Periodic Reporting for period 1 - MONONANOCHEM (In vivo engineering of monocytes loaded with nano-chemotherapeutic formulations: a novel live-cell mediated drug delivery system for the treatment of cancer)
Okres sprawozdawczy: 2017-01-01 do 2018-12-31
The success of current anti-tumoral therapies largely depends on the availability of appropriate drug delivery strategies, which have to allow the penetration of these drugs through the tumoral microenvironment. The knowledge related to the cellular heterogeneity of the tumors has advanced enormously in the last years. The recent studies showed that tumor associated macrophages (TAMs) can represent up to 50% of the tumor mass. TAMs are derived from circulating monocytes, which are recruited from the peripheral blood towards the tumor in response to a broad range of molecular signals. Recently several studies are focused on the development of therapeutic strategies targeting TAMs, however, the possibility to use monocytes, with excellent ability to infiltrate the abnormal tumoral tissue, to deliver chemotherapeutic drugs into the tumor has been hardly explored.
Here, we aim to use the host monocytes, with high ability to infiltrate the tumoral tissues, to deliver chemotherapeutic nanoformulations into the bulk of the tumor. We have developed Nano-Formulations consisting in Poly(Lactide-co-Glycolide) (PLGA) nanostructures, allowing the loading of chemotherapeutic drugs into monocytes. Cytotoxic drugs inducing immunogenic cell death (Doxorubicin), and thus the recruitment of new monocytes, were used to enhance the efficacy of the therapy. Overall, we used Monocytes loaded with ChemoTherapeutic Nano-Formulations (MCTNFs) as a new strategy to deliver cytotoxic drugs towards the tumoral cells, taking advantage of the infiltration ability of the monocytes into tumor microenvironment. The MCTNFs were tested using appropriate in vitro 2D tumor models and in vivo murine tumor models. We expect that this new live-cell mediated drug delivery system will provide greater progress in the treatment of cancer.
Here, we aim to use the host monocytes, with high ability to infiltrate the tumoral tissues, to deliver chemotherapeutic nanoformulations into the bulk of the tumor. We have developed Nano-Formulations consisting in Poly(Lactide-co-Glycolide) (PLGA) nanostructures, allowing the loading of chemotherapeutic drugs into monocytes. Cytotoxic drugs inducing immunogenic cell death (Doxorubicin), and thus the recruitment of new monocytes, were used to enhance the efficacy of the therapy. Overall, we used Monocytes loaded with ChemoTherapeutic Nano-Formulations (MCTNFs) as a new strategy to deliver cytotoxic drugs towards the tumoral cells, taking advantage of the infiltration ability of the monocytes into tumor microenvironment. The MCTNFs were tested using appropriate in vitro 2D tumor models and in vivo murine tumor models. We expect that this new live-cell mediated drug delivery system will provide greater progress in the treatment of cancer.
We have demonstrated the proof-of-concept of the new anti-tumor drug delivery strategy using monocytes as a cargo. First, we have designed selected the Poly(Lactide-co-Glycolide) (PLGA) based the Nano-Formulations (NFs) to be loaded into monocytes, and established the protocol to produce fluorescent dye conjugated PLGA NFs. Then using mouse fibrosarcoma model, we have demonstrated the successful recruitment of human monocytes to the tumor site. This protocol was developed to confirm that the monocytes ingested NFs reach to the tumor site. Importantly, the distribution of the injected monocytes was different from that of injected free NFs, which mainly accumulate in liver and lung. This is one of the main achievements in our project, and proved that this strategy of using monocytes recruited in tumor tissues has the possibility to deliver anti-tumor chemotherapy.
As an anti-tumor drug, we selected Doxorubicin (Doxo), which is well used in oncology. We have demonstrated that the Doxo encapsulated PLGA NFs (PLGA-Doxo) have anti-tumor effect with in vitro experiments, as well as with Doxo encapsulated micelle based NFs (Doxo-micelle). We have examined the anti-tumor effect of Doxo-micelle using our mouse tumor model, and the result was significantly positive. Interestingly, the monocytes ingested Doxo-micelles exhibited strong anti-tumor activity. These micelles conceived on the amount of Doxo was much lower than the effective concentration of Doxo alone. This result indicated that our strategy achieves specific targeting of tumors and contributes a possible treatment to avoid unwanted drug toxicity.
As an anti-tumor drug, we selected Doxorubicin (Doxo), which is well used in oncology. We have demonstrated that the Doxo encapsulated PLGA NFs (PLGA-Doxo) have anti-tumor effect with in vitro experiments, as well as with Doxo encapsulated micelle based NFs (Doxo-micelle). We have examined the anti-tumor effect of Doxo-micelle using our mouse tumor model, and the result was significantly positive. Interestingly, the monocytes ingested Doxo-micelles exhibited strong anti-tumor activity. These micelles conceived on the amount of Doxo was much lower than the effective concentration of Doxo alone. This result indicated that our strategy achieves specific targeting of tumors and contributes a possible treatment to avoid unwanted drug toxicity.
We have demonstrated to link developments in the fields of cancer, immunology and nanotechnology. This project, MONONANOCHEM aimed to develop an innovative live cell-mediated drug delivery strategy for the treatment of cancer, using nanotechnology as a tool for the in vivo development of the therapy (nano-chemotherapeutic loaded monocytes). For this purpose, in the frame of this project, we have managed to gather the support of leading researchers in the fields of cancer immunotherapy and nanomedicine. Our work connected and supported the transfer of different skills and knowledge within the host laboratory (Instituto Clinico Humanitas; ICH) and partner laboratories (Universidade de Santiago de Compostela; USC, Biccoca University; BU). This will be beneficial for the institutions, for the EU, enhancing the collaboration of excellent researchers from different countries within EU (Italy and Spain). We hope that these interactions will progress through related projects in the near future between the participant organizations, and joining of additional institutions and researchers at the EU level.
From the experience during the time of the project, the researcher has accepted benefits both scientifically and personally. We have published 2 original articles and 1 review in the international journal, and now we are preparing another 2 original articles. This will definitely contribute to the researcher’s carrier and increase chances to develop his scientific carrier. The skills and knowledge obtained from the time of the action will also make his professional capacity wider, which will help for the researcher to work in the field of cancer and immunology in the future. Though the management of the project for 2 years, the researcher has learned how to organize both short and long term experimental plan for the project, including the risk management of the project. In the future, this experience will help to lead bigger projects as a senior scientist.
Long-term objectives of this project provided new perspectives for long-term research in the fields of cancer immunotherapy and drug delivery within EU, with a special focus in linking both disciplines. Scientific contacts between the researcher and the project enhanced the incipient ties between these established fields. This project is in line with the Horizon2020 priorities, promoting: Excellence in Science (training of excellent researchers), Industrial Leadership (nanotechnologies) and Societal Challenges (health). The novel approaches for the design of therapies, methodologies, evaluation methods and research experience arising along this project has provided a reference for the use of innate immune cells as drug carriers. This will suppose a breakthrough in the treatment of cancer and possibly for the cure of other diseases. This action will contribute to improve EU excellence due to: first, the high level of training received by the researcher at ICH, and second, by the integration of the researcher within EU. Having the support from this project allowed the researcher a successful development of his career as an independent researcher within EU. Although the current project has ended, the ultimate goal of this project is, providing the benefits to the institution and all participants joined in this proposal, to provide solutions for the patients with cancer and improve their life.
From the experience during the time of the project, the researcher has accepted benefits both scientifically and personally. We have published 2 original articles and 1 review in the international journal, and now we are preparing another 2 original articles. This will definitely contribute to the researcher’s carrier and increase chances to develop his scientific carrier. The skills and knowledge obtained from the time of the action will also make his professional capacity wider, which will help for the researcher to work in the field of cancer and immunology in the future. Though the management of the project for 2 years, the researcher has learned how to organize both short and long term experimental plan for the project, including the risk management of the project. In the future, this experience will help to lead bigger projects as a senior scientist.
Long-term objectives of this project provided new perspectives for long-term research in the fields of cancer immunotherapy and drug delivery within EU, with a special focus in linking both disciplines. Scientific contacts between the researcher and the project enhanced the incipient ties between these established fields. This project is in line with the Horizon2020 priorities, promoting: Excellence in Science (training of excellent researchers), Industrial Leadership (nanotechnologies) and Societal Challenges (health). The novel approaches for the design of therapies, methodologies, evaluation methods and research experience arising along this project has provided a reference for the use of innate immune cells as drug carriers. This will suppose a breakthrough in the treatment of cancer and possibly for the cure of other diseases. This action will contribute to improve EU excellence due to: first, the high level of training received by the researcher at ICH, and second, by the integration of the researcher within EU. Having the support from this project allowed the researcher a successful development of his career as an independent researcher within EU. Although the current project has ended, the ultimate goal of this project is, providing the benefits to the institution and all participants joined in this proposal, to provide solutions for the patients with cancer and improve their life.