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 1

Mutant TPAC mice represent a model for human-like PNI.

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Mutant TPAC mice represent a model for human-like PNI. Representative im...
Representative images of genuine PNI in PDAC resection specimens (A) and in TPAC mouse mutants (B, please also see Supplemental Figure 1E for a broader view of the tumor area with PNI); pan-neural marker: PGP9.5 (brown), cancer cell marker: CK-19 (pink), and Aniline blue (for collagen). (C) Graphs showing the severity of NI in GEMMs of PDAC. (D) Scheme of migration assay of cancer cells and neurons from dorsal root ganglia (DRGs). (E) Graphs show the FMI of primary cancer cells isolated from KC, KPC, TPC, and TPAC mice in the 3D migration assay. (F) Representative images of DRG neurons treated with conditioned medium from KPC, TPC, and TPAC cancer cells, stained with neuronal marker β3 Tubulin (Tubβ3) antibody. (G) Results of the in vitro neuroplasticity assay of DRG neurons treated with supernatants of the primary cancer cells from the analyzed mouse genotypes. (H) Transcriptome analysis of TPAC- versus KPC-derived primary cancer cells. (I) Schematic representation of the 3D SC outgrowth assay. Sciatic nerves isolated from WT mice were placed between bridges connected to ECM gel drops containing primary murine cancer cells (KC, KPC, TPC, and TPAC) and empty ECM gel drops. The test was performed for 72 hours in culture media with CO2 supply. (J) Graphs showing the FMI of SCs. All results in the graphs are shown as mean ± SEM. For statistical analyses, Mann-Whitney U test (C and E), ordinary 1-way ANOVA (G), Kruskal-Wallis test, Dunnett’s test for multiple comparisons (G and H) and Shapiro-Wilk normality test (all panels) for distribution were used.