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. 2020 Mar 16;11(3):187.
doi: 10.1038/s41419-020-2371-x.

PRDM3 attenuates pancreatitis and pancreatic tumorigenesis by regulating inflammatory response

Jie Ye #  1   2 , Anpei Huang #  3 , Haitao Wang  1   4 , Anni M Y Zhang  5 , Xiaojun Huang  1   2 , Qingping Lan  1   2 , Tomohiko Sato  6 , Susumu Goyama  6 , Mineo Kurokawa  6 , Chuxia Deng  1   2 , Maike Sander  7 , David F Schaeffer  8 , Wen Li  9 , Janel L Kopp  10 , Ruiyu Xie  11   12
Affiliations

PRDM3 attenuates pancreatitis and pancreatic tumorigenesis by regulating inflammatory response

Jie Ye et al. Cell Death Dis. .

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is associated with metaplastic changes in the pancreas but the transcriptional program underlying these changes is incompletely understood. The zinc finger transcription factor, PRDM3, is lowly expressed in normal pancreatic acini and its expression increases during tumorigenesis. Although PRDM3 promotes proliferation and migration of PDAC cell lines, the role of PRDM3 during tumor initiation from pancreatic acinar cells in vivo is unclear. In this study, we showed that high levels of PRDM3 expression in human pancreas was associated with pancreatitis, and well-differentiated but not poorly differentiated carcinoma. We examined PRDM3 function in pancreatic acinar cells during tumor formation and pancreatitis by inactivating Prdm3 using a conditional allele (Ptf1aCreER;Prdm3flox/flox mice) in the context of oncogenic Kras expression and supraphysiological cerulein injections, respectively. In Prdm3-deficient mice, KrasG12D-driven preneoplastic lesions were more abundant and progressed to high-grade precancerous lesions more rapidly. This is consistent with our observations that low levels of PRDM3 in human PDAC was correlated significantly with poorer survival in patient. Moreover, loss of Prdm3 in acinar cells elevated exocrine injury, enhanced immune cell activation and infiltration, and greatly increased acinar-to-ductal cell reprogramming upon cerulein-induced pancreatitis. Whole transcriptome analyses of Prdm3 knockout acini revealed that pathways involved in inflammatory response and Hif-1 signaling were significantly upregulated in Prdm3-depleted acinar cells. Taken together, our results suggest that Prdm3 favors the maintenance of acinar cell homeostasis through modulation of their response to inflammation and oncogenic Kras activation, and thus plays a previously unexpected suppressive role during PDAC initiation.

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Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

Fig. 1
Fig. 1. Weak PRDM3 expression is associated with poorly differentiated tumors and a worse prognostic outcome in patients with PDAC.
Prdm3 immunostaining in normal pancreatic tissue (a and a’), pancreatitis (b and b’), PanIN (c and c’), well-differentiated (d and d’), moderately differentiated (e and e’), and poorly differentiated (f and f’) PDAC. g The overall survival probability and disease-free survival probability were compared between low (n = 32, staining index ≤ 6) and high (n = 62, staining index > 6) levels of PRDM3 expression in a cohort of 94 PDAC patients after surgical resection. p-values were calculated based on log-rank test. Scale: 50 μm.
Fig. 2
Fig. 2. Loss of prdm3 promotes acinar-to-ductal metaplasia and PanIN lesions formation.
a Ptf1aCreER;KrasG12D and Ptf1aCreER;KrasG12D;Prdm3flox/flox mice at 4–5 weeks of age were injected 4 times on alternating days with tamoxifen. Recombined mice were analyzed at 4 and 6 weeks post tamoxifen injection. b The number of PanINs per section for KrasG12D (n = 5) vs. KrasG12D-Prdm3ΔAcinar (n = 5) mice at 4 weeks post tamoxifen injection. c Representative images of high-grade PanINs in KrasG12D-Prdm3ΔAcinar mice at 4 weeks after tamoxifen injection. d Hematoxylin-eosin staining (H&E) and immunohistochemistry staining for the ductal marker Cytokeratin 19 (CK19). Quantification of the number of PanINs, as well as the percent of pancreatic area that is CK19+ in KrasG12D (n = 9) vs. KrasG12D-Prdm3ΔAcinar (n = 9) mice 6 weeks post-tamoxifen injection. e Images of acinar cell explants embedded in Matrigel at 24 and 48 h. f Quantification of the percent of ductal-like structures in explants derived from KrasG12D (n = 3) and KrasG12D-Prdm3ΔAcinar (n = 3) mice at 4 weeks post-tamoxifen injection. Data show mean ± SD. Statistical analysis: Two-tailed t-test. *p < 0.05, **p < 0.01, ***p < 0.001. Scale: 100 μm.
Fig. 3
Fig. 3. Inhibition of Prdm3 accelerates KrasG12D-driven neoplastic transformation in response to pancreatitis.
a Schematic illustration showing experimental design of cerulein-induced acute pancreatitis in cooperation with activation of oncogenic Kras in Ptf1a-expressing cells. Ptf1aCreER; KrasG12D (n = 7) and Ptf1aCreER; KrasG12D; Prdm3flox/flox (n = 7) mice at 4 to 5 weeks of age were injected 4 times on alternating days with tamoxifen. One week after the last tamoxifen injection, mice were subjected to cerulein (50 μg/kg) injection at hourly intervals over 6 h on alternating days separated by 24 h and analyzed at 21 days post cerulein injection. b Quantification of relative pancreas mass measured as percent of pancreas weight over body weight in KrasG12D vs. KrasG12D-Prdm3ΔAcinar mice. c Hematoxylin-eosin staining (H&E) and immunohistochemistry for Cpa1, Cytokeratin 19 (CK19) and Alcian blue of pancreata from KrasG12D and KrasG12D-Prdm3ΔAcinar mice. Quantification of the percent of pancreatic area that is Cpa1+, CK19+ or Alcian blue+ in KrasG12D vs. KrasG12D-Prdm3ΔAcinar mice. d Representative images of tumor budding in KrasG12D-Prdm3ΔAcinar mice indicated by red arrowheads. Immunohistochemistry staining of CK19 strongly suggests invasive high-grade neoplasia. e Number of high-grade PanINs per section for each genotype. Data show mean ± SD. Statistical analysis: Two-tailed t-test. *p < 0.05, **p < 0.01, ***p < 0.001. Scale: 200 μm (c) and 100 μm (d).
Fig. 4
Fig. 4. Prdm3 deletion in pancreatic acinar cells exaggerates cerulein-induced pancreatitis.
a Schematic illustration of experimental design. Ptf1aCreER;Prdm3flox/flox mice at 4 to 5 weeks of age were injected 4 times on alternating days with tamoxifen. One week after the last tamoxifen injection, mice were subjected to cerulein (50 μg/kg) injection at hourly intervals for 8 h per day for two consecutive days and analyzed at the indicated time points. b Histologic characterization was determined with hematoxylin-eosin staining (H&E). Immunohistochemistry staining for neutrophil marker Ly6B.2 and macrophage marker F4/80 with quantitation of the percent of all cells that are Ly6B.2+ or F4/80+ in Ptf1aCreER (control, n = 7) and Prdm3ΔAcinar (n = 5) pancreata. Vacuole is indicated by yellow arrowheads, and infiltrated immune cells are indicated by red arrowheads. c Serum amylase levels of cerulein-treated control (n = 11) vs. Prdm3ΔAcinar (n = 11) mice and saline-treated control (n = 3) vs. Prdm3ΔAcinar (n = 3). d Representative images of control and Prdm3ΔAcinar pancreata stained with H&E and the ductal marker Cytokeratin 19 (CK19). Quantification of the respective percent of all cells that are CK19+ from control (n = 6) vs. Prdm3ΔAcinar (n = 5) pancreata. Data show mean ± SD. Statistical analysis: Two-tailed t-test. *p < 0.05. Scale: 100 μm.
Fig. 5
Fig. 5. Inactivation of Prdm3 enhances inflammatory response pathways.
a Cytokine secretion was determined using a Raybiotech mouse inflammation array. Primary acinar cells, isolated from control (n = 3) and Prdm3ΔAcinar (n = 3) mice, were exposed to cerulein (10 nM) or TNF-α (100 ng/ml) for 6 h. Levels of cytokines (MIP-1α and RANTES) in culture medium were expressed relative to control. p-values were calculated based on two-tailed t-test. b Primary acinar cells were isolated from Prf1aCreER (n = 5) vs. Prdm3ΔAcinar (n = 5) mice and cultured in Waymouth complete media in the presence of 10 nM cerulein for 6 h. Raw264.7 cells were treated with the above acinar-cell-conditioned media for 16 h and collected for quantitative real-time PCR for the indicated cytokines. Each set of connected dots in b depicts an independent biological replicate (n = 5 experiments). Data show mean ± SD. Statistical analysis: Two-tailed t-test. *p < 0.05. c Volcano plot showing a total of 1483 differentially expressed genes in primary acinar cells isolated from Prdm3ΔAcinar mice, relative to control Ptf1aCreER. Individual genes are labeled and circled in black. d Volcano plot showing differentially expressed genes belonging to inflammatory responses and NF-κB signaling are highlighted in red and blue, respectively. Gene Set Enrichment Analysis (GSEA) of differentially expressed genes (Ptf1aCreER vs. Prdm3ΔAcinar) identified enrichment of immune responses and regulation of NF-κB signaling. Normalized enrichment score (NES) and p-values are shown. e Functional annotation on 1073 upregulated genes in primary acinar cells isolated from Prdm3ΔAcinar mice, relative to control. Significant KEGG and Hallmark terms, p-values and ranks are shown.
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References

    1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. CA Cancer J. Clin. 2019;69:7–34. doi: 10.3322/caac.21551. - DOI - PubMed
    1. Murtaugh LC, Keefe MD. Regeneration and repair of the exocrine pancreas. Annu. Rev. Physiol. 2015;77:229–249. doi: 10.1146/annurev-physiol-021014-071727. - DOI - PMC - PubMed
    1. Storz P. Acinar cell plasticity and development of pancreatic ductal adenocarcinoma. Nat. Rev. Gastroenterol. Hepatol. 2017;14:296–304. doi: 10.1038/nrgastro.2017.12. - DOI - PMC - PubMed
    1. Morris JPt, Cano DA, Sekine S, Wang SC, Hebrok M. Beta-catenin blocks Kras-dependent reprogramming of acini into pancreatic cancer precursor lesions in mice. J. Clin. Invest. 2010;120:508–520. doi: 10.1172/JCI40045. - DOI - PMC - PubMed
    1. Kopp JL, et al. Identification of Sox9-dependent acinar-to-ductal reprogramming as the principal mechanism for initiation of pancreatic ductal adenocarcinoma. Cancer Cell. 2012;22:737–750. doi: 10.1016/j.ccr.2012.10.025. - DOI - PMC - PubMed

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