Targeted delivery of LM22A-4 by cubosomes protects retinal ganglion cells in an experimental glaucoma model

Yue Ding; Seong H Chow; Jinying Chen; Anton P Le Brun; Chun-Ming Wu; Anthony P Duff; Yajun Wang; Jiangning Song; Jiang-Hui Wang; Vickie H Y Wong; Da Zhao; Tomoharu Nishimura; Tzong-Hsien Lee; Charlotte E Conn; Hsien-Yi Hsu; Bang V Bui; Guei-Sheung Liu; Hsin-Hui Shen

doi:10.1016/j.actbio.2021.03.043

Targeted delivery of LM22A-4 by cubosomes protects retinal ganglion cells in an experimental glaucoma model

Acta Biomater. 2021 May:126:433-444. doi: 10.1016/j.actbio.2021.03.043. Epub 2021 Mar 24.

Authors

Yue Ding¹, Seong H Chow², Jinying Chen³, Anton P Le Brun⁴, Chun-Ming Wu⁵, Anthony P Duff⁴, Yajun Wang⁶, Jiangning Song², Jiang-Hui Wang⁷, Vickie H Y Wong⁸, Da Zhao⁸, Tomoharu Nishimura⁸, Tzong-Hsien Lee², Charlotte E Conn⁹, Hsien-Yi Hsu¹⁰, Bang V Bui⁷, Guei-Sheung Liu¹¹, Hsin-Hui Shen¹²

Affiliations

¹ Department of Materials Science and Engineering, Monash University, Clayton, VIC 3800, Australia; Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, VIC 3800, Australia.
² Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, VIC 3800, Australia.
³ Menzies Institute for Medical Research, University of Tasmania, Hobart, TAS 7000, Australia; Department of Ophthalmology, the First Affiliated Hospital of Jinan University, Guangzhou, Guangdong 510632, China.
⁴ Australian Nuclear Science and Technology Organization (ANSTO), Lucas Heights, NSW 2234, Australia.
⁵ Australian Nuclear Science and Technology Organization (ANSTO), Lucas Heights, NSW 2234, Australia; National Synchrotron Radiation Research Center, Hsinchu 30076, Taiwan.
⁶ College of Chemistry and Materials Engineering, Wenzhou University, Wenzhou, Zhejiang 325035, China.
⁷ Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital, East Melbourne, VIC 3002, Australia.
⁸ Department of Optometry and Vision Sciences, University of Melbourne, Parkville, VIC 3052, Australia.
⁹ School of Science, College of Science, Engineering and Health, RMIT University, Melbourne, VIC 3000, Australia.
¹⁰ School of Energy and Environment & Department of Materials Science and Engineering, City University of Hong Kong, Kowloon Tong, Hong Kong, China; Shenzhen Research Institute of City University of Hong Kong, Shenzhen, Guangdong 518057, China.
¹¹ Menzies Institute for Medical Research, University of Tasmania, Hobart, TAS 7000, Australia; Ophthalmology, Department of Surgery, University of Melbourne, East Melbourne, VIC 3002, Australia; Aier Eye Institute, Changsha, Hunan 410015, China. Electronic address: rickliu0817@gmail.com.
¹² Department of Materials Science and Engineering, Monash University, Clayton, VIC 3800, Australia; Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, VIC 3800, Australia. Electronic address: Hsin-Hui.Shen@monash.edu.

PMID: 33774200
DOI: 10.1016/j.actbio.2021.03.043

Abstract

Glaucoma, a major cause of irreversible blindness worldwide, is associated with elevated intraocular pressure (IOP) and progressive loss of retinal ganglion cells (RGCs) that undergo apoptosis. A mechanism for RGCs injury involves impairment of neurotrophic support and exogenous supply of neurotrophic factors has been shown to be beneficial. However, neurotrophic factors can have widespread effects on neuronal tissues, thus targeting neurotrophic support to injured neurons may be a better neuroprotective strategy. In this study, we have encapsulated LM22A-4, a small neurotrophic factor mimetic, into Annexin V-conjugated cubosomes (L4-ACs) for targeted delivery to injured RGCs in a model of acute IOP elevation, which is induced by acute IOP elevation. We have tested cubosomes formulations that encapsulate from 9% to 33% LM22A-4. Our data indicated that cubosomes encapsulating 9% and 17% LM22A-4 exhibited a mixture of Pn3m/Im3m cubic phase, whereas 23% and 33% showed a pure Im3m cubic phase. We found that 17% L4-ACs with Pn3m/Im3m symmetries showed better in-situ and in-vitro lipid membrane interactions than the 23% and 33% L4-ACs with Im3m symmetry. In vivo experiments showed that 17% L4-ACs targeted the posterior retina and the optic nerve head, which prevented RGCs loss and improved functional outcomes in a mouse model of acute IOP elevation. These results provide evidence that Annexin V-conjugated cubosomes-based LM22A-4 delivery may be a useful targeted approach to prevent the progression of RGCs loss in glaucoma. STATEMENT OF SIGNIFICANCE: Recent studies suggest that the therapy of effectively delivering neurotrophic factors to the injured retinal ganglion cells (RGCs) could promote the survival of RGCs in glaucoma. Our present work has for the first time used cubosomes as an active targeted delivery system and have successfully delivered a neuroprotective drug to the damaged RGCs in vivo. Our new cubosomal formulation can protect apoptotic cell death in vitro and in vivo, showing that cubosomes are a promising drug carrier system for ocular drug delivery and glaucoma treatment. We have further found that by controlling cubosomes in Pn3m phase we can facilitate delivery of neuroprotective drug through apoptotic membranes. This data, we believe, has important implications for future design and formulation of cubosomes for therapeutic applications.

Keywords: Apoptosis; Cubosomes; Intraocular pressure and experimental glaucoma model; LM22A-4; Neutron reflectometry; QCM-D; Retinal ganglion cell; SAXS.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Benzamides
Disease Models, Animal
Glaucoma* / drug therapy
Intraocular Pressure
Mice
Optic Disk*
Retinal Ganglion Cells

Substances

Benzamides
N,N',N'-tris(2-hydroxyethyl)-1,3,5-benzenetricarboxamide