I have a strong background in adapting and customising imaging and spatial technologies for research and clinical applications which are crucial for highly heterogenous tissues such as skin. One of my research focuses is to investigate the epidermal innervation of skin in patients suffering from pruritic diseases such as atopic dermatitis and psoriasis. To achieve this, I developed a workflow involving optical clearing, imaging of human skin, and image analysis for nerve filaments. In recent years, I’m interested to work on translating knowledge for disease mechanisms and laboratory techniques to practical solutions for patients, and I am particularly interested to delineate, understand and study therapeutic targeting of the mechanisms driving skin inflammation and itch in clinical patient cohorts.
Abstract submitted for the 5th International Keloid Symposium
Title: Effect of dual siRNAs nanoplex targeting IL-4RA and SPARC in IL-4 activated skin fibroblasts
AUTHORS: Yingrou Tan1,2, Yong Yao Chun3, Caren Lum1,2, Eleanor Shu Xian Chai2,4, Shu Zhen Chong3, Hong Liang Tey1,5 and Timothy Thatt Yang Tan1
AUTHORS’ AFFILIATIONS:
1 National Skin Centre, National Healthcare Group, 1 Mandalay Road, Singapore 308205, Singapore
2 Singapore Immunology Network, 8a Biomedical Grove, Singapore 138648, Singapore.
3 School of Chemistry, Chemical Engineering and Biotechnology, 62 Nanyang Drive, Singapore 637459, Singapore.
4 Duke-NUS Medical School, Singapore 169857
5 Lee Kong Chian School of Medicine, Nanyang Technological University, 11 Mandalay Road, Singapore 308232, Singapore.
Corresponding email: tan_yingrou@immunol.a-star.edu.sg, teyhongliang@ntu.edu.sg,
Background: Currently, there is a need for more effective preventive methods or treatment for hypertrophic scars or keloids. To target important proteins involved in the T helper 2 inflammatory microenvironment and ECM production during scar formation, we designed dual siRNAs targeting interleukin-4 Receptor α (IL-4Rα) and secreted protein acidic and cysteine rich (SPARC) as a novel preventive solution and treatment to reduce pathological scars. In this study, the effect of dual siRNAs (siIL-4Rα and siSPARC) nanoplex in reducing collagen expression for interleukin-4 stimulated human dermal fibroblasts (HDFs) was investigated.
Methods: As unmodified siRNA is easily degraded by endogenous enzymes, we encapsulated the siRNAs using positively charged gelatin-tyramine (Gtn-Tyr) to form a nanoplex to achieve protection and targeted delivery to dermal fibroblast, studied gene knockdown, decrease in collagen production, and compared internalization of the siRNA-nanoplex by dermal fibroblasts with keratinocytes. Simultaneously, we tested the effect of siRNA nanoplex on cell cytotoxicity and proliferation.
Separately, we carried out a case series in three individuals with hypertrophic scars and keloid who volunteered. The dual siRNA-nanoplex delivered via microneedles for treatment and their scars were assessed at different timepoints.
Results: The results demonstrated that siRNA-nanoplex preferentially enhances fibroblast uptake in comparison to naked siRNA, and when compared with keratinocytes. Majority of the siRNA-nanoplex is also not present within lysosomal associated membrane protein-1 (LAMP-1)-positive late endosomes for degradation even after 24h of treatment. Simultaneously, preliminary data showed that the dual siRNAs nanoplex of siIL-4Rα and siSPARC could inhibit collagen production of HDFs in the presence of IL-4 at the transcriptional level, and dual siRNAs nanoplex treatment does not cause any cytotoxic effect on cells or inhibit cell proliferation.
Volunteers that tested the microneedle delivered dual siRNA-nanoplex had reduction of scar volume together with reduction of pain or itch, with no adverse effects reported.
Conclusion: Overall, targeting both IL-4Rα and SPARC using the siRNA nanoplex demonstrates a promising solution for preventing and treating hypertrophic scars or keloids.
Presenting Author:
Name: Tan Yingrou
Academic Degree: Doctor of Philosophy
Institution: National Skin Centre,
Address: 1 Mandalay Road, Singapore 308205
Email address: tan_yingrou@immunol.a-star.edu.sg,
Category:
Original Research
