- Email: [email protected]
New Quantum BCH Codes of Length n=2(q4−1)
treatment were examined retrospectively. Patients who received no oral administration of BCAA were included as a control group (C group), while those who orally received one dose of Aminoleban EN powder mix for three days after the endoscopic treatment were included as a BCAA group. Endpoints (patient backgrounds and blood tests) were examined by: intergroup comparison at one week and at one month after the endoscopic treatment and temporal changes within each group. Those aged 65 years or older were examined as described above. The Institutional Review Board of Osaka Medical College approved this study. (No. 2312) [Results] The C group included 9 cases, while the BCAA group included 16 cases. The mean ages were 69 and 65 years in the C and BCAA groups, respectively. The sex ratios (male: female) were 7: 2 and 9: 7 in the C and BCAA groups, respectively. Child-Pugh classifications (A: B: C) were 7: 2: 0 and 8: 7: 1 in the C and BCAA groups, respectively. There was no significant difference in characteristics of patients between the two groups. The intergroup comparison demonstrated a significantly higher total peripheral blood lymphocyte count (TLC) in the BCAA group than in the C group at one week after the endoscopic treatment. Similar results were obtained in those aged 65 years or older. The temporal changes showed no significant difference in the C group between the start and one week after the endoscopic treatment. However, the GPT, ALP, γ-GPT, and Glu levels were significantly decreased in the BCAA group at one week after the treatment. The intergroup comparison demonstrated a significantly higher TLC in the BCAA group than in the C group at one month after the treatment. Similar results were obtained in those aged 65 years or older. The temporal changes showed no significant difference in the C group. However, the GPT and Glu levels were significantly decreased in the BCAA group at one week but returned to the baseline levels at one month. [Conclusion] Immune and liver functions and glucose tolerance are improved by combining branched-chain amino acidenriched enteral nutrition during the endoscopic treatment of esophageal varices. However, an effective administration method should be considered for elderly patients.
Su2028 NATURAL HISTORY OF PERCUTANEOUS ENTERAL GASTROSTOMY TUBE IN PATIENTS WITH AMYOTROPHIC LATERAL SCLEROSIS Mohanad T. Al-Qaisi, Richard Gerkin, Logan Cobb, Mary Chew, Katherine Petersen, Rakesh Nanda Background: Amyotrophic lateral sclerosis (ALS) is a devastating disease that puts a grave burden on patients' quality of life. It is associated with rapid weight loss due to dysphagia and increased metabolism, which eventually leads to early mortality. Percutaneous endoscopic gastrostomy (PEG) is usually performed for nutritional support for a variety of reasons. Studies that looked at effects of PEG tubes on mortality are limited. Aim: We aimed to examine the baseline characteristics, functional status and overall survival for patients with ALS. Methods: Retrospective cohort study patients diagnosed with ALS at a large VA specialty ALS clinic in a metropolitan city between 01/2010 and 08/2017 with at least three months of follow up. Patients' demographics (age, gender, baseline functional status, weight and BMI at time of diagnosis,) were gathered. Use of riluzole, weight and BMI at 1 year were ascertained using chart review on follow up visits at the ALS clinic. Survival analysis was calculated using a Cox regression model. A p-value of 0.05 was considered statistically significant. Results: 97 consecutive patients were included in the study. 52 (53%) patients received PEG tube during study time. Baseline characteristics are in table 1. Overall, our patients lived a median of 750 days from diagnosis (95% CI 465-1034). Interestingly, PEG patients had lower median survival (730 days from diagnosis (95% CI 593-867)) compared to non-PEG patients (909 days from diagnosis (95% CI 409-1409), however this did not reach statistical significance (p = 0.70). PEG patients were younger and had lower median baseline BMI (24.3 kg/m2(+/-4) compared to non-PEG patients (28.1 kg/m2(+/-5). After 1 year, PEG patients' median BMI dropped to 22.5 kg/m2(+/-5), still less than non-PEG patients (26.9 kg/m2(+/-6). Conclusion: PEG placement does not significantly affect patient's survival in patients with ALS. More studies are required to help understand the effects of PEG tube placement in patients with ALS.
Age yrs (SD) Gender Male N (%) Baseline BMI kg/m2 (SD) Baseline weight kg (SD) BMI after 1 year from diagnosis kg/ m2 (SD) Weight after 1 year from diagnosis kg (SD) Median follow up days (SD) Baseline functional status: Wheelchair N (%) Riluzole N (%) Aspiration N (%)
PEG (n=52) 65.0 (10.0) 49 (94.2)
No PEG (n=47) 69.8 (7.5) 44 (97.8)
p-Value 0.03 0.62
24.3 (4)
28.1 (5)
0.39
74.8 (13.6)
90.2 (17.0)
0.39
22.5 (5)
26.9 (6)
0.46
69.6 (19.7)
87.5 (19.3)
0.40
667 (674)
455 (460)
0.36
36 (78.3)
21 (60.0)
0.08
34 (68.0) 29 (76.3)
31 (68.9) 4 (15.4)
0.93 <0.0001
Su2039 USE OF PATIENT-DERIVED TUMOR XENOGRAFT MODEL TO GUIDE TREATMENT DECISIONS FOR PATIENTS Yanping Zhu, Chun-Mei Zhao, Yanpeng Hao, Zhiqiang Wang, Li Wang, Duan Chen Background/aim: Patient-derived tumor xenograft (PDTX) models are established by direct engraftment of patient-derived tumor fragments into immunocompromised mice. Approx. 20, 000 PDTXs have been so far created in academic and industry labs in US and Europe for basic and preclinical cancer research, such as biomarker discovery, drug screening for personalized medicine, understanding of drug-resistance mechanisms and novel therapy development. The aim of our work was to use PDTXs to guide clinical decisions for patients. Materials and Methods: From January 2015 to November 2018 at 5 clinical centers in China, we have collected tumor samples by surgery, biopsy and/or endoscopy from 723 patients with esophageal cancer (12 cases), gastric cancer (84), colorectal cancer (65), liver cancer (294), pancreatic cancer (46), sarcomas (94), gynecological cancer (63), lung cancer (26), and others (34). In 434 out of 723 PDTXs, the primary tumors (P0) grew to 800 mm3 in mice, and then re-implanted into mice to produce the first passage (P1). When the P1 tumors grew to 100–150 mm3, clinical tests of treatment regimens were conducted. It took 3-5 months from tumor sampling to clinical report. The cost was about 200 000 RMB (approx. 30,000 USD) per patient. In the remaining 289 PDTX, the P0 tumors grew in a “tumor-microenvironment matrix” that was implanted in mice, named as superPDTX (patent application no. 201810916601.5). Biomarkers of superPDTX included Ki67 index and pathological scores. It took 2 weeks at a cost of 30 000 RMB (or 5 000 USD) per patients. In addition, whole exome sequencing (WES) was performed in 8 superPDTXs. Results: The overall engraftment rate of PDTX (P1) was 56.2%, e.g. esophageal cancer was 45.5%, gastric cancer was 63.9%, colorectal cancer was 62.2%, liver cancer was 53.8%, and pancreatic cancer was 52.6%. A cohort study of 21 PDTXs/patients showed that the consistency of treatment effects was 85.7% (18/21) in which 15 out of 17 was validated in co-clinical trial and 3 out of 4 was in predictive testing. A total of 289 superPDTXs were used for pharmaceutical drug screening (61 cases) and clinical testing (228 patients, including 160 GI cancers). The successful rate of superPDTX (P0) clinical testing was 92%. Another cohort of 22 superPDTXs/patients receiving 28 treatment regimens showed a consistency rate of 82.1% (23/28) in which 8 out of 13 was validated in co-clinical trial and 15 out of 15 was in predictive testing. Up-to-date, we have banked the PDTX tissue samples (P0-P2) of > 35 000. WES data showed that >90% mutations identified in primary tumors were present in the corresponding superPDTX samples (P0). Conclusion: Based on our results and clinical practice, we suggested that superPDTX testing could be used routinely to enhance clinical decision making for personalized cancer care. Su2040 DIVERSE TUMOR SUSCEPTIBILITY IN COLLABORATIVE CROSS MICE: IDENTIFICATION OF A NEW MODEL FOR HUMAN GASTRIC TUMORIGENESIS Pin Wang, susan Celniker, Xiaoping Zou, Bo Hang, Antoine M Snijders, Jianhua Mao Objective: The Collaborative Cross (CC) is a mouse population model with diverse and reproducible genetic backgrounds used to identify novel disease models and genes that contribute to human disease. Since spontaneous tumor susceptibility in CC mice remains unexplored, we assessed tumor incidence and spectrum. Design: We monitored 293 mice from 18 CC strains for tumor development. Genetic association analysis and RNA-sequencing were used to identify susceptibility loci and candidate genes. We analyzed genomes of gastric cancer patients to evaluate the relevance of genes identified in the CC mouse model and measured the expression levels of ISG15 by immunohistochemical staining using a gastric adenocarcinoma tissue microarray. Association of gene expression with overall survival (OS) was assessed by Kaplan-Meier analysis. Results: CC mice displayed a wide range in the incidence and types of spontaneous tumors. More than 40% of CC036 mice developed gastric tumors within one year. Genetic association analysis identified Nfκb1 as a candidate susceptibility gene, while RNA-sequencing analysis of non-tumor gastric tissues from CC036 mice showed significantly higher expression of inflammatory response genes. In human gastric cancers the majority of human orthologs of the 166 mouse genes were preferentially
Su2029 USEFULLNESS OF BRANCHED-CHAIN AMINO ACID-ENRICHED ENTERAL NUTRITION IN THE ENDOSCOPIC TREATMENT OF ESOPHAGEAL VARICES Yuichi Kojima, Toshihisa Takeuchi, Shinya Nishida, Noriaki Sugawara, Kazuhiro Ota, Satoshi Harada, Kazuhide Higuchi [Background] Fasting during the endoscopic treatment of esophageal varices may deteriorate the nutritional statuses of patients with cirrhosis, suggesting the need for nutritional intervention to maintain the hepatic functional reserve. [Objective] To examine the usefulness of branched-chain amino acid-enriched enteral nutrition in the endoscopic treatment of esophageal varices. [Methods] Of patients who underwent endoscopic treatment for esophageal varices in Osaka Medical College Hospital between January 2014 and June 2017, those who did not take a branched-chain amino acid (BCAA) preparation at the beginning of the
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in 1 patient (7.7%). We recorded baseline characteristics such as age, sex, comorbidities, indications for PTEG, and preoperative biomarkers. We evaluated clinical outcomes, such as adverse events, postoperative length of stay, place of discharge, readmission rates, and mortality. Results: Of the 13 patients, 10 underwent PTEG for receiving nutrition and 3 underwent PTEG to reduce the pressure on the intestinal tract. Underlying conditions that required PTEG were gastric resection in 10 patients (76.9%), left diaphragm disorder in 2 patients (15.4%), and an interposing transverse colon or liver between the abdominal wall and anterior gastric wall in 1 patient (7.7%). PTEG was successful in all patients. The mean procedure time of PTEG was 47.9±22.3 minutes. As a postoperative adverse event, 1 patient (7.7%) dislodged the tube accidentally and required reinsertion of a new tube. No mortality was observed at 30 days. The mean postoperative length of stay was 48.5 (12–69) days. Two patients (15.4%) were discharged home, and 8 (61.5%) were discharged to long-term care hospitals. Three patients (23.1%) died of primary diseases before discharge; 1 patient (7.7%) was able to receive full nutrition orally, and this patient's percutaneous transesophageal gastric tube was dislodged on postoperative day 81. Conclusions PTEG is an effective and safe method to provide nutrition or reduce the pressure of the intestinal tract. PTEG may be an alternative technique when gastrostomy is impossible.
AGA Abstracts
altered by amplification or deletion and were significantly associated with OS. Higher expression of the CC036 inflammatory response gene signature is associated with poor OS. Finally, ISG15 protein is elevated in gastric adenocarcinomas, and correlated with shortened patient OS. Conclusions: CC strains exhibit tremendous variation in tumor susceptibility, and we present CC036 as a spontaneous laboratory mouse model for studying human gastric tumorigenesis.
Su2044 EXOSOMES RELEASED FROM PATIENT-DERIVED ORGANOIDS CONTAIN DIFFERENT MICRORNA PROFILES BETWEEN COLORECTAL CANCER AND ADENOMA Hiroshi Nagai, Masatake Kuroha, Atsushi Masamune Exosomes are membrane-enclosed nanovesicles and have been increasingly recognized as important cell communications by transmitting microRNA (miRNA). No previous studies have addressed the exosomal miRNA profile in colorectal adenoma (CRA) due to the difficulty of long-term culture of CRA. Development of a three-dimensional organoid culture system allows us to perform long-term expansion of epithelial organoids of human CRA and CRC. The aim of this study was to clarify the differences in exosomal and cellular miRNA profiles between organoids derived from CRA and CRC. We established six CRC patient-derived organoids and eight CRA patient-derived organoids. Exosomes were prepared from conditioned medium of patient-derived organoids. We analyzed the exosomal and cellular microRNA expression profiles by miRNA microarray. We found different miRNA expression profiles between CRC- and CRA-derived organoids, and that both exosomal and cellular miR-1246 expression was upregulated in CRC-derived organoids compared to CRA-derived organoids. Alteration of miR-1246 expression by miR-1246 mimic or inhibitor accordingly increased or decreased cell proliferation in HT-29 CRC cell line, respectively. In conclusion, we here reported the first study dissecting the miRNA profile in exosomes in CRA- and CRC-derived organoids. Upregulation of miR-1246 might play a role in increased cell proliferation in colorectal adenoma-carcinoma transition.
Su2042 DERIVATION OF OESOPHAGEAL ORGANOIDS TO RECAPITULATE THE HETEROGENEITY OF PRIMARY TUMOURS AND PROVIDE A MODEL SYSTEM FOR PRECISION THERAPEUTICS Xiaodun Li, Hayley Francies, Ahmad Miremadi, Amber Grantham, Nicola Grehan, Ginny Devonshire, Mathew Garnett, Rebecca C. Fitzgerald Oesophageal adenocarcinoma (EAC), the dominant subtype of oesophageal cancer in western countries, has increased six-fold in the last 30 years. Despite concerted efforts to improve patient management the overall 5-year survival is still below 15% due to late diagnosis and limited response to traditional therapy. Whole genome sequencing efforts of the ICGC have enabled us to better characterise the genomic landscape which is dominated by copy number changes and structural rearrangements with enormous heterogeneity in point mutations (SNVs) and InDels. The genomic heterogeneity has made it difficult to ascribe rational targeted therapy. However, the more homogeneous mutational signatures and integrated analysis of -omits and expression changes suggest new therapeutic hypotheses. In order to test these hypotheses model systems are required. To overcome the drawback of historical 2D lines with missing germline status and unreliable EAC mouse models, we have developed primary EAC organoids with matched WGS data for the primary tumour and germline. To maxim the power of cellular model to recapitulate this extreme heterogeneous cancer, to date 27 organoids have been developed with a lifespan over 6 months, which provides the biggest EAC cell model collection so far. These organoids are representative of the clinical disease: 68% of organoids are from male patients with an average age of 67 years; 18 of 27 patients received chemo treatment and12 of these chemo treated patients showing chemo resistance (Tumour Regression Grade score > 3). Based on our recent description of the genomic landscape of 551 EAC patients, 74 of 76 driver mutations could also be found in this organoid cohort. For example, 78% of harbour TP53mutations, 33% ARID1A, and 29% SMAD4. A thorough phenotypic and molecular characterisation has confirmed that the organoids faithfully recapitulate the parental tumour at a genomic protein level and have enabled us to study clonality, cell kinetics and tumour polarity. Organoid models in combination with genomic characterisation of the matched primary provides a functional platform to develop rational therapeutic choices in EAC.
Su2045 VASCULAR SHUT DOWN EFFECTS OF PHOTODYNAMIC THERAPY WITH TALAPORFIN Taketo Suzuki, Mamoru Tanaka, Hiroshi Ichikawa, Hirotada Nishie, Hiromi Kataoka Background: Photodynamic therapy (PDT) is an attractive modality for cancer therapy because of minimally invasive and minimally toxic. PDT consists of the intravenous administration of a photosensitizer, followed by activation with a specific wavelength light. Activation of the photosensitizer causes the conversion of molecular oxygen into various reactive oxygen species that directly induce the death of the tumor cells or shut down the tumor-associated vasculature. However, the mechanism of the vascular shut down is not clear. Here, we evaluated that PDT with Talaporfin (mono-l-aspartyl chlorin6, Laserphyrin®) shut down existing tumor vessels. Materials and Methods: (1) Talaporfin was administered to human umbilical vein endothelial cell lines (HUVEC) followed by irradiation with a 664 nm LED. The cell death-inducing effects were analyzed by WST-8 assay and half maximal inhibitory concentration (IC50) was calculated. (2) Tubulin and F-actin (actin stress fiber) were analyzed by immunofluorescent staining in HUVEC treated with PDT. The phosphorylation of myosin light chain (MLC) was examined by Western blotting. And we evaluated the relationship with Rho-GTP pathway by the Rho-kinase inhibitor; Y-27632. (3) Xenograft tumor mouse models were established with colon cancer cell lines (HCT116). After Talaporfin intravenous injection, tumors were illuminated with the diode laser system (664nm). Tumor blood flow was measured by a laser speckle blood flow analyzing system; OMEGA ZONE2. Temporal changes of the flow were compared with the positive control; Combretastatin A-4 phosphate (CA4P). Result: (1) PDT with Talaporfin induced the cell death effectively in HUVEC. (IC50 = 13.6 cˇmol/l). (2) PDT induced the depolymerization of tubulin and the polymerization of F-actin because of destruction of the microtubule. But the Rho-kinase inhibitor Y-27632 prevented the polymerization of F-actin. It indicated that PDT induced the vascular shut down effect by polymerization of F-actin via the Rho-GTP pathway. (3) PDT with Talaporfin decreased the blood flow in tumors but not in the neighboring healthy tissue. And the blood flow rate significantly decreased compared with CA4P. Conclusions: PDT with Talaporfin induced vascular shut down effect by activating of Rho-GTP pathway leading to F-actin polymerization in vitro and decreased the tumor blood flow significantly in vivo. In summary, we could show the vascular shut down of PDT with Talaporfin which suggested as an important part of its antitumor effects.
Su2043 A NOVEL CANINE INTESTINAL ORGANOID MODEL TO CHARACTERIZE DOSE-EXPOSURE-RESPONSE OF CHEMOTHERAPEUTIC DRUGS Dana C. Borcherding, Yoko M. Ambrosini, Todd Atherly, Samantha Thomson, Larry W. Wulf, David Borts, Albert Jergens, Karin Allenspach, Jonathan P. Mochel Background: Stem cell-derived 3D organoids have emerged as a cutting-edge cell culture technology for drug discovery, toxicity screening, and testing for personalized medicine. Given that gastro-intestinal (GI) toxicity is one of the commonly reported side effects of chemotherapeutics, it is important to evaluate the dose-exposure-GI toxicity response relationship of anticancer agents in a scalable in vitro system prior to their use in patients. To bridge the knowledge gap between rodents and humans, our lab has developed a three dimensional (3D) canine GI organoid (enteroid and colonoid) system for testing of chemotherapeutic drugs. Large animal models, such as the dog, are indeed more representative of humans than mice as they have a relatively large body size, longer life span, more closely resemble human GI physiology and develop spontaneous, analogous diseases including colorectal cancer (CRC). The objective of this proof-of-concept study was to demonstrate how canine organoids can be used to establish drug dose-exposure-toxicity relationships using Doxorubicin as a testbed. Methods: Canine 3D intestinal organoids cultured in matrigel were exposed to control or Doxorubicin for 2 days before matrigel was dissolved by Matrisperse. Doxorubicin concentration in organoids was then determined by liquid chromatography mass spectrometry (LC-MS) and compared to a 6-point standard curve (50 to ~14,000 pg range) to determine Doxorubicin concentration within organoid samples. The toxicity of Doxorubicin on 3D canine organoids in matrigel matrix was determined after 48 hours of incubation with Doxorubicin by MTT assay. Results: were compared to the cytotoxicity of Doxorubicin (1-1000 nM) in two CRC cell-lines, HCT-116 and HT-29, by MTT assay, with data evaluated by ANOVA and t-tests. Results: Exposure of canine organoids to Doxorubicin was confirmed by LC-MS, with uptake of about 10% or ~34 ng per 12 wells of organoids after incubation with 100 nM Doxorubicin for 2 days. We observed a mild, dose-dependent cytotoxicity of ~15-25% by 100-1000 nM Doxorubicin (p<0.05) after 2 days in small intestinal (jejunum) organoids from a healthy dog by the MTT assay. In comparison, Doxorubicin significantly (p<0.05) reduced cell viability in two CRC cell-lines (HCT-116 and HT-29) to about 40-60% and 10-20% of control for 100 and 1000 nM Doxorubicin, respectively. Conclusions: We confirmed that Doxorubicin was functional, and, as expected, exerted only a mild cytotoxic effect on the non-cancer intestinal organoids but showed much higher cytotoxicity on the fast proliferating CRC cells. These studies establish proof-of-concept that canine intestinal organoids can be used for in vitro doseexposure-cytotoxicity characterization of chemotherapeutic drugs.
AGA Abstracts
Su2046 MORPHOLOGIC DIFFERENCE OF PATIENTS-DERIVED IMMORTAL ESOPHAGEAL CANCER ORGANOIDS WAS ASSOCIATED WITH MOLECULAR AND POST-GENOMIC DIFFERENCE Su Youn Nam, Hee Jeong Lim, Su In Kim Background: Primary organoid have advantages in providing more physiologically relevant and predictive data for in-vivo response. Previously, we showed high agreement between clinical response and response in patents-derived primary organoid. We produced immortal patient derived esophageal cancer organoid and investigated the morphologic and molecular characteristics supporting the correlation between primary tumor and organoid. Method: Patient-derived primary organoid culture was performed using tumor tissues acquired from esophageal squamous cell cancer. We cultured them up to 1000um diameter and then dissected into 3-4 organoids. Next-generation sequencing (NGS) for primary tumors and organoids were performed. Morphologic growing pattern and immunofluorescent stains for stem cell markers (CD44 and ALDH1) and epithelial-mesenchymal marker (E-cadherin and vimentin) were investigated in immortal organoids (EC19 and EC27). Results: Both organoids passaged multiple over 1 years; 19 months (EC19) and 12 months (EC27) and are still growing. Their growing pattern is far different and maintains their growing patterns over passages. EC19 organoid is concentric and highly proliferative pattern, whereas EC27 organoid is less proliferative but highly infiltrative pattern (Figure 1). Both organoids showed partial remission after concurrent chemoradiotherapy (CCRT). Clinical stage of EC19 and EC27 was stage III and they also showed partial remission after CCRT. In clinical setting, primary lesion of EC19 was highly proliferative but not distant metastasis during followup, whereas EC27 showed distant metastasis. NGS for whole RNA showed a high similarity between primary tumor and over 5th passaged organoid (correlation: 0.9). NGS for primary tumor revealed different RNA expression between EC19 and EC27. Both organoids showed
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Su2028 NATURAL HISTORY OF PERCUTANEOUS ENTERAL GASTROSTOMY TUBE IN PATIENTS WITH AMYOTROPHIC LATERAL SCLEROSIS Mohanad T. Al-Qaisi, Richard Gerkin, Logan Cobb, Mary Chew, Katherine Petersen, Rakesh Nanda Background: Amyotrophic lateral sclerosis (ALS) is a devastating disease that puts a grave burden on patients' quality of life. It is associated with rapid weight loss due to dysphagia and increased metabolism, which eventually leads to early mortality. Percutaneous endoscopic gastrostomy (PEG) is usually performed for nutritional support for a variety of reasons. Studies that looked at effects of PEG tubes on mortality are limited. Aim: We aimed to examine the baseline characteristics, functional status and overall survival for patients with ALS. Methods: Retrospective cohort study patients diagnosed with ALS at a large VA specialty ALS clinic in a metropolitan city between 01/2010 and 08/2017 with at least three months of follow up. Patients' demographics (age, gender, baseline functional status, weight and BMI at time of diagnosis,) were gathered. Use of riluzole, weight and BMI at 1 year were ascertained using chart review on follow up visits at the ALS clinic. Survival analysis was calculated using a Cox regression model. A p-value of 0.05 was considered statistically significant. Results: 97 consecutive patients were included in the study. 52 (53%) patients received PEG tube during study time. Baseline characteristics are in table 1. Overall, our patients lived a median of 750 days from diagnosis (95% CI 465-1034). Interestingly, PEG patients had lower median survival (730 days from diagnosis (95% CI 593-867)) compared to non-PEG patients (909 days from diagnosis (95% CI 409-1409), however this did not reach statistical significance (p = 0.70). PEG patients were younger and had lower median baseline BMI (24.3 kg/m2(+/-4) compared to non-PEG patients (28.1 kg/m2(+/-5). After 1 year, PEG patients' median BMI dropped to 22.5 kg/m2(+/-5), still less than non-PEG patients (26.9 kg/m2(+/-6). Conclusion: PEG placement does not significantly affect patient's survival in patients with ALS. More studies are required to help understand the effects of PEG tube placement in patients with ALS.
Age yrs (SD) Gender Male N (%) Baseline BMI kg/m2 (SD) Baseline weight kg (SD) BMI after 1 year from diagnosis kg/ m2 (SD) Weight after 1 year from diagnosis kg (SD) Median follow up days (SD) Baseline functional status: Wheelchair N (%) Riluzole N (%) Aspiration N (%)
PEG (n=52) 65.0 (10.0) 49 (94.2)
No PEG (n=47) 69.8 (7.5) 44 (97.8)
p-Value 0.03 0.62
24.3 (4)
28.1 (5)
0.39
74.8 (13.6)
90.2 (17.0)
0.39
22.5 (5)
26.9 (6)
0.46
69.6 (19.7)
87.5 (19.3)
0.40
667 (674)
455 (460)
0.36
36 (78.3)
21 (60.0)
0.08
34 (68.0) 29 (76.3)
31 (68.9) 4 (15.4)
0.93 <0.0001
Su2039 USE OF PATIENT-DERIVED TUMOR XENOGRAFT MODEL TO GUIDE TREATMENT DECISIONS FOR PATIENTS Yanping Zhu, Chun-Mei Zhao, Yanpeng Hao, Zhiqiang Wang, Li Wang, Duan Chen Background/aim: Patient-derived tumor xenograft (PDTX) models are established by direct engraftment of patient-derived tumor fragments into immunocompromised mice. Approx. 20, 000 PDTXs have been so far created in academic and industry labs in US and Europe for basic and preclinical cancer research, such as biomarker discovery, drug screening for personalized medicine, understanding of drug-resistance mechanisms and novel therapy development. The aim of our work was to use PDTXs to guide clinical decisions for patients. Materials and Methods: From January 2015 to November 2018 at 5 clinical centers in China, we have collected tumor samples by surgery, biopsy and/or endoscopy from 723 patients with esophageal cancer (12 cases), gastric cancer (84), colorectal cancer (65), liver cancer (294), pancreatic cancer (46), sarcomas (94), gynecological cancer (63), lung cancer (26), and others (34). In 434 out of 723 PDTXs, the primary tumors (P0) grew to 800 mm3 in mice, and then re-implanted into mice to produce the first passage (P1). When the P1 tumors grew to 100–150 mm3, clinical tests of treatment regimens were conducted. It took 3-5 months from tumor sampling to clinical report. The cost was about 200 000 RMB (approx. 30,000 USD) per patient. In the remaining 289 PDTX, the P0 tumors grew in a “tumor-microenvironment matrix” that was implanted in mice, named as superPDTX (patent application no. 201810916601.5). Biomarkers of superPDTX included Ki67 index and pathological scores. It took 2 weeks at a cost of 30 000 RMB (or 5 000 USD) per patients. In addition, whole exome sequencing (WES) was performed in 8 superPDTXs. Results: The overall engraftment rate of PDTX (P1) was 56.2%, e.g. esophageal cancer was 45.5%, gastric cancer was 63.9%, colorectal cancer was 62.2%, liver cancer was 53.8%, and pancreatic cancer was 52.6%. A cohort study of 21 PDTXs/patients showed that the consistency of treatment effects was 85.7% (18/21) in which 15 out of 17 was validated in co-clinical trial and 3 out of 4 was in predictive testing. A total of 289 superPDTXs were used for pharmaceutical drug screening (61 cases) and clinical testing (228 patients, including 160 GI cancers). The successful rate of superPDTX (P0) clinical testing was 92%. Another cohort of 22 superPDTXs/patients receiving 28 treatment regimens showed a consistency rate of 82.1% (23/28) in which 8 out of 13 was validated in co-clinical trial and 15 out of 15 was in predictive testing. Up-to-date, we have banked the PDTX tissue samples (P0-P2) of > 35 000. WES data showed that >90% mutations identified in primary tumors were present in the corresponding superPDTX samples (P0). Conclusion: Based on our results and clinical practice, we suggested that superPDTX testing could be used routinely to enhance clinical decision making for personalized cancer care. Su2040 DIVERSE TUMOR SUSCEPTIBILITY IN COLLABORATIVE CROSS MICE: IDENTIFICATION OF A NEW MODEL FOR HUMAN GASTRIC TUMORIGENESIS Pin Wang, susan Celniker, Xiaoping Zou, Bo Hang, Antoine M Snijders, Jianhua Mao Objective: The Collaborative Cross (CC) is a mouse population model with diverse and reproducible genetic backgrounds used to identify novel disease models and genes that contribute to human disease. Since spontaneous tumor susceptibility in CC mice remains unexplored, we assessed tumor incidence and spectrum. Design: We monitored 293 mice from 18 CC strains for tumor development. Genetic association analysis and RNA-sequencing were used to identify susceptibility loci and candidate genes. We analyzed genomes of gastric cancer patients to evaluate the relevance of genes identified in the CC mouse model and measured the expression levels of ISG15 by immunohistochemical staining using a gastric adenocarcinoma tissue microarray. Association of gene expression with overall survival (OS) was assessed by Kaplan-Meier analysis. Results: CC mice displayed a wide range in the incidence and types of spontaneous tumors. More than 40% of CC036 mice developed gastric tumors within one year. Genetic association analysis identified Nfκb1 as a candidate susceptibility gene, while RNA-sequencing analysis of non-tumor gastric tissues from CC036 mice showed significantly higher expression of inflammatory response genes. In human gastric cancers the majority of human orthologs of the 166 mouse genes were preferentially
Su2029 USEFULLNESS OF BRANCHED-CHAIN AMINO ACID-ENRICHED ENTERAL NUTRITION IN THE ENDOSCOPIC TREATMENT OF ESOPHAGEAL VARICES Yuichi Kojima, Toshihisa Takeuchi, Shinya Nishida, Noriaki Sugawara, Kazuhiro Ota, Satoshi Harada, Kazuhide Higuchi [Background] Fasting during the endoscopic treatment of esophageal varices may deteriorate the nutritional statuses of patients with cirrhosis, suggesting the need for nutritional intervention to maintain the hepatic functional reserve. [Objective] To examine the usefulness of branched-chain amino acid-enriched enteral nutrition in the endoscopic treatment of esophageal varices. [Methods] Of patients who underwent endoscopic treatment for esophageal varices in Osaka Medical College Hospital between January 2014 and June 2017, those who did not take a branched-chain amino acid (BCAA) preparation at the beginning of the
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in 1 patient (7.7%). We recorded baseline characteristics such as age, sex, comorbidities, indications for PTEG, and preoperative biomarkers. We evaluated clinical outcomes, such as adverse events, postoperative length of stay, place of discharge, readmission rates, and mortality. Results: Of the 13 patients, 10 underwent PTEG for receiving nutrition and 3 underwent PTEG to reduce the pressure on the intestinal tract. Underlying conditions that required PTEG were gastric resection in 10 patients (76.9%), left diaphragm disorder in 2 patients (15.4%), and an interposing transverse colon or liver between the abdominal wall and anterior gastric wall in 1 patient (7.7%). PTEG was successful in all patients. The mean procedure time of PTEG was 47.9±22.3 minutes. As a postoperative adverse event, 1 patient (7.7%) dislodged the tube accidentally and required reinsertion of a new tube. No mortality was observed at 30 days. The mean postoperative length of stay was 48.5 (12–69) days. Two patients (15.4%) were discharged home, and 8 (61.5%) were discharged to long-term care hospitals. Three patients (23.1%) died of primary diseases before discharge; 1 patient (7.7%) was able to receive full nutrition orally, and this patient's percutaneous transesophageal gastric tube was dislodged on postoperative day 81. Conclusions PTEG is an effective and safe method to provide nutrition or reduce the pressure of the intestinal tract. PTEG may be an alternative technique when gastrostomy is impossible.
AGA Abstracts
altered by amplification or deletion and were significantly associated with OS. Higher expression of the CC036 inflammatory response gene signature is associated with poor OS. Finally, ISG15 protein is elevated in gastric adenocarcinomas, and correlated with shortened patient OS. Conclusions: CC strains exhibit tremendous variation in tumor susceptibility, and we present CC036 as a spontaneous laboratory mouse model for studying human gastric tumorigenesis.
Su2044 EXOSOMES RELEASED FROM PATIENT-DERIVED ORGANOIDS CONTAIN DIFFERENT MICRORNA PROFILES BETWEEN COLORECTAL CANCER AND ADENOMA Hiroshi Nagai, Masatake Kuroha, Atsushi Masamune Exosomes are membrane-enclosed nanovesicles and have been increasingly recognized as important cell communications by transmitting microRNA (miRNA). No previous studies have addressed the exosomal miRNA profile in colorectal adenoma (CRA) due to the difficulty of long-term culture of CRA. Development of a three-dimensional organoid culture system allows us to perform long-term expansion of epithelial organoids of human CRA and CRC. The aim of this study was to clarify the differences in exosomal and cellular miRNA profiles between organoids derived from CRA and CRC. We established six CRC patient-derived organoids and eight CRA patient-derived organoids. Exosomes were prepared from conditioned medium of patient-derived organoids. We analyzed the exosomal and cellular microRNA expression profiles by miRNA microarray. We found different miRNA expression profiles between CRC- and CRA-derived organoids, and that both exosomal and cellular miR-1246 expression was upregulated in CRC-derived organoids compared to CRA-derived organoids. Alteration of miR-1246 expression by miR-1246 mimic or inhibitor accordingly increased or decreased cell proliferation in HT-29 CRC cell line, respectively. In conclusion, we here reported the first study dissecting the miRNA profile in exosomes in CRA- and CRC-derived organoids. Upregulation of miR-1246 might play a role in increased cell proliferation in colorectal adenoma-carcinoma transition.
Su2042 DERIVATION OF OESOPHAGEAL ORGANOIDS TO RECAPITULATE THE HETEROGENEITY OF PRIMARY TUMOURS AND PROVIDE A MODEL SYSTEM FOR PRECISION THERAPEUTICS Xiaodun Li, Hayley Francies, Ahmad Miremadi, Amber Grantham, Nicola Grehan, Ginny Devonshire, Mathew Garnett, Rebecca C. Fitzgerald Oesophageal adenocarcinoma (EAC), the dominant subtype of oesophageal cancer in western countries, has increased six-fold in the last 30 years. Despite concerted efforts to improve patient management the overall 5-year survival is still below 15% due to late diagnosis and limited response to traditional therapy. Whole genome sequencing efforts of the ICGC have enabled us to better characterise the genomic landscape which is dominated by copy number changes and structural rearrangements with enormous heterogeneity in point mutations (SNVs) and InDels. The genomic heterogeneity has made it difficult to ascribe rational targeted therapy. However, the more homogeneous mutational signatures and integrated analysis of -omits and expression changes suggest new therapeutic hypotheses. In order to test these hypotheses model systems are required. To overcome the drawback of historical 2D lines with missing germline status and unreliable EAC mouse models, we have developed primary EAC organoids with matched WGS data for the primary tumour and germline. To maxim the power of cellular model to recapitulate this extreme heterogeneous cancer, to date 27 organoids have been developed with a lifespan over 6 months, which provides the biggest EAC cell model collection so far. These organoids are representative of the clinical disease: 68% of organoids are from male patients with an average age of 67 years; 18 of 27 patients received chemo treatment and12 of these chemo treated patients showing chemo resistance (Tumour Regression Grade score > 3). Based on our recent description of the genomic landscape of 551 EAC patients, 74 of 76 driver mutations could also be found in this organoid cohort. For example, 78% of harbour TP53mutations, 33% ARID1A, and 29% SMAD4. A thorough phenotypic and molecular characterisation has confirmed that the organoids faithfully recapitulate the parental tumour at a genomic protein level and have enabled us to study clonality, cell kinetics and tumour polarity. Organoid models in combination with genomic characterisation of the matched primary provides a functional platform to develop rational therapeutic choices in EAC.
Su2045 VASCULAR SHUT DOWN EFFECTS OF PHOTODYNAMIC THERAPY WITH TALAPORFIN Taketo Suzuki, Mamoru Tanaka, Hiroshi Ichikawa, Hirotada Nishie, Hiromi Kataoka Background: Photodynamic therapy (PDT) is an attractive modality for cancer therapy because of minimally invasive and minimally toxic. PDT consists of the intravenous administration of a photosensitizer, followed by activation with a specific wavelength light. Activation of the photosensitizer causes the conversion of molecular oxygen into various reactive oxygen species that directly induce the death of the tumor cells or shut down the tumor-associated vasculature. However, the mechanism of the vascular shut down is not clear. Here, we evaluated that PDT with Talaporfin (mono-l-aspartyl chlorin6, Laserphyrin®) shut down existing tumor vessels. Materials and Methods: (1) Talaporfin was administered to human umbilical vein endothelial cell lines (HUVEC) followed by irradiation with a 664 nm LED. The cell death-inducing effects were analyzed by WST-8 assay and half maximal inhibitory concentration (IC50) was calculated. (2) Tubulin and F-actin (actin stress fiber) were analyzed by immunofluorescent staining in HUVEC treated with PDT. The phosphorylation of myosin light chain (MLC) was examined by Western blotting. And we evaluated the relationship with Rho-GTP pathway by the Rho-kinase inhibitor; Y-27632. (3) Xenograft tumor mouse models were established with colon cancer cell lines (HCT116). After Talaporfin intravenous injection, tumors were illuminated with the diode laser system (664nm). Tumor blood flow was measured by a laser speckle blood flow analyzing system; OMEGA ZONE2. Temporal changes of the flow were compared with the positive control; Combretastatin A-4 phosphate (CA4P). Result: (1) PDT with Talaporfin induced the cell death effectively in HUVEC. (IC50 = 13.6 cˇmol/l). (2) PDT induced the depolymerization of tubulin and the polymerization of F-actin because of destruction of the microtubule. But the Rho-kinase inhibitor Y-27632 prevented the polymerization of F-actin. It indicated that PDT induced the vascular shut down effect by polymerization of F-actin via the Rho-GTP pathway. (3) PDT with Talaporfin decreased the blood flow in tumors but not in the neighboring healthy tissue. And the blood flow rate significantly decreased compared with CA4P. Conclusions: PDT with Talaporfin induced vascular shut down effect by activating of Rho-GTP pathway leading to F-actin polymerization in vitro and decreased the tumor blood flow significantly in vivo. In summary, we could show the vascular shut down of PDT with Talaporfin which suggested as an important part of its antitumor effects.
Su2043 A NOVEL CANINE INTESTINAL ORGANOID MODEL TO CHARACTERIZE DOSE-EXPOSURE-RESPONSE OF CHEMOTHERAPEUTIC DRUGS Dana C. Borcherding, Yoko M. Ambrosini, Todd Atherly, Samantha Thomson, Larry W. Wulf, David Borts, Albert Jergens, Karin Allenspach, Jonathan P. Mochel Background: Stem cell-derived 3D organoids have emerged as a cutting-edge cell culture technology for drug discovery, toxicity screening, and testing for personalized medicine. Given that gastro-intestinal (GI) toxicity is one of the commonly reported side effects of chemotherapeutics, it is important to evaluate the dose-exposure-GI toxicity response relationship of anticancer agents in a scalable in vitro system prior to their use in patients. To bridge the knowledge gap between rodents and humans, our lab has developed a three dimensional (3D) canine GI organoid (enteroid and colonoid) system for testing of chemotherapeutic drugs. Large animal models, such as the dog, are indeed more representative of humans than mice as they have a relatively large body size, longer life span, more closely resemble human GI physiology and develop spontaneous, analogous diseases including colorectal cancer (CRC). The objective of this proof-of-concept study was to demonstrate how canine organoids can be used to establish drug dose-exposure-toxicity relationships using Doxorubicin as a testbed. Methods: Canine 3D intestinal organoids cultured in matrigel were exposed to control or Doxorubicin for 2 days before matrigel was dissolved by Matrisperse. Doxorubicin concentration in organoids was then determined by liquid chromatography mass spectrometry (LC-MS) and compared to a 6-point standard curve (50 to ~14,000 pg range) to determine Doxorubicin concentration within organoid samples. The toxicity of Doxorubicin on 3D canine organoids in matrigel matrix was determined after 48 hours of incubation with Doxorubicin by MTT assay. Results: were compared to the cytotoxicity of Doxorubicin (1-1000 nM) in two CRC cell-lines, HCT-116 and HT-29, by MTT assay, with data evaluated by ANOVA and t-tests. Results: Exposure of canine organoids to Doxorubicin was confirmed by LC-MS, with uptake of about 10% or ~34 ng per 12 wells of organoids after incubation with 100 nM Doxorubicin for 2 days. We observed a mild, dose-dependent cytotoxicity of ~15-25% by 100-1000 nM Doxorubicin (p<0.05) after 2 days in small intestinal (jejunum) organoids from a healthy dog by the MTT assay. In comparison, Doxorubicin significantly (p<0.05) reduced cell viability in two CRC cell-lines (HCT-116 and HT-29) to about 40-60% and 10-20% of control for 100 and 1000 nM Doxorubicin, respectively. Conclusions: We confirmed that Doxorubicin was functional, and, as expected, exerted only a mild cytotoxic effect on the non-cancer intestinal organoids but showed much higher cytotoxicity on the fast proliferating CRC cells. These studies establish proof-of-concept that canine intestinal organoids can be used for in vitro doseexposure-cytotoxicity characterization of chemotherapeutic drugs.
AGA Abstracts
Su2046 MORPHOLOGIC DIFFERENCE OF PATIENTS-DERIVED IMMORTAL ESOPHAGEAL CANCER ORGANOIDS WAS ASSOCIATED WITH MOLECULAR AND POST-GENOMIC DIFFERENCE Su Youn Nam, Hee Jeong Lim, Su In Kim Background: Primary organoid have advantages in providing more physiologically relevant and predictive data for in-vivo response. Previously, we showed high agreement between clinical response and response in patents-derived primary organoid. We produced immortal patient derived esophageal cancer organoid and investigated the morphologic and molecular characteristics supporting the correlation between primary tumor and organoid. Method: Patient-derived primary organoid culture was performed using tumor tissues acquired from esophageal squamous cell cancer. We cultured them up to 1000um diameter and then dissected into 3-4 organoids. Next-generation sequencing (NGS) for primary tumors and organoids were performed. Morphologic growing pattern and immunofluorescent stains for stem cell markers (CD44 and ALDH1) and epithelial-mesenchymal marker (E-cadherin and vimentin) were investigated in immortal organoids (EC19 and EC27). Results: Both organoids passaged multiple over 1 years; 19 months (EC19) and 12 months (EC27) and are still growing. Their growing pattern is far different and maintains their growing patterns over passages. EC19 organoid is concentric and highly proliferative pattern, whereas EC27 organoid is less proliferative but highly infiltrative pattern (Figure 1). Both organoids showed partial remission after concurrent chemoradiotherapy (CCRT). Clinical stage of EC19 and EC27 was stage III and they also showed partial remission after CCRT. In clinical setting, primary lesion of EC19 was highly proliferative but not distant metastasis during followup, whereas EC27 showed distant metastasis. NGS for whole RNA showed a high similarity between primary tumor and over 5th passaged organoid (correlation: 0.9). NGS for primary tumor revealed different RNA expression between EC19 and EC27. Both organoids showed
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