Galectin-9 expression as a poor prognostic factor in patients with renal cell carcinoma.

Recently, the effectiveness of anti-programmed death 1 (PD-1) antibody therapy in the treatment of renal cell carcinoma (RCC) has been established. Nevertheless, efficacy has been reported to be limited to only 10-30% of patients. To develop more effective immunotherapy for RCC, we analyzed the immunological characteristics in RCC tissues by immunohistochemistry (IHC). We prepared a tissue microarray that consisted of tumor tissue sections (1 mm in diameter) from 83 RCC patients in Kanagawa Cancer Center between 2006 and 2015. IHC analysis was performed with antibodies specific to immune-related (CD8 and Foxp3) and immune checkpoint (programmed death ligand 1 (PD-L1) and 2 (PD-L2), B7-H4 and galectin-9) molecules. The numbers and proportions of positively stained tumor cells or immune cells were determined in each section. From multivariate analysis of all 83 patients, higher galectin-9 expression was detected as a factor associated with worse overall survival (OS) (P = 0.029) and that higher stage and higher B7-H4 expression were associated with worse progression-free survival (PFS) (P < 0.001 and P = 0.021, respectively). Similarly, in multivariate analysis of 69 patients with clear cell RCC, though not statistically significant, there was a trend for association between higher galectin-9 expression and worse OS (P = 0.067), while higher stage was associated with worse PFS (P < 0.001). This study suggests that higher galectin-9 expression is an independent adverse prognostic factor of OS in RCC patients. Therefore, to develop more effective personalized immunotherapy to treat RCC, it may be important to target not only PD-1/PD-L1, but also other immune checkpoint molecules such as galectin-9.

Prognostic value of OCT4A and SPP1C transcript variant co-expression in early-stage lung adenocarcinoma.

BACKGROUND: Octamer-binding transcription factor 4A (OCT4A) is essential for cell pluripotency and reprogramming both in humans and mice. To date, however, the function of human OCT4 in somatic and/or tumour tissues is largely unknown. METHODS: RT-PCR was used to identify full-length splice forms of OCT4 transcripts in normal and cancer cells. A FLAG-tagged OCT4 genomic transgene was used to identify OCT4-positive cancer cells. A potential role for OCT4 in somatic cancer cells was examined by cell ablation of OCT4-positive cells using promoter-driven diphtheria toxin A. OCT4 and secreted phosphoprotein 1 (SPP1) transcripts in early-stage lung adenocarcinoma tumours were analysed and compared with pathohistological features. RESULTS: The results show that, unlike in murine cells, OCT4A and OCT4B variants are transcribed in both human cancer cells and in adult tissues such as lung, kidney, uterus, breast, and eye. We found that OCT4A and SPP1C are co-expressed in highly aggressive human breast, endometrial, and lung adenocarcinoma cell lines, but not in mesothelial tumour cell lines. Ablation of OCT4-positive cells in lung adenocarcinoma cells significantly decreased cell migration and SPP1C mRNA levels. The OCT4A/SPP1C axis was found in primary, early-stage, lung adenocarcinoma tumours. CONCLUSIONS: Co-expression of OCT4 and SPP1 may correlate with cancer aggressiveness, and the OCT4A/SPP1C axis may help identify early-stage high-risk patients with lung adenocarcinoma. Contrary to the case in mice, our data strongly suggest a critical role for OCT4A and SPP1C in the development and progression of human epithelial cancers.

Conclusive Evidence for OCT4 Transcription in Human Cancer Cell Lines: Possible Role of a Small OCT4-Positive Cancer Cell Population.

The role of octamer-binding transcription factor 4 (OCT4) in human cancer is still debated. Although many studies have been published on human OCT4, determining which of the findings are accurate or which are false-positives is currently challenging. We thus developed the most reliable method to date for highly specific and comprehensive detection of genuine OCT4-transcript variants without false-positive results. Our results provided clear evidence that the transcripts of OCT4A, OCT4B, OCT4B1, and other novel splicing variants are indeed present in many cancer cell lines, but are rarely detected in normal tissue-derived differentiated cells. Using the tagged genomic transgene, we then verified endogenous OCT4A translation in cancer cell subpopulations. Moreover, analysis of possible other protein isoforms by enforced expression of OCT4B variants showed that the B164 isoform, designated human OCT4C, is preferentially produced in a cap-dependent manner. We confirmed that the OCT4C isoform, similar to OCT4A, can transform non-tumorigenic fibroblasts in vitro. Finally, ablation of OCT4-positive cells using promoter-driven diphtheria toxin A in high malignant cancer cells caused a significant decrease in migration and Matrigel invasion. These findings strongly suggest a significant contribution of OCT4 to the phenotype of human cancer cells. Stem Cells 2018.

[Myeloid Sarcoma of the Testis].

A 68-year-old man with swelling of his left testis was referred to our hospital. There was no history of hematologic disease. The diameter of the testicular mass was 40 mm. We performed a left orchiectomy. Histopathologic examination revealed diffuse infiltration of immature neoplastic cells with a high nuclearcytoplasmic ratio. Immunohistochemical analysis revealed that cells were positive for myeloperoxidase, while T-cell and B-cell markers were negative. There was no evidence of leukemia cells in peripheral blood or in bone marrow. The tumor was diagnosed as a testicular isolated myeloid sarcoma. Eleven months after the orchiectomy, enlargement of the para-aortic lymph node occurred. There was no evidence of leukemia cells in the peripheral blood or bone marrow. The patient is undergoing chemotherapy for recurrence of myeloid sarcoma. Myeloid sarcoma is an extramedullary tumor composed of immature myeloid cells. Myeloid sarcoma primarily involving the testis is rare. However, it is important to consider it as a differential diagnosis because it has a poor prognosis.

[Pathological Complete Response of Metastatic Testicular Tumor with Persistent Low Level Positive Human Chorionic Gonadotropin after Chemotherapy].

We describe a case of testicular tumor with multiple metastasis to the lung,retoroperitoneal lymph node, and brain. After chemotherapy the retroperitoneal lymph node and brain metastasis disappeared,but the multiple pulmonary metastases but not disappear,although they were reduced in size. Since the human chorionic gonadotoropin (HCG) was persistently dected at a low level,we performed a testosterone tolerance test. The HCG level became undetectable for a while,but was detected at a low level again. Then the patient underwent residual tumor removal of some of the residual pulmonary disease,which was diagnosed as tumor necrosis. The patient has been followed on an ambulatory basis after surgery for 12 months without recurrence. In this case a definitive diagnosis was difficult,because of the low positive level of HCG.

[A Case of Long Survival in Aggressively Growing Small Cell Carcinoma of the Bladder Successfully Treated by Combined Chemotherapy of Methotrexate, Etoposide and Cisplatin].

A 74-year-old man underwent transurethral resection for a bladder tumor (TURBT). The pathological diagnosis was urothelial carcinoma, grade 3 pT2 at least. He desired preservation of the bladder. Thus, MEC (methotrexate 100-150 mg/body (day 1), etoposide 100 mg/m2 (day 2-4), cisplatin 20 mg/m2 (day 2-6)) chemotherapy was administered for 2 courses. The next year, he had a relapse in the bladder, and the pathological diagnosiswasurothelial carcinoma, grade 2 pTa and pTis. He underwent Calmette-Guerin Bacillus (BCG) immunotherapy for 6 courses that resulted in a complete response without recurrence for 6 years. Six months after the latest examination, he complained of difficulty in voiding. An 8 cm tumor in the bladder and enlargement of obturator lymph node were detected. The pathological diagnosis by TURBT was small cell carcinoma. He rejected cystectomy, so we applied MEC therapy again. After 2 courses of MEC therapy, the bladder tumor and lymphadenopathy markedly shrunk in image and almost disappeared subsequently. The patient refused further therapy, but he had been followed without recurrence for 48 monthsafter the chemotherapy.

[Ipsilateral Occurrence of Renal Pelvic Carcinoma after Partial Nephrectomy for Renal Cell Carcinoma].

A 70-year-old man underwent left partial nephrectomy for renal cell carcinoma (pT1aN0M0). One year after the surgery, he presented with hematuria and fatigue. Computed tomography showed a left 8 cm renal tumor and multiple liver and lung metastases. We performed percutaneous renal and liver biopsy with echo guidance. The diagnosis of both kidney and liver was urothelial carcinoma. He died 3 weeks after the diagnosis. Ipsilateral occurrence of the pelvic renal carcinoma after partial nephrectomy for renal cell carcinoma is extremely rare. To our knowledge, this case is the first to be reported in Japan and elsewhere.

[A Case of Metastatic Seminomatous Testicular Tumor with Complicated Diagnosis by FDG-PET].

18F-fluorodeoxy glucose positron emission tomography (FDG-PET) for evaluation of the post chemotherapy residual tumor of the seminomatous testicular germ cell tumor is recommended by several guidelines. We report a case whose residual tumor was evaluated by FDG PET but the results were difficult to interpret. A 41-year-old male with left seminomatous germ cell tumor of the testis and 60 mm retroperitoneal lymph node (RPLN) metastasis was referred to our hospital. The International Germ Cell Consensus Classification (IGCCC) was good prognosis. After high orchiectomy, three cycles of bleomycin, etoposide, and cisplatin (BEP) chemotherapy normalized the tumor marker and the RPLN decreased to 15 mm. The standardized uptake value (SUV) max at the RPLN by FDG-PET was 2.93. Although residual viable cells were suspected, the SUV max was relatively low. Thus surveillance without additional therapy was selected. After observation for 25 weeks, the tumor grew to 25 mm. Then four cycles of paclitaxel, ifosfamide, and cisplatin (TIP) chemotherapy were indicated for the recurrence. The RPLN was decreased to 15 mm, but the SUV max was still as high as 2.67 at 6 weeks after the last chemotherapy. We dissected the residual tumor suspecting viable cancer, but the pathological examination revealed necrotic tissue without any viable cells. He has had no signs of recurrence for 1 year and 9 months after the operation.

Complete Response to Pembrolizumab in a Patient with Castration-Resistant Prostate Cancer with Both BRCA Positivity and a High Frequency of Microsatellite Instability: A Case Report.

Introduction: There have been few reports of patients for whom a cancer gene panel test for solid tumors revealed the simultaneous presence of BRCA mutation and microsatellite instability (MSI)-high status. BRCA mutations have been reported in 13% of castration-resistant prostate cancer (CRPC) patients, and 3.1% of prostate cancer cases are MSI-high/mismatch repair deficient. Case Presentation: A 71-year-old man with a history of urinary retention was referred to our department for clinically suspected prostate cancer and a high prostate-specific antigen (PSA) level (141 ng/mL). MRI revealed features of prostate cancer invading the bladder, seminal vesicles, and rectum. A histopathological examination of a transperineal needle biopsy specimen obtained from the prostate revealed adenocarcinoma. Bone scintigraphy revealed multiple metastases. The patient was treated with abiraterone acetate combined with androgen deprivation therapy followed by local radiation. Rectal wall thickening and lymph node metastasis were also observed, and docetaxel was administered. A cancer gene panel test was positive results for BRCA2 mutation with a MSI-high. After six courses of docetaxel, lymph node enlargement was observed and olaparib was initiated. Two months later, the metastatic lesions showed enlargement and the PSA level increased. Subsequently, pembrolizumab was administered. At 2 to the patient months after the initiation of pembrolizumab administration, PSA levels decreased to <0.025 ng/mL and the rectal lesions and lymph node metastases disappeared. The patient was continuing to receive pembrolizumab without any apparent adverse events or exacerbations, 9 months after initiation. Conclusion: We herein report a case in which pembrolizumab treatment resulted in a complete response in a CRPC patient with both a BRCA2 mutation and an MSI-high status.

Transgenic Xenopus laevis with the ef1-α promoter as an experimental tool for amphibian retinal regeneration study.

Complete retinal regeneration occurs after the removal of the whole tissue in mature Xenopus laevis, as well as in the newt. Here, we produced F1 and F2 lines of transgenic X. laevis containing an EGFP gene under a translation elongation factor 1-α (ef1-α) promoter and investigated how the gene is reactivated in retinal pigmented epithelial (RPE) cells when the neural retina (NR) is removed. The results showed that EGFP expression is reduced in the adult ocular tissues of nonmanipulated transgenic animals, and EGFP-expressing cells are occasionally found heterogeneously in the lens, NR and RPE tissues. During retinal regeneration, the EGFP gene is reactivated in the RPE and ciliary marginal cells. Transgenic animals were also used for a transplant study because of the genetic marker of the donor tissue. Transplanted RPE clearly transdifferentiated to regenerate the retina in the ocular chamber. This study is, to our knowledge, the first report of a transgenic study of amphibian retinal regeneration, and the approach is promising for future molecular analyses.

C-reactive protein in patients with advanced metastatic renal cell carcinoma: usefulness in identifying patients most likely to benefit from initial nephrectomy.

OBJECTIVE: C-reactive protein (CRP) is considered a useful serum marker for patients with RCC. However, its clinical utility in advanced metastatic renal cell carcinoma (AM-RCC), particularly in deciding whether to perform nephrectomy at the onset, is not well studied. PATIENTS AND METHODS: We retrospectively evaluated 181 patients with AM-RCC, including 18 patients underwent potentially curative surgery, 111 underwent cytoreductive nephrectomy, and 52 received medical treatment only. CRP cutoff points were determined by receiver operating characteristic (ROC) curve analysis. Kaplan-Meier and Cox regression analyses were used for survival tests. RESULTS: ROC analysis suggested that grouping patients according to 3 CRP ranges was a rational model. Patients with highly elevated CRP (≥67.0 mg/L) presented remarkably poor prognosis despite treatment (nephrectomy or medical treatment only). Cox regression models demonstrated that risk factors of overall survival for patients who underwent nephrectomy were the CRP ranges defined in this study (≤18.0 mg/L, >18.0 and <67.0 mg/L, and ≥67.0 mg/L), ECOG PS (0, 1, and ≥2), and number of metastatic organ sites (0-1 and ≥2). The retrospective design is a limitation of this study. CONCLUSION: Our study demonstrated that the serum CRP level is a statistically significant prognostic parameter for patients with AM-RCC. The data also indicated that pretreatment serum CRP level provides useful prognostic information that helps in deciding whether to perform initial nephrectomy for patients with AM-RCC.

[Assessment of clinical features in solitary minimum size (<1 cm) bladder cancer].

We retrospectively evaluated primary non-muscle-invasive bladder cancer diagnosed between 1999 and 2008 at 2 facilities (Kawasaki Municipal Hospital and Yokohama Minami Kyosai Hospital). Size (< 1 cm) solitary bladder cancer statistically evaluated the characteristics. Out of 463 bladder cancers, 52 were minimum-size solitary pTa bladder cancers less than 1 cm in diameter. The average follow-up period was 50.9 months. The recurrence rate of the minimum-size bladder cancer was significantly lower than that of bladder cancers of other sizes (1 to 3 cm or ≥ 3 cm). The 3-year non-recurrence rate was 80.7,71.0,and 62.9% in each group (< 1, 1 to 3, ≥ 3 cm). High-grade minimum size bladder cancer (pTa) showed a significantly higher recurrence rate than the low-grade cases (P = 0.0101). Intravesical chemotherapy with anti-cancer drugs significantly reduced the intravesical recurrence rate in the low-grade minimum-size bladder cancer group (P = 0.0418). There was no statistically significant difference in either the average recurrence number or the rate of multiple recurrences between the minimum-size tumor group and the 1 to 3 cm tumor group. Minimum size bladder cancer had a lower recurrence rate than tumors of other sizes; however, there were no differences in other characteristics between the groups. Therefore, sufficient treatment, in accordance with the guidelines, should be administered for minimum size tumors as well as tumors of other sizes.

[Assessments of progression in non-muscle-invasive bladder cancer].

We retrospectively studied 463 patients with primary non-muscle-invasive bladder cancer diagnosed between 1999 and 2008 at two facilities (Kawasaki Municipal Ida Hospital and Yokohama Minami Kyosai Hospital). In this study, disease progression was defined as invasion to the muscle or further (upstage) and presence of metastasis (metastasis). We detected progression in 22 cases, including 18 upstages and 4 metastasis. Univariate analysis showed that factors associated with progression were T category (pT1 p< 0.0001), grade (high grade p< 0.0001, G3 p< 0.0001) and number of tumors (multiple p=0.0213). Multivariate analysis showed that the only equivocal factor associated with progression was T category (T1). Use of a second tansurethral resection for high-grade pT1 cases was unrelated to progression. Among the patients with progression, many had a more advanced T category at the time of radical treatment, and the results of treatment were poor. The factors associated with progression of bladder cancer should be investigated in more detail, so that early radical treatment can be initiated in eligible patients.

[A case of primary seminal vesicle cancer detected by FDG-PET/CT].

We reported a case of primary seminal vesicle cancer, detected by FDG-PET/CT. A 65-year-old man with constipation and appetite loss was admitted to our hospital. An ultrasound examination revealed evidence of bilateral hydronephrosis. He was diagnosed as acute post renal failure, and nephrostomy was done. CT and MRI showed a solid mass in the area of seminal vesicle. He underwent transrectal core biopsy, which histologically showed poorly differentiated adenocarcinoma. Immunohistochemistry showed the tumor to be CA125 positive, CEA positive and CK7 positive but PSA negative. FDG-PET/CT revealed an increased uptake of FDG only in the area of seminal vesicle. Serum CA125 was elevated and PSA stayed within normal limit. Primaly rectal carcinoma was ruled out by colonoscopy. The result of transperineal prostate biopsy was negative. We diagnosed him as suffering from primary seminal vesicle carcinoma. Anti-androgen blockade and radiotherapy to whole pelvis were performed, and serum CA125 level was improved. But, 6 months later serum CA125 re-elevated and 19 months later multiple liver metastases were noted. The patient received two cycles of docetaxel and cisplatin chemotherapy, however he developed pulmonaly embolism and rectal bleeding by tumor invasion and he died of his disease 22 months after the diagnosis.

Characterization of a novel type I keratin gene and generation of transgenic lines with fluorescent reporter genes driven by its promoter/enhancer in Xenopus laevis.

We investigated the characteristics of a novel type I keratin gene in Xenopus laevis during ontogenesis. The transcript was first detected in the posterior region at the late neurula stage, and then restricted to the fin and external gill during embryogenesis. To examine the transcriptional regulation of the keratin gene in vivo, we generated transgenic lines with fluorescent reporter genes driven by its 4.2-kb upstream sequence. The promoter/enhancer activity recapitulated the endogenous gene expression during embryogenesis. Sequential deletion analyses revealed that the regions proximal to the promoter were essential for fin-specific expression. Reporter expression was detected in various organs, including the fin and gill. In particular, robust expression was observed in the developing limbs and gill. The reporter fluorescence rapidly decreased with internal gill resorption during metamorphosis. The transgenic lines carrying the promoter/enhancer should represent valuable tools for elucidating the formation, development and resorption of various organs, especially the gill.

A higher De Ritis ratio (AST/ALT) is a risk factor for progression in high-risk non-muscle invasive bladder cancer.

BACKGROUND: High-risk non-muscle invasive bladder cancer (NMIBC) is thought to be associated with a higher risk of recurrence and progression. A recent study revealed that a high De Ritis ratio was a risk factor in some solid malignancies. This study examined the importance of the De Ritis ratio as a prognostic marker in high-risk NMIBC. MATERIALS AND METHODS: A total of 138 patients who were initially diagnosed with high-risk NMIBC between January 2012 to December 2016 were enrolled in this study. The criteria for the high-risk classification followed the EAU guidelines. The recurrence-free and progression-free survival of the higher and lower De Ritis ratio groups were compared. The cut-off value of the De Ritis ratio was set at 1.35, based on a receiver operator curve analysis. RESULTS: The median observation period was 50.3 months. Among these patients, 32 (23.1%) patients developed recurrent disease and 15 (10.9%) patients showed progression. A multivariate analysis revealed that non-BCG treatment was an independent risk factor for recurrence, and a higher De Ritis ratio was an independent risk factor for cancer progression. CONCLUSIONS: The De Ritis ratio might be a risk factor for progression in high-risk NMIBC.

[Pancreatic metastasis from renal cell carcinoma 25 years after radical nephrectomy].

We report a case of pancreatic metastasis from renal cell carcinoma detected 25 years after radical nephrectomy. A 74-year-old man, who had undergone radical nephrectomy for renal cell carcinoma at age 49, was found by computed tomography to have a strongly enhanced mass on the pancreatic head. The patient underwent pancreaticoduodenectomy and the pathological diagnosis was metastatic renal cell carcinoma. This was evidently a slow growing tumor because the metastatic pancreas tumor was well demarcated and the metastasis was found 25 years after the primary operation. Aggressive surgical treatment of isolated metastatic lesions offers a chance of long-term survival. Patients with a history of RCC should undergo a long-term follow-up to detect and evaluate metastasis to pancreas as well as other organs.

Determinative roles of FGF and Wnt signals in iris-derived lens regeneration in newt eye.

Total regeneration of experimentally excised lens from the dorsal part of the iris-pigmented epithelium of newts has been a key model of tissue regeneration via cells originating from a foreign tissue. Due to the strict spatial restriction of the lens origin in the newt iris, it has often been assumed that only the dorsal iris cells are endowed with an intrinsic potential to give rise to lens tissues. However, our reinvestigation of the process revealed completely different mechanisms underlying lens regeneration and its spatial restriction, comprising the following two steps: (i) Fibroblast growth factor (FGF) 2-dependent proliferation of iris-pigmented epithelium and activation of early lens genes (Pax6, Sox2, MafB) over the entire circumference of the iris; and (ii) dorsal iris-restricted activation of the canonical Wnt signals (involving Wnt2b and Frizzeld4) that leads to localized expression of late lens genes (Prox1, Sox1, beta-crystallin). Injection of FGF2 into normal eyes specifically elicited the second lens development from the dorsal iris, and the administration of recombinant Wnt3a to the cultured iris-pigmented epithelium caused even ventral iris-derived lens development. Thus, it is concluded that the regulation of FGF2 and Wnt signals is a determinative of the iris-derived lens regeneration in the newt eye.

[PRIMARY LOCALIZED AL AMYLOIDOSIS OF THE BLADDER MIMICKING RECURRENCE OF BLADDER CARCINOMA].

Primary bladder amyloidosis is a rare disease, with approximately 200 cases documented in the literature. We herein present a 85-year-old Japanese man who has undergone a transurethral resection of a bladder tumor (TURBT) and has regularly been followed up after surgery. Since cystoscopy revealed mucosal irregularity, he has got a TURBT again for a suspicion of recurrence. There were no malignant findings in pathological diagnosis and we diagnosed as amyloidosis because it showed positive by Congo-red staining. We added immunohistological diagnosis to diagnose as localized AL amyloidosis of the bladder finally.

[RETROPERITONEAL LIPOSARCOMA WITH MULTIPLE RECURRENCE OF LUNG METASTASES TREATED BY MULTIMODAL THERAPY CENTERING ON THE OPERATION: A CASE REPORT].

A 34-year-old man presented with scrotal pain and slight fever. The scrotal pain was improved by the treatment of antibiotics, but the slight fever remained and an abdominal protuberance appeared. Computed tomography showed a 22 cm abdominal tumor with lipid density. He was then referred to our hospital. He was diagnosed as retroperitoneal liposarcoma and a surgical resection was performed for retroperitoneal tumor and surrounding organs. Histopathological diagnosis was dedifferentiated liposarcoma. 3 months after surgery, a PET/CT scan showed multiple lung metastases. We treated the patient with AI therapy by doxorubicin and ifosfamide. After 6 courses were performed, a complete response was achieved. 30months after the initial surgery, a PET/CT scan showed there was just one metastasis which was in the left lung. Thoracoscopic lung tumor resection was performed. Histopathological diagnosis was metastatic dedifferentiated liposarcoma. As adjuvant therapy, we treated with IE therapy by ifosfamide and VP-16. 3 courses were performed. 3 years and 6 months after the first surgery, he has had no recurrence up to the present day.