After 2030, more powerful anticancer drugs will be available [Science Now]

Postgraduate researcher Jiwoo Choi (first author) is using a pipette tool to transfer blood components into several wells (small containers) to culture them into macrophages [Photo = KRIST]



A domestic research team has found a clue to a new anticancer technology that can complement the existing CAR-T anticancer drug.



Dr. Jihoon Park's team at the Korea Research Institute of Chemical Technology (President Youngguk Lee) successfully produced CAR-M (Car-M, Car-Macrophage) by stably inserting anticancer genes into macrophages derived from human peripheral blood using lentivirus. It is



expected to be applied to the treatment of solid cancers in addition to blood cancers.



'Chimeric antigen receptor (CAR) T cell therapy' is a technology that extracts T cells, which are immune cells, from the body, genetically modifies them to attack specific cancer cells, and then injects them into the patient. It is very effective in treating some blood cancers such as leukemia, but has limitations in treating solid cancers such as lung cancer. Macrophages



are also a type of immune cell. T cells have difficulty penetrating solid tumors, but macrophages can easily penetrate them. Research is actively underway to apply macrophages as anticancer agents instead of T cells. However, existing macrophage utilization technologies have limitations in that anticancer gene modification occurs only for a short period of time, resulting in low therapeutic effects.



The research team developed various technologies that effectively deliver anticancer genes without damaging macrophages by using 'lentivirus' as a gene delivery method.



Generally, when a gene is implanted in a lentivirus and delivered to another cell, a cationic polymer 'polybrene' that thins the cell membrane is injected and mixed vigorously. This increases the cell penetration and gene transfer of lentivirus.



The problem is that when macrophages come into contact with polybrene, serious toxicity occurs, and the structure is damaged or the survival rate decreases during the vigorous rotation process.



The research team at the Chemical Research Institute has discovered a method to produce CAR-M anticancer agents. [Photo = Chemical Research Institute]



Instead of injecting polybrene or vigorous rotation, the research team increased the contact between lentivirus and macrophages from the original 1 hour and 30 minutes to 16 hours. As a result, the transmission of lentivirus occurred better than before without damaging macrophages.



In addition, it was confirmed that the gene transfer efficiency changed depending on the differentiation state of macrophages. Considering this, the gene transfer rate for cancer cell tracking was increased by waiting for 7 days while monocytes obtained from peripheral blood differentiated into macrophages.



The codon of the 'VSV-G protein', which acts as a key on the surface when lentivirus enters a cell, was optimized to further increase the gene transfer ability. A master key that can open various doors than the existing key was created, and the VSV-G production command was changed so that it could grow a lot on the surface.



Finally, the DNA sequence 'EF1a' was found and applied so that the anticancer gene delivered in the lentivirus could be well expressed in macrophages. As a result, the production of 'CAR macrophages' with anticancer function was maintained stably for up to 20 days after gene transfer without damaging the macrophages.



CAR macrophages were shown to effectively eliminate specific cancer cells. After expressing red fluorescent protein in Nalm6 and Raji cancer cells, which are representative cell lines of acute lymphoblastic leukemia and B-cell lymphoma, they checked it under an electron microscope after 5 days.



The red area was noticeably reduced. CAR macrophages engulfed and destroyed most of the cancer cells. The research team plans to develop mass production of CAR macrophages and high-efficiency treatment application technology through follow-up research.



The research team related to the chemical research institute explained, "It is significant in that it is the first case of improving the problem of low anticancer gene expression in macrophages obtained from peripheral blood using lentivirus."



Director Lee Young-guk of the chemical research institute said, "It will contribute to the diversification of immunotherapy by supplementing existing CAR T cell treatment methods."



This research was proven at the laboratory level, and in order to be commercialized and used as a treatment for patients, mass proliferation technology, etc. must be developed. The research team related to the chemical research institute predicted that it will be commercialized after 2030 as it is in the initial stage of laboratory-scale research and development.



This paper (title: Lentivirus-based production of human chimeric antigen receptor macrophages from peripheral blood) was published in the international academic journal 'Biomarker Research' in January 2025. Dr. Ji-Hoon Park of the Chemical Research Institute participated as the corresponding author, and Ji-Woo Choi, a postgraduate researcher, participated as the first author.





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