Abstract
Background
Although immunotherapy with immune checkpoint inhibitors (ICIs) has become a standard therapeutic strategy in colorectal cancer (CRC) exhibiting microsatellite instability-high, limited patients benefit from this new approach. To increase the efficacy of ICIs in CRC patients, it is crucial to control the function of immunosuppressive cells in the tumor microenvironment. M2-tumor-associated macrophages (TAMs) are key immunosuppressive cells and promote tumor growth, angiogenesis, and epithelial-mesenchymal transition. In the present study, we focused on the VEGF signaling pathway in M2-TAMs to control their inhibitory function.
Methods
We evaluated the population of M2-TAMs, the VEGF receptor 2 (VEGFR2) expression on M2-TAMs, and the correlation between HIF-1α-positive cells and VEGFR2 expression levels on M2-TAMs in CRC using the analysis of The Cancer Genome Atlas colorectal adenocarcinoma dataset (n = 592), the flow cytometry of freshly resected surgical specimens of CRC (n = 20), and the immunofluorescence staining of formalin-fixed paraffin-embedded whole tissue samples of CRC (n = 20). Furthermore, we performed a functional assay of M2 macrophages through the VEGF/VEGFR2 signaling pathway in vitro.
Results
The population of M2-TAMs and their VEGFR2 expression significantly increased in the tumor compared to the normal mucosa in the CRC patients. HIF1-α-positive cells significantly correlated with the VEGFR2 expression level of M2-TAMs. M2 macrophages induced by cytokines in vitro produced TGF-β1 through the VEGF/VEGFR2 signaling pathway.
Conclusions
Our results suggest that anti-VEGFR2 therapy may have therapeutic potential to control the immune inhibitory functions of M2-TAMs in CRC, resulting in enhanced efficacy of immunotherapy with ICIs.




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Abbreviations
- CRC:
-
Colorectal cancer
- EMT:
-
Epithelial-mesenchymal transition
- FFPE:
-
Formalin-fixed paraffin-embedded
- HIF:
-
Hypoxia-inducible factor
- HPD:
-
Hyper progressive disease
- ICIs:
-
Immune checkpoint inhibitors
- KDR:
-
Kinase insert domain receptor
- TAMs:
-
Tumor-associated macrophages
- TCGA:
-
The Cancer Genome Atlas
- VEGFR2:
-
VEGF receptor 2
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KM and KK contributed to the study conception and design. HO, WS, SF, HE, MS, ZS, TM, and SO contributed to the acquisition of patient samples. AKTM and SN performed flow cytometry and in vitro assay. AKTM, SN, and KS performed immunofluorescence staining. AKTM, KM, SN, HO, KS, WS, SF, HE, MS, ZS, TM, SO, and KK performed analysis and interpretation of results. AKTM, KM, and KK drafted the manuscript. All the authors are aware of and agree to the contents of the paper, as well as them being listed as authors on the paper.
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This study was conducted in accordance with the ethical principles of the 1964 Declaration of Helsinki and its later amendments and was approved by the Fukushima Medical University Research Ethics Committee (Reference Nos. 2289 and 29316).
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Written informed consent was obtained from all patients included in the study for the use of their specimens and clinical data for research and publication prior to collecting the specimens at Fukushima Medical University Hospital.
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Min, A.K.T., Mimura, K., Nakajima, S. et al. Therapeutic potential of anti-VEGF receptor 2 therapy targeting for M2-tumor-associated macrophages in colorectal cancer. Cancer Immunol Immunother 70, 289–298 (2021). https://doi.org/10.1007/s00262-020-02676-8
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DOI: https://doi.org/10.1007/s00262-020-02676-8