Leveraging RNA nanotechnology for next-generation gene delivery systems
Lead Research Organisation:
Sixfold Bioscience Ltd
Department Name: Head office
Abstract
Can we built efficient gene delivery systems to target the brain? Gene therapy offers the promise to effectively revert the cause of inherited and acquired brain diseases. However, the brain is arguably the most challenging organ to target with therapeutic agents, especially large-size gene therapeutics. This is due to the existence of the blood brain barrier (BBB) - a highly selective border separating the blood circulation from the brain, formed by a continuous layer of sealed endothelial cells embedded in the capillary basement membrane. The BBB physiologically protects the neurons from neurotoxic factors present in the bloodstream, but also prevents exogenously administered medications to reach their target of action. Non-targeting brain therapies could not only result in poor efficacy of the treatment, but also in side effects due to the accumulation of toxic drugs in other organs, encumbering the life of the patient. Lipid nanoparticles were thought to cross the BBB, thanks to their charge and size by abortpion mediated endocytosis or transcytosis. But this approach failed to provide efficient delivery, testified by the lack of approved gene therapies targeting the brain.
Can we use nucleic acids as gene delivery systems to target the brain? Nucleic acids are unstable natural polymers that degrade by nucleases upon intravenous injection and thus far were considered unsuitable for use as drug delivery vehicles. Yet, they have inherent properties to accommodate spatially arranged molecules and controlled stoichiometry for precise shape and size nanoparticles, crucial for an efficient tissue and cell targeting. The new approach involves the introduction of nuclease stable RNA nanoparticles, based on Sixfold's Ltd patent portfolio for the exploitation of DNA and RNA-based nanoparticles and targeting molecules including, but not limited to, aptamers, for the delivery of selected gene therapeutics. In preliminary studies, these particles have shown to accumulate in the brain.
The research programme focuses on the assessment of the causal relationship between RNA nanoparticles charge, lipophilicity and size; to tissue/cell targeting. The nature of repetitive units (nucleotides) of nucleic acids, allows for precise design of the delivery systems in silico and prediction of the nanoparticle behaviour using Machine Learning. This offers a high degree of flexibility to create an on-demand platform for tissue targeting and reduce the physical experimentation. A toolbox of methods allowing for the nanoparticles stoichiometric characterization and biodistribution monitoring imaging will be developed. Preclinical demonstrators of the platform will be assessed thanks to the engagement of pioneering research groups working on animal models on neurodegenerative diseases including Parkinson's and Kennedy's (spinal bulbar muscular atrophy) diseases.
Can we use nucleic acids as gene delivery systems to target the brain? Nucleic acids are unstable natural polymers that degrade by nucleases upon intravenous injection and thus far were considered unsuitable for use as drug delivery vehicles. Yet, they have inherent properties to accommodate spatially arranged molecules and controlled stoichiometry for precise shape and size nanoparticles, crucial for an efficient tissue and cell targeting. The new approach involves the introduction of nuclease stable RNA nanoparticles, based on Sixfold's Ltd patent portfolio for the exploitation of DNA and RNA-based nanoparticles and targeting molecules including, but not limited to, aptamers, for the delivery of selected gene therapeutics. In preliminary studies, these particles have shown to accumulate in the brain.
The research programme focuses on the assessment of the causal relationship between RNA nanoparticles charge, lipophilicity and size; to tissue/cell targeting. The nature of repetitive units (nucleotides) of nucleic acids, allows for precise design of the delivery systems in silico and prediction of the nanoparticle behaviour using Machine Learning. This offers a high degree of flexibility to create an on-demand platform for tissue targeting and reduce the physical experimentation. A toolbox of methods allowing for the nanoparticles stoichiometric characterization and biodistribution monitoring imaging will be developed. Preclinical demonstrators of the platform will be assessed thanks to the engagement of pioneering research groups working on animal models on neurodegenerative diseases including Parkinson's and Kennedy's (spinal bulbar muscular atrophy) diseases.
Planned Impact
The project will have wide-ranging Academic, Economic & Societal impacts (no significant Environmental).
Academic
-Advances in understanding and methods:The project will elucidate the processes involved in the control over cell and tissue targeting via nucleic acid delivery systems, which remain not very well understood. To propagate the knowledge and methods, we estimate that the project will lead to two presentations at academic conferences per year (e.g.Gordon Research Conference in RNA Technology), and 2-3 publications (e.g.Nucleic Acids Research), subject to Intellectual Property considerations.
-Knowledge transfer:The project involves 3 academic collaborations (with Prof M Wood at Oxford University, Dr D Stuckley at UCL and Prof L Alvarez-Erviti at CBIR, Spain), who will provide their expertise and model systems to elevate the project and advise Dr Mylonaki. The project will help develop Dr Mylonaki's and her team's knowledge.To further leverage the wider academic community, once per year Dr Mylonaki will organise an on-site mini-symposium to discuss potential broader application of the research.
Economic
-Proof-of-concept data/New products:The project will generate data on incorporating Dr Mylonaki's novel strategies into Sixfold's PODS for optimised therapeutic safety and efficacy in the proof-of-concept CNS indications, where there are large unmet needs. These data will supplement Sixfold's preclinical data pack for commercialisation of innovation.We will investigate the feasibility of turning Dr Mylonaki's innovations into separate commercial products to be licensed for use in therapies other than PODS. These activities will likely lead to new IP, strenghtening Sixfold's business proposition.
-Increased revenue from licensing:Sixfold's business model is based on licensing of preclinical PODS assets for further clinical development to pharma/biotech companies. We estimate the direct benefit from Dr Mylonaki's project to have return on investment (ROI) >817% by 2025. This is driven by accelerated completion of preclinical testing with increased competitiveness due to higher efficacy aand pipeline extension.
-Establishment of Sixfold's CNS Department and Job Creation: Dr Mylonaki and her project will contribute to the establishment and expansion of Sixfold's CNS department. Importantly, the project will directly lead to creation of 3FTEs, and indirectly will result in estimated additional c.20FTEs by 2025.
-Knowledge transfer to and from industry players:The project will engage multiple industry players (incl. mentors from GSK & Autifony) to create a bi-directional feedback on the products, unmet needs and best industry and commercialisation practices to maximise value creation in a sustainable fashion.
-UK's economic competitiveness:With the research positioned at an innovative SME, the project will involve speedy and effective research and commercialisation activities contributing to the enhancement of UK's competitiveness in the emerging field of nucleic acid therapeutics and ATMPs.
Societal
-Outreach activities:We will work with various universities to give talks and presentations to children and students at various levels of education on the importance and benefits of Advanced Therapies and working in innovative start-ups/SMEs. We will also advocate for ATMPs adoption.
-Enhancing quality of life and health for patients (esp. neurodegenerative):Dr Mylonaki's research has the potential to increase the safety and efficacy of Sixfold's and other companies' therapeutics, with the ultimate aim of contributing to the expedited creation of better nuclei acid-based therapies for patients worldwide.E.g. UK:145k people are living with Parkinson's (2018) acc. to the CPRD, at the cost of £449mn-£3.3bn annually.
-Meeting Government priorities:The project is timely given favourable regulatory environment and addresses the UK's Grand Challenge related to the Aging Society.The project is well aligned with IUK
Academic
-Advances in understanding and methods:The project will elucidate the processes involved in the control over cell and tissue targeting via nucleic acid delivery systems, which remain not very well understood. To propagate the knowledge and methods, we estimate that the project will lead to two presentations at academic conferences per year (e.g.Gordon Research Conference in RNA Technology), and 2-3 publications (e.g.Nucleic Acids Research), subject to Intellectual Property considerations.
-Knowledge transfer:The project involves 3 academic collaborations (with Prof M Wood at Oxford University, Dr D Stuckley at UCL and Prof L Alvarez-Erviti at CBIR, Spain), who will provide their expertise and model systems to elevate the project and advise Dr Mylonaki. The project will help develop Dr Mylonaki's and her team's knowledge.To further leverage the wider academic community, once per year Dr Mylonaki will organise an on-site mini-symposium to discuss potential broader application of the research.
Economic
-Proof-of-concept data/New products:The project will generate data on incorporating Dr Mylonaki's novel strategies into Sixfold's PODS for optimised therapeutic safety and efficacy in the proof-of-concept CNS indications, where there are large unmet needs. These data will supplement Sixfold's preclinical data pack for commercialisation of innovation.We will investigate the feasibility of turning Dr Mylonaki's innovations into separate commercial products to be licensed for use in therapies other than PODS. These activities will likely lead to new IP, strenghtening Sixfold's business proposition.
-Increased revenue from licensing:Sixfold's business model is based on licensing of preclinical PODS assets for further clinical development to pharma/biotech companies. We estimate the direct benefit from Dr Mylonaki's project to have return on investment (ROI) >817% by 2025. This is driven by accelerated completion of preclinical testing with increased competitiveness due to higher efficacy aand pipeline extension.
-Establishment of Sixfold's CNS Department and Job Creation: Dr Mylonaki and her project will contribute to the establishment and expansion of Sixfold's CNS department. Importantly, the project will directly lead to creation of 3FTEs, and indirectly will result in estimated additional c.20FTEs by 2025.
-Knowledge transfer to and from industry players:The project will engage multiple industry players (incl. mentors from GSK & Autifony) to create a bi-directional feedback on the products, unmet needs and best industry and commercialisation practices to maximise value creation in a sustainable fashion.
-UK's economic competitiveness:With the research positioned at an innovative SME, the project will involve speedy and effective research and commercialisation activities contributing to the enhancement of UK's competitiveness in the emerging field of nucleic acid therapeutics and ATMPs.
Societal
-Outreach activities:We will work with various universities to give talks and presentations to children and students at various levels of education on the importance and benefits of Advanced Therapies and working in innovative start-ups/SMEs. We will also advocate for ATMPs adoption.
-Enhancing quality of life and health for patients (esp. neurodegenerative):Dr Mylonaki's research has the potential to increase the safety and efficacy of Sixfold's and other companies' therapeutics, with the ultimate aim of contributing to the expedited creation of better nuclei acid-based therapies for patients worldwide.E.g. UK:145k people are living with Parkinson's (2018) acc. to the CPRD, at the cost of £449mn-£3.3bn annually.
-Meeting Government priorities:The project is timely given favourable regulatory environment and addresses the UK's Grand Challenge related to the Aging Society.The project is well aligned with IUK
Publications
Sixfold Bioscience
(2023)
A revolution in targeted RNA delivery
in Nature Research, advertisement feature
Description | Data generated during Year 3 are supporting Sixfold's ongoing Series A funding round and attracted key opinion leaders to create Sixfold's Strategic Advisory Board. Data obtained during Year 1-2 supported Sixfold's successful seed round of £7.8M. A provisional patent was submitted under the number 63/175,185. Although rapidly expanding in recent years, the RNA therapeutics field is still largely constrained to liver-targeting/local administration. This award has contributed to the development of Sixfold's Mergo® platform, holding potential to expand this promising therapeutic approach to other organs for a range of therapeutic indications. Mergo® is a chemically modified oligonucleotide tag for cell- or tissue-specific delivery of RNA oligonucleotides. The building-blocks that form the Mergo tag are computationally screened, assembled using high-throughput chemistry, and tested, with each iteration feeding into the next (Objective A). Cell specific-targeting was also assessed, resulting in different Mergo chemistries being uptaken by specific cell types (Objective B). A therapeutic indication relevant to the cell types uptaking Mergo was identified (Objective C). |
Exploitation Route | Sixfold will leverage the data generated under this award for continued exploration of partnership opportunities with selected pharma partners (currently in discussions with multiple large pharma companies for pilot collaborations, partially based on data outputs from this project). Data from this project contributes to larger data packages shared by Sixfold with potential partners interested in collaboration opportunities. Sixfold will also leverage the data to apply for further public funding (including one follow-on BMC grant application has been submitted and granted). |
Sectors | Pharmaceuticals and Medical Biotechnology |
URL | https://www.sixfold.bio/platform |
Description | The grant funding has enabled the publication of a patent submitted during Year 1 (under no. 63/175,185, PCT/IB2022/000218), and an additional patent submission in Year 2 (63/412,070). Project outputs have led to further funding, including a second IAA with University of Sheffield. The project has generated significant economic impacts on private sectors via utilisation of external facilities and resources, including access to analytical capabilities: mass-spectrometry facility at the Centre for Rapid Online Analysis of Reactions (ROAR), qPCR at Synbicite, as well as Royce at Imperial facilities. Academic impacts include collaborations with Dr Alice Pyne, Research Fellow in Biophysics at the London Centre for Nanotechnology to enable the high-resolution images of Mergo candidates. Year 2 has continued our strong partnership with CRO Pharmidex which has provided preliminary in vivo data to guide our Year 3 objectives. This project has contributed key datasets to the generation of Sixfold's virtual due-diligence data room (to secure Series A funding) in Year 3. |
Sector | Pharmaceuticals and Medical Biotechnology |
Impact Types | Societal Economic |
Description | Biomedical Catalyst |
Amount | £889,213 (GBP) |
Funding ID | 10026765 |
Organisation | Innovate UK |
Sector | Public |
Country | United Kingdom |
Start | 03/2022 |
End | 03/2024 |
Description | Biomedical Catalyst |
Amount | £499,833 (GBP) |
Funding ID | 10072572 |
Organisation | Innovate UK |
Sector | Public |
Country | United Kingdom |
Start | 11/2023 |
End | 10/2025 |
Description | Impact Acceleration Account |
Amount | £35,597 (GBP) |
Organisation | Engineering and Physical Sciences Research Council (EPSRC) |
Sector | Public |
Country | United Kingdom |
Start | 02/2022 |
End | 06/2022 |
Description | Leap R3 RNA Readiness + Response |
Amount | $1,493,227 (USD) |
Organisation | Wellcome LEAP |
Sector | Charity/Non Profit |
Country | United States |
Start | 01/2022 |
End | 12/2024 |
Description | RTO Scheme |
Amount | £15,000 (GBP) |
Organisation | Innovate UK |
Sector | Public |
Country | United Kingdom |
Start | 01/2023 |
End | 03/2023 |
Description | Royce Industrial Collaboration Program |
Amount | £82,045 (GBP) |
Funding ID | RICP21-0007 |
Organisation | Henry Royce Institute |
Sector | Academic/University |
Country | United Kingdom |
Start | 03/2021 |
End | 09/2021 |
Description | High resolution AFM of nanostructures |
Organisation | Henry Royce Institute |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | My team conceives, develops, produces, and provides the material for AFM analysis, consisting of RNA nanostructures. |
Collaborator Contribution | The team of Dr Pyne images the RNA nanostructures at their state-of-the-art AFM facility. Dr Pyne's team intellectual input and expertise were crucial for the sample preparation, resulting in successful imaging. They also design a platform for the high-throughput analysis of the images. |
Impact | This collaboration resulted in the high-resolution imaging of RNA nanostructures in an aqueous environment. This allows us to assess the conformation of the RNA nanostructures which is useful as a quality control tool. The quality of the images was so advanced, that allows for observing the double helix structure. This is a multidisciplinary collaboration between Dr Pyne's team of physicists and my team of chemists and biochemists. |
Start Year | 2021 |
Description | In vivo work in collaboration with Pharmidex |
Organisation | Pharmidex |
Country | United Kingdom |
Sector | Private |
PI Contribution | This collaboration allows Pharmidex to develop a more comprehensive suite of in vivo experimentation services including an expansion into oligonucleotide-based therapeutics, benefiting from my team's expertise in the expanding field of oligonucleotides |
Collaborator Contribution | Pharmidex performed the in vivo experimentation necessary for this project, leveraging their animal facility and in vivo expertise. |
Impact | The in vivo experiments conducted with Pharmidex enabled us to assess the biodistribution of RNA nanostructures in tissues, and accordingly select diseases of interest. |
Start Year | 2021 |
Description | Studying sub-cellular distribution of Sixfold's Mergo platform using a humanised model |
Organisation | Medicines Discovery Catapult |
Country | United Kingdom |
Sector | Private |
PI Contribution | Different Mergo chemistries were screened in vivo at Sixfold Bioscience. The top two promising candidates were produced and fluorescently labelled for this collaboration. |
Collaborator Contribution | Spheroids were assembled by MDC using primary human cells. The spheroids were incubated in the presence of Mergos. Microscopy was used to assess cell specific uptake. |
Impact | This collaboration allowed us to validate cell specific uptake in humans cells. This reinforces the translation of our previous findings in animals towards clinical application in humans. This is a multidisciplinary collaboration including the cell biologists that develop spheroids (MDC) with expertise in confocal microscopy, as well as chemists developing the Mergo drug delivery system (Sixfold Bio). |
Start Year | 2022 |
Title | COMPOSITIONS CONTAINING NUCLEIC ACID NANOPARTICLES WITH MODULAR FUNCTIONALITY |
Description | The invention provides compositions containing cargo molecules attached to elements that improve the function of the cargo molecules in the body of a subject. The compositions are useful for therapeutic and diagnostic purposes. Furthermore, the invention outlines ways in which these compositions can be produced; the core molecule can be functionalized, via bioorthogonal click chemistry, in such a way as to impart modular characteristics. This functionalization simultaneously allows for loading of biologically relevant cargo and provides stabilization to the overall structure of the molecule. |
IP Reference | US2022370635 |
Protection | Patent / Patent application |
Year Protection Granted | 2022 |
Licensed | Commercial In Confidence |
Impact | abc |
Description | Bioscience student outreach project |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Undergraduate students |
Results and Impact | Sixfold hosted 2 students from the University of Nottingham, as part of a summer project addressing the awarding gap in R&D careers/higher education. The students shadowed a member of the team (Lara), where they were given insights into experimental lab techniques and information on post-project impacts. The students also had the opportunity to talk with several members of the team to expand their professional network and gain insight on research interests and different roles in biotech. |
Year(s) Of Engagement Activity | 2022 |
URL | https://www.linkedin.com/posts/sixfoldbioscience_stem-awardinggap-activity-6984149596608339969-TY-j?... |
Description | Interview by the UK Bioindustry Association: Stepping into leadership roles with UKRI's Future Leaders Fellowships. |
Form Of Engagement Activity | Engagement focused website, blog or social media channel |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Industry/Business |
Results and Impact | I was interviewed by the UK Bioindustry Association to increase awareness of how the UKRI FLF enables talent to step into leadership roles in biotech. Following this, potential FLF applicants reached out to me requesting information on how to shape their application and the application process for business-hosted applicants. |
Year(s) Of Engagement Activity | 2023 |
URL | https://www.bioindustry.org/news-listing/stepping-into-leadership-roles-with-ukris-future-leaders-fe... |