Mototaka Sato

EMMPRIN Promotes Angiogenesis, Proliferation, Invasion and Resistance to Sunitinib in Renal Cell Carcinoma, and Its Level Predicts Patient Outcome

Purpose Extracellular matrix metalloproteinase inducer (EMMPRIN) has been reported to play crucial roles, including in angiogenesis, in several carcinomas. However, the correlation between EMMPRIN levels and angiogenesis expression profile has not been reported, and the role of EMMPRIN in renal cell carcinoma (RCC) is unclear. In the present study, we evaluated the association of EMMPRIN with angiogenesis, its value in prognosis, and its roles in RCC. Experimental Design EMMPRIN expression was examined in 50 RCC patients treated with radical nephrectomy. Angiogenesis, proliferation, and invasion activity were evaluated using EMMPRIN knockdown RCC cell lines. The size of EMMPRIN-overexpressing xenografts was measured and the degree of angiogenesis was quantified. EMMPRIN expression was evaluated in RCC patients who received sunitinib therapy and in sunitinib-resistant cells. Further, the relation between EMMPRIN expression and sensitivity to sunitinib was examined. Results EMMPRIN score was significantly associated with clinicopathological parameters in RCC patients, as well as being significantly correlated with microvessel area (MVA) in immature vessels and with prognosis. Down-regulation of EMMPRIN by siRNA led to decreased VEGF and bFGF expression, cell proliferation, and invasive potential. EMMPRIN over-expressing xenografts showed accelerated growth and MVA of immature vessels. EMMPRIN expression was significantly increased in patients who received sunitinib therapy as well as in sunitinib-resistant 786-O cells (786-suni). EMMPRIN-overexpressing RCC cells were resistant to sunitinib. Conclusion Our findings indicate that high expression of EMMPRIN in RCC plays important roles in tumor progression and sunitinib resistance. Therefore, EMMPRIN could be a novel target for the treatment of RCC.

Serum fucosylated haptoglobin as a novel prognostic biomarker predicting high-Gleason prostate cancer

Fucosylation is an oligosaccharide modification associated with cancer and inflammation, which is catalyzed by fucosyltransferases. Fucosylated haptoglobin (Fuc‐Hpt) has been identified as a novel biomarker for pancreatic cancer.

Microvessel area of immature vessels is a prognostic factor in renal cell carcinoma

To analyze the presence of immature vessels as a predictive factor of prognosis in patients with renal cell carcinoma.

Four cases of advanced renal cell carcinoma with pancreatic metastasis successfully treated with radiation therapy

We report four patients with pancreatic metastasis of renal cell carcinoma who were successfully treated with radiation therapy. The patients were one woman and three men with a median age of 55 years (range, 49 to 62 years) who underwent radical nephrectomy for primary renal cell carcinoma. The median interval from nephrectomy to the diagnosis of pancreatic metastasis was 129 months (range, 54 to 176 months). Two patients experienced melanorrhea and the other two were asymptomatic and diagnosed during standard follow up. In each patient, a total of 50 Gy in 2-Gy fractions over 5 weeks was prescribed, without any adverse events. All patients remain alive with a median follow up of 31 months (range, 11 to 81 months).

腎癌術後多発性骨転移に対しIL-2療法を施行中, 潰瘍性大腸炎様所見を伴う消化器症状を呈した1例

A 60-year-old man underwent retroperitoneal laparoscopic nephrectomy for left renal tumor (cT1bN0M0).The histopathological examination revealed Kidney cancer grade 3 pT1b. The following evaluations revealed multiple bone metastasis. The IFN-gamma with radiation therapy were performed. However the disease was progressive. So IL-2 70 million units per day 5 times a week started. The patient started to complain lower abdominal pain and watery diarrhea from administration day 28th. Blood test showed eosinophilia. At this point side effect of IL-2 therapy was suspected, then IL-2 was discontinued. But abdominal symptoms had continued. Consulting with a digestive physician, he diagnose as drug-induced colitis like ulcerative colitis by colon endoscopy. The symptoms were gradually improved by an antiallergic agent on our assumption that eosinophilia was concerned in this colitis. Many articles have reported that IL-2 was associated with the clinical mechanism of ulcerative colitis, but there seems no reports about such complications before. This case could suggest IL-2 relates to ulcerative colitis.

Bone marrow-derived cells contribute to regeneration of injured prostate epithelium and stroma.

Recent studies have reported that bone marrow‐derived cells (BMDCs), which are recruited to sites of tissue injury and inflammation, can differentiate into epithelial cells, such as liver, lung, gastrointestinal tract, and skin cells. We investigated the role of BMDCs in contributing to regeneration of injured prostate epithelium.

Abstract 2872: Serum fucosylated haptoglobin is a novel prognostic biomarker for prostate cancer: Detailed analyses of its oligosaccharide structures

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA Fucosylation is a crucial oligosaccharide modification in cancer and inflammation, which is catalyzed by alpha 1-3 (Futs3-7and Futs9-11)/ alpha 1-6(Fut8) fucosyltransferases. Alpha1-3 fucosyltransferase is involved in synthesis of Lewis type fucosylation and alpha 1-6 fucosylation is involved in core fucosylation. We have reported that fucosylated haptoglobin (Fuc-Hpt) is a cancer biomarker for patients with pancreatic cancer and colorectal cancer, and developed a lectin-antibody ELISA to measure serum levels of Fuc-Hpt. In the present study, we evaluated serum Fuc-Hpt levels in patients with localized prostate cancer and analyzed the molecular mechanisms in terms of the sources of Fuc-Hpt producing tissue. Methods: Fuc-Hpt levels in 98 preoperative serum samples from patients who underwent radical prostatectomy (RP) were measured by lectin-antibody ELISA. Expression of Fut8 and haptoglobin proteins in prostate cancer cell lines (LnCAP, 22Rv1, DU145, and PC3) and prostatectomy specimens were analyzed by Western blot and immunohistochemical analysis, respectively. Site-specific N-glycan analyses of haptoglobin in sera obtained from patients with prostate cancer were performed using liquid chromatography-electrospray ionization mass spectrometry. Results: Fuc-Hpt levels were significantly associated with Gleason score (GS), but not with PSA levels. The area under the receiver-operator characteristics curve (AUC) for Fuc-Hpt of the prediction of Gleason score 7 or more in prostatectomy specimens was 0.753, in contrast to the PSA AUC of 0.561 and the PSAD AUC of 0.558. The Fuc-Hpt AUC of the prediction of GS upgrading from biopsy GS6 to RP GS7 or more was 0.689, in contrast to the PSA AUC of 0.588 and the PSAD AUC of 0.557. Multivariable analysis revealed that Fuc-Hpt levels were significantly associated with biochemical recurrence after prostatectomy. Western blot analysis showed that prostate cancer cells expressed both Fut8 and haptoglobin. Immunohistochemical analysis showed positive Fut8 and haptoglobin staining in prostate cancer cells in prostatectomy specimens. Mass spectrometry analyses demonstrated that the oligosaccharide structure of Fuc-Hpt from men with prostate cancer was different from that from patients with gastrointestinal cancer. Conclusions: Fuc-Hpt in sera of patients with prostate cancer is mainly core Fuc-Hpt, but not Lewis type Fuc-Hpt. It is suggested that Fuc-Hpt is produced from prostate cancer cells. Elevations in serum Fuc-Hpt from patients with high Gleason score could be value in further studying Fuc-Hpt as a serum biomarker. Serum Fuc-Hpt levels may aid in prognostication of localized prostate cancer. Citation Format: Kazutoshi Fujita, Motohide Uemura, Wataru Nakata, Mototaka Sato, Akira Nagahara, Yasutomo Nakai, Mayuka Shimomura, Miyako Nakano, Eiji Miyoshi, Norio Nonomura. Serum fucosylated haptoglobin is a novel prognostic biomarker for prostate cancer: Detailed analyses of its oligosaccharide structures. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2872. doi:10.1158/1538-7445.AM2014-2872

Abstract 4359: TRAIL and Noxa are selectively up-regulated in prostate cancer cells downstream of the RIG-I/MAVS signaling pathway by non-replicating Sendai virus particles.

PURPOSE: The treatment of cancer with oncolytic viruses primarily depends on the selective viral replication in cancer cells. However, we previously reported that a replication-incompetent hemagglutinating virus of Japan (HVJ; Sendai virus) envelope (HVJ-E) suppressed the growth of human cancer cells, including prostate cancer cells, as effectively as replication-competent live HVJ without producing toxic effects in nonmalignant cells. Here, we analyze the molecular mechanism of the oncolytic activity of HVJ-E and also analyze the treatment efficacy of HVJ-E using an orthotopic prostate cancer model. EXPERIMENTAL DESIGN: The molecules responsible for HVJ-E-induced cancer cell death were elucidated in human prostate cancer cell lines, PC3, DU145 and LNCap. The cell viability was examined using MTS assay after HVJ-E treatment or the transfer of viral RNA genome fragments from HVJ-E. The viral RNA genome fragments and siRNAs were transferred to cells using liposome transfection. The expression of TNF-related apoptosis-inducing ligand (TRAIL) and Noxa were examined using Western blot analysis. The orthotopic prostate cancers were established by the inoculation of PC3 cells into the prostate of nonobese diabetic-severe combined immunodeficient (NOD-SCID) mice. After the tumors increased and microscopically visualized, HVJ-E was injected into the prostate of NOD-SCID mice. Tumor sections were prepared, and then, histological analysis and TUNEL assay were performed. RESULTS: The liposome-mediated transfer of viral RNA genome fragments from HVJ-E suppressed the viability of prostate cancer cells, PC3 and LNCaP, but not the viability of the noncancerous prostate epithelium, PNT2. Knockdown experiments using siRNAs showed that the cancer cell-selective killing induced by HVJ-E was mediated by retinoic acid-inducible gene I (RIG-I) and mitochondrial antiviral signaling protein (MAVS). Downstream of the RIG-I/MAVS pathway, both TRAIL and Noxa were up-regulated by either HVJ-E treatment or transfer of viral RNA genome fragments in PC3 cells but not in PNT2 cells. TRAIL siRNA and/or Noxa siRNA significantly inhibited HVJ-E-induced cell death in prostate cancer cells. Furthermore, the orthotopic prostate cancers were dramatically eradicated in NOD-SCID mice injected with HVJ-E. Although tubuloalveolar glands were histologically maintained in the prostate injected with HVJ-E, dramatic increase of TUNEL-positive apoptotic cells were detected exclusively in the tumors treated with HVJ-E. CONCLUSION: The oncolytic activity of HVJ-E was mediated by TRAIL and Noxa, downstream of the RIG-I/MAVS signaling pathway, selectively up-regulated in prostate cancer cells by HVJ-E treatment. The RIG-I/MAVS signaling pathway represents an attractive target for cancer therapy. Citation Format: Koji Hatano, Yasutomo Nakai, Taeko Matsushima-Miyagi, Motonari Nomura, Wataru Nakata, Takahiro Yoshida, Mototaka Sato, Atsunari Kawashima, Akira Nagahara, Kazutoshi Fujita, Motohide Uemura, Hitoshi Takayama, Yasufumi Kaneda, Norio Nonomura. TRAIL and Noxa are selectively up-regulated in prostate cancer cells downstream of the RIG-I/MAVS signaling pathway by non-replicating Sendai virus particles. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4359. doi:10.1158/1538-7445.AM2013-4359

1998 IMPACT OF HYPONATREMIA ON SURVIVAL OF PATIENTS WITH METASTATIC RENAL CELL CARCINOMA TREATED WITH MOLECULAR TARGETED THERAPY

Atsunari Kawashima*, Hitoshi Takayama, Suita, Japan; Yasuyuki Arai, Osaka, Japan; Mikio Nin, Sakai, Japan; Go Tanigawa, Yutaka Yasunaga, Osaka, Japan; Masatoshi Mukai, Toyonaka, Japan; Hironori Nomura, Daizo Oka, Toshiaki Yoshioka, Osaka, Japan; Satoko Fukuda, Ikeda, Japan; Kenji Nishimura, Nishinomiya, Japan; Nobukazu Murosaki, Itami, Japan; Minoru Koga, Minoh, Japan; Yasuyuki Kojima, Suita, Japan; Miyaji Kyakuno, Osaka, Japan; Takahiro Yoshida, Koji Hatano, Mototaka Sato, Motohide Uemura, Yasutomo Nakai, Suita, Japan; Kazuo Nishimura, Osaka, Japan; Akira Tsujimura, Norio Nonomura, Suita, Japan

Abstract 2475: Excision repair cross complementing group 1 (ERCC1) predicts the efficacy of chemoradiotherapy for invasive bladder cancer

Purpose: Chemoradiation therapy (CRT) is now widely recognized as bladder-preserving therapy for muscle-invasive bladder cancer (MIBC). However, some patients who fail CRT may miss the chance to be cured by cystectomy. Therefore, it is important to select patients with MIBC who are expected to have a good response to CRT. Several reports indicate that the excision repair cross-complementing group 1 (ERCC1) gene is associated with resistance to cisplatin and radiation therapy. In this study, we examined the correlation between ERCC1 and CRT in vitro and in vivo in bladder cancer. Experimental Design: Bladder cancer cell lines T24, 5637, Cl8-2 (multi-drug-resistant subline of T24), and CDDP10-3 (cisplatin-resistant subline of T24) were used for in vitro assays to measure ERCC1 expression level and growth inhibition with cisplatin or irradiation (IR). To clarify the association between ERCC1 and cisplatin resistance in bladder cancer cells, we knocked down ERCC1 in Cl8-2 and CDDDP10-3 with siRNA (C18-2ΔERCC1 and CDDP10-3ΔERCC1). To further prove the cause of the radiation sensitivity of ERCC1 knockdown cells, we measured the phosphorylated histone variant H2A.X, a marker of DNA damage. In the clinical study, we then examined by immunohistochemistry whether ERCC1 nuclear staining correlates with the efficacy of CRT using cisplatin in 22 patients with MIBC. Results: Cl8-2 cells expressed ERCC1 mRNA 5.96-fold higher than did T24. Cl8-2 and CDDP10-3 were more resistant to cisplatin or IR than was T24. Our ERCC1 knockdown experiments showed that there was statistical difference in the resistance to IR exposure not cisplatin compared with C18-2CTL and CDDP10-3CTL. C18-2ΔERCC1 and CDDP10-3ΔERCC1 recovered more slowly in terms of the number of phospho-H2A.X foci than did C18-2cont and CDDP10-3cont, suggesting continued accumulation or persistence of DSBs. In immunohistochemistry with ERCC1, six of eight positive cases did not have complete response to CRT, whereas 12 of 14 negative cases had complete response. Sensitivity and specificity were 75% and 85.7%, respectively (p = 0.008). Conclusion: Our results suggest that in some bladder cancer cells, ERCC1 expression correlates with IR resistance but not with cisplatin resistance. Moreover, the lack of ERCC1 expression correlated well with the efficacy of CRT, and especially with that of IR, in our clinical study. Although further study is needed, ERCC1 expression level may predict the efficacy of CRT for MIBC. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2475. doi:10.1158/1538-7445.AM2011-2475