Research Article

Department of Chemical & Biomolecular Engineering, Center for Nanotherapeutic Strategies in the Central Nervous System, Clemson University, Clemson, SC 29631, USA

Search for more papers by this author

Alaura Cakley

Department of Chemical & Biomolecular Engineering, Center for Nanotherapeutic Strategies in the Central Nervous System, Clemson University, Clemson, SC 29631, USA

Search for more papers by this author

Jessica Larsen

*Author for correspondence:

E-mail Address: larsenj@clemson.edu

https://orcid.org/0000-0003-2756-9523

Department of Chemical & Biomolecular Engineering, Center for Nanotherapeutic Strategies in the Central Nervous System, Clemson University, Clemson, SC 29631, USA

Department of Bioengineering, Center for Nanotherapeutic Strategies in the Central Nervous System, Clemson University, Clemson, SC 29631, USA

Search for more papers by this author

Published Online:25 Jan 2024https://doi.org/10.2217/nnm-2023-0300

View article

Abstract

Aims: Stimuli-responsive polymersomes are promising tools for protein-based therapies, but require deeper understanding and optimization of their pathology-responsive behavior. Materials & methods: Hyaluronic acid (HA)–poly(b-lactic acid) (PLA) polymersomes self-assembled from block copolymers of varying molecular weights of HA were compared for their physical properties, degradation and intracellular behavior. Results: Major results showed increasing enzyme-responsivity associated with decreasing molecular weight. The major formulation differences were as follows: the HA(5 kDa)–PLA formulation exhibited the most pronounced release of encapsulated proteins, while the HA(7 kDa)–PLA formulation showed the most different release behavior from neutral. Conclusion: We have discovered design rules for HA–PLA polymersomes for protein delivery, with lower molecular weight leading to higher encapsulation efficiency, greater release and greater intracellular uptake.

Keywords:

Papers of special note have been highlighted as: • of interest; •• of considerable interest

References

1. Ebrahimi SB, Samanta D. Engineering protein-based therapeutics through structural and chemical design. Nat. Commun. 14(1), 2411 (2023).
Medline CASGoogle Scholar
2. Leader B, Quentin B, Golan D. Protein therapeutics: a summary and pharmacological classification. Nat. Rev. Drug. Discov. (7), 21–39 (2008).
MedlineGoogle Scholar
3. Gorzelany J, Souza M. Protein replacement therapies for rare diseases: a breeze for regulatory approval? Sci. Transl. Med. 5(178), 10.1126/scitranslmed.3005007 (2013).
MedlineGoogle Scholar
4. Patel A, Cholkar K, Mitra AK. Recent developments in protein and peptide parenteral delivery approaches. Ther. Deliv. 5(3), 337–365 (2014).
Medline CASGoogle Scholar
5. Kelly JM, Gross AL, Martin DR, Byrne ME. Polyethylene glycol-b-poly(lactic acid) polymersomes as vehicles for enzyme replacement therapy. Nanomedicine 12(23), 2591–2606 (2017).
CASGoogle Scholar
6. Iqbal S, Blenner M, Alexander-Bryant A, Larsen J. Polymersomes for therapeutic delivery of protein and nucleic acid macromolecules: from design to therapeutic applications. Biomacromolecules 21(4), 1327–1350 (2020).
Medline CASGoogle Scholar
7. Discher DE, Ahmed F. Polymersomes. Annu. Rev. Biomed. Eng. 8, 323–341 (2006). • A founding source of information on polymersomes; the paper was important to the material choices and evaluation procedures used here.
Medline CASGoogle Scholar
8. Zhang A, Jung K, Li A, Liu J, Boyer C. Recent advances in stimuli-responsive polymer systems for remotely controlled drug release. Prog. Polym. Sci. 99, 101164 (2019).
CASGoogle Scholar
9. Kost J, Langer R. Responsive polymeric delivery systems. Adv. Drug Deliv. Rev. 64, 327–341 (2012).
Google Scholar
10. Meng F, Zhong Z, Feijen J. Stimuli-responsive polymersomes for programmed drug delivery. Biomacromolecules 10(2), 197–209 (2009).
Medline CASGoogle Scholar
11. Wells CM, Harris M, Choi L, Murali VP, Guerra FD, Jennings JA. Stimuli-responsive drug release from smart polymers. J. Funct. Biomater. 10(3), 34 (2019).
Medline CASGoogle Scholar
12. Kaul A, Short WD, Wang X, Keswani SG. Hyaluronidases in human diseases. Int. J. Mol. Sci. 22(6), 3204 (2021).
Medline CASGoogle Scholar
13. Hu Q, Katti PS, Gu Z. Enzyme-responsive nanomaterials for controlled drug delivery. Nanoscale 6(21), 12273–12286 (2014).
Medline CASGoogle Scholar
14. Srinivasarao M, Low PS. Ligand-targeted drug delivery. Chem. Rev. 117(19), 12133–12164 (2017).
Medline CASGoogle Scholar
15. Yu MK, Park J, Jon S. Targeting strategies for multifunctional nanoparticles in cancer imaging and therapy. Theranostics 2(1), 3–44 (2012).
Medline CASGoogle Scholar
16. van der Meel R, Vehmeijer LJC, Kok RJ, Storm G, van Gaal EVB. Ligand-targeted particulate nanomedicines undergoing clinical evaluation: current status. Adv. Drug Deliv. Rev. 65(10), 1284–1298 (2013).
MedlineGoogle Scholar
17. Wang M, Thanou M. Targeting nanoparticles to cancer. Pharmacol. Res. 62(2), 90–99 (2010).
Medline CASGoogle Scholar
18. Storr T, Thompson KH, Orvig C. Design of targeting ligands in medicinal inorganic chemistry. Chem. Soc. Rev. 35(6), 534–544 (2006).
Medline CASGoogle Scholar
19. Tan JX, Wang XY, Su XL et al. Upregulation of HYAL1 expression in breast cancer promoted tumor cell proliferation, migration, invasion and angiogenesis. PLOS ONE 6(7), 10.1371/journal.pone.0022836 (2011).
Google Scholar
20. Yoffou PH, Edjekouane L, Meunier L et al. Subtype specific elevated expression of hyaluronidase-1 (HYAL-1) in epithelial ovarian cancer. PLOS ONE 6(6), 10.1371/journal.pone.0020705 (2011).
MedlineGoogle Scholar
21. Raj P, Lee SY, Lee TY. Carbon dot/naphthalimide based ratiometric fluorescence biosensor for hyaluronidase detection. Materials 14(5), 10.3390/ma14051313 (2021).
Google Scholar
22. Paruchuri BC, Smith S, Larsen J. Enzyme-responsive polymersomes ameliorate autophagic failure in a cellular model of GM1 gangliosidosis. Front. Chem. Eng. 4, 91 (2022). •• Serves as the basis for the system which our current research paper worked to optimize; much of our study protocol follows what was set forth in this paper.
Google Scholar
23. Meszaros M, Kis A, Kunos L et al. The role of hyaluronic acid and hyaluronidase-1 in obstructive sleep apnoea. Sci. Rep. 10(1), 10.1038/s41598-020-74769-4 (2020).
Google Scholar
24. Albeiroti S, Ayasoufi K, Hill DR, Shen B, De La Motte CA. Platelet hyaluronidase-2: an enzyme that translocates to the surface upon activation to function in extracellular matrix degradation. Blood 125(9), 1460–1469 (2015).
Medline CASGoogle Scholar
25. Zhang L, Bharadwaj AG, Casper A, Barkley J, Barycki JJ, Simpson MA. Hyaluronidase activity of human Hyal1 requires active site acidic and tyrosine residues. J. Biol. Chem. 284(14), 9433–9442 (2009).
Medline CASGoogle Scholar
26. Boedtkjer E, Pedersen SF. The acidic tumor microenvironment as a driver of cancer. Annu. Rev. Physiol. 82, 103–126 (2020).
Medline CASGoogle Scholar
27. Reshetnyak YK, Yao L, Zheng S, Kuznetsov S, Engelman DM, Andreev OA. Measuring tumor aggressiveness and targeting metastatic lesions with fluorescent pHLIP. Mol. Imaging Biol. 13(6), 1146–1156 (2011).
MedlineGoogle Scholar
28. Feng L, Dong Z, Tao D, Zhang Y, Liu Z. The acidic tumor microenvironment: a target for smart cancer nano-theranostics. National Sci. Rev. 5(2), 269–286 (2018).
CASGoogle Scholar
29. Platt FM. Emptying the stores: lysosomal diseases and therapeutic strategies. Nat. Rev. Drug Discov. 17(2), 133–150 (2018).
Medline CASGoogle Scholar
30. Paruchuri BC, Smith S, Larsen J. Enzyme-responsive polymersomes ameliorate autophagic failure in a cellular model of GM1 gangliosidosis. Front. Chem. Eng. 4, 91 (2022).
Google Scholar
31. Tyler B, Gullotti D, Mangraviti A, Utsuki T, Brem H. Polylactic acid (PLA) controlled delivery carriers for biomedical applications. Adv. Drug Deliv. Rev. 107, 163–175 (2016).
Medline CASGoogle Scholar
32. Casalini T, Rossi F, Castrovinci A, Perale G. A perspective on polylactic acid-based polymers use for nanoparticles synthesis and applications. Front. Bioeng. Biotechnol. 7, 259 (2019).
MedlineGoogle Scholar
33. Lee BM, Park SJ, Noh I, Kim CH. The effects of the molecular weights of hyaluronic acid on the immune responses. Biomater. Res. 25(1), https://doi.org/10.1186/s40824-021-00228-4 (2021).
Google Scholar
34. Dovedytis M, Liu ZJ, Bartlett S. Hyaluronic acid and its biomedical applications: a review. Eng. Regener. 1, 102–113 (2020).
Google Scholar
35. Snetkov P, Zakharova K, Morozkina S, Olekhnovich R, Uspenskaya M. Hyaluronic acid: the influence of molecular weight on structural, physical, physico-chemical, and degradable properties of biopolymer. Polymers (Basel) 12(8), 10.3390/polym12081800 (2020). • Paramount in explaining and understanding our observed results as we compared the impact of hyaluronic acid’s molecular weight in our own system specifically.
MedlineGoogle Scholar
36. Anselmo AC, Mitragotri S. Nanoparticles in the clinic: an update post COVID-19 vaccines. Bioeng. Transl. Med. 6(3), 1–20 (2021).
Google Scholar
37. Han E, Kim D, Cho Y, Lee S, Kim J, Kim H. Development of polymersomes co-delivering doxorubicin and melittin to overcome multidrug resistance. Molecules 28(3), 1087 (2023).
Medline CASGoogle Scholar
38. Tücking KS, Grützner V, Unger RE, Schönherr H. Dual enzyme-responsive capsules of hyaluronic acid-block-poly(lactic acid) for sensing bacterial enzymes. Macromol. Rapid Commun. 36(13), 1248–1254 (2015).
Medline CASGoogle Scholar
39. Paap MK, Silkiss RZ. The interaction between hyaluronidase and hyaluronic acid gel fillers – a review of the literature and comparative analysis. Plast. Aesthet. Res. 7, 36 (2020).
Google Scholar
40. Stern R, Kogan G, Jedrzejas MJ, Šoltés L. The many ways to cleave hyaluronan. Biotechnol. Adv. 25(6), 537–557 (2007).
Medline CASGoogle Scholar
41. Gushulak L, Hemming R, Martin D, Seyrantepe V, Pshezhetsky A, Triggs-Raine B. Hyaluronidase 1 and β-hexosaminidase have redundant functions in hyaluronan and chondroitin sulfate degradation. J. Biol. Chem. 287(20), 16689 (2012).
Medline CASGoogle Scholar
42. Tømmeraas K, Melander C. Kinetics of hyaluronan hydrolysis in acidic solution at various pH values. Biomacromolecules 9(6), 1535–1540 (2008).
MedlineGoogle Scholar
43. Casalini T, Rossi F, Castrovinci A, Perale G. A perspective on polylactic acid-based polymers use for nanoparticles synthesis and applications. Front. Bioeng. Biotechnol. 7, 259 (2019).
MedlineGoogle Scholar
44. Sha L, Chen Z, Chen Z, Zhang A, Yang Z. Polylactic acid based nanocomposites: promising safe and biodegradable materials in biomedical field. Int. J. Polym. Sci. 2016, 6869154 (2016).
Google Scholar
45. Deng C, Xu X, Tashi D, Wu Y, Su B, Zhang Q. Co-administration of biocompatible self-assembled polylactic acid–hyaluronic acid block copolymer nanoparticles with tumor-penetrating peptide-iRGD for metastatic breast cancer therapy. J. Mater. Chem. B 6(19), 3163–3180 (2018).
Medline CASGoogle Scholar
46. Kelly JM, Pearce EE, Martin DR, Byrne ME. Lyoprotectants modify and stabilize self-assembly of polymersomes. Polymer 87, 316–322 (2016).
CASGoogle Scholar
47. Casal JA, Cano E, Tutor JC. β-Hexosaminidase isoenzyme profiles in serum, plasma, platelets and mononuclear, polymorphonuclear and unfractionated total leukocytes. Clin. Biochem. 38(10), 938–942 (2005).
Medline CASGoogle Scholar
48. Kelly JM, Bradbury A, Martin DR, Byrne ME. Emerging therapies for neuropathic lysosomal storage disorders. Prog. Neurobiol. 152, 166–180 (2017).
Medline CASGoogle Scholar
49. Chang HY, Sheng YJ, Tsao HK. Structural and mechanical characteristics of polymersomes. Soft Matter 10(34), 6373–6381 (2014).
Medline CASGoogle Scholar
50. Holowka EP, Pochan DJ, Deming TJ. Charged polypeptide vesicles with controllable diameter. J. Am. Chem. Soc. 127(35), 12423–12428 (2005).
Medline CASGoogle Scholar
51. Krook NM, Tabedzki C, Elbert KC et al. Experiments and simulations probing local domain bulge and string assembly of aligned nanoplates in a lamellar diblock copolymer. Macromolecules 52(22), 8989–8999 (2019).
CASGoogle Scholar
52. Gouveia MG, Wesseler JP, Ramaekers J, Weder C, Scholten PBV, Bruns N. Polymersome-based protein drug delivery – quo vadis? Chem. Soc. Rev. 52(2), 728–778 (2022).
Google Scholar
53. Joye I, McClements D. Biopolymer-based delivery systems: challenges and opportunities. Curr. Top Med. Chem. 16(9), 1026–1039 (2016).
Medline CASGoogle Scholar
54. Furtado D, Björnmalm M, Ayton S, Bush AI, Kempe K, Caruso F. Overcoming the blood–brain barrier: the role of nanomaterials in treating neurological diseases. Adv. Mater. 30(46), e1801362 (2018).
MedlineGoogle Scholar
55. Buhren BA, Schrumpf H, Bölke E, Kammers K, Gerber PA. Standardized in vitro analysis of the degradability of hyaluronic acid fillers by hyaluronidase. Eur. J. Med. Res. 23(1), 1–6 (2018).
MedlineGoogle Scholar
56. Kim SW, Oh KT, Youn YS, Lee ES. Hyaluronated nanoparticles with pH- and enzyme-responsive drug release properties. Colloids Surf. B Biointerfaces 116, 359–364 (2014).
Medline CASGoogle Scholar
57. Biomol GmbH. Guide to enzyme unit definitions and assay design (2019). www.biomol.com/resources/biomol-blog/guide-to-enzyme-unit-definitions-and-assay-design
Google Scholar
58. Sigma Aldrich. Hyaluronidase type II, lyophilized powder, main=300units/mg solid 37326-33-3. www.sigmaaldrich.com/US/en/product/sigma/h2126
Google Scholar
59. Lee JS, Feijen J. Polymersomes for drug delivery: design, formation and characterization. J. Control. Rel. 161(2), 473–483 (2012).
Medline CASGoogle Scholar
60. ISO. 10993-5: 2009 Biological evaluation of medical devices-part 5: tests for in vitro cytotoxicity (2009). www.iso.org/standard/36406.html
Google Scholar
61. Luckanagul JA, Bhuket PRN, Muangnoi C, Rojsitthisak P, Wang Q, Rojsitthisak P. Self-assembled thermoresponsive nanogel from grafted hyaluronic acid as a biocompatible delivery platform for curcumin with enhanced drug loading and biological activities. Polymers (Basel) 13(2), 1–14 (2021).
Google Scholar
62. Deng C, Xu X, Tashi D, Wu Y, Su B, Zhang Q. Co-administration of biocompatible self-assembled polylactic acid-hyaluronic acid block copolymer nanoparticles with tumor-penetrating peptide-iRGD for metastatic breast cancer therapy. J. Mater. Chem. B 6(19), 3163–3180 (2018).
Medline CASGoogle Scholar
63. Alipoor R, Ayan M, Hamblin MR, Ranjbar R, Rashki S. Hyaluronic acid-based nanomaterials as a new approach to the treatment and prevention of bacterial infections. Front. Bioeng. Biotechnol. 10, 913912 (2022).
MedlineGoogle Scholar
64. Gheran CV, Rigaux G, Callewaert M et al. Biocompatibility of Gd-loaded chitosan–hyaluronic acid nanogels as contrast agents for magnetic resonance cancer imaging. Nanomaterials 8(4), 10.3390/nano8040201 (2018).
MedlineGoogle Scholar
65. Nance EA, Woodworth GF, Sailor KA et al. A dense poly(ethylene glycol) coating improves penetration of large polymeric nanoparticles within brain tissue. Sci. Transl. Med. 4(149), 10.1126/scitranslmed.3003594 (2012).
MedlineGoogle Scholar

Vol. 19, No. 3

Supplemental Materials

Metrics

Downloaded 33 times

History

Received 16 October 2023

Accepted 20 November 2023

Published online 25 January 2024

Published in print February 2024

Information

Keywords

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.futuremedicine.com/doi/suppl/10.2217/nnm-2023-0300

Author contributions

D Foster and J Larsen were responsible for the conception and design of experiments, analysis and interpretation of the data and revising this work critically for intellectual content. D Foster was responsible for conducting experiments and drafting of the paper; A Cakley assisted in conducting experiments. All authors approved the final version to be published; and agree to be accountable for all aspects of the work.

Acknowledgments

The authors thank D Martin and A Gross at Auburn University for the use of the GM1SV3 cell lines and E Davis for the use of ATR-FTIR spectrophotometer.

Financial disclosure

This work was supported in part by the National Science Foundation CAREER program under NSF Award no. 2047697. Support was also received from Clemson University’s Creative Inquiry program. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

Competing interests disclosure

The authors have no competing interests or relevant affiliations with any organization or entity with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

Writing disclosure

No writing assistance was utilized in the production of this manuscript.

PDF download

Optimizing enzyme-responsive polymersomes for protein-based therapies

Abstract

References