Tissue-specific reprogramming leads to angiogenic neutrophil specialization and tumor vascularization in colorectal cancer
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
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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Research Article Gastroenterology Immunology

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 4

TANs promote tumor vascularization and vessel-tumor depth penetration.

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TANs promote tumor vascularization and vessel-tumor depth penetration. (...
(A) Representative IHC images of intratumoral (green) and stromal (blue) blood vessels (stained for PECAM-1) in PMNhi (isotype-treated) and PMNlo (treated with anti-Ly6G/MAR18.5) tumors. Scale bar: 200 μm. (B) High-power IHC images depict elevated vascular density in PMNhi versus PMNlo tumors. Scale bar: 50 μm. Dashed line demarcates intratumor versus stromal regions. (C) Quantification of vessel density from images shown in A and B (n = 5 mice and 8–13 whole-tumor sections per condition). (D) Quantification of vessel-tumor depth penetration and TAN presence. (E) 3D reconstruction (rendering planes in XZ direction) and Z-stack projections of whole-mount tumor tissue in early and advanced PMNhi versus PMNlo tumors. For each image, 100 μm stacks were generated using 1 μm-focal depth steps. (F) Orthogonal views used for vessel depth penetration analyses (right) depicting a tricellular contact between TANs (green), invading blood vessels (blue), and the CRC interface (red) (n = 4–6 mice per condition with 50–70 z-stacked images analyzed). Scale bar 10 μm. *P < 0.05, **P < 0.01, ****P < 0.0001, 1-way ANOVA with Tukey’s multiple comparison test.