Abstract
Purpose
This study aimed to demonstrate the involvement of angiogenesis in cancer-associated acinar-to-ductal metaplasia (CA-ADM) lesion of invasive front pancreatic ductal adenocarcinoma (PDAC) and investigate the possible mechanism.
Methods
Tissue samples from 128 patients with PDAC and 36 LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre mice were analyzed. Immunohistochemical assay was performed using HE, anti-CK19 and anti-amylase to confirm the presence of CA-ADM lesions, using anti-CD34 and anti-CD31 to measure microvessel density (MVD), and using anti-CD68, anti-CD163, anti-iNOS, or anti-MMP9 to evaluate the immune microenvironment. We performed multiplex immunohistochemical assay to detect the co-expression of MMP9 and CD68 on macrophage. We examined clinical outcomes and other clinicopathological factors to determine the significance of high-level MVD of CA-ADM on survival and liver metastasis. We performed tube formation assay to evaluate the effect of macrophage on angiogenic capacity in vitro.
Results
Angiogenesis was significantly abundant in CA-ADM lesions compared with that in PDAC lesions in human and mouse tissues. High-level MVD in CA-ADM lesions was an independent predictor of poor prognosis (P = 0.0047) and the recurrence of liver metastasis (P = 0.0027). More CD68-positive and CD163-positive macrophages were detected in CA-ADM lesions than in PDAC. The percentage of CD68-positive macrophages was positively correlated with MVD in CA-ADM lesions. Multiplex-immunostaining revealed that MMP9 was expressed in CD68-positive macrophages of CA-ADM lesions. In CA-ADM lesions, the percentage of macrophages was positively correlated with MMP9 expression, which positively correlated with microvessel density.
Conclusion
CA-ADM related angiogenesis is a promising predictive marker for poor prognosis of PDAC and may provide an attractive therapeutic target for PDAC.
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Data availability statement
The data presented in this research are available from the corresponding author upon request.
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Acknowledgements
We appreciate the technical support provided by Emiko Manabe and Shoko Sadatomi (Department of Surgery and Oncology, Kyushu University). Shuang Fei is the recipient of Otsuka Toshimi (2021-2022) [http://www.otsukafoundation.org/index.html] and Rotary Yoneyama Memorial Foundation scholarship (2022-2023) [http://www.rotary-yoneyama.or.jp]. We thank Gabrielle White Wolf, PhD, from Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript.
Funding
This work was supported by Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Numbers JP19K22663, JP19K22664, JP22H02922, JP22K15560, JP22K16490.
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Shuang Fei: conceptualization, methodology, formal analysis, investigation, project administration, writing review and editing. Kenoki Ohuchida: conceptualization, investigation, methodology, project administration, writing review and editing. Shin Kibe and Zilong Yan: conceptualization and investigation. Chika Iwamoto and Shinkawa Tomohiko: methodology. Bo Zhang: data curation. Jun Kawata: resources and investigation. Toshiya Abe and Noboru Ideno: investigation. Naoki Ikenaga: formal analysis. Kohei Nakata: methodology. Yoshinao Oda: conceptualization and resources. Masafumi Nakamura: project administration, supervision writing review and editing. The first draft of the manuscript was written by Shuang Fei and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Fei, S., Ohuchida, K., Kibe, S. et al. Involvement of angiogenesis in cancer-associated acinar-to-ductal metaplasia lesion of pancreatic cancer invasive front. J Cancer Res Clin Oncol 149, 5885–5899 (2023). https://doi.org/10.1007/s00432-022-04554-5
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DOI: https://doi.org/10.1007/s00432-022-04554-5