Periodic Reporting for period 1 - miRaColl (miRNA-activated Scaffold Technologies for Cartilage Regeneration)
Période du rapport: 2015-09-01 au 2017-02-28
Résumé du contexte et des objectifs généraux du projet
Collectively, this research programme has been successful and centred on development of microRNA-activated collagen-based materials developed in Prof. O’Brien’s laboratory to address the particularly difficult problem of enhanced cartilage repair. It has utilised new insight and technologies including research in biomaterials science, gene therapy, stem cell biology, high-throughput gene screening and bioengineering through the inclusion of miRNAs to augment the healing profile of the scaffolds and ultimately develop a superior, bioactive scaffold for cartilage regeneration. Specific outputs from the different themes are as follows:
WP1: In vitro development and characterisation of miRNA-activated collagen (miRaColl) scaffolds
The primary outcome of this work package was the successful development and characterisation of functional miRNA-activated collagen (miRaColl) scaffolds. miRNA dose and delivery potential of various in-house developed delivery vectors including nanohydroxyapatite, polyethyleneimine and oligomeric chitosan were analysed in comparison to the commercial gold standard Lipofectamine RNAiMAX, and characterised using a combination of techniques in line with those optimised previously in Prof. O’Brien’s laboratory as part of a study on miRNA-activated scaffolds for bone repair. A particularly low dose of Lipofectamine RNAiMAX was optimised compared to doses previously reported in the literature, which resulted in the optimised Lipo-miRNA formulation being brought forward for further testing on our in-house developed collagen-hyaluronic acid scaffolds optimised specifically for cartilage-repair. miRNA levels were effectively manipulated across two microRNAs when assessed in MSC-seeded scaffolds. These specific targets were chosen based on the most recent evidence in the literature.
WP2: Testing and analysis of therapeutic miRaColl scaffolds in vitro using MSCs
Work undertaken in this theme demonstrated development of a novel therapeutic miRNA-activated scaffold but unsurprisingly, significant delays were experienced owing to technical issues encountered when optimising the system, due to this being a very nascent area of research. Firstly, this work focussed on testing functionality and therapeutic capacity of the miRNA-activated scaffolds containing miRNA-mimics, chosen based on the most recent evidence in the literature as indicated in WP1, on four human MSC donors. In parallel, we sought to identify additional chondrogenic miRNA targets of relevance within the three-dimensional environment of our in-house developed collagen-hyaluronic acid scaffolds due to the issues that arose while optimising the system. In order to do so, a high-throughput screening across 768 miRNAs was carried out identifying a remarkably novel miRNA. A comprehensive validation of the chondrogenic role of this novel miRNA is currently underway. Overall, the work undertaken in this theme demonstrated that miRaColl scaffolds enhanced expression of a range of cartilage-specific markers in comparison with the untreated controls. Cartilage matrix deposition analysis of miRaColl scaffolds revealed significant increases in sulphated glycosaminoglycan deposition, the hallmark constituent indicative of MSC chondrogenesis, when compared to relevant controls, pointing to the excellent potential of miRaColl to become a ground-breaking therapeutic with immense commercial interest.
WP3: Assessment of the miRaColl scaffold to regenerate full-thickness chondral defects in vivo
The work performed on this theme encountered delays due to technical difficulties incurred in WP2. However, as the optimal miRaColl system has now been identified, this promising miRaColl system has moved forward and in vivo work is currently ongoing; we are hopeful that a very positive result will be obtained in the coming months.
WP4: Commercial, regulatory and exploitation feasibility analysis of the miRaColl scaffold
The delays encountered in WP2 focusing on optimising the miRaColl system slowed down this commercialisation work package but since the successful miRaColl system has been identified and characterised, this work has proceeded to IDF submission and initial patentability analysis has confirmed that it is worthy of patent filing. As a whole, the thorough analysis conducted in WPs 1-3 has led to the development of a much more marketable technology that appears very promising. Relevant exploitation feasibility analysis is currently ongoing and some positive engagement with commercial partners has been conducted to-date which holds potential for the future.
Quantitative Output
To summarise some of the quantitative output from the project: 1 student completed a research Masters on the project (MSc in Bioengineering) and 1 student completed a summer research project. A number of publications are under preparation that are related to the project and will be published in leading peer-reviewed journals: 1 invited Progress Report focusing on “microRNA therapies in regenerative medicine” in Advanced Healthcare Materials; 1 invited Chapter on miRNA Modulation in Tissue Engineering and Regeneration: Cell Engineering and Regeneration.
Additionally, interactions with industry delegates at a number of events have resulted in positive engagement for progression of the commercialisation aspect of the project, including the 10th International MicroRNAs/Non-Coding RNAs & Genome Editing Europe 2015 Meeting held in Cambridge, UK, organised by GeneExpression Systems, Inc; the Controlled Release Society 2015 Annual Meeting held in Edinburgh, UK; the RNA Therapeutics - Advances and Opportunities 1st Innovation Workshop 2015 held in RCSI Dublin; the Gordon Research Conference on Musculoskeletal Biology and Bioengineering 2016 held in Andover, New Hampshire, US and the Exiqon microRNA & non-coding RNA Seminar 2016 held in Conway Institute UCD Dublin.
The research has been broadly covered by the media without compromising the commercialization element of the project and we have provided extensive opportunities for the next generation of researchers in the form of undergraduates, primary school and secondary school students and their teachers to engage with us on the project.
Prof. O'Brien and his team believe this has been an incredibly successful project in the short timeframe allotted and are very proud to have worked with the ERC in delivering this programme.
WP1: In vitro development and characterisation of miRNA-activated collagen (miRaColl) scaffolds
The primary outcome of this work package was the successful development and characterisation of functional miRNA-activated collagen (miRaColl) scaffolds. miRNA dose and delivery potential of various in-house developed delivery vectors including nanohydroxyapatite, polyethyleneimine and oligomeric chitosan were analysed in comparison to the commercial gold standard Lipofectamine RNAiMAX, and characterised using a combination of techniques in line with those optimised previously in Prof. O’Brien’s laboratory as part of a study on miRNA-activated scaffolds for bone repair. A particularly low dose of Lipofectamine RNAiMAX was optimised compared to doses previously reported in the literature, which resulted in the optimised Lipo-miRNA formulation being brought forward for further testing on our in-house developed collagen-hyaluronic acid scaffolds optimised specifically for cartilage-repair. miRNA levels were effectively manipulated across two microRNAs when assessed in MSC-seeded scaffolds. These specific targets were chosen based on the most recent evidence in the literature.
WP2: Testing and analysis of therapeutic miRaColl scaffolds in vitro using MSCs
Work undertaken in this theme demonstrated development of a novel therapeutic miRNA-activated scaffold but unsurprisingly, significant delays were experienced owing to technical issues encountered when optimising the system, due to this being a very nascent area of research. Firstly, this work focussed on testing functionality and therapeutic capacity of the miRNA-activated scaffolds containing miRNA-mimics, chosen based on the most recent evidence in the literature as indicated in WP1, on four human MSC donors. In parallel, we sought to identify additional chondrogenic miRNA targets of relevance within the three-dimensional environment of our in-house developed collagen-hyaluronic acid scaffolds due to the issues that arose while optimising the system. In order to do so, a high-throughput screening across 768 miRNAs was carried out identifying a remarkably novel miRNA. A comprehensive validation of the chondrogenic role of this novel miRNA is currently underway. Overall, the work undertaken in this theme demonstrated that miRaColl scaffolds enhanced expression of a range of cartilage-specific markers in comparison with the untreated controls. Cartilage matrix deposition analysis of miRaColl scaffolds revealed significant increases in sulphated glycosaminoglycan deposition, the hallmark constituent indicative of MSC chondrogenesis, when compared to relevant controls, pointing to the excellent potential of miRaColl to become a ground-breaking therapeutic with immense commercial interest.
WP3: Assessment of the miRaColl scaffold to regenerate full-thickness chondral defects in vivo
The work performed on this theme encountered delays due to technical difficulties incurred in WP2. However, as the optimal miRaColl system has now been identified, this promising miRaColl system has moved forward and in vivo work is currently ongoing; we are hopeful that a very positive result will be obtained in the coming months.
WP4: Commercial, regulatory and exploitation feasibility analysis of the miRaColl scaffold
The delays encountered in WP2 focusing on optimising the miRaColl system slowed down this commercialisation work package but since the successful miRaColl system has been identified and characterised, this work has proceeded to IDF submission and initial patentability analysis has confirmed that it is worthy of patent filing. As a whole, the thorough analysis conducted in WPs 1-3 has led to the development of a much more marketable technology that appears very promising. Relevant exploitation feasibility analysis is currently ongoing and some positive engagement with commercial partners has been conducted to-date which holds potential for the future.
Quantitative Output
To summarise some of the quantitative output from the project: 1 student completed a research Masters on the project (MSc in Bioengineering) and 1 student completed a summer research project. A number of publications are under preparation that are related to the project and will be published in leading peer-reviewed journals: 1 invited Progress Report focusing on “microRNA therapies in regenerative medicine” in Advanced Healthcare Materials; 1 invited Chapter on miRNA Modulation in Tissue Engineering and Regeneration: Cell Engineering and Regeneration.
Additionally, interactions with industry delegates at a number of events have resulted in positive engagement for progression of the commercialisation aspect of the project, including the 10th International MicroRNAs/Non-Coding RNAs & Genome Editing Europe 2015 Meeting held in Cambridge, UK, organised by GeneExpression Systems, Inc; the Controlled Release Society 2015 Annual Meeting held in Edinburgh, UK; the RNA Therapeutics - Advances and Opportunities 1st Innovation Workshop 2015 held in RCSI Dublin; the Gordon Research Conference on Musculoskeletal Biology and Bioengineering 2016 held in Andover, New Hampshire, US and the Exiqon microRNA & non-coding RNA Seminar 2016 held in Conway Institute UCD Dublin.
The research has been broadly covered by the media without compromising the commercialization element of the project and we have provided extensive opportunities for the next generation of researchers in the form of undergraduates, primary school and secondary school students and their teachers to engage with us on the project.
Prof. O'Brien and his team believe this has been an incredibly successful project in the short timeframe allotted and are very proud to have worked with the ERC in delivering this programme.