CAR-ENGINEERED HUMAN HEMATOPOIETIC STEM CELL MACROPHAGES AS AN INNOVATIVE PLATFORM FOR TUMOR CONTROL

@article{ovider2026carengineered,
  title = {{CAR-ENGINEERED HUMAN HEMATOPOIETIC STEM CELL MACROPHAGES AS AN INNOVATIVE PLATFORM FOR TUMOR CONTROL}},
  author = {Ian OVIDER and Livia FURQUIM and Carla FOGOLIN and Leonel JUNIOR and Viviane DA SILVA and Akash MATHEW and Elisa MATSUMURA and Lucas SANTOS and Vinicius PEREIRA and Gabriela EVANGELISTA and Theo DE OLIVEIRA and Samuel CAMPANELLI and Rossana FRANCISCO and Adriana PAES LEME and Tarcio BRAGA and Rafael ALMEIDA and Guilherme NADER and Vanderson ROCHA and Rodrigo Nalio RAMOS},
  journal = {Hematology, Transfusion and Cell Therapy},
  year = {2026},
  month = oct,
  doi = {10.1016/j.htct.2026.106833},
  url = {https://doi.org/10.1016/j.htct.2026.106833},
  note = {CC BY},
}

TY  - JOUR
TI  - CAR-ENGINEERED HUMAN HEMATOPOIETIC STEM CELL MACROPHAGES AS AN INNOVATIVE PLATFORM FOR TUMOR CONTROL
AU  - Ian OVIDER
AU  - Livia FURQUIM
AU  - Carla FOGOLIN
AU  - Leonel JUNIOR
AU  - Viviane DA SILVA
AU  - Akash MATHEW
AU  - Elisa MATSUMURA
AU  - Lucas SANTOS
AU  - Vinicius PEREIRA
AU  - Gabriela EVANGELISTA
AU  - Theo DE OLIVEIRA
AU  - Samuel CAMPANELLI
AU  - Rossana FRANCISCO
AU  - Adriana PAES LEME
AU  - Tarcio BRAGA
AU  - Rafael ALMEIDA
AU  - Guilherme NADER
AU  - Vanderson ROCHA
AU  - Rodrigo Nalio RAMOS
PY  - 2026
DA  - 2026/10/23
JO  - Hematology, Transfusion and Cell Therapy
DO  - 10.1016/j.htct.2026.106833
UR  - https://doi.org/10.1016/j.htct.2026.106833
AB  - Introduction: CAR-T cell therapy has emerged as a highly promising strategy for the treatment of hematological malignancies. However, its efficacy against solid tumors remains limited, mainly due to the establishment of an immunosuppressive tumor microenvironment that impairs CAR-T cell trafficking and functional activation. Given their intrinsic ability to infiltrate solid tumors and interact with several components of the tumor microenvironment, macrophages have emerged as promising candidates for CAR-based cell therapy. However, sources for obtaining macrophages are limited due to their low proliferative capacity, and genetically reprogramming mature monocytes/ macrophages is extremely challenging. Objective: We aimed to generate functional macrophages from human umbilical cord blood-derived CD34+ hematopoietic stem cells (HSCs) engineered to express CARs. Methods: Fresh isolated HSC were in vitro expanded and transduced with a second-generation chimeric lentiviral vector for anti-HER2. Subsequently, expanded CAR-HSC were differentiated into macrophages (MacCD34) in the presence of GM-CSF, M-CSF, and IL-3 for 10 days. Results: Approximately 50% of the CAR-MacCD34 population expressed anti-HER2 CARs and displayed nearly 95% of viability throughout differentiation. Mass spectrometry (MS) and multiparametric flow cytometry analysis revealed upregulation of proteins associated with phagocytosis, matrix remodeling, and degradation, indicating enhanced tumor infiltration potential. In vitro, CAR-MacCD34 showed a significantly improved ability to eliminate and phagocytose HER2+ tumor cells when compared to untransduced MacCD34 cells. Additionally, using high definition microscopy we noted that CAR-MacCD34 cells that phagocytosed cancer cells showed increased nuclear translocation of NF-KB, suggesting CAR-mediated intracellular signaling. To assess functionality in a more physiologically relevant context, we used tumor spheroids embedded in a dense 3D collagen matrix. Confocal microscopy and live-cell imaging revealed that CAR-MacCD34 cells efficiently migrated toward, and infiltrated tumor spheroids compared to untransduced MacCD34 cells. In addition, multiple tumor cell phagocytosis events mediated by CAR-MacCD34 cells were observed within the 3D model. We further evaluated the anti-tumor activity of CAR-MacCD34 cells in zebrafish larvae models of HER2+ tumors. Our results showed that CAR-MacCD34 cells persisted for over 8 days post-injection and promoted a significant reduction in tumor burden compared to untransduced MacCD34 cells. Conclusions: Collectively, our findings support a novel CAR-macrophage therapeutic approach with promising clinical potential, based on a renewable and accessible cellular source. Optimizing CAR-MacCD34 functionality in combination with existing therapies may contribute to durable anti-tumor responses and improved control of tumors. This work was supported by FAPESP, CNPq and PRONON.
SN  - 2531-1379
SN  - 2531-1387
ER  -