YAP is required for prostate development, regeneration, and prostate stem cell function.

Prostate development and regeneration depend on prostate stem cell function, the delicate balance of stem cell self-renewal and differentiation. However, mechanisms modulating prostate stem cell function remain poorly identified. Here, we explored the roles of Yes-associated protein 1 (YAP) in prostate stem cells, prostate development and regeneration. Using YAPfl/fl, CD133-CreER mice, we found that stem cell-specific YAP-deficient mice had compromised branching morphogenesis and epithelial differentiation, resulting in damaged prostate development. YAP inhibition also significantly affected the regeneration process of mice prostate, leading to impaired regenerated prostate. Furthermore, YAP ablation in prostate stem cells significantly reduced its self-renewal activity in vitro, and attenuated prostate regeneration of prostate grafts in vivo. Further analysis revealed a decrease in Notch and Hedgehog pathways expression in YAP inhibition cells, and treatment with exogenous Shh partially restored the self-renewal ability of prostate sphere cells. Taken together, our results revealed the roles of YAP in prostate stem cell function and prostate development and regeneration through regulation of the Notch and Hedgehog signaling pathways.

Amoeba-inspired magnetic microgel assembly assisted by engineered dextran-binding protein for vaccination against life-threatening systemic infection.

Vaccination is critical for population protection from pathogenic infections. However, its efficiency is frequently compromised by a failure of antigen retention and presentation. Herein, we designed a dextran-binding protein DexBP, which is composed of the carbohydrate-binding domains of Trichoderma reesei cellobiohydrolases Cel6A and Cel7A, together with the sequence of the fluorescent protein mCherry. DexBP was further prepared by engineered Escherichia coli cells and grafted to magnetic nanoparticles. The magnetic nanoparticles were integrated with a dextran/poly(vinyl alcohol) framework and a reactive oxygen species-responsive linker, obtaining magnetic polymeric microgels for carrying pathogen antigen. Similar to amoeba aggregation, the microgels self-assembled to form aggregates and further induced dendritic cell aggregation. This step-by-step assembly retained antigens at lymph nodes, promoted antigen presentation, stimulated humoral immunity, and protected the mice from life-threatening systemic infections. This study developed a magnetic microgel-assembling platform for dynamically regulating immune response during protection of the body from dangerous infections. Electronic Supplementary Material: Supplementary material (AFM image and zeta potential of MG; TEM, FT-IR, DLS, and zeta potential of MNP-DexBP; zeta potential of MG+CaAg and MG+MNP-DexBP+CaAg; antigen release profile of MG+CaAg and MG+MNP-DexBP+CaAg; aggregation and dispersion of dendritic cells induced by MG+MNP-DexBP+CaAg; uptake of FITC-labeled CaAg (fCaAg) and intracellular distribution of fCaAg in the dendritic cells; antigen retention and dendritic cell activation in lymph nodes; and serum anti-CaAg antibody levels on day 3 after C. albicans infection in the mice pre-immunized by PBS (control), CaAg, MG+CaAg, and MG+MNP-DexBP+CaAg) is available in the online version of this article at 10.1007/s12274-022-4809-1.

Economical and Efficient Protocol for Isolating and Culturing Bone Marrow-derived Dendritic Cells from Mice.

The demand for dendritic cells (DCs) is gradually increasing as immunology research advances. However, DCs are rare in all tissues. The traditional method for isolating DCs primarily involves inducing bone marrow (BM) differentiation into DCs by injecting large doses (>10 ng/mL) of granulocyte-macrophage colony-stimulating factor/interleukin-4 (GM-CSF/IL-4), making the procedure complex and expensive. In this protocol, using all BM cells cultured in 10 ng/mL GM-CSF/IL-4 medium, after 3-4 half-culture exchanges, up to 2.7 x 107 CD11c+ cells (DCs) per mouse (two femurs) were harvested with a purity of 80%-95%. After 10 days in culture, the expression of CD11c, CD80, and MHC II increased, whereas the number of cells decreased. The number of cells peaked after 7 days of culture. Moreover, this method only took 10 min to harvest all bone marrow cells, and a high number of DCs were obtained after 1 week of culture.

Assessing the Potential Prognostic and Immunological Role of TK1 in Prostate Cancer.

Background: It has been reported that thymidine kinase 1 (TK1) was up-regulated in multiple malignancies and participated in the regulation of tumor malignant behavior. However, its specific role in prostate cancer (PCa) remains unclear. Methods: TK1 expression in PCa patients and cell lines was identified via crossover analysis of the public datasets. A series of in vitro experiments and in vivo models was applied to investigate the function of TK1 in PCa. Functional enrichment analyses were further conducted to explore the underlying mechanism. Additionally, TISIDB was applied to explore the correlation between TK1 expression and tumor-infiltrating lymphocytes, immune subtypes, and immune regulatory factors. Results: TK1 expression was significantly up-regulated in PCa patients and cell lines. TK1 ablation inhibited tumor cell proliferation and migration potential, and in vivo experiments showed that TK1 inactivation can significantly restrain tumor growth. Functional enrichment analysis revealed TK1-related hub genes (AURKB, CCNB2, CDC20, CDCA5, CDK1, CENPA, CENPM, KIF2C, NDC80, NUF2, PLK1, SKA1, SPC25, ZWINT), and found that TK1 was closely involved in the regulation of cell cycle. Moreover, elevated mRNA expression of TK1 was related with higher Gleason score, higher clinical stage, higher pathological stage, higher lymph node stage, shorter overall survival, and DFS in PCa patients. Particularly, TK1 represented attenuated expression in C3 PCa and was related with infiltration of CD4+, CD8+ T cells, and dendritic cells as well as immunomodulator expression. Conclusion: Our study indicates that TK1 is a prognostic predictor correlated with poor outcomes of PCa patients, and for the first time represented that TK1 can promote the progression of PCa. Therefore, TK1 may be a potential diagnostic and prognostic biomarker, as well as a therapeutic target for PCa.

Fusion Protein Vaccine Based on Ag85B and STEAP1 Induces a Protective Immune Response against Prostate Cancer.

(1) Background: There are currently limited treatments for castration-resistant prostate cancer. Immunotherapy involving Sipuleucel-T has increasingly drawn attention for prostate cancer management. BCG plays a vital role in treating bladder cancer, mainly by inducing immune activation, but is rarely used for prostate cancer. (2) Methods: The TCGA database, PCR, and Western blotting were used to analyze the expression of STEAP1 in mouse and human tissues. Then, we constructed a fusion protein vaccine with Mycobacterium tuberculosis Ag85B and three repeated octapeptide epitopes of a six-transmembrane epithelial antigen of the prostate 1 (STEAP1186-193), Ag85B-3×STEAP1186-193. The uptake of the fusion protein vaccine by DCs was evaluated by confocal microscopy, and DC markers were detected using flow cytometry after incubation with the fusion protein. The immune response against prostate cancer was evaluated by the LDH assay and xenografts in vitro and in vivo. Then, the tumor microenvironment was determined using IHC and ELISA. In addition, the epitope was mutated using CRISPR-Cas9 to illustrate that the fusion protein elicited immunization against STEAP1. (3) Results: The TCGA database analysis, PCR, and Western blotting showed that STEAP1 was highly expressed in human and murine prostate cancer. After the uptake of the purified fusion protein vaccine by DCs, CD11c, CD80, CD86, and MHC II were upregulated and triggered a cytotoxic T lymphocyte (CTL) response against TRAMP-C1 and RM1 cells in vitro. Furthermore, the fusion protein vaccine inhibited tumor growth and improved the tumor microenvironment in vivo, with more CD3+ cells and fewer FOXP3+ cells in the tumor. Serum IFN-γ and IL-2 were significantly higher than in the control group, while IL-4 expression was lower, indicating that the fusion protein vaccine activated Th1 immunity. The immune response against prostate cancer was greatly suppressed when the antigen targets were knocked out using CRISPR-Cas9. (4) Conclusion: In summary, our results provide the first evidence that a vaccine based on a fusion protein consisting of Ag85B and a prostate cancer octapeptide epitope with complete Freund's adjuvant (CFA), triggers a robust immune response and inhibits tumor growth in murine prostate cancer.

Correction: Prognostic and clinicopathological value of p53 expression in renal cell carcinoma: a meta-analysis.

[This corrects the article DOI: 10.18632/oncotarget.21971.].

Neutrophil-lymphocyte ratio is a predictor of prognosis in patients with castration-resistant prostate cancer: a meta-analysis.

BACKGROUND: The prognostic value of neutrophil-lymphocyte ratio (NLR) in patients with castration-resistant prostate cancer (CRPC) had been investigated in previous studies; however, the results remain inconsistent. This study was aimed to investigate the prognostic value of NLR in CRPC patients. MATERIALS AND METHODS: Literature was identified from PubMed, Embase, Web of Science, and Cochrane, which investigated the relationship between pretreatment NLR and prognosis in CRPC patients. HRs for overall survival (OS) and progression-free survival (PFS) were extracted from eligible studies. Heterogeneity was assessed using the I2 value. The fixed-effects model was used if there was no evidence of heterogeneity; otherwise, the random-effects model was used. Publication bias was evaluated using Begg's funnel plot test. RESULTS: A total of 5,705 patients from 16 studies were included in this analysis. The pooled results showed that an elevated NLR predict poor OS (pooled HR = 1.52, 95% CI: 1.41-1.63, P<0.001) and PFS (pooled HR = 1.50, 95% CI: 1.21-1.85, P<0.001) in patients with CRPC. Subgroup analysis revealed that an elevated NLR significantly predicted poor OS in Asian studies group (HR = 2.43, 95% CI: 1.47-4.01, P=0.001). The elevated NLR also significantly predicted poor PFS in Asian studies group (HR = 1.99, 95% CI: 1.30-3.06, P=0.002). CONCLUSION: This study suggests that an elevated NLR predict poor prognosis in patients with CRPC.

Prognostic and clinicopathological value of PBRM1 expression in renal cell carcinoma.

BACKGROUND: The prognostic value of PBRM1 expression in renal cell carcinoma (RCC) had been investigated in previous studies; however, the results remain inconclusive. We investigated the prognostic value and clinicopathological significance of PBRM1 protein expression in RCC. METHODS: PubMed, Embase, Web of Science, and Cochrane database were searched for studies investigating the relationships between PBRM1 expression and outcomes in RCC. Hazard ratios (HRs) for survival outcomes and odds ratios (ORs) for clinical parameters were extracted from eligible studies. Heterogeneity was assessed using the I2 value. The fixed-effects model was used if there was no evidence of heterogeneity; otherwise, the random-effects model was used. Publication bias was evaluated using Begg's funnel plots and Egger's regression test. RESULTS: A total of 2942 patients from 7 studies were included in the meta-analysis. The results showed that decreased expression of PBRM1 is associated with poor overall survival (OS) (HR = 2.11, 95% CI: 1.52-2.96), cancer-specific survival (CSS) (HR = 1.32, 95% CI: 1.10-1.58), and progression-free survival/ recurrence-free survival (PFS/RFS) (HR = 1.57, 95%CI: 1.34-1.85) in RCC. In addition, PBRM1 positive expression was significantly associated with earlier TNM stage (III/IV vs. I/II, OR = 0.53, 95% CI: 0.30-0.94), primary tumor stage (pT3/4 vs. pT1/2, OR = 0.32, 95% CI: 0.20-0.52), and Fuhrman grade (3/4 vs. 1/2, OR = 0.69, 95% CI: 0.46-1.02), but not related to Necrosis or Sex. CONCLUSIONS: Decreased expression of PBRM1 is correlated with poor prognosis and advanced clinicopathological features in patients with RCC.

Platelet-lymphocyte ratio acts as an independent predictor of prognosis in patients with renal cell carcinoma.

BACKGROUND: The prognostic value of plated-lymphocyte ratio (PLR) in multiple malignancies had been investigated in previous studies; however, its prognostic value in renal cell carcinoma (RCC) remains controversial. This study was performed to assess the prognostic value of preoperative PLR in RCC patients. METHODS: Literature was searched from PubMed, Embase, Web of Science and Cochrane database, which evaluated the relationships between preoperative PLR and prognosis in RCC patients. Hazard ratios (HRs) for overall survival (OS) and progression-free survival (PFS) were extracted from eligible studies. Heterogeneity was assessed using the I2 value. The fixed-effects model was used if there was no evidence of heterogeneity; otherwise, the random-effects model was used. Publication bias was evaluated using Begg's funnel plots and Egger's regression test. RESULTS: A total of 1528 patients from seven studies were included in the analysis. The pooled analysis showed that an elevated PLR was an effective prognostic marker of both OS (pooled HR = 2.10, 95%CI: 1.38-3.19, p = 0.001) and PFS (pooled HR = 3.45, 95%CI: 1.61-7.40, p = 0.001). Subgroup analysis revealed that a high PLR significantly predicted worse OS and PFS in Asian studies (OS, pooled HR = 2.72, 95%CI: 1.06-7.03, p = 0.038; PFS, pooled HR = 6.0, 95%CI: 3.12-11.54, p < 0.001), in metastatic RCC patients receiving mixed therapies (OS, pooled HR = 3.69, 95%CI: 1.93-11.42, p = 0.023; PFS, pooled HR = 6.05, 95%CI: 1.34-27.37, p = 0.019) and targeted therapy (OS, pooled HR = 1.59, 95%CI: 0.97-2.62, p = 0.067), in sample size >100 (OS, pooled HR = 1.83, 95%CI: 1.49-2.25, p < 0.001; PFS pooled HR = 6.05, 95%CI: 1.34-27.37, p < 0.019), and in cut-off value of PLR ≤ 195 (OS, pooled HR = 3.65, 95%CI: 1.06-12.60, p = 0.04; PFS pooled HR 4.46, 95%CI: 1.68-11.87, p = 0.003). CONCLUSIONS: This study suggests that a high preoperative PLR is correlated with poor prognosis in RCC patients.

Prognostic and clinicopathological significance of PD-L1 in patients with renal cell carcinoma: a meta-analysis based on 1863 individuals.

The prognostic significance of PD-L1 in renal cell carcinoma (RCC) had been investigated in previous studies; however, the results remain controversial. The primary aim of this meta-analysis was to investigate the prognostic and clinicopathological significance of the PD-L1 expression in patients with RCC. Relevant literature was identified form PubMed, Embase, Web of Science and Cochrane library, which compared the prognostic significance between PD-L1 expression and RCC. Hazard ratios (HRs) for survival outcomes and odds ratios (ORs) for clinical parameters associated with PD-L1 were extracted from eligible studies. Heterogeneity was assessed using the I2 value. The fixed-effects model was used if there was no evidence of heterogeneity; otherwise, the random-effects model was used. Publication bias was evaluated using Begg's funnel plots and Egger's regression test. A total of 1863 patients from ten eligible studies were analyzed. The results showed that PD-L1 expression is associated with poor overall survival in clear cell RCC (ccRCC) (HR = 2.76, 95%CI: 2.25-3.38, I2 = 14.4%, P < 0.001) and non-clear cell RCC (non-ccRCC) (HR = 2.77, 95%CI: 1.62-4.72, I2 = 28.8%, P < 0.001). In addition, PD-L1 expression was found to be significantly associated with primary tumor stage (OR = 1.76, 95%CI: 1.39-2.23; I2 = 56.3%), regional lymph node involvement (OR = 2.10, 95%CI: 1.48-2.98; I2 = 14.9%), distant metastases (OR = 2.69, 95%CI: 2.05-3.54; I2 = 0.0%), nuclear grade (OR = 1.72, 95%CI: 1.32-2.23; I2 = 79.4%) and histologic tumor necrosis (OR = 2.25, 95%CI: 1.59-3.18; I2 = 66.1%) in patients with RCC. The outcome stability was confirmed by sensitivity analysis. Both the Begg's funnel plot test (P = 0.276) and the Egger's (P = 0.388) verified that there was no publication bias within the included studies. This study suggests that PD-L1 expression is correlated with poor prognosis and advanced clinicopathological features in RCC patients.

Prognostic and clinicopathological value of Ki-67/MIB-1 expression in renal cell carcinoma: a meta-analysis based on 4579 individuals.

BACKGROUND: Previous studies have investigated the prognostic significance of Ki-67/MIB-1 expression in renal cell carcinoma (RCC), however, the reports are controversial and inconsistent. This study aimed to investigate Ki-67/MIB-1 expression in RCC and its correlation with prognosis and clinicopathological features. METHODS: We searched relevant studies that reported associations between Ki-67/MIB-1 expression and prognosis in RCC from PubMed, Embase, Web of Science, and Cochrane Library studies published until April 14, 2017. Hazard ratios (HRs) and 95% confidence intervals (CIs) were extracted from eligible studies. Fixed and random effects models were used to calculate pooled HRs and 95% CIs according to heterogeneity. RESULTS: A total of 4579 participants from 23 eligible studies were included in this analysis. The results showed that Ki-67/MIB-1 expression was associated with poor overall survival (HR=2.06, 95% CI: 1.64-2.57) and cancer specific survival (HR=2.01, 95% CI: 1.66-2.44). In addition, Ki-67/MIB-1 expression was also correlated with TNM stage (III/IV vs I/II: OR=1.92, 95% CI: 1.61-2.28), pathological T stage (pT3/pT4 vs pT1/pT2: OR=1.56, 95% CI: 1.21-2.02), distant metastasis (M1 vs M0: OR=1.81, 95% CI: 1.34-2.43), and Fuhrman grade (III/IV vs I/II: OR=1.94, 95% CI: 1.21-3.10). CONCLUSION: Our study demonstrates that the presence of high Ki-67/MIB-1 expression and advanced clinicopathological features were correlated with poor prognosis in RCC patients.

Prognostic and clinicopathological value of p53 expression in renal cell carcinoma: a meta-analysis.

BACKGROUND: The prognostic value of p53 expression in renal cell carcinoma (RCC) had been investigated in previous studies; however, the results remain inconsistent. This study was performed to investigate the prognostic and clinicopathological significance of p53 protein expression in RCC. MATERIALS AND METHODS: Literature was identified from PubMed, Embase, Web of Science, and Cochrane database, which investigated the relationships between p53 expression and outcomes. Hazard ratios (HRs) for survival outcomes and odds ratios (ORs) for clinical parameters associated with p53 were extracted from eligible studies. Heterogeneity was assessed using the I2 value. The fixed-effects model was used if there was no evidence of heterogeneity; otherwise, the random-effects model was used. Publication bias was evaluated using Begg's funnel plots and Egger's regression test. RESULTS: A total of 2,013 patients from 22 studies were included in the meta-analysis. The results showed that p53 positive expression is associated with poor overall survival (OS) (HR = 2.17, 95% confidence [CI]: 1.51-3.13) and cancer-specific survival (CSS) (HR = 1.59, 95% CI: 1.19-2.12) in RCC. In addition, p53 positive expression was closely correlated with TNM stage (III/IV vs. I/II: OR = 2.51, 95% CI: 1.05-6.00), Fuhrman grade (III/IV vs. I/II: OR = 1.80, 95% CI: 1.24-2.63), and distant metastasis (M1 vs. M0: OR = 1.70, 95% CI: 1.16-2.49), but not related to lymph node involvement (N1 vs. N0: OR = 1.32, 95% CI: 0.80-2.18), primary tumor stage (pT3/pT4 vs. pT1/pT2: OR = 1.16, 95% CI: 0.88-1.53), and sex (n = 2, male vs. female, OR = 1.09, 95% CI: 0.70-1.68). CONCLUSIONS: This study suggests that p53 positive expression is correlated with poor prognosis and advanced clinicopathological features in patients with RCC, which indicates that p53 is a potentially effective therapeutic target.

C-reactive protein is a predictor of prognosis in renal cell carcinoma patients receiving tyrosine kinase inhibitors: A meta-analysis.

BACKGROUND: The prognostic value of C-reactive protein (CRP) in metastatic renal cell carcinoma (RCC) patients receiving tyrosine kinase inhibitors (TKIs) has been investigated in previous studies; however, the results remain inconclusive. This study investigated the prognostic value of pretreatment CRP in patients with metastatic RCC treated with TKIs. METHODS: PubMed, Embase, Web of Science, and Cochrane databases were searched for studies investigating the relationships between pretreatment CRP and prognosis in patients with metastatic RCC. Hazard ratios (HRs) for overall survival (OS) and progression-free survival (PFS) were extracted from eligible studies. Heterogeneity was assessed using the I2 value. The fixed-effects model was used if there was no evidence of heterogeneity; otherwise, the random-effects model was used. Publication bias was evaluated using Begg's funnel plots and Egger's regression test. RESULTS: A total of 1199 patients from nine studies were included in the analysis. The results showed that an elevated CRP level was an effective prognostic marker of both OS (pooled HR=2.87, 95% confidence interval [CI]: 2.34-3.54, p<0.001) and PFS (pooled HR=2.39, 95% CI: 1.75-3.26, p<0.001). Subgroup analysis revealed that an elevated CRP level significantly predicted poor OS and PFS in studies conducted in Japan (OS, pooled HR=3.03, 95% CI: 2.29-4.01, p<0.001; PFS, pooled HR=3.6, 95% CI: 1.62-8.0, p=0.002), and in cut-off value of CRP <0.8 (OS, pooled HR=2.93, 95% CI: 2.21-3.88, p<0.001; PFS, pooled HR=2.57, 95% CI: 1.82-3.65, p<0.001). CONCLUSIONS: This study suggests that an elevated CRP level is correlated with poor prognosis in patients with metastatic RCC receiving TKIs treatment.