Macrophages direct cancer cells through a LOXL2-mediated metastatic cascade in pancreatic ductal adenocarcinoma.

Abstract

The lysyl oxidase-like protein 2 (LOXL2) contributes to tumour progression and metastasis in different tumour entities, but its role in pancreatic ductal adenocarcinoma (PDAC) has not been evaluated in immunocompetent in vivo PDAC models. Towards this end, we used PDAC patient data sets, patient-derived xenograft in vivo and in vitro models, and four conditional genetically-engineered mouse models (GEMMS) to dissect the role of LOXL2 in PDAC. For GEMM-based studies, <i>K-Ras</i> ^+/LSL-G12D;<i>Trp53</i> ^LSL-R172H;<i>Pdx1-Cre</i> mice (KPC) and the <i>K-Ras</i> ^+/LSL-G12D;<i>Pdx1-Cre</i> mice (KC) were crossed with <i>Loxl2</i> allele floxed mice (<i>Loxl2^Exon2</i> ^fl/fl) or conditional <i>Loxl2</i> overexpressing mice (R26<i>Loxl2</i> ^KI/KI) to generate KPCL2^KO or KCL2^KO and KPCL2^KI or KCL2^KI mice, which were used to study overall survival; tumour incidence, burden and differentiation; metastases; epithelial to mesenchymal transition (EMT); stemness and extracellular collagen matrix (ECM) organisation. Using these PDAC mouse models, we show that while <i>Loxl2</i> ablation had little effect on primary tumour development and growth, its loss significantly decreased metastasis and increased overall survival. We attribute this effect to non-cell autonomous factors, primarily ECM remodelling. <i>Loxl2</i> overexpression, on the other hand, promoted primary and metastatic tumour growth and decreased overall survival, which could be linked to increased EMT and stemness. We also identified tumour-associated macrophage-secreted oncostatin M (OSM) as an inducer of LOXL2 expression, and show that targeting macrophages in vivo affects <i>Osm</i> and <i>Loxl2</i> expression and collagen fibre alignment. Taken together, our findings establish novel pathophysiological roles and functions for LOXL2 in PDAC, which could be potentially exploited to treat metastatic disease.