Annexin A2/TLR2/MYD88 pathway induces arginase 1 expression in tumor-associated neutrophils
Huajia Zhang, Xiaodong Zhu, Travis J. Friesen, Jeff W. Kwak, Tatyana Pisarenko, Surapat Mekvanich, Mark A. Velasco, Timothy W. Randolph, Julia Kargl, A. McGarry Houghton
Huajia Zhang, Xiaodong Zhu, Travis J. Friesen, Jeff W. Kwak, Tatyana Pisarenko, Surapat Mekvanich, Mark A. Velasco, Timothy W. Randolph, Julia Kargl, A. McGarry Houghton
View: Text | PDF
Research Article Immunology Oncology

Annexin A2/TLR2/MYD88 pathway induces arginase 1 expression in tumor-associated neutrophils

Abstract

Myeloid lineage cells suppress T cell viability through arginine depletion via arginase 1 (ARG1). Despite numerous studies exploring the mechanisms by which ARG1 perturbs lymphocyte function, the cellular populations responsible for its generation and release remain poorly understood. Here, we showed that neutrophil lineage cells and not monocytes or macrophages expressed ARG1 in human non–small cell lung cancer (NSCLC). Importantly, we showed that approximately 40% of tumor-associated neutrophils (TANs) actively transcribed ARG1 mRNA. To determine the mechanism by which ARG1 mRNA is induced in TANs, we utilized FPLC followed by MS/MS to screen tumor-derived factors capable of inducing ARG1 mRNA expression in neutrophils. These studies identified ANXA2 as the major driver of ARG1 mRNA expression in TANs. Mechanistically, ANXA2 signaled through the TLR2/MYD88 axis in neutrophils to induce ARG1 mRNA expression. The current study describes what we believe to be a novel mechanism by which ARG1 mRNA expression is regulated in neutrophils in cancer and highlights the central role that neutrophil lineage cells play in the suppression of tumor-infiltrating lymphocytes.

Authors

Huajia Zhang, Xiaodong Zhu, Travis J. Friesen, Jeff W. Kwak, Tatyana Pisarenko, Surapat Mekvanich, Mark A. Velasco, Timothy W. Randolph, Julia Kargl, A. McGarry Houghton

×

Figure 7

ANXA2 induces ARG1 gene expression in human neutrophils.

Options: View larger image (or click on image) Download as PowerPoint
ANXA2 induces ARG1 gene expression in human neutrophils. (A) Representat...
(A) Representative images from FFPE NSCLC slides (n = 6) stained for CD68/CD163 (yellow), ANXA2 (purple), and CK (white). Images farthest to the left are ×20 original magnification, all others are ×40. (B) Representative M-IHC and spatial plot images for ANXA2 (purple) staining and CD66b (green), ARG1 IHC (yellow), and ARG1-FISH (red) staining. (C) Percentage of ANXA2+ cells also staining positively for CK, CD68/CD163, or neither (other). Bars denote ± SEM. (D) Tabulation of ARG1-FISH+ cells as a function of location either inside or outside the malignant tumor boundary. n = 12. Bars denote ± SEM. HDNs from healthy donors were isolated from peripheral blood within 1 hour after blood draw. HDNs were incubated with (E) human NSCLC tumor lysate (100 μg/mL) or (F) recombinant human ANXA2 for 1 hour. Results expressed as fold change of ARG1 mRNA expression in the treatment group compared with PBS control. PBS, n = 6; tumor lysate, n = 14. ANXA2 experiments from a representative experiment in triplicate. (G) ANXA2 ELISA from human NSCLC tumor lysates (n = 9). Results expressed as ANXA2 protein concentration in ng/mL. (H) HDNs from healthy donors as in E incubated with IL-10, IL-4, or LTA for 1 hour. Results expressed as fold change in ARG1 gene expression. n = 4 per group. *P < 0.05; ***P < 0.001 by 1-way ANOVA with Tukey’s post hoc test (C and H) or 2-tailed Student’s t test (D–F). (I) Monocyte-derived macrophages were generated from human peripheral blood and incubated with IL-10 (10 U/mL), IL-4 (10 U/mL), or LTA (20 μg/mL) for 1 hour. Results expressed as fold change in ARG1 gene expression. n = 4 per group. P > 0.05 by 1-way ANOVA with Tukey’s post hoc test.