Atsuro Sawada

MK2206 potentiates cisplatin-induced cytotoxicity and apoptosis through an interaction of inactivated Akt signaling pathway

OBJECTIVES To improve conventional chemotherapeutic efficacy, it is important to detect new molecular markers for chemosensitivity and possible accelerating cell-killing mechanisms. In this study, we investigated how MK2206, an allosteric Akt inhibitor, enhances the cisplatin (CDDP)-induced cytotoxicity and apoptosis in urothelial cancer cells. MATERIALS AND METHODS We examined bladder cancer cell lines for the expression of phospho(p)-Akt and its downstream targets by Western blot. The potential antitumor effects were analyzed by MTT assay in vitro and by subcutaneous xenograft models in vivo. The cell invasion was examined by transwell invasion assay, and the activities of the Akt signaling pathway and expression of apoptosis-related proteins were measured by Western blot. RESULTS The expression of p-Akt and its downstream targets was increased in invasive bladder cancer cell lines vs. in noninvasive bladder cancer cell lines. MK2206 (500 nM) inhibited cell invasion in UMUC3 cell line and significantly increased the susceptibility of bladder cancer cell lines to CDDP. When used in combination with CDDP, MK2206 (500 nM) enhanced CDDP-induced cytotoxicity and apoptosis, with suppressed expression of p-Akt and its downstream targets. In vivo MK2206 combined with CDDP effectively suppressed tumor growth in subcutaneous xenograft models. CONCLUSIONS These results suggest that concomitant use of MK2206 could promote the CDDP-induced cytotoxicity and apoptosis in urothelial cancer cell lines through the inhibited expression of the Akt pathway. This combined treatment may provide a new therapeutic option to enhance chemosensitivity in bladder cancer.

Augmented Reality Navigation System for Robot-Assisted Laparoscopic Partial Nephrectomy

We have developed a surgical navigation system for robot-assisted laparoscopic partial nephrectomy (RALPN). In this system, a three-dimensional computer graphics (3DCG) model generated from each patient’s computed tomography image is overlaid on the endoscopic image, and we control it manually to navigate the vascular structure and tumor location in real time. The position and orientation of the 3DCG model is calculated from the optical flow of the endoscopic camera images, which enables the model to move semi-automatically. We conducted 20 navigations for RALPN from April 2014 to December 2016. Our support system worked appropriately and was helpful to localize the tumor and determine the resection line.

Image Overlay Support with 3DCG Organ Model for Robot-Assisted Laparoscopic Partial Nephrectomy

We have developed a surgery support system for robot-assisted laparoscopic partial nephrectomy. In our system, a 3D computer graphics (3DCG) model, which includes a kidney, arteries, veins, and tumors, is saved separately and overlaid on the surgeon’s viewing endoscopic camera image through the operator’s console monitor in real time. The position and orientation of the 3DCG model is calculated from the optical flow of the endoscopic camera images, and the model is moved semi-automatically. The display condition of each part of the 3DCG model is independently controlled anytime by using a mouse, rotary controllers, or key board.

Laparoscopic Forceps with Force Feedback

There are two main aspects of safety and effectiveness in laparoscopic surgery. The first is ensuring an appropriate operation field and maintaining this throughout the procedure. The second is finding the correct tissue plane and applying appropriate traction and counter-traction. To accomplish these requirements, surgeons must know the appropriate pressure to apply to the target organ or tissue. For example, weak operation of the forceps in the left hand during laparoscopic surgery leads to poor visibility owing to a small operational field. Furthermore, poor traction at the point of incision on the dissected plane decreases the dissection efficiency. In contrast, when excessive force is applied, there is an increased risk of organ injury or bleeding from capillary vessels during traction; this is clearly detrimental to the overall safety of the procedure. It is difficult for surgeons to master a feeling for the appropriate pressure to apply. Imitating the techniques of skilled surgeons is essential; however, surgical techniques cannot be imitated on first sight. With the aim of assisting this learning process, we developed Forceps Guiding Correct Operation (FOGCOP), new laparoscopic forceps with sensors. Although they are the same shape as Maryland dissecting forceps, FOGCOP can measure the pressure applied on the shaft of the forceps in three axis directions (X, Y, and Z) and on the jaw. The measured pressures are displayed in real time. Surgeons can insert this device into a 5-mm trocar in the same manner as normal forceps. We conducted experiments to verify the effectiveness of the device. 30 students with no experience of laparoscopic surgery participated in this study. Using a training box, students performed a task to press and pull a rubber plate. We compared the performance of a group of students using FOGCOP (group 1, n = 10) with that of a group using conventional forceps (group 2, n = 10). The results suggest that the feedback provided by FOGCOP may be useful for understanding the force delivered by forceps. To verify the usefulness of the device, FOGCOP was also used in laparoscopic nephrectomy of a pig, a procedure that is part of the training course for laparoscopic surgery. It was possible to dissect a tissue plane in the same manner as with conventional forceps. However, the wire from the sensor sometimes interfered with operation. In future, we intend to upgrade FOGCOP by including a wireless sensor, to improve operability to be closer to that of normal forceps, and to allow this device to be used as an educational tool.

Laparoscopic excision of an acquired ureteral diverticulum: A case report: Laparoscopy for ureteral diverticulum

Ureteral diverticula, especially acquired diverticula, are rare. Surgery is indicated when a diverticulum is accompanied by complications such as stones, pyelonephritis, stenosis, and signs of malignancy. A 59‐year‐old woman visited our urology department with asymptomatic macrohematuria. Enhanced CT scan revealed a right ureteral diverticulum with a 16‐mm diameter that contained two tiny stones inside. After 8 months, the size of these stones increased; therefore, the patient underwent laparoscopic resection of the ureteral diverticulum and end‐to‐end anastomosis of the ureter. Subsequent histopathology of the excised specimen revealed an acquired diverticulum. Follow‐up intravenous pyelography showed adequate urine passage with only minor dilatation of the pelvis at 3 months after the operation. The laparoscopic approach is believed to be feasible for ureteral diverticula cases that require ureteral end‐to‐end anastomosis.

A 45,X/46,XY Male with Orchidopexy Diagnosed with Mixed Germ Cell Tumor After 21-year Follow-up

A case of a 45,X/46,XY boy with gonadal dysgenesis is presented. The patient showed hypospadias and right undescended testis. He underwent underwent repair surgery for hypospadias, right orchidopexy, and bilateral testicular biopsy. Testicular biopsy revealed no malignant finding. He was followed-up annually by scrotum palpation. When the patient grew up to 24 years old, he was diagnosed to have right testicular tumor. High orchiectomy revealed pT1 seminoma. The management of undescended testis in men with gonadal dysgenesis and disordered sexual development is discussed.

MP02-12 INTRAVESICAL PROSTATIC PROTRUSION IS NOT THE SAME IN ITS SHAPE: EVALUATION BY PREOPERATIVE CYSTOSCOPY AND OUTCOME IN HOLEP

INTRODUCTION AND OBJECTIVES: Intravesical prostatic protrusion (IPP) has been known as a predictor of efficacy not only for medical treatment such as alpha 1 blocker and dutasteride, but also for holmium laser enucleation of the prostate (HoLEP). However, the IPP is considered not the same in its shape because middle lobe and/or lateral lobes can protrude into bladder. Here, we evaluated the shape of IPP by cystoscopy and analyzed the outcome. METHODS: We reviewed charts of patients who had undergone HoLEP in Kyoto University Hospital from January 2006 to June 2016. Among 222 cases, 157 cases were evaluable for IPSS, uroflowmetry, IPP and its shape by preoperative flexible cystoscopy in outpatient clinic. IPP was classified into 5 groups: A, no protrusion; B, middle lobe only; C, lateral lobe only; D, bilateral lobe; E, B+C or B+D. Paired match analysis with similar IPP and other parameters was performed between the group with middle lobe protrusion (B+E, n1⁄433) and the one without it (C+D, n1⁄433). RESULTS: Table 1 shows the number of patients, age, score of total IPSS, QOL score, Qmax and IPP in the five groups. The group A (no protrusion) had a significantly higher Qmax than other groups. Groups with middle lobe protrusion (B or E) had a better tendency in changes in total IPSS score and Qmax. Paired match analysis shown in Table 2 demonstrated that the group with middle lobe protrusion had a significantly greater improvement of total IPSS score than the one without it (-16.6 vs. -10.8. p<0.01). Among them with less than 16 mm of IPP, all of patients with middle lobe protrusion improved IPSS, while only 76.5% (13/ 16) of patients without it were improved. CONCLUSIONS: Patients with middle lobe protrusion had a greater improvement of IPSS in HoLEP than those having similar length of IPP without middle lobe protrusion. IPP should be clinically divided into two groups at least.

MP44-19 HIGH-THROUGHPUT CHEMICAL SCREENING FOR SENSITIZATION OF BLADDER CANCER TO GEMCITABINE AND CISPLATIN CHEMOTHERAPY

CTL/Treg ratio in the presence of PD-L1+ TIM (favorable). This indicates that PD-L1+ TIM and PD-L1 margin expression were independently predictive in absence of HPV, the interaction between CTL/Treg ratio and PD-L1+ TIM was predictive irrespectively of HPV-status. CONCLUSIONS: A PD-L1 expression pattern predominantly at the tumor-stroma margin predicts good prognosis, while the negative predictive value of PD-L1+ TIM appear to be compensated by a high CTL/Treg ratio. All independent prognostic factors are PD-L1 related parameters. These results strengthen the rationale for anti-PD-1/PD-L1 immunotherapy in penile carcinoma.

Abstract 5046: High-throughput chemical screening for sensitization of bladder cancer to gemcitabine and cisplatin chemotherapy

Background Gemcitabine and cisplatin chemotherapy (GC) is the current standard regimen for locally advanced and metastatic bladder cancer (BC). Despite a relatively high initial response rate, some cases do not regress (intrinsic resistance) and the remaining cases often show regrowth after initial shrinkage (acquired resistance). To identify novel therapeutic agents for overcoming these resistances, we applied a high-throughput screening of chemicals administered in combination with GC. Methods and Findings As a high-throughput screening, 2100 compounds were administered alone or in combination with GC to human BC cell lines (J82, UMUC-3). Cell viability was determined after 3-day incubation and chemicals that enhanced inhibitory effect of GC were screened. The initial screening identified 26 compounds and further validation narrowed them into the most synergistic agent disulfiram (DSF), an FDA-approved drug for alcoholism. Combination index assay showed synergistic effects of DSF with cisplatin but not with gemcitabine in J82, UMUC-3, T24, HT1197 and HT1376 cells. Co-administration of DSF significantly enhanced apoptosis by GC treatment in UMUC-3 and T24 cells, with significant increase of reactive oxygen species (ROS) production. DSF-induced apoptosis was attenuated in the presence of N-acetyl L-cysteine and Trolox, ROS scavengers. Use of DSF in combination with GC (GCD) significantly inhibited tumor growth of UMUC-3 subcutaneous xenograft on athymic mice (by 39% compared with GC alone, p = 0.02). GCD regimen was as tolerable as GC and no significant differences were observed in body weight of treated mice between the two regimen. Conclusion Repositioning of DSF to a chemotherapy sensitizer is a promising treatment strategy, which can be translated rapidly in the future. Citation Format: Yuki Kita, Takashi Kobayashi, Norihiko Masuda, Atsuro Sawada, Yoshiyuki Matsui, Takahiro Inoue, Tomomi Kamba, Osamu Ogawa. High-throughput chemical screening for sensitization of bladder cancer to gemcitabine and cisplatin chemotherapy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5046.