AAPS PharmSciTech

, 20:50 | Cite as

IGF-II-Conjugated Nanocarrier for Brain-Targeted Delivery of p11 Gene for Depression

  • Manit Gandhi
  • Priyanka Bhatt
  • Gaurav Chauhan
  • Sarita Gupta
  • Ambikanandan Misra
  • Rajashree MashruEmail author
Research Article Theme: Translational Multi-Disciplinary Approach for the Drug and Gene Delivery Systems
Part of the following topical collections:
  1. Theme: Translational Multi-Disciplinary Approach for the Drug and Gene Delivery Systems


Gene therapy involving p11 cDNA has been thought to be a futuristic approach for the effective management of depression as the existing treatment regimen presents many issues regarding late onset of action, patient withdrawal and their side effects. For the effective transfection of p11 gene intracellularly, two cationic lipids based on phospholipid DOPE conjugated to basic amino acids histidine and arginine were synthesised, used for liposome formulation and evaluated for their ability as gene delivery vectors. They were further converted using IGF-II mAb into immunoliposomes for CNS targeting and mAb conjugation to liposomes were characterised by SDS-PAGE. They were further analysed by in vitro characterisation studies that include erythrocyte aggregation study, electrolyte-induced study, heparin compatibility study and serum stability studies. SHSY5Y cells were used for conducting cytotoxicity of synthesised lipids and live imaging of cell uptake for 25 min. Finally, the brain distribution studies and western blot were carried out in animals to evaluate them for their BBB permeation ability and effects on p11 protein which is believed to be a culprit. These formulated liposomes from synthesised lipids offer a promising approach for the treatment of depression.


gene therapy depression CNS targeting liposomes p11 gene 



The authors would like to thank the Department of Chemistry and Biochemistry, The Maharaja Sayajirao University of Baroda, Vadodara, for the help in carrying out lipid synthesis and plasmid isolation respectively.

Compliance with Ethical Standards

The protocol for the study was duly approved by Institutional Animal Ethics Committee, Faculty of Pharmacy, The M. S. University of Baroda, Vadodara, vide protocol approval no: MSU/IAEC/2016-17/1631.

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

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Supplementary file 1

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Supplementary file 2

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High resolution image (TIF 113 kb)
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Supplementary file 3 (PNG 62 kb)


  1. 1.
    Svenningsson P, Kim Y, Warner-Schmidt J, Oh Y-S, Greengard P. p11 and its role in depression and therapeutic responses to antidepressants. Nat Rev Neurosci. 2013;14(10):673–80.CrossRefGoogle Scholar
  2. 2.
    Warner-Schmidt JL, Vanover KE, Chen EY, Marshall JJ, Greengard P. Antidepressant effects of selective serotonin reuptake inhibitors (SSRIs) are attenuated by antiinflammatory drugs in mice and humans. Proc Natl Acad Sci. 2011;108(22):9262–7.CrossRefGoogle Scholar
  3. 3.
    Lalani R, Misra A, Amrutiya J, Patel H, Bhatt P, Patil SK. Approaches and recent trends in gene delivery for treatment of atherosclerosis. Recent Pat Drug Deliv Formul. 2016;10(2):141–55.CrossRefGoogle Scholar
  4. 4.
    Lalani RA, Bhatt P, Rathi M, Misra A. Abstract 2063: improved sensitivity and in vitro efficacy of RGD grafted PEGylated gemcitabine liposomes in RRM1 siRNA pretreated cancer cells. Cancer Research. 2016;76(14 Supplement):2063. Scholar
  5. 5.
    Rehman Z, Zuhorn IS, Hoekstra D. How cationic lipids transfer nucleic acids into cells and across cellular membranes: recent advances. J Control Release. 2013;166(1):46–56.CrossRefGoogle Scholar
  6. 6.
    Fitzsimmons R. Novel polymer and lipid-based nanocarriers for gene delivery. 2011.Google Scholar
  7. 7.
    Tsuji A, Tamai I. Blood-brain barrier function of P-glycoprotein. Adv Drug Deliv Rev. 1997;25(2–3):287–98.CrossRefGoogle Scholar
  8. 8.
    Bremner JB, Samosorn S, Ambrus JI. N-Acylation of 5-substituted indoles with carboxylic acids via DCC coupling. Synthesis. 2004;2004(16):2653–8.CrossRefGoogle Scholar
  9. 9.
    Vhora I, Lalani R, Bhatt P, Patil S, Patel H, Patel V, et al. Colloidally stable small unilamellar stearyl amine lipoplexes for effective BMP-9 gene delivery to stem cells for osteogenic differentiation. AAPS PharmSciTech. 2018.
  10. 10.
    Sasaki Y, Coy DH. Solid phase synthesis of peptides containing the CH2NH peptide bond isostere. Peptides. 1987;8(1):119–21.CrossRefGoogle Scholar
  11. 11.
    Patil S, Bhatt P, Lalani R, Amrutiya J, Vhora I, Kolte A, et al. Low molecular weight chitosan–protamine conjugate for siRNA delivery with enhanced stability and transfection efficiency. RSC Adv. 2016;6(112):110951–63. Scholar
  12. 12.
    Zhang Q-F, Yang W-H, Yi W-J, Zhang J, Ren J, Luo T-Y, et al. TACN-containing cationic lipids with ester bond: preparation and application in gene delivery. Bioorg Med Chem Lett. 2011;21(23):7045–9.CrossRefGoogle Scholar
  13. 13.
    Bhatt P, Lalani R, Vhora I, Patil S, Amrutiya J, Misra A, et al. Liposomes encapsulating native and cyclodextrin enclosed paclitaxel: enhanced loading efficiency and its pharmacokinetic evaluation. Int J Pharm. 2018;536(1):95–107. Scholar
  14. 14.
    Zensi A, Begley D, Pontikis C, Legros C, Mihoreanu L, Wagner S, et al. Albumin nanoparticles targeted with Apo E enter the CNS by transcytosis and are delivered to neurones. J Control Release. 2009;137(1):78–86.CrossRefGoogle Scholar
  15. 15.
    Bhatt P, Lalani R, Mashru R, Misra A. Abstract 2065: anti-FSHR antibody Fab’ fragment conjugated immunoliposomes loaded with cyclodextrin-paclitaxel complex for improved <em>in vitro</em> efficacy on ovarian cancer cells. Cancer Res. 2016;76(14 Supplement):2065. Scholar
  16. 16.
    Perrier T, Saulnier P, Benoît JP. Methods for the functionalisation of nanoparticles: new insights and perspectives. Chem Eur J. 2010;16(38):11516–29.CrossRefGoogle Scholar
  17. 17.
    Yewale C, Baradia D, Patil S, Bhatt P, Amrutiya J, Gandhi R, et al. Docetaxel loaded immunonanoparticles delivery in EGFR overexpressed breast carcinoma cells. Journal of Drug Delivery Science and Technology. 2018;45:334–45. Scholar
  18. 18.
    Sölch S. Effects of the GTPase Rab31 on breast cancer cell proliferation, adhesion, and expression of other tumor-associated genes. Munich: Technische Universität München; 2014.Google Scholar
  19. 19.
    Sasse J, Gallagher SR. Staining proteins in gels. Curr Protoc Mol Biol. 2009;10:6. 1–6. 27.Google Scholar
  20. 20.
    Stebelska K, Dubielecka PM, Sikorski AF. The effect of PS content on the ability of natural membranes to fuse with positively charged liposomes and lipoplexes. J Membr Biol. 2005;206(3):203–14.CrossRefGoogle Scholar
  21. 21.
    Lee S-Y, Huh MS, Lee S, Lee SJ, Chung H, Park JH, et al. Stability and cellular uptake of polymerized siRNA (poly-siRNA)/polyethylenimine (PEI) complexes for efficient gene silencing. J Control Release. 2010;141(3):339–46.CrossRefGoogle Scholar
  22. 22.
    Bhatt P, Khatri N, Kumar M, Baradia D, Misra A. Microbeads mediated oral plasmid DNA delivery using polymethacrylate vectors: an effectual groundwork for colorectal cancer. Drug Deliv. 2015;22(6):849–61. Scholar
  23. 23.
    Nguyen LT, Atobe K, Barichello JM, Ishida T, Kiwada H. Complex formation with plasmid DNA increases the cytotoxicity of cationic liposomes. Biol Pharm Bull. 2007;30(4):751–7.CrossRefGoogle Scholar
  24. 24.
    Katsarou K, Lavdas AΑ, Tsitoura P, Serti E, Markoulatos P, Mavromara P, et al. Endocytosis of hepatitis C virus non-enveloped capsid-like particles induces MAPK–ERK1/2 signaling events. Cell Mol Life Sci. 2010;67(14):2491–506.CrossRefGoogle Scholar
  25. 25.
    MacKay JA, Deen DF, Szoka FC Jr. Distribution in brain of liposomes after convection enhanced delivery; modulation by particle charge, particle diameter, and presence of steric coating. Brain Res. 2005;1035(2):139–53.CrossRefGoogle Scholar
  26. 26.
    Walsh DM, Klyubin I, Fadeeva JV, Cullen WK, Anwyl R, Wolfe MS, et al. Naturally secreted oligomers of amyloid β protein potently inhibit hippocampal long-term potentiation in vivo. Nature. 2002;416(6880):535–9.CrossRefGoogle Scholar
  27. 27.
    Poceva Panovska A, Acevska J, Stefkov G, Brezovska K, Petkovska R, Dimitrovska A. Optimization of HS-GC–FID–MS method for residual solvent profiling in active pharmaceutical ingredients using DoE. J Chromatogr Sci. 2015;54(2):103–11.PubMedGoogle Scholar
  28. 28.
    Dhillon S, Von Burg R. Methylene chloride. J Appl Toxicol. 1995;15(4):329–35.CrossRefGoogle Scholar
  29. 29.
    Brown M, Schätzlein A, Brownlie A, Jack V, Wang W, Tetley L, et al. Preliminary characterization of novel amino acid based polymeric vesicles as gene and drug delivery agents. Bioconjug Chem. 2000;11(6):880–91.CrossRefGoogle Scholar
  30. 30.
    Pouton CW, Seymour LW. Key issues in non-viral gene delivery1. Adv Drug Deliv Rev. 2001;46(1–3):187–203.CrossRefGoogle Scholar
  31. 31.
    Bhatt P, Vhora I, Patil S, Amrutiya J, Bhattacharya C, Misra A, et al. Role of antibodies in diagnosis and treatment of ovarian cancer: basic approach and clinical status. J Control Release. 2016;226:148–67. Scholar
  32. 32.
    Koren E, Apte A, Jani A, Torchilin VP. Multifunctional PEGylated 2C5-immunoliposomes containing pH-sensitive bonds and TAT peptide for enhanced tumor cell internalization and cytotoxicity. J Control Release. 2012;160(2):264–73.CrossRefGoogle Scholar
  33. 33.
    Knežević-Jugović ZD, Bezbradica DI, Mijin DŽ, Antov MG. The immobilization of enzyme on Eupergit® supports by covalent attachment. Enzyme Stabilization and Immobilization. Berlin: Springer; 2011. p. 99–111.CrossRefGoogle Scholar
  34. 34.
    Tandel H, Bhatt P, Jain K, Shahiwala A, Misra A. In-vitro and in-vivo tools in emerging drug delivery scenario: challenges and updates. In: Misra ASA, editor. In-vitro and in-vivo tools in drug delivery research for optimum clinical outcomes. Boca Raton: CRC Press; 2018.Google Scholar
  35. 35.
    Asnis GM, De La Garza R. Interferon-induced depression in chronic hepatitis C: a review of its prevalence, risk factors, biology, and treatment approaches. J Clin Gastroenterol. 2006;40(4):322–35.CrossRefGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2018

Authors and Affiliations

  • Manit Gandhi
    • 1
  • Priyanka Bhatt
    • 1
  • Gaurav Chauhan
    • 1
  • Sarita Gupta
    • 1
  • Ambikanandan Misra
    • 1
  • Rajashree Mashru
    • 1
    Email author
  1. 1.Pharmacy Department, Faculty of Technology and EngineeringThe Maharaja Sayajirao University of BarodaVadodaraIndia

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