms of CNS viral seeding by HIV⁺ CD14⁺ CD16⁺ monocytes: establishment and reseeding of viral reservoirs contributing to HIV-associated neurocognitive disorders.mBio2017;8:e01280-17 PMCID:PMC5654927

[34] Guilliams M,Yona S.Developmental and functional heterogeneity of monocytes.Immunity2018;49:595-613

[35] Zhuang X,Grizzle W.Treatment of brain inflammatory diseases by delivering exosome encapsulated anti-inflammatory drugs from the nasal region to the brain.Mol Ther2011;19:1769-79 PMCID:PMC3188748