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This dose-escalation study evaluated the toxicity and efficacy of different stereotactic body radiation therapy (SBRT) doses for selecting an optimal dose for prostatic adenocarcinoma (PCa). This clinical trial was registered at UMIN (UMIN000014328). Patients with low- or intermediate-risk PCa were equally assigned to 3 SBRT dose levels: 35, 37.5, and 40 Gy per 5 fractions. The primary endpoint was the occurrence rate of late grade ≥2 genitourinary (GU) and gastrointestinal (GI) adverse events at 2 years, while the secondary endpoint was the 2-year biochemical relapse-free (bRF) rate. Adverse events were evaluated using the Common Terminology Criteria for Adverse Events version 4.0. Seventy-five patients (median age, 70 years) were enrolled from March 2014 to January 2018, of whom 10 (15%) and 65 (85%) had low- and intermediate-risk PCa, respectively. The median follow-up time was 48 months. Twelve (16%) patients received neoadjuvant androgen deprivation therapy. The 2-year occurrence rates of grade 2 late GU and GI toxicities were 34 and 7% in all cohorts, respectively (35 Gy: 21 and 4%; 37.5 Gy: 40 and 14%; 40 Gy: 42 and 5%). The occurrence risk of GU toxicities significantly increased with dose escalation (p = 0.0256). Grades 2 and 3 acute GU toxicities were observed in 19 (25%) and 1 (1%), respectively. Grade 2 acute GI toxicity was observed in 8 (11%) patients. No grade ≥3 GI or ≥4 GU acute toxicity or grade ≥3 late toxicity was observed. Clinical recurrence was detected in 2 patients. An SBRT dose of 35 Gy per 5 fractions is less likely to cause adverse events in patients with PCa than 375- and 40-Gy SBRT doses. Higher doses of SBRT should be applied with caution.

The randomized group-greedy method and its customized method for large-scale sensor selection problems are proposed. The randomized greedy sensor selection algorithm is applied straightforwardly to the group-greedy method, and a customized method is also considered. In the customized method, a part of the compressed sensor candidates is selected using the common greedy method or other low-cost methods. This strategy compensates for the deterioration of the solution due to compressed sensor candidates. The proposed methods are implemented based on the D- and E-optimal design of experiments, and numerical experiments are conducted using randomly generated sensor candidate matrices with potential sensor locations of 10,000--1,000,000. The proposed method can provide better optimization results than those obtained by the original group-greedy method when a similar computational cost is spent as for the original group-greedy method. This is because the group size for the group-greedy method can be increased as a result of the compressed sensor candidates by the randomized algorithm. Similar results were also obtained in the real dataset. The proposed method is effective for the E-optimality criterion, in which the objective function that the optimization by the common greedy method is difficult due to the absence of submodularity of the objective function. The idea of the present method can improve the performance of all optimizations using a greedy algorithm.

The ``big'' seismic data not only acquired by seismometers but also acquired by vibrometers installed in buildings and infrastructure and accelerometers installed in smartphones will be certainly utilized for seismic research in the near future. Since it is impractical to utilize all the seismic big data in terms of the computational cost, methods which can select observation sites depending on the purpose are indispensable. We propose an observation site selection method for the accurate reconstruction of the seismic wavefield by process-driven approaches. The proposed method selects observation sites suitable for accurately estimating physical model parameters such as subsurface structures and source information to be input into a numerical simulation of the seismic wavefield. The seismic wavefield is reconstructed by the numerical simulation using the parameters estimated based on the observed signals at only observation sites selected by the proposed method. The observation site selection in the proposed method is based on the sensitivity of each observation site candidate to the physical model parameters; the matrix corresponding to the sensitivity is constructed by approximately calculating the derivatives based on the simulations, and then, observation sites are selected by evaluating the quantity of the sensitivity matrix based on the D-optimality criterion proposed in the optimal design of experiments. In the present study, physical knowledge on the sensitivity to the parameters such as seismic velocity, layer thickness, and hypocenter location was obtained by investigating the characteristics of the sensitivity matrix. Furthermore, the effectiveness of the proposed method was shown by verifying the accuracy of seismic wavefield reconstruction using the observation sites selected by the proposed method., Comment: Accepted manuscript for publication in Geophysical Journal International

Upper urinary tract urothelial carcinoma (UTUC) is a rare disease, often discovered at an advanced stage at diagnosis. Nectin-4 is expressed in a broad range of patients with UTUC and is associated with poor progression-free survival. The receptors of the erythroblastosis oncogene B (ErbB) family are potential therapeutic targets for urothelial carcinoma. Herein, we aimed to investigate the relationship of nectin-4 and ErbB family receptors, namely epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 (HER2) in patients with UTUC. Targeted therapies for these receptors could be used in sequence or in combination for increasing treatment efficiency.We performed immunohisto-chemical analysis for HER2, EGFR, and nectin-4 using tissue microarrays. A total of 98 UTUC patients were included in the study. We investigated the impact of EGFR and HER2 expression status on recurrence-free survival (RFS) and cancer-specific survival (CSS) of all patients.The percentages of patients positive for HER2, EGFR, and nectin-4 were 97%, 70%, and 65%, respectively. The co-expression rates of HER2-EGFR, HER2-nectin-4, and EGFR-nectin-4 were 69%, 64%, and 47%, respectively. The number of patients positive for all three receptors was 47%. Higher HER2 levels were significantly associated with worse CSS and RFS. Higher EGFR levels were associated with a worse CSS.HER2, EGFR, and nectin-4 were highly expressed in UTUC. Combination of HER2-, EGFR-, and nectin-4-targeted therapy may be an effective option for the treatment of patients with UTUC.

To clarify the trends in radical cystectomy and to compare surgical outcomes among surgical approaches focusing on robot-assisted radical cystectomy based on a Japanese nationwide database.The Diagnosis Procedure Combination database was used to extract data on radical cystectomy cases. Trends in open radical cystectomy, laparoscopic radical cystectomy, minimum incision endoscopic radical cystectomy, and robot-assisted radical cystectomy between April 2012 and March 2021 were evaluated. Basic characteristics and peri-operative indicators were compared among the four groups. Propensity score matching was applied to assess the differences between open radical cystectomy and robot-assisted radical cystectomy.During the study period, a decreasing number of open radical cystectomies and an increasing number of minimally invasive radical cystectomies were shown in the total cohort of 28 345 cases. The number of robot-assisted radical cystectomies rapidly increased after government approval in 2018. Minimally invasive radical cystectomies, including robot-assisted radical cystectomies, had a significantly lower complication rate, a shorter length of stay, and a lower blood transfusion rate, but a longer anesthesia time than open radical cystectomies. In the propensity score matching analysis comparing the surgical outcomes of robot-assisted radical cystectomy and open radical cystectomy, similar results were demonstrated, and blood transfusion rates were equivalent.For the past decade, the number of minimally invasive radical cystectomies has steadily increased without compromised surgical outcomes, except for anesthesia time. Robot-assisted radical cystectomies in particular rapidly became widespread in Japan after government approval in 2018 and became a standard surgery within the first 3 years.

This study numerically investigated the characteristics of unsteady aerodynamic forces around a pitching airfoil in the transonic flow regime. The unsteady Reynolds-averaged compressible Navier–Stokes (URANS) equations were solved using a Spalart–Allmaras (SA) turbulence model. The pitching NACA64A010 airfoil at mean angles of attack of 0, 2, 4, and 6 deg with the amplitude of 1 deg and reduced frequency of 0.2 was parametrically simulated. The study found that the trend of the unsteady aerodynamic characteristics around the pitching airfoil, particularly that of the unsteady lift force, significantly changes with the mean-angle-of-attack conditions. The shock-induced boundary-layer separation and reattachment during the pitching oscillation have a crucial role in determining the trend of the unsteady aerodynamic forces, such as the phase-delayed or phase-advanced features for the variation in the angle of attack. The boundary-layer separation during the oscillation, observed at a high mean angle of attack of 6 deg, produces the phase-advanced feature of the lift force. Because the phase-advanced feature is associated with a positive damping effect, the result indicates that the boundary-layer separation may serve as a stabilizing effect on the pitching airfoil motion. The boundary-layer reattachment from separation during the airfoil oscillation, observed at an intermediate mean angle of attack of 4 deg, generates an unusual shock wave movement, resulting in a unique lift loop for the variation in the angle of attack. The study also demonstrated a good prediction accuracy of the URANS with the SA model for the unsteady aerodynamics around the pitching airfoil with the shock and shock-induced boundary-layer separation.

SUMMARY Reconstruction of the distribution of ground motion due to an earthquake is one of the key technologies for the prediction of seismic damage to infrastructure. Particularly, the immediate reconstruction of the spatially continuous wavefield is valuable for decision-making of disaster response decisions in the initial phase. For a fast and accurate reconstruction, utilization of prior information is essential. In fluid mechanics, full-state recovery, which recovers the full state from sparse observation using a data-driven model reduced-order model, is actively used. In this study, the framework developed in the field of fluid mechanics is applied to seismic wavefield reconstruction. A seismic wavefield reconstruction framework based on compressed sensing using the data-driven reduced-order model (ROM) is proposed and its characteristics are investigated through numerical experiments. The data-driven ROM is generated from the data set of the wavefield using the singular value decomposition. The spatially continuous seismic wavefield is reconstructed from the sparse and discrete observation and the data-driven ROM. The observation sites used for reconstruction are effectively selected by the sensor optimization method for linear inverse problems based on a greedy algorithm. The proposed framework was applied to simulation data of theoretical waveform with the subsurface structure of the horizontally stratified three layers. The validity of the proposed method was confirmed by the reconstruction based on the noise-free observation. Since the ROM of the wavefield is used as prior information, the reconstruction error is reduced to an approximately lower error bound of the present framework, even though the number of sensors used for reconstruction is limited and randomly selected. In addition, the reconstruction error obtained by the proposed framework is much smaller than that obtained by the Gaussian process regression. For the numerical experiment with noise-contaminated observation, the reconstructed wavefield is degraded due to the observation noise, but the reconstruction error obtained by the present framework with all available observation sites is close to a lower error bound, even though the reconstructed wavefield using the Gaussian process regression is fully collapsed. Although the reconstruction error is larger than that obtained using all observation sites, the number of observation sites used for reconstruction can be reduced while minimizing the deterioration and scatter of the reconstructed data by combining it with the sensor optimization method. Hence, a better and more stable reconstruction of the wavefield than randomly selected observation sites can be realized, even if the reconstruction is carried out with a smaller number of observations with observation noise, by combining it with the sensor optimization method.

Metastatic castration-resistant prostate cancer (mCRPC) is a genetically heterogeneous disease with a poor prognosis. The prevalence of mutations in homologous recombination repair (HRR) pathway genes, including BRCA1/2, as well as treatment patterns and clinical outcomes, are not well characterized among Japanese men with mCRPC.This multicenter, noninterventional cohort study enrolled Japanese men with mCRPC from 24 institutions between 2014 and 2018. Mutations in the 15 HRR-related genes were assessed using archival primary or metastatic tumor samples. Patterns of sequential therapies for mCRPC were investigated. Patients were followed up for survival evaluation including prostate-specific antigen progression-free survival (PSA-PFS) and overall survival (OS).Of the 143 patients analyzed, HRR-related mutations were detected in 51 patients (35.7%). The most frequently mutated genes were CDK12 (N = 19, 13.3%), followed by BRCA2 (N = 18, 12.6%), ATM (N = 8, 5.6%), and CHEK2 (N = 3, 2.1%). The most common type of first-line therapy for mCRPC was next-generation hormonal agents (NHA, 44.4%), followed by first-generation antiandrogens (FGA, 30.3%), and taxanes (22.5%). Commonly prescribed first-/second-line sequential regimens included FGA/NHA (17.6%), NHA/NHA (15.5%), and NHA/taxanes (14.1%). The median PSA-PFS and OS for the entire cohort were 5.6 and 26.1 months, respectively. Patients carrying BRCA1/2 mutations had numerically shorter PSA-PFS (median 3.3 vs. 5.9 months) and OS (median 20.7 vs. 27.3 months) than those without mutations.In conclusion, approximately one-third of Japanese patients with mCRPC carried mutations in HRR-related genes in this study. The real-world outcomes of mCRPC are poor with conventional therapy, warranting an expansion of treatment options based on genetic abnormalities of the disease.

Lumens, liquid-filled cavities surrounded by polarized tissue cells, are elementary units involved in the morphogenesis of organs. Theoretical modeling and computations, which can integrate various factors involved in biophysics of morphogenesis of cell assembly and lumens, may play significant roles to elucidate the mechanisms in formation of such complex tissue with lumens. However, up to present, it has not been documented well what computational approaches or frameworks can be applied for this purpose and how we can choose the appropriate approach for each problem. In this review, we report some typical lumen morphologies and basic mechanisms for the development of lumens, focusing on three keywords - mechanics, hydraulics and geometry - while outlining pros and cons of the current main computational strategies. We also describe brief guidance of readouts, i.e., what we should measure in experiments to make the comparison with the model's assumptions and predictions.

Approximately, 90% of men with advanced prostate cancer will develop bone metastasis. However, there have been few reports about noninvasive biomarker to detect and predict clinical outcome of bone metastasis (BM) in prostate cancer patients.We examined 1127 patients who underwent prostate biopsy from August 2012 to June 2017. We also investigated bone turnover markers such as bone-specific alkaline phosphatase, type I collagen cross-linked N-terminal telopeptide, C-terminal pyridinoline cross-linked telopeptide of type I collagen, and tartrate-resistant acid phosphatase type 5b (TRACP 5b).A total of 282 patients were diagnosed as prostate cancer with complete clinical data, and 34 patients with bone metastasis. Multivariate analysis revealed C-terminal pyridinoline cross-linked telopeptide of type I collagen, tartrate-resistant acid phosphatase type 5b, and prostate-specific antigen (PSA) were independent biomarkers in detection of BM (p 0.05, respectively). Furthermore, we developed predictive model formula based on tartrate-resistant acid phosphatase type 5b and PSA, for which the area under the curve was 0.95. In patients with bone metastasis, multivariate cox proportional hazards analysis revealed that this model was significantly associated with poor clinical outcome of cancer-specific survival (p 0.05). In validation cohort with 137 patients, we also confirmed the utility of this model for diagnosis of BM (the area under the curve = 0.95).Our developed formula of tartrate-resistant acid phosphatase type 5b in accordance with PSA may serve as the useful tool in diagnosis and prediction of clinical outcome for prostate cancer with bone metastasis.

A material testing apparatus for measuring the biaxial deformation behavior of a polymer tube has been developed to quantitatively evaluate the deformation behavior of polymeric materials. The testing apparatus can apply axial force and internal pressure to a tubular specimen. A noncontact strain measurement system was also developed, and the biaxial strain components and the radius of curvature in the axial direction of the bulging specimen are continuously measured to control the stress path applied to the specimen. Polyethylene tube with an outer diameter of 17 mm and a thickness of 2 mm are used as a test sample. The tubular specimens were subjected to linear stress paths with stress ratios of σΦ:σθ =1:0, 4:1, 2:1, 4:3, 1:1, 3:4, 1:2, 1:4, and 0:1, where σΦ and σθ are the axial and circumferential stress components, respectively, applied to the central area of the bulging specimen. Loading and unloading tests were performed to determine the biaxial true stress-logarithmic plastic strain curves. The strain rate was 1×10-3s-1. From these test results, contours of plastic work and the directions of the plastic strain rates were measured to identify a proper material model for the test sample using the Yld2000-2d yield function (Barlat et al., 2003).

In metastatic renal-cell carcinoma (mRCC), recent clinical trials have shown efficacy of first-line combination therapy, as evidenced by better clinical outcome over target therapy. However, there are insufficient real-world evidences in mRCC patients in Japan.We performed a multicenter retrospective study of 72 mRCC patients who received nivolumab plus ipilimumab as first-line treatment between September 2018 and July 2021. Patient's characteristics, clinical outcomes and safety were retrospectively reviewed. We analyzed overall response rate (ORR), disease control rate (DCR), progression-free survival (PFS), and overall survival (OS) in patients treated with combination therapy.Of all patients, the median age was 70 years (range, 36-86) and the major type of histology was clear cell RCC (n = 55; 76.4%). Progressive disease (n = 25; 34.8%) and irAEs (n = 22; 30.6%) were the most common causes for discontinuing treatment. Median PFS and OS seemed similar between patients who discontinued treatment because of irAEs and for patients who did not (p = 0.360 and p = 0.069, respectively). Importantly, for patients with synchronous metastatic disease at diagnosis (n = 56), nephrectomy before initiating nivolumab plus ipilimumab had a significantly positive impact on better OS when compared to that in patients without nephrectomy (p = 0.028).This study confirms efficacy and safety of nivolumab plus ipilimumab for mRCC patients in real-world settings. Furthermore, nivolumab plus ipilimumab was associated with a better outcome in patients who had undergone nephrectomy at diagnosis for synchronous mRCC.

BACKGROUND: Urinary extracellular vesicles (uEVs) secreted from bladder cancer contain cancer-specific proteins that are potential diagnostic biomarkers. We identified and evaluated a uEV-based protein biomarker for bladder cancer diagnosis and analysed its functions. METHODS: Biomarker candidates, selected by shotgun proteomics, were validated using targeted proteomics of uEVs obtained from 49 patients with and 48 individuals without bladder cancer, including patients with non-malignant haematuria. We developed an enzyme-linked immunosorbent assay (ELISA) for quantifying the uEV protein biomarker without ultracentrifugation and evaluated urine samples from 36 patients with and 36 patients without bladder cancer. RESULTS: Thirteen membrane proteins were significantly upregulated in the uEVs from patients with bladder cancer in shotgun proteomics. Among them, eight proteins were validated by target proteomics, and Ephrin type-A receptor 2 (EphA2) was the only protein significantly upregulated in the uEVs of patients with bladder cancer, compared with that of patients with non-malignant haematuria. The EV-EphA2-CD9 ELISA demonstrated good diagnostic performance (sensitivity: 61.1%, specificity: 97.2%). We showed that EphA2 promotes proliferation, invasion and migration and EV-EphA2 promotes the invasion and migration of bladder cancer cells. CONCLUSIONS: We established EV-EphA2-CD9 ELISA for uEV-EphA2 detection for the non-invasive early clinical diagnosis of bladder cancer. [Image: see text]

The hippocampus and entorhinal cortex are deeply involved in learning and memory. However, little is known how ongoing events are processed in the hippocampal-entorhinal circuit. By recording from head-fixed rats during action-reward learning, here we show that the action and reward events are represented differently in the hippocampal CA1 region and lateral entorhinal cortex (LEC). Although diverse task-related activities developed after learning in both CA1 and LEC, phasic activities related to action and reward events differed in the timing of behavioral event representation. CA1 represented action and reward events almost instantaneously, whereas the superficial and deep layers of the LEC showed a delayed representation of the same events. Interestingly, we also found that ramping activity towards spontaneous action was correlated with waiting time in both regions and exceeded that in the motor cortex. Such functional activities observed in the entorhinal-hippocampal circuits may play a crucial role for animals in utilizing ongoing information to dynamically optimize their behaviors.

Understanding the spatial structure of a city is essential for formulating a spatial strategy for that city. In this study, we propose a method for analyzing the functional spatial structure of cities based on satellite remote sensing data. In this method, we first assume that urban functions consist of residential and central functions, and that these functions are measured by trip attraction by purpose. Next, we develop a model to explain trip attraction using remote sensing data, and estimate trip attraction on a grid basis. Using the estimated trip attraction, we created a contour tree to identify the spatial extent of the city and the hierarchical structure of the central functions of the city. As a result of applying this method to the Tokyo metropolitan area, we found that (1) our method reproduced 84% of urban areas and 94% of non-urban areas defined by the government, (2) our method extracted 848 urban centers, and their size distribution followed a Pareto distribution, and (3) the top-ranking urban centers were consistent with the districts defined in the master plans for the metropolitan area. Based on the results, we discussed the applicability of our method to urban structure analysis.

Background Nivolumab and ipilimumab combination therapy is approved in Japan for unresectable or metastatic renal cell carcinoma. Because the clinical trials supporting the approval of nivolumab and ipilimumab combination therapy included relatively few Japanese patients, post-marketing surveillance was implemented to collate further safety data for nivolumab and ipilimumab combination therapy. Methods Patients with unresectable or metastatic renal cell carcinoma who started nivolumab and ipilimumab combination therapy between September 2018 and December 2019 were registered in this post-marketing surveillance. The observation period was 13 weeks. Safety data included treatment-related adverse events with a particular emphasis on the gastrointestinal-related (colitis, enteritis, diarrhoea and gastrointestinal perforation) and liver-related (hepatic failure, hepatic function abnormal, hepatitis and cholangitis sclerosing) treatment-related adverse events that are listed in the risk management plan for nivolumab and ipilimumab combination therapy. Results Of the 203 patients registered, safety data were available for 159 (119 males/40 females) with a median age of 67 years (range 22–88). Seventy-one patients received nivolumab and ipilimumab combination therapy four times per usual clinical therapy, and 33 continued nivolumab monotherapy thereafter. Any-grade treatment-related adverse events were reported in 102 (64.2%) patients and grade ≥ 3 in 63 (39.6%). Hepatic function abnormalities (13.2%), rash (8.8%) and interstitial lung disease (7.5%) were the most common treatment-related adverse events. Five patients died following treatment-related adverse events. Gastrointestinal-related and liver-related treatment-related adverse events occurred in 10 (6.3%; four with grade ≥ 3 treatment-related adverse events) and 27 (17.0%; 19 with grade ≥ 3 treatment-related adverse events) patients, respectively. Conclusions This post-marketing surveillance in patients with unresectable or metastatic renal cell carcinoma revealed a safety profile for nivolumab and ipilimumab combination therapy consistent with CheckMate 214. Furthermore, no new safety concerns were identified including gastrointestinal-related and liver-related treatment-related adverse events.

The stem, consisting of nodes and internodes, is one of the major organs in seed plants. In contrast to other organs, however, processes of stem development remain elusive, especially when nodes and internodes are initiated. By introducing an intron into theCrerecombinase gene, we established a heat-shock inducible clonal analysis system in a single binary vector and applied it to the stem in the flag leaf phytomer of rice. With detailed characterizations of stem development, we show that cell fate acquisition for each domain of the stem occurs stepwise. Cell fates for a single phytomer and the foot (non-elongating domain at the stem base) were established in the shoot apical meristem by one plastochron before the leaf initiation. The fate acquisition for the node occurred just before the leaf initiation, separating cell lineages for leaves and stems. Subsequently, fates for the axillary bud were established in early leaf primordia. Finally, cells committed to the internode emerged from, at most, a few tiers of cells when the stem epidermis was at the 12∼25 celled stage. Thus, the internode is the last part of the stem whose cell fate is established. This study provides a groundwork to unveil underlying molecular mechanisms in stem development and a useful tool for clonal analysis, which can be applied to various species.

Supplementary Figure S4. A, ACHN cells were transfected with TRIM33 siRNA or control siRNA for 24 h and then stimulated with or without TGF-β1 for 48 h. A wound was then formed by scraping and was measured after 24 h. The results are means {plus minus} S.D. of four independent experiments. *p < 0.05 vs. control siRNA without TGF-β1. White bars indicate 200 μm. B, ACHN cells were transfected with TRIM33 siRNA or control siRNA for 24 h and then stimulated with or without TGF-β1 for 48 h. A cell suspension was added to the upper chamber of matrigel-coated transwell membrane inserts; the lower chamber was filled with the medium and cultured for 12 h. Invasive cells were stained and fluorescence was measured. Values are means {plus minus} S.D. of three independent experiments.

Supplementary Figure S3. A, ACHN cells were transfected with TRIM33 siRNA for 72 h, and then the protein samples were collected and subjected to western blot analysis with anti-TRIM33 antibody. The membrane was re-probed with the anti-β-tubulin antibody. Relative TRIM33 expression is shown as means {plus minus} S.D. of four independent experiments. **p < 0.01 vs. control inhibitor. B, ACHN cells were co-transfected with the reporter construct containing the Smad-binding elements and TRIM33 siRNA or control siRNA, and then were stimulated with TGF-β1 for 24 h. Values are means {plus minus} S.D. of six independent experiments. **p < 0.01; ***p < 0.001 vs. control siRNA.

Supplementary Figure S2. The Caki-2 cells were transfected with TRIM33 siRNA for 72 h, and then protein samples were collected and subjected to western blot analysis by using anti-TRIM33 antibody. The membrane was re-probed with the anti-β-tubulin antibody. Relative TRIM33 expression is shown as means {plus minus} S.D. of four independent experiments. **p < 0.01 vs. control siRNA.

Supplementary Figure S1. A, ACHN cells were transfected with miR-629 mimic or negative control mimic for 72 h. TRIM33 mRNA levels were examined by quantitative real-time PCR. Data are the relative expression normalized to GAPDH mRNA. Values are means {plus minus} S.D. of 3 independent experiments. **p < 0.01 vs. control mimic. B, The expression of miR-629 and TRIM33 mRNAs expression (fold-change: tumor vs. normal) were examined in 41 matched-pair samples of ccRCC by quantitative real-time PCR, and the relationship was evaluated.

Posttransplant malignancy (PTM) is a limiting factor both for patient and allograft survival in kidney transplant recipients (KTRs). We hypothesized that active vitamin D compounds (AVD) could reduce PTM development in KTRs. Ambulatory KTRs in a Japanese prospective cohort were followed from August 2007 to November 2010. The outcome of interest was newly diagnosed PTM. A propensity score (PS) of having received AVDs was estimated using 26 clinically relevant factors. We used the Cox proportional hazards model with stratification by PS tertiles on the assumption that baseline hazard functions differ among tertiles. As sensitivity analyses, we used inverse probability weighting and PS matching. Among 218 participants, the median age was 50 (interquartile range [IQR], 40 to 59) years, 63.3% were male, median time since transplantation was 11.2 (IQR, 5.2 to 17.1) years, and mean estimated GFR was 41.3 (SD, 15.6) mL/min per 1.73 m2. At baseline, 42.2% had been treated with AVDs mainly for glucocorticoid-induced osteoporosis. AVDs used were calcitriol (58.7%) and alfacalcidol (41.3%). During follow-up, PTM developed in 5.4% of 92 AVD users and 8.7% of 126 nonusers. Poor vitamin D status was common in the participants, but the serum 25-hydroxyvitamin D level was not significantly associated with PTM in Cox regression analysis. After stratifying patients by PS tertiles, we found that AVDs were significantly associated with a lower risk of PTM (HR 0.25 [0.07 to 0.82]). Sensitivity analyses yielded similar results. AVDs are potential chemopreventive agents against PTM in KTRs. Cancer Prev Res; 5(10); 1229–35. ©2012 AACR.

Renal cell carcinoma (RCC) is the most common neoplasm of the adult kidney, and clear cell RCC (ccRCC) represents its most common histological subtype. To identify a therapeutic target for ccRCC, miRNA expression signatures from ccRCC clinical specimens were analyzed. miRNA microarray and real-time PCR analyses revealed that miR-629 expression was significantly upregulated in human ccRCC compared with adjacent noncancerous renal tissue. Functional inhibition of miR-629 by a hairpin miRNA inhibitor suppressed ccRCC cell motility and invasion. Mechanistically, miR-629 directly targeted tripartite motif-containing 33 (TRIM33), which inhibits the TGFβ/Smad signaling pathway. In clinical ccRCC specimens, downregulation of TRIM33 was observed with the association of both pathologic stages and grades. The miR-629 inhibitor significantly suppressed TGFβ-induced Smad activation by upregulating TRIM33 expression and subsequently inhibited the association of Smad2/3 and Smad4. Moreover, a miR-629 mimic enhanced the effect of TGFβ on the expression of epithelial–mesenchymal transition–related factors as well as on the motility and invasion in ccRCC cells. These findings identify miR-629 as a potent regulator of the TGFβ/Smad signaling pathway via TRIM33 in ccRCC.Implications: This study suggests that miR-629 has biomarker potential through its ability to regulate TGFβ/Smad signaling and accelerate ccRCC cell motility and invasion. Mol Cancer Res; 13(3); 565–74. ©2014 AACR.

PDF file - 137K, Comparison of baseline characteristics between AVD users and non-users within the tertiles of propensity scores

Supplementary Figure S5. A, Caki-2 cells were transfected with the miR-629 inhibitor or control miRNA inhibitor for 72 h, and then protein samples were collected and subjected to western blot analysis with anti-HNF4α antibody. The membrane was re-probed with the anti-β-tubulin antibody. The relative HNF4α expression is shown as mean {plus minus} S.D. of three independent experiments. B, Protein samples from ccRCC cell lines were collected and subjected to western blot analysis for HNF4α. The relative HNF4α expression is shown as means {plus minus} S.D. of three independent experiments. (C, D) The expression of HNF4α in ccRCC specimens was examined by immunohistochemical staining. One normal kidney tissue and 17 ccRCC samples were examined.