Applied Microbiology and Biotechnology

, Volume 100, Issue 12, pp 5353–5361 | Cite as

TAT-HSA-α-MSH fusion protein with extended half-life inhibits tumor necrosis factor-α in brain inflammation of mice

  • Meizhu Wang
  • Dejuan Zhi
  • Haiqing Wang
  • Yi Ru
  • Hui Ren
  • Na Wang
  • Yiyao Liu
  • Yang Li
  • Hongyu LiEmail author
Biotechnological products and process engineering


Neuroinflammation constitutes a principal process involved in the progression of various central nervous system (CNS) disorders, including Parkinson’s disease, Alzheimer’s disease, ischemic stroke, and traumatic brain injury. The safety and efficacy of potential neuroprotective therapeutic agents is controversial and limited. Alpha-melanocyte-stimulating hormone (α-MSH) as a tridecapeptide derived from pro-opiomelanocortin displays potent anti-inflammatory and protective effects with a wide therapeutic window in brain damage. However, it is difficult to deliver effective concentrations of α-MSH into brain tissue via nondirect application. Besides, the half-life of the tridecapeptide is only a few minutes. In the present study, we generated a novel TAT-HSA-α-MSH by genetically fusing α-MSH with N-terminus 11-amino acid protein transduction domain of the human immunodeficiency virus Tat protein (TAT) and human serum albumin (HSA), which showed favorable pharmacokinetic properties and can effectively cross the blood brain barrier (BBB). The findings showed that TAT-HSA-α-MSH significantly inhibits NF-κB activation in human glioma cells A172 and tumor necrosis factor-α (TNF-α) production in experimental brain inflammation. These results indicate that TAT-HSA-α-MSH may be a potential therapeutic agent for treating neuroinflammation which plays a fundamental role in CNS disorders.


Half-life Neuroinflammation Blood brain barrier TAT-HSA-α-MSH 



This study was supported by the special international cooperation project of Ministry of Science and Technology (2012DFA30480); the key project of Gansu Province science and technology (1002WKDE055); Gansu Province Scientific Research Project Fund (090WCGA900). Fundamental Research Funds for the Central Universities of China (lzujbky-2014-147).

Compliance with ethical standards

The whole experiments involving mice were approved by the Ethics Committee of Experimental Animals of Lanzhou University and carried out in accordance with the Guide for the Care and Use of Laboratory Animals.

Conflict of interest

The authors declare that they have no competing interests.


  1. Ahmed TJ, Montero-Melendez T, Perretti M, Pitzalis C (2013) Curbing inflammation through endogenous pathways: focus on melanocortin peptides. Int J Inflamm 2013:985815. doi: 10.1155/2013/985815 CrossRefGoogle Scholar
  2. Allison DJ, Ditor DS (2014) The common inflammatory etiology of depression and cognitive impairment: a therapeutic target. J Neuroinflammation 11:151. doi: 10.1186/s12974-014-0151-1 CrossRefPubMedPubMedCentralGoogle Scholar
  3. Andersen JT, Pehrson R, Tolmachev V, Daba MB, Abrahmsen L, Ekblad C (2011) Extending half-life by indirect targeting of the neonatal Fc receptor (FcRn) using a minimal albumin binding domain. J Biol Chem 286:5234–5241. doi: 10.1074/jbc.M110.164848 CrossRefPubMedPubMedCentralGoogle Scholar
  4. Asoh S, Ohta S (2008) PTD-mediated delivery of anti-cell death proteins/peptides and therapeutic enzymes. Adv Drug Deliv Rev 60:499–516. doi: 10.1016/j.addr.2007.09.011 CrossRefPubMedGoogle Scholar
  5. Brzoska T, Luger TA, Maaser C, Abels C, Bohm M (2008) Alpha-melanocyte-stimulating hormone and related tripeptides: biochemistry, antiinflammatory and protective effects in vitro and in vivo, and future perspectives for the treatment of immune-mediated inflammatory diseases. Endocr Rev 29:581–602. doi: 10.1210/er.2007-0027 CrossRefPubMedGoogle Scholar
  6. Cai B, Lin Y, Xue XH, Fang L, Wang N, Wu ZY (2011) TAT-mediated delivery of neuroglobin protects against focal cerebral ischemia in mice. Exp Neurol 227:224–231. doi: 10.1016/j.expneurol.2010.11.009 CrossRefPubMedGoogle Scholar
  7. Catania A (2007) The melanocortin system in leukocyte biology. J Leukoc Biol 81:383–392. doi: 10.1189/jlb.0706426 CrossRefPubMedGoogle Scholar
  8. Catania A (2008) Neuroprotective actions of melanocortins: a therapeutic opportunity. Trends Neurosci 31:353–360. doi: 10.1016/j.tins.2008.04.002 CrossRefPubMedGoogle Scholar
  9. Catania A, Gatti S, Colombo G, Lipton JM (2004) Targeting melanocortin receptors as a novel strategy to control inflammation. Pharmacol Rev 56:1–29. doi: 10.1124/pr.56.1.1 CrossRefPubMedGoogle Scholar
  10. Catania A, Lonati C, Sordi A, Carlin A, Leonardi P, Gatti S (2010) The melanocortin system in control of inflammation. Sci World J 10:1840–1853. doi: 10.1100/tsw.2010.173 CrossRefGoogle Scholar
  11. Chen X, Zaro JL, Shen WC (2013) Fusion protein linkers: property, design and functionality. Adv Drug Deliv Rev 65:1357–1369. doi: 10.1016/j.addr.2012.09.039 CrossRefPubMedPubMedCentralGoogle Scholar
  12. Chuang VTG, Kragh-Hansen U, Otagiri M (2002) Pharmaceutical strategies utilizing recombinant human serum albumin. Pharm Res-Dordr 19:569–576CrossRefGoogle Scholar
  13. Doeppner TR, El Aanbouri M, Dietz GP, Weise J, Schwarting S, Bahr M (2010) Transplantation of TAT-Bcl-xL-transduced neural precursor cells: long-term neuroprotection after stroke. Neurobiol Dis 40:265–276. doi: 10.1016/j.nbd.2010.05.033 CrossRefPubMedGoogle Scholar
  14. Duttaroy A, Kanakaraj P, Osborn BL, Schneider H, Pickeral OK, Chen C, Zhang G, Kaithamana S, Singh M, Schulingkamp R, Crossan D, Bock J, Kaufman TE, Reavey P, Barber MC, Krishnan SR, Garcia A, Murphy K, Siskind JK, McLean MA, Cheng S, Ruben S, Birse CE, Blonde O (2005) Development of a Long-Acting Insulin Analog Using Albumin Fusion Technology. Diabetes 54:251–258Google Scholar
  15. Fanali G, di Masi A, Trezza V, Marino M, Fasano M, Ascenzi P (2012) Human serum albumin: from bench to bedside. Mol Asp Med 33:209–290. doi: 10.1016/j.mam.2011.12.002 CrossRefGoogle Scholar
  16. Finsen B, Owens T (2011) Innate immune responses in central nervous system inflammation. FEBS Lett 585:3806–3812. doi: 10.1016/j.febslet.2011.05.030 CrossRefPubMedGoogle Scholar
  17. Forslin Aronsson A, Spulber S, Oprica M, Winblad B, Post C, Schultzberg M (2007) Alpha-MSH rescues neurons from excitotoxic cell death. J Mol Neurosci:MN 33:239–251. doi: 10.1007/s12031-007-0019-2 CrossRefPubMedGoogle Scholar
  18. Gantz I, Fong TM (2003) The melanocortin system. Am J Physiol Endocrinol Metab 284:E468–E474. doi: 10.1152/ajpendo.00434.2002 CrossRefPubMedGoogle Scholar
  19. Giuliani D, Minutoli L, Ottani A, Spaccapelo L, Bitto A, Galantucci M, Altavilla D, Squadrito F, Guarini S (2012) Melanocortins as potential therapeutic agents in severe hypoxic conditions. Front Neuroendocrinol 33:179–193. doi: 10.1016/j.yfrne.2012.04.001 CrossRefPubMedGoogle Scholar
  20. Gonzalez H, Elgueta D, Montoya A, Pacheco R (2014) Neuroimmune regulation of microglial activity involved in neuroinflammation and neurodegenerative diseases. J Neuroimmunol 274:1–13. doi: 10.1016/j.jneuroim.2014.07.012 CrossRefPubMedGoogle Scholar
  21. Gump JM, Dowdy SF (2007) TAT transduction: the molecular mechanism and therapeutic prospects. Trends Mol Med 13:443–448. doi: 10.1016/j.molmed.2007.08.002 CrossRefPubMedGoogle Scholar
  22. Holdeman M, Lipton JM (1985) Antipyretic activity of a potent α-MSH Analog. Peptides 6:273–275CrossRefPubMedGoogle Scholar
  23. Huang Y, Rao Y, Feng C, Li Y, Wu X, Su Z, Xiao J, Xiao Y, Feng W, Li X (2008) High-level expression and purification of Tat-haFGF19-154. Appl Microbiol Biotechnol 77:1015–1022. doi: 10.1007/s00253-007-1249-5 CrossRefPubMedGoogle Scholar
  24. Ichiyama T, Zhao H, Catania A, Furukawa S, Lipton JM (1999) alpha-melanocyte-stimulating hormone inhibits NF-kappaB activation and IkappaBalpha degradation in human glioma cells and in experimental brain inflammation. Exp Neurol 157:359–365CrossRefPubMedGoogle Scholar
  25. Jeong HJ, Kim DW, Kim MJ, Woo SJ, Kim HR, Kim SM, Jo HS, Hwang HS, Kim DS, Cho SW, Won MH, Han KH, Park JS, Eum WS, Choi SY (2012) Protective effects of transduced Tat-DJ-1 protein against oxidative stress and ischemic brain injury. Exp Mol Med 44:586–593. doi: 10.3858/emm.2012.44.10.067 CrossRefPubMedPubMedCentralGoogle Scholar
  26. Kielian T (2006) Toll-like receptors in central nervous system glial inflammation and homeostasis. J Neurosci Res 83:711–730. doi: 10.1002/jnr.20767 CrossRefPubMedPubMedCentralGoogle Scholar
  27. Kincses ZT, Vecsei L (2011) Pharmacological therapy in Parkinson’s disease: focus on neuroprotection. CNS Neurosci Ther 17:345–367. doi: 10.1111/j.1755-5949.2010.00150.x CrossRefPubMedGoogle Scholar
  28. Koistinaho M, Koistinaho J (2005) Interactions between Alzheimer’s disease and cerebral ischemia—focus on inflammation. Brain Res Rev 48:240–250. doi: 10.1016/j.brainresrev.2004.12.014 CrossRefPubMedGoogle Scholar
  29. Kratz F (2008) Albumin as a drug carrier: design of prodrugs, drug conjugates and nanoparticles. J Control Release 132:171–183. doi: 10.1016/j.jconrel.2008.05.010 CrossRefPubMedGoogle Scholar
  30. Kumar A, Loane DJ (2012) Neuroinflammation after traumatic brain injury: opportunities for therapeutic intervention. Brain Behav Immun 26:1191–1201. doi: 10.1016/j.bbi.2012.06.008 CrossRefPubMedGoogle Scholar
  31. Lucas SM, Rothwell NJ, Gibson RM (2006) The role of inflammation in CNS injury and disease. Br J Pharmacol 147(Suppl 1):S232–S240. doi: 10.1038/sj.bjp.0706400 PubMedPubMedCentralGoogle Scholar
  32. Mattson MP, Camandola S (2001) NF-κB in neuronal plasticity and neurodegenerative disorders. J Clin Invest 107(3):247–254CrossRefPubMedPubMedCentralGoogle Scholar
  33. Merkler DJ (1994) C-Terminal amidated peptides: production by the in vitro enzymatic amidation of glycine-extended peptides and the importance of the amide to bioactivity. Enzyme Microb Tech. 116Google Scholar
  34. Miller AH, Maletic V, Raison CL (2009) Inflammation and its discontents: the role of cytokines in the pathophysiology of major depression. Biol Psychiatry 65:732–741. doi: 10.1016/j.biopsych.2008.11.029 CrossRefPubMedPubMedCentralGoogle Scholar
  35. Minnerup J, Sutherland BA, Buchan AM, Kleinschnitz C (2012) Neuroprotection for stroke: current status and future perspectives. Int J Mol Sci 13:11753–11772. doi: 10.3390/ijms130911753 CrossRefPubMedPubMedCentralGoogle Scholar
  36. Osborn BL, Sekut L, Corcoran M, Poortman C, Sturm B, Chen G, Mather D, Lin HL, Parry TJ (2002) Albutropin: a growth hormone–albumin fusion with improved pharmacokinetics and pharmacodynamics in rats and monkeys. Eur J Pharmacol 456:149–158Google Scholar
  37. Paxinos G, Franklin KBJ (2002) The mouse brain in stereotaxic coordinates. Academic, New YorkGoogle Scholar
  38. Prajapati KD, Sharma SS, Roy N (2011) Current perspectives on potential role of albumin in neuroprotection. Rev Neurosci 22:355–363. doi: 10.1515/RNS.2011.028 CrossRefPubMedGoogle Scholar
  39. Rajora N, Boccoli G, Burns D, Sharma S, Catania AP, Lipton JM (1997) alpha-MSH modulates local and circulating tumor necrosis factor-alpha in experimental brain. J Neurosci 17(6):2181–2186PubMedGoogle Scholar
  40. Schwarze SR, Ho A, Vocero-Akbani A, Dowdy SF (1999) In vivo protein transduction: delivery of a biologically active protein into the mouse. Science 285:1569–1572CrossRefPubMedGoogle Scholar
  41. Subramanian GM, Fiscella M, Lamouse-Smith A, Zeuzem S, McHutchison JG (2007) Albinterferon alpha-2b: a genetic fusion protein for the treatment of chronic hepatitis C. Nat Biotechnol 25:1411–1419. doi: 10.1038/nbt1364
  42. Tanaka R, Ishima Y, Enoki Y, Kimachi K, Shirai T, Watanabe H, Chuang VT, Maruyama T, Otagiri (2014) Therapeutic impact of human serum albumin-thioredoxin fusion protein on influenza virus-induced lung injury mice. Front Immunol 5:561. doi: 10.3389/fimmu.2014.00561 CrossRefPubMedPubMedCentralGoogle Scholar
  43. Thompson AM, Trujillo JM (2015) Dulaglutide: the newest GLP-1 receptor agonist for the management of type 2 diabetes. Ann Pharmacother 49:351–359. doi: 10.1177/1060028014564180 CrossRefPubMedGoogle Scholar
  44. Tsubaki M, Terashima I, Kamata K, Koga A (2013) C-terminal modification of monoclonal antibody drugs: amidated species as a general product-related substance. Int J Biol Macromol 52:139–147. doi: 10.1016/j.ijbiomac.2012.09.016
  45. Tyagi E, Agrawal R, Nath C, Shukla R (2008) Influence of LPS-induced neuroinflammation on acetylcholinesterase activity in rat brain. J Neuroimmunol 205:51–56. doi: 10.1016/j.jneuroim.2008.08.015 CrossRefPubMedGoogle Scholar
  46. Wilson JF (1988) Low permeability of the blood-brain barrier to nanomolar concentrations of immunoreactive alpha-melanotropin. Psychopharmacology 96:262–266CrossRefPubMedGoogle Scholar
  47. Yin P, Luby TM, Chen H, Etemad-Moghadam B, Lee D, Aziz N, Ramstedt U, Hedley ML (2003) Generation of expression constructs that secrete bioactive alphaMSH and their use in the treatment of experimental autoimmune encephalomyelitis. Gene Ther 10:348–355. doi: 10.1038/ CrossRefPubMedGoogle Scholar
  48. Zhu Y, Bu Q, Liu X, Hu W, Wang Y (2014) Neuroprotective effect of TAT-14-3-3epsilon fusion protein against cerebral ischemia/reperfusion injury in rats. PLoS One 9:e93334. doi: 10.1371/journal.pone.0093334 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Meizhu Wang
    • 1
  • Dejuan Zhi
    • 2
  • Haiqing Wang
    • 2
  • Yi Ru
    • 1
  • Hui Ren
    • 2
  • Na Wang
    • 2
  • Yiyao Liu
    • 1
  • Yang Li
    • 2
  • Hongyu Li
    • 1
    • 2
    Email author
  1. 1.Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, Institute of Microbiology, School of Life SciencesLanzhou UniversityLanzhouPeople’s Republic of China
  2. 2.Gansu High Throughput Screening and Creation Center for Health Products, School of PharmacyLanzhou UniversityLanzhouPeople’s Republic of China

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