Exploring combinatorial microRNA therapy for chronic wound healing
File(s)
Author(s)
Pop, Mara Alexandra
Type
Thesis
Abstract
Wound healing is a highly orchestrated process involving the coordinated action of many cell types, growth factors, cytokines and small regulatory RNAs, such as microRNAs (miRNAs). These small RNA molecules become expressed in a spatiotemporal pattern to fine-tune signalling pathways coordinating tissue repair. Recently, miRNA dysregulation has been linked to impaired healing, including the formation and progression of diabetic foot ulcers (DFUs), a type of chronic wound associated with diabetes. While most of the interest has gone into the role of individual miRNAs in DFUs, research into the combinatorial role of dysregulated miRNAs remains limited. Complicating factors, such as dosage-dependent effects of miRNAs and current stochastic delivery methods, can make it challenging to study combinatorial miRNA effects.
This project addressed some of these understudied limiting elements to explore how combinatorial miRNA dysregulation can contribute to impaired healing in DFUs. First, a set of differentially expressed miRNAs between DFU-derived fibroblasts and healthy fibroblasts was identified and, using bioinformatics, the biological impact of their dysregulation was explored in delaying wound healing. Next, three miRNAs were chosen from this dataset and their individual and combinatorial effects was investigated in primary skin cells isolated from amputated human limbs. Lastly, a novel in vitro engineering-based approach for miRNA delivery was used for studying incremental dosage effects of specific differentially expressed miRNAs. Altogether, these findings opened opportunities for investigating dose-dependent effects of miRNA combinations for the treatment of DFUs, yet not addressed by current research.
This project addressed some of these understudied limiting elements to explore how combinatorial miRNA dysregulation can contribute to impaired healing in DFUs. First, a set of differentially expressed miRNAs between DFU-derived fibroblasts and healthy fibroblasts was identified and, using bioinformatics, the biological impact of their dysregulation was explored in delaying wound healing. Next, three miRNAs were chosen from this dataset and their individual and combinatorial effects was investigated in primary skin cells isolated from amputated human limbs. Lastly, a novel in vitro engineering-based approach for miRNA delivery was used for studying incremental dosage effects of specific differentially expressed miRNAs. Altogether, these findings opened opportunities for investigating dose-dependent effects of miRNA combinations for the treatment of DFUs, yet not addressed by current research.
Version
Open Access
Date Issued
2020-01
Date Awarded
2020-06
Copyright Statement
Creative Commons Attribution NonCommercial NoDerivatives Licence
Advisor
Almquist, Benjamin
Publisher Department
Bioengineering
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)