We’re sorry, something doesn't seem to be working properly.

Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.


Polylactide-Based Reactive Micelles as a Robust Platform for mRNA Delivery

  • 121 Accesses



mRNA has recently emerged as a potent therapeutics and requires safe and effective delivery carriers, particularly prone to address its issues of poor stability and escape from endosomes. In this context, we designed poly(D,L-lactide) (PLA)-based micelles with N-succinimidyl (NS) ester decorated hydrophilic hairy corona to trap/couple a cationic fusogenic peptide and further complex mRNA.


Two strategies were investigated, namely (i) sequential immobilization of peptide and mRNA onto the micelles (layer-by-layer, LbL) or (ii) direct immobilization of peptide-mRNA pre-complex (PC) on the micelles. After characterization by means of size, surface charge, peptide/mRNA coupling/complexation and mRNA serum stability, carrier cytotoxicity and transfection capacity were evaluated with dendritic cells (DCs) using both GFP and luciferase mRNAs.


Whatever the approach used, the micellar assemblies afforded full protection of mRNA in serum while the peptide-mRNA complex yielded complete mRNA degradation. In addition, the micellar assemblies allowed to significantly reduce the toxicity observed with the peptide-mRNA complex. They successfully transfected hard-to transfect DCs, with a superior efficiency for the LbL made ones (whatever mRNAs studied) showing the impact of the elaboration process on the carrier properties.


These results show the relevance and potential of this new PLA/peptide based micelle platform to improve mRNA stability and delivery, while offering the possibility of further multifunctionality through PLA core encapsulation.

This is a preview of subscription content, log in to check access.

Access options

Buy single article

Instant unlimited access to the full article PDF.

US$ 39.95

Price includes VAT for USA

Subscribe to journal

Immediate online access to all issues from 2019. Subscription will auto renew annually.

US$ 199

This is the net price. Taxes to be calculated in checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7


  1. 1.

    Burnett JC, Rossi JJ. RNA-based therapeutics: current Progress and future prospects. Chem Biol. 2012;19:60–71.

  2. 2.

    Kaczmarek JC, Kowalski PS, Anderson DG. Advances in the delivery of RNA therapeutics: from concept to clinical reality. Genome Med [Internet]. 2017 [cited 2018 Jul 3];9. Available from: http://genomemedicine.biomedcentral.com/articles/10.1186/s13073-017-0450-0

  3. 3.

    Subhan MA, Torchilin VP. Efficient nanocarriers of siRNA therapeutics for cancer treatment. Transl Res [Internet]. 2019 [cited 2019 Sep 3]; Available from: https://linkinghub.elsevier.com/retrieve/pii/S1931524419301392

  4. 4.

    Resnier P, Montier T, Mathieu V, Benoit J-P, Passirani C. A review of the current status of siRNA nanomedicines in the treatment of cancer. Biomaterials. 2013;34:6429–43.

  5. 5.

    Kang K-N, Lee Y-S. RNA Aptamers: A Review of Recent Trends and Applications. In: Zhong J-J, editor. Future Trends Biotechnol [Internet]. Berlin, Heidelberg: Springer Berlin Heidelberg; 2012 [cited 2018 Jul 3]. p. 153–69. Available from: http://link.springer.com/10.1007/10_2012_136

  6. 6.

    Midoux P, Pichon C. Lipid-based mRNA vaccine delivery systems. Expert Rev Vaccines. 2015;14:221–34.

  7. 7.

    Pardi N, Hogan MJ, Pelc RS, Muramatsu H, Andersen H, DeMaso CR, et al. Zika virus protection by a single low-dose nucleoside-modified mRNA vaccination. Nature. 2017;543:248–51.

  8. 8.

    Linares-Fernández S, Lacroix C, Exposito J-Y, Verrier B. Tailoring mRNA Vaccine to Balance Innate/Adaptive Immune Response. Trends Mol Med [Internet]. 2019 [cited 2019 Dec 2]; Available from: https://linkinghub.elsevier.com/retrieve/pii/S1471491419302448

  9. 9.

    Liu. A Comparison of Plasmid DNA and mRNA as Vaccine Technologies. Vaccines. 2019:7–37.

  10. 10.

    Geall AJ, Verma A, Otten GR, Shaw CA, Hekele A, Banerjee K, et al. Nonviral delivery of self-amplifying RNA vaccines. Proc Natl Acad Sci. 2012;109:14604–9.

  11. 11.

    Kranz LM, Diken M, Haas H, Kreiter S, Loquai C, Reuter KC, et al. Systemic RNA delivery to dendritic cells exploits antiviral defence for cancer immunotherapy. Nature. 2016;534:396–401.

  12. 12.

    Pollard C, Rejman J, De Haes W, Verrier B, Van Gulck E, Naessens T, et al. Type I IFN counteracts the induction of antigen-specific immune responses by lipid-based delivery of mRNA vaccines. Mol Ther. 2013;21:251–9.

  13. 13.

    Sayour EJ, De Leon G, Pham C, Grippin A, Kemeny H, Chua J, et al. Systemic activation of antigen-presenting cells via RNA-loaded nanoparticles. OncoImmunology. 2017;6:e1256527.

  14. 14.

    Jayaraman M, Ansell SM, Mui BL, Tam YK, Chen J, Du X, et al. Maximizing the potency of siRNA lipid nanoparticles for hepatic gene silencing in vivo. Angew Chem Int Ed. 2012;51:8529–33.

  15. 15.

    Pardi N, Secreto AJ, Shan X, Debonera F, Glover J, Yi Y, et al. Administration of nucleoside-modified mRNA encoding broadly neutralizing antibody protects humanized mice from HIV-1 challenge. Nat Commun. 2017;8:14630.

  16. 16.

    Richner JM, Himansu S, Dowd KA, Butler SL, Salazar V, Fox JM, et al. Modified mRNA Vaccines Protect against Zika Virus Infection. Cell. 2017;168:1114–1125.e10.

  17. 17.

    Stadler CR, Bähr-Mahmud H, Celik L, Hebich B, Roth AS, Roth RP, et al. Elimination of large tumors in mice by mRNA-encoded bispecific antibodies. Nat Med. 2017;23:815–7.

  18. 18.

    Zhao M, Li M, Zhang Z, Gong T, Sun X. Induction of HIV-1 gag specific immune responses by cationic micelles mediated delivery of gag mRNA. Drug Deliv 2015;1–12.

  19. 19.

    Wang LL, Sloand JN, Gaffey AC, Venkataraman CM, Wang Z, Trubelja A, et al. Injectable, guest–host assembled Polyethylenimine hydrogel for siRNA delivery. Biomacromolecules. 2017;18:77–86.

  20. 20.

    Chahal JS, Khan OF, Cooper CL, McPartlan JS, Tsosie JK, Tilley LD, et al. Dendrimer-RNA nanoparticles generate protective immunity against lethal Ebola, H1N1 influenza, and Toxoplasma gondii challenges with a single dose. Proc Natl Acad Sci. 2016;113:E4133–42.

  21. 21.

    Su X, Fricke J, Kavanagh DG, Irvine DJ. In Vitro and in Vivo mRNA delivery using lipid-enveloped pH-responsive polymer nanoparticles. Mol Pharm. 2011;8:774–87.

  22. 22.

    Yan Y, Xiong H, Zhang X, Cheng Q, Siegwart DJ. Systemic mRNA delivery to the lungs by functional polyester-based carriers. Biomacromolecules. 2017;18:4307–15.

  23. 23.

    Kichler A, Leborgne C, März J, Danos O, Bechinger B. Histidine-rich amphipathic peptide antibiotics promote efficient delivery of DNA into mammalian cells. Proc Natl Acad Sci. 2003;100:1564–8.

  24. 24.

    Lam JKW, Liang W, Lan Y, Chaudhuri P, Chow MYT, Witt K, et al. Effective endogenous gene silencing mediated by pH responsive peptides proceeds via multiple pathways. J Control Release. 2012;158:293–303.

  25. 25.

    McCarthy HO, McCaffrey J, McCrudden CM, Zholobenko A, Ali AA, McBride JW, et al. Development and characterization of self-assembling nanoparticles using a bio-inspired amphipathic peptide for gene delivery. J Control Release. 2014;189:141–9.

  26. 26.

    Borguet YP, Khan S, Noel A, Gunsten SP, Brody SL, Elsabahy M, et al. Development of fully degradable Phosphonium-functionalized Amphiphilic Diblock copolymers for nucleic acids delivery. Biomacromolecules. 2018;19:1212–22.

  27. 27.

    Nouri FS, Wang X, Dorrani M, Karjoo Z, Hatefi A. A recombinant biopolymeric platform for reliable evaluation of the activity of pH-responsive Amphiphile Fusogenic peptides. Biomacromolecules. 2013;14:2033–40.

  28. 28.

    Multicenter Phase II. Clinical trial of Genexol-PM® with gemcitabine in advanced biliary tract Cancer. Anticancer Res. 2017;37:1467–74.

  29. 29.

    Ando S, Putnam D, Pack DW, Langer R. PLGA microspheres containing plasmid DNA: preservation of supercoiled DNA via cryopreparation and carbohydrate stabilization. J Pharm Sci. 1999;88:126–30.

  30. 30.

    Walter E, Moelling K, Pavlovic J, Merkle HP. Microencapsulation of DNA using poly(dl-lactide-co-glycolide): stability issues and release characteristics. J Control Release. 1999;61:361–74.

  31. 31.

    Cun D, Foged C, Yang M, Frøkjær S, Nielsen HM. Preparation and characterization of poly(dl-lactide-co-glycolide) nanoparticles for siRNA delivery. Int J Pharm. 2010;390:70–5.

  32. 32.

    Jain AK, Massey A, Yusuf H, Kett VL, McDonald D, McCarthy H. Development of polymeric–cationic peptide composite nanoparticles, a nanoparticle-in-nanoparticle system for controlled gene delivery. Int J Nanomedicine. 2015;7183.

  33. 33.

    Tinsley-Bown AM, Fretwell R, Dowsett AB, Davis SL, Farrar GH. Formulation of poly(d,l-lactic-co-glycolic acid) microparticles for rapid plasmid DNA delivery. J Control Release. 2000;66:229–41.

  34. 34.

    O’Hagan D, Singh M, Ugozzoli M, Wild C, Barnett S, Chen M, et al. Induction of potent immune responses by cationic microparticles with adsorbed human immunodeficiency virus DNA vaccines. J Virol. 2001;75:9037–43.

  35. 35.

    Maruyama A, Ishihara T, Kim J-S, Kim SW, Akaike T. Nanoparticle DNA Carrier with Poly( l -lysine) Grafted Polysaccharide Copolymer and Poly( d , l -lactic acid). Bioconjug Chem. 1997;8:735–42 Nanoparticle DNA carrier with poly(L-lysine) grafted polysaccharide copolymer and poly(D,L-lactic acid).

  36. 36.

    Trimaille T, Pichot C, Delair T. Surface functionalization of poly(d,l-lactic acid) nanoparticles with poly(ethylenimine) and plasmid DNA by the layer-by-layer approach. Colloids Surf Physicochem Eng Asp. 2003;221:39–48.

  37. 37.

    Chen C-K, Jones CH, Mistriotis P, Yu Y, Ma X, Ravikrishnan A, et al. Poly(ethylene glycol)-block-cationic polylactide nanocomplexes of differing charge density for gene delivery. Biomaterials. 2013;34:9688–99.

  38. 38.

    Ni Q, Zhang F, Zhang Y, Zhu G, Wang Z, Teng Z, et al. In situ shRNA synthesis on DNA-Polylactide nanoparticles to treat multidrug resistant breast Cancer. Adv Mater. 2018;30:1705737.

  39. 39.

    Handké N, Lahaye V, Bertin D, Delair T, Verrier B, Gigmes D, et al. Elaboration of glycopolymer-functionalized micelles from an N-vinylpyrrolidone/lactide-based reactive copolymer platform. Macromol Biosci. 2013;13:1213–20.

  40. 40.

    Handké N, Trimaille T, Luciani E, Rollet M, Delair T, Verrier B, et al. Elaboration of densely functionalized polylactide nanoparticles from N-acryloxysuccinimide-based block copolymers. J Polym Sci Part Polym Chem. 2011;49:1341–50.

  41. 41.

    Jiménez-Sánchez G, Terrat C, Verrier B, Gigmes D, Trimaille T. Improving bioassay sensitivity through immobilization of bio-probes onto reactive micelles. Chem Commun. 2017;53:8062–5.

  42. 42.

    Jiménez-Sánchez G, Pavot V, Chane-Haong C, Handké N, Terrat C, Gigmes D, et al. Preparation and in vitro evaluation of Imiquimod loaded Polylactide-based micelles as potential vaccine adjuvants. Pharm Res. 2015;32:311–20.

  43. 43.

    Nazari M, Zamani Koukhaloo S, Mousavi S, Minai-Tehrani A, Emamzadeh R, Cheraghi R. Development of a ZHER3-Affibody-targeted Nano-vector for gene delivery to HER3-overexpressed breast Cancer cells. Macromol Biosci. 2019;19:1900159.

  44. 44.

    Handké N, Ficheux D, Rollet M, Delair T, Mabrouk K, Bertin D, et al. Lysine-tagged peptide coupling onto polylactide nanoparticles coated with activated ester-based amphiphilic copolymer: a route to highly peptide-functionalized biodegradable carriers. Colloids Surf B Biointerfaces. 2013;103:298–303.

  45. 45.

    Bennett R, Yakkundi A, McKeen HD, McClements L, McKeogh TJ, McCrudden CM, et al. RALA-mediated delivery of FKBPL nucleic acid therapeutics. Nanomed. 2015;10:2989–3001.

  46. 46.

    Ali AA, McCrudden CM, McCaffrey J, McBride JW, Cole G, Dunne NJ, et al. DNA vaccination for cervical cancer; a novel technology platform of RALA mediated gene delivery via polymeric microneedles. Nanomedicine Nanotechnol Biol Med. 2017;13:921–32.

  47. 47.

    Udhayakumar VK, De Beuckelaer A, McCaffrey J, McCrudden CM, Kirschman JL, Vanover D, et al. Arginine-rich peptide-based mRNA Nanocomplexes efficiently instigate cytotoxic T cell immunity dependent on the amphipathic Organization of the Peptide. Adv Healthc Mater. 2017;6:1601412.

  48. 48.

    Koh KJ, Liu Y, Lim SH, Loh XJ, Kang L, Lim CY, et al. Formulation, characterization and evaluation of mRNA-loaded dissolvable polymeric microneedles (RNApatch). Sci Rep [Internet]. 2018 [cited 2019 Aug 16];8. Available from: http://www.nature.com/articles/s41598-018-30290-3

  49. 49.

    McLenachan S, Zhang D, Palomo ABA, Edel MJ, Chen FK. mRNA Transfection of Mouse and Human Neural Stem Cell Cultures. Wu Q, editor. PLoS ONE. 2013;8:e83596.

  50. 50.

    Tinsley JH, Hawker J, Yuan Y. Efficient protein transfection of cultured coronary venular endothelial cells. Am J Physiol-Heart Circ Physiol. 1998;275:H1873–8.

Download references

Acknowledgments and Disclosures

We would like to thank J. Y Exposito and P. Libeau for useful discussions on mRNA aspects of delivery. We are grateful to AMU and CNRS for financial support. Financial support is also gained from ANRS in the framework of HIVERA JTC 2014 (HIV NANOVA), and Euronanomed II (Flunanoair) and from ANR-16-CE20-0002-01 (FishRNAVax) to BV.

Author information

Correspondence to Thomas Trimaille.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material


(DOCX 394 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Lacroix, C., Humanes, A., Coiffier, C. et al. Polylactide-Based Reactive Micelles as a Robust Platform for mRNA Delivery. Pharm Res 37, 30 (2020). https://doi.org/10.1007/s11095-019-2749-6

Download citation


  • cationic fusogenic peptide
  • Micelles
  • polylactide
  • mRNA delivery