PROJECT TITLE:
Targeting OsteoArthritis with Senolytic and anti-Inflammatory peptide-loaded nanopharmaceuticals – OASIs
Project logo 
FOUNDING SOURCE: Programul 3 - Cooperare europeană și internațională
PROJECT CODE: ERANET-EURONANOMED-3-OASIs
CONTRACT NUMBER: 273/2022
PROJECT DURATION: February 2022 – December 2024
PROJECT BUDGET: RON 877.140 – 177.200 EURO
PROJECT CONSORTIUM:
PROJECT COORDINATOR - FUNDACIÓN INSTITUTO DE INVESTIGACIÓN SANITARIA DE SANTIAGO DE COMPOSTELA
SPAIN Department: Nano-Oncology and Translational Therapeutics Project coordinator: -Maria De La Fuentes
P2 -Fundación Profesor Novoa Santos –A Coruna Spain Project director:-Maria Mayan
P3 – State Research Institute Centre for Innovative Medicine Vilnius Lithuania Project director: Ali Mobasheri
P4-V-Nano Toulouse France Project director: - Frank Pawan
P5- NATIONAL INSTITUTE OF RESEARCH AND DEVELOPMENT FOR TECHNICAL PHYSICS (NIRDTP) Iasi Romania
Project director: - Luminita Labusca
SUMMARY
Osteoarthritis (OA) is the most common form of arthritis and a leading cause of pain and disability worldwide. Current treatments are limited to symptomatic relief. There is an urgent need to develop novel therapeutics that aim to restore joint function for the benefit of millions of people worldwide. Major limitations relate to the complexity of the underlying pathogenic mechanisms and molecular endotypes for the different clinical phenotypes, and to the need for efficient delivery strategies. OA treatment might be attained by personalizing care, accompanied by the development of targeted therapeutics that can successfully be delivered to the affected joint making use of nanotechnology.
OASIs aims to develop innovative nanopharmaceuticals based on the association of peptides that target senescence and inflammation to nanoemulsions that can safely be administered to the synovial joint via intra-articular injection, and provide sustained and long-lasting, symptom and structure modification, for the enhanced treatment of OA.
Interfering peptides targeting senescent cells can modulate tissue regeneration and response to injury. Anti-inflammatory peptides from biological venoms have shown to reduce inflammation and pain. In this proposal we will develop new combinatorial targeted therapies by loading these peptides into biocompatible and biodegradable, safe-by-design and versatile, lipid nanoemulsions. Combinations will be assessed in vitro, ex vivo and in vivo. Optimization of critical formulation parameters, imaging tracking and pharmacokinetic studies, will allow the selection of the most promising compositions for preclinical evaluation. The results obtained in this proposal will help us to make state-of-the-art advances in order to successfully advance our research to the clinic. Attention to critical manufacturing and regulatory processes, exploitation, dissemination and management activities will ensure translatability of the candidate nanopharmaceuticals.
OBJECTIVES
We aim to develop innovative nanoformulations that contain senolytic and anti-inflammatory peptides to target inflammation and senescence in ageing and inflammatory OA phenotypes, addressing an unmet clinical need by providing more efficient treatments to OA patients. These nanoformulations will also be helpful for the therapeutic management of other OA phenotypes where these two processes (senescence and inflammation) are involved in disease progression. Molecular understanding of the disease has allowed identifying new pathways that can be targeted with specific molecules such as peptides. Nanotechnology has the potential to allow local administration of combinatory treatments based on peptides and to provide a sustained release for a long-lasting therapeutic effect. We follow an innovative approach that merges pharmaceutical innovation, a deeper molecular understanding of disease pathogenesis, and personalization of the therapy.
The specific objectives are:
Objective 1. To develop nanoformulations that associate senolytic and anti-inflammatory peptides;
Objective 2. To optimize the technology for IA administration;
Objective 3. To study the best combinations in vitro and examine synergistic effects;
Objective 4. To determine the efficacy of the nanopharmaceuticals in animal models in inflammatory and ageing phenotypes of OA, generating a solid preclinical Proof of Concept (PoC); Objective 5. To scale-up and manufacture according to pharmaceutical regulations.
To achieve this ambitious aim, the consortium brings worldwide experts in OA, which can provide valuable expertise to define the most innovative therapeutic approaches as well as reliable models for generating a solid PoC, partners with expertise in drug discovery, nanomedicine and drug development, as well as industrial partners.
POTENTIAL CLINICAL IMPACT AND EXPECTED RESULTS
Clinical impact
OA is a leading cause of pain and disability internationally, affecting up to one third of the population over the age of 50 (15% of the European population, and more than 500 million people worldwide) 2, 31. The number of people affected by OA rose by 48% from 1990 till 2019 worldwide and is expected to increase due to ageing population and increased prevalence of obesity. While OA is a leading cause of disability in elderly, it is increasingly recognized that the global burden of disease is steadily increasing among people aged 15-49 years of age. When stratified by gender, there is a greater age-adjusted prevalence of OA in women (23.5%) as compared with men (18.1%) (32). In recent years, advances in innovative therapeutics have rather been limited, although the molecular understanding of the disease is providing new cues for the development of targeted therapeutics. OASIs aims to integrate new knowledge into a nanotechnology-based approach, following a patient-driven research, in order to provide new treatments that bring a therapeutic opportunity to OA patients. The preclinical results obtained in this proposal, accompanied by the delivery of manufacturing protocols and engagement of a manufacturer partner, the involvement of clinicians and relevant stakeholders, the definition of a product target profile and delivery of a regulatory roadmap, will advance the translation of the proposed research to the clinic.
Socio-economic impact
OA is a leading source of societal cost in older adults and significantly decreases quality of life. OA care costs more than Euro 4.8 million to treat in Spain alone (0.5 % GDP) (33). Most of the studies on economic costs have focused on the cost of arthroplasties and total joint replacement surgeries, but cost is likely to be far higher because of the lost working productivity associated with OA and health care costs. Secondary side effects of anti-inflammatory drugs in chronic patients, and comorbidities associated with OA are also impacting on quality of life and in health-economics. Prevention and disease modification are areas being targeted by various research endeavours, which have indicated great potential. The economic benefits of the development of disease-modifying therapies with a personalised approach proposed by OASIs have the potential to reduce the overall budget impact and trigger long-term improvements in quality of life for patients.
Expected Project results
This proposal is based on three patent applications. FIDIS, FNS and NIRDTP have full rights on the exploitations of these patents. Other innovations and protectable results are expected to come out of this project, which could generate new intellectual property for each partner itself and for the whole consortium. The EC, assisted by the Technology Transfer Offices of each participant, will follow the project to detect protectable results and decide the best ways of protection to maximize the entry of products into the market. In the long run we expect to obtain products for treatment of OA according to specific phenotypes based on nanotechnology. OASIs has engaged an industrial partner with capacity for GMP manufacturing, and clinical groups that can define the product target profile and establish a roadmap for entering clinical trials. Innovations are also expected in the field of diagnosis.
Potential users and uses of the results
The transfer of the results to the productive system could involve licensing IP, applying as a consortium for additional funds to continue with the development, as for example to the Eurostars programme or other funding opportunities that might arise in the EU's research and innovation funding programme 2021-2027, or incorporate a company specialized in personalized peptide therapies for OA. The strategy for the exploitation of results will be developed by the EC. To explore the options the consortium should investigate the market landscape for competitors. Pharmaceutical companies with focus in regenerative medicine, autoimmune and inflammatory diseases, will be approached. FIDIS is also setting-up a spin-off company, DIVERSA Technologies (www.diversatechnologies.com), co-founded by the coordinator of OASIs, specialized in providing drug delivery solutions with a focus on three therapeutic areas, cancer, inflammatory diseases and metabolic diseases, which could be interesting in licensing. All partners of OASIs have previous experience in tech transfer.
EVENTS
Kick – Off Consortium meeting
Santiago de Compostella Spain 3 May 2022
Dissemination
Conference presentation:
Tissue Engineering and Regenerative Medicine International Society - TERMIS European Chapter Conference 2022, 28.06.2022 - 1.07.2022, Krakow, Poland
A simplified protocol for preparation of cell based biological samples for observing nanomaterial surface adherence using scanning electron microscopy imaging (oral presentation)
Minuti A.E.; Lăbușcă L.; Herea D.D.; Chiriac H.; Lupu N.
Human mesenchymal stem cells and nanomagnetic materials for regenerative medicine (oral presentation)
Lăbușcă L.; Herea D.D.; Danceanu C.; Minuti A.E.; Stavila C.; Chiriac H.; Lupu N.
Magnetic Nanomaterials – an attractive opportunity for regenerative medicine – Luminita Labusca – podium presentation and session co-chair
Tissue Engineering and Regenerative Medicine International Society Asia-Pacific Chapter Conference 2022 - TERMIS AP, 5 – 8.10.2022, Jeju, South Korea
Establishing an attractive regenerative alliance: Human mesenchymal stem cells and nanomagnetic materials” (oral presentation)
Labusca L., Danceanu C., Minuti A., Herea D., Chiriac H., Lupu N.
International Cartilage repair and Joint presenrvation Society (ICRS), 12-16 Aprilie 2022 Berlin Germania
Human mesenchymal stem cells and nanomagnetic materials for regenerative, medicine (oral presentation)
Labusca L., Danceanu C., Minuti A., Herea D., Chiriac H., Lupu N.
Tissue Engineering and Regenerative Medicine International Society - TERMIS European Chapter Conference 2023, 27.03.2022 - 1.04.2023, Manchester, UK
Magnetoliposomes for intraarticular delivery of ascorbic acid and dexamethasone (poster presentation)
Zara Danceanu Camelia Mihaela .; Lăbușcă L.; Herea D.D.; Chiriac H.; Lupu N.
Human osteochondral explant tissue as reliable models for ex vivo testing of novel therapeutics in osteoarthritis (poster presentation)
Lăbușcă L.; Herea D.D.; Danceanu C.; Minuti A.E.; Stavila C.; Chiriac H.; Lupu N.
Conference CELLS AND TISSUES TRANSPLANTATION. ACTUALITIES AND PERSPECTIVES”
Dedicated to the 10th anniversary of the founding of the Human Tissue and Cells Bank and to the 15th anniversary of the founding of the Laboratory of Tissue Engineering and Cells Culture of Nicolae Testemitanu State University of Medicine and Pharmacy of the Republic of Moldova
Chisinau 17-18 March 2023
Session moderator- Luminita Labusca
Models for testing regenerative therapies – focus on explants as models for osteoarthritis. Iasi, Romania.
Labusca Luminita, Danceanu Camelia, Minuti Anca, Ivanov Iuliu, ZugunEloae Florin, Plamadeala Petru, Chiriac Horia, Lupu Nicoleta.
Reliable protocol for sample preparation to observe nanomaterial adherence to the surface of biological cells by using scanning electron microscopy. Iasi, Romania (oral presentation)
Stavila Cristina, Minuti A, Herea D, Labusca L, Stoian G, Lupu N, Chiriac H.
International cartilage repair and joint restauration (ICRS) 2023 World Congress Sitges Spain 9-12 September 2023
Osteochondral explant tissue -ex vivo models for testing novel therapies-
Labusca Luminita, Danceanu Camelia, Minuti Anca, Ivanov Iuliu, Zugun Eloae Florin, Plamadeala Petru, Chiriac Horia, Lupu Nicoleta (e-poster presentation)
Magneto-liposomes - platforms for intraarticular delivery of bioactive molecules
Zara Danceanu Camelia Mihaela .; Lăbușcă L.; Herea D.D.; Chiriac H.; Lupu N.
Conference: Regenerative medicine in orthopedic surgery (RMOS) IIIrd Global Summit Istanbul Turkey 30 Nov-2 Dec 2023
Magnetically enhanced adipose derived stem cell chondrogenesis
Luminita Labusca, Valentin Nastasa, Camelia Mihaela Zara Danceanu, Cristina Stavila, Anca Emanuela Minuti, Dumitru Daniel Herea, Razvan-Nicolae Malancus, Aurelian Sorin Pasca. Mihai Mares and Nicoleta Lupu. (poster presentation)
Other activities
Invited lecture/trainer Articular Cartilage Engineering Training School (ACETS) organized by Istinye University and COST action NetwOArk CA20111 Decembre 1-3 Istanbul Turkey -Luminita Labusca
Papers
- MAGNETIC NANOWIRES SUBSTRATE INCREASES ADIPOSE-DERIVED MESENCHYMAL CELLS OSTEOGENESIS
Labusca L., Danceanu C., Minuti A.E., Herea D.D., Ghemes A., Rotarescu C., Dragos-Pinzaru O., Tibu M., Grigoras M, Chiriac H. Lupu N.
Scientific Reports, vol. 12, art. no. 16698 (2022) - Herea, D., & Labusca, L. (2023). Magnetic Nanomaterials: Advanced Tools for Cartilage Extracellular Matrix Regeneration. Biomedical Research and Therapy, 10(1), 5509-5522. https://doi.org/10.15419/bmrat.v10i1.789
- Camelia-Mihaela Zară-Dănceanu, Anca-Emanuela Minuti, Cristina Stavilă, Luminiţa Lăbuscă, Dumitru-Daniel Herea, Crina Elena Tiron, Horia Chiriac, and Nicoleta LupuACS Omega 2023 8 (26), 23953-23963 DOI: 10.1021/acsomega.3c02449
- Zară-Dănceanu CM, Stavilă C, Minuti AE, Lăbușcă L, Nastasa V, Herea DD, Malancus RN, Ghercă D, Pasca SA, Chiriac H, Mares M, Lupu N. Magnetic Nanoemulsions for the Intra-Articular Delivery of Ascorbic Acid and Dexamethasone. Int J Mol Sci. 2023 Jul 25;24(15):11916. doi: 10.3390/ijms241511916. PMID: 37569290; PMCID: PMC10419142.
EXECUTIVE SUMMARY
In the framework of stage I "Optimization of manufacturing technologies of combined nanopharmaceuticals for intra-articular administration (WP2)" Fe3O4 magnetic nanoparticles covered with lipids(oleic acid and palmitic acid) were prepared by methods that produced homogeneous and reproducible particles, a protocol that lends itself to being adapted to industrial production. The particles were characterized from a physico-chemical point of view (dimensions, magnetization, composition) using advanced equipment existing within INCDFT-IFT, Iasi. The characterization was also carried out in terms of interactions with relevant in vitro models, at the cellular level (mesenchymal stem cells from adipose tissue and human fibroblasts) in terms of viability, membrane integrity and the expression of senescence markers. An excellent viability and even proliferation was noted with increasing doses of nanoparticles, as well as the preservation of membrane integrity and the decrease of cellular senescence markers.
Nanoemulsions were prepared and their coupling with magnetic nanoparticles was achieved through an original protocol using a mixed method of low and high energy (short-term ultrasonication).
A physical characterization of the complexes of nanoparticles - lipid emulsions with active substances with an anti-aging and anti-inflammatory role (ascorbic acid and dexamethasone) was made, finding the submicron size and the excellent stability in the emulsion in the short and medium term, the observations continue for the long-term follow-up long.
The obtained results allow the assessment that it is possible to proceed to the second part of the characterization stage by continuing the testing of the complexes on an in vitro cell model and later on an organ explant consisting of osteocartilaginous and synovial tissue with the evaluation of markers of senescence, inflammation and tissue regeneration.
During OASIS stage II titled “Optimization of nano pharmaceutical formulations for intra-articular administration (WP2) " the testing of complex nanomagnetic particles - nano emulsions loaded with pharmacological substances on an in vitro cell model continued and their role in increasing the mobility and targeting capacity of stem cells was highlighted like the anti-senescent effect. An organ explant model of arthrosis using fragments of osseocartilaginous tissue and, for the first time for human tissues, in co-culture with synovial tissue from the same joint was achieved. The ability of this system to reproduce in vitro the complex interactions existing at the level of joints affected by arthrosis and the ability to respond to stimuli with therapeutic potential has been hereby demonstrated. The results we obtained justify the use of the mixed osteochondral tissue-synovial tissue explant method as a way of ex vivo testing of new therapeutics for arthrosis. The effect of magnetic Nano emulsions loaded with pharmaceutical substances was also tested on an in vivo model in mice and it was found that they do not exert an inflammatory or toxic effect after systemic injection (intraperitoneal delivery). The obtained results were disseminated to scientific audience by presentations at international conferences and published in specialized scientific journals indexed in Web of Science.