Dendritic cells (DC) represent a major antigen-presenting cell type in the tumor immune microenvironment (TIME) and play an essential role in cancer immunity. Conventional DC (cDC) and plasmacytoid ...DC (pDC) were defined by the xCell algorithm and a total of 2968 breast cancer patients (TCGA and METABRIC) were analyzed. We found that triple-negative breast cancer (TNBC) had a high fraction of cDC and pDC compared to the other subtypes. In contrast to cDC, high pDC in TNBC was significantly associated with better disease-specific and disease-free survival consistently in both cohorts. High cDC TNBC tumors enriched not only inflammation and immune-related, but also metastasis-related gene sets in Gene Set Enrichment Analysis, whereas high pDC TNBC enriched inflammation and immune -related gene sets including IFN-γ signaling more strongly than cDC. pDC TNBC correlated with CD8+, CD4+ memory, IFN-γ score, and cytolytic activity stronger than cDC TNBC. High pDC TNBC were associated with a high fraction of anti-cancer immune cells and high expression of all the immune check point molecules examined. In conclusion, pDC levels correlated with the infiltration of immune cells and patient survival in TNBC more strongly than cDC; this is the first study suggesting the clinical relevance of pDC infiltration in TNBC.
Annexin A1 (ANXA1) is a phospholipid-linked protein involved in inflammation, immune response, and mast cell reactivity. Recently, we reported that ANXA1 is associated with aggressive features of ...triple-negative breast cancer (TNBC); however, its clinical relevance remains controversial. We hypothesized that human TNBC with high expression of ANXA1 mRNA is associated with pro-cancerous immune cell infiltration, including mast cells, and with an aggressive phenotype. Clinical and RNA-seq data were obtained from The Cancer Genome Atlas (TCGA,
= 1079) and Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) (
= 1904). TNBC patients had significantly higher levels of ANXA1 expression compared to the other subtypes in both TCGA and METABRIC cohorts (
< 0.001). ANXA1 protein expression was assessed by immunohistochemistry in Japanese TNBC patient cohort (
= 48), where 17 cases (35.4%) had positive ANXA1 staining, and their overall survival was significantly shorter compared with negative staining group (
= 0.008). The CIBERSORT algorithm was used to calculate immune cell infiltrations. ANXA1 high tumors were associated with activated mast cells and M2 macrophages (
> 0.01), but did not show any association with tumor heterogeneity nor cytolytic activity. High expression of ANXA1 group enriched inflammation, epithelial-to-mesenchymal transition (EMT), and angiogenesis-related genes in a gene set enrichment assay in both cohorts. To our knowledge, this is the first study to demonstrate that ANXA1 is associated with infiltration of mast cells and inflammation that is associated with the aggressive phenotype of TNBC, such as EMT and angiogenesis.
Purpose
Although the DNA repair mechanism is important in preventing carcinogenesis, its activation in established cancer cells may support their proliferation and aggravate cancer progression. RAD51 ...cooperates with BRCA2 and is essential in the homologous recombination of DNA repair. To this end, we hypothesized that RAD51 gene expression is associated with cancer cell proliferation and poor prognosis of breast cancer (BC) patients.
Methods
A total of 8515 primary BC patients with transcriptome and clinical data from 17 independent cohorts were analyzed. The median value was used to divide each cohort into high and low RAD51 expression groups.
Results
High RAD51 expression enriched the DNA repair gene set and was correlated with DNA repair-related genes. Nottingham histological grade, Ki67 expression and cell proliferation-related gene sets (E2F Targets, G2M Checkpoint and Myc Targets) were all significantly associated with the high RAD51 BC group. RAD51 expression was positively correlated with Homologous Recombination Deficiency, as well as both mutational burden and neoantigens that accompanied a higher infiltration of immune cells. Primary BC with lymph node metastases was associated with high expression of RAD51 in two cohorts. There was no strong correlation between RAD51 expression and drug sensitivity in cell lines, and RAD51 expression was lower after the neoadjuvant chemotherapy compared to before the treatment. High RAD51 BC was associated with poor prognosis consistently in three independent cohorts.
Conclusion
RAD51 gene expression is associated with aggressive cancer biology, cancer cell proliferation, and poor survival in breast cancer.
Cyclosporine A (CsA) is a potent immunosuppressant for treating ulcerative colitis (UC). However, owing to severe systemic side effects, CsA application in UC therapy remains limited. Herein, a ...colon-targeted drug delivery system consisting of CsA crystals (CsAc)-loaded, Eudragit S 100 (ES)-coated alginate microparticles (CsAc-EAMPs) was established to minimize systemic side effects and enhance the therapeutic efficacy of CsA. Homogeneously-sized CsAs (3.1 ± 0.9 μm) were prepared by anti-solvent precipitation, followed by the fabrication of 47.1 ± 6.5 μm-sized CsAc-EAMPs via ionic gelation and ES coating. CsAc-EAMPs exhibited a high drug loading capacity (48 ± 5%) and a CsA encapsulation efficacy of 77 ± 9%. The in vitro drug release study revealed that CsA release from CsAc-EAMPs was suppressed under conditions simulating the stomach and small intestine, resulting in minimized systemic absorption and side effects. Following exposure to the simulated colon conditions, along with ES dissolution and disintegration of alginate microparticles, CsA was released from CsAc-EAMPs, exhibiting a sustained-release profile for up to 24 h after administration. Given the effective colonic delivery of CsA molecules, CsAc-EAMPs conferred enhanced anti-inflammatory activity in mouse model of dextran sulfate sodium (DSS)-induced colitis. These findings suggest that CsAc-EAMPs is a promising drug delivery system for treating UC.
e16002
Background: Among the hallmarks of cancer, epithelial-mesenchymal transition (EMT) that epithelial cancer cells gain the characteristics of mesenchymal cells, is one of the key mechanisms of ...cancer cell metastasis. On the other hand, the clinical relevance of EMT that occurs in gastric cancer patients has not been clearly shown. Methods: To measure the level of EMT in gastric cancer, gene set variation analysis was used to score total of 807 gastric cancer patient samples from two independent large cohorts; TCGA and GSE84437. Results: EMT high gastric cancer was significantly associated with worse overall survival (hazard ratio (HR)=1.74, p=0.011 in the TCGA, and HR=2.01, p<0.001 in the GSE84437), and the association was stronger with EMT signature score compared to expressions of each EMT-related genes, CDH1, CDH2, VIM, or FN1. EMT signature levels were not significantly different among gastric cancer, hepatocellular carcinoma, colorectal cancer, breast cancer, and pancreatic ductal adenocarcinoma, and high level of EMT signature was associated with survival only in gastric cancer. Among the histological types, mucinous and diffuse type had high EMT level compared to tubular, papillary, or signet gastric cancer ( p<0.001). EMT signature level was significantly correlated with tumor depth and AJCC stage (all p<0.001), but not consistently with lymphatic metastasis. Interestingly, the EMT score was independent factor for patient survival including clinicopathological features, not only for overall survival but also disease-specific survival in the TCGA cohort (multivariate; p=0.006 and 0.032, respectively). EMT high gastric cancer was associated with low fraction of Th1 cells and high fraction of dendritic cells and M1 macrophages (all p<0.001 in both cohort), but not with interferon-gamma immune response. EMT high gastric cancer tended to be inversely correlated with cell proliferation-related gene sets, and significantly associated with high infiltration fraction of stromal cells, including fibroblasts, adipocytes (all p<0.001). On the other hand, they significantly enriched multiple pro-cancerous gene sets, such as TGF-β signaling, hypoxia, and angiogenesis, consistent with infiltration of angiogenesis-related stromal cells such as microvascular and lymphatic endothelial cells, and pericytes (all p<0.001). These results were shown consistently in two large cohort; TCGA and GSE84437. Conclusions: EMT signature in a bulk tumor was associated with cancer aggravating pathways such as TGF-β signaling, hypoxia, and angiogenesis, as well as with worse survival in gastric cancer patients.
In this study, we developed pH-triggered surface charge-reversal lipid nanoparticles (LNPs), loaded with budesonide, which could precisely deliver the drug to inflamed colon segments for the ...treatment of ulcerative colitis. Polyethyleneimine (PEI) was used to render LNPs cationic (PEI-LNPs), and Eudragit® S100 (ES) was coated on PEI-LNPs to obtain pH-triggered charge-reversal LNPs (ES-PEI-LNPs). ES coating avoided a burst drug release under acidic conditions mimicking the stomach and early small intestine environments and showed a sustained release in the colon. The surface charge of ES-PEI-LNPs switched from negative to positive under colonic conditions owing to pH-triggered removal of the ES coating. Bioimaging of the mouse gastrointestinal tract and confocal analysis of colon tissues revealed that ES-PEI-LNPs selectively accumulated in an inflamed colon. Furthermore, ES-PEI-LNPs mitigated experimental colitis in mice. These results suggest that the pH-triggered charge-reversal LNPs could be a promising drug carrier for ulcerative colitis therapy and other colon-targeted treatments.
In this study, we developed, through intelligent design, pH-triggered surface charge-reversal lipid nanoparticles, which were shown to precisely deliver a corticosteroid drug to inflamed colon segments in an ulcerative colitis mouse model. The results of the study suggest that the pH-triggered charge-reversal lipid nanoparticles are a promising drug nanocarrier for colon-targeted therapy, including ulcerative colitis treatments. Display omitted
Abstract
BACKGROUND: Currently available breast cancer brain metastasis patient-derived xenografts (PDX) are technically challenging to generate with unstable results. Such PDXs are critical for ...preclinical studies of drug development for treatment of brain metastasis. This is because the biology and microenvironment of metastasis in brain are very different from those of primary tumors or metastases at other organs. Brain metastasis occurs in as many as 30% of patients with advanced breast cancer and is a major cause of mortality. Here, we describe a new method of breast cancer brain metastasis PDXs. We also compare drug responsiveness of orthotopic brain PDXs made with the new method and ectopic PDXs grown in mouse mammary fat pad (MFP).
METHODS: One mm3 fragments of metastatic brain tumors of breast cancer patients were xenografted in brain caudate putamen or MFP of immunodeficient NSG mice using various methods. MRI was used to assess engraftment and tumor growth. Brain and MFP PDXs were examined for responsiveness to epothilone B.
RESULTS: We tested 3 methods for tumor implantation in brain: direct placement of tumor fragment using fine forceps (Forceps method), and injection of gently crushed tumor fragment using beveled 23 G needle with syringe (Needle method) or a pipettor with 10-ul pipette tip of 1 mm bore (Pipette method). PDXs in 8-10 mice were generated using each method. Post-operative mortality was zero with the Forceps and Pipette methods. However, 5 of 8 mice died within a day of implantation with the Needle method. At 6 weeks post-implantation, tumors were detectable in 80%, 67%, and 100% of surviving mice implanted using the Forceps, Needle, and Pipette methods, respectively. Tumor volumes were less variable with the Pipette compared to the Forceps method. Unlike the 100% engraftment for brain PDXs, the engraftment rate of patient brain metastasis implanted ectopically in MFP was only 75%. However, both orthotopic and ectopic PDXs grew about 2-fold faster after 3x serial passaging (all t test P < 0.05). To assess the effect of tumor site on drug sensitivity, we compared response of the brain and MFP PDXs (n = 5 each) to one intravenous dose of Epothilone B, a taxane-like drug that can pass through the blood-brain barrier. While this drug is effective against primary breast tumors, it failed to demonstrate activity against breast cancer brain metastasis tumors in a recent phase II clinical trial. Tumor progression of MFP PDXs was retarded by approximately 3-fold compared to control mice (P < 0.05). On the other hand, the drug had no effect on brain PDXs (P = 0.95). Our result implicate that the use of our model could have detected the ineffectiveness of this compound prior to clinical trial.
CONCLUSION: We established and characterized orthotopic breast cancer brain metastasis PDXs that were generated using a novel method that was facile, inexpensive, and efficient. PDXs created with our novel method may be powerful tools for preclinical studies of metastatic breast cancer.
Citation Format: Masanori Oshi, Maiko Okano, Kazuaki Takabe. Novel patient-derived orthotopic xenograft model for preclinical studies of breast cancer brain metastasis abstract. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 423.
Background
Incisional hernia (IH) is a common complication after colorectal surgery. However, the risk factors for incisional hernia after laparoscopic colorectal surgery (LCRS) have not been fully ...elucidated. This retrospective study analyzed the incidence rate of IH and evaluated the risk factors for IH after LCRS.
Methods
This was a retrospective multi-institution study of 423 colorectal cancer patients conducted between September 2012 and December 2014 in Yokohama Clinical Oncology Group. The diagnosis of IH was based on computed tomography and physical examination findings. The patient-, tumor-, and surgery-related variables were examined by univariate and multivariate analyses.
Results
A total of 423 patients were analyzed. The median follow-up period was 48.4 months. IH was observed in 36 patients (8.5%). The 1-year incidence of IH was 5.2%, and the 4-year incidence was 8.5%. A multivariate analysis showed that preoperative umbilical hernia (odds ratio OR 5.71; 95% confidence interval CI 2.02–16.10;
p
= 0.001) and a visceral fat area (VFA) ≥ 100 cm
2
(OR 2.74; 95% CI 1.08–6.96;
p
= 0.035) were independent risk factors of IH after LCRS.
Conclusions
The risk factors of IH after LCRS were preoperative umbilical hernia and VFA ≥ 100 cm
2
. In the case with an umbilical hernia or VFA ≥ 100 performing LCRS, it should likely NOT have a peri-umbilical extraction site and should be considered for an alternate site like a low transverse or Pfannenstiel incision.
Clinical Trials Registration:
The trial was registered with the UMIN Clinical Trials Registry, number 000038707.
E2F transcription factors play critical roles in the cell cycle. Therefore, their activity is expected to reflect tumor aggressiveness and responsiveness to therapy. We scored 3905 tumors of nine ...breast cancer cohorts for this activity based on their gene expression for the Hallmark E2F targets gene set. As expected, tumors with a high score had an increased expression of cell proliferation-related genes. A high score was significantly associated with shorter patient survival, greater MKI67 expression, histological grade, stage, and genomic aberrations. Furthermore, metastatic tumors had higher E2F scores than the primary tumors from which they arose. Although tumors with a high score had greater infiltration by both pro- and anti-cancerous immune cells, they had an increased expression of immune checkpoint genes. Estrogen receptor (ER)-positive/human epidermal growth factor receptor 2 (HER2)-negative cancer with a high E2F score achieved a significantly higher pathological complete response (pCR) rate to neoadjuvant chemotherapy. The E2F score was significantly associated with the expression of cyclin-dependent kinase (CDK)-related genes and strongly correlated with sensitivity to CDK inhibition in cell lines. In conclusion, the E2F score is a marker of breast cancer aggressiveness and predicts the responsiveness of ER-positive/HER2-negative patients to neoadjuvant chemotherapy and possibly to CDK and immune checkpoint inhibitors.
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