Skip to main content

Advertisement

SpringerLink
Log in

Therapeutic potential of anti-VEGF receptor 2 therapy targeting for M2-tumor-associated macrophages in colorectal cancer

  • Original Article
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

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

References

  1. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A (2018) Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 68(6):394–424. https://doi.org/10.3322/caac.21492

    Article  PubMed  Google Scholar 

  2. Cercek A, Roxburgh CSD, Strombom P, Smith JJ, Temple LKF, Nash GM, Guillem JG, Paty PB, Yaeger R, Stadler ZK, Seier K, Gonen M, Segal NH, Reidy DL, Varghese A, Shia J, Vakiani E, Wu AJ, Crane CH, Gollub MJ, Garcia-Aguilar J, Saltz LB, Weiser MR (2018) Adoption of total neoadjuvant therapy for locally advanced rectal cancer. JAMA Oncol. https://doi.org/10.1001/jamaoncol.2018.0071

    Article  PubMed  PubMed Central  Google Scholar 

  3. Gollins S, Sebag-Montefiore D (2016) Neoadjuvant treatment strategies for locally advanced rectal cancer. Clin Oncol 28(2):146–151. https://doi.org/10.1016/j.clon.2015.11.003

    Article  CAS  Google Scholar 

  4. Overman MJ, Lonardi S, Wong KYM, Lenz HJ, Gelsomino F, Aglietta M, Morse MA, Van Cutsem E, McDermott R, Hill A, Sawyer MB, Hendlisz A, Neyns B, Svrcek M, Moss RA, Ledeine JM, Cao ZA, Kamble S, Kopetz S, Andre T (2018) Durable clinical benefit with nivolumab plus ipilimumab in DNA mismatch repair-deficient/microsatellite instability-high metastatic colorectal cancer. J Clin Oncol 36(8):773–779. https://doi.org/10.1200/jco.2017.76.9901

    Article  CAS  PubMed  Google Scholar 

  5. Overman MJ, McDermott R, Leach JL, Lonardi S, Lenz HJ, Morse MA, Desai J, Hill A, Axelson M, Moss RA, Goldberg MV, Cao ZA, Ledeine JM, Maglinte GA, Kopetz S, Andre T (2017) Nivolumab in patients with metastatic DNA mismatch repair-deficient or microsatellite instability-high colorectal cancer (CheckMate 142): an open-label, multicentre, phase 2 study. Lancet Oncol 18(9):1182–1191. https://doi.org/10.1016/S1470-2045(17)30422-9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Shibuya KC, Goel VK, Xiong W, Sham JG, Pollack SM, Leahy AM, Whiting SH, Yeh MM, Yee C, Riddell SR, Pillarisetty VG (2014) Pancreatic ductal adenocarcinoma contains an effector and regulatory immune cell infiltrate that is altered by multimodal neoadjuvant treatment. PLoS ONE 9(5):e96565. https://doi.org/10.1371/journal.pone.0096565

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. De Monte L, Reni M, Tassi E, Clavenna D, Papa I, Recalde H, Braga M, Di Carlo V, Doglioni C, Protti MP (2011) Intratumor T helper type 2 cell infiltrate correlates with cancer-associated fibroblast thymic stromal lymphopoietin production and reduced survival in pancreatic cancer. J Exp Med 208(3):469–478. https://doi.org/10.1084/jem.20101876

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Qian BZ, Pollard JW (2010) Macrophage diversity enhances tumor progression and metastasis. Cell 141(1):39–51. https://doi.org/10.1016/j.cell.2010.03.014

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Condeelis J, Pollard JW (2006) Macrophages: obligate partners for tumor cell migration, invasion, and metastasis. Cell 124(2):263–266. https://doi.org/10.1016/j.cell.2006.01.007

    Article  CAS  PubMed  Google Scholar 

  10. Pollard JW (2004) Tumour-educated macrophages promote tumour progression and metastasis. Nat Rev Cancer 4(1):71–78. https://doi.org/10.1038/nrc1256

    Article  CAS  PubMed  Google Scholar 

  11. Yang L, Zhang Y (2017) Tumor-associated macrophages: from basic research to clinical application. J Hematol Oncol 10(1):58. https://doi.org/10.1186/s13045-017-0430-2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Nakajima S, Koh V, Kua LF, So J, Davide L, Lim KS, Petersen SH, Yong WP, Shabbir A, Kono K (2016) Accumulation of CD11c+CD163+ adipose tissue macrophages through upregulation of intracellular 11beta-HSD1 in human obesity. J Immunol 197(9):3735–3745. https://doi.org/10.4049/jimmunol.1600895

    Article  CAS  PubMed  Google Scholar 

  13. Hinshaw DC, Shevde LA (2019) The tumor microenvironment innately modulates cancer progression. Can Res 79(18):4557–4566. https://doi.org/10.1158/0008-5472.Can-18-3962

    Article  CAS  Google Scholar 

  14. Mantovani A, Sica A (2010) Macrophages, innate immunity and cancer: balance, tolerance, and diversity. Curr Opin Immunol 22(2):231–237. https://doi.org/10.1016/j.coi.2010.01.009

    Article  CAS  PubMed  Google Scholar 

  15. Biswas SK, Gangi L, Paul S, Schioppa T, Saccani A, Sironi M, Bottazzi B, Doni A, Vincenzo B, Pasqualini F, Vago L, Nebuloni M, Mantovani A, Sica A (2006) A distinct and unique transcriptional program expressed by tumor-associated macrophages (defective NF-kappaB and enhanced IRF-3/STAT1 activation). Blood 107(5):2112–2122. https://doi.org/10.1182/blood-2005-01-0428

    Article  CAS  PubMed  Google Scholar 

  16. Batlle E, Massague J (2019) Transforming growth factor-beta signaling in immunity and cancer. Immunity 50(4):924–940. https://doi.org/10.1016/j.immuni.2019.03.024

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Kikuchi T, Mimura K, Ashizawa M, Okayama H, Endo E, Saito K, Sakamoto W, Fujita S, Endo H, Saito M, Momma T, Saze Z, Ohki S, Shimada K, Yoshimura K, Tsunoda T, Kono K (2020) Characterization of tumor-infiltrating immune cells in relation to microbiota in colorectal cancers. Cancer Immunol Immunother 69(1):23–32. https://doi.org/10.1007/s00262-019-02433-6

    Article  CAS  PubMed  Google Scholar 

  18. Wheeler KC, Jena MK, Pradhan BS, Nayak N, Das S, Hsu CD, Wheeler DS, Chen K, Nayak NR (2018) VEGF may contribute to macrophage recruitment and M2 polarization in the decidua. PLoS ONE 13(1):e0191040. https://doi.org/10.1371/journal.pone.0191040

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Gao J, Aksoy BA, Dogrusoz U, Dresdner G, Gross B, Sumer SO, Sun Y, Jacobsen A, Sinha R, Larsson E, Cerami E, Sander C, Schultz N (2013) Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal. Sci Signal 6(269):pl1. https://doi.org/10.1126/scisignal.2004088

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Cerami E, Gao J, Dogrusoz U, Gross BE, Sumer SO, Aksoy BA, Jacobsen A, Byrne CJ, Heuer ML, Larsson E, Antipin Y, Reva B, Goldberg AP, Sander C, Schultz N (2012) The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. Cancer Discov 2(5):401–404. https://doi.org/10.1158/2159-8290.Cd-12-0095

    Article  PubMed  Google Scholar 

  21. Nakayama Y, Mimura K, Tamaki T, Shiraishi K, Kua LF, Koh V, Ohmori M, Kimura A, Inoue S, Okayama H, Suzuki Y, Nakazawa T, Ichikawa D, Kono K (2019) PhosphoSTAT1 expression as a potential biomarker for antiPD1/antiPDL1 immunotherapy for breast cancer. Int J Oncol 54(6):2030–2038. https://doi.org/10.3892/ijo.2019.4779

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Kikuchi T, Mimura K, Okayama H, Nakayama Y, Saito K, Yamada L, Endo E, Sakamoto W, Fujita S, Endo H, Saito M, Momma T, Saze Z, Ohki S, Kono K (2019) A subset of patients with MSS/MSI-low-colorectal cancer showed increased CD8(+) TILs together with up-regulated IFN-gamma. Oncol Lett 18(6):5977–5985. https://doi.org/10.3892/ol.2019.10953

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Nakayama Y, Mimura K, Kua LF, Okayama H, Min AKT, Saito K, Hanayama H, Watanabe Y, Saito M, Momma T, Saze Z, Ohki S, Suzuki Y, Ichikawa D, Yong WP, Kono K (2020) Immune suppression caused by PD-L2 expression on tumor cells in gastric cancer. Gastric Cancer. https://doi.org/10.1007/s10120-020-01079-z

    Article  PubMed  PubMed Central  Google Scholar 

  24. Zarif JC, Hernandez JR, Verdone JE, Campbell SP, Drake CG, Pienta KJ (2016) A phased strategy to differentiate human CD14+monocytes into classically and alternatively activated macrophages and dendritic cells. Biotechniques 61(1):33–41. https://doi.org/10.2144/000114435

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Onozawa H, Saito M, Saito K, Kanke Y, Watanabe Y, Hayase S, Sakamoto W, Ishigame T, Momma T, Ohki S, Takenoshita S (2017) Annexin A1 is involved in resistance to 5-FU in colon cancer cells. Oncol Rep 37(1):235–240. https://doi.org/10.3892/or.2016.5234

    Article  PubMed  Google Scholar 

  26. Okano M, Kumamoto K, Saito M, Onozawa H, Saito K, Abe N, Ohtake T, Takenoshita S (2015) Upregulated Annexin A1 promotes cellular invasion in triple-negative breast cancer. Oncol Rep 33(3):1064–1070. https://doi.org/10.3892/or.2015.3720

    Article  CAS  PubMed  Google Scholar 

  27. Thar Min AK, Okayama H, Saito M, Ashizawa M, Aoto K, Nakajima T, Saito K, Hayase S, Sakamoto W, Tada T, Hanayama H, Saze Z, Momma T, Ohki S, Sato Y, Motoyama S, Mimura K, Kono K (2018) Epithelial-mesenchymal transition-converted tumor cells can induce T-cell apoptosis through upregulation of programmed death ligand 1 expression in esophageal squamous cell carcinoma. Cancer Med. https://doi.org/10.1002/cam4.1564

    Article  PubMed  PubMed Central  Google Scholar 

  28. Ashizawa M, Okayama H, Ishigame T, Thar Min AK, Saito K, Ujiie D, Murakami Y, Kikuchi T, Nakayama Y, Noda M, Tada T, Endo H, Fujita S, Sakamoto W, Saito M, Saze Z, Momma T, Ohki S, Mimura K, Kono K (2019) miRNA-148a-3p regulates immunosuppression in DNA mismatch repair-deficient colorectal cancer by targeting PD-L1. Mol Cancer Res 17(6):1403–1413. https://doi.org/10.1158/1541-7786.Mcr-18-0831

    Article  CAS  PubMed  Google Scholar 

  29. Etzerodt A, Tsalkitzi K, Maniecki M, Damsky W, Delfini M, Baudoin E, Moulin M, Bosenberg M, Graversen JH, Auphan-Anezin N, Moestrup SK, Lawrence T (2019) Specific targeting of CD163(+) TAMs mobilizes inflammatory monocytes and promotes T cell-mediated tumor regression. J Exp Med 216(10):2394–2411. https://doi.org/10.1084/jem.20182124

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Komohara Y, Jinushi M, Takeya M (2014) Clinical significance of macrophage heterogeneity in human malignant tumors. Cancer Sci 105(1):1–8. https://doi.org/10.1111/cas.12314

    Article  CAS  PubMed  Google Scholar 

  31. Laoui D, Van Overmeire E, Di Conza G, Aldeni C, Keirsse J, Morias Y, Movahedi K, Houbracken I, Schouppe E, Elkrim Y, Karroum O, Jordan B, Carmeliet P, Gysemans C, De Baetselier P, Mazzone M, Van Ginderachter JA (2014) Tumor hypoxia does not drive differentiation of tumor-associated macrophages but rather fine-tunes the M2-like macrophage population. Can Res 74(1):24–30. https://doi.org/10.1158/0008-5472.Can-13-1196

    Article  CAS  Google Scholar 

  32. Colegio OR, Chu NQ, Szabo AL, Chu T, Rhebergen AM, Jairam V, Cyrus N, Brokowski CE, Eisenbarth SC, Phillips GM, Cline GW, Phillips AJ, Medzhitov R (2014) Functional polarization of tumour-associated macrophages by tumour-derived lactic acid. Nature 513(7519):559–563. https://doi.org/10.1038/nature13490

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Mantovani A, Marchesi F, Malesci A, Laghi L, Allavena P (2017) Tumour-associated macrophages as treatment targets in oncology. Nat Rev Clin Oncol 14(7):399–416. https://doi.org/10.1038/nrclinonc.2016.217

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Massague J (2008) TGFbeta in cancer. Cell 134(2):215–230. https://doi.org/10.1016/j.cell.2008.07.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Wrzesinski SH, Wan YY, Flavell RA (2007) Transforming growth factor-beta and the immune response: implications for anticancer therapy. Clin Cancer Res 13(18 Pt 1):5262–5270. https://doi.org/10.1158/1078-0432.Ccr-07-1157

    Article  CAS  PubMed  Google Scholar 

  36. Flavell RA, Sanjabi S, Wrzesinski SH, Licona-Limon P (2010) The polarization of immune cells in the tumour environment by TGFbeta. Nat Rev Immunol 10(8):554–567. https://doi.org/10.1038/nri2808

    Article  CAS  PubMed  Google Scholar 

  37. Moore KW, de Waal MR, Coffman RL, O'Garra A (2001) Interleukin-10 and the interleukin-10 receptor. Annu Rev Immunol 19:683–765. https://doi.org/10.1146/annurev.immunol.19.1.683

    Article  CAS  PubMed  Google Scholar 

  38. Fukumura D, Kloepper J, Amoozgar Z, Duda DG, Jain RK (2018) Enhancing cancer immunotherapy using antiangiogenics: opportunities and challenges. Nat Rev Clin Oncol 15(5):325–340. https://doi.org/10.1038/nrclinonc.2018.29

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Lo Russo G, Moro M, Sommariva M, Cancila V, Boeri M, Centonze G, Ferro S, Ganzinelli M, Gasparini P, Huber V, Milione M, Porcu L, Proto C, Pruneri G, Signorelli D, Sangaletti S, Sfondrini L, Storti C, Tassi E, Bardelli A, Marsoni S, Torri V, Tripodo C, Colombo MP, Anichini A, Rivoltini L, Balsari A, Sozzi G, Garassino MC (2019) Antibody-Fc/FcR interaction on macrophages as a mechanism for hyperprogressive disease in non-small cell lung cancer subsequent to PD-1/PD-L1 blockade. Clin Cancer Res 25(3):989–999. https://doi.org/10.1158/1078-0432.Ccr-18-1390

    Article  CAS  PubMed  Google Scholar 

  40. Gordon SR, Maute RL, Dulken BW, Hutter G, George BM, McCracken MN, Gupta R, Tsai JM, Sinha R, Corey D, Ring AM, Connolly AJ, Weissman IL (2017) PD-1 expression by tumour-associated macrophages inhibits phagocytosis and tumour immunity. Nature 545(7655):495–499. https://doi.org/10.1038/nature22396

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Funding

No relevant funding.

Author information

Authors and Affiliations

  1. Department of Gastrointestinal Tract Surgery, Fukushima Medical University School of Medicine, 1 Hikarigaoka, Fukushima, Fukushima, 960-1295, Japan

    Aung Kyi Thar Min, Kosaku Mimura, Shotaro Nakajima, Hirokazu Okayama, Katsuharu Saito, Wataru Sakamoto, Shotaro Fujita, Hisahito Endo, Motonobu Saito, Zenichiro Saze, Tomoyuki Momma, Shinji Ohki & Koji Kono

  2. Department of Blood Transfusion and Transplantation Immunology, Fukushima Medical University School of Medicine, 1 Hikarigaoka, Fukushima, Fukushima, 960-1295, Japan

    Kosaku Mimura

Authors
  1. Aung Kyi Thar Min
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kosaku Mimura
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shotaro Nakajima
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hirokazu Okayama
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Katsuharu Saito
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Wataru Sakamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Shotaro Fujita
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hisahito Endo
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Motonobu Saito
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Zenichiro Saze
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Tomoyuki Momma
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Shinji Ohki
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Koji Kono
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

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.

Corresponding author

Correspondence to Kosaku Mimura.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

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).

Informed consent

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.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00262-020-02676-8

Keywords

Search

Navigation