PreImplantation factor prevents atherosclerosis via its immunomodulatory effects without affecting serum lipids

 

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Summary

PreImplantation factor (PIF) is a 15-amino acid peptide endogenously secreted by viable embryos, regulating/enabling maternal (host) acceptance/tolerance to the “invading” embryo (allograft) all-while preserving maternal immunity to fight infections. Such attributes make PIF a potential therapeutic agent for chronic inflammatory diseases. We investigated whether PIF’s immunomodulatory properties prevent progression of atherosclerosis in the hyper-cholesterolaemic ApoEdeficient murine model. Male, high-fat diet fed, ApoE-deficient (ApoE^-/-) mice were administered either PBS, scrambled PIF (0.3–3 mg/kg) or PIF (0.3–3 mg/kg) for seven weeks. After treatment, PIF (3 mg/kg)-treated ApoE^-/- mice displayed significantly reduced atherosclerosis lesion burden in the aortic sinus and aortic arch, without any effect on lipid profile. PIF also caused a significant reduction in infiltration of macrophages, decreased expression of pro-inflammatory adhesion molecules, cytokines and chemokines in the plaque, and reduced circulating IFN-γ levels. PIF preferentially binds to monocytes/neutrophils. In vitro, PIF attenuated monocyte migration (MCP-1-induced chemotaxis assay) and in vivo in LPS peritonitis model. Also PIF prevented leukocyte extravasation (peritonitis thioglycollate-induced model), demonstrating that PIF exerts its effect in part by modulation of monocyte function. Inhibition of the potassium channel KCNAB3 (Kv1.3) and of the insulin degrading enzyme (IDE) was demonstrated as potential mechanism of PIF’s immunomodulatory effects. In conclusion, PIF regulates/lowers inflammation and prevents atherosclerosis development without affecting circulating lipids. Overall our findings establish PIF as a strong immunomodulatory drug candidate for atherosclerosis therapy.

Supplementary Material to this article is available online at www.thrombosis-online.com.

^* YCC and JF contributed equally

^# ERB and KP contributed equally.

• References

• 1 Libby P, Lichtman AH, Hansson GK. Immune effector mechanisms implicated in atherosclerosis: from mice to humans. Immunity 2013; 38: 1092-1104.
  CrossrefPubMedGoogle Scholar
• 2 Randolph GJ. Mechanisms that regulate macrophage burden in atherosclerosis. Circ Res 2014; 114: 1757-1771.
  CrossrefPubMedGoogle Scholar
• 3 Weber C, Noels H. Atherosclerosis: current pathogenesis and therapeutic options. Nat Med 2011; 17: 1410-1422.
  CrossrefPubMedGoogle Scholar
• 4 Ramu S, Stamatkin C, Timms L. et al. PreImplantation factor (PIF) detection in maternal circulation in early pregnancy correlates with live birth (bovine model). Reprod Biol Endocrinol 2013; 11: 105.
  CrossrefPubMedGoogle Scholar
• 5 Stamatkin CW, Roussev RG, Stout M. et al. Preimplantation factor negates embryo toxicity and promotes embryo development in culture. Reprod Biomed Online 2011; 23: 517-524.
  CrossrefPubMedGoogle Scholar
• 6 Barnea ER, Kirk D, Paidas MJ. PreImplantation factor (PIF) promoting role in embryo implantation: increases endometrial Integrin-alpha2beta3, amphiregulin and epiregulin while reducing betacellulin expression via MAPK in decidua. Reprod Biol Endocrinol 2012; 10: 50.
  CrossrefPubMedGoogle Scholar
• 7 Paidas MJ, Annunziato J, Romano M. et al. Pregnancy and multiple sclerosis (MS): a beneficial association. Possible therapeutic application of embryo-specific pre-implantation factor (PIF*). Am J Reprod Immunol 2012; 68: 456-464.
  CrossrefPubMedGoogle Scholar
• 8 Barnea ER, Lahijani KI, Roussev R. et al. Use of lymphocyte platelet binding assay for detecting a preimplantation factor: a quantitative assay. Am J Reprod Immunol 1994; 32: 133-138.
  CrossrefPubMedGoogle Scholar
• 9 Roussev RG, Coulam CB, Kaider BD. et al. Embryonic origin of preimplantation factor (PIF): biological activity and partial characterization. Mol Hum Reprod 1996; 2: 883-887.
  CrossrefPubMedGoogle Scholar
• 10 Stamatkin CW, Roussev RG, Stout M. et al. PreImplantation Factor (PIF) correlates with early mammalian embryo development-bovine and murine models. Reprod Biol Endocrinol 2011; 9: 63.
  CrossrefPubMedGoogle Scholar
• 11 Paidas MJ, Krikun G, Huang SJ. et al. A genomic and proteomic investigation of the impact of preimplantation factor on human decidual cells. Am J Obstet Gynecol 2010; 202: 459 e451-458.
  PubMedGoogle Scholar
• 12 Barnea ER, Kirk D, Paidas MJ. Preimplantation Factor (PIF*) promoting role in embryo implantation: increases endometrial Integrin-alpha2beta3 and amphiregulin and epiregulin while reducing betacellulin expression via MAPK in decidua. Reprod Biol Endocrinol 2012; 10: 50.
  CrossrefPubMedGoogle Scholar
• 13 Duzyj CM, Barnea ER, Li M. et al. Preimplantation factor promotes first trimester trophoblast invasion. Am J Obstet Gynecol 2010; 203: 402 e401-404.
  PubMedGoogle Scholar
• 14 Rosario GX, Modi DN, Sachdeva G. et al. Morphological events in the primate endometrium in the presence of a preimplantation embryo, detected by the serum preimplantation factor bioassay. Hum Reprod 2005; 20: 61-71.
  CrossrefPubMedGoogle Scholar
• 15 Barnea ER. Insight into early pregnancy events: the emerging role of the embryo. Am J Reprod Immunol 2004; 51: 319-322.
  CrossrefPubMedGoogle Scholar
• 16 Roussev RG, Dons’koi BV, Stamatkin C. et al. Preimplantation factor inhibits circulating natural killer cell cytotoxicity and reduces CD69 expression: implications for recurrent pregnancy loss therapy. Reprod Biomed Online 2013; 26: 79-87.
  CrossrefPubMedGoogle Scholar
• 17 Barnea ER, Kirk D, Ramu S. et al. PreImplantation Factor (PIF) orchestrates systemic antiinflammatory response by immune cells: effect on peripheral blood mononuclear cells. Am J Obstet Gynecol 2012; 207: 313 e311-311.
  PubMedGoogle Scholar
• 18 Weiss L, Or R, Jones RC. et al. Preimplantation factor (PIF*) reverses neuroinflammation while promoting neural repair in EAE model. J Neurol Sci 2012; 312: 146-157.
  CrossrefPubMedGoogle Scholar
• 19 Weiss L, Bernstein S, Jones R. et al. Preimplantation factor (PIF) analog prevents type I diabetes mellitus (TIDM) development by preserving pancreatic function in NOD mice. Endocrine 2011; 40: 41-54.
  CrossrefPubMedGoogle Scholar
• 20 Barnea ER, Kirk D, Todorova K. et al. PIF direct immune regulation: Blocks mitogen- activated PBMCs proliferation, promotes T2/T1 bias, independent of Ca. Immunobiology 2015; 220: 865-875.
  CrossrefPubMedGoogle Scholar
• 21 Azar Y, Shainer R, Almogi-Hazan O. et al. Preimplantation factor reduces graftversus- host disease by regulating immune response and lowering oxidative stress (murine model). Biol Blood Marrow Transplant 2013; 19: 519-528.
  CrossrefPubMedGoogle Scholar
• 22 Mueller M, Zhou J, Yang L. et al. PreImplantation factor promotes neuroprotection by targeting microRNA let-7. Proc Natl Acad Sci USA 2014; 111: 13882-13887.
  CrossrefPubMedGoogle Scholar
• 23 Mueller M, Schoeberlein A, Zhou J. et al. PreImplantation Factor bolsters neuroprotection via modulating Protein Kinase A and Protein Kinase C signaling. Cell Death Differ 2015; 22: 2078-2086.
  CrossrefPubMedGoogle Scholar
• 24 Chen YC, Bui AV, Diesch J. et al. A novel mouse model of atherosclerotic plaque instability for drug testing and mechanistic/therapeutic discoveries using gene and microRNA expression profiling. Circ Res 2013; 113: 252-265.
  CrossrefPubMedGoogle Scholar
• 25 Greenlee-Wacker MC, Briseno C, Galvan M. et al. Membrane-associated CD93 regulates leukocyte migration and C1q-hemolytic activity during murine peritonitis. J Immunol 2011; 187: 3353-3361.
  CrossrefPubMedGoogle Scholar
• 26 Barnea ER, Lubman DM, Liu YH. et al. Insight into PreImplantation Factor (PIF*) Mechanism for Embryo Protection and Development: Target Oxidative Stress and Protein Misfolding (PDI and HSP) through Essential RIPK Binding Site. PLoS One 2014; 9: e100263.
  CrossrefPubMedGoogle Scholar
• 27 Hu L, Pennington M, Jiang Q. et al. Characterization of the functional properties of the voltage-gated potassium channel Kv1.3 in human CD4+ T lymphocytes. J Immunol 2007; 179: 4563-4570.
  CrossrefPubMedGoogle Scholar
• 28 Koenig W. Treating residual cardiovascular risk: will lipoprotein-associated phospholipase A2 inhibition live up to its promise?. J Am Coll Cardiol 2008; 51: 1642-1644.
  CrossrefPubMedGoogle Scholar
• 29 Taylor AJ, Villines TC, Stanek EJ. et al. Extended-release niacin or ezetimibe and carotid intima-media thickness. N Engl J Med 2009; 361: 2113-2122.
  CrossrefPubMedGoogle Scholar
• 30 Boden WE, Probstfield JL, Anderson T. et al. Niacin in patients with low HDL cholesterol levels receiving intensive statin therapy. N Engl J Med 2011; 365: 2255-2267.
  CrossrefPubMedGoogle Scholar
• 31 Briel M, Ferreira-Gonzalez I, You JJ. et al. Association between change in high density lipoprotein cholesterol and cardiovascular disease morbidity and mortality: systematic review and meta-regression analysis. Br Med J 2009; 338: b92.
  CrossrefPubMedGoogle Scholar
• 32 Jun M, Foote C, Lv J. et al. Effects of fibrates on cardiovascular outcomes: a systematic review and meta-analysis. Lancet 2010; 375: 1875-1884.
  CrossrefPubMedGoogle Scholar
• 33 Saha SA, Kizhakepunnur LG, Bahekar A. et al. The role of fibrates in the prevention of cardiovascular disease--a pooled meta-analysis of long-term randomized placebo-controlled clinical trials. Am Heart J 2007; 154: 943-953.
  CrossrefPubMedGoogle Scholar
• 34 Crittenden DB, Lehmann RA, Schneck L. et al. Colchicine use is associated with decreased prevalence of myocardial infarction in patients with gout. J Rheumatol 2012; 39: 1458-1464.
  CrossrefPubMedGoogle Scholar
• 35 Deftereos S, Giannopoulos G, Raisakis K. et al. Colchicine treatment for the prevention of bare-metal stent restenosis in diabetic patients. J Am Coll Cardiol 2013; 61: 1679-1685.
  CrossrefPubMedGoogle Scholar
• 36 Nidorf M, Thompson PL. Effect of colchicine (0.5 mg twice daily) on high-sensitivity C-reactive protein independent of aspirin and atorvastatin in patients with stable coronary artery disease. Am J Cardiol 2007; 99: 805-807.
  CrossrefPubMedGoogle Scholar
• 37 Nidorf SM, Eikelboom JW, Budgeon CA. et al. Low-dose colchicine for secondary prevention of cardiovascular disease. J Am Coll Cardiol 2013; 61: 404-410.
  CrossrefPubMedGoogle Scholar
• 38 Dixon WG, Watson KD, Lunt M. et al. Reduction in the incidence of myocardial infarction in patients with rheumatoid arthritis who respond to anti-tumor necrosis factor alpha therapy: results from the British Society for Rheumatology Biologics Register. Arthritis Rheum 2007; 56: 2905-2912.
  CrossrefPubMedGoogle Scholar
• 39 Georgiadis AN, Voulgari PV, Argyropoulou MI. et al. Early treatment reduces the cardiovascular risk factors in newly diagnosed rheumatoid arthritis patients. Semin Arthritis Rheum 2008; 38: 13-19.
  CrossrefPubMedGoogle Scholar
• 40 Jacobsson LT, Turesson C, Gulfe A. et al. Treatment with tumor necrosis factor blockers is associated with a lower incidence of first cardiovascular events in patients with rheumatoid arthritis. J Rheumatol 2005; 32: 1213-1218.
  PubMedGoogle Scholar
• 41 Westlake SL, Colebatch AN, Baird J. et al. The effect of methotrexate on cardiovascular disease in patients with rheumatoid arthritis: a systematic literature review. Rheumatology 2010; 49: 295-307.
  CrossrefPubMedGoogle Scholar
• 42 Reiss AB, Carsons SE, Anwar K. et al. Atheroprotective effects of methotrexate on reverse cholesterol transport proteins and foam cell transformation in human THP-1 monocyte/macrophages. Arthritis Rheum 2008; 58: 3675-3683.
  CrossrefPubMedGoogle Scholar
• 43 Ridker PM. Closing the loop on inflammation and atherothrombosis: why perform the CIRT and CANTOS trials?. Trans Am Clin Climatol Assoc 2013; 124: 174-190.
  PubMedGoogle Scholar
• 44 Voloshyna I, Littlefield MJ, Reiss AB. Atherosclerosis and interferon-gamma: new insights and therapeutic targets. Trends Cardiovasc Med 2014; 24: 45-51.
  CrossrefPubMedGoogle Scholar
• 45 Whitman SC, Ravisankar P, Daugherty A. Interleukin-18 enhances atherosclerosis in apolipoprotein E(-/-) mice through release of interferon-gamma. Circ Res 2002; 90: E34-E38.
  CrossrefPubMedGoogle Scholar
• 46 Caravaggio JW, Hasu M, MacLaren R. et al. Insulin-degrading enzyme deficiency in bone marrow cells increases atherosclerosis in LDL receptor-deficient mice. Cardiovasc Pathol 2013; 22: 458-464.
  CrossrefPubMedGoogle Scholar
• 47 Koulis C, Chen YC, Hausding C. et al. Protective role for Toll-like receptor-9 in the development of atherosclerosis in apolipoprotein E-deficient mice. Arterioscl Thromb Vasc Biol 2014; 34: 516-525.
  CrossrefPubMedGoogle Scholar
• 48 Ridker PM, Cannon CP, Morrow D. et al. C-reactive protein levels and outcomes after statin therapy. N Engl J Med 2005; 352: 20-28.
  CrossrefPubMedGoogle Scholar
• 49 Lv S, Liu Y, Zou Z. et al. The impact of statins therapy on disease activity and inflammatory factor in patients with rheumatoid arthritis: a meta-analysis. Clin Exp Rheumatol 2015; 33: 69-76.
  PubMedGoogle Scholar

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