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Table S1. Clinical Characteristics of multiple cohorts Table S2. Association of MSI-H subtypes with clinical and genomic characteristics in multiple cohorts Table S3. Association of dMMR subtypes with clinical characteristics in Cohort 4 Supplementary Figure S1. Prognostic significance of MSI/MMR status in patients with colorectal cancer. Supplementary Figure S2. Prognostic significance of TGFB expression in patients with MSI-H colorectal cancer. Supplementary Figure S3. Expression z-scores for genes encoding immune checkpoint molecules for MSI-H-Stromal, MSI-H-Conventional and MSS tumors in Cohort 1. Supplementary Figure S4. Correlation of TGFβ-stromal signature (TBSS) z-score with stage of disease in Cohort 2 (A) and Cohort 3 (B). Supplementary Figure S5. Expression z-scores for genes encoding transcription factors that regulate Th cell differentiation, including TBX21 (Th1), GATA3 (Th2) and RORC (Th17) for MSI-H-Stromal, MSI-H-Conventional and MSS tumors in Cohort 1 (A), Cohort 2 (B) and Cohort 3 (C). Supplementary Figure S6. Genomic aberrations in 43 TGFβ pathway genes, including TGFβ superfamily ligands, receptors and receptor-associated proteins, SMADs, and adaptor molecules. Supplementary Figure S7. Correlation between RNA-seq-based transcriptomic TBSS and mass spectrometry-based proteomic TBSS in the CPTAC dataset.

Patients with DNA mismatch repair–deficient (dMMR)/microsatellite instability-high (MSI-H) colorectal cancer represent a biomarker-defined population with distinct clinicopathologic features who are susceptible to immune checkpoint inhibitors (ICI). However, their survival outcomes vary considerably and nearly half of them exhibit primary resistance to current ICIs, suggesting substantial molecular heterogeneity even among tumors with dMMR/MSI-H. We conducted an extensive analysis of the tumor microenvironment (TME) using multiple transcriptomic, proteomic, and IHC cohorts of colorectal cancer, comprising 222 dMMR/MSI-H and 1440 MMR-proficient/microsatellite stable tumors. We developed a TGFβ-responsive stromal gene signature and then identified a unique poor prognostic subgroup of patients with dMMR/MSI-H colorectal cancers, characterized by the upregulation of transcriptional programs, including the TGFβ-rich active TME, angiogenesis, M2 macrophage polarization, and the extracellular matrix signature predictive of ICI resistance. The TGFβ-dependent stromal subset within dMMR/MSI-H tumors exhibiting poor survival outcomes was further recapitulated by proteomic datasets and IHC for VCAN protein expressed by cancer-associated fibroblasts. Meanwhile, this dMMR/MSI-H stromal subgroup was enriched neither with CD8+ T-cell infiltration nor common genomic alterations, such as mutation density and BRAF mutations, compared with dMMR/MSI-H tumors without TGFβ-dependent stromal activation. In conclusion, this study revealed a novel stromal subgroup of patients with dMMR/MSI-H colorectal cancer, demonstrating a TGFβ-rich tumor-promoting TME and unfavorable survival outcomes.Implications:Dual inhibition of immune checkpoints and TGFβ signaling may offer a promising strategy for these patients.

Univariate and multivariate analyses of risk factors for OS and RFS after NAC in patients with ESCC

S1. Measurement of the number of CD163+ cells and their expression intensities in ESCC specimens. S2. IHC staining of CSF-1, IL-34, E-cadherin, and vimentin in ESCC specimens. S3. Correlation between mRNA expression of CD163 and CSF-1 in patients with ESCC who underwent curative esophagectomy without any preoperative therapy. S4. The effect of CSF-1 expression of NAC responsivity and tumor progression after NAC in ESCC. S5. Involvement of higher fold-change of IL-34 expression after CDDP treatment to poorer chemo-sensitivity in ESCC cells. S6. Enhanced mRNA expression of IL-34 in CDDP-resistant human ESCC cells.

The tumor microenvironment (TME) plays a key role in the efficacy of neoadjuvant chemotherapy (NAC) in solid tumors including esophageal squamous cell carcinoma (ESCC). However, the TME profile of ESCC treated with NAC is not fully understood. In this study, we investigated the effect of NAC on the TME especially tumor-associated macrophages (TAM), the important immunosuppressive components of the TME, in ESCC. We quantified the expression of CD163, a crucial marker of TAM, in pretherapeutic biopsy and surgically resected ESCC specimens from patients who received NAC (n = 33) or did not receive NAC (n = 12). We found that NAC dramatically increased the expression of CD163 on TAMs in ESCC. Colony-stimulating factor 1 (CSF-1) and IL34 are crucial cytokines that recruit monocytes into tumor sites and differentiate them into TAMs. Interestingly, NAC significantly upregulated the expression of IL34 but not CSF-1 on tumor cells, and the frequencies of CD163+ TAMs were significantly correlated with IL34 expression in ESCC after NAC. The expression of IL34 in NAC-nonresponsive patients was significantly higher than that in NAC-responsive patients, and patients with IL34-high ESCC exhibited worse prognosis as compared with patients with IL34-low ESCC. We also demonstrated that 5-fluorouracil (5-FU)/cisplatin preferentially increased mRNA expression of IL34 on human ESCC cell lines. Human peripheral blood monocytes co-cultured with ESCC cells treated with 5-FU/cisplatin increased the expression of CD163, which was attenuated by the treatment with CSF-1R inhibitors. These data suggest that IL34 expression by NAC shifts the TME toward CD163+ TAM-rich immunosuppressive and chemo-insensitive microenvironment in ESCC.Implications:The blockade of IL34 signaling may offer a novel therapeutic strategy against chemoresistance in ESCC by inhibiting M2-TAM polarization.

The tumor microenvironment (TME) plays a key role in the efficacy of neoadjuvant chemotherapy (NAC) in solid tumors including esophageal squamous cell carcinoma (ESCC). However, the TME profile of ESCC treated with NAC is not fully understood. In this study, we investigated the effect of NAC on the TME especially tumor-associated macrophages (TAM), the important immunosuppressive components of the TME, in ESCC. We quantified the expression of CD163, a crucial marker of TAM, in pretherapeutic biopsy and surgically resected ESCC specimens from patients who received NAC (n = 33) or did not receive NAC (n = 12). We found that NAC dramatically increased the expression of CD163 on TAMs in ESCC. Colony-stimulating factor 1 (CSF-1) and IL34 are crucial cytokines that recruit monocytes into tumor sites and differentiate them into TAMs. Interestingly, NAC significantly upregulated the expression of IL34 but not CSF-1 on tumor cells, and the frequencies of CD163+ TAMs were significantly correlated with IL34 expression in ESCC after NAC. The expression of IL34 in NAC-nonresponsive patients was significantly higher than that in NAC-responsive patients, and patients with IL34-high ESCC exhibited worse prognosis as compared with patients with IL34-low ESCC. We also demonstrated that 5-fluorouracil (5-FU)/cisplatin preferentially increased mRNA expression of IL34 on human ESCC cell lines. Human peripheral blood monocytes co-cultured with ESCC cells treated with 5-FU/cisplatin increased the expression of CD163, which was attenuated by the treatment with CSF-1R inhibitors. These data suggest that IL34 expression by NAC shifts the TME toward CD163+ TAM-rich immunosuppressive and chemo-insensitive microenvironment in ESCC. Implications: The blockade of IL34 signaling may offer a novel therapeutic strategy against chemoresistance in ESCC by inhibiting M2-TAM polarization.

Epstein-Barr virus-positive gastric cancer [EBV (+) GC] is a distinct GC subtype with unique genetic and epigenetic aberrations. Here, we examined resected GC samples and publicly available microarray data and The Cancer Genome Atlas (TCGA) database to identify the mechanism underlying overexpression of PD-L1 in EBV (+) GC. We found that high levels of PD-L1 overexpression in EBV (+) GC were caused by focal amplification of CD274. By contrast, relatively high expression of PD-L1 in tumor tissue and infiltrating immune cells correlated with CD8 lymphocyte infiltration and IFN-γ expression via IRF3 activation. Since we reported previously that PD-L1 expression is associated both with the presence of CD8 T cells in the tumor microenvironment and with IFN-γ expression in GC, we examined a database to see whether IFN-γ-associated overexpression of PD-L1 plays a significant role in EBV (+) GC. Immunohistochemical staining showed that expression of the IRF3 signature in clinical GC samples was higher in EBV (+) than in EBV (−) cases. The data presented herein reveal a unique dual mechanism underlying PD-L1 overexpression in EBV (+) GC: high focal amplification of CD274 or IFN-γ-mediated signaling via activation of IRF3.

BackgroundSystemic inflammation has been reported to be associated with cancer progression and metastasis. Systemic inflammation score (SIS), calculated from preoperative serum albumin level and lymphocyte-to-monocyte ratio, has been shown to be a novel prognostic factor for several types of tumors. This study aimed to evaluate the prognostic value of the SIS in patients with pT2-4 resectable gastric cancer (GC).MethodsTotal 97 patients with pT2-4 GC who underwent curative surgery between 2009 and 2015 in Fukushima Medical University Hospital were included. We performed univariate and multivariate analyses to evaluate the usefulness of preoperative SIS and other prognostic factors for relapse-free survival (RFS) and overall survival (OS).ResultsThe higher SIS score was associated with undifferentiated cancer and recurrence. Univariate analysis of RFS identified deeper tumor invasion and higher SIS were significant risk factors and multivariate analysis revealed that both of them were independent prognostic factors for RFS. As for OS, age, tumor invasion, SIS and LNR were significantly correlated with RFS. In multivariate analysis, tumor invasion, SIS and LNR were independent prognostic factors for OS.ConclusionsSIS was an independent prognostic factor for RFS and OS in pT2-4 resectable gastric cancer patients who underwent curative gastrectomy.

AT-rich interactive domain 1A (ARID1A), which is a tumor suppressor gene, is frequently mutated in Epstein-Barr virus-positive gastric cancer [EBV (+) GC]. While most ARID1A mutations in GC are truncating mutations, leading to loss of ARID1A protein expression, epigenetic modifications appear to contribute to ARID1A deficiency in EBV (+) GC harboring wild-type ARID1A. Based on the significant role of epigenetic modifications in EBV (+) GC that contributes to ARID1A deficiency, the methylation status of ARID1A was evaluated in EBV-infected cells and GC patients using a publicly available microarray and the Cancer Genome Atlas (TCGA) database. EBV-encoded miRNAs that potentially target ARID1A were identified as an additional epigenetic modulator by computational prediction. In vitro experiments were conducted to evaluate how EBV-encoded miRNAs affected ARID1A mRNA and protein levels. In clinical GC samples, the expression of predicted miRNAs and ARID1A and the mutation status of ARID1A was evaluated. As results, ARID1A was not hypermethylated in EBV (+) GC samples or EBV-infected GC cells. EBV infection did not alter ARID1A mRNA levels, suggesting that ARID1A protein deficiency was caused by post-transcriptional gene silencing in ARID1A-WT EBV (+) GC. Overexpression of miR-BART11-3p and miR-BART12, which were identified as miRNAs that potentially bind ARID1A, suppressed ARID1A protein expression in MKN7 and NCI-N87 cells. Highly expressed miR-BART11-3p and miR-BART12 were correlated with decreased ARID1A levels in GC tumors which did not harbor ARID1A mutations. The present findings revealed that ARID1A expression was epigenetically regulated by miR-BART11-3p and miR-BART12 in EBV (+) GC.

Highlights • A rare case of Tuberculosis Peritonitis regarding the aspect of perioperative infection control. • The importance of characteristic intraperitoneal findings, which contributes to the rapid and accurate diagnosis combined with pathological findings. • Diagnostic laparoscopy must be less invasive and more effective compared with conventional methods., Background Tuberculous peritonitis (TBP) is uncommon in Japan, and its diagnosis with conventional methods is time taking and requires a high clinical index of suspicion. Laparoscopy with peritoneal biopsy is a tool for rapid and accurate diagnosis of TBP. However, few cases have mentioned the infectious control and prevention during the perioperative period. This case is written following the SCARE scale for case report writing. Case presentation A 30-year-old man from Southeast Asia with a past medical history of pulmonary tuberculosis at 3-year-old admitted to our institution with abdominal pain and slight fever lasting for a week. With the elevation of inflammatory response and CA125, we conducted CT (Computed tomography). Not only ascites, panniculitis with peritoneal nodules, and the thickening of the omentum were found. Considering the possibility of malignancy and TBP, we performed a diagnostic laparoscopy. Slightly cloudy ascites, peritoneal and thickening omentum with white nodules were seen, and pathological diagnosis from the omentum during the operation raised the possibility of TBP due to its caseating granuloma and these findings allowed us to start the rapid treatment. Conclusion We reported the effectiveness of diagnostic laparoscopy along with the aspect of perioperative prevention for TBP.

The patient was a 66-year-old male who had undergone an operation for lung cancer and solitary brain metastases. Follow- up PET-CT after 1 year detected FDG accumulation in the stomach. We performed esophagogastroscopy and found an approximately 20 mm-sized Type 2 tumor on the greater curvature of the upper stomach. A pathological diagnosis of lung adenocarcinoma metastasis in the stomach was made. Laparoscopic surgery was performed on the metastatic lesion to prevent bleeding and perforation, and resection was achieved with minimal invasion. The current development of chemotherapy, including immunotherapy, has contributed to the improved prognosis of cancer patients, including those with lung metastasis in the stomach. Considering these backgrounds, preventive surgical resection under laparoscopy may be an effective approach for improving prognosis and preventing acute life-threatening adverse events. We report this case along with a literature review.

Objectives Left colic artery preserving lymph node dissection around the inferior mesenteric artery (IMA) is a standard procedure for rectal cancer surgery. Although the IMA sheath is a well-known structure, to our knowledge, there are no reports describing its microanatomy from an oncological point of view; therefore, there is no consensus on how to handle the sheath for accurate lymph node dissection around IMA. We aimed to investigate the components of the IMA sheath pathologically, focusing particularly on the presence of lymph nodes (LNs) and lymphatic ducts (LDs). Methods We evaluated rectal and sigmoid cancer specimens resected with high-tie technique in our institute in April 2017-April 2018. The specimens were collected consecutively, without any selection. In the resected specimens, the entire anatomical structure of IMA was investigated. We defined the IMA sheath as the tissues located between the surface of the IMA adventitia and collagenous layers connecting the outermost nerve fibers. The microanatomy around the IMA was examined using H&E staining, and LDs were identified using D2-40 immunohistochemistry. Results Twenty patients were enrolled. No LNs were observed within the sheath in any of the cases. However, there were a significant number of LDs (11.08 ± 3.35) within the sheath. Conclusions Our anatomical definition of IMA sheath was feasible and objectively possible. These microanatomical results partially support the surgical concept of left colic artery preserving lymph node dissection around the IMA. It may be difficult to remove all lymphatic ducts without removing the IMA itself.

A small subset of patients with proficient mismatch repair (pMMR)/microsatellite stable (MSS)-colorectal cancer (CRC) benefit from immunotherapy with anti-programmed cell death 1 (PD-1)/programmed death ligand 1 (PD-L1) blockade. Therefore, the aim of the current study was to evaluate the immune status of patients with pMMR/microsatellite instability-low (MSI-L)/MSS-CRC and deficient MMR (dMMR)/MSI-high (MSI-H)-CRC in order to identify responders to anti-PD-1/PD-L1 inhibitors. The current study used a dataset downloaded from The Cancer Genome Atlas (TCGA) as well as 219 clinical tissue samples to investigate the infiltrating grade of cluster of differentiation (CD) 4 and CD8 tumor infiltrating lymphocytes (TILs), the expression levels of PD-L1 and PD-L2, the interferon-γ (IFN-γ) and CD8 T effector gene signatures, and the phosphorylated signal transducer and activator of transcription 1 (p-STAT1) status in patients with pMMR/MSI-L/MSS-CRC and dMMR/MSI-H-CRC. Analysis of TCGA dataset revealed that the mRNA expression levels of PD-L1 and PD-L2, the IFN-γ gene signature and the CD8 T effector gene signature were significantly upregulated in MSI-H tumors compared with MSI-L/MSS tumors. Additionally, a subpopulation of patients with upregulation of the IFN-γ and CD8 T effector gene signatures was observed in those with MSI-L/MSS-CRC. Immunohistochemical staining of the clinical samples revealed a subpopulation of patients with pMMR-CRC that were positive for PD-L1 and p-STAT1, and whom had levels of elevated CD8(+) and CD4(+) TILs infiltration similar to those observed in patients with dMMR-CRC. The results obtained in the current study suggested that a subpopulation of patients with MSI-L/MSS-CRC and pMMR-CRC with upregulated IFN-γ and CD8 T effector gene signatures may benefit from immunotherapy with antibodies against PD-1 and PD-L1.

INTRODUCTION: Recently in Japan, Ramucirumab (RAM) became the first anti-angiogenic agent to be approved for second-line treatment of gastric cancer. In the present study, we aimed to evaluate the efficacy and safety of RAM plus paclitaxel (PTX) in patients with unresectable and recurrent gastric cancer in our institution. PATIENTS AND METHODS: The subjects were 11 patients with unresectable and recurrent gastric cancer who received RAM plus PTX as a second- or later-line treatment at our hospital between June 2015 and September 2017, after the failure of previously-attempted treatments. We assessed the efficacy and safety of RAM plus PTX, and also compared them between patients aged

Background Upper extremity deep vein thrombosis (UEDVT) is relatively rare but cannot be negligible because it can cause fatal complications. Although it is reported that the occurrence rate of UEDVT has increased due to central venous catheter (CVC), cancer, and surgical invasion, there are still limited information for esophagectomy. The aim of this study was to evaluate the clinical factors, including CVC placement and thromboprophylaxis approach, as well as retrosternal space’s width as a predictive factor for UEDVT in patients receiving esophagectomy. Methods This study included 66 patients who underwent esophagectomy with retrosternal reconstruction using a gastric tube. All patients routinely underwent contrast-enhanced computed tomography (CT) on the 4th postoperative day. Low-molecular-weight-heparin (LMWH) was routinely administered by the 2nd postoperative day. To evaluate retrosternal space’s width, (a) The distance from sternum to brachiocephalic artery and (b) the distance from sternum to vertebra were measured by preoperative CT, and the ratio of (a) to (b) was defined as the width of retrosternal space. Results Among all patients, 11(16.7%) suffered from UEDVT, and none was preoperatively received CVC placement, while 7 were inserted in non-UEDVT cases. Retrosternal space’s width in patients with UEDVT was significantly smaller than that in patients without UEDVT (0.17 vs. 0.26; P < 0.0001). A cutoff value of the width was 0.21, which has high sensitivity (87%) and specificity (82%) for UEDVT prediction, respectively. Conclusion The existence of CVC may not affect the development of UEDVT, but preoperative evaluation of retrosternal ratio may predict the occurrence of UEDVT.

Cancer-associated fibroblasts (CAFs) in the tumor microenvironment play an essential role in the tumor progression of esophageal squamous cell carcinoma (ESCC). The present study aimed to investigate the expression of CAF-related molecules, versican, periostin and lumican, in cancer stroma, to provide prognostic stratification for patients with ESCC after surgery. A total of 106 patients with ESCC who underwent curative esophagectomy without preoperative chemotherapy or radiotherapy were enrolled. The expression of CAF-related stromal proteins, including versican, periostin and lumican, was examined using immunohistochemistry, and the prognostic value was assessed by Kaplan-Meier survival analysis, and univariate and multivariate Cox regression models. The expression of versican, periostin and lumican was found specifically in the stromal component of ESCC. Kaplan-Meier analysis demonstrated that, compared with a low expression level, a high expression level of versican, periostin or lumican in the cancer stroma was significantly associated with worse relapse-free survival (RFS) and overall survival times in patients with ESCC. The prognostic values of stromal versican and lumican remained significant in a stratified analysis of stage I patients. Moreover, univariate and multivariate analysis revealed that high stromal versican or lumican expression was an independent prognostic factor for RFS in the patients. The present study demonstrated that CAF-related molecules, including versican, periostin and lumican, were expressed in the stroma of ESCC, and that stromal expression of versican and lumican in particular may have clinical utility as a prognostic biomarker for poor RFS in postoperative patients with ESCC.

Surgical resection is the most effective treatment for liposarcoma, a retroperitoneal malignant soft tissue tumor, and a reliable negative margin is required because of the high risk of local recurrence. We reported a case of pelvic liposarcoma that could be resected by laparoscopic and transsacral hybrid approach. A 60's-man had a mixed liposarcoma occupying the right rear of the pelvis in the rectum. The operation was preceded by a laparoscopic operation, and the right internal iliac artery and vein and the superior rectal artery were dissected. The tumor was separated along the right pelvic wall. The oral rectum was transected and the colon was elevated by the extraperitoneal route. After conversion to the Jackknife position, the anterior sacrum was exfoliated with the right transsacral approach, the coccyx was resected, and the rectal anus, tumor, and surrounding fatty tissue were removed as an en bloc fasion. Histopathological examination showed mixed type of liposarcoma and negative margin of the stump. The patient is alive without recurrence 8 months after the surgery.

Patients with DNA mismatch repair–deficient (dMMR)/microsatellite instability-high (MSI-H) colorectal cancer represent a biomarker-defined population with distinct clinicopathologic features who are susceptible to immune checkpoint inhibitors (ICI). However, their survival outcomes vary considerably and nearly half of them exhibit primary resistance to current ICIs, suggesting substantial molecular heterogeneity even among tumors with dMMR/MSI-H. We conducted an extensive analysis of the tumor microenvironment (TME) using multiple transcriptomic, proteomic, and IHC cohorts of colorectal cancer, comprising 222 dMMR/MSI-H and 1440 MMR-proficient/microsatellite stable tumors. We developed a TGFβ-responsive stromal gene signature and then identified a unique poor prognostic subgroup of patients with dMMR/MSI-H colorectal cancers, characterized by the upregulation of transcriptional programs, including the TGFβ-rich active TME, angiogenesis, M2 macrophage polarization, and the extracellular matrix signature predictive of ICI resistance. The TGFβ-dependent stromal subset within dMMR/MSI-H tumors exhibiting poor survival outcomes was further recapitulated by proteomic datasets and IHC for VCAN protein expressed by cancer-associated fibroblasts. Meanwhile, this dMMR/MSI-H stromal subgroup was enriched neither with CD8+ T-cell infiltration nor common genomic alterations, such as mutation density and BRAF mutations, compared with dMMR/MSI-H tumors without TGFβ-dependent stromal activation. In conclusion, this study revealed a novel stromal subgroup of patients with dMMR/MSI-H colorectal cancer, demonstrating a TGFβ-rich tumor-promoting TME and unfavorable survival outcomes. Implications: Dual inhibition of immune checkpoints and TGFβ signaling may offer a promising strategy for these patients.

AT-rich interactive domain 1A (ARID1A) is a tumor suppressor gene that is frequently mutated in gastric cancer (GC). Although ARID1A mutations are not a druggable target for conventional treatments, novel therapeutic strategies based on a synthetic lethal approach are effective for ARID1A-deficient cancers. The histone methyltransferase EZH2 acts in a synthetic lethal manner in ARID1A-mutated ovarian cancer, although its role in GC remains unknown. The selective sensitivity of the EZH2 inhibitors for ARID1A-deficient GC cells was evaluated using cell viability and colony formation assays. The expression of PI3K/AKT signaling genes were investigated using TCGA’s cBioPortal database to determine whether the homeostasis between ARID1A and EZH2 is related to cell proliferation and survival via the PI3K/AKT signaling pathway. We also evaluated the phosphorylation of PI3K/AKT signaling proteins in ARID1A knock downed ARID1A-WT GC cells. EZH2 inhibitors decreased the viability of ARID1A-deficient cells in a dose-dependent manner and demonstrated the selective sensitivity to ARID1A-deficient cells in vitro experiment system. Bioinformatics approach revealed that the PI3K/AKT signaling was tended to be activated in ARID1A-deficient GC enhancing cell viability and, furthermore, down-regulation of EZH2 in ARID1A-deficient GC was related to normalization of PI3K/AKT signaling pathway. The cell experiment revealed that phosphorylated AKT was upregulated in ARID1A-deficent GC cells. The present findings provide a rationale for the selective sensitivity of EZH2 inhibitors against ARID1A-deficient GC and suggest the potential efficacy of targeted therapy using EZH2 inhibitors in this patient population.

Cancer vaccines and immune checkpoint inhibitors (ICI) have recently been employed as immunotherapies for esophageal squamous cell carcinoma (ESCC). Cancer vaccines for ESCC have yielded several promising results from investigator-initiated phase I and II clinical trials. Furthermore, a Randomized Controlled Trial as an adjuvant setting after curative surgery is in progress in Japan. On the other hand, ICI, anti-CTLA-4 mAb and anti-PD-1 mAb, have demonstrated tumor shrinkage and improved overall survival in patients with multiple cancer types. For ESCC, several clinical trials using anti-PD-1/anti-PD-L1 mAb are underway with several recent promising results. In this review, cancer vaccines and ICI are discussed as novel therapeutic strategies for ESCC.

Adjuvant chemotherapy is considered for patients with stage II colorectal cancer (CRC) characterized by poor prognostic clinicopathological features; however, current stratification algorithms remain inadequate for identifying high-risk patients. To develop prognostic assays, we conducted a step-wise screening and validation strategy using nine cohorts of stage II patients based on multiple platforms, including microarray, RNA-sequencing (RNA-seq) and immunohistochemistry (IHC) on formalin-fixed paraffin-embedded (FFPE) tissues. Four microarray datasets (total n = 458) were used as the discovery set to screen for single genes associated with postoperative recurrence. Prognostic values of candidate genes were evaluated in three independent microarray/RNA-seq validation cohorts (n = 89, n = 93 and n = 183, respectively), and then IHC for KRT17 was conducted in two independent FFPE series (n = 110 and n = 44, respectively). We found that high levels of KRT17 transcript expression were significantly associated with poor relapse-free survival (RFS) not only in the discovery set, but also in three validation cohorts, and its prognostic impact was independent of conventional factors by multivariate analyses. Positive staining of KRT17 protein was significantly associated with poor RFS in two independent FFPE cohorts. KRT17 protein expression had independent prognostic impact on RFS in a multivariate model adjusted for conventional variables, including high-risk clinicopathological features. In conclusion, using nine independent cohorts consisting of 997 stage II patients, we identified and validated the expression of KRT17 transcript and KRT17 protein as a robust prognostic biomarker that can discriminate postoperative stage II patients who are at high probability of disease recurrence, providing additional prognostic stratification beyond the currently available high-risk factors.

In the original publication of the article, Fig. 1 was published incorrectly.

Our previous differential transcriptome analysis between a paired specimen of normal and esophageal squamous cell carcinoma (ESCC) tissues found aberrant expression of kallikrein-related peptidase 13 (KLK13) in tumors. In this study, we evaluated the expression of KLK13 in many ESCC cases in relation with clinical features, and the prognosis. Eighty-eight ESCC cases were subjected to immunohistological staining for KLK13 and classified into KLK13-negative and KLK13-positive groups. Difference of clinical features and the prognosis between the groups was analyzed. In normal esophageal mucosa, KLK13 expression was evident but limited in the stratum granulosum in all cases. By contrast, only 27 of 88 ESCC samples showed KLK13 expression, whereas the remaining 61 tumors showed no KLK13 expression. The KLK13-positive group was significantly associated with pT classification (deeper tumor invasions; P = 0.0282), pN classification (lymph node metastasis; P = 0.0163), and advanced TNM stage (P = 0.0198). In KLK13-positive samples, KLK13-expressing cells often expressed Ki67, a proliferation marker, unlike normal mucosa, in which Ki67-expressing cells were limited to the basal layer and did not express KLK13. Compared with patients with KLK13-negative group, KLK13-positive group showed poorer postoperative prognosis. Relatively high levels of KLK13 expression in ESCC were associated with cell proliferation and correlated with tumor progression, advanced cancer stage, and poor prognosis.

Background Recently, endoscopic submucosal dissection (ESD) has been used as a less invasive treatment for superficial esophageal cancer. Additional treatment is often required after non-curative resection to prevent local recurrence and lymph node metastasis. Here, we present the outcomes of various additional treatments for patients with superficial esophageal cancer who underwent ESD. Methods Between 2006 and 2017, we performed ESD in 179 patients (210 lesions) with superficial esophageal cancer and 44 cases resulted in the non-curative resection diagnosed by the pathological examination. Among them, 29 patients received additional treatment, whereas 15 patients with no additional treatment were followed up. Additional treatment included esophagectomy (8 patients), chemoradiotherapy (15 patients), ablation using argon plasma coagulation (4 patients), and chemotherapy alone (2 patients). We examined the clinicopathological characteristics and prognosis of patients in the additional esophagectomy group (S group) and chemoradiotherapy group (CRT group). Results Twenty-three patients with pT1a-MM, pT1b, lymphatic invasion, venous invasion, and positive resection margins (both horizontal and vertical) were divided into two treatment groups. Clinicopathological characteristics of patients in the S and CRT groups were not significantly different. Pathological findings after additional esophagectomy showed one residual tumor and one lymph node metastasis. There were no recurrences in the two groups. There was no statistically significant difference in the 5-year overall survival rate between the S group (87.5%) and the CRT group (93.3%). One patient from the S group died due to respiratory pneumonia, and one patient died due to radiation pneumonia. However, five out of the 15 (33.3%) patients who were followed up with no additional treatment developed recurrence. The 5-year overall survival rate was 40.4%, which was not significantly different from that in the additional treatment group. However, the 5-year relapse-free survival rate (30%) was significantly different from that in the additional treatment group (P > 0.05). Conclusion Additional treatment is essential after non-curative endoscopic submucosal resection for esophageal cancer. Additional esophagectomy and chemoradiotherapy were both safe and effective in this cohort. Disclosure All authors have declared no conflicts of interest.

Background Neuroendocrine cell carcinoma (NEC) of the esophagogastric junction is rare and usually has a very poor prognosis. Methods Here we present two cases of NEC occurred in the esophagogastric junction. Results Case 1 A 50-year-old man was admitted to the introduction origin medical institute with an abdominal pain and dysphagia. Upper gastrointestinal endoscopy revealed a type 2 tumor at the esophagogastric junction, and the pathological examination showed the diffuse proliferation of relatively homogeneous tumor cell with chromatin-enriched nuclear and immunohistologically, the tumor cells were positive for Chromogranin A, CD56, AE1/3. MIB-1 index was 80%, we diagnosed neuroendocrine carcinoma (small cell type). TNM Stage was GE, Type 3, cT4, cN1, cM0 cStage IIIB (ENETS TNM classification) He had undergone total gastrectomy and lower esophagectomy with transhiatal approach and 2 field of lymph node dissection. Pathological examination revealed NEC component developed under the muscularis mucosa, differentiated adenocarcinoma localized upper the muscularis mucosa and Chromogranin A positive cells were scattered inside. Pathological findings showed NEC (MIB-1 72.5%) with tub1, 70 × 56 mm, pT3 pN1(7/36), stage IIIB (HER2 score0). Adjuvant chemotherapy using S-1 was started, but the follow up CT showed recurrence in mediastinum, left subclavian and paraaortic lymph nodes 7 months after surgery. S-1 followed by CPT-11 + CDDP, CT showed the shrinkage of paraaortic lymph nodes metastasis. The patient alive for 55 months without any evidence ofprogression being continued chemotherapy. Case 2 A 57-year-old man was admitted to the introduction origin medical institute with dysphagia. Upper gastrointestinal endoscopy revealed a type 2 tumor at the esophagogastric junction, and the pathological examination showed NEC (small cell type). CT and PET revealed mediastinal lymph node metastasis, aortic invasion and adrenal metastasis. TNM stage was NEC, EG, cT3, cN1, cM1 cStage IV. We performed a systemic chemotherapy with CPT-11 + CDDP, the evaluation of treatment effect after 5 course chemotherapy revealed partial response. However the patient underwent the endoscopic stent graft due to stenosis, and died due to progressive disease 18months after chemotherapy induction. Conclusion We reported here two cases of NEC occurring in the esophagogastric junction. It's clinical behavior remains unclear and the treatment strategy for NEC of esophagogastric junction is not established. Further investigation of accumulated cases of this rare entity is necessary. Disclosure All authors have declared no conflicts of interest.

Background Circulating tumor cells (CTCs) in patients with malignant tumors can be used as a prognostic marker. Recently the existence of mesenchymal CTCs have been detected by new methods. However, it has been not yet clarified how CTCs are associated with the treatment effects in esophageal cancer patients. We assessed CTCs in esophageal cancer patients and investigated the relationship between CTCs and treatment effect. Methods Seven patients who had potential of curative resection have been enrolled and peripheral blood samples (10ml) were collected before and after treatment. All patients received chemotherapy (5-FU and cisplatin) and four patients of them received as neoadjuvant therapy. Other patients received only chemotherapy and radiation therapy without operation. CTCs were analyzed using a Microfluidic Chip devise provide with the Nihon Gene Research Laboratories. This system can isolates CTCs from blood samples, based on their size and deformability differences from blood cells. Phenotypes of CTCs are determined by staining and scanning systems. A previous report revealed that this method had higher sensitivity for CTCs than conventional methods (the Cell Search system) in 61 metastatic breast cancer. Results Treatments effects were stable disease or better in all cases. Multiple CTCs were detected in all cases before treatment. Five patients had epithelial CTCs and others had only mesenchymal CTCs. Total number of CTCs after treatment whose data can be available decreased except for two cases. One patient had no CTC after treatment. Two cases have been currently analyzing. Conclusion CTCs may exist almost all patients of Stage II or more esophageal cancer. Decreasing the number of CTCs after treatment suggests some relationship between CTCs and treatment effect, and the accumulation of more cases is necessary. Disclosure All authors have declared no conflicts of interest.