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Bile acids have been shown to be important regulatory molecules for cells in the liver and gastrointestinal tract. They can activate various cell signaling pathways including extracellular regulated kinase (ERK)1/2 and protein kinase B (AKT) as well as the G-protein-coupled receptor (GPCR) membrane-type bile acid receptor (TGR5/M-BAR). Activation of the ERK1/2 and AKT signaling pathways by conjugated bile acids has been reported to be sensitive to pertussis toxin (PTX) and dominant-negative Gα(i) in primary rodent hepatocytes. However, the GPCRs responsible for activation of these pathways have not been identified. Screening GPCRs in the lipid-activated phylogenetic family (expressed in HEK293 cells) identified sphingosine-1-phosphate receptor 2 (S1P(2) ) as being activated by taurocholate (TCA). TCA, taurodeoxycholic acid (TDCA), tauroursodeoxycholic acid (TUDCA), glycocholic acid (GCA), glycodeoxycholic acid (GDCA), and S1P-induced activation of ERK1/2 and AKT were significantly inhibited by JTE-013, a S1P(2) antagonist, in primary rat hepatocytes. JTE-013 significantly inhibited hepatic ERK1/2 and AKT activation as well as short heterodimeric partner (SHP) mRNA induction by TCA in the chronic bile fistula rat. Knockdown of the expression of S1P(2) by a recombinant lentivirus encoding S1P(2) shRNA markedly inhibited the activation of ERK1/2 and AKT by TCA and S1P in rat primary hepatocytes. Primary hepatocytes prepared from S1P(2) knock out (S1P(2) (-/-) ) mice were significantly blunted in the activation of the ERK1/2 and AKT pathways by TCA. Structural modeling of the S1P receptors indicated that only S1P(2) can accommodate TCA binding. In summary, all these data support the hypothesis that conjugated bile acids activate the ERK1/2 and AKT signaling pathways primarily through S1P(2) in primary rodent hepatocytes.

Sphingosine-1-phosphate (S1P) is a pleiotropic bioactive lipid mediator that regulates several processes important for hematologic cancer progression. S1P is generated by two sphingosine kinases, SphK1 and SphK2, and is exported outside the cell, where it activates specific cell surface S1P G-protein coupled receptors in autocrine/paracrine manner, coined “inside-out signaling”. In this review, we highlight the importance of SphK1 and inside-out signaling by S1P in hematologic malignancy. We also summarize the results of studies targeting the SphK1/S1P/S1P receptor axis and the effects of the S1P receptor modulator, FTY720, in hematologic malignancy.

In MKN1 gastric cancer cells, lysophosphatidic acid (LPA) upregulates expression of sphingosine kinase 1 (SphK1) and its downregulation or inhibition suppresses LPA mediated proliferation. Although LPA activates numerous signaling pathways downstream of its receptors, including extracellular-signal-regulated kinase 1/2, p38, JNK, and Akt, and the transactivation of the epidermal growth factor receptor, pharmacological and molecular approaches demonstrated that only activation of ERK1, in addition to the CCAAT/enhancer-binding protein β transcription factor, is involved in transcriptional upregulation of SphK1 by LPA. Our data implicate ERK1 as an important mediator of LPA signaling leading to upregulation of SphK1 and point to SphK1 and sphingosine-1-phosphate production as potential therapeutic targets in gastric cancer.

e13111Background: Breast cancer is the most frequent cancer among women both in US and Japan. Triple negative breast cancer (TNBC), a subset of breast cancers that are negative for estrogen recepto...

e16068Background: Defective homologous recombination (HR) due to genetic aberrations of HR genes contributes to a high response of platinum therapy in ovarian and breast cancers. The aim of this st...

e15647Background: Hypermutation, which is a molecular subtype with high tumor mutation burden (TMB), has been recognized as a biomarker of anti-PD-1 therapy in many cancers. Hypermutation can be de...

Background: Sphingosine-1-phosphate (S1P), a bioactive sphingolipid mediator that is generated by sphingosine kinase 1 (SphK1) when it is phosphorylated (pSphK1) inside cells, has been implicated in regulation of many process important for breast cancer progression. Previously we have shown that S1P is exported out of human breast cancer cells by ATP-binding cassette (ABC) transporter ABCC1, but not by ABCB1, both known multidrug resistance proteins that efflux chemotherapeutic agents. However, the pathological consequences of these events to breast cancer progression and metastasis have not been elucidated. Here, we report that high expression of ABCC1, but not ABCB1, is associated with poor prognosis in breast cancer patients via exporting S1P. Materials and methods: Microarray based gene expression data of 2509 patients associated with their survival were obtained from METABRIC database. Single gene survival analysis based on expressin of SphK1, and dual ABCC1 or ABCB1 and SphK1 survival analyses were perfomerd. For protein analyses, tissues were obrained from 275 patients with stage 1-3 breast cancers treated in Yokohama City University Medical Center in Japan between 2006 and 2008. The expression of pSphK1 was analyzed by immunohistochemistry and investigate the relationship with clinicopathological findings. For in vitro and in vivo experiments, breast cancer cell lines were transfected by ABCB1, ABCC1 or vector transiently or stably. BALB/c nu/nu mice and BALB/c mice were used for in vivo experiments. S1P was measured by LC-ESI-MS/MS. Results: SphK1 expression significantly associate with worse overall survival (median survival of 124 months with high SphK1 expression compared to 163 months for patients with low SphK1 expression, p=0.0014). Although patients with high ABCC1 expression had only a slightly worse overall survival of 150 months, those with high levels of both SphK1 and ABCC1 had much worse prognosis with median overall survival of 114 months (p < 0.0068). Such association was not observed with ABCB1 expression. The frequency of strong pSphK1 protein expression was higher in HER2 enrhiched or TNBC than in Luminal. pSphK1 was more prevalent and increased in a larger tumors and in tumors from patients with lymph node metastases. Patients with breast cancers that express both pSphK1 and ABCC1 proteins have significantly shorter disease free survival. Overexpression of ABCC1, but not ABCB1, in human MCF7 and murine 4T1 cells enhanced S1P secretion, proliferation and migration of breast cancer cells. Implantation of breast cancer cells overexpressing ABCC1, but not ABCB1, into the mammary pad markedly enhanced tumor growth, angiogenesis and lymphangiogenesis with concomitant increases in lymph node and lung metastases as well as shorter survival of mice. Interestingly, S1P exported via ABCC1 from breast cancer cells upregulated transcription of SphK1 and its own formation. Conclusions: Our findings suggest that production and export of S1P via ABCC1, but not ABCB1, is associated with worse overall and disease free survival of breast cancer patients and that S1P axis play a role in aggressive biology of breast cancer progression and metastasis. Citation Format: Yamada A, Nagahashi M, Aoyagi T, Huang W-C, Lima S, Miyazaki H, Narui K, Ishikawa T, Endo I, Waters MR, Milstien S, Spiegel S, Takabe K. Sphingosine-1-phosphate produced by sphingosine kinase 1 and exported via ABCC1 shortens survival of mice and humans with breast cancer [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P5-03-05.

Background: The bioactive lipid mediator sphingosine-1-phosphate (S1P) has emerged as a key regulatory molecule in cancer progression. We previously demonstrated that S1P is a crucial mediator of breast cancer-induced angiogenesis and lymphangiogenesis, and promote metastasis. Although increasing number of in vitro and in vivo experiments have revealed the importance of S1P in cancer progression, the data on the roles of S1P in human patients are very limited. The aim of this study is to reveal the clinical relevance of S1P in the interaction between cancer and the tumor microenvironment by examining the levels of the sphingolipids in patient breast cancer tissue samples. Material and Method: Breast cancer tissue, peri-tumor tissue, and normal breast tissue were collected from 20 breast cancer patients immediately after surgery that were conducted from November 2015 to February 2016 at Niigata University Medical and Dental Hospital. Sphingolipids were quantified by liquid chromatography–electrospray ionization tandem mass spectrometry. The expression level of each enzyme-encoding gene involved in S1P production was evaluated by retrieving RNA sequencing and gene expression quantification data from breast cancer tissues (n = 112) and paired normal breast tissues (n = 112) using the Genomics Data Commons (GDC) data portal of the The Cancer Genome Atlas (TCGA) cohort. Gene expression levels were derived using normalization methods provided in the DESeq2 package. Result: The levels of the sphingolipids sphingosine (Sph), dihydro-sphingosine (DHSph), S1P, and dihydro-S1P (DHS1P) were successfully determined in breast cancer, peri-tumor, and normal breast tissues from all of the 20 patients. As expected, a one-way ANOVA revealed that S1P levels were significantly different depending on the location (F(2,57) = 7.029, P = 0.002). The Tukey post hoc test revealed that S1P levels in tumors were significantly higher than those in normal breast tissue and peri-tumor tissue (P < 0.05). Similarly, Sph and DHSph levels in tumors were significantly higher than in normal breast tissue and peri-tumor tissue. Both SPHK1 and SPHK2 gene expression levels in breast cancer tissue were higher than those in normal breast tissue. Interestingly, expression of some of the S1P-related genes; S1PR3, ABCC1, SGPL1, and ORMDL2, were significantly increased in the breast cancer tissue compared to normal breast tissue. On the other hand, there was significantly decreased expression of the S1P-related genes S1PR1, S1PR2, ABCG2, SPNS2, SGPP1 and ORMDL3, in breast cancer tissue compared to normal breast tissue. Conclusion: We demonstrated that the major source of S1P is the tumor tissue, and not the peri-tumor tissue despite the fact that angiogenesis and lymphangiogenesis are occurring more in the peri-tumor area, which implicate that S1P may have further role inside the tumor. Our results indicated the complexity of S1P signaling in human cancer than expected based on the results in vivo experiments. Citation Format: Tsuchida J, Nagahashi M, Nakajima M, Takabe K, Wakai T. The levels of sphingosine-1-phosphate and its related gene expressions in breast cancer patients [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P1-01-21.

Introduction: Obesity with associated inflammation is now recognized as a risk factor for breast cancer and increased incidence of distant metastases. However, the link between obesity and breast cancer progression remains poorly understood. There is growing evidence that sphingosine-1-phosphate (S1P), a pleiotropic bioactive sphingolipid metabolite enriched both in blood and lymphatic fluid is involved in inflammation, obesity, and breast cancer progression. Our hypothesis is that obesity increases levels of S1P in both tumor and its microenvironment, which play a role in obesity-induced inflammation and breast cancer metastasis. The aim of this study is to test this hypothesis in in vitro and in vivo as well as patient settings. Methods: Levels of sphingolipids including S1P in serum from breast cancer patients were quantified. Orthotopically-implanted E0771 syngeneic breast cancer and MMTV-PyMT transgenic breast cancer mouse models were used. Mice were fed with normal or high-fat diet (HFD). FTY720 was administered orally (1 mg/kg/day). To examine pre-metastatic niche formation, a mouse model utilizing tail vein injection of E0771 cells was used. In this model, mice were treated with conditioned media from E0771 breast cancer cells overexpressing SphK1 (K1-CM) or that from E0771 cells cultured with the vector control (CT-CM), prior to tail vein injections of naive E0771 cells. S1P levels were determined by electrospray ionization-tandem mass spectrometry. Results: We found that obesity significantly increased S1P levels in serum from breast cancer patients. In animal breast cancer models, HFD upregulated expression of sphingosine kinase 1 (SphK1), the enzyme that produces S1P, and its receptor S1PR1 in syngeneic and spontaneous breast tumors. HFD also significantly increased S1P in breast tumors and in the tumor interstitial fluid, which is a component of the tumor microenvironment and bathes cancer cells in the tumor. Targeting the SphK1/S1P/S1PR1 axis with FTY720/fingolimod attenuated obesity-induced key pro-inflammatory cytokines, macrophage infiltration, and tumor progression. In addition, S1P produced by tumor SphK1 primed lung pre-metastatic niches, increased macrophage recruitment into the lung, and induced IL-6 and signaling pathways important for lung metastatic colonization. FTY720 suppressed HFD-induced lung IL-6 and macrophage infiltration as well as S1P-mediated signaling pathways and dramatically reduced formation of metastatic foci. In tumor bearing mice, FTY720 also suppressed obesity-related inflammation, S1P signaling, pulmonary metastasis, and prolonged survival. Conclusion: Our results highlight a critical role for circulating S1P produced by tumor and the SphK1/S1P/S1PR1 axis in obesity-related inflammation, metastatic niche formation and breast cancer metastasis and suggest that targeting the SphK1/S1P/S1PR1 axis would be a useful therapeutic for obesity promoted metastatic breast cancer. Citation Format: Nagahashi M, Yamada A, Aoyagi T, Huang W-C, Terracina KP, Hait N, Allegood JC, Tsuchida J, Nakajima M, Katsuta E, Milstien S, Wakai T, Spiegel S, Takabe K. Targeting the SphK1/S1P/S1PR1 axis that connects obesity, chronic inflammation, and breast cancer metastasis [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P1-01-06.

BACKGROUND: We tested the hypothesis that in patients with T1 extrahepatic cholangiocarcinoma (EHC), prognosis postresection is significantly different for those with tumors that are limited to the mucosa than for those with tumors that have invaded (but not penetrated) the fibromuscular layer. METHODS: A retrospective analysis was conducted of 33 consecutive patients with pathologic T1 (pT1) EHC tumors. According to the depth of invasion, the pT1 tumors were divided into 2 groups: Group 1, tumors that were limited to the mucosa (mucosal tumors); and Group 2, tumors that had invaded (but not penetrated) the fibromuscular layer (fibromuscular layer-invasive tumors). Long-term outcomes after resection were compared between the 2 groups for a median follow-up time of 175 months. RESULTS: Eighteen patients had mucosal tumors and 15 patients had tumors that had invaded the fibromuscular layer. None of the patients with mucosal tumors had lymphovascular invasion, whereas 3 of the patients with fibromuscular layer-invasive tumors had lymphovascular invasion (P = .083). Overall survival after resection was better in Group 1 than in Group 2 (cumulative 10-year survival rate, 100% vs 52%; P = .024). The rate of disease-free survival after resection was higher in Group 1 than in Group 2 (cumulative disease-free 10-year survival rate, 100% vs 56%; P = .022). CONCLUSIONS: The long-term outcome after resection for EHC is significantly better for patients with mucosal tumors than for patients with fibromuscular layer-invasive tumors. This suggests that the depth of tumor invasion affects the postresection prognosis for patients with pT1 EHC. Cancer 2010. © 2010 American Cancer Society.

This study aimed to clarify the mode of hepatic spread from gallbladder carcinoma and to elucidate its prognostic value. A retrospective analysis was conducted of 42 consecutive patients who underwent resection for gallbladder carcinoma with hepatic involvement verified histologically. The mode of hepatic spread was classified into 3 patterns: direct invasion through the gallbladder bed, portal tract invasion, and hepatic metastatic nodules. Intrahepatic lymphatic invasion was declared when either single tumor cells or cell clusters were clearly visible within vessels that showed immunoreactivity to the D2-40 monoclonal antibody. Seven, 24, and 11 patients had direct invasion alone, portal tract invasion with (22 patients) or without (2 patients) direct invasion, and hepatic metastatic nodules, respectively. Of the 24 patients with portal tract invasion, 14 had intrahepatic lymphatic invasion, 8 had neither intrahepatic lymphatic nor venous invasion, and 2 had both intrahepatic lymphatic and venous invasion. To date, 4 patients with direct invasion alone and 4 patients with portal tract invasion survived more than 5 years after resection, whereas all the patients with hepatic metastatic nodules died within 11 months after resection, irrespective of the type of hepatectomy. The mode of hepatic spread (P

We report a case of a 29-year-old male patient with a generalized adenovirus (AdV) infection after cord blood transplantation (CBT) for acute myelocytic leukemia with maturation at 2nd complete remission. Before engraftment, hemorrhagic cystitis was caused by AdV, which resulted in hydronephrosis, renal failure, and adenoviremia on day 34. Forced diuresis, hemodialysis, withdrawal of cyclosporin A, and administration of gamma-globulin or vidarabine were not effective and the patient died of pulmonary alveolar hemorrhage on day 67. At autopsy, old inflammatory change only was observed in the bladder section. In the lungs and kidneys, granular deposits in the nucleus and a high copy number of AdV-DNA were observed. Molecular diagnosis using PCR-restriction fragment length polymorphism analysis demonstrated that AdV with the serotype 14 caused the cystitis. However, retrospective genome typing using PCR sequencing revealed the infection of AdV serotype 35 in the kidneys, lungs, and serum. The present case suggested that Adv infection could not be always caused by a single AdV serotype, and suggested that multiple serotype infection was very difficult to treat. It is desired that a consensus regarding the treatment of AdV infections is established.

症例は60歳の男性で, 家族歴として, 一卵性双生児の兄が家族性大腸腺腫症で加療中である. 1974年, 家族性大腸腺腫症と診断され, 結腸全摘術, 回腸直腸吻合術を施行された. 1991年, 残存直腸の多発ポリープに対し残存直腸切除, 回腸. 肛門吻合術を施行された. 2000年, 多発胃癌に対し, 幽門側胃切除術を施行, 2002年, 残胃癌に対し内視鏡的粘膜切除術, 2006年, 局所切除術を施行された. 2007年3月, 上部消化管内視鏡検査で食道胃接合部直下に0-IIa病変を認め, また十二指腸に密生する多発ポリープを認めた. いずれも生検で高分化腺癌と診断された. 5月下旬, 胃局所切除術, 膵頭温存十二指腸全摘術を施行した. 病理組織学的診断は, 胃は高分化腺癌, 十二指腸は高分化腺癌および腺腫の混在する病変で, いずれも粘膜内癌であった. 術後, 縫合不全, 膵液瘻, 胆汁瘻などの合併症なく順調に経過し, 第11病日に退院した.

AIM: To disclose geographic differences in genetic changes involved in gallbladder carcinogenesis between two distinct high-incidence areas of Japan and Hungary. METHODS: We examined 42 cases of gallbladder carcinoma: 22 Japanese and 20 Hungarian cases. p53 mutations at exons 5 to 8 and K-ras mutations at codon 12 were tested by direct sequencing. Microsatellite instability was determined from fluorescent dye-labeled PCR amplifications of five-microsatellite markers (BAT-25, BAT-26, D2S123, D5S346, and D17S250). RESULTS: Mutations of p53 were detected in 11 of 22 Japanese cases and 6 of 18 Hungarian cases (11/22 vs 6/18, P = 0.348). Transition at CpG sites was found in none of 11 Japanese cases and 2 of 6 Hungarian cases; the difference was marginally significant (0/11 vs 2/6, P = 0.110). K-ras mutations were detected in only one of the Hungarian cases. Eight of 19 (42.1%) Japanese cases were MSI-high (presence of novel peaks in more than one of the five loci analyzed), whereas only 1 of 15 (6.7%) Hungarian cases was MSI-high (P = 0.047). CONCLUSION: It appears that the p53 mutations and MSI differ in patients with gallbladder carcinoma between two distinct high-incidence areas. Geographic variation might exist in the process of gallbladder carcinogenesis.

大動脈周囲リンパ節転移陽性胆嚢癌症例の予後は極めて不良である. 今回, 大動脈周囲リンパ節転移陽性で切除後5年以上生存した胆嚢腺扁平上皮癌の1例を経験したので報告する. 症例は62歳の男性で, 進行胆嚢癌に対して胆嚢床切除+膵頭十二指腸切除+大動脈周囲リンパ節郭清を施行した. 組織学的には腺扁平上皮癌で, 所属リンパ節4個 (うち大動脈周囲リンパ節1個) に転移を認めた. 術後29か月目のCTで大動脈周囲リンパ節再発を認め, 同領域の郭清を施行した. その後, 軟部組織 (右膝) 転移, 骨転移 (第2腰椎) が出現し, 右下肢切断, 放射線照射を施行した. 初回手術から60か月後には左鎖骨上窩リンパ節転移が出現し郭清を施行したが, 多発肺転移が出現し, 初回手術から77か月後に原病死した. 本症例の経験および文献的考察から, 胆嚢癌では大動脈周囲リンパ節転移に対する郭清が有効な症例もまれながら存在することが示された.

Background Long-term graft survival of partial pancreas auto-transplantation after total pancreatectomy has not been clarified. The clinical implications of repeat completion pancreatectomy for locally recurrent pancreatic carcinoma in the remnant pancreas after initial pancreatectomy also have not been clarified. Methods We have previously reported a 61-year-old woman presenting with re-sectable carcinoma of the remnant pancreas at 3 years after undergoing a pylorus-preserving pancreaticoduodenectomy for invasive ductal carcinoma of the pancreas head. We also performed distal pancreas auto-transplantation with the use of a part of the resected pancreas to preserve endocrine function. Results The patient was discharged at 20 days after surgery without any complications. She had been followed regularly in our outpatient clinic. She had been treated with S-1 as adjuvant chemotherapy; 72 months after the completion total pancreatectomy with distal partial pancreas auto-transplantation, the patient was alive without any evidence of the pancreatic carcinoma recurrence. The pancreas graft was still functioning with a blood glucose level of 112 mg/dL, HbA1C of 6.7%, and serum C-peptide of 1.2 ng/mL; and urinary C-peptide was 11.6 μg/d. Conclusions Our patient demonstrated that repeated pancreatectomies can provide a chance for survival after a locally recurrent pancreatic carcinoma if the disease is limited to the remnant pancreas. An additional partial pancreas auto-transplantation was successfully performed to preserve endocrine function. However, the indications for pancreas auto-transplantation should be decided carefully in the context of pancreatic carcinoma recurrence.

Background Complete immune tolerance is the chief goal in organ transplantation. This study aimed to evaluate patients who successfully withdrew from immunosuppressive (IS) agents after living donor liver transplantation (LDLT). Materials and Methods A retrospective review of all adult LDLT from July 1999 to March 2012 was conducted. In patients who acquired immune tolerance after LDLT, their background and the course of surgical procedures were evaluated. Results Of a total of 101 adult LDLT patients, 8 patients were completely free of IS agents. Six of these patients (75%) were female, and the median age at the time of transplantation was 56 years (range, 31–66 years). The primary disease causing liver failure was type C liver cirrhosis (50%), fulminant hepatitis (25%), type B liver cirrhosis (12%), and alcoholic liver cirrhosis (12%). The median Child-Pugh score and MELD score were 13 points (range, 8–15 points) and 19 points (range, 10–18 points), respectively. The living related donor was the recipient's child (75%), sibling (12%), or parent (12%). ABO compatibility was identical in 62%, compatible in 25%, and incompatible in 12%. Conclusions In this study, we evaluated the adult patients who successfully withdrew from IS agents after LDLT. In most cases, it took more than 5 years to reduce IS agents. Because monitoring of the serum transaminase level is not adequate to detect chronic liver fibrosis in immune tolerance cases, further study is required to find appropriate protocols for reducing IS agent use after LDLT.

Pneumatosis cystoides intestinalis (PCI) is a relatively rare condition, characterized by subserosal or submucosal air within the bowel wall. Herein, we report a rare case of PCI secondary to treatment with an epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI). A 71-year-old man, who had received gefitinib therapy for 2 years and 5 months for lung adenocarcinoma with metastases to the bones and brain, presented with abdominal pain, diarrhea, and vomiting. Computed tomography of the abdomen revealed intramural air in the small bowel, free air in the abdomen, and moderate ascites. A diagnosis of PCI was made, and the patient was managed conservatively by discontinuing gefitinib treatment, because his vital signs were stable and there was no sign of peritonitis. The patient's symptoms gradually improved, and follow-up CT after 1 week revealed that the initial findings had almost completely resolved. Clinicians should note that treatment with gefitinib might cause PCI.

It has now become clear that the complex interplay of cancer and the immune responses against it plays a critical role in the tumor microenvironment during cancer progression. As new targets for cancer treatment are being discovered and investigated, murine models used for preclinical studies need to include intact immune responses to provide a closer correlation with human cancer. We have recently developed a modified syngeneic orthotopic murine colon cancer model that mimics human colon cancer progression with consistent results.Tumors were created using the murine colon adenocarcinoma cell line, CT26, modified to overexpress the firefly luciferase gene (CT26-luc1), which allowed real-time in vivo monitoring of tumor burden when the substrate, D-luciferin, was injected intraperitoneally using the In Vivo Imaging System. Mice are Balb/c (Harlan), syngeneic with the CT26-luc1 cells. Cells are injected submucosally, suspended in Matrigel, into the cecum wall under direct visualization.The model has demonstrated consistent implantation in the cecum. In vivo bioluminescence allowed real-time monitoring of total tumor burden. Perioperative preparation had a significant impact on reproducibility of the model. Finally, total tumor burden quantified with bioluminescence enabled estimation of lymph node metastasis ex vivo.This method maintains an intact immune response and closely approximates the clinical tumor microenvironment. It is expected to provide an invaluable murine metastatic colon cancer model particularly in preclinical studies for drug development targeting those mechanisms.

We report the case of a 72-year-old female who was admitted to our hospital because of obstructive colitis. Blood analysis showed her serum carcinoembryonic antigen (CEA) level to be 156.0 ng/mL. A sigmoidectomy and descending colostomy were performed for obstructive colitis due to colonic diverticulitis. Histopathological examination revealed active inflammation of the sigmoid colon without neoplasia. Her serum CEA level decreased within normal limits immediately after surgery.

症例は48歳の男性. HBs抗原陽性でAFPが高値を示し,左肝管内に3cm大の腫瘍が存在,左肝内胆管の拡張を認めた.精査にて胆管内発育型の肝細胞癌ないし胆管癌が考えられた.手術を予定していたが,急性腹症を発症,経皮経肝胆嚢ドレナージ(PTGBD)を行うと血性胆汁で,胆管内腫瘍の出血と判断した.後日,肝左葉切除,胆管切除を施行した. S2に2 cm大の腫瘍を認め,これより胆管内に浸潤.乳頭状に発育した5 cmの腫瘍が左肝管を閉塞していた.病理は肝細胞癌であった.本症例は肝細胞癌が胆管浸潤,胆管内発育し,腫瘍壊死により胆道出血をきたしたと考えられる.肝細胞癌の胆管への浸潤・発育あるいは胆道出血はいずれも比較的稀であり,文献的考察を加え報告する.

胃癌の壁内転移は極めて稀である.今回われわれは残胃に発生した胃癌の壁内転移の1例を経験したので報告する.症例は63歳,男性.胃癌の診断で当科を紹介され, 2002年9月,幽門側胃切除術を施行した.病理診断は乳頭腺癌, ss, ly3, v3, n0であった.術後3カ月, CA19-9が上昇.腹部CTで多発性肝転移の診断でTS-1,タキソールによる化学療法を行った.しかし肝転移は増大.また,胃内視鏡検査で,残胃内に山田III型腫瘍を3個認め,残胃への壁内転移が疑われた.胃の腫瘍による消化器症状が強いこと,全身化学療法で肝転移が悪化したことより手術の方針とし, 2003年8月,残胃全摘術,肝動注カテーテル留置術を行った.残胃内に腫瘍を3個認め,いずれも原発巣と同じ乳頭腺癌であった.術後経過は良好であり, 19病日に退院した.

A 53-year-old Japanese man with fulminant myocarditis was referred. Percutaneous cardiopulmonary support (PCPS) was introduced immediately and intravenous immunoglobulin (IVIG) therapy followed for 2 days. Cardiac function showed signs of recovery on the 4th hospital day and the patient was weaned from PCPS on the 7th hospital day. Creatine kinase-MB peaked at 12 h after admission and was 176 ng/ml. Endomyocardial biopsy showed active myocarditis. A marked increase of the neutralizing antibody titer suggested coxsackievirus B3 infection. Plasma concentrations of cytokines and neurohumoral factors were analyzed. Proinflammatory cytokines, such as interleukin (IL)-1beta, IL-6 and tumor necrosis factor (TNF-alpha), and anti-inflammatory cytokines, such as IL-1 receptor antagonist, soluble TNF receptor-1 and IL-10, were elevated on admission and all had decreased on the 7th hospital day. Brain natriuretic peptide and noradrenaline were already elevated upon admission (1,940 pg/ml and 4.6 ng/ml, respectively) and decreased thereafter. Although IVIG therapy under PCPS is a common treatment for fulminant myocarditis, the immunological response in vivo remains unclear. This case demonstrated suppression of serum cytokines after IVIG and PCPS treatment. Immunological parameters in those who have been treated with IVIG and PCPS and survived without complications are of great value for evaluation of the therapy. Further analysis with more cases in a multicenter study is necessary.

症例は62歳の男性で,黄疸を主訴として来院.経皮経肝胆道造影検査にて上中部胆管の完全閉塞を認め,腹部CT検査で肝外胆管に直径2cmの腫瘤を認めた.胆汁細胞診で腺癌細胞が証明され,上中部胆管癌と診断.幽門輪温存膵頭十二指腸切除術, D2郭清を施行した.病理組織学的検査で, HE染色では大部分が低分化型腺癌からなり,免疫組織学的染色でchromogranin Aが約10分の1で陽性のため,胆管腺内分泌細胞癌と診断した.術後補助化学療法は施行せず.外来にて経過観察中, 5カ月目にCT検査を行ったところ,肝両葉に多発性肝転移を認めた.その後も肝転移が急速に進行し術後約7カ月で死亡した.稀な肝外胆管原発の腺内分泌細胞癌の切除例を経験したので報告する.

e15592 Background: Molecular heterogeneity represents a significant hurdle in realizing precision medicine for gastric cancer (GC). Large-scale whole exome sequencing projects have identified distinct molecular subtypes to help define the heterogeneity of GC. However, it remains unclear whether the targeted gene panel-based sequencing can provide optimum targeted therapies and clinical utility in GC. The aim of this study is to generate comprehensive genomic profiling data and classify Japanese GC into actionable clusters associated with targeted therapies. Methods: FFPE tumor tissues were obtained from surgical or biopsy specimens of 207 Japanese patients with GC. Extracted DNA was subjected to genomic sequencing for 435 cancer related genes including 69 druggable genes with FDA approved targeted therapies. Somatic mutations, copy number alterations (SCNA), microsatellite instability (MSI) and Epstein-Barr virus (EBV) infection were evaluated using sequencing data. Results: Genomic sequencing identified at least one alteration of 435 genes in 194 pts (94%), and that of 69 druggable genes in 141 pts (68%). The most frequently altered druggable gene was ERBB2 (14%), following BRCA2 (11%) and ATM (10%). We successfully classified 207 tumors into four molecular subtypes, similar to the previously report; EBV (4%), MSI (8%), chromosomal instability (58%) and genomically stable subtype (30%). Frequent alterations of druggable genes ( > 5%) were widely observed through these subtypes. To discover the novel classifications associated with targeted therapies, we classified 207 tumors using mutation rate and hierarchical clustering. We identified a hypermutated group (n = 32), and a remaining non-hypermutated group (n = 175) which were sub-divided into six clusters including five actionable ones; ERBB2 (n = 25), CDKN2A and CDKN2B (n = 10), KRAS (n = 10), BRCA2 (n = 9) and ATM cluster (n = 12). Interestingly, we experienced a case of unresectable GC with a remarkable response for anti-HER2 therapy in the ERBB2 cluster. Conclusions: Genomic sequencing using a 435-gene panel has the potential to provide the information of optimum targeted therapies for upcoming precision medicine in Japanese GC.

e23122 Background: Breast cancer is the leading cancer among women both in US and Japan. Triple negative breast cancer (TNBC) is the subset of breast cancers that are negative for estrogen receptor (ER), progesterone receptor, and HER2. While the selective ER modulator tamoxifen or anti-HER2 therapy for breast cancer patients with ER positive or HER2 protein overexpression are the most successful examples of targeted therapies, only limited therapies are available for patients with TNBCs, which are associated with a poor prognosis. Advance in next-generation sequencing enables us to identify actionable driver mutations that can be potentially treated by targeted therapies in each cancer patient. The aim of this study is to examine actionable driver mutations in TNBCs in Japan by comprehensive genomic sequencing (CGS) with 435 gene panel, and compare the driver events in Japan with TCGA database to validate the utility of CGS. Methods: We examined all exons of 435 known cancer genes in Japanese TNBC patients (N = 53) by CGS and evaluated for concordance among independent data obtained from The Cancer Genome Atlas-TNBC whole exome sequencing database (N = 123). Results: Oncogenic driver mutations were identified in 51 of 53 Japanese patients (96%) with TNBC and 36 of 53 patients (67%) harbored mutations in genes associated with FDA-approved targeted therapies, indicating the potential clinical utility of a large gene panel for evaluating patients with TNBC. Among 80 total genetic alterations, frequently mutated genes ( > 10% patients) were TP53, PIK3CA and PTEN. Overall, the mutation spectrum of the Japanese patients is similar to that of the TCGA population, except amplification of MYC. Conclusions: Use of a CGS panel of 435 genes can reliably identify all of the critical mutations in TNBC patients, which are similar as TCGA data. The information derived from CGS can be used to determine the optimal treatment for TNBC patients.

e23191 Background: Microsatellite instability-high status (MSI-H) and alterations in the DNA mismatch repair pathway associate with the efficacy of 5-FU and immune checkpoint inhibitors in patients with gastrointestinal cancers. The activin type II receptor (ACVR2) that binds to the transforming growth factor beta superfamily of ligands is frequently mutated in MSI-H colorectal cancer. However, the incidence of ACVR2 mutations in gastric cancer patients remains unclear. The aim of this study is to reveal the incidence and to examine the association between the MSI-H and ACVR2A mutations in gastric cancer patients. Methods: 124 archived FFPE gastric cancer tissues (stage I-IV), who were operated at Niigata University Medical and Dental Hospital or Niigata Cancer Center Hospital, were analyzed for ACVR2A mutation and MSI status with the NGS-based comprehensive genomic test platform. Clinicopathological characteristics of the patients were also examined. Results: All 124 gastric cancer patients were successfully analyzed. 13 out of 124 patients (10.4%) showed MSI-H status. Interestingly, 10 of 13 MSI-H patients (76.9%) showed ACVR2A mutation, where none (0%) was found among patients with microsatellite stable status (P < 0.001), indicating the strong association between ACVR2A mutation and MSI status in gastric cancer patients. In the ACVR2A mutated group, there was a female predominance (P < 0.05), and cancers of the lower part of the stomach were more common (P < 0.05), compared with the wild type group. Only one of 10 patients with ACVR2A mutation died, and the patients with ACVR2A mutation show a 5-year overall survival rate of 90%. No statistically significant difference in survival was achieved between patients with ACVR2A mutation and wild type; this is probably due to the small number of patients. Conclusions: 10 of 13 MSI-H patients showed ACVR2A mutation. Our results indicate a strong association between ACVR2A mutation and MSI-H in gastric cancer patients.

Perineural invasion (PN) diagnosed by hematoxylin-eosin (HE) staining is an important prognostic factor after curative-intent surgery in patients with colorectal cancer. However, the clinical significance of PN diagnosed by immunohistochemistry (IHC) has not been investigated. The present study assessed the clinical significance of PN diagnosed by IHC with an anti-S100 antibody in patients with colorectal cancer.We retrospectively enrolled 184 consecutive patients with stage I-III colorectal cancer who had undergone curative-intent surgery. We analyzed the absence/presence of PN diagnosed by HE staining (HE-PN) compared to that diagnosed by IHC with the anti-S100 antibody (S100-PN). Potential prognostic factors were identified by univariate and multivariate analyses of the overall and relapse-free survival. The [Formula: see text] statistics were used to assess the inter-observer reproducibility.The incidence of HE-PN and S100-PN among the 184 patients was 60 patients (32.6%) and 113 patients (61.4%), respectively (P0.001). A multivariate Cox proportional hazards regression model analysis indicated that S100-PN was an independent prognostic factor for both the overall and relapse-free survival. The [Formula: see text] value was 0.77 for S100-PN and 0.47 for HE-PN.PN diagnosed by IHC is an important prognostic factor in patients with colorectal cancer. An inter-observer assessment showed superior judgment reproducibility for S100-PN compared with HE-PN.

Breast cancer drug development costs nearly $610 million and 37 months in preclinical mouse model trials with minimal success rates. Despite these inefficiencies, there are still no consensus breast cancer preclinical models. Murine mammary adenocarcinoma 4T1-luc2 cells were implanted subcutaneous (SQ) or orthotopically percutaneous (OP) injection in the area of the nipple, or surgically into the chest 2nd mammary fat pad under direct vision (ODV) in Balb/c immunocompetent mice. Tumor progression was followed by in vivo bioluminescence and direct measurements, pathology and survival determined, and tumor gene expression analyzed by genome-wide microarrays. ODV produced less variable-sized tumors and was a reliable method of implantation. ODV implantation into the chest 2nd mammary pad rather than into the abdominal 4th mammary pad, the most common implantation site, better mimicked human breast cancer progression pattern, which correlated with bioluminescent tumor burden and survival. Compared to SQ, ODV produced tumors that differentially expressed genes whose interaction networks are of importance in cancer research. qPCR validation of 10 specific target genes of interest in ongoing clinical trials demonstrated significant differences in expression. ODV implantation into the chest 2nd mammary pad provides the most reliable model that mimics human breast cancer compared from subcutaneous implantation that produces tumors with different genome expression profiles of clinical significance. Increased understanding of the limitations of the different preclinical models in use will help guide new investigations and may improve the efficiency of breast cancer drug development .

Tuberculosis (TB) and HIV are considered pandemic by the World Health Organization (WHO). It has been reported that HIV infection is one of the major risk factors for the development of TB, increasing the incidence by up to 1,000 times, but it often has an atypical presentation. The incidence of extrapulmonary TB is increasing, largely among HIV patients. The diagnosis of intestinal TB is a challenge because of its chronic and nonspecific presentation which often mimics other diseases, and requires a high clinical suspicion to timely diagnose. Massive lower gastrointestinal bleeding due to intestinal TB was once an uncommon complication of TB, but recent reports indicate an increased incidence especially in developing countries. We suspect that co-infection with cytomegalovirus colitis contributes to the massive hemorrhage from intestinal TB. Surgical intervention is the recommended management for intestinal TB complicated by lower gastrointestinal bleeding. Accordingly, it is important for HIV patients to be screened and treated for TB to prevent this complication. Although the diagnosis is a challenge, it is important to consider intestinal TB as a cause of gastrointestinal bleeding in the HIV positive patients.

UNLABELLED Bile acids are important hormones during the feed/fast cycle, allowing the liver to coordinately regulate nutrient metabolism. How they accomplish this has not been fully elucidated. Conjugated bile acids activate both the ERK1/2 and AKT signaling pathways via sphingosine 1-phosphate receptor 2 (S1PR2) in rodent hepatocytes and in vivo. Here, we report that feeding mice a high-fat diet, infusion of taurocholate into the chronic bile fistula rat, or overexpression of the gene encoding S1PR2 in mouse hepatocytes significantly upregulated hepatic sphingosine kinase 2 (SphK2) but not SphK1. Key genes encoding nuclear receptors/enzymes involved in nutrient metabolism were significantly downregulated in livers of S1PR2(-/-) and SphK2(-/-) mice. In contrast, overexpression of the gene encoding S1PR2 in primary mouse hepatocytes differentially increased SphK2, but not SphK1, and mRNA levels of key genes involved in nutrient metabolism. Nuclear levels of sphingosine-1-phosphate, an endogenous inhibitor of histone deacetylases 1 and 2, as well as the acetylation of histones H3K9, H4K5, and H2BK12 were significantly decreased in hepatocytes prepared from S1PR2(-/-) and SphK2(-/-) mice. CONCLUSION Both S1PR2(-/-) and SphK2(-/-) mice rapidly developed fatty livers on a high-fat diet, suggesting the importance of conjugated bile acids, S1PR2, and SphK2 in regulating hepatic lipid metabolism.

Sphingosine-1-phosphate (S1P) is a pleiotropic lipid mediator that regulates cell survival, migration, the recruitment of immune cells, angiogenesis, and lymphangiogenesis, all of which are involved in cancer progression. S1P is generated inside cancer cells by sphingosine kinases then exported outside of the cell into the tumor microenvironment where it binds to any of five G protein coupled receptors and proceeds to regulate a variety of functions. We have recently reported on the mechanisms underlying the “inside-out” signaling of S1P, its export through the plasma membrane, and its interaction with cell surface receptors. Membrane lipids, including S1P, do not spontaneously exchange through lipid bilayers since the polar head groups do not readily go through the hydrophobic interior of the plasma membrane. Instead, specific transporter proteins exist on the membrane to exchange these lipids. This review summarizes what is known regarding S1P transport through the cell membrane via ATP-binding cassette transporters and the spinster 2 transporter and discusses the roles for these transporters in cancer and in the tumor microenvironment. Based on our research and the emerging understanding of the role of S1P signaling in cancer and in the tumor microenvironment, S1P transporters and S1P signaling hold promise as new therapeutic targets for cancer drug development.

Sphingosine-1-phosphate (S1P), a pleiotropic bioactive lipid mediator, and the kinase that produces it have now emerged as key regulators of numerous cellular processes involved in inflammation and cancer. Here, we review the importance of S1P in colitis and colitis-associated cancer (CAC) and discuss our recent work demonstrating that S1P produced by upregulation of SphK1 during colitis and associated cancer is essential for production of the multifunctional NF-κB-regulated cytokine IL-6, persistent activation of the transcription factor Stat3, and consequent upregulation of the S1P receptor, S1PR1. The effectiveness of the pro-drug FTY720 (known as fingolimod), approved for the treatment of multiple sclerosis, has become the gold standard for S1P-centric drugs, and will be used to illustrate the therapeutic value of modulating SphK1 and S1P receptor functions. We will discuss our recent results showing that FTY720/fingolimod administration interferes with the SphK1/S1P/S1PR1 axis and suppresses the NF-κB/IL-6/Stat3 malicious amplification loop and CAC. These preclinical studies suggest that FTY720/fingolimod may be useful in treating colon cancer in individuals with ulcerative colitis.

TWO MOST COMMONLY USED ANIMAL MODELS FOR STUDYING BREAST CANCER LUNG METASTASIS ARE: lung metastasis after orthotopic implantation of cells into the mammary gland, and lung implantations produced after tail vein (TV) injection of cells. Tail vein injection can produce lung lesions faster, but little has been studied regarding the differences between these tumors, thus, we examined their morphology and gene expression profiles.Syngeneic murine mammary adenocarcinoma, 4T1-luc2 cells, were implanted either subcutaneously (Sq), orthotopically (OS), or injected via TV in Balb/c mice. Genome-wide microarray analyses of cultured 4T1 cells, Sq tumor, OS tumor, lung metastases after OS (LMet), and lung tumors after TV (TVt) were performed 10 days after implantation.Bioluminescence analysis demonstrated different morphology of metastases between LMet and TVt, confirmed by histology. Gene expression profile of cells were significantly different from tumors, OS, Sq, TVt or LMet (10,350 probe sets; FDR≤1%; P0.0001). Sq tumors were significantly different than OS tumors (700 probe sets; FDR≤15%; P0.01), and both tumor types (Sq and OS) were significantly different than LMet (1,247 probe sets;1.5-fold-change; P0.01), with no significant difference between TVt and LMet.There were significant differences between the gene profiles of cells in culture and OS versus LMet, but there were no differences between LMet versus TVt. Therefore, the lung tumor generated by TVt can be considered genetically similar to those produced after OS, and thus TVt is a relevant model for breast cancer lung metastasis.

Fifteen per cent of all patients who undergo surgery for colorectal cancer require palliation for obstruction or perforation, which usually entails a fecal diverting loop colostomy.1 The reported risk of peristomal hernia for all patients with colostomy ranges from 5 to 50 per cent and prolapse ranges from 2 to 22 per cent.1 Management of these complications in the palliative setting must take into consideration the diminished life expectancy of the patient with advanced metastatic disease, the impact of a morbid recovery in the setting of reduced longevity, and the degree to which an operation may reduce an already limited quality of life.2 Patients with advanced colorectal cancer have advanced peritoneal disease, malnutrition, immune compromise, chronic pain, and elevated abdominal and/or portal pressures and are thus at increased risk of morbidity, recurrence, and postsurgical complications. Accordingly, management of ostomy prolapse and peristomal hernias in this patient population has usually focused on palliative medical symptom management rather than surgery, except in situations of obstruction or strangulation.1 This report describes a technique we used to perform colostomy revision using a minimally invasive approach in a patient with a large prolapse and concomitant peristomal hernia that incorporated the small bowel. This was in the setting of a patient with advanced rectal cancer and provided significant symptom relief without the morbidity of a laparotomy. The patient was a 56-year-old woman who previously had a diverting loop colostomy performed as a palliative procedure for an advanced obstructing and perforated adenocarcinoma. In addition to a 20-cm prolapse of the proximal end of her loop colostomy, she had peritoneal carcinomatosis and a computed tomography scan at that time demonstrated persistent hepatic metastases, suspicious pulmonary nodules, a pericardial effusion, and thrombosis of the portal, superior mesenteric and splenic veins with varices and ascites. The large stomal prolapse contained an adherent sliding peristomal hernia of a loop of jejunum that did not appear to be obstructed or strangulated. She had no peritoneal signs on physical examination and no leukocytosis. She required admission to the hospital for pain control and we were consulted for consideration of colostomy revision. Because of her advanced disease and high risk of morbidity from an open operation, we undertook a minimally invasive approach to the palliation of her symptoms. After general anesthesia was induced, the prolapsed colostomy was prepped and draped, and the distal orifice was closed temporarily with a running silk suture to limit fecal contamination during the procedure. The mucosal surface of the prolapsed limb of the loop colostomy was incised using cautery to expose the contents of the prolapse at a point opposite to the herniated jejunal loop as shown in Figure 1. This full-thickness incision provided exposure of the contents of the hernia without laparotomy, including the mesentery, colonic walls, small bowel, and adhesive bands. Blunt and sharp dissection was used to safely take down the adhesions and free up the small bowel, which were then easily reduced through this minimally invasive approach. Fig. 1 The schematic demonstration of exposing the contents of the prolapse through a full-thickness incision made in the mucosa contralateral to the hernia through the serosa. Using eight staple loads of the 63.8-mm GIA stapler, the colostomy was revised as follows. Accessing the stomal lumen through the previously closed orifice of the prolapsed limb, while simultaneously confirming no hernia into the prolapse through the exposure of the incision by direct vision and palpation, the stapler was advanced longitudinally down the length of prolapsed limb toward the base to a level of 2 cm above the abdominal wall (Fig. 2). Then staples were fired in a transverse fashion at the level of the 2-cm base along the circumference of the prolapsed stoma to maintain that new 2-cm height of the base above the abdominal wall while simultaneously confirming under direct vision that no structures would be injured. The colostomy was then matured in the usual fashion. The double-barrel ostomy was pink, viable, patent, and productive without obstruction, prolapse, or hernia. The patient had an uncomplicated postoperative course. Her presenting symptom of debilitating abdominal pain improved remarkably. She was discharged after a five-day course of prophylactic antibiotics with an immediate resumption of her pre-operative diet and it achieved high patient satisfaction. The patient died five months later as a result of cancer progression without prolapse recurrence. Fig. 2 The stapler was applied down the prolapsed limb and then across the base 2 cm above the abdominal wall with direct visualization of the reduced hernia contents. The reports in the literature of palliating colostomy prolapse without laparotomy in terminal cancer patients describe blind stapling of the prolapsed segment.1–4 These authors all reported palliation without complication but had limited follow-up as a result of patients dying of their disease. Our approach with full-thickness mucosal through a serosal incision to provide transluminal exposure of the contents of the prolapse before stapling provides the benefit of palliation without the morbidity of laparotomy, but it does this by providing greater exposure and control than the blind stapling methods. It should be noted that there are several interesting points to consider. First is whether five days of postoperative antibiotics to prevent infectious complications is open to question with a valid argument for only pre-operative antimicrobial prophylaxis. Second, the use of a pursestring suture to control fecal contamination4 provides more efficient access to the lumen than the running closure we describe. Third, the potential criticism that this approach is likely to produce a high recurrence rate if the patient survives long enough is valid because we do not repair the defect, although it did palliate the patient’s symptoms with minimal morbidity. Although use of mesh to prevent recurrence may be considered, we believe the risks of infection are too high through a transluminal approach in this high-risk patient population. There are prospective randomized trials underway investigating the role of biologic mesh to prevent and treat peristomal hernias. Although this minimally invasive approach does not replace laparotomy in patients being treated with curative intent, it does have a role in palliating symptoms.

e13125Background: Human epidermal growth receptor type 2 (HER2) overexpression/amplification is associated with the efficacy of HER2 targeted therapies in breast and gastric cancers, where candidates for its targeted therapies are selected by immunohistochemistry (IHC) and/or fluorescence in situ hybridization (FISH) testing. However, the HER2 status in colorectal cancer (CRC) has not been fully investigated to date. CancerPlex is a comprehensive next-generation sequencing-based test that evaluates 400-plus genes for mutations and copy number changes, including HER2. In the present study, we aimed to clarify the association between HER2 status and HER2 gene alteration detected with CancerPlex in CRC. Methods: This retrospective study included 100 patients with Stage I-IV CRC, who were evaluated HER2 status and HER2 gene alteration. IHC for HER2 was classified as 4 grades according to the immunoreactivity. The patients with IHC2+ were further tested with FISH. IHC3+/IHC2+ and FISH positive patients were de...

e15121Background: The advent of whole genomic sequencing has greatly facilitated the identification and characterization of cancer-relevant genes. Given the potential genetic and environmental diff...

e15103Background: It has been previously documented that high somatic mutational rates in cancer as determined by whole genome sequencing correlates with high microsatellite instability (MSI-H) and...

Inflammation is a central process in several disorders and contributes to cancer progression. Inflammation involves a complex cascade of pro-inflammatory and anti-inflammatory signaling events with protein and lipid mediators. Recent advances in lipid detection have revealed the importance of lipid mediators in inflammation. Omega three polyunsaturated fatty acids (ω-3 PUFA) are found naturally in fish oil and have been extensively studied in multiple inflammatory diseases with improved outcomes. Resolvins are thought to be the active metabolites of ω-3 PUFA, and are responsible for facilitating the resolving phase of acute inflammation. Clinically, resolvins have been associated with resolution of acute kidney injury and acute lung injury, micro and macro vascular response to injury, and inhibition of microglia-activated inflammation in neurodegenerative disorders. In addition to inflammatory diseases, ω-3 PUFA and resolvins appear to modulate cancer progression. ω-3 PUFA intake has been associated with reduced inflammation in colorectal cancer, and favorable phenotype in breast cancer. Resolvins offer promising therapeutic potential as they may modulate inflammation with minimal side-effects, in contrast to currently available anti-inflammatory medications. This review describes the roles of ω-3 PUFA and resolvins in the inflammatory cascade, various inflammatory diseases, and specific cancers. Additionally, it will discuss the clinical therapeutic potential of resolvins as targets in inflammatory diseases and cancers.

Meckel’s diverticulum is a common asymptomatic congenital gastrointestinal anomaly, but rarely it can present with hemorrhage. Over the last few years inverted Meckel’s diverticulum has been reported in the literature with increasing frequency as an occult source of lower gastrointestinal hemorrhage. Here, we report a case of a 54-year-old male, who was referred for surgical evaluation with persistent anemia and occult blood per rectum after a work up which failed to localize the source over 12 mo, including upper and capsule endoscopy, colonoscopy, enteroclysis, Meckel scan, and tagged nuclear red blood cell scan. An abdominal computed tomography scan showed a possible mid-ileal intussusception and intraluminal mass. During the abdominal exploration, inverted Meckel’s diverticulum was diagnosed and resected. We review the literature, discuss the forms in which the disease presents, the diagnostic modalities utilized, pathological findings, and treatment. Although less than 40 cases have been reported in the English literature from 1978 to 2005, 19 cases have been reported in the last 6 years alone (2006-2012) due to improved diagnostic modalities. Successful diagnosis and treatment of this disease requires a high index of clinical suspicion, which is becoming increasingly relevant to general gastroenterologists.

Sphingosine-1-phosphate (S1P) is a potent sphingolipid metabolite that regulates a number of biological processes critical for cancer. S1P produced inside cancer cells is exported and exerts its extracellular functions by binding to its specific receptors in an autocrine, paracrine, and/or endocrine manner, which is known as inside-out signaling. S1P is also known to exert its intracellular functions especially in the inflammatory process, but its relevance to cancer biology remains to be elucidated. Recently, there have been growing interests in the role of S1P in breast cancer progression, including angiogenesis and lymphangiogenesis. Our group demonstrated that activation of sphingosine kinase 1, the enzyme that catalyzes the phosphorylation of sphingosine to S1P, is a key step of this process. In this review, we will cover our current knowledge on the role of S1P signaling pathway in breast cancer progression with an emphasis on its role in tumor-induced lymphangiogenesis.

Background Although many retrospective studies suggest that resection of the primary tumor improves survival in metastatic breast cancer, animal studies suggest that resection induces metastasis. Moreover, there has been no critical evaluation of how well animal studies actually model metastatic breast cancer. We used our newly established orthotopic cancer implantation under direct vision model to evaluate the hypothesis that primary tumor resection improves survival in metastatic breast cancer by reducing overall tumor burden and improving immune responsiveness. Methods Murine mammary adenocarcinoma 4T1-luc2 cells that can be visualized by bioluminescence were implanted orthotopically into BALB/c mice under direct vision. Resection of the primary tumors at days 6, 10, and 28 were compared to sham resection of the contralateral normal mammary gland and observation alone. Tumor burden was quantified by bioluminescence. Tumor-draining lymph nodes were identified by intradermal injection of lymphazurin, and primary tumors, lymph nodes, and lungs were examined pathologically. Kaplan-Meier survival analyses were performed. Splenocyte myeloid-derived suppressor cells (MDSCs) and CD4 or CD8 single positive T lymphocytes were quantified by flow cytometry. Results Tumors invaded locally, metastasized to regional lymph nodes, and then metastasized to distant organs, with subsequent mortality. Surgical stress increased tumor burden only transiently without affecting survival. When primary tumor resection decreased overall tumor burden substantially, further growth of metastatic lesions did not increase the overall tumor burden compared to observation, and survival was improved, which was not the case when resection did not significantly reduce the overall tumor burden. Decreasing overall tumor burden through resection of the primary tumor resulted in decreased splenic MDSC numbers and increased CD4 and CD8 cells, suggesting the potential for an improved immunologic response to cancer. Conclusion Decreasing overall tumor burden through resection of the primary breast tumor decreased MDSCs, increased CD4 and CD8 cells, and improved survival.

Sphingosine-1-phosphate (S1P) is a pleiotropic bioactive lipid mediator that promotes breast cancer progression by diverse mechanisms that remain somewhat unclear. Here we report pharmacologic evidence of a critical role for sphingosine kinase 1 (SphK1) in producing S1P and mediating tumor-induced hemangiogenesis and lymphangiogenesis in a murine model of breast cancer metastasis. S1P levels increased both in the tumor and the circulation. In agreement, serum S1P levels were significantly elevated in stage IIIA human breast cancer patients, compared with age/ethnicity-matched healthy volunteers. However, treatment with the specific SphK1 inhibitor SK1-I suppressed S1P levels, reduced metastases to lymph nodes and lungs, and decreased overall tumor burden of our murine model. Both S1P and angiopoietin 2 (Ang2) stimulated hemangiogenesis and lymphangiogenesis in vitro, whereas SK1-I inhibited each process. We quantified both processes in vivo from the same specimen by combining directed in vivo angiogenesis assays with fluorescence-activated cell sorting, thereby confirming the results obtained in vitro. Notably, SK1-I decreased both processes not only at the primary tumor but also in lymph nodes, with peritumoral lymphatic vessel density reduced in SK1-I–treated animals. Taken together, our findings show that SphK1-produced S1P is a crucial mediator of breast cancer–induced hemangiogenesis and lymphangiogenesis. Our results implicate SphK1 along with S1P as therapeutic targets in breast cancer. Cancer Res; 72(3); 726–35. ©2012 AACR.

P53-binding protein 1 (53BP1) is an early DNA damage response-protein that is rapidly recruited to sites of DNA double-strand breaks. The presence of 53BP1 nuclear foci can be considered as a cytologic marker for endogenous double-strand breaks reflecting genomic instability. This study aimed to clarify the early DNA damage response mediated by 53BP1 in tumor specimens of ductal resection margins and to elucidate its predictive value for clinically evident local recurrence at ductal stumps in 110 patients undergoing resection for extrahepatic cholangiocarcinoma. The ductal resection margin status was classified as negative (85 patients), positive with carcinoma in situ (14 patients), or positive with invasive carcinoma (11 patients). The nuclear staining pattern of 53BP1 was evaluated by immunofluorescence. TUNEL analysis was used to calculate apoptotic index. Ductal margin status was the only independent risk factor for local recurrence (P=0.001). The cumulative probability of local recurrence at 5 years was 10%, 40% and 100% in patients with negative ductal margins, positive with carcinoma in situ and positive with invasive carcinoma, respectively (P

Sphingosine 1-phosphate (S1P), a potent sphingolipid mediator produced by sphingosine kinase isoenzymes (SphK1 and SphK2), regulates diverse cellular processes important for breast cancer progression acting in an autocrine and/or paracrine manner. Here we show that SphK1, but not SphK2, increased S1P export from MCF-7 cells. Whereas for both estradiol (E(2)) and epidermal growth factor-activated SphK1 and production of S1P, only E(2) stimulated rapid release of S1P and dihydro-S1P from MCF-7 cells. E(2)-induced S1P and dihydro-S1P export required estrogen receptor-alpha, not GPR30, and was suppressed either by pharmacological inhibitors or gene silencing of ABCC1 (multidrug resistant protein 1) or ABCG2 (breast cancer resistance protein). Inhibiting these transporters also blocked E(2)-induced activation of ERK1/2, indicating that E(2) activates ERK via downstream signaling of S1P. Taken together, our findings suggest that E(2)-induced export of S1P mediated by ABCC1 and ABCG2 transporters and consequent activation of S1P receptors may contribute to nongenomic signaling of E(2) important for breast cancer pathophysiology.