Phenotype screens of murine pancreatic cancer identify a Tgf-α-Ccl2-paxillin axis driving human-like neural invasion
Xiaobo Wang, … , Güralp Onur Ceyhan, Ihsan Ekin Demir
Xiaobo Wang, … , Güralp Onur Ceyhan, Ihsan Ekin Demir
Published August 22, 2023
Citation Information: J Clin Invest. 2023;133(21):e166333. https://doi.org/10.1172/JCI166333.
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Research Article Oncology

Phenotype screens of murine pancreatic cancer identify a Tgf-α-Ccl2-paxillin axis driving human-like neural invasion

Abstract

Solid cancers like pancreatic ductal adenocarcinoma (PDAC), a type of pancreatic cancer, frequently exploit nerves for rapid dissemination. This neural invasion (NI) is an independent prognostic factor in PDAC, but insufficiently modeled in genetically engineered mouse models (GEMM) of PDAC. Here, we systematically screened for human-like NI in Europe’s largest repository of GEMM of PDAC, comprising 295 different genotypes. This phenotype screen uncovered 2 GEMMs of PDAC with human-like NI, which are both characterized by pancreas-specific overexpression of transforming growth factor α (TGF-α) and conditional depletion of p53. Mechanistically, cancer-cell-derived TGF-α upregulated CCL2 secretion from sensory neurons, which induced hyperphosphorylation of the cytoskeletal protein paxillin via CCR4 on cancer cells. This activated the cancer migration machinery and filopodia formation toward neurons. Disrupting CCR4 or paxillin activity limited NI and dampened tumor size and tumor innervation. In human PDAC, phospho-paxillin and TGF-α–expression constituted strong prognostic factors. Therefore, we believe that the TGF-α-CCL2-CCR4-p-paxillin axis is a clinically actionable target for constraining NI and tumor progression in PDAC.

Authors

Xiaobo Wang, Rouzanna Istvanffy, Linhan Ye, Steffen Teller, Melanie Laschinger, Kalliope N. Diakopoulos, Kıvanç Görgülü, Qiaolin Li, Lei Ren, Carsten Jäger, Katja Steiger, Alexander Muckenhuber, Baiba Vilne, Kaan Çifcibaşı, Carmen Mota Reyes, Ümmügülsüm Yurteri, Maximilian Kießler, Ibrahim Halil Gürçınar, Maya Sugden, Saliha Elif Yıldızhan, Osman Uğur Sezerman, Sümeyye Çilingir, Güldal Süyen, Maximilian Reichert, Roland M. Schmid, Stefanie Bärthel, Rupert Oellinger, Achim Krüger, Roland Rad, Dieter Saur, Hana Algül, Helmut Friess, Marina Lesina, Güralp Onur Ceyhan, Ihsan Ekin Demir

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Figure 5

CCL2/CCR4 axis regulates paxillin phosphorylation and innervation in KPC mice.

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CCL2/CCR4 axis regulates paxillin phosphorylation and innervation in KPC...
(A) Experimental design of rCCL2 and C021 inhibitor treatment: 12-week-old KPC mice (n = 5) were injected i.p. with rCCL2 or C021 every other day for 3 weeks with normal saline as control. (B and C) Representative images of consecutive sections from PDAC samples of KPC mice treated with rCCL2, C021, and control groups stained with the neural marker PGP9.5 (pink), cancer cell marker CK-19 (brown), p-paxillin (brown), and counterstained with haematoxylin. Plots show (D) PGP9.5 content, (E) score of cancer cell proximity to neurons, and (F) p-paxillin content in PDAC sections from treated KPC mice. (G) The NI score in the pancreatic tumors of TPAC mice treated with the CCR4 inhibitor versus control (solvent) substance. All results in graphs are shown as a mean value ± SEM. For the statistical analyses, we used ordinary 1-way ANOVA (D–F), Dunnett’s multiple comparisons test (D–F) and for the distribution, a t test (G) and Shapiro-Wilk normality test (all panels). The P value ˂ 0.05 was considered to have significance. Scale bars: 20 μm.