Periodic Reporting for period 2 - CARTHAGO (Cartilaginous tissue regeneration by non-viral gene therapy; taking the hurdles towards efficient delivery)
Okres sprawozdawczy: 2022-09-01 do 2024-08-31
Diseases of the musculoskeletal system impose a substantial burden on Western societies, which is ever increasing with ageing of the population. Amongst the diseases with most impact are osteoarthritis (OA) and chronic low back pain (CLBP) caused by intervertebral disc (IVD) degeneration, involving the cartilaginous tissues of these organs, being main causes of loss of disability-adjusted life years. Moreover, absenteeism poses a substantial economic burden on society. Despite the severity of the problems, medical solutions currently are limited and consist mainly of highly invasive surgery that replaces or immobilises the joint. Significant effort worldwide is aimed at helping the joint and IVD heal themselves by regenerative medicine, using stem cells or growth factors to regenerate the tissue, yet very few of these strategies have entered the clinic. Non-viral gene therapy has the potential to overcome these hurdles, due to its high specificity and, hence, safety, longevity of effects, and cost-effectiveness. However, the tight extracellular matrix of cartilaginous tissue does not contain blood vessels for transport and distribution of the therapeutic nucleic acids, posing a challenge to the delivery of gene activity modifying agents, in addition to prevention of extracellular and intracellular degradation of the therapeutic nucleic acids. Furthermore, in the public opinion gene therapy is conceived as a potentially hazardous strategy, and awareness and adequate information of the public is critical to enhance well-informed adoption.
In CARTHAGO, together with 15 Early Stage Researchers (ESRs) we have addressed these challenges through an integrated research and training programme focused on 1) cell delivery and efficiency of gene modulation, 2) tissue and organ delivery tools; 3) repair in tissue and organ culture; 4) in vivo imaging of regeneration and gene therapy efficacy; and 5) responsible research and innovation. We have generated novel insights as well as new tools and concepts that will not only advance research in the field of gene therapy for cartilaginous tissue regeneration, but will also be of great interest to other scientific disciplines including nucleic acid therapy, biomaterials, gene delivery, regenerative medicine, orthopaedics in general, and research ethics. This will benefit the development of better treatment of musculoskeletal disorders as well as of other diseases eligible for nonviral gene therapy, contributing to the future accessibility of safe and effective therapy to patients and healthcare professionals. In addition, our project has delivered a next generation of young scientists trained to tackle the current challenges of gene therapy who can become our future scientific leaders in relevant fields.
In CARTHAGO, together with 15 Early Stage Researchers (ESRs) we have addressed these challenges through an integrated research and training programme focused on 1) cell delivery and efficiency of gene modulation, 2) tissue and organ delivery tools; 3) repair in tissue and organ culture; 4) in vivo imaging of regeneration and gene therapy efficacy; and 5) responsible research and innovation. We have generated novel insights as well as new tools and concepts that will not only advance research in the field of gene therapy for cartilaginous tissue regeneration, but will also be of great interest to other scientific disciplines including nucleic acid therapy, biomaterials, gene delivery, regenerative medicine, orthopaedics in general, and research ethics. This will benefit the development of better treatment of musculoskeletal disorders as well as of other diseases eligible for nonviral gene therapy, contributing to the future accessibility of safe and effective therapy to patients and healthcare professionals. In addition, our project has delivered a next generation of young scientists trained to tackle the current challenges of gene therapy who can become our future scientific leaders in relevant fields.
Our 15 ESRs have conducted their research in 5 Work Packages (WPs) to address CARTHAGO’s objectives. Their main work and results are summarised below:
In WP2, we have developed polymeric nanoparticles, dendrimers, albumin biomolecular assemblies, microbubbles and hydrogels and have demonstrated that these can successfully enter cells and change their expression profile. The challenge of entry and visualisation of nucleic acids in the complex native tissues of the joints and IVD have been further demonstrated by us, with the nanoparticles showing entry into the dense tissues of the joint. Statistical approaches can support the efficient design of nanoparticles, dendrimer and hydrogel-based nucleic acid therapeutics formulations and, hence, treatment development.
In WP3, we have implemented non-linear ultrasound technology and have demonstrated that non-linear ultrasound is a promising novel tool to enhance tissue permeation of nucleic acids, either naked or within carrier systems, in cartilage tissue and to locally trigger dissociation and release of albumin-bound oligonucleotides upon systemic administration.
In WP4, we have obtained preliminary proof of concept for the ability of antisense oligos to modify gene expression delivered systemically within albumin biomolecular assemblies. This approach is promising for local treatment either or not with ultrasound for treatment ‘on demand’. Furthermore, cartilage targeting components can be built into albumin carrier design.
In WP5, we have demonstrated ex vivo proof-of-concept for click chemistry is as a useful tool to specifically demonstrate cartilage and IVD matrix regeneration in existing tissue. Our in vivo studies in a rat model of ACLT-DMM-induced OA showed that high affinity FcRn binding albumin biomolecular assemblies to enhance the circulatory half-life of antisense therapeutic oligonucleotides is safe for intra-articular injection. These promising results warrant further research to explore the impact on pain and inflammation.
In WP6, we developed and published a draft for the first educational framework incorporating ethics in biomedical development. This is a useful concept for responsible research and innovation in biomedical research.
We have shared our results via 19 scientific publications up to date, presentations at conferences, including two dedicated symposia, and on our website, X and LinkedIn for lay audience including patients. Also a local outreach event was organised in Davos for the local population.
In WP2, we have developed polymeric nanoparticles, dendrimers, albumin biomolecular assemblies, microbubbles and hydrogels and have demonstrated that these can successfully enter cells and change their expression profile. The challenge of entry and visualisation of nucleic acids in the complex native tissues of the joints and IVD have been further demonstrated by us, with the nanoparticles showing entry into the dense tissues of the joint. Statistical approaches can support the efficient design of nanoparticles, dendrimer and hydrogel-based nucleic acid therapeutics formulations and, hence, treatment development.
In WP3, we have implemented non-linear ultrasound technology and have demonstrated that non-linear ultrasound is a promising novel tool to enhance tissue permeation of nucleic acids, either naked or within carrier systems, in cartilage tissue and to locally trigger dissociation and release of albumin-bound oligonucleotides upon systemic administration.
In WP4, we have obtained preliminary proof of concept for the ability of antisense oligos to modify gene expression delivered systemically within albumin biomolecular assemblies. This approach is promising for local treatment either or not with ultrasound for treatment ‘on demand’. Furthermore, cartilage targeting components can be built into albumin carrier design.
In WP5, we have demonstrated ex vivo proof-of-concept for click chemistry is as a useful tool to specifically demonstrate cartilage and IVD matrix regeneration in existing tissue. Our in vivo studies in a rat model of ACLT-DMM-induced OA showed that high affinity FcRn binding albumin biomolecular assemblies to enhance the circulatory half-life of antisense therapeutic oligonucleotides is safe for intra-articular injection. These promising results warrant further research to explore the impact on pain and inflammation.
In WP6, we developed and published a draft for the first educational framework incorporating ethics in biomedical development. This is a useful concept for responsible research and innovation in biomedical research.
We have shared our results via 19 scientific publications up to date, presentations at conferences, including two dedicated symposia, and on our website, X and LinkedIn for lay audience including patients. Also a local outreach event was organised in Davos for the local population.
Through peer reviewed publications in scientific journals and open access availability of the generated data, CARTHAGO has strengthened overall European innovativeness. For the participating industrial partners, access to scientific excellence and novel insights into the applicability of their technology was facilitated to follow up upon. This is significant as health, demographic change and well-being is a challenge of top priority in Europe. On a wider scale, many achievements of CARTHAGO in the fields of nucleic acid therapy, biomaterials, gene delivery, regenerative medicine, orthopaedics in general, and research ethics reach far beyond their application in cartilage and intervertebral disc degeneration and can be translated to the treatment of other diseases.
For the society, the general population, the patients, health care professionals and health care systems, CARTHAGO contributes to the investigation of the application of nucleic acid therapy as safe and effective treatment approach. Nucleic acid technology holds great promise for treatment of musculoskeletal disorders, and follow-on studies are warranted. Ethical aspects were assessed in parallel to the research activities throughout CARTHAGO, assuring the consideration of potential concerns. Moreover, the ethical research in CARTHAGO laid the foundation for integration of ethics research in biomedical science in general, thereby allowing the expansion of ethical awareness throughout the European scientific community. Through public engagement and training of ESRs in public science communication CARTHAGO also has impact on public awareness of the potential of future therapies and thereby their adoption by patients and caregivers. Given that osteoarthritis and back pain are top causes for disability on a global scale, the economic impact of the availability of sustainable therapies is substantial.
For the society, the general population, the patients, health care professionals and health care systems, CARTHAGO contributes to the investigation of the application of nucleic acid therapy as safe and effective treatment approach. Nucleic acid technology holds great promise for treatment of musculoskeletal disorders, and follow-on studies are warranted. Ethical aspects were assessed in parallel to the research activities throughout CARTHAGO, assuring the consideration of potential concerns. Moreover, the ethical research in CARTHAGO laid the foundation for integration of ethics research in biomedical science in general, thereby allowing the expansion of ethical awareness throughout the European scientific community. Through public engagement and training of ESRs in public science communication CARTHAGO also has impact on public awareness of the potential of future therapies and thereby their adoption by patients and caregivers. Given that osteoarthritis and back pain are top causes for disability on a global scale, the economic impact of the availability of sustainable therapies is substantial.