Periodic Reporting for period 4 - McHAP (Entrapment of Hypoxic Cancer by Macrophages Loaded with HAP)
Reporting period: 2021-07-01 to 2022-12-31
According to the statistics every second person will be diagnosed with cancer during their lifetime. One out of every four will die because of this disease. Despite enormous knowledge improvement, the prognosis in cases with advanced tumors is still unsatisfactory. Firstly, only 1-2% of the administered drug is delivered to the solid tumor, while the rest goes to other tissues causing side effects. These 1-2% reaches predominantly well vascularized regions and therefore, solid tumors initially often respond well to conventional anti-cancer therapy decreasing the tumor mass. However, subsequent regrowth or metastasis after radio- or chemotherapy remains a serious problem in clinical oncology due to difficulties in penetration of the anticancer drug to the most hypoxic areas where cancer cells may remain. Moreover, chemotherapy or radiotherapy may cause additional formation of large hypoxic areas, making the treatment of tumor regrowth even more difficult. The fact that the efficacy of anticancer therapy is limited by the presence of hypoxic tumor cells has led to the introduction of a variety of therapeutic approaches aimed at elimination of these hypoxic areas. Therefore, improving drug delivery to hypoxic regions might be a ground breaking strategy in oncology. A key consequence of hypoxia is an increase in infiltration of macrophages, which undergo polarization toward tumor associated macrophages (TAMs) with M2 phenotype and paradoxically actively facilitate tumor development and spread to distant sites. Because macrophages are continuously recruited into the tumor mass and actively migrate to avascular and hypoxic regions, they represent attractive vehicles to deliver genes or other particles into the tumor.
The general objective of the project is to investigate novel macrophages-based delivery system of anticancer drugs encapsulated in ferritin cages into the hypoxic tumor areas and to investigate the underlying molecular mechanisms. Macrophages are physiologically attracted by the tumor, then they have ability to migrate from blood vessels and infiltrate avascular and hypoxic areas. Therefore they might constitute a unique delivery system of drug containing particles to the hypoxic parts of the tumor. Caged architecture of ferritin allows for efficient drug encapsulation. Macrophages are able to specifically and actively transfer ferritins (loaded with component of choice) to cancer cells. That macrophages loaded with ferritin containing anticancer drug could be an effective tool to smuggle drugs inside the tumor mass. The final effect will be the novel and effective macrophage-based delivery system of ferritin encapsulated drug that will constitute a starting point to development of a novel treatment strategy.
The general objective of the project is to investigate novel macrophages-based delivery system of anticancer drugs encapsulated in ferritin cages into the hypoxic tumor areas and to investigate the underlying molecular mechanisms. Macrophages are physiologically attracted by the tumor, then they have ability to migrate from blood vessels and infiltrate avascular and hypoxic areas. Therefore they might constitute a unique delivery system of drug containing particles to the hypoxic parts of the tumor. Caged architecture of ferritin allows for efficient drug encapsulation. Macrophages are able to specifically and actively transfer ferritins (loaded with component of choice) to cancer cells. That macrophages loaded with ferritin containing anticancer drug could be an effective tool to smuggle drugs inside the tumor mass. The final effect will be the novel and effective macrophage-based delivery system of ferritin encapsulated drug that will constitute a starting point to development of a novel treatment strategy.
The high level aim of the project was to identify molecular basis of the newly discovered phenomenon and investigate if it can be used in practice as a novel anti-cancer therapy.
The more specific aims of the project were as follow:
1. Explore full mechanisms and reason of Ft uptake by macrophages in vitro and Ft fate in these cells in standard conditions and hypoxia.
Within this Task we identified the receptor responsible for ferritin uptake by macrophages. We confimred both using molecular biology tools and biochemical tools that the specific recpetor binds ferritin and in a clathrin-dependent manner it is internalized inside the macrophage. In macrophages ferritin goes to the early endosomes and later on to the lysosomes. There is no difference in ferritin uptake in different oxygen conditions.
These results are included in the PhD thesis of Ms. Małgorzata Kubiak (PhD student within the McHAP project) and will be submitted to publication in Q1 2023.
2. Characterization of the Ft transport from macrophages to cancer cells in standard conditions and oxidative stress.
Within this Task we identified the mechanisms of newly discovered biological phenomenon that ferritin is transfered from macrophages to cancer cells. We identified that a close interaction between both cells is required for transfer to occur and we have indentified the molecular players involved in this process. We showed that not only the exogenous ferritin is transfered, but also the intracellular protein. Moreover, we identified the fate of ferritin in cancer cell after the transfer.
These results are included in the PhD thesis of Mr. Maciej Białasek (PhD student within the McHAP project) and will be submitted to publication in Q1 2023.
3. Investigation of the safety of macrophages trafficking to the tumour and metastases.
Within this Task we showed high efficacy of macrophage migration toward tumors and its metastases. Moreover, we showed that macrophage administration - autologus or allogeneic is safe.
4. Investigation of the efficacy of the innovative macrophage-Ft therapy.
Within this Task we developed novel formulations of existing drugs based on encapsulation with Ft and showed their efficacy in reducing tumor burden.
These results are included in the PhD thesis of Ms. Małgorzata Górczak (PhD student within the McHAP project) and will be submitted to publication in Q1 2023.
All the Tasks have been completed. We have identified mechanisms by which macrophages take up ferritins and we identified the mechanism of ferritin transfer from macrophage to cancer cell. This macrophage therapy is safe and the transfer is sufficient enough to kill cancer cells by the drug loaded inside ferritin cavity. The overal idea can be used and further develop as anticancer therapy.
The more specific aims of the project were as follow:
1. Explore full mechanisms and reason of Ft uptake by macrophages in vitro and Ft fate in these cells in standard conditions and hypoxia.
Within this Task we identified the receptor responsible for ferritin uptake by macrophages. We confimred both using molecular biology tools and biochemical tools that the specific recpetor binds ferritin and in a clathrin-dependent manner it is internalized inside the macrophage. In macrophages ferritin goes to the early endosomes and later on to the lysosomes. There is no difference in ferritin uptake in different oxygen conditions.
These results are included in the PhD thesis of Ms. Małgorzata Kubiak (PhD student within the McHAP project) and will be submitted to publication in Q1 2023.
2. Characterization of the Ft transport from macrophages to cancer cells in standard conditions and oxidative stress.
Within this Task we identified the mechanisms of newly discovered biological phenomenon that ferritin is transfered from macrophages to cancer cells. We identified that a close interaction between both cells is required for transfer to occur and we have indentified the molecular players involved in this process. We showed that not only the exogenous ferritin is transfered, but also the intracellular protein. Moreover, we identified the fate of ferritin in cancer cell after the transfer.
These results are included in the PhD thesis of Mr. Maciej Białasek (PhD student within the McHAP project) and will be submitted to publication in Q1 2023.
3. Investigation of the safety of macrophages trafficking to the tumour and metastases.
Within this Task we showed high efficacy of macrophage migration toward tumors and its metastases. Moreover, we showed that macrophage administration - autologus or allogeneic is safe.
4. Investigation of the efficacy of the innovative macrophage-Ft therapy.
Within this Task we developed novel formulations of existing drugs based on encapsulation with Ft and showed their efficacy in reducing tumor burden.
These results are included in the PhD thesis of Ms. Małgorzata Górczak (PhD student within the McHAP project) and will be submitted to publication in Q1 2023.
All the Tasks have been completed. We have identified mechanisms by which macrophages take up ferritins and we identified the mechanism of ferritin transfer from macrophage to cancer cell. This macrophage therapy is safe and the transfer is sufficient enough to kill cancer cells by the drug loaded inside ferritin cavity. The overal idea can be used and further develop as anticancer therapy.
Results of this project is to identify molecular basis of the newly discovered phenomenon (TRAIN) that has not been described before. Therefore these results constitute novel and original input into the basic science in the field of cancer immunology and immunotherapy.
Results of this project are important for development of innovative cell-based drug delivery system to solid tumors and especially their hypoxic sites located away from blood vessels. This therapy may constitute a ground-breaking approach for targeting solid tumors.
Goals of the project are reached in accordance to the envisioned time-frames. Moreover, based on this project two ERC Proof of Concept grants have been obtained. Additionally, based on one of the ERC PoC grants, the EIC Transition project has been received for further development of the anticancer therapy.
Results of this project are important for development of innovative cell-based drug delivery system to solid tumors and especially their hypoxic sites located away from blood vessels. This therapy may constitute a ground-breaking approach for targeting solid tumors.
Goals of the project are reached in accordance to the envisioned time-frames. Moreover, based on this project two ERC Proof of Concept grants have been obtained. Additionally, based on one of the ERC PoC grants, the EIC Transition project has been received for further development of the anticancer therapy.