Periodic Reporting for period 1 - NanoImmunoRT (Synergizing enhanced radiotherapy with tumor-associated macrophages-targeted immunotherapy in glioblastoma with multimodal therapeutic nanoparticles)
Reporting period: 2024-01-01 to 2025-12-31
Summary of the context and overall objectives of the project
Glioblastoma (GBM) is the most aggressive and common form of primary brain cancer in adults. Despite advances in surgery, radiotherapy (RT), and chemotherapy, the prognosis for GBM patients remains poor, with median survival rarely exceeding 15 months. Two major challenges limit the efficacy of current treatments. The immunosuppressive tumor microenvironment, particularly the presence of tumor-associated macrophages (TAMs), which promote tumor growth and suppress anti-tumor immune responses, and hypoxia (low oxygen levels) within the tumor which further exacerbates these issues, reducing the effectiveness of RT and immunotherapy. Addressing these limitations requires innovative, multimodal therapeutic strategies that can simultaneously enhance RT efficacy, reprogram the tumor immune microenvironment, and overcome hypoxia.
Project objectives
The NanoImmunoRT project aims to develop a synergistic therapeutic strategy combining radiosensitization and TAM-targeted immunotherapy using multimodal therapeutic nanoparticles (MTNPs). The project leverages zeolite-based nanoparticles (LTL types) functionalized with manganese (Mn) and gadolinium (Gd) to enhance catalytic activity for oxygen generation and radiosensitization, and toll-like receptor (TLR) agonists (e.g. R848) to reprogram TAMs and stimulate anti-tumor immune responses.
The overall goal is to create a single nanoparticle platform that can:
- Decompose tumoral hydrogen peroxide (H2O2) into oxygen (O2), alleviating hypoxia and improving RT efficacy.
- Act as a radiosensitizer, enhancing DNA damage in cancer cells during RT.
- Re-educate TAMs from a pro-tumor (M2) to an anti-tumor (M1) phenotype, boosting the immune response against the tumor.
Project Pathway to Impact
The project introduces a novel MTNP formulation (LTL-Mn/Gd nanozeolites) with superior catalytic activity and radiosensitizing properties, addressing key limitations of current GBM therapies. In vitro and in vivo studies demonstrated that LTL-Mn/Gd nanozeolites enhance RT efficacy, reduce tumor hypoxia, and synergize with immunotherapy (R848), leading to significant tumor growth inhibition and prolonged survival in murine models. The project provides a proof-of-concept for combining RT and immunotherapy using MTNPs, paving the way for future preclinical studies in GBM and other hypoxic, immune-suppressive tumors.
Scale and significance of expected impacts
The project’s findings could help reshape the field of radioimmunotherapy, demonstrating the feasibility of using MTNPs to overcome hypoxia and immune suppression in GBM. The development of patentable MTNPs could stimulate innovation in the biotech sector, creating new markets and job opportunities. Moreover, by engaging with patients, clinicians, and the public, the project ensures that its innovations are accessible, understandable, and aligned with societal needs.
Conclusions
The NanoImmunoRT project is a pioneering effort to combine nanotechnology, radiotherapy, and immunotherapy into a single, synergistic treatment for GBM. By addressing hypoxia, radioresistance, and immune suppression, the project offers a holistic solution to the complex challenges of GBM therapy. Its results are expected to advance scientific knowledge, drive economic growth, and, most importantly, improve the lives of patients facing this aggressive disease. The project’s alignment with European health priorities and its commitment to sustainability and public engagement further underscore its potential for lasting impact.
Project objectives
The NanoImmunoRT project aims to develop a synergistic therapeutic strategy combining radiosensitization and TAM-targeted immunotherapy using multimodal therapeutic nanoparticles (MTNPs). The project leverages zeolite-based nanoparticles (LTL types) functionalized with manganese (Mn) and gadolinium (Gd) to enhance catalytic activity for oxygen generation and radiosensitization, and toll-like receptor (TLR) agonists (e.g. R848) to reprogram TAMs and stimulate anti-tumor immune responses.
The overall goal is to create a single nanoparticle platform that can:
- Decompose tumoral hydrogen peroxide (H2O2) into oxygen (O2), alleviating hypoxia and improving RT efficacy.
- Act as a radiosensitizer, enhancing DNA damage in cancer cells during RT.
- Re-educate TAMs from a pro-tumor (M2) to an anti-tumor (M1) phenotype, boosting the immune response against the tumor.
Project Pathway to Impact
The project introduces a novel MTNP formulation (LTL-Mn/Gd nanozeolites) with superior catalytic activity and radiosensitizing properties, addressing key limitations of current GBM therapies. In vitro and in vivo studies demonstrated that LTL-Mn/Gd nanozeolites enhance RT efficacy, reduce tumor hypoxia, and synergize with immunotherapy (R848), leading to significant tumor growth inhibition and prolonged survival in murine models. The project provides a proof-of-concept for combining RT and immunotherapy using MTNPs, paving the way for future preclinical studies in GBM and other hypoxic, immune-suppressive tumors.
Scale and significance of expected impacts
The project’s findings could help reshape the field of radioimmunotherapy, demonstrating the feasibility of using MTNPs to overcome hypoxia and immune suppression in GBM. The development of patentable MTNPs could stimulate innovation in the biotech sector, creating new markets and job opportunities. Moreover, by engaging with patients, clinicians, and the public, the project ensures that its innovations are accessible, understandable, and aligned with societal needs.
Conclusions
The NanoImmunoRT project is a pioneering effort to combine nanotechnology, radiotherapy, and immunotherapy into a single, synergistic treatment for GBM. By addressing hypoxia, radioresistance, and immune suppression, the project offers a holistic solution to the complex challenges of GBM therapy. Its results are expected to advance scientific knowledge, drive economic growth, and, most importantly, improve the lives of patients facing this aggressive disease. The project’s alignment with European health priorities and its commitment to sustainability and public engagement further underscore its potential for lasting impact.
Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far
The NanoImmunoRT project focused on developing multimodal therapeutic nanoparticles (MTNPs) to address the challenges of treating glioblastoma (GBM), particularly the limitations posed by hypoxia, radioresistance, and the immunosuppressive tumor microenvironment. The project's primary objective was to design and synthesize zeolite-based nanoparticles (LTL and RHO types) functionalized with manganese (Mn) and gadolinium (Gd) to enhance catalytic activity for oxygen generation and radiosensitization, as well as with Toll-like receptor (TLR) agonists (e.g. R848) to reprogram tumor-associated macrophages (TAMs).
In Work Package 1 (WP1), the project successfully synthesized and characterized LTL and RHO nanozeolites. At the partner lab LCS, Mn and Gd was incorporated using both ion exchange and a novel one-pot alanine/valine-assisted synthesis, which proved superior for Mn incorporation, resulting in nanoparticles with enhanced catalytic activity for oxygen generation from hydrogen peroxide (H2O2) decomposition. The successful incorporation of Mn and Gd was confirmed through SEM, XRD, and ICP-MS analyses. Additionally, a chemical grafting strategy was developed to attach resiquimod (R848) to the zeolite surface, enhancing the potential for TAM re-education. These achievements demonstrated the feasibility of creating nanoparticles with both radiosensitizing and immunomodulatory properties.
Work Package 2 (WP2) focused on evaluating the biocompatibility, immunotoxicity, and efficacy of the MTNPs using in vitro models. At the host lab ISTCT, cell viability was assessed in GL261 and RAW264.7 cells treated with various formulations of RHO and LTL nanozeolites. LTL nanozeolites exhibited lower toxicity compared to RHO nanozeolites, leading to their selection for subsequent experiments. LTL nanozeolites demonstrated dose- and time-dependent uptake in various cell types. The nanoparticles also showed effective penetration and accumulation in tumor spheroids, confirming their potential for targeted drug delivery. Furthermore, the combination of TLR agonists and radiotherapy was evaluated, revealing enhanced macrophage activation and production of pro-inflammatory markers. The radiosensitizing properties of LTL-Mn/Gd nanozeolites were confirmed under both normoxic and hypoxic conditions, highlighting their potential to overcome hypoxia-associated radioresistance.
In Work Package 3 (WP3), at the host lab ISTCT, the project transitioned to in vivo evaluations using preclinical mouse tumor models of glioma. An initial tolerability study identified 12.5 mg/kg as a safe dose for intravenous injections of LTL nanozeolites. Biodistribution studies in GL261 tumor-bearing mice demonstrated effective tumor targeting and prolonged retention of LTL-DiD nanozeolites. The therapeutic efficacy of LTL-Mn/Gd nanozeolites combined with radiotherapy was assessed, showing significant tumor growth inhibition and a 50% cure rate. Additionally, the combination of pIC+R848 immunotherapy with radiotherapy resulted in reduced tumor volumes and prolonged survival in both subcutaneous and orthotopic models. Preliminary imaging studies using ¹⁸F-MISO PET suggested that LTL-Mn/Gd nanozeolites effectively alleviate tumor hypoxia, further supporting their potential to enhance radiotherapy efficacy.
The outcomes of the NanoImmunoRT project are promising. The development of novel MTNP formulations with superior H2O2 catalytic activity and radiosensitizing properties could represent a significant advancement in cancer nanomedicine. The project provided preclinical evidence for the efficacy of combining radiotherapy and immunotherapy using MTNPs, demonstrating their potential to improve treatment outcomes for GBM. The results also showed that LTL-Mn/Gd nanozeolites can alleviate tumor hypoxia, which is a critical factor in radioresistance. Overall, the project achieved significant tumor growth inhibition and prolonged survival in preclinical models, paving the way for future translational studies.
In Work Package 1 (WP1), the project successfully synthesized and characterized LTL and RHO nanozeolites. At the partner lab LCS, Mn and Gd was incorporated using both ion exchange and a novel one-pot alanine/valine-assisted synthesis, which proved superior for Mn incorporation, resulting in nanoparticles with enhanced catalytic activity for oxygen generation from hydrogen peroxide (H2O2) decomposition. The successful incorporation of Mn and Gd was confirmed through SEM, XRD, and ICP-MS analyses. Additionally, a chemical grafting strategy was developed to attach resiquimod (R848) to the zeolite surface, enhancing the potential for TAM re-education. These achievements demonstrated the feasibility of creating nanoparticles with both radiosensitizing and immunomodulatory properties.
Work Package 2 (WP2) focused on evaluating the biocompatibility, immunotoxicity, and efficacy of the MTNPs using in vitro models. At the host lab ISTCT, cell viability was assessed in GL261 and RAW264.7 cells treated with various formulations of RHO and LTL nanozeolites. LTL nanozeolites exhibited lower toxicity compared to RHO nanozeolites, leading to their selection for subsequent experiments. LTL nanozeolites demonstrated dose- and time-dependent uptake in various cell types. The nanoparticles also showed effective penetration and accumulation in tumor spheroids, confirming their potential for targeted drug delivery. Furthermore, the combination of TLR agonists and radiotherapy was evaluated, revealing enhanced macrophage activation and production of pro-inflammatory markers. The radiosensitizing properties of LTL-Mn/Gd nanozeolites were confirmed under both normoxic and hypoxic conditions, highlighting their potential to overcome hypoxia-associated radioresistance.
In Work Package 3 (WP3), at the host lab ISTCT, the project transitioned to in vivo evaluations using preclinical mouse tumor models of glioma. An initial tolerability study identified 12.5 mg/kg as a safe dose for intravenous injections of LTL nanozeolites. Biodistribution studies in GL261 tumor-bearing mice demonstrated effective tumor targeting and prolonged retention of LTL-DiD nanozeolites. The therapeutic efficacy of LTL-Mn/Gd nanozeolites combined with radiotherapy was assessed, showing significant tumor growth inhibition and a 50% cure rate. Additionally, the combination of pIC+R848 immunotherapy with radiotherapy resulted in reduced tumor volumes and prolonged survival in both subcutaneous and orthotopic models. Preliminary imaging studies using ¹⁸F-MISO PET suggested that LTL-Mn/Gd nanozeolites effectively alleviate tumor hypoxia, further supporting their potential to enhance radiotherapy efficacy.
The outcomes of the NanoImmunoRT project are promising. The development of novel MTNP formulations with superior H2O2 catalytic activity and radiosensitizing properties could represent a significant advancement in cancer nanomedicine. The project provided preclinical evidence for the efficacy of combining radiotherapy and immunotherapy using MTNPs, demonstrating their potential to improve treatment outcomes for GBM. The results also showed that LTL-Mn/Gd nanozeolites can alleviate tumor hypoxia, which is a critical factor in radioresistance. Overall, the project achieved significant tumor growth inhibition and prolonged survival in preclinical models, paving the way for future translational studies.
Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)
The NanoImmunoRT project has made promising advances in treating glioblastoma (GBM) by developing nanozeolite-based multimodal therapeutic nanoparticles (MTNPs) that combine radiotherapy, immunotherapy, and nanotechnology. These nanoparticles, based on LTL zeolites and functionalized with manganese (Mn), gadolinium (Gd), and TLR agonists (e.g. R848), are designed to combat hypoxia, radioresistance, and immune suppression, key challenges in GBM treatment.
The project demonstrated that Mn-Val-LTL formulations exhibit high catalytic activity, effectively generating oxygen in hypoxic tumors by decomposing hydrogen peroxide (H2O2). In in vitro studies, these nanoparticles showed low toxicity, effective uptake in glioma and macrophage cells, and strong immunomodulatory effects when combined with radiotherapy. In vivo experiments confirmed their therapeutic potential, with LTL-Mn/Gd nanozeolites plus radiotherapy achieving a 50% cure rate in a mouse tumor model. These results highlight the potential of MTNPs to transform GBM treatment by addressing its most persistent challenges.
Broader Impacts and Future Potential
The NanoImmunoRT project holds innovative potential for the field of cancer nanomedicine. By demonstrating the feasibility of combining radiotherapy, immunotherapy, and nanotechnology, the project provides a proof-of-concept for a multimodal approach to GBM treatment. This could help redefine therapeutic strategies not only for GBM but also for other hypoxic and immune-suppressive tumors, offering new hope to patients who currently have limited treatment options.
From an economic and industrial perspective, the development of these novel MTNP formulations presents opportunities for commercialization. The scalable synthesis methods and biocompatibility of the nanozeolites make them attractive candidates for pharmaceutical development. Discussions with Zeonyx, a recently launched startup, and CNRS Innovation highlight the potential for patenting and commercializing this technology, which could lead to new therapeutic products and economic growth in the biotech sector.
Key Needs for Further Success
While the NanoImmunoRT project has made remarkable progress, several key steps are necessary to ensure its continued success and translation:
Further Research and Optimization: The next phase of research should focus on refining the MTNP formulations to enhance their therapeutic efficacy, stability, and scalability. Additional preclinical studies, including experiments with patient-derived organoids and xenograft models, will be essential to validate the nanoparticles' performance in more complex and clinically relevant settings.
Commercialization and Intellectual Property Protection: Protecting the intellectual property (IP) associated with the MTNP technology will be vital for attracting investment and facilitating commercialization. Collaborating with technology transfer offices, such as CNRS Innovation, will help navigate the patenting process and develop a robust business model for scalable production and distribution.
Standardization and Quality Assurance: Developing standardized protocols for the manufacturing, characterization, and testing of MTNPs will be essential for ensuring consistency, safety, and efficacy. These protocols will also facilitate regulatory approval and clinical adoption, making it easier to translate research findings into real-world treatments.
Public and Stakeholder Engagement: Continued public outreach and stakeholder engagement will be important for raising awareness about the project’s innovations and garnering support from patient advocacy groups, clinicians, and policymakers. Involving these stakeholders will help ensure that the development process remains patient-centered and ethically sound.
Conclusion
The NanoImmunoRT project has contributed to explore the potential of multimodal therapeutic nanoparticles to support radiotherapy, modulate immune responses, and address tumor hypoxia. While the findings are promising, they suggest new possibilities for treatments and encouraging directions for further research and innovation in the biotech field. The project’s outcomes highlight areas where continued investigation could lead to meaningful progress in tackling this challenging disease.
The project demonstrated that Mn-Val-LTL formulations exhibit high catalytic activity, effectively generating oxygen in hypoxic tumors by decomposing hydrogen peroxide (H2O2). In in vitro studies, these nanoparticles showed low toxicity, effective uptake in glioma and macrophage cells, and strong immunomodulatory effects when combined with radiotherapy. In vivo experiments confirmed their therapeutic potential, with LTL-Mn/Gd nanozeolites plus radiotherapy achieving a 50% cure rate in a mouse tumor model. These results highlight the potential of MTNPs to transform GBM treatment by addressing its most persistent challenges.
Broader Impacts and Future Potential
The NanoImmunoRT project holds innovative potential for the field of cancer nanomedicine. By demonstrating the feasibility of combining radiotherapy, immunotherapy, and nanotechnology, the project provides a proof-of-concept for a multimodal approach to GBM treatment. This could help redefine therapeutic strategies not only for GBM but also for other hypoxic and immune-suppressive tumors, offering new hope to patients who currently have limited treatment options.
From an economic and industrial perspective, the development of these novel MTNP formulations presents opportunities for commercialization. The scalable synthesis methods and biocompatibility of the nanozeolites make them attractive candidates for pharmaceutical development. Discussions with Zeonyx, a recently launched startup, and CNRS Innovation highlight the potential for patenting and commercializing this technology, which could lead to new therapeutic products and economic growth in the biotech sector.
Key Needs for Further Success
While the NanoImmunoRT project has made remarkable progress, several key steps are necessary to ensure its continued success and translation:
Further Research and Optimization: The next phase of research should focus on refining the MTNP formulations to enhance their therapeutic efficacy, stability, and scalability. Additional preclinical studies, including experiments with patient-derived organoids and xenograft models, will be essential to validate the nanoparticles' performance in more complex and clinically relevant settings.
Commercialization and Intellectual Property Protection: Protecting the intellectual property (IP) associated with the MTNP technology will be vital for attracting investment and facilitating commercialization. Collaborating with technology transfer offices, such as CNRS Innovation, will help navigate the patenting process and develop a robust business model for scalable production and distribution.
Standardization and Quality Assurance: Developing standardized protocols for the manufacturing, characterization, and testing of MTNPs will be essential for ensuring consistency, safety, and efficacy. These protocols will also facilitate regulatory approval and clinical adoption, making it easier to translate research findings into real-world treatments.
Public and Stakeholder Engagement: Continued public outreach and stakeholder engagement will be important for raising awareness about the project’s innovations and garnering support from patient advocacy groups, clinicians, and policymakers. Involving these stakeholders will help ensure that the development process remains patient-centered and ethically sound.
Conclusion
The NanoImmunoRT project has contributed to explore the potential of multimodal therapeutic nanoparticles to support radiotherapy, modulate immune responses, and address tumor hypoxia. While the findings are promising, they suggest new possibilities for treatments and encouraging directions for further research and innovation in the biotech field. The project’s outcomes highlight areas where continued investigation could lead to meaningful progress in tackling this challenging disease.