Pancreatic ductal adenocarcinoma (PDAC) has been widely considered as one of the most lethal malignancies with a 5-year overall survival (OS) rate of only 11%.1 Lots of efforts have been devoted into its early diagnosis, treatment strategies, and pathogenesis, etc. PDAC is characterized as excessive desmoplastic tumor with abundant immune cells infiltration, mainly including macrophages, T cells, immature myeloid cells, and neutrophils. Neutrophils are the most abundant white blood cells in blood and have been regarded as a homogeneous group in the past, which belongs to innate immune system and participates in defensing against pathogens. Recently, accumulating evidences determined that neutrophils had an important role in PDAC and accounted for a substantial proportion of tumor-infiltrating immune and inflammatory cells. Jiang et al.2 found that increased neutrophil infiltration was discovered as a central and prominent affected feature, which occurred in the liver, lung, and stomach at the PanIN stage. Importantly, serum leukotriene B4 (LTB4), derived from neutrophils, was validated for the early detection of PDAC. Considering its prognostic role, Ino et al.3 described relationships between prognosis and infiltrating immune cells in a cohort enrolled 212 PDAC patients who received radical surgical resection and found that tumor-infiltrating neutrophils were positively correlated with macrophages and regulatory T cells infiltrations and closely associated with shorter OS and disease-free survival (FS). This indicated that high tumor-infiltrating neutrophils always present boost immunosuppressive microenvironment and contribute to poor survival. Apart from tumor-infiltrating neutrophils, higher circulating neutrophils have also been found a negative association with outcomes. In our previous report, we noted that circulating neutrophil counts, whether 3 days within preoperation or 1 day after operation, were associated with shorter recurrence-FS (RFS) but not with OS, which may hinted that neutrophils exerted a pro-tumor effect in PDAC.4 Tumor-infiltrating neutrophils were often referred as tumor-associated neutrophils (TANs) in most studies. Wang et al.5 identified a pro-tumor subcluster of neutrophils in PDAC and uncovered the pro-tumor mechanisms of TANs in PDAC microenvironment and revealed the association between high glycolytic activity and pro-tumor functions in TANs. Similar to M1/M2 nomenclature of macrophages, recent studies also suggested that TANs could be divided into N1 and N2 categories based on their distinctive phenotypes: antitumorigenic N1 neutrophils and pro-tumorigenic N2 neutrophils. According to current in vivo/vitro studies, the N1/N2 neutrophil polarization may be depended on the special cytokine milieu, mainly including interferon-β (IFN-β) and TGF-β. However, little is known about the prognostic value of tumor-associated N1/N2 neutrophils in PDAC. We preliminarily collected the PDAC tissues after radical surgery from January 2012 to December 2015 in our institute and stained for tumor-associated N1 and N2 neutrophils as refered.6, 7 The typical cell markers of tumor-associated N1 neutrophils were MPO+CD11b+CD206−, and those of tumor-associated N2 neutrophils were MPO+CD11b+CD206+ by three-color immunofluorescence (IF) staining (Figure 1A). Studies have reported that N1 neutrophils may possess powerful antitumor properties through antibody-dependent or direct cytotoxicity8, ROS-mediated coupling9, etc. On the contrary, N2 neutrophils contributed to tumor angiogenesis by secreting vascular endothelial growth factor and matrix metallopeptidase 9, etc. and suppressing CTLs function by arginase.10 In this study, a total of 77 PDAC patients were preliminarily enrolled. Their median age was 65.0 [interquartile range (IQR) 59.5–70.0] years old, and 63.6% of patients (49 of 77) were male. Approximately 25% and 40% of PDAC patients have reached advanced T stages (T3 and T4) and aggressive N stages (N1 and N2) when surgery, respectively. Other clinicopathological characteristics are displayed in Table S1. The median number of tumor-associated N1 and N2 neutrophils were 10.7 (IQR 7.2–25.2) and 21.0 (IQR 13.7–29.0), respectively. The unpaired analysis indicated that the mean number of tumor-associated N2 neutrophils was 23.0 higher than that of tumor-associated N1 neutrophils (16.7, p = 0.002, Figure 1B). Additionally, the discrepancy between the tumor-associated N1 and N2 neutrophils were also observed in the paired analysis (p = 0.011, Figure 1C). According to the optimal cutoffs of tumor-associated N1 and N2 neutrophils, 22 and 18 cases were respectively classified into the high N1 and N2 neutrophil infiltration group. As shown in Table S1, the lower tumor-associated N1 neutrophil infiltration was significantly associated with easier lymph node metastasis (p = 0.012) and higher TNM stage (p = 0.024), but not with other factors. The higher tumor-associated N2 neutrophil infiltration was significantly correlated with distal location (p = 0.039) and easier lymph node metastasis (p = 0.039), similar to the previous study that more aggressive PDAC preferentially recruited more neutrophils. The median OS and RFS of this cohort were 20 and 15 months, respectively. There were 81.6%, 34.1%, and 26.5% OS rates at 1, 3, and 5 years, respectively, while the 1-, 3-, and 5-year RFS rates were 57.9%, 24.9%, and 19.6%, respectively. The Kaplan–Meier survival analyses showed a significant difference in median OS and RFS between the high tumor-associated N1 neutrophil group and the lower group [OS: not reached versus 18 months, Log-rank p < 0.001, HR = 0.153, 95% confidence interval (CI) 0.089–0.262, Figure 1D; RFS: 84 versus 12 months, Log-rank p < 0.001, HR = 0.209, 95% CI 0.125–0.348, Figure 1E]. Accordingly, the median OS (12 versus 32 months, Log-rank p < 0.001, HR = 5.352, 95% CI 2.066–13.870, Figure 1F) and RFS (8.5 versus 22 months, Log-rank p < 0.001, HR = 3.852, 95% CI 1.660–8.941, Figure 1G) of the more markable tumor-associated N2 neutrophil infiltration group were both statistically shorter. Meanwhile, the univariate and multivariate Cox analyses together with conventional clinicopathological variables demonstrated that both tumor-associated N1 and N2 neutrophils were independent prognostic factors for OS (Table S2) and RFS (Table S3) with the opposite HR values. Accumulative evidences showed that neutrophils had complex interaction with tumor microenvironment and played an important role in PDAC progression. The predictive values of neutrophil relevant biomarkers have been found in PDAC patients and regarded as potential therapeutic target. High density of total TANs infiltration always indicated poor outcome in PDAC, and current strategies of anti-TANs mainly focused on depleting TANs entirely or blocking chemokines that functioned as TANs recruitment. Series of preclinical studies used Ly6G or CXCR2 to block TANs and had encouraging outcomes in different mouse models. However, these therapeutic strategies are fraught with difficulties for further clinical trials because of the vital function in defense pathogens of neutrophils. It is worth to consider that TANs have the opposite phenotype between N1 and N2. Our study successfully found that both tumor-associated N1 and N2 neutrophils displayed significant and opposite prognostic values within PDAC. The N1/N2 polarization may depend on different stimuli, including TGF-β induces N2 phenotype whereas IFN-β signaling polarizes neutrophils to N1 phenotype, and the causes of opposite effect in N1 and N2 neutrophil may include the secretions of different cytokines that orchestrate immune cells recruitment, different capability of arginase and proteases synthesis, and direct or indirect cytotoxicity, etc. The aforementioned results hinted that precisely eliminating N2 by specific markers or modified/polarized TANs into N1 would be a more efficacious strategy with less toxicity. In summary, the exploration of neutrophil polarization and its correlation to clinical features in PDAC would significantly enhance our understanding of its pathophysiology and enable us to develop better treatment options. Study conception and design: NP and HY. Acquisition of data: HY, SG, QC, SL, and NP. Analysis and interpretation: HY, SG, QC, SL, SS, YJ, WL, JY, WW, and NP. Drafting of manuscript: NP and HY. Critical revision: HY, SG, QC, SL, SS, YJ, WL, JY, WW, and NP. Statistical analysis: HY, SG, QC, and NP. Study supervision: NP and WW. Read and approved the final manuscript: HY, SG, QC, SL, SS, YJ, WL, JY, WW, and NP. The research was supported by grants from the National Natural Science Foundation of China (grant number: 82103409), China Postdoctoral Science Foundation (grant number: 2021M690037), Shanghai Sailing Program (grant numbers: 21YF1407100 and 21YF1443500), Health Science Popularization Project of Pancreatic Cancer of Zhongshan Hospital (grant number: ZX2021-002), Youth Fund of Zhongshan Hospital Fudan University (grant number: LCBSHZX010), and Shanghai ShenKang Hospital Development Centre Project (grant number: SHDC2020CR2017B). The authors declare no conflict of interest. The study protocol was reviewed and approved by the Ethics Committee of Zhongshan Hospital, Fudan University. Data included in this study are available upon request by contact with the corresponding author. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. 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Abstract BACKGROUND : Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies. Its larger mass size is widely acknowledged to be associated with increased lymph node (LN) metastatic potential. However, the quantitative relationships between tumor size and LN metastasis or survival remain unclear. Thus, this study aims to quantitatively identify the objective relationship between tumor size and prevalence of LN metastases across primary tumor size spectrums. METHODS : 9,958 resected PDAC patients without distant metastasis were retrieved from the Surveillance, Epidemiology, and End Results (SEER) database. The prevalence of LN metastases, LN ratio (LNR) and N2/N1 ratio were assessed amongst different tumor sizes, and the relationships were depicted by matched curves. RESULTS : In the enrolled cohort, age, tumor site, grade, American Joint Committee on Cancer (AJCC) 8th node staging, tumor size, chemotherapy and radiotherapy were identified as significant independent predictors for overall survival (OS) and cancer-specific survival (CSS). For tumors within 1-40 mm in size, the prevalence of node positive disease is closely modelled using a logarithmic formula [0.249×ln (size) + 0.452] × 100%, and then fluctuated between 70.0% and 80.0% when beyond 40 mm. The mean LNR increased in a stepwise manner as tumor size increased from 1-5 mm (LNR=0.024) to 41-45 mm (LNR=0.177); then, beyond 45 mm, it plateaued near 0.170. N2/N1 ratio gradually increased along with tumor size from 1-5 mm (N2/N1=0.286) to 41-45 mm (N2/N1=1.016), and when tumor size reached to 41-45 mm or more, the ratio stabilized around 1.000. CONCLUSION : Regional LN involvement demonstrated a logarithmic growth with increasing tumor sizes in resectable PDAC patients. The probability of metastasis in each regional LN for resectable PDAC patients with tumors greater than 40 mm in size was near 17.0% and their overall prevalence of LN metastasis was 70%-80%. Among which, 50% of patients had an N2 stage.
Abstract Pancreatic cancer (PC), lacking biomarkers and effective therapeutics, remains highly lethal. Data regarding the correlations of PC risk and the individual plasma proteome known for minimally cancer biomarkers, are scarce. Here, we measure 1,345 human plasma proteins via Proteome-Wide Association Studies, presenting 78 proteins are prominently related to PC risk, including 4 proteins (ROR1, FN1, APOA5, ABO) exhibit the strongest causal association identified via Mendelian Randomization and Colocalization. Our two independent cohorts further demonstrate FN1 and ABO are highly expressed in blood or tumors from patients with PC compared to specimens from healthy individuals or para-tumors. Moreover, patients with higher levels of FN1 and ABO in their blood or tumors have worse median survival than those with lower levels. Multiple drugs targeting FN1 are currently available or undergoing clinical testing, making FN1 a promisingly repurposed therapeutic target in addition to severing as a circulating prognostic indicator for PC.
Abstract Light stimulation can realise the remote control of the deformation of the specific position of 4D printing structure. Shape-memory polymer–carbon nanotube (CNT) composite materials, with outstanding near-infrared photothermal conversion rate and shape-memory ability, is one type of the most popular light responsive smart materials. However, current studies focused on the photothermal effect and shape-memory applications of light-responsive shape-memory polymer composite (SMPC) sheet structures, and there is no research on the photothermal effect in the depth direction of light-responsive SMPC three-dimensional structures. Here, we prepared a UV curable, mechanically robust, and highly deformable shape-memory polymer (IBBA) as the matrix of light responsive SMPC. CNTs were added as photothermal conversion materials. We explore the photothermal effect of near-infrared laser on the surface and depth of IBBA–CNT composites cube. Shape-memory experiments show that different folded shapes can be obtained by selective near-infrared laser programming. Selective near-infrared laser programming three-dimensional movable type plate shows a programming application in depth direction of three-dimensional light-responsive intelligent structure. This research extends the application of near-infrared laser in 4D printing to the depth direction of intelligent structures, which will bring more complex and interesting 4D printing structures in the future.
Radiofrequency ablation (RFA) is widely used in palliative therapy of malignant cancers. Several studies have shown its applicability and safety for locally advanced pancreatic cancer (LAPC). The objective of this study was to modify the current regimen to improve its therapeutic effect.Immune cell subtypes and related cytokines were quantified to uncover the immune pattern changes post-RFA treatment. Then, high-throughput proteome analysis was performed to identify differentially expressed proteins associated with RFA, which were further validated in in vitro and in vivo experiments. Finally, a combined therapy was tested in a murine model to observe its therapeutic effect.In preclinical murine models of RFA treatment, no significant therapeutic benefit was observed following RFA treatment. However, the proportion of tumor-infiltrating CD8+ T cells was significantly increased, whereas that of regulatory T cells (Tregs) was decreased post-RFA treatment, which indicated a beneficial anti-tumor environment. To identify the mechanism, high-throughput mass spectrum was obtained that identified heat shock protein 70 (HSP70) as the top differentially expressed protein. HSP70 expression in residual cancer cells was significantly increased post-RFA treatment, which notably promoted pancreatic cancer growth. Elevated HSP70 promoted cell proliferation by activating AKT-mTOR signaling. Finally, RFA treatment combined with an mTOR inhibitor exerted a synergetic repressive effect on tumor growth in the preclinical murine cancer model.RFA treatment in combination with mTOR signaling blockade can not only promote tumor immune response, but also restrain residual cancer cell proliferation. Such a combination may be a promising and effective therapeutic strategy for LAPC patients.
Background: The prognosis of pancreatic carcinoma (PC) remains poor and the American Joint Committee on Cancer (AJCC) 8th staging system for survival prediction in PC patients after curative resection is still limited. Thus, the aim of this study is to refine a valuable prognostic model and novel staging system for PC with curative resection. Methods: The data of 3,458 patients used in this study were retrieved from the Surveillance, Epidemiology, and End Results database registry of National Cancer Institute. The prognostic value of lymph node ratio (LNR) was analyzed in the primary cohort and prognostic nomogram based on the LNR was established to create a novel staging system. Then, analyses were conducted to evaluate the application of the formulated nomogram staging system and the AJCC 8th staging system. The predictive performance of model was further validated in the internal validation cohort. Results: Significant positive correlations were found between LNR and all factors except for surgical procedures. The results of univariate and multivariate analyses showed that LNR was identified as an independent prognostic indicator for overall survival (OS) in both primary and validation cohorts (all P < 0.001). A prognostic nomogram based on the LNR was formulated to obtain superior discriminatory abilities. Compared with the AJCC 8th staging system, the formulated nomogram staging system showed higher hazard ratios of stage II, III, and IV disease (reference to stage I disease) that were 1.637, 2.300, and 3.521, respectively, by univariate analyses in the primary cohort and the distinction between stage I, II, and III disease at the beginning or end of the survival curves was more apparent. All these results were further verified in the validation cohort. Conclusion: LNR can be considered as a useful independent prognostic indicator for PC patients after curative resection regardless of the surgical procedures. Compared with the AJCC 8th staging system, the formulated nomogram showed superior predictive accuracy for OS and its novel staging system revealed better risk stratification. Keywords: pancreatic head carcinoma, lymph node ratio, nomogram, prognosis, decision curve analysis, AJCC
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