TIMP1 Triggers Neutrophil Extracellular Trap Formation in Pancreatic Cancer

Abstract

Tumor-derived protein tissue inhibitor of metalloproteinases-1 (TIMP1) correlates with poor prognosis in many cancers, including highly lethal pancreatic ductal adenocarcinoma (PDAC). The noncanonical signaling activity of TIMP1 is emerging as one basis for its contribution to cancer progression. However, TIMP1–triggered progression-related biological processes are largely unknown. Formation of neutrophil extracellular traps (NET) in the tumor microenvironment is known to drive progression of PDAC, but factors or molecular mechanisms initiating NET formation in PDAC remain elusive. In this study, gene-set enrichment analysis of a human PDAC proteome dataset revealed that TIMP1 protein expression most prominently correlates with neutrophil activation in patient-derived tumor tissues. TIMP1 directly triggered formation of NETs in primary human neutrophils, which was dependent on the interaction of TIMP1 with its receptor CD63 and subsequent ERK signaling. In genetically engineered PDAC-bearing mice, TIMP1 significantly contributed to NET formation in tumors, and abrogation of TIMP1 or NETs prolonged survival. In patient-derived PDAC tumors, NETs predominantly colocalized with areas of elevated TIMP1 expression. Furthermore, TIMP1 plasma levels correlated with DNA-bound myeloperoxidase, a NET marker, in the blood of patients with PDAC. A combination of plasma levels of TIMP1 and NETs with the clinically established marker CA19–9 allowed improved identification of prognostically distinct PDAC patient subgroups. These observations may have a broader impact, because elevated systemic levels of TIMP1 are associated with the progression of a wide range of neutrophil-involved inflammatory diseases. Significance: These findings highlight the prognostic relevance of TIMP1 and neutrophil extracellular traps in highly lethal pancreatic cancer, where a noncanonical TIMP1/CD63/ERK signaling axis induces NET formation.

Neutrophil in the Pancreatic Tumor Microenvironment

“…Tumor-derived protein tissue inhibitor of metalloproteinases-1 (TIMP-1), which correlated with poor prognosis in PDAC, directly triggered the formation of NETs. This effect depended on the interaction of TIMP-1 with its receptor CD63 and subsequent ERK signaling [69]. In addition to tumor cells, CAF-conditioned media can also induce ROS-dependent NETosis by secreting Amyloid β A4 protein.…”

“…A recent study showed that plasma NET levels using SYTOX-positive areas could also predict the survival of PDAC patients [69]. High plasma levels of NET markers were indicators of increased death risk of PDAC patients.…”

PEGylation of Metal Oxide Nanoparticles Modulates Neutrophil Extracellular Trap Formation

“…Although not conventionally thought to be released by neutrophils themselves, some studies have illustrated that neutrophils do in fact produce SDF-1 at least at the mRNA level [ 127 ]. TIMP-1 is also known to elicit NETosis, particularly in the context of pancreatic cancer [ 128 ]. The production of each cytokine, but lack of increased NETosis, suggests that GBCAs have the ability to influence neutrophil chemoattraction.…”

Neutrophil Extracellular Traps in Digestive Cancers: Warrior or Accomplice

“…However, reports have shown that endogenous G-CSF promotes the progress of several types of cancers, such as neuroblastoma and breast cancer (56,57), and tumor-derived G-CSF plays a critical role in NET formation in breast cancer and lung cancer (39,41,58). Tumor cells can also induce NET formation by secreting a variety of other stimulating factors, such as exosome, transforming growth factor-b (TGF-b), interleukin-1b (IL-1b), extracellular RNA (exRNA), mitochondrial DNA (mtDNA), high mobility group box-1 (HMGB1), cathepsin C (CTSC), tissue inhibitor of metalloproteinases-1 (TIMP-1), CXCL5 (59)(60)(61)(62)(63)(64)(65)(66)(67)(68)(69).…”

“…Abnormally increased NETs are observed in pancreatic cancer patients and the models of murine orthotopic pancreatic cancer (95,172); these NETs can be regarded as an independent prognostic factor for evaluating the outcomes of patients with PDAC, which are negatively correlated with the OS and recurrence-free survival of patients (40). Existing studies showed that increased levels of NETs in pancreatic cancer are partly due to the direct effect of tumor cells, cancer-associated fibroblasts, or activated platelet (67,76,79,106).…”