Periodic Reporting for period 1 - NANEMIAR (Nanomedicine Approach to Normalize Erythrocyte Maturation in Congenital Anemia by messenger RNA)
Período documentado: 2023-10-01 hasta 2025-03-31
The NANEMIAR project emerges amidst a transformative era in therapeutics, where conventional drug strategies have historically relied on small molecules. However, a paradigm shift is underway, with antibody-based drugs and genetic-based therapies for rare diseases gaining traction. This shift is driven by a growing recognition of the limitations of traditional approaches and the immense potential of innovative modalities to address previously unmet medical needs. In this context, NANEMIAR stands at the forefront of a therapeutic revolution, poised to harness the full potential of targeted delivery of therapeutic mRNA.
By fine-tuning mRNA expression and optimizing delivery mechanisms, NANEMIAR’s project ambition is to develop a therapy for congenital anemias such as β-thalassemia and Sickle Cell Disease. These conditions, characterized by disruptions in normal blood cell production, pose significant health burdens to affected individuals and their families. Current treatment options, while providing some relief, often fall short in efficacy and are associated with undesirable side effects. By leveraging mRNA as a therapeutic agent, NANEMIAR aims to overcome the challenges and effectively restore normal blood cell production while minimizing off-target effects. Within the 3-year project, we aim to deliver proof of concept of a bone marrow-targeted nanomedicine in human and animal thalassemia models. To this end, we defined the following key objectives (see project info sheet):
1. Development of a novel therapeutic mRNA with bone marrow specific delivery through targeted lipid nanoparticles (LNP). Activity will be analyzed in hematopoietic stem and progenitor cells (HSPCs) from healthy human donors and through (biodistribution) studies in mice.
2. Pharmaceutical testing in a thalassemic mouse model to determine dosing and toxicity of the therapeutic formulation.
3. Pre-clinical analysis of the therapeutic formulation in bone marrow cells from beta-thalassemic patients as proof of concept for efficacy.
By offering a safer, more cost-effective alternative to current treatment modalities, NANEMIAR's therapy has the potential to alleviate the burdens placed on healthcare systems and improve the quality of life for patients and their families. Additionally, by advancing our understanding of mRNA-based therapies and targeted delivery mechanisms, NANEMIAR contributes to the broader scientific and technological landscape, opening new avenues for innovation and discovery.
By fine-tuning mRNA expression and optimizing delivery mechanisms, NANEMIAR’s project ambition is to develop a therapy for congenital anemias such as β-thalassemia and Sickle Cell Disease. These conditions, characterized by disruptions in normal blood cell production, pose significant health burdens to affected individuals and their families. Current treatment options, while providing some relief, often fall short in efficacy and are associated with undesirable side effects. By leveraging mRNA as a therapeutic agent, NANEMIAR aims to overcome the challenges and effectively restore normal blood cell production while minimizing off-target effects. Within the 3-year project, we aim to deliver proof of concept of a bone marrow-targeted nanomedicine in human and animal thalassemia models. To this end, we defined the following key objectives (see project info sheet):
1. Development of a novel therapeutic mRNA with bone marrow specific delivery through targeted lipid nanoparticles (LNP). Activity will be analyzed in hematopoietic stem and progenitor cells (HSPCs) from healthy human donors and through (biodistribution) studies in mice.
2. Pharmaceutical testing in a thalassemic mouse model to determine dosing and toxicity of the therapeutic formulation.
3. Pre-clinical analysis of the therapeutic formulation in bone marrow cells from beta-thalassemic patients as proof of concept for efficacy.
By offering a safer, more cost-effective alternative to current treatment modalities, NANEMIAR's therapy has the potential to alleviate the burdens placed on healthcare systems and improve the quality of life for patients and their families. Additionally, by advancing our understanding of mRNA-based therapies and targeted delivery mechanisms, NANEMIAR contributes to the broader scientific and technological landscape, opening new avenues for innovation and discovery.
The first half of the project is mainly focused on the first objective of establishing the groundwork for a novel mRNA therapeutic that can be effectively delivered to specific cells in the bone marrow. We followed a parallel strategy on designing and testing mRNAs and LNPs in various models, including an immortalized cell model and primary HSPCs from human donors. Protocols for mRNA delivery have been validated across consortium partners and we demonstrated relevant levels of therapeutic protein expressed in HSPCs. Of note, LNP-mediated delivery appeared not straightforward in the HSPCs, so tests with various LNP formulations are ongoing. In addition, the physicochemical properties and shelf life of these new formulations are analysed to ultimately generate stable nanoparticles that can be used for targeted delivery.
Additionally, we have selected a series of targeting peptides that are able to bind a receptor present on specific cells in the bone marrow and confirmed receptor binding during a red blood cell differentiation assay of primary human HSPCs.
Preliminary data on LNP testing in mice indicates that we identified a novel LNP formulation that is able to avoid significant liver uptake (a phenomenon seen with most LNPs) and demonstrates uptake in bone marrow cells of the mice. This is a significant step forward and ideal starting point to investigate biodistribution and activity of targeted formulations in mice. For the targeted LNPs, we have established a robust conjugation technology to couple targeting moieties (e.g. peptides) to the LNP and are currently optimising this method to obtain a homogenous, stable and active targeted LNP formulation for follow-up experiments.
Additionally, we have selected a series of targeting peptides that are able to bind a receptor present on specific cells in the bone marrow and confirmed receptor binding during a red blood cell differentiation assay of primary human HSPCs.
Preliminary data on LNP testing in mice indicates that we identified a novel LNP formulation that is able to avoid significant liver uptake (a phenomenon seen with most LNPs) and demonstrates uptake in bone marrow cells of the mice. This is a significant step forward and ideal starting point to investigate biodistribution and activity of targeted formulations in mice. For the targeted LNPs, we have established a robust conjugation technology to couple targeting moieties (e.g. peptides) to the LNP and are currently optimising this method to obtain a homogenous, stable and active targeted LNP formulation for follow-up experiments.
The project aims to deliver significant scientific and technological advances by i) creating new knowledge on targeted non-viral delivery (via LNPs) of mRNA that may be used for other oligonucleotide strategies, such as siRNA or gene editing (e.g. CRISPR) therapies, and ii) demonstrating bone marrow targeting may be useful for therapy development for other rare hematological diseases. Finally, mRNA-based supplementation of gene defects can be used to de-risk future gene therapy approaches. Hence, outcomes in this project provide insights that are broadly applicable at three levels:
1. At the highest level, our study will indirectly affect up to 6% of the world population suffering from a genetic disease that is potentially curable by cell and gene therapy. The benefits range from quality-of-life improvements to many additional years of survival, depending on the disease.
2. At the mid-level, it offers a model therapeutic that can be exploited for other rare inherited anemias ineffective erythropoiesis.
3. And at the most immediate level, approximately 1.5% of the global population is β-thalassemia carrier, with an estimated incidence of symptomatic cases in the EU of 1 in 10.000 individuals.
Based on the above numbers, the NANEMIAR project has a clear scale-up potential and we are exploring potential pathways to commercialization by involving end-users and stakeholders in the project. When the therapeutic formulation shows promising results, the consortium can establish the tools and network to prioritize and perform the activities that are required to bring the formulation to the next phase: pharmacology, pharmacokinetics, and toxicology assessments, GMP setup, safety and efficacy studies in humans (Clinical Phase I/II). Ultimately, we aim for co-development with large pharma as they have the experience for Phase III studies and market approval.
1. At the highest level, our study will indirectly affect up to 6% of the world population suffering from a genetic disease that is potentially curable by cell and gene therapy. The benefits range from quality-of-life improvements to many additional years of survival, depending on the disease.
2. At the mid-level, it offers a model therapeutic that can be exploited for other rare inherited anemias ineffective erythropoiesis.
3. And at the most immediate level, approximately 1.5% of the global population is β-thalassemia carrier, with an estimated incidence of symptomatic cases in the EU of 1 in 10.000 individuals.
Based on the above numbers, the NANEMIAR project has a clear scale-up potential and we are exploring potential pathways to commercialization by involving end-users and stakeholders in the project. When the therapeutic formulation shows promising results, the consortium can establish the tools and network to prioritize and perform the activities that are required to bring the formulation to the next phase: pharmacology, pharmacokinetics, and toxicology assessments, GMP setup, safety and efficacy studies in humans (Clinical Phase I/II). Ultimately, we aim for co-development with large pharma as they have the experience for Phase III studies and market approval.