Topical application of proteins as a new therapy option for the treatment of severe, genetic skin diseases
Dermatology
Final Report Abstract
Topical protein delivery to the skin is not usually possible owing to the exceptional barrier proteins of the skin’s outer most layer, the stratum corneum. However, the inherent barrier deficiencies seen in many skin diseases may be manipulated to permit the passage of potentially therapeutic macromolecules. This project builds upon previous work in which thermoresponsive nanogels (tNG) were used as a novel intervention to deliver therapeutic proteins to the viable epidermis of diseased skin. In a first step, tNGs composed of dendriticpoly(glycerol) interlinked by poly(N-Isopropylacrylamide) (dPG-pNIPAM) were used to encapsulate transglutaminase 1 (TG1) and deliver this to full thickness reconstructed human skin – skin equivalents – formed from the keratinocytes of patients possessing a monogenic skin disease caused by loss of TG1 function. TG1 plays an essential role in the formation of the cornified envelope, a structure key to the skin’s barrier properties. Treatment with TG1 loaded tNGs was shown to deliver functionally active TG1 to the stratum corneum and late viable epidermis, and to improve the characteristic skin barrier deficiencies seen in these skin equivalents in a dose dependent manner. Toxicity studies demonstrated the excellent biocompatibility of TG1 loaded tNGs in isolated primary human skin cells and fully formed skin equivalents. Work with monocytes derived immune cells also showed the excellent immune-compatibility of these and a second tNG chemistry based on dendritic polyglycerol interlinked by thermoresponsive polyglycerol (dPG-tPG). To expand the potential therapeutic relevance of topical protein delivery, the TNFα sequestering protein Etanercept (ETR) – currently approved for the treatment of psoriasis by subcutaneous administration – was encapsulated within the dPG-pNIPAM and dPG-tPG tNGs. This was shown to enhance the long term activity of ETR. Temperature triggered protein release was seen for both tNGs, and topical application of either resulted in ETR delivery to the viable epidermis of inflamed skin equivalents that correlated with antiinflammatory effects. The tNGs were also shown to deliver protein into tape stripped human skin biopsies – a validated model of barrier deficient skin – in a manner that appeared dependent on their temperature induced protein release. The results of the project demonstrate the potential therapeutic utilities of protein replacement strategies in monogenic skin diseases and delivery of cytokine sequestering proteins in inflammatory skin conditions. The tractability of such interventions would be greatly enhanced by a topical mode of application, something the tNGs were able to accomplish in a highly biocompatible manner. The use of tNGs as a platform technology for the topical application of macromolecules holds real clinical promise for multiple skin conditions and work to optimise their efficiency of epidermal protein delivery, and thereby their therapeutic efficacy, should be the next focus of investigation.
Publications
- (2019) Transglutaminase 1 Replacement Therapy Successfully Mitigates the Autosomal Recessive Congenital Ichthyosis Phenotype in Full-Thickness Skin Disease Equivalents. The Journal of investigative dermatology 139 (5) 1191–1195
Plank, Roswitha; Yealland, Guy; Miceli, Enrico; Lima Cunha, Dulce; Graff, Patrick; Thomforde, Sari; Gruber, Robert; Moosbrugger-Martinz, Verena; Eckl, Katja; Calderón, Marcelo; Hennies, Hans Christian; Hedtrich, Sarah
(See online at https://doi.org/10.1016/j.jid.2018.11.002) - Feasibility study for intraepidermal delivery of proteins using a solid microneedle array. International Journal of Pharmaceutics.2015; 486(1-2): 52-58
Witting M, Obst K, Pietzsch M, Friess W, Hedtrich S
(See online at https://doi.org/10.1016/j.ijpharm.2015.03.046) - Interactions of Hyaluronic Acid with the Skin and Implications for the Dermal Delivery of Biomacromolecules. Molecular Pharmaceutics. 2015; 12(5): 1391-1401
Witting M, Boreham A, Brodwolf R, Vávrová K, Alexiev U, Friess W, Hedtrich S
(See online at https://doi.org/10.1021/mp500676e) - Recent advances in topical delivery of proteins and peptides mediated by soft matter nanocarriers. Biotechnology Advances. 2015; 33(6): 1355-1369
Witting M, Obst K, Friess W, Hedtrich S
(See online at https://doi.org/10.1016/j.biotechadv.2015.01.010) - Thermosensitive dendritic polyglycerol-based nanogels for cutaneous delivery of biomacromolecules. Nanomedicine: Nanotechnology, Biology and Medicine. 2015; 11: 1179-87
Witting M, Molina M, Obst K, Plank R, Eckl KM, Hennies HC et al.
(See online at https://doi.org/10.1016/j.nano.2015.02.017) - Protein Corona Formation on Colloidal Polymeric Nanoparticles and Polymeric Nanogels: Impact on Cellular Uptake, Toxicity, Immunogenicity, and Drug Release Properties. Biomacromolecules. 2017; 18(6): 1762-1771
Obst K, Yealland G, Balzus B, Miceli E, Dimde M, Weise C, Eravci M, Bodmeier R, Haag R, Calderón M, Charbaji N, Hedtrich S
(See online at https://doi.org/10.1021/acs.biomac.7b00158) - Breaking the Barrier - Potent Anti-Inflammatory Activity following Efficient Topical Delivery of Etanercept using Thermoresponsive Nanogels. Theranostics. 2018; 8(2):450-463
Giulbudagian M, Yealland G, Hönzke S, Edlich A, Geisendörfer B, Kleuser B, Hedtrich S, Calderón M
(See online at https://doi.org/10.7150/thno.21668)
