Establishment of patient-derived three-dimensional organoid culture in renal cell carcinoma

Purpose: Renal cell carcinoma is a heterogeneous kidney cancer, and over 403,000 cases were reported worldwide in 2018. Current methods for studying renal cell carcinoma are limited to two-dimensional (2D) culture of primary cell lines and patient-derived xenograft models. Numerous studies have suggested that 2D culture poorly represents the diversity, heterogeneity, and drug-resistance of primary tumors. The time and cost associated with patient-derived xenograft models poses a realistic barrier to their clinical utility. As a biomimetic model, patient-derived three-dimensional (3D) organoid culture can overcome these disadvantages and bridge the gap between in vitro cell culture and in vivo patient-derived xenograft models. Here, we establish a patient-derived 3D organoid culture system for clear cell renal cell carcinoma and demonstrate the biomimetic characteristics of our model with respect to both primary kidney cancer and conventional 2D culture. Materials and Methods: Normal renal tissues and tumor tissues were collected from patients with clear cell renal cell carcinoma. The dissociated cells were cultured as conventional 2D culture and 3D organoid culture. The biomimetic characteristic of the two cultures were compared. Results: Compared with 2D culture, the 3D organoid cultures retained the characteristic lipid-rich, clear cell morphology of clear cell renal cell carcinoma. Carbonic anhydrase 9 and vimentin were validated as biomarkers of renal cell carcinoma. Expression of the two validated biomarkers was more enhanced in 3D organoid culture. Conclusions: Patient-derived 3D organoid culture retains the characteristics of renal cell carcinoma with respect to morphology and biomarker expression.ope

Effect of intraoperative fluid volume on postoperative ileus after robot-assisted radical cystectomy

This study aimed to investigate the effect of intraoperative fluid volume on the postoperative ileus (POI) recovery period. A retrospective review of the Korean robot-assisted radical cystectomy database identified 718 patients who underwent robot-assisted radical cystectomy (RARC). Regression analyses were performed to identify the associations between the amount of intraoperative fluid administration (crystalloid/colloid/total), POI period (time to flatus/bowel movements), and length of hospital stay (LOS) after adjusting for covariates. In addition, we analyzed the risk factors for gastrointestinal complications and prolonged POI using a logistic regression model. An increasing volume of the administered crystalloid/total fluid was associated with prolonged POI (crystalloid R2 = 0.0725 and P < 0.0001; total amount R2 = 0.0812 and P < 0.0001), and the total fluid volume was positively associated with the LOS (R2 = 0.099 and P < 0.0001). The crystalloid amount was a risk factor for prolonged POI (P < 0.001; odds ratio, 1.361; 95% confidence interval, 1.133-1.641; P < 0.001). In the context of RARC, increased intravenous fluids are associated with prolonged POI and longer LOS.ope

Diagnostic accuracy of prostate-specific antigen below 4 ng/mL as a cutoff for diagnosing prostate cancer in a hospital setting: A systematic review and meta-analysis

Purpose: A prostate-specific antigen (PSA) cutoff of 4 ng/mL has been widely used for prostate cancer screening in population-based settings. However, the accuracy of PSA below 4 ng/mL as a cutoff for diagnosing prostate cancer in a hospital setting is inconclusive. We systematically reviewed the accuracy of PSA below 4 ng/mL cutoff in a hospital setting. Materials and methods: We systematically reviewed the literature by searching major databases until March 2020, and a meta-analysis and quality assessment were performed. Results: A total of 11 studies were included at the completion of the screening process. The meta-analysis showed a sensitivity of 0.92 and a specificity of 0.16 for a PSA cutoff below 4 ng/mL. The area under the hierarchical summary receiver operating characteristic curve was 0.87, the positive likelihood ratio was 1.23, the negative likelihood ratio was 0.46, and the diagnostic odds ratio was 2.64. PSA sensitivities and specificities varied according to the cutoff range: 0.94 and 0.17 for 2 to 2.99 ng/mL, and 0.92 and 0.16 for 3 to 3.99 ng/mL, respectively. No significant differences in the sensitivity and specificity of PSA cutoffs in the range of 2 to 2.99 ng/mL and 3 to 3.99 ng/mL were found. Conclusions: Although a PSA cutoff <3 ng/mL is relatively more sensitive and specific than PSA ≥3 ng/mL, no significant differences in sensitivity and specificity were found in the diagnosis of prostate cancer. Therefore, clinicians should choose an appropriate PSA cutoff on the basis of clinical circumstances and patients' characteristics.ope

Ultra-soft and highly stretchable tissue-adhesive hydrogel based multifunctional implantable sensor for monitoring of overactive bladder

A highly stretchable and tissue-adhesive multifunctional sensor based on structurally engineered islets embedded in ultra-soft hydrogel is reported for monitoring of bladder activity in overactive bladder (OAB) induced rat and anesthetized pig. The use of hydrogel yielded a much lower sensor modulus (1 kPa) compared to that of the bladder (300 kPa), while the strong adhesiveness of the hydrogel (adhesive strength: 260.86 N/m) allowed firm attachment onto the bladder. The change in resistance of printed liquid metal particle thin-film lines under strain were used to detect bladder inflation and deflation; due to the high stretchability and reliability of the lines, surface strains of 200% could be measured repeatedly. Au electrodes coated with Platinum black were used to detect electromyography (EMG). These electrodes were placed on structurally engineered rigid islets so that no interfacial fracture occurs under high strains associated with bladder expansion. On the OAB induced rat, stronger signals (change in resistance and EMG root-mean-square) were detected near intra-bladder pressure maxima, thus showing correlation to bladder activity. Moreover, using robot-assisted laparoscopic surgery, the sensor was placed onto the bladder of an anesthetized pig. Under voiding and filling, bladder strain and EMG were once again monitored. These results confirm that our proposed sensor is a highly feasible, clinically relevant implantable device for continuous monitoring OAB for diagnosis and treatment. © 2023 The Authorsope

Clinical validation of serum endocan (ESM-1) as a potential biomarker in patients with renal cell carcinoma

To determine the suitability of serum endocan (ESM-1) levels for diagnosing and monitoring renal cell carcinoma (RCC), we measure serum ESM-1 levels in 56 RCC patients who had undergone radical or partial nephrectomies and 56 age- and sex-matched healthy kidney donors. Measurements were made before and 1 month and 3 months after surgery. The areas under the curve (AUCs) were determined from receiver operating characteristic (ROC) analyses. RCC patients had higher mean serum ESM-1 levels than control subjects (0.59 ± 0.07 vs. 0.52 ± 0.08 ng/mL, P 50 years) individuals (P < 0.001). Among the study cohort, the AUCs for ESM-1 were 0.813 in individuals 50 years of age or younger (n = 55) and 0.637 in individuals older than 50 years (n = 57). In RCC patients, serum ESM-1 levels were reduced 1 month (P = 0.047) and 3 months (P = 0.009) after surgery. These results suggest serum ESM-1 can serve as a serologic biomarker for diagnosing and monitoring RCC, particularly in patients younger than 50 years.ope

Synergic Effect of Metformin and Everolimus on Mitochondrial Dynamics of Renal Cell Carcinoma

Renal cell carcinoma (RCC) frequently recurs or metastasizes after surgical resection. Everolimus, an mTOR inhibitor, is used as a second-line treatment, but the response of RCC to everolimus is insufficient. Metformin is an antidiabetic drug; recent reports have indicated its anti-cancer effects in various cancers, and it is known to have synergistic effects with other drugs. We investigated the possibility of coadministering everolimus and metformin as an effective treatment for RCC. RCC cells treated with a combination of the two drugs showed significantly inhibited cell viability, cell migration, and invasion, and increased apoptosis compared to those treated with each drug alone. An anti-cancer synergistic effect was also confirmed in the xenograft model. Transcriptome analysis for identifying the underlying mechanism of the combined treatment showed the downregulation of mitochondrial fusion genes and upregulation of mitochondrial fission genes by the combination treatment. Changes in mitochondrial dynamics following the combination treatment were observed using LysoTracker, LysoSensor, and JC-1 staining. In conclusion, the combination of everolimus and metformin inhibited RCC growth by disrupting mitochondrial dynamics. Therefore, we suggest that a treatment combining metformin and everolimus disrupts mitochondrial dynamics in RCC, and may be a novel strategy for RCC treatment.ope

Comparison of the Intraparenchymal Biocompatibility of Oxidized Regenerated Cellulose and Porcine Small Intestine Submucosa in Rat Kidney.

PURPOSE: The objective of this study was to evaluate the biological properties of a variety of materials that could be used in partial nephrectomy. MATERIALS AND METHODS: 54 Sprague-Dawley rats(9 for each group) divided into an experimental period of 4 and 12 weeks, respectively. 2 other groups were the negative control for this experiment. We inserted oxidized regenerated cellulose and porcine small intestine submucosa into the defected renal parenchyme in the experimental groups. At the end of the observation periods, the animals were killed and their specimens prepared for histological examination to evaluate the different materials biocompatibility. RESULTS: The reaction of the tissue to the materials diminished with time. After analyzing both periods, the inflammatory reactions to oxidized regenerated cellulose and porcine small intestine submucosa were considered slight. There were no significant differences between oxidized regenerated cellulose and porcine small intestine submucosa. Oxidized regenerated cellulose and porcine small intestine submucosa showed biocompatibility in this test model at both time periods. CONCLUSIONS: It was concluded that there was no difference of biocompatibility between oxidized regenerated cellulose and porcine small intestine submucosa. Both materials are safe, biocompatible materials that can be inserted into the renal parenchyma.ope

Mitochondrial metabolic reprogramming by SIRT3 regulation ameliorates drug resistance in renal cell carcinoma

Clear cell renal cell carcinoma (ccRCC) alters metabolic signals frequently, leading to mitochondrial dysfunction, such as increase of glycolysis and accumulation of lipid. Sirtuin3 (SIRT3) is a key factor for the regulation of both mitochondrial integrity and function. SIRT3 is downregulated and contributes in both cancer development and progression in ccRCC. The aim of this study is to investigate SIRT3-regulated mitochondrial biogenesis in ccRCC. SIRT3 overexpression alone reduced glucose uptake rate and enhanced membrane potential in mitochondria. ccRCC with overexpressed SIRT3 further improved the lethal effects when combined with anticancer drugs (Resveratrol, Everolimus and Temsirolimus). Cell viability was markedly decreased in a dose-dependent manner when treated with resveratrol or mTOR inhibitors in SIRT3 overexpressing ccRCC. In conclusion, SIRT3 improved mitochondrial functions in ccRCC through metabolic reprogramming. Mitochondrial reprogramming by SIRT3 regulation improves the sensitivity to anticancer drugs. The combination of SIRT3 and resveratrol functioned synergistically lethal effect in ccRCC.ope

The Incidence and Clinicopathologic Predictable Factors for Multifocality in the Patients Who had Radical Nephrectomy Performed for Renal Cell Carcinoma

Purpose: We aimed to assess the influence of the clinical and pathological factors on the multifocality of renal cell carcinoma. Materials and Methods: We retrospectively reviewed the records of 467 patients (mean age: 54.6 years) who underwent operation for renal cell carcinoma at this hospital between June 1995 and April 2004. We evaluated the pathologic stage, grade, cell type, the histological pattern, tumor necrosis, the tumor size and the features of multifocality. To determine the clinical significant factors related with multifocality in renal cell carcinoma, we used the chi square test and multiple logistic regression analysis. Results: Of the 467 renal cell carcinomas, 27 cases (5.8%) had multifocal malignant nodules. The average size of the multifocal renal lesions was 17×12×9 mm. The incidence of multifocality was statistically higher in the patients with stage pT3 disease than in those patients with stage pT1 or pT2 disease (p<0.001), and it was also higher for those patients with nuclear grade 3 or 4 than for those patients with nuclear grade 1 and 2 (p=0.005). Only the stage had significant correlation with multifocal disease when performing multiple logistic regression analysis. Conclusions: Our results suggest that the primary tumor stage is a risk factor for multifocality in the patients with renal cell carcinoma. Therefore, more precise preoperative staging of the primary tumor is required in case of performing nephron sparing surgery.ope

Complication rates of the 720 video-assisted minilaparotomy living donor nephrectomies: supplementing clavien classification.

PURPOSE: Laparoscopic living donor nephrectomy (LLDN) has been reported to be as safe and effective as open surgery. We systematically evaluated the safety of video-assisted minilaparotomy surgery-living donor nephrectomy (VAMS-LDN) with use of the modified Clavien classification. MATERIALS AND METHODS: We retrospectively analyzed complications in 720 cases of VAMS-LDN conducted in our institute from 2003 to 2010 by use of the modified Clavien classification of surgical complications. RESULTS: The mean age of the donors was 39.3 years (range, 16 to 66 years) and their mean body mass index was 23.3 kg/m(2) (range, 15.8 to 36.4 kg/m(2)). A total of 67 complications occurred (9.3%). Based on the modified Clavien classification, grade 1, 2a, and 2b complications occurred in 49 (6.8%), 16 (2.2%), and 2 (0.3%) of the donors, respectively. Most grade 1 complications involved mild vascular injuries that were immediately repaired with polypropylene sutures during the surgery. These did not cause any postoperative problems. The other grade 1 complications were wound dehiscence, not requiring secondary closure, and wound site pain in 11 (1.5%) and 5 (0.7%) cases, respectively. Grade 2a complications occurred in 16 (2.2%) cases: 9 (1.3%) involved postoperative transfusions and 1 (0.1%) involved a renal fossa hematoma. One grade 2b complication occurred; it was a lymphocele that resolved with placement of a pigtail catheter. No complications classified as grade 2c or worse occurred. CONCLUSIONS: According to the present analysis of complications, VAMS-LDN is a safe procedure with complication rates comparable to those of LLDN as evaluated in previous studies.ope