Tissue-specific reprogramming leads to angiogenic neutrophil specialization and tumor vascularization in colorectal cancer
Triet M. Bui, … , Stephen B. Hanauer, Ronen Sumagin
Triet M. Bui, … , Stephen B. Hanauer, Ronen Sumagin
Published February 8, 2024
Citation Information: J Clin Invest. 2024;134(7):e174545. https://doi.org/10.1172/JCI174545.
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Tissue-specific reprogramming leads to angiogenic neutrophil specialization and tumor vascularization in colorectal cancer

Abstract

Neutrophil (PMN) tissue accumulation is an established feature of ulcerative colitis (UC) lesions and colorectal cancer (CRC). To assess the PMN phenotypic and functional diversification during the transition from inflammatory ulceration to CRC we analyzed the transcriptomic landscape of blood and tissue PMNs. Transcriptional programs effectively separated PMNs based on their proximity to peripheral blood, inflamed colon, and tumors. In silico pathway overrepresentation analysis, protein-network mapping, gene signature identification, and gene-ontology scoring revealed unique enrichment of angiogenic and vasculature development pathways in tumor-associated neutrophils (TANs). Functional studies utilizing ex vivo cultures, colitis-induced murine CRC, and patient-derived xenograft models demonstrated a critical role for TANs in promoting tumor vascularization. Spp1 (OPN) and Mmp14 (MT1-MMP) were identified by unbiased -omics and mechanistic studies to be highly induced in TANs, acting to critically regulate endothelial cell chemotaxis and branching. TCGA data set and clinical specimens confirmed enrichment of SPP1 and MMP14 in high-grade CRC but not in patients with UC. Pharmacological inhibition of TAN trafficking or MMP14 activity effectively reduced tumor vascular density, leading to CRC regression. Our findings demonstrate a niche-directed PMN functional specialization and identify TAN contributions to tumor vascularization, delineating what we believe to be a new therapeutic framework for CRC treatment focused on TAN angiogenic properties.

Authors

Triet M. Bui, Lenore K. Yalom, Edward Ning, Jessica M. Urbanczyk, Xingsheng Ren, Caroline J. Herrnreiter, Jackson A. Disario, Brian Wray, Matthew J. Schipma, Yuri S. Velichko, David P. Sullivan, Kouki Abe, Shannon M. Lauberth, Guang-Yu Yang, Parambir S. Dulai, Stephen B. Hanauer, Ronen Sumagin

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Figure 1

Tissue compartmentalization of PMN immunophenotypes during colitis-to-CRC transition.

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Tissue compartmentalization of PMN immunophenotypes during colitis-to-CR...
(A) Endoscopic (left) and histological images (right) of healthy, ulcerated and CRC-bearing colons induced by AOM/DSS treatment. Acute colitis 1 DSS cycle, early and advanced CRC with 3 and 5 DSS cycles, respectively. Images representative of n = 4 independent experiments. Scale bars: 1 mm for endoscopy and 100 μm for H&E. (B) IHC staining and (C) quantification of S100A9 PMNs in healthy, inflamed colitis and AOM/DSS induced tumors (n = 3–4 mice per condition, with approximately 50 fields of view [FOVs]per condition). Insets are higher magnification images showing PMN accumulation in tumors. Scale bar: 50 μm. (D) Whole-mount confocal imaging of AOM/DSS-induced CRC tumors in Lyz2EGFP reporter mice. ECs were visualized by PECAM-1 and tumor cells by EpCAM staining. The inset shows magnified image of area highlighted by dotted line, depicting PMN interacting with blood vessels within the tumor niche. Scale bar: 50 μm. Images representative of n = 4 independent experiments. (E) Flow cytometric analyses of Ly6G+/CD11b+/Lyz2EGFP PMN numbers across tissue compartments during CRC development. n = 5–8 for healthy and colitis and n = 8–12 for CRC. Data are shown as mean ± SEM. **P < 0.01, ***P < 0.001, significance between diseased conditions; ##P < 0.01, significance between cancer and adjacent tissue(1-way ANOVA with Tukey’s multiple comparison test). (F) 3D PCA matrix of RNA-Seq profiles of 7 spatiotemporal conditions. 3 separate PMN clusters (dotted circles) were identified, representing peripheral blood, inflamed colon, and CRC niches.