Postsurgical Survival Research Articles

P13.08 1H-MRS metabolomics as a predictor of overall survival in glioma patients

Abstract BACKGROUND We assess the efficacy of the metabolomic profile from glioma biopsies, in providing estimates of postsurgical Overall Survival in glioma patients. MATERIAL AND METHODS Tumor biopsies from 46 patients bearing gliomas, obtained neurosurgically in the period 1992–1998, were analyzed by high resolution 1H magnetic resonance spectroscopy (HR- 1H MRS), following retrospectively individual postsurgical Overall Survival up to 720 weeks. RESULTS The Overall Survival profile could be resolved in three groups; Short (shorter than 52 weeks, n=19), Intermediate (between 53 and 364 weeks, n=19) or Long (longer than 365 weeks, n=8), respectively. Classical histopathological analysis assigned WHO grades II-IV to every biopsy but notably, some patients with low grade glioma depicted unexpectedly Short Overall Survival, while some patients with high grade glioma, presented unpredictably Long Overall Survival. To explore reasons underlying this behavior, we analyzed HR- 1H MRS spectra from acid extracts of these biopsies, to identify the metabolite patterns underlying OS predictions. Poor prognosis was found in biopsies with higher contents of alanine, acetate, glutamate, total choline, phosphorylcholine and glycine, while more favorable prognosis was achieved in biopsies with larger contents of total creatine, glycerol-phosphorylcholine and myo-inositol. We implemented then a multivariate analysis approach to identify hierarchically the influence of these metabolomic biomarkers on OS predictions, using Classification Regression Trees (CRTs). Metabolomic CRTs grew up to 3 branches and split into 8 nodes, predicting correctly the outcome of 94.7% of the patients in the Short Overall Survival group, 78.9 % of the patients in the Intermediate Overall Survival group, and 75% of the patients in the Long Overall Survival group, respectively. CONCLUSION Present results suggest that metabolic profiling by HR-1H MRS provides more accurate Overall Survival estimates of glioma patients than classical histopathological grading, thus allowing to implement more accurate therapeutic decisions.

Science, Medicine, and the Anesthesiologist

Science, Medicine, and the Anesthesiologist

Clinicopathologic and epidemiological characteristics of prognostic factors in post-surgical survival of colorectal cancer patients in Jiangsu Province, China

Poor survival among colorectal cancer (CRC) patients has been widely associated with clinico-epidemiological features and treatment regimen. In Jiangsu (China), however, it is not known which one of the prognostic factors explains the survival disparities among patients with CRC. This prospective study using 1078 patients (stages I-IV) that underwent surgery at Jiangsu Hospital, explored the relevant factors affecting the prognoses of right-side colon cancer (RCC), left-side colon cancer (LCC) and rectal cancer (ReC) patients. Of these cases, 234 (21.7%), 241 (22.4%) and 603 (55.9%) were found to have RCC, LCC and ReC respectively. Compared to LCC, RCC exhibited a greater proportion of older patients, poorly differentiated carcinomas, higher T-stage and higher TNM-stage. The overall survival (OS) for RCC was 60 vs.78 or 77 months for LCC or ReC respectively (P = 0.030). There were no significant differences in OS between LCC and ReC across the subgroups (P = 0.633). In multivariate analysis, RCC patients had age (>60 vs. ≤60 years, HR = 1.529, P = 0.019), N-stage (N1 vs. N0, HR = 4.056, P = 0.012) and M-stage (M1 vs. M0, HR = 3.442, P < 0.0001) as independent prognostic factors, whereas smoking status was found to be a predictor of mortality (smoker vs. nonsmoker, HR = 2.343, P = 0.017) for LCC. In addition, age (>60 vs. ≤60 years, HR = 2.199, P < 0.0001), alcohol consumption (drinker vs. nondrinker, HR = 0.510, P = 0.034), tumor grade (Poor vs. well/moderate, HR = 2.759, P = 0.031) and T-stage (T3-4 vs. T1-2, HR = 1.742, P < 0.0001) were found to be predictors of mortality for ReC. There were significant pairwise interactions across subgroups. Furthermore, significant differences were observed for LCC vs. RCC (OS, HR = 0.783, P = 0.039), but no statistically significant differences for ReC vs. RCC (P = 0.149) and LCC vs. ReC (P = 0.355). Nevertheless, significant differences remained between ReC vs. RCC for male (HR = 0.591, P = 0.009), drinker (HR = 0.396, P = 0.005), rural resident (HR = 0.437,P = 0.022), tumor grade (well/moderate, HR = 0.475, P = 0.022), T-stage (T1-2, HR = 0.362, P = 0.001), N-stage (N0, HR = 0.604, P = 0.011), M-stage(M0, HR = 0.401, P = 0.006) and TNM-stage (I-II, HR = 0.567, P = 0.005). Statistically significant differences were observed for LCC vs. RCC for gender (female, HR = 0.495, P = 0.003) and T-stage (T1-2, HR = 0.417, P = 0.010) as well as for LCC vs. ReC in patients with smoking habits (HR = 1.951, P = 0.002) and M-stage (M0, HR = 2.291, P = 0.003). These findings suggest that the variations in CRC post-surgical survival in China may be primarily explained with the clinicopathologic features and epidemiological characteristic of the patients. Patients with RCC had significantly worse OS compared to both LCC and ReC in several subgroups.

Abstract 4620: Development of orthotopic breast cancer brain metastasis patient-derived xenograft(PDX) model

Abstract Background: The vast majority of the mortality of breast cancer results from distant metastasis and pre-clinical models are in critical need for effective drug development. Patient-derived xenograft (PDX) maintains the features of the donor tumors such as intra-tumor heterogeneity; however, PDX model of breast cancer brain metastasis that delivers stable consistent results is not popularized. We demonstrate our novel methods (coined “Oshi”method) used to develop an orthotopic brain metastasis patient-derived xenograft model (PDMOX) for breast cancer brain metastasis via tumor tissue implantation. Methods: Brain metastasis PDX were created using metastatic brain tumors from breast cancer patients and implanting them in the brain of NSG female mice. Tumors of ~1 mm3 were implanted mechanically minced tissue with medium. We implant the tissue through a frontal bone burr hole into the right caudate putamen. Tumor growth was monitored by magnetic resonance imaging (MRI). Results: “Oshi” method utilizes a pipette tip in order to inoculate tumor at the same depth. A minced tumor was instilled using a 23G needle as the “non-oshi” method. One hour post-surgical survival after implantation of minced tumor by “non-oshi” method was only 37.5% (3/8), whereas 100% (40/40) by the “Oshi” method. All tumors were well engrafted in surviving mice after both methods. There was larger variation in tumor growth after “non-oshi” (Median 20±25.0 day, range: 12-24 day. Tumor volume: median 5.6±21.0 mm3, range 2.8-48.7 mm3). One mouse developed ptosis, and 2 out of 3 mice that underwent the “non-oshi” method had sudden death. On the other hand, all mice that underwent the “Oshi” method lived until the tumor grew to 125-200 mm3 without neurological symptoms. The engraftment rate of Mincing tumor were 100% at 1.5 months after implantation, while the engraftment rate of Enzymatic digestion tumors was only 50% at 2 months. Mincing tumor had faster growth speed than the tumor of enzymatic digestion. The engraftment time of enzymatic digestion tumor was slower than other tumor tissue forms. All 3rd generation tumors could be detected at the point of 1month after implantation, on the other hand, it took 2 month to detect 1st generation tumors. The growth of 3rd generation tumors was significantly faster and bigger than others. Conclusion: Various surgical techniques used to generate PDMOX breast cancer models showed major differences in the tumors and outcomes. These novel models are expected to become powerful tools for preclinical studies in metastatic breast cancer. Citation Format: Masanori Oshi. Development of orthotopic breast cancer brain metastasis patient-derived xenograft(PDX) model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4620.

Assessment of Overall Survival in Glioma Patients as Predicted by Metabolomic Criteria.

Objective: We assess the efficacy of the metabolomic profile from glioma biopsies in providing estimates of postsurgical Overall Survival in glioma patients.Methods: Tumor biopsies from 46 patients bearing gliomas, obtained neurosurgically in the period 1992–1998, were analyzed by high resolution 1H magnetic resonance spectroscopy (HR- 1H MRS), following retrospectively individual postsurgical Overall Survival up to 720 weeks.Results: The Overall Survival profile could be resolved in three groups; Short (shorter than 52 weeks, n = 19), Intermediate (between 53 and 364 weeks, n = 19) or Long (longer than 365 weeks, n = 8), respectively. Classical histopathological analysis assigned WHO grades II–IV to every biopsy but notably, some patients with low grade glioma depicted unexpectedly Short Overall Survival, while some patients with high grade glioma, presented unpredictably Long Overall Survival. To explore the reasons underlying these different responses, we analyzed HR-1H MRS spectra from acid extracts of the same biopsies, to characterize the metabolite patterns associated to OS predictions. Poor prognosis was found in biopsies with higher contents of alanine, acetate, glutamate, total choline, phosphorylcholine, and glycine, while more favorable prognosis was achieved in biopsies with larger contents of total creatine, glycerol-phosphorylcholine, and myo-inositol. We then implemented a multivariate analysis to identify hierarchically the influence of metabolomic biomarkers on OS predictions, using a Classification Regression Tree (CRT) approach. The CRT based in metabolomic biomarkers grew up to three branches and split into eight nodes, predicting correctly the outcome of 94.7% of the patients in the Short Overall Survival group, 78.9% of the patients in the Intermediate Overall Survival group, and 75% of the patients in the Long Overall Survival group, respectively.Conclusion: Present results indicate that metabolic profiling by HR-1H MRS improves the Overall Survival predictions derived exclusively from classical histopathological gradings, thus favoring more precise therapeutic decisions.

INTRANASAL ADMINISTRATION OF MIDAZOLAM HYDROCHLORIDE IMPROVES SURVIVAL IN FEMALE SURF SCOTERS (MELANITTA PERSPICILLATA) SURGICALLY IMPLANTED WITH INTRACOELOMIC TRANSMITTERS.

The intracoelomic implantation of satellite transmitters is associated with lower survival in surf scoters (Melanitta perspicillata) compared with other species of diving ducks, potentially due to physiologic alterations following physical exertion and stress caused by handling and confinement. The effect of intranasal administration of midazolam hydrochloride on survival of surf scoters surgically implanted with intracelomic transmitters was evaluated. Shortly after their capture in Forestville (QC, Canada) in the fall of 2013, 26 randomly selected adult female surf scoters were administered midazolam hydrochloride (4.6-5.9 mg/kg) intranasally. The same volume of saline (1 mL) was given to another 26 adult female surf scoters as a control group. All birds were surgically implanted with an intracoelomic transmitter equipped with a percutaneous antenna. Transmitters were programmed to transmit 2 hr each day for 30 days after implantation, and mortality was estimated for each group using the telemetry data. The association between the administration of midazolam and survival was assessed while controlling for other factors such as body mass, transmitter-mass-to-body-mass ratio, hematocrit, total solids, and duration of surgery, anesthesia, and confinement. The odds of presumed death in the saline group were 5.3 times higher than in the midazolam group (95% confidence interval: 1.7, 19.0; P = 0.004). The presumed mortality at 30 days for the midazolam group (23%) was lower than for the saline group (61%). No other variable was significantly associated with survival. These results suggest that sedation with midazolam shortly after capture increased the postsurgical survival of female surf scoters surgically implanted with intracoelomic transmitters.

Prognostic significance of microRNAs in survival of patients with supratentorial gliomas

Aim. To identify novel microRNA markers as survival predictors in patients with supratentorial gliomas.Methods. This study involved the analysis of tumour and normal brain tissue biopsy samples obtained from patients undergoing combination treatment for supratentorial gliomas of different World Health Organization (WHO) grades. Real-time polymerase chain reaction was used to determine the expression profiles of ten microRNAs, following comparison with clinical treatment results: tumour morphology, WHO grade, patient age, Karnofsky scale, treatment type, postsurgical survival rate and histological diagnosis. The mean age of surgically treated patients [62 (57.9%) males and 45 (42.1%) females] was 48.8 ± 14 years. There were 17 (16%), 30 (28%) and 60 (56%) patients with grade II, III and IV (glioblastoma) gliomas, respectively. Statistical analysis was performed using Statistica version 10.0 and GraphPad Prism version 5.Results. Four microRNAs (miRNA-31, miRNA-21, miRNA-223 and miRNA-221) were strongly correlated with worse survival, when over-expressed, indicating their potential utility as survival predictors in glioma patients. Overexpression of these microRNAs in glioma tissue, lack of adjuvant therapy such as chemotherapy or radiotherapy and age &gt; 48 years were identified as factors for worse prognosis. Received 14 May 2019. Revised 21 June 2019. Accepted 25 June 2019. Funding: This work was supported by the program of fundamental scientific research on the topic 0310-2019-0003. Conflict of interest: The authors declare no conflict of interest.

Abstract P6-03-04: Successful development of patient-derived orthotopic xenograft models of brain and lung metastases of human breast cancer

Abstract Background: Metastatic breast cancer is the reason that we continue to lose 40,000 women every year in the US. Without appropriate pre-clinical model, success rate of clinical trials continue to suffer. Because syngeneic mouse models utilize murine neoplasm that may not represent human cancer, patient-derived xenografts (PDX) have emerged as a pre-clinical model that maintains human cancer features such as intratumoral heterogeneity. However, there is no established orthotopic PDX model for metastatic breast cancer even though the main cause of death is brain and lung metastasis. Orthotopic brain or lung PDX is expected to reproduce the original tumor microenvironment. We describe our new patient-derived metastasis orthotopic xenograft (PDMOX) mouse models of human breast cancer. Methods: All work was performed in female NSG mice of age 8-12 m. Breast cancer metastatic tumors from brain and lung that had been passaged 3x in mammary fat pads were used. Tumors of 1 mm3 were implanted orthotopically in two forms: solid piece, or minced tissue with 3 μl Matrigel. Tumor growth was monitored by MRI. Results: Two methods for brain PDMOX were compared. “Manual push” method implanted minced tumor through a frontal bone burr hole into right caudate putamen at 4 mm depth using forceps. “Pipette tip” method utilized either a pipetter for minced tissue or Hamilton syringe for solid tissue to inoculate tumor. One hour post-surgical survival was 37.5% (3/8) after “manual push”, and 100% (30/30) after “Pipette tip” method. All tumors engrafted in surviving mice with either method. However, the tumors formed on brain surface and parenchyma invasion was rare after “manual push” method, whereas solid tumor invaded parenchyma by “pipette tip” method. Therefore, it was no surprise to find large variation in tumor growth after “Manual push” (detection time 17±5.0 d, range: 17-26; volume 5.6±21.0 mm3, range 2.8-48.7). One mouse developed ptosis, and 2 out of 3 mice that underwent “Manual push” had sudden death. On the other hand, all mice that underwent “Pipette tip” method lived until tumor grew to 125-200 mm3, without neurological symptoms. These brain tumors could be passaged with 100% success (9/9). For right lung PDMOX, “thoracotomy” and “non-thoracotomy” methods were compared. “Thoracotomy” method implanted a solid tissue using forceps or 8-0 nylon suture, or injected minced tissue 1 mm below pleura. “Non-thoracotomy” method injected minced tissue using 23G needle. One hour post-surgical survival was 30% (9/30, 8/30) after “thoracotomy” method using forceps or suture, resp. However, survival using suture method could be significantly improved to 97% (29/30) by reducing thoracotomy length (&amp;lt;10 vs. ≥10 mm: t test P = 0.003). Post-operative survival was not affected by age, weight, or operation or anesthesia time. On the other hand, all mice after “non-thoracotomy” method survived, but chest wall implantation occurred in 67% (4/6) when the method was performed using a cell line. Conclusion: By simple modifications of surgical techniques, we could establish orthotopic brain and lung xenografts of breast cancer tumors with almost zero mortality and 100% engraftment. Our novel PDMOX models can be powerful tools for preclinical studies. Citation Format: Oshi M, Okano M, Takabe K. Successful development of patient-derived orthotopic xenograft models of brain and lung metastases of human breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P6-03-04.

ASO Author Reflections: Tumor-Infiltrating NETs are New Biomarkers to Predict Postsurgical Survival for Patients with Pancreatic Ductal Adenocarcinoma

ASO Author Reflections: Tumor-Infiltrating NETs are New Biomarkers to Predict Postsurgical Survival for Patients with Pancreatic Ductal Adenocarcinoma

Intratumoral collagen index predicts mortality and survival in canine cutaneous mast cell tumours.

Mast cell tumours (MCTs) constitute almost 25% of cutaneous neoplasms in dogs. Their biological behaviour is predicted using histopathological grading which is based on several subjective criteria that are vulnerable to intra- and interobserver variability. To improve the prediction of the biological behaviour, several complementary markers have been studied. The integrity of the extracellular matrix (ECM) may play a protective role against tumoral progression, and favour cellular proliferation, angiogenesis, invasion and metastases when altered. To evaluate the quantification of collagen and elastic fibres as prognostic markers for MCTs. Thirty-eight random cases of canine cutaneous MCT surgically treated with wide margins were included. Intratumoral collagen and elastic fibres were identified and quantified on histological sections stained with Masson's trichrome, Picrosirius red and Verhoeff; the results were compared with histopathological grades, mortality due to the disease and postsurgical survival. Morphometric analysis revealed a significant relationship between histopathological grade and intratumoral collagen index (CoI). In addition, the CoI was considered an independent indicator for mortality and postsurgical survival. These results support the importance of the CoI in the grading and prognosis of MCTs, suggesting that preservation and/or synthesis of collagen have the potential to become targets for MCT therapeutics.

Long-Term Implications of Atrial Fibrillation in Patients With Degenerative Mitral Regurgitation

BackgroundScientific guidelines consider atrial fibrillation (AF) complicating degenerative mitral regurgitation (DMR) a debated indication for surgery. ObjectivesThis study analyzed the prognostic/therapeutic implications of AF at DMR diagnosis and long-term. MethodsPatients were enrolled in the MIDA (Mitral Regurgitation International Database) registry, which reported the consecutive, multicenter, international experience with DMR due to flail leaflets echocardiographically diagnosed. ResultsAmong 2,425 patients (age 67 ± 13 years; 71% male, 67% asymptomatic, ejection fraction 64 ± 10%), 1,646 presented at diagnosis with sinus rhythm (SR), 317 with paroxysmal AD, and 462 with persistent AF. Underlying clinical/instrumental characteristics progressively worsened from SR to paroxysmal to persistent AF. During follow-up, paroxysmal and persistent AF were associated with excess mortality (10-year survival in SR and in paroxysmal and persistent AF was 74 ± 1%, 59 ± 3%, and 46 ± 2%, respectively; p < 0.0001), that persisted 20 years post-diagnosis and independently of all baseline characteristics (p values <0.0001). Surgery (n = 1,889, repair 88%) was associated with better survival versus medical management, regardless of all baseline characteristics and rhythm (adjusted hazard ratio: 0.26; 95% confidence interval: 0.23 to 0.30; p < 0.0001) but post-surgical outcome remained affected by AF (10-year post-surgical survival in SR and in paroxysmal and persistent AF was 82 ± 1%, 70 ± 4%, and 57 ± 3%, respectively; p < 0.0001). ConclusionsAF is a frequent occurrence at DMR diagnosis. Although AF is associated with older age and more severe presentation of DMR, it is independently associated with excess mortality long-term after diagnosis. Surgery is followed by improved survival in each cardiac rhythm subset, but persistence of excess risk is observed for each type of AF. Our study indicates that detection of AF, even paroxysmal, should trigger prompt consideration for surgery.

TLX3 repressed SNAI1-induced epithelial-mesenchymal transition by directly constraining STAT3 phosphorylation and functionally sensitized 5-FU chemotherapy in hepatocellular carcinoma.

TLX3 has an established role as a sequence-specific transcription factor with vital functions in the nervous system. Although several studies have shown that TLX3 is aberrantly up-regulated in leukemia, its expression and function in hepatocellular carcinoma (HCC) remain unknown. We found that TLX3 expression was decreased in 68/100 (68%) HCC cases and negatively correlated with the expression of p-STAT3, SNAI1, and Vimentin, while it was positively associated with E-cadherin expression. ITRAQ proteomic profiling revealed significantly less TLX3 expression in primary HCC tumors than in portal vein tumor thrombi. Comparison of Kaplan-Meier curves showed that down-regulation of TLX3 in HCC was associated with poor post-surgical survival. TLX3 over-expression inhibited HCC cell viability, proliferation, migration, invasion and enhanced 5-FU treatment, whereas silencing TLX3 produced the opposite results. Further experiments showed that TLX3 attenuated the EMT phenotype. In vivo experiments showed that knockdown of TLX3 promoted the growth of HCC xenografts and attenuated the anti-tumor effects of 5-FU treatment. Gene expression microarray analysis revealed that TLX3 inhibited IL-6/STAT3 signaling. In additional mechanistic studies TLX3 reversed the EMT phenotype of HCC cells by binding to STAT3, inhibiting STAT3 phosphorylation, and down-regulating SNAI1 expression. Taken together, loss of expression of TLX3 induces EMT by enhancing IL-6/STAT3/SNAI1 signaling, and accelerates HCC progression while also attenuated the effect of 5-FU on HCCs.

Tumor-based gene expression biomarkers to predict survival following curative intent resection for stage I lung adenocarcinoma

BackgroundPrognostic biomarkers are needed in clinical setting to predict outcome after resection for early-stage lung adenocarcinoma. The goal of this study is to validate tumor-based single-gene expression biomarkers with demonstrated prognostic value in order to move them along the clinical translation pipeline.MethodsPrognostic genes were selected from the literature and the best candidates measured by quantitative real-time polymerase chain reaction (qPCR) in tumors of 233 patients with stage I adenocarcinoma. Significant prognostic genes were then validated in an independent set of 210 patients matching the first set in terms of histology, stage, and clinical data.ResultsEleven genes with demonstrated prognostic value were selected from the literature. Complementary analyses in public databases and our own microarray dataset led to the investigation of six genes associated with good (BTG2, SELENBP1 and NFIB) or poor outcome (RRM1, EZH2 and FOXM1). In the first set of patients, EZH2 and RRM1 were significantly associated with better survival on top of age, sex and pathological stage (EZH2 p = 3.2e-02, RRM1 p = 5.9e-04). The prognostic values of EZH2 and RRM1 were not replicated in the second set of patients. A trend was observed for both genes in the joint analyses (n = 443) with higher expression associated with worse outcome.ConclusionAdenocarcinoma-specific mRNA expression levels of EZH2 and RRM1 are associated with poor post-surgical survival in the first set of patients, but not replicated in a clinically and pathologically matched independent validation set. This study highlights challenges associated with clinical translation of prognostic biomarkers.

HHLA2 is a novel immune checkpoint protein in pancreatic ductal adenocarcinoma and predicts post-surgical survival

HHLA2 is a newly identified member of the B7 immune checkpoint family, but its function and crosstalk with immune cells is not fully understood. To gain insights into the HHLA2 expression profile and to determine the clinical significance and function of HHLA2 in pancreatic cancer, we performed immunohistochemistry (IHC) analyses on tissue microarrays (TMAs) of pancreatic ductal adenocarcinoma (PDAC, n = 92) with matched peritumoral tissues as well as in cohorts of precancerous lesions: pancreatic intraepithelial neoplasia (PanIN) and intraductal papillary mucinous neoplasm (IPMN). We found that HHLA2 was rarely detected in normal acinar, islet, and ductal cells but widely expressed from early pancreatic precancerous lesions to invasive PDAC. The overall HHLA2 positivity was 95% (19/20) in low grade PanIN and 70.73% (29/41) in IPMN. HHLA2 expression was detected in 77.17% (71/92) of the PDAC cases and was significantly associated with better prognosis in this cohort. Our findings suggest that HHLA2 may behave as a costimulatory ligand in pancreatic cancer, which differs from other B7 family members that are largely characterized as checkpoint inhibitors. Further investigation of the HHLA2 signaling pathway and its receptors is warranted by our data and may lead to novel therapeutic interventions.

Diacylglycerol kinase γ predicts prognosis and functions as a tumor suppressor by negatively regulating glucose transporter 1 in hepatocellular carcinoma

Diacylglycerol kinases (DGK) are a family of enzymes catalyzing the transformation of diacylglycerol into phosphatidic acid, which have been recognized as key regulators in cell signaling pathways. The role of DGKγ in human malignancies has seldom been studied. In this study, we investigated the role of DGKγ in hepatocellular carcinoma (HCC). We found that DGKγ was down-regulated in HCC tumor tissues and cell lines as compared to that in non-tumor tissues. The prognostic value of DGKγ expression was evaluated by Cox regression and Kaplan-Meier analyses. Lower DGKγ expression in tumor tissues was an independent prognostic factor for poor post-surgical overall survival. By using HDACs inhibitors treatment and ChIP-PCR, we discovered that histone H3 and H4 deacetylation mainly contributed to the downregulation of DGKγ expression. Functional studies revealed that ectopic expression of DGKγ inhibited cell proliferation and cell migration in HCC cells. Mechanism studies showed that DGKγ overexpression led to down regulation of GLUT1 protein level and AMPK activity, which result in glucose uptake suppression as well as lactate and ATP production declination. The decrease of GLUT1 level could be partially rescued by treatments with either DGK inhibitor and lysosome inhibitor, indicating DGKγ may down-regulate GLUT1 through its kinase activity and lysosome degradation process. Together, this study demonstrated that DGKγ plays a tumor suppressor role in HCC by negatively regulating GLUT1. DGKγ could be a novel prognostic indicator and therapeutic target for HCC.

Should minimally invasive lung adenocarcinoma be transferred from stage IA1 to stage 0 in future updates of the TNM staging system?

The 8th International Association Study of Lung Cancer (IASLC) TNM classification staging project for lung cancer has classified patients with adenocarcinoma in situ (AIS) into stage 0, while patients with a minimally invasive adenocarcinoma (MIA) were classified into stage IA1. However, MIA patients, similar to AIS patients, have an approximately a 100% 5-year survival. This study aimed to investigate if MIA could be transferred from stage IA to stage 0 in the next TNM staging system. We retrospectively reviewed 1,524 consecutive patients with a pathologically confirmed AIS, MIA and an invasive adenocarcinoma (IADC) in stage IA1. Disease-free survival (DFS) and overall survival (OS) were analyzed to evaluate survival difference between these three groups. There were 412 AIS, 675 MIA and 437 IADC patients in stage IA1. No statistically significant differences for DFS and OS (P=0.109) were seen between the AIS and MIA groups. Patients of the IADC group had significantly worse DFS (P=0.003) but the OS rate (P=0.941) was insignificant when compared with the MIA patients. Similar survival results were seen when comparisons were made between the IADC and AIS/MIA groups. The IADC group had a worse DFS (P=0.001) rate but no OS (P=0.380) difference with the AIS/MIA groups. Patients with AIS and MIA had similar post-surgical survival rates. We propose that MIA may potentially be transferred from IA1 to stage 0 in the future.

Tumor-Infiltrating NETs Predict Postsurgical Survival in Patients with Pancreatic Ductal Adenocarcinoma.

Tumor-infiltrating neutrophils (TINs) indicate poor prognosis for patients with pancreatic ductal adenocarcinoma (PDAC). Activated neutrophils can generate neutrophil extracellular traps (NETs). Little is known about the presence and prognostic significance of tumor-infiltrating NETs in PDAC. This study enrolled 317 patients, in two independent sets (training and validation), who underwent curative pancreatectomy for PDAC in Shanghai Cancer Center. TINs and NETs were identified by immunohistochemical staining for CD15 and citrullinated histone H3, respectively. The relationship between clinicopathological features and outcomes was analyzed. Accuracy of prognostic prediction models was evaluated using concordance index (C-index) and Akaike information criterion (AIC). NETs were associated with OS (both, P < 0.001) and RFS (both, P < 0.001) in the training and validation sets. Tumor-infiltrating NETs predicted poor postsurgical survival of patients with PDAC. Moreover, multivariate analysis identified NETs and AJCC TNM stage as two independent prognostic factors for OS and RFS. Combination of NETs with the 8th edition TNM staging system (C-index, 0.6994 and 0.6669, respectively; AIC, 1067 and 1126, respectively) generated a novel model that improved the predictive accuracy for survival in both sets (C-index, 0.7254 and 0.7117, respectively; AIC, 1047 and 1102, respectively). The model combining presence of NETs with the 7th edition AJCC TNM staging system also had improved predictive accuracy. NETs were an independent prognostic factor in PDAC and incorporation of NETs along with the standard TNM stating system refined risk-stratification and predicted survival in PDAC with improved accuracy.

ASO Author Reflections: Tumor-Infiltrating Platelets Predict Postsurgical Survival in Patients with Pancreatic Ductal Adenocarcinoma.

ASO Author Reflections: Tumor-Infiltrating Platelets Predict Postsurgical Survival in Patients with Pancreatic Ductal Adenocarcinoma.

Tumor-Infiltrating Platelets Predict Postsurgical Survival in Patients with Pancreatic Ductal Adenocarcinoma

Platelets are believed to promote tumor growth and metastasis in several tumor types. The prognostic role of blood platelets in pancreatic ductal adenocarcinoma (PDAC) remains controversial, and the prognostic value of tumor-infiltrating platelets (TIPs) remains unknown. A total of 303 patients who underwent curative pancreatectomy for PDAC were enrolled from two independent centers in China and divided into three cohorts. Paired preoperative blood samples and surgical specimens from all patients were analyzed. The correlations between patient outcomes and preoperative blood platelet counts and the presence of TIPs, respectively, were analyzed. TIPs were identified by immunohistochemical staining of CD42b. Prognostic accuracy was estimated by concordance index (C-index) and Akaike information criterion (AIC). TIPs, but not preoperative blood platelet counts, were associated with overall survival (OS; all P < 0.001) and recurrence-free survival (RFS; all P < 0.001) in the training, testing, and validation sets. Positive CD42b expression predicted poor postsurgical survival. Incorporation of TIPs improved the predictive accuracy of the 8th edition American Joint Committee on Cancer (AJCC) tumor-node-metastasis (TNM) staging system for OS in each of the three cohorts (C-index: 0.7164, 0.7569, and 0.7050, respectively; AIC: 472, 386, and 1019, respectively). The new predictor system was validated by incorporating TIPs with the 7th edition AJCC TNM staging system (C-index: 0.7052, 0.7623, and 0.7157; AIC: 476, 386, and 1015). TIPs were an independent prognostic factor that could be incorporated into the AJCC TNM staging system to refine risk stratification and predict surgical outcomes of patients with PDAC.

Androgen receptor drives hepatocellular carcinogenesis by activating enhancer of zeste homolog 2-mediated Wnt/β-catenin signaling.

BackgroundAndrogen receptor (AR) plays a crucial role as a transcription factor in promoting the development of hepatocellular carcinoma (HCC) which is prone to aberrant chromatin modifications. However, the regulatory effects of AR on epigenetic mediators in HCC remain ill-defined. Enhancer of zeste homolog 2 (EZH2), an oncogene responsible for the tri-methylation of histone H3 at lysine 27 (H3K27me3), was identified to be overexpressed in approximate 70–90% of HCC cases, which prompted us to investigate whether or how AR regulates EZH2 expression.MethodsColony formation, soft agar assay, xenograft and orthotopic mouse models were used to determine cell proliferation and tumorigenicity of gene-manipulated HCC cells. Gene regulation was assessed by chromatin immunoprecipitation, luciferase reporter assay, quantitative RT-PCR and immunoblotting. Clinical relevance of candidate proteins in patient specimens was examined in terms of pathological parameters and postsurgical survival rates.FindingsIn this study, we found that AR upregulated EZH2 expression by binding to EZH2 promoter and stimulating its transcriptional activity. EZH2 overexpression increased H3K27me3 levels and thereby silenced the expression of Wnt signal inhibitors, resulting in activation of Wnt/β-catenin signaling and subsequently induction of cell proliferation and tumorigenesis. In a cohort of human HCC patients, concordant overexpression of AR, EZH2, H3K27me3 and active β-catenin was observed in tumor tissues compared with paired non-tumor tissues, which correlated with tumor progression and poor prognosis. These findings demonstrate a novel working model in which EZH2 mediates AR-induced Wnt/β-catenin signaling activation through epigenetic modification, and support the application of EZH2-targeted reagents for treating HCC patients.