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Cellular and Molecular Biology

Neuropeptide Y, a paracrine factor secreted by cancer cells, is an independent regulator of angiogenesis in colon cancer

Abstract

Background

Resistance to anti-angiogenic therapies targeting vascular endothelial growth factor-A (VEGF-A) stems from VEGF-A independent angiogenesis mediated by other proangiogenic factors. Therefore identifying these factors in colon adenocarcinoma (CA) will reveal new therapeutic targets.

Methods

Neuropeptide Y (NPY) and Y2 receptor (Y2R) expressions in CA were studied by immunohistochemical analysis. Orthotopic HT29 with intact VEGF-A gene and VEGF-A knockdown (by CRISPR/Cas9 gene-editing technique) HT29 colon cancer-bearing mice were treated with specific Y2R antagonists, and the effects on angiogenesis and tumour growth were studied. The direct effect of NPY on angiogenesis and the underlying molecular mechanism was elucidated by the modulation of Y2R receptors expressed on colonic endothelial cells (CEC).

Results

The results demonstrated that NPY and Y2R are overexpressed in human CA, orthotopic HT29, and most interestingly in VEGF-A-depleted orthotopic HT29 tumours. Treatment with Y2R antagonists inhibited angiogenesis and thereby HT29 tumour growth. Blocking /silencing Y2R abrogated NPY-induced angiogenic potential of CEC. Mechanistically, NPY regulated the activation of the ERK/MAPK signalling pathway in CEC.

Conclusions

NPY derived from cancer cells independently regulates angiogenesis in CA by acting through Y2R present on CEC. Targeting NPY/Y2R thus emerges as a novel potential therapeutic strategy in CA.

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Fig. 1: Expressions of neuropeptide Y (NPY) and its receptor (Y2R) are significantly upregulated in colon adenocarcinoma (CA).
Fig. 2: Neuropeptide Y (NPY) and Y2R are overexpressed in orthotopic HT29 colon cancer.
Fig. 3: Significant upregulation of angiogenesis (CD31 + ve vessels) in colon adenocarcinoma shows a strong positive correlation with NPY expression.
Fig. 4: Significant tumour growth and angiogenesis inhibition upon treatment of orthotopic HT29 and HT29 VEGF-A knockout (VEGF-A KO HT29) tumour-bearing mice with Y2R antagonists, BIIE0246 and JNJ5207787.
Fig. 5: NPY stimulates the angiogenic potential of colonic endothelial cells via Y2R expressed on these cells.
Fig. 6: NPY stimulates angiogenesis in colon cancer by activating the ERK/MAPK pathway.

Data availability

All data relevant to the study are included in the article or uploaded as supplementary information.

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Acknowledgements

We acknowledge the support of the veterinarian and vivarium staff of OSU ULAR for our animal studies and OSU Pathology Core facilities for confocal microscopy studies.

Funding

This study was supported by funding from NIH/NCI [R21 CA216763 to CS]. CS and DC would like to acknowledge the funding received from Mitchell Cancer Institute, University of South Alabama. DC was also supported by DOD, USA [W81XWH-20-1-0618], and SB was supported by DOD, USA [W81XWH-19-1-0233 and W81XWH2110874].

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Conception: CS; design: CS, DC and SG; development of methodology: DC, SG and CS; experiment and data generation: DC, SG and HF; analysis and interpretation of data: CS, DC, SG, HF, WF and SB; writing original draft: DC, SG and CS; writing/reviewing and editing: DC, SG, HF, WF, SB and CS; administrative and study supervision: DC and CS. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Chandrani Sarkar.

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Only de-identified and archived human colon cancer tissues were used under an exempt (category 4) protocol as determined by The Ohio State University Institutional Review Board. The study was performed in accordance with the Declaration of Helsinki.

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Chakroborty, D., Goswami, S., Fan, H. et al. Neuropeptide Y, a paracrine factor secreted by cancer cells, is an independent regulator of angiogenesis in colon cancer. Br J Cancer (2022). https://doi.org/10.1038/s41416-022-01916-1

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