Methylation-associated silencing of TU3A in human cancers

TU3A, located on 3p21.1, was originally identified as a candidate tumor suppressor gene in renal cell carcinoma (RCC). Recently, down-regulation of TU3A expression has been reported not only in RCC but also in other types of cancers. However, no studies have evaluated the mechanism underlying TU3A inactivation. In the present study, we first examined the expression and promoter CpG island methylation of TU3A in RCC. TU3A mRNA was slightly or not expressed in 3 RCC cell lines (ACHN, Caki-1 and NC65). Bisulfite sequencing of the TU3A promoter and treatment of the RCC cell lines with 5-aza-2'-deoxycytidine and/or trichostatin A revealed an association between TU3A expression and promoter hypermethylation. Next, we analyzed TU3A methylation in primary RCC by using combined bisulfite restriction analysis. Mean methylated fraction was 19.2% (range: 0-57.3%) in 53 conventional RCCs and 2.3% (range: 0-12.7%) in 24 corresponding normal kidneys. We defined a methylation fraction of >20% as hypermethylation. TU3A hypermethylation was detected in 22 (41.5%) of 53 RCCs and significantly associated with advanced tumor stage (>T2 vs. T1 and T2: P=0.005, > or = N1 or M1 vs. N0M0: P=0.001) and poor disease-specific survival (P=0.0038). Furthermore, we observed promoter hyper-methylation of TU3A in several types of cancer cell lines and primary cancers of the bladder and testis. To our knowledge, the present study is the first to demonstrate the epigenetic inactivation of TU3A in human cancers. The findings of this study warrant further study to investigate the role of TU3A methylation in cancer development.     

A structural equation model relating oral condition,denture quality,chewing ability,satisfaction, and oral health-related quality of life in complete denture wearers

ObjectivesTo investigate the relationship between mandibular ridge form, stability and retention of mandibular complete denture, accuracy of jaw relation recording, patients’ perception of chewing ability, satisfaction with dentures and oral health-related quality of life (OHRQoL) in complete denture wearers.MethodsA total of 183 edentulous patients, who visited the Dental Hospital of Tokyo Medical and Dental University for new complete dentures, were recruited. Oral examination was performed. Cawood and Howell's method was used to grade the mandibular ridge form. The stability and retention of the mandibular complete denture were assessed using Kapur method. Accuracy of jaw relation recording was evaluated using a newly developed jaw relation index. Patients’ perception of chewing ability was rated using a food intake questionnaire. Patients’ satisfaction with complete dentures was assessed on a 100-mm visual analogue scale. OHRQoL was measured using the Japanese version of the Oral Health Impact Profile for edentulous subjects. A structural equation model was constructed based on the hypothesis that oral condition and denture quality would be related to chewing ability, satisfaction and OHRQoL.ResultsSignificant relationships were found between mandibular ridge form, stability of mandibular complete denture, accuracy of jaw relation recording, perceived chewing ability, satisfaction and OHRQoL. Various fit indices were within acceptable limits.ConclusionsOral condition and denture quality were related to patients’ perception of chewing ability, satisfaction with dentures and OHRQoL in complete denture wearers.Clinical significanceA favourable oral condition and denture quality are important for successful complete denture therapy.     

Synergistic Antitumor Interaction of Human Monocyte Chemotactant Protein-1 Gene Transfer and Modulator for Tumor-Infiltrating Macrophages

Purpose. In order to evaluate the possibility of synergistic antitumor gene therapy by the gene delivery of monocyte chemotactant protein-1 (MCP-1/MCAF/IE), the effect of a biological response modulater for macrophages on tumor progression of gene transfected tumor cells was studied. Methods. Cachexia-inducing adenocarcinoma cells (cell line colon 26, clone 20) were transfected with either a control plasmid or MCP-1 cDNA. Results. The production of MCP-1 reached 70-80 ng/ml in vitro when transfectant cells were cultured at a cell density of 1 × 10⁵ cells/ml for 3 days. Transfection of MCP-1 cDNA did not affect the growth ratein vitro. Also, MCP-1-transfectants formed tumors after intra-footpad inoculation similar in size to the parental cells. The number of infiltrating macrophages in the primary tumor of the transfectant rapidly increased from the 3rd to 5th day after inoculation as revealed by immunohistochemical staining using an antibody against mouse macrophages. An earlier, greater, but no longer-lasting increase in tumor-infiltrating macrophages was induced in tumors by MCP-1 transfection was compared to that induced by the parent cells. On the 10th day after the inoculation, the tumor-infiltrating macrophages in mice inoculated MCP-1 transfectants were decreased to a level similar to that of the parent cells. Groups of mice were treated intraperitoneally with LPS at different times after the inoculation. Tumor cells producing high levels of MCP-1 were significantly lysed by macrophages treated with LPS, whereas parental or control transfected cells were not. Conclusions. Combination immunotherapy can provide a rationale for the application of MCP-1 treatment to increase immunological responses to cancer.     

Genetic polymorphisms of the interleukin-4 receptor alpha gene are associated with an increasing risk and a poor prognosis of sporadic renal cell carcinoma in a Japanese population.

PURPOSE: It has been suggested that the immune system of the host may be capable of modulating the clinical course of renal cell carcinoma (RCC) patients. In fact, the amount of Th2 cytokines such as interleukin (IL)-4 and IL-10 in the serum of patients has been found to be an important predictor of poor prognosis. Recently, it was reported that genetic polymorphisms of the IL-4 receptor alpha (IL-4Ralpha) gene affect the strength of signaling through the receptor. In addition, these same polymorphisms were found to be associated with an increased risk of atopy by causing Th2-dominated responses of the host. The significance of the polymorphisms on the incidence and prognosis in sporadic RCC patients were examined to clarify the role of IL-4 as well as that of the Th1/Th2 immune system in this disease. EXPERIMENTAL DESIGN: A case-control study was performed with 143 sporadic RCCs in a Japanese population and 205 Japanese controls. Logistic regression models were also used to assess the genetic effects on prognosis. RESULTS: The frequencies of variant alleles that enhance signaling of IL-4 were significantly related to an increased risk of RCC. Furthermore, multivariate regression analysis showed that the genotype of the IL-4R gene was an independent prognostic factor for cause-specific survival (P = 0.018) together with M classification (P = 0.0002) and histopathological grade (P = 0.044). CONCLUSIONS: The present findings show that the preferential Th2-type response to tumors was associated with a poorer prognosis and suggest that polymorphisms of the IL-4Ralpha gene may serve as useful genetic markers for assessing the risk of the development and progression of RCC.     

Polygenic Risk Score of Adolescent Idiopathic Scoliosis for Potential Clinical Use

Adolescent idiopathic scoliosis (AIS) is a common disease causing three-dimensional spinal deformity in as many as 3% of adolescents. Development of a method that can accurately predict the onset and progression of AIS is an immediate need for clinical practice. Because the heritability of AIS is estimated as high as 87.5% in twin studies, prediction of its onset and progression based on genetic data is a promising option. We show the usefulness of polygenic risk score (PRS) for the prediction of onset and progression of AIS. We used AIS genomewide association study (GWAS) data comprising 79,211 subjects in three cohorts and constructed a PRS based on association statistics in a discovery set including 31,999 female subjects. After calibration using a validation data set, we applied the PRS to a test data set. By integrating functional annotations showing heritability enrichment in the selection of variants, the PRS demonstrated an association with AIS susceptibility (p = 3.5 × 10⁻⁴⁰ with area under the receiver-operating characteristic [AUROC] = 0.674, sensitivity = 0.644, and specificity = 0.622). The decile with the highest PRS showed an odds ratio of as high as 3.36 (p = 1.4 × 10⁻¹⁰) to develop AIS compared with the fifth in decile. The addition of a predictive model with only a single clinical parameter (body mass index) improved predictive ability for development of AIS (AUROC = 0.722, net reclassification improvement [NRI] 0.505 ± 0.054, p = 1.6 × 10⁻⁸), potentiating clinical use of the prediction model. Furthermore, we found the Cobb angle (CA), the severity measurement of AIS, to be a polygenic trait that showed a significant genetic correlation with AIS susceptibility (rg = 0.6, p = 3.0 × 10⁻⁴). The AIS PRS demonstrated a significant association with CA. These results indicate a shared polygenic architecture between onset and progression of AIS and the potential usefulness of PRS in clinical settings as a predictor to promote early intervention of AIS and avoid invasive surgery. © 2021 American Society for Bone and Mineral Research (ASBMR).     

CT-based morphological analysis of spinal fractures in patients with diffuse idiopathic skeletal hyperostosis

Purpose

To clarify correlations between spinal fracture and delayed paralysis in patients with diffuse idiopathic skeletal hyperostosis (DISH) using computed tomography (CT) with multiplanar reformatting (CT-MPR). DISH increases susceptibility to unstable spinal fractures, leading to neurological deterioration. The pathomechanism of the neurological injury is unclear.

Association of two-staged surgery with systemic perioperative complications in lateral lumbar interbody fusion for adult spinal deformity: a propensity score-weighted study

Purpose

Adult spinal deformity (ASD) surgery carries a higher risk of perioperative systemic complications. However, evidence for the effect of planned two-staged surgery on the incidence of perioperative systemic complications is scarce. Here, we evaluated the effect of two-staged surgery on perioperative complications following ASD surgery using lateral lumbar interbody fusion (LLIF).

Methods

The study was conducted under a retrospective multi-center cohort design. Data on 293 consecutive ASD patients (107 in the two-staged group and 186 in the one-day group) receiving corrective surgery using LLIF between 2012 and 2021 were collected. Clinical outcomes included occurrence of perioperative systemic complications, reoperation, and intraoperative complications, operation time, intraoperative blood loss, transfusion, and length of hospital stay. The analysis was conducted using propensity score (PS)-stabilized inverse probability treatment weighting to adjust for confounding factors. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated in a PS-weighted cohort.

Results

In this cohort, 19 (18.4%) patients in the two-staged group and 43 (23.1%) patients in the one-day group experienced any systemic perioperative complication within 30 days following ASD surgery. In the PS-weighted cohort, compared with the patients undergoing one-day surgery, no association with the risk of systemic perioperative complications was seen in patients undergoing two-staged surgery (PS-weighted OR 0.78, 95% CI 0.37–1.63; p = 0.51).

Conclusion

Our study suggested that two-staged surgery was not associated with risk for perioperative systemic complications following ASD surgery using LLIF.

     

Increased Akt and phosphorylated Akt expression are associated with malignant biological features of prostate cancer in Japanese men

OBJECTIVE: To investigate the relationship between the expression of Akt (a serine/threonine kinase that plays a central role in tumorigenesis), phosphorylated Akt (p-Akt), prostate cancer tumour grade, androgen receptor (AR)-staining score, and Ki67 labelling index (LI) in Japanese men. PATIENTS, MATERIALS AND METHODS: The expression and activation of the cell survival protein Akt was analysed by immunohistochemical staining of paraffin-embedded tissue microarray sections of prostate carcinoma taken from 52 Japanese men who under radical prostatectomy. The correlation between the expression of Akt and p-Akt, and their relationship to primary Gleason grade, AR expression and Ki-67 LI was investigated. RESULTS: The expression of Akt and p-Akt were positively related to primary Gleason grade (Fisher's exact test, P = 0.002 and P = 0.032, respectively). Both AR-staining score and Ki67 LI were were positively related to the expression of Akt (both P < 0.001) and p-Akt (P < 0.001 and P = 0.008, respectively). There was a significant positive correlation between the expression of Akt and p-Akt (Spearman's correlation, r = 0.644, P < 0.001). CONCLUSIONS: Increased expression of both Akt and p-Akt were associated with higher tumour grade as well as a higher AR-staining score and Ki67 LI. These data indicate that Akt and p-Akt might be molecular markers for detecting malignant biological features of prostate cancer in the Japanese population.