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Muc4 loss mitigates epidermal growth factor receptor activity essential for PDAC tumorigenesis

Oncogene volume 42, pages 759–770 (2023)Cite this article

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Abstract

Mucin4 (MUC4) appears early during pancreatic intraepithelial neoplasia-1 (PanIN1), coinciding with the expression of epidermal growth factor receptor-1 (EGFR). The EGFR signaling is required for the onset of Kras-driven pancreatic ductal adenocarcinoma (PDAC); however, the players and mechanisms involved in sustained EGFR signaling in early PanIN lesions remain elusive. We generated a unique Esai-CRISPR-based Muc4 conditional knockout murine model to evaluate its effect on PDAC pathology. The Muc4 depletion in the autochthonous murine model carrying K-ras and p53 mutations (K-rasG12D; TP53R172H; Pdx-1cre, KPC) to generate the KPCM4−/− murine model showed a significant delay in the PanIN lesion formation with a significant reduction (p < 0.01) in EGFR (Y1068) and ERK1/2 (T202/Y204) phosphorylation. Further, a significant decrease (p < 0.01) in Sox9 expression in PanIN lesions of KPCM4−/− mice suggested the impairment of acinar-to-ductal metaplasia in Muc4-depleted cells. The biochemical analyses demonstrated that MUC4, through its juxtamembrane EGF-like domains, interacts with the EGFR ectodomain, and its cytoplasmic tail prevents EGFR ubiquitination and subsequent proteasomal degradation upon ligand stimulation, leading to sustained downstream oncogenic signaling. Targeting the MUC4 and EGFR interacting interface provides a promising strategy to improve the efficacy of EGFR-targeted therapies in PDAC and other MUC4-expressing malignancies.

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Fig. 1: The Muc4 depletion delays the initiation of PDAC.
Fig. 2: The early lesions in the pancreatic tissues of KPCM4−/− mice display significantly lower EGFR and ERK1/2 phosphorylation status.
Fig. 3: Muc4 depletion significantly reduces acinar-to-ductal metaplasia (ADM) in murine pancreatic tissues.
Fig. 4: Muc4 expression shows a significant correlation with EGFR activity in the differentiating ductal lesions in the pancreatic tissues.
Fig. 5: MUC4 interacts with EGFR upon ligand stimulation leading to sustained signaling.
Fig. 6: MUC4 expression maintains EGFR activity by preventing its ubiquitination upon ligand stimulation.

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All data needed to evaluate the conclusions in the paper are present in the manuscript and the Supplementary Materials.

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Acknowledgements

We sincerely thank Dr. Imayavaramban Lakshmanan for providing Her2 silenced CD18/HPAF cell line. We thank Jessica Mercer for editing the manuscript. We thank the UNMC Department of Pathology for providing the patient tissue samples. We thank Saswati Karmakar and Ramesh Pothuraju for helping us in characterizing Muc4 COIN mice.

Funding

The authors/work was partly supported by funding from the National Institutes of Health (P01 CA217798, U01 CA210240, U01 CA200466, R01 CA206444, R21 AA026428, R01 CA228524, R01 CA247471, R01 CA254036, R01 CA256973, R01 CA263575, R01 CA273349, R01 CA218545, R01 CA241752, and R44 CA235991), and W81XWH-21-1-0640.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE, USA

    Rakesh Bhatia, Jawed Akhtar Siddiqui, Koelina Ganguly, Christopher M. Thompson, Andrew Cannon, Abhijit Aithal, Naveenkumar Perumal, Shailendra K. Maurya, Xiaoqi Li, Satyanarayana Rachagani, Maneesh Jain, Mohd Wasim Nasser, Surinder K. Batra & Sushil Kumar

  2. Fred and Pamela Buffett Cancer Center, Omaha, NE, USA

    Jawed Akhtar Siddiqui, Maneesh Jain, Mohd Wasim Nasser, Surinder K. Batra & Sushil Kumar

  3. Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE, USA

    Jesse L. Cox

  4. Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE, USA

    Channabasavaiah B. Gurumurthy

  5. Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE, USA

    Surinder K. Batra

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  1. Rakesh Bhatia
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Contributions

RB performed biochemical analysis and wrote the manuscript. RB, JAS, NP, SKM, and XL generated and characterized KPCM4−/− murine model. KG performed I.H.C. experiments. CMT and AC performed bioinformatics analysis. AA generated the MUC4 monoclonal antibody. JLC scored I.H.C. slides. CBG helped in the generation of Muc4 COIN mice. SR, MJ, and MWN provided inputs in murine model studies. SK conceived the idea and designed the experiments. SK and SKB supervised the project and approved the manuscript.

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Correspondence to Surinder K. Batra or Sushil Kumar.

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SKB is a founder of Sanguine Diagnostics and Therapeutics, Inc. Other authors have no competing interests to declare.

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Bhatia, R., Siddiqui, J.A., Ganguly, K. et al. Muc4 loss mitigates epidermal growth factor receptor activity essential for PDAC tumorigenesis. Oncogene 42, 759–770 (2023). https://doi.org/10.1038/s41388-022-02587-1

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