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SummaryTo determine the maximum tolerated dose (MTD) and recommended dose (RD) of orally-administered bendamustine in Japanese patients with advanced solid tumors. The optimal dosing schedule, safety, pharmacokinetics, and preliminary antitumor effects were also evaluated. A multicenter, open-label trial with a standard 3 + 3 design and dose escalation by dose-limiting toxicity (DLT) was conducted. The treatment schedules were once daily for 7, 14, and 21 days every 3 weeks as one cycle. The total dose per cycle was increased from 175 to 840 mg/m2. Eighteen patients were enrolled in this study. DLT occurred in one of six patients at 75 mg/m2/day × 7 days, and one of three patients at 37.5 mg/m2/day × 14 days and 25 mg/m2/day × 21 days. However, the delayed recovery from a decrease in neutrophil or platelet count hampered the start of subsequent treatment cycles, and the trend was more prominent at 37.5 mg/m2/day × 14 days and 25 mg/m2/day × 21 days than in 75 mg/m2/day × 7 days. MTD was determined as 75 mg/m2/day × 7 days to allow acceptable hematologic recovery. The pharmacokinetics of orally-administered bendamustine were generally dose-dependent; however, the inter-individual variability is relatively large. The major adverse events were hematologic toxicities; gastrointestinal disorders were generally mild. Adverse drug reactions did not lead to the discontinuation of the drug. A partial response was observed in two of six patients (prostatic small cell carcinoma and thymic carcinoma) at 75 mg/m2/day × 7 days. The RD and optimal dosing schedule of orally-administered bendamustine was 75 mg/m2 once daily for 7 days every 3 weeks for the treatment of advanced solid tumors. (Trial registration number ClinicalTrials.gov NCT03604679. Registration date July 27, 2018).

C-Glycosides, in which a sugar moiety is linked via a carbon-carbon (C-C) bond to a non-sugar moiety (aglycone), are found in our food and medicine. The C-C bond is cleaved by intestinal microbes and the resulting aglycones exert various bioactivities. Although the enzymes responsible for the reactions have been identified, their catalytic mechanisms and the generality of the reactions in nature remain to be explored. Here, we present the identification and structural basis for the activation of xenobiotic C-glycosides by heterocomplex C-deglycosylation enzymes from intestinal and soil bacteria. They are found to be metal-dependent enzymes exhibiting broad substrate specificity toward C-glycosides. X-ray crystallographic and cryo-electron microscopic analyses, as well as structure-based mutagenesis, reveal the structural details of these enzymes and the detailed catalytic mechanisms of their remarkable C-C bond cleavage reactions. Furthermore, bioinformatic and biochemical analyses suggest that the C-deglycosylation enzymes are widely distributed in the gut, soil, and marine bacteria., In C-glycosides the sugar moiety is linked through a carbon-carbon bond to the non-sugar moiety, which can be cleaved by intestinal microbes. Here, the authors use bioinformatics analysis to identify C-glycoside deglycosidase enzymes in intestinal and soil bacteria, biochemically characterise them and determine their structures and probe catalytic important residues in mutagenesis experiments.

Background Implementation of personalized medicine requires the accessibility of tumor molecular profiling in order to allow prioritization of appropriate targeted therapies for individual patients. Our aim was to study the role of comprehensive genomic profiling assays that may inform treatment recommendations for patients with solid tumors. Materials and Methods We performed a prospective study to evaluate the feasibility of application of the FoundationOne CDx panel—which detects substitutions, insertions and deletions, and copy number alterations in 324 genes, select gene rearrangements, and genomic signatures including microsatellite instability and tumor mutation burden (TMB)—to patients with advanced or recurrent solid tumors before its approval in Japan. Results A total of 181 samples were processed for genomic testing between September 2018 and June 2019, with data being successfully obtained for 175 of these samples, yielding a success rate of 96.7%. The median turnaround time was 41 days (range, 21–126 days). The most common known or likely pathogenic variants were TP53 mutations (n = 113), PIK3CA mutations (n = 33), APC mutations (n = 32), and KRAS mutations (n = 29). Among the 153 patients assessed for TMB, the median TMB was 4 mutations/Mb, and tumors with a high TMB (≥10 mutations/Mb) were more prevalent for lung cancer (11/32) than for other solid tumor types (9/121, Fisher's exact test p < .01). No clear trend toward increased efficacy for immune checkpoint inhibitor (ICI) monotherapy or ICI combination chemotherapy in patients with a high programmed cell death–ligand 1 tumor proportion score or a high TMB was apparent. Among the 174 patients found to harbor known or likely pathogenic actionable alterations, 24 individuals (14%) received matched targeted therapy. Conclusion The FoundationOne CDx assay was performed with formalin‐fixed, paraffin‐embedded tumor specimens with a success rate of >95%. Such testing may inform the matching of patients with cancer with investigational or approved targeted drugs. Implications for Practice This prospective cohort study was initiated to investigate the feasibility and utility of clinical application of FoundationOne CDx. A total of 181 samples were processed for genomic testing between September 2018 and June 2019, with data being successfully obtained for 175 of these samples, yielding a success rate of 96.7%, and 24 individuals (14%) received matched targeted therapy., Comprehensive genomic profiling assays can identify multiple actionable genetic alterations that may inform treatment recommendations for patients with solid tumors. This article evaluates the feasibility of the application of the FoundationOne CDx panel to patients with advanced or recurrent solid tumors.

Significance Microbial degradation of C -glycosides is only reported from human intestinal bacteria. This study isolated a C -glycoside–catabolizing bacteria, Microbacterium sp. 5-2b, from soil and identified a C -glycoside–metabolizing enzyme. Surprisingly, while the metabolic pathway of C -glycoside in the strain was similar to that of an intestinal one, the enzyme named CarA catalyzing the initial step of metabolism was completely different from an enzyme identified from the intestinal microorganism; CarA identified as C -glycoside 3-oxidase for C-glycoside metabolism exhibited amino acid sequence similarity to pyranose oxidase family proteins. This study clarified the structure, function, and distribution of the enzyme involved in C -glycoside catabolism and provides insight into the biogeochemical circulation of C -glycosides in nature.

C-glycosides have a unique structure, in which an anomeric carbon of a sugar is directly bonded to the carbon of an aglycone skeleton. One of the natural C-glycosides, carminic acid, is utilized by the food, cosmetic, and pharmaceutical industries, for a total of more than 200 tons/y worldwide. However, a metabolic pathway of carminic acid has never been identified. In this study, we isolated the previously unknown carminic acid-catabolizing microorganism and discovered a flavoenzyme “C-glycoside 3-oxidase” named CarA that catalyzes oxidation of the sugar moiety of carminic acid. A Basic Local Alignment Search Tool (BLAST) search demonstrated that CarA homologs were distributed in soil microorganisms but not intestinal ones. In addition to CarA, two CarA homologs were cloned and heterologously expressed, and their biochemical properties were determined. Furthermore, a crystal structure of one homolog was determined. Together with the biochemical analysis, the crystal structure and a mutagenesis analysis of CarA revealed the mechanisms underlying their substrate specificity and catalytic reaction. Our study suggests that CarA and its homologs play a crucial role in the metabolism of C-glycosides in nature.

Extramammary Paget disease (EMPD) is a rare cutaneous adenocarcinoma that usually is of epidermal origin and shows glandular differentiation and that is treated by wide local excision depending on the disease extent. For widely metastatic disease, however, a standard treatment remains to be established. Similar to breast cancer, EMPD has been found to overexpress human epidermal growth factor receptor 2 (HER2) or hormone receptors (HRs). Whereas HER2-directed therapy was recently shown to be effective for HER2-positive EMPD, the potential role of endocrine therapy for HR-positive EMPD has remained unknown. We here report a case of metastatic EMPD with HR positivity that was successfully treated with the selective estrogen receptor modulator tamoxifen. This first-line treatment of systemic metastasis resulted in durable tumor regression for > 20 months without any treatment-related toxicities. This is the first report to reveal the promise of tamoxifen as a safe and effective treatment for HR-positive metastatic EMPD.

Background: Juzentaihoto (JTT) is a Kampo prescription that has been used clinically for treating skin diseases such as atopic dermatitis in Japan. We have previously studied the anti-allergic effects of JTT on 2,4,6-trinitrochlorobenzene (TNCB)-induced contact hypersensitivity (CHS) in mice and demonstrated that it significantly suppresses ear swelling in a dose-dependent manner. However, the mechanism underlying the anti-allergic actions of JTT is obscure. Methods: We investigated the mechanism underlying the anti-allergic effects of JTT using a TNCB-induced murine CHS model and adoptive cell transfer experiments. Results: We showed that the anti-allergic effects of JTT are due to inhibition of effector T-cell activation and induction and/or activation of regulatory T cells. Furthermore, ex vivo experiments confirmed the effect of JTT on the activation of effector T cells and regulatory T cells, as interferon-γ production decreased, whereas interleukin (IL)-10 production increased, in the cultured lymphocytes obtained from 5% TNCB-sensitized mice treated with anti-CD3ε and anti-CD28 monoclonal antibodies. Flow cytometry showed that the CD4+CD25+Foxp3+, CD4+CD25+Foxp3−, and CD8+CD122+ cell population increased after oral administration of JTT. Finally, the anti-allergic effect of JTT by inducing and/or activating regulatory T cells (Tregs) was confirmed to be mediated by IL-10 through in vivo neutralization experiments with anti-IL-10 monoclonal antibodies. Conclusion: We suggested that JTT exerts anti-allergic effects by regulating the activation of effector T cells and Tregs involved in murine CHS model.

HER2‐targeted therapy, especially the anti‐HER2 antibody trastuzumab, is standard for HER2‐positive breast cancer; however, its efficacy is limited in a subpopulation of patients. HER3 ligand (heregulin)‐dependent HER2‐HER3 interactions play a critical role in the evasion of apoptosis and are therefore a target for oncotherapy to treat HER2‐positive breast cancer. The anti‐HER2 antibody pertuzumab and anti‐HER3 antibody patritumab both target this heregulin–HER3‐HER2 complex in different ways. This study examined the anticancer efficacy of dual HER2 and HER3 blockade in trastuzumab‐resistant HER2‐positive breast cancer. HER2‐positive SKBR3 or BT474 cells overexpressing heregulin (SKBR3‐HRG, BT474‐HRG) were used to evaluate the efficacy of trastuzumab, pertuzumab, and patritumab in vitro by performing cell viability, immunoblotting, and clonogenic assays. The effects of these agents were then evaluated in vivo using BT474‐HRG and an intrinsic heregulin‐expressing and HER2‐positive JIMT‐1 xenograft models. SKBR3‐HRG and BT474‐HRG cells lost sensitivity to trastuzumab, which was accompanied by Akt activation. Unexpectedly, trastuzumab in combination with pertuzumab or patritumab also showed limited efficacy toward these cells. In contrast, trastuzumab/pertuzumab/patritumab triple treatment demonstrated potent anticancer efficacy, concomitant with strong repression of Akt. Finally, in heregulin‐expressing BT474‐HRG and JIMT‐1 xenograft models, the addition of pertuzumab and patritumab to trastuzumab also enhanced antitumor efficacy leading to tumor regression. The current study found that triple blockade of HER2 and HER3 using trastuzumab, pertuzumab, and patritumab could overcome resistance to trastuzumab therapy in heregulin‐expressing and HER2‐positive breast cancer, which could be exploited clinically.

The high prevalence of obesity is a global concern. However, obesity and inflammatory bowel disease have similarities in epidemiology, diet, and gut microbial dysbiosis. An omnivorous (westernized) diet replacement with a plant-based diet in inflammatory bowel disease achieved better outcomes in both the induction and quiescent phases. Educational hospitalization providing a plant-based diet was found effective in suppressing flare-up in ulcerative colitis. This case report was designed to assess whether a plant-based diet can reduce body weight in patients with obesity during educational hospitalization for a short period of two weeks. Three men with metabolic syndrome (55, 61, and 65 years old) were included in the study with body mass index ranging from 27.0 to 31.2 kg/m2 and visceral fat area between 94.7 and 128 cm2. In Japan, a body mass index of ≥25 is defined as obesity. A lacto-ovo-vegetarian diet with brown rice (1400 or 1100 kcal/day), a plant-based diet, was provided during hospitalization for around two weeks. Patients walked for about 1 h in a park daily during the hospitalization. All three patients showed weight loss: 2.9%, 6.1%, and 6.6% at discharge. A decrease in body mass index, waist circumference, and the visceral fat area was also observed. Restriction in total calories in the form of a plant-based diet and daily 1-hour walks through an educational hospitalization for two weeks effectively reduced body weight and improved obesity-related parameters. Thus, a plant-based diet could be an effective weight-loss diet.

Frail older hospital patients are susceptible to malnutrition and iatrogenic sarcopenia. This can be linked to the decreased appetite and oral intake that can arise in largely bed-bound patients who do not get up even for rehabilitation and meals. The KT index was devised as an easy-to-use evaluation tool to address oral intake issues, and it has potential utility for expediting a multidisciplinary comprehensive rehabilitation program. To our knowledge, no reports have described real-world evidence on multidisciplinary team interventions with this tool. Herein, we report the case of a frail older patient whose oral intake improved following a KT Index-based intervention.

PURPOSE Cancer treatment can alter patient appearance, leading to psychological, social, and behavioral issues. This study aimed to investigate distress and difficulties related to appearance concerns in Japanese cancer patients and to identify information and support needs among them. METHODS We conducted a questionnaire survey using the Derriford Appearance Scale 59 (DAS59) among cancer patients with a prior history of chemotherapy, molecular targeted therapy, or immunotherapy, who were recruited from the Departments of Medical Oncology and Psychosomatic Medicine, Kindai University Hospital. RESULTS Participants were 114 patients with a mean age of 62.9 years; 70.2% were female, 86.0% had metastatic or locally advanced unresectable cancer, and 78.1% had concerns about some aspect of their appearance. Mean DAS59 full-scale score was 77.7 ± 36.4. Younger and female participants were found to have higher full-scale scores in univariate analysis (P

T790M mutation–selective EGFR tyrosine kinase inhibitors (EGFR-TKI) have demonstrated clinical benefits in non–small cell lung cancer (NSCLC) patients harboring T790M mutation, which is the major cause of resistance to EGFR-TKI. However, their efficacy is limited, possibly due to the emergence of apoptosis resistance in T790M-positive NSCLC. We previously identified Src family kinases as cooncogenic drivers along with T790M and found that the Src inhibitor dasatinib combined with an irreversible or a preclinical T790M-selective EGFR-TKI enhanced antitumor activity in T790M-positive cells. In the current study, we evaluated the efficacy of dasatinib combined with the clinically relevant T790M-selective EGFR-TKI ASP8273 or osimertinib in EGFR mutation–positive NSCLC with or without T790M mutation. A cell viability assay revealed that dasatinib had synergistic effects with these TKIs in T790M-positive cells and simultaneously inhibited Src, Akt, and Erk, which remained activated upon single-agent treatment. Dasatinib also increased the rate of apoptosis in T790M-positive cells induced by T790M-selective EGFR-TKIs, as determined by the Annexin-V binding assay; this was associated with downregulation of the antiapoptotic Bcl-2 family member Bcl-xL, a finding that was confirmed in mice bearing T790M-positive xenografts. Our results suggest that Bcl-xL plays a key role in the apoptosis resistance of T790M-positive NSCLC, and that dasatinib combined with clinically relevant T790M-selective EGFR-TKIs is potentially effective in overcoming resistance to first-generation EGFR-TKIs in NSCLC patients with acquired T790M. Mol Cancer Ther; 16(11); 2563–71. ©2017 AACR.

Introduction Anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitors (TKIs) show marked therapeutic efficacy in patients with non–small cell lung cancer (NSCLC) harboring the echinoderm microtubule-associated protein-like 4– ALK fusion gene. The effect on overall survival (OS) of sequential treatment with the first- and second-generation ALK-TKIs crizotinib and alectinib, respectively, has remained unknown. We have examined the clinical outcome of such sequential treatment in a retrospective analysis of patients with ALK -rearranged NSCLC. Materials and Methods Eleven patients with ALK -rearranged NSCLC treated with crizotinib followed by alectinib were identified. The progression-free survival (PFS) and OS for these patients were determined from a retrospective review of their medical records. Results The median PFS on crizotinib or alectinib was 6.1 months (range, 1.0-15.4 months) and 15.2 months (range, 1.0-28.3 months), respectively. The median combined PFS for both crizotinib and alectinib was 18.2 months (range, 10.4-43.7 months). Crizotinib was continued beyond radiographic evidence of progressive disease in 6 of the 11 patients, with a median duration of postprogression crizotinib treatment of 9.4 months (range, 0-20.5 months). The OS period from the diagnosis of metastatic disease or the initiation of crizotinib treatment was 51.1 months (range, 20.9-69.5 months) and 48.6 months (range, 19.8-50.1 months), respectively. Conclusion Our retrospective study has revealed durable survival for alectinib treatment after crizotinib failure in patients with ALK -rearranged NSCLC.

Background We previously reported the synergistic effect of S-1 and eribulin in preclinical models. In addition, our phase I study revealed the recommended dose for the phase II study of the combination therapy in advanced breast cancer (ABC) patients pre-treated with anthracycline and taxane. Our current study reports on the efficacy and safety of the combined use of eribulin and S-1 in patients with ABC and poor prognosis. Methods Patients with breast cancer who received prior anthracycline- and/or taxane-based therapy were assigned to receive a combination therapy of eribulin (1.4 mg/m2 on days 1 and 8, every 21 days) and S-1 (65 mg/m2, on days 1 to 14, every 21 days) for advanced/metastatic disease. All patients had at least one clinicopathological factor such as being oestrogen receptor negative, Human Epidermal Growth Factor Receptor 2 (HER2) receptor negative, presence of visceral involvement, presence of three or more metastatic sites, or having a disease-free interval shorter than 2 years. The primary endpoint was the independent-reviewer assessed objective response rate (ORR). Secondary endpoints were clinical benefit rate, disease control rate, progression-free survival (PFS), and overall survival (OS). Results This study enrolled 33 patients. Confirmed ORR was 33.3% (95% CI: 17.3 to 52.8). Median PFS was 7.5 months (95% CI: 4.0 to 14.3). Median OS time was not reached during the current experimental periods. The most common grade 3/4 adverse event was neutropenia (68.8%). Conclusions The combination of eribulin and S-1 is safe and effective for treatment in patients with ABC and poor prognosis. Trial registration Current Controlled Trials UMIN000015049, date of registration: September 5th 2014.

Medical oncologists are challenged to personalize medicine with scientific evidence, drug approvals, and treatment guidelines based on sequencing of clinical samples using next generation sequencer (NGS). Knowledge-based curation systems have the potential to help address this challenge. We report here the results of examining the level of evidence regarding treatment approval and clinical trials between recommendations made by Watson for Genomics (WfG), QIAGEN Clinical Insight Interpret (QCII), and Oncomine knowledge-based reporter (OKR). The tumor samples obtained from the solid cancer patients between May to June 2018 at Kindai University Hospital. The formalin-fixed paraffin-embedded tumor samples (n = 31) were sequenced using Oncomine Comprehensive Assay v3. Variants including copy number alteration and gene fusions identified by the Ion reporter software were used commonly on three curation systems. Curation process of data were provided for 25 solid cancers using three curation systems independently. Concordance and distribution of curated evidence levels of variants were analyzed. As a result of sequencing analysis, nonsynonymous mutation (n = 58), gene fusion (n = 2) or copy number variants (n = 12) were detected in 25 cases, and subsequently subjected to knowledge-based curation systems (WfG, OKR, and QCII). The number of curated information in any systems was 51/72 variants. Concordance of evidence levels was 65.3% between WfG and OKR, 56.9% between WfG and QCII, and 66.7% between OKR and QCII. WfG provided great number of clinical trials for the variants. The annotation of resistance information was also observed. Larger differences were observed in clinical trial matching which could be due to differences in the filtering process among three curation systems. This study demonstrates knowledge-based curation systems (WfG, OKR, and QCII) could be helpful tool for solid cancer treatment decision making. Difference in non-concordant evidence levels was observed between three curation systems, especially in the information of clinical trials. This point will be improved by standardized filtering procedure and enriched database of clinical trials in Japan.

Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) are standard therapy for EGFR-mutant non-small cell lung cancer (NSCLC). Preclinically, HER3 ligand heregulin induces resistance to EGFR-TKIs, whereas the pan-human EGFR family inhibitor afatinib remains effective. Here, we examined whether soluble heregulin levels have clinical implications for EGFR-mutant NSCLC treated with EGFR-TKIs. Soluble heregulin was immunologically measured in plasma from EGFR-mutant NSCLC patients. Cutoff values were determined by 1-year PFS ROC curve. The relationship between soluble heregulin and PFS following EGFR-TKI therapy was analyzed by Cox proportional hazards model. Seventy-three patients were enrolled: 44 were treated with 1st-generation and 29 with 2nd-generation EGFR-TKIs. Soluble heregulin levels varied (range: 274–7,138 pg/mL, median: 739 pg/mL). Among patients treated with 1st-generation EGFR-TKIs, those with high heregulin (n = 20, >800 pg/mL) had a tendency for shorter PFS than those with low heregulin (n = 24, st-generation EGFR-TKIs (HR: 1.825, 95% CI: 0.865–4.318) but not against 2nd-generation EGFR-TKIs. Soluble heregulin potentially correlates with resistance to EGFR-TKIs but not 2nd-generation EGFR-TKIs in patients with EGFR-mutant NSCLC.

// Naoki Takegawa 1 , Kimio Yonesaka 1 , Kazuko Sakai 2 , Hiroto Ueda 1 , Satomi Watanabe 1 , Yoshikane Nonagase 1 , Tatsuya Okuno 1 , Masayuki Takeda 1 , Osamu Maenishi 3 , Junji Tsurutani 1 , Taroh Satoh 4 , Isamu Okamoto 5 , Kazuto Nishio 2 , Takao Tamura 1 , and Kazuhiko Nakagawa 1 1 Department of Medical Oncology, Kinki University School of Medicine, Osaka, Japan 2 Department of Genome Biology, Kinki University School of Medicine, Osaka, Japan 3 Department of Pathology, Kinki University School of Medicine, Osaka, Japan 4 Department of Frontier Science for Cancer and Chemotherapy, Osaka University Graduate School of Medicine, Osaka, Japan 5 Center for Clinical and Translational Research, Kyushu University, Kyushu, Japan Correspondence to: Kimio Yonesaka, e-mail: yonesaka@sakai.med.kindai.ac.jp Keywords: colorectal cancer, cetuximab, acquired resistance, human epidermal growth factor receptor 2, ctDNA Received: August 21, 2015 Accepted: November 21, 2015 Published: December 02, 2015 ABSTRACT Background: Patients with metastatic colorectal cancer (mCRC) harboring wild-type KRAS benefit from epidermal growth factor receptor (EGFR)-targeted therapy. However, patients who are treated with anti-EGFR antibodies will eventually develop the resistance to those agents. HER2 amplification is one of the mechanisms conferring resistance to anti-EGFR antibody therapy and could therefore be a potential therapeutic target. The aim of this study was to detect HER2 amplification in circulating tumor DNA (ctDNA) from patients with CRC and acquired resistance to anti-EGFR antibody therapy. Results: Our data showed that 22% (4/18) of patients in the cohort exhibited HER2 amplification. One of these patients was found to be positive for HER2 amplification in matched tumor specimens collected after cetuximab therapy, at which point the patient had acquired cetuximab resistance, despite being negative for HER2 amplification prior to therapy. Methods: We analyzed plasma ctDNA using digital polymerase chain reaction (PCR) from 18 patients with CRC, who had been treated with anti-EGFR antibody-based therapy (cetuximab) and subsequently acquired resistant cetuximab. HER2 gene copy number was analyzed using fluorescence in situ hybridization in tumor samples before and after acquisition of resistance to cetuximab-based therapy. Conclusion: Analysis of plasma ctDNA by digital PCR could be useful for detecting HER2 amplification in patients with CRC who were resistant to anti-EGFR antibody therapy.

Background: Activating RET gene fusions have been reported in diverse solid tumors but Case summary: A 46-year-old Japanese woman presented with FDG-avid right axillary, right neck and mediastinal lymphadenopathy on PET-CT imaging. Ultrasound identified a hypoechoic nodule in the right breast; biopsy revealed malignant cells, ER(+), PgR(-), and HER2(-). She was diagnosed with stage IV adenocarcinoma of the right breast. Genomic testing results of a lymph node specimen identified a CCDC6-RET fusion with no other reported alterations, including none in BRCA1 or BRCA2. Consistent with local standard of care, she received treatment with tamoxifen plus goserelin from March through June 2019, but these were discontinued for progression in the right breast and new lesions in the left lower lung. Re-biopsy revealed that the molecular biology changed into triple-negative at the time of progression. On 10-Jul-19 she started treatment with selpercatinib at the recommended Phase 2 dose of 160 mg twice daily in the ongoing Phase 1/2 trial of selpercatinib. She experienced rapid clinical improvement, with resolution of right breast and neck pain and erythema. Spiral CT imaging on 31-Jul-2019 and 4-Sep-2019 demonstrated a partial response by RECIST 1.1 (tumor reductions -30% and -30%), with reduction in multiple right breast masses, axillary, neck and mediastinal lymphadenopathy and left lung metastases; repeat imaging on 30-Oct-19 demonstrated a complete response. At the time of this writing, she remains in response and on treatment for >150 days, with all adverse events grades 1-2, and most recovered and none requiring dose interruption or modification. Conclusion: Selpercatinib demonstrated anti-tumor activity in a patient with RET fusion-positive breast cancer. To our knowledge, this is the first report of a breast cancer patient with a sustained response to selective, RET-targeted therapy and adds to the diversity of RET fusion-positive tumor types that may benefit from selective RET inhibition. Citation Format: Masayuki Takeda, Satomi Watanabe, S. Michael Rothenberg, Jennifer Kherani, Pearl P. French, Elizabeth Olek. Complete response to selpercatinib (LOXO-292), a highly selective RET inhibitor, in a patient with RET fusion-positive breast cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5236.

Background Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) improve the prognosis of EGFR-mutant non-small cell lung cancer (NSCLC). However, other human epidermal growth factor receptor (HER) families contribute to EGFR-TKI resistance. The HER3 ligand heregulin is aberrantly expressed in NSCLC. Previously, heregulin genomic induction in an EGFR-mutant NSCLC cell line caused EGFR-TKI resistance, except against 2nd-generation EGFR-TKIs, which uniquely blocked the pan-HER family. However, the clinical relevance of heregulin is unclear in EGFR-mutant NSCLC. Here, we aimed to explore the implication of heregulin in patients with EGFR-mutant NSCLC treated with EGFR-TKIs. Methods Soluble heregulin was immunologically measured in the plasma of patients with EGFR-mutant NSCLC. Cut-off values were determined via 1-year progression-free survival (PFS) receiver operating characteristic curve. Relationship between soluble heregulin and PFS, after EGFR-TKI therapy, was analyzed using a Cox proportional hazards model. Results Seventy-six patients were enrolled, of which 44 were treated with 1st-generation, 29 with 2nd-generation, and 3 with 3rd-generation EGFR-TKIs. Soluble heregulin levels were found to vary (range: 274–7,138 pg/mL, median: 741.5 pg/mL). Among patients treated with 1st- and 3rd-generation EGFR-TKIs, those with high heregulin (n = 22, > 800 pg/mL) had a shorter PFS than those with low heregulin (n = 25, Conclusions Results showed soluble heregulin to potentially correlate with EGFR-TKI resistance, though not so for 2nd-generation EGFR-TKIs, in patients with EGFR-mutant NSCLC. Therefore, 2nd-generation EGFR-TKIs warrant comparative clinical examination regarding their anti-cancer efficacy in heregulin-expressing NSCLC. Legal entity responsible for the study The authors. Funding Boehringer Ingelheim. Disclosure K. Yonesaka: Honoraria (self), Research grant / Funding (institution): Boehringer ingelheim. E. Iwama: Research grant / Funding (institution): Boehringer Ingelheim. H. Hayashi: Honoraria (self), Research grant / Funding (institution): Boehringer Ingelheim. S. Suzuki: Research grant / Funding (institution): Boehringer Ingelheim. R. Kato: Research grant / Funding (institution): Boehringer Ingelheim. S. Watanabe: Research grant / Funding (institution): Boehringer Ingelheim. T. Takahama: Honoraria (self), Research grant / Funding (institution): Boehringer Ingelheim. J. Tanizaki: Honoraria (self), Research grant / Funding (institution): Boehringer Ingelheim. K. Tanaka: Research grant / Funding (institution): Boehringer Ingelheim. M. Takeda: Honoraria (self), Research grant / Funding (institution): Boehringer Ingelheim. K. Sakai: Research grant / Funding (institution): Boehringer Ingelheim. K. Azuma: Honoraria (self), Research grant / Funding (institution): Boehringer Ingelheim. Y. Chiba: Research grant / Funding (institution): Boehringer Ingelheim. S. Atagi: Honoraria (self): Boehringer Ingelheim. K. Nishio: Research grant / Funding (self), Research grant / Funding (institution): Boehringer Ingelheim. I. Okamoto: Honoraria (self), Research grant / Funding (self), Research grant / Funding (institution): Boehringer Ingelheim; Honoraria (self), Research grant / Funding (self), Research grant / Funding (institution): Bristol-Myers Squibb; Honoraria (self), Research grant / Funding (self), Research grant / Funding (institution): AstraZeneca. K. Nakagawa: Honoraria (self), Research grant / Funding (self), Research grant / Funding (institution): Boehringer ingelheim; Honoraria (self), Research grant / Funding (self), Research grant / Funding (institution): AstraZeneca; Honoraria (self), Research grant / Funding (self), Research grant / Funding (institution): Bristol-Myers Squibb.

EGFR tyrosine kinase inhibitors (TKIs) are standard therapy for EGFR-mutant non-small cell lung cancer (NSCLC); however, these tumours eventually acquire chemoresistance. U3-1402 is an anti-HER3 antibody-drug conjugate with a novel topoisomerase I inhibitor, DXd. In the current study, we evaluated the anticancer efficacy of U3-1402 in EGFR-mutant NSCLC cells with acquired resistance to EGFR-TKIs. HCC827GR5 and PC9AZDR7 are EGFR-TKI-resistant clones for gefitinib and osimertinib, respectively. U3-1402 alone or in combination with the EGFR-TKI erlotinib demonstrated potent anticancer efficacy in HCC827GR5 cells using an in vitro growth inhibition assay and in vivo xenograft mouse model. U3-1402 induced apoptosis in HCC827GR5 cells accompanying phosphorylation of histone H2A.X, a marker of DNA damage, but did not block HER3/PI3K/AKT signalling. Further, we found using flow cytometry that the cell surface HER3 expression level in HCC827GR5 cells was twice that found in HCC827 cells, indicating internalization of U3-1402 was increased in resistant cells. In addition, administration of U3-1402 notably repressed growth of EGFR-TKI osimertinib-resistant PC9AZDR7 xenograft tumours, and that PC9AZDR7 cells expressed five times greater cell surface HER3 than PC9 cells. Furthermore, using immunofluorescent microscopy, HER3 was observed predominantly in the nucleus of PC9 cells, but was localized in the cytoplasm of PC9AZDR7 cells. This finding indicates that altered trafficking of the HER3-U3-1402 complex may accelerate linker payload cleavage by cytoplasmic lysosomal enzymes, resulting in DNA damage. Our results indicate that administration of U3-1402 alone or in combination with an EGFR-TKI may have potential as a novel therapy for EGFR-TKI-resistant EGFR-mutant NSCLC.

Background: Medical oncologists are challenged to personalize medicine with scientific evidence, drug approvals, and treatment guidelines based on sequencing of clinical samples using next generation sequencer (NGS). Knowledge-based curation systems have the potential to help address this challenge. We report here the results of examining the level of evidence regarding treatment approval and clinical trials between recommendations made by Watson for Genomics (WfG), QIAGEN Clinical Insight Interpret (QCII), and Oncomine knowledge-based reporter (OKR). Patients and methods: The tumor samples obtained from the solid cancer patients between May to June 2018 at Kindai University Hospital. The formalin-fixed paraffin-embedded tumor samples (n=31) were sequenced using Oncomine Comprehensive Assay v3. Variants including copy number alteration and gene fusions identified by the Ion reporter software were used commonly on three curation systems. Curation process of data were provided for 25 solid cancers using three curation systems independently. Concordance and distribution of curated evidence levels of variants were analyzed. Results: As a result of sequencing analysis, nonsynonymous mutation (n=58), gene fusion (n=2) or copy number variants (n=12) were detected in 25 cases, and subsequently subjected to knowledge-based curation systems (WfG, OKR, and QCII). The number of curated information in any systems was 51/72 variants. Concordance of evidence levels was 65.3% between WfG and OKR, 56.9% between WFG and QCII, and 66.7% between OKR and QCII. WfG provided great number of clinical trials for the variants. The annotation of resistance information was also observed. The larger difference was observed in the clinical trials information. It was due to the different filtering process among three curation systems possibly. Conclusion: This study demonstrates knowledge-based curation systems (WfG, OKR, and QCII) could be helpful tool for solid cancer treatment decision making. Difference in non-concordant evidence levels was observed between three curation systems, especially in the information of clinical trials. This point will be improved by standardized filtering procedure and enriched database of clinical trials in Japan. Citation Format: Masayuki Takeda, Kazuko Sakai, Shigeki Shimizu, Takayuki Takahama, Takeshi Yoshida, Satomi Watanabe, Tsutomu Iwasa, Kimio Yonesaka, Shinichiro Suzuki, Hidetoshi Hayashi, Hisato Kawakami, Yoshikane Nonagase, Kaoru Tanaka, Junji Tsurutani, Kazumasa Saigo, Akihiko Ito, Tetsuya Mitsudomi, Kazuhiko Nakagawa, Kazuto Nishio. A comparative study of curated contents by knowledge-based curation system in cancer clinical sequencing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1668.

9038 Background: Previous clinical trials demonstrated that alectinib (ALEC) had a longer time-to-progression than crizotinib (CRZ) in 1st-line settings. Information on long-term overall survival (OS), however, is still limited with a few studies having reported that the sequential strategy of “CRZ followed by other ALK-inhibitor” can provide extended OS. In Japan, ALEC was approved for a 1st-line setting earlier than in other countries. Methods: We reviewed the clinical data of ALK-rearranged NSCLC patients who received CRZ or ALEC between May 2012 and Dec 2016. Patients were divided into two groups according to the first-administered ALK inhibitor, the CRZ or ALEC group. In order to evaluate the efficacy of the sequential strategy of “CRZ followed by ALEC”, the combined time to treatment failure (TTF) was calculated in the CRZ group as defined by the sum of the “TTF of CRZ” plus the “TTF of ALEC” if patients were treated with ALEC followed by CRZ. In the ALEC group, the “TTF of ALEC” was calculated. The primary endpoint is the comparison between the combined TTF in the CRZ group with the TTF in the ALEC group. Results: Of 864 patients enrolled from 61 institutions, 840 patients were analyzed. Median age was 61 (range, 20-94); 56% were female; and 95% had adenocarcinoma. There were 535/305 patients in the CRZ/ALEC group. In the CRZ group, 282 patients received ALEC after CRZ failure. The combined TTF in the CRZ group was significantly longer than TTF in the ALEC group; median, 34.4 vs 27.2 months (mo); hazard ratio (HR), 0.709 [95%CI;0.559- 0.899]; P= 0.0044. However, there was no significant difference in OS between the patients who received ALEC after CRZ in the CRZ group and the patients in the ALEC group; median, 88.4 months vs. not reached; HR 1.048 [95%CI;0.758-1.451]; P= 0.7770. In the whole population, the CRZ group had a significantly shorter OS than the ALEC group; median, 53.6 mo vs not reached HR, 1.821 [95%CI;1.372-2.415]; P< 0.0001. Conclusions: The combined TTF in the CRZ group was significantly longer than TTF in the ALEC group, however, OS benefit of sequential therapy of CRZ followed by ALEC was not shown. Clinical trial information: UMIN000028605.

Metastatic spread to the brain is common in patients with non-small cell lung cancer (NSCLC), but these patients are generally excluded from prospective clinical trials. The studies, phase III study of afatinib or cisplatin plus pemetrexed in patients with metastatic lung adenocarcinoma with EGFR mutations (LUX-Lung 3) and a randomized, open-label, phase III study of BIBW 2992 versus chemotherapy as first-line treatment for patients with stage IIIB or IV adenocarcinoma of the lung harbouring an EGFR activating mutation (LUX-Lung 6) investigated first-line afatinib versus platinum-based chemotherapy in epidermal growth factor receptor gene (EGFR) mutation-positive patients with NSCLC and included patients with brain metastases; prespecified subgroup analyses are assessed in this article.For both LUX-Lung 3 and LUX-Lung 6, prespecified subgroup analyses of progression-free survival (PFS), overall survival, and objective response rate were undertaken in patients with asymptomatic brain metastases at baseline (n = 35 and n = 46, respectively). Post hoc analyses of clinical outcomes was undertaken in the combined data set (n = 81).In both studies, there was a trend toward improved PFS with afatinib versus chemotherapy in patients with brain metastases (LUX-Lung 3: 11.1 versus 5.4 months, hazard ratio [HR] = 0.54, p = 0.1378; LUX-Lung 6: 8.2 versus 4.7 months, HR = 0.47, p = 0.1060). The magnitude of PFS improvement with afatinib was similar to that observed in patients without brain metastases. In combined analysis, PFS was significantly improved with afatinib versus with chemotherapy in patients with brain metastases (8.2 versus 5.4 months; HR, 0.50; p = 0.0297). Afatinib significantly improved the objective response rate versus chemotherapy in patients with brain metastases. Safety findings were consistent with previous reports.These findings lend support to the clinical activity of afatinib in EGFR mutation-positive patients with NSCLC and asymptomatic brain metastases.

// Yoshikane Nonagase 1 , Kimio Yonesaka 1 , Hisato Kawakami 1 , Satomi Watanabe 1 , Koji Haratani 1 , Takayuki Takahama 1 , Naoki Takegawa 1 , Hiroto Ueda 1 , Junko Tanizaki 1 , Hidetoshi Hayashi 1 , Takeshi Yoshida 1 , Masayuki Takeda 1 , Yasutaka Chiba 2 , Takao Tamura 1 , Kazuhiko Nakagawa 1 , Junji Tsurutani 1 1 Department of Medical Oncology, Kindai University Faculty of Medicine, Osakasayama, Osaka, Japan 2 Clinical Research Center, Kindai University Faculty of Medicine, Osakasayama, Osaka, Japan Correspondence to: Junji Tsurutani, email: tsurutani_j@dotd.med.kindai.ac.jp Keywords: HER2, T-DM1, resistance, heregulin, breast cancer Received: March 28, 2016 Accepted: October 01, 2016 Published: October 19, 2016 ABSTRACT Background: Overexpression of heregulin, a HER3 ligand, is one mechanism that confers resistance to the anti-HER2 agents trastuzumab and lapatinib. We investigated the impact of heregulin expression on the efficacy of HER2-targeted therapeutic agents, including trastuzumab, trastuzumab emtansine (T-DM1) and lapatinib, in vitro and in vivo and evaluated the heregulin messenger RNA (mRNA) levels in specimens from patients with HER2-positive breast or gastric cancer. Results: Cell proliferation and apoptosis assays demonstrated that heregulin conferred robust resistance to lapatinib and trastuzumab via HER3-Akt pathway activation followed by survivin overexpression; however, heregulin conferred minimal or no resistance to T-DM1 and paclitaxel. The heregulin mRNA level of one of 10 patients was up-regulated after the acquisition of resistance to trastuzumab-based therapy. Materials and Methods: SK-BR-3, NCI-N87, BT-474, MDA-MB-453, HCC1954, SNU-216 and 4-1ST cells were pharmacologically treated with recombinant heregulin or transfected with the heregulin gene. We also assessed the expression of heregulin mRNA in HER2-positive breast or gastric cancer samples before and after trastuzumab-based therapy using a RT-PCR-based method. Conclusions: mRNA up-regulation of heregulin was observed in clinical breast cancer specimens during trastuzumab-based treatment, but heregulin overexpression had a limited effect on the sensitivity to T-DM1 in vitro and in vivo.

Extramammary Paget's disease is a malignant intraepithelial carcinoma, which constitutes less than 1 % of all vulvar malignancies. Surgical resection is the first treatment of choice and standard chemotherapy has not been established for advanced or recurrent disease. Experimental and clinical studies have identified human epidermal growth receptor 2 as a potential therapeutic target. A 63-year-old male was referred for recurrent extramammary Paget's disease after surgery. Human epidermal growth receptor 2 was shown to be overexpressed and amplified by immunohistochemical analysis and fluorescence in situ hybridization analysis, respectively. After two cycles of trastuzumab monotherapy, all lymph node metastases decreased in size. However, he experienced recurrence in the lymph nodes during the seven courses of trastuzumab. As a subsequent treatment, trastuzumab was administered in combination with docetaxel and pertuzumab; clinical response was sustained for 12 months without significant adverse events.

From November 2001 to January 2002, wintering birds were censused in 30 urban woodlands in Chiba City, near Tokyo, Japan, to determine the factors other than habitat area, influencing avian species composition and diversity in urban woodlands. A group of species such as Sturnus cineraceus and Columba livia were more frequently observed in sparser woodlands. These species are not typically categorized as woodland species, and they preferred mixed habitat of tree-covered vegetation and open areas. The birds of another group, including Streptopelia orientalis and Lanius bucephalus, which tend to use tree crowns as foraging habitat were more frequently observed in the woodlands surrounded by agricultural areas rather than in those surrounded by urbanized areas. This could be because those species can move across agricultural areas, but not urban areas, so that the effect of woodland isolation is not very remarkable for them when agricultural areas surrounded woodlands. On the other hand, some species preferred open area to vegetated area. A mixture of open space and woodlands would result in woodlands of reduced value for these woodland birds. Site planning that reduces the degree of interspersion should be adopted, if woodland habitat is to be conserved or restored.

A public open space is an urban facility which has significant influence on a community's formation. This study has two purposes. 1. To clarify the significance of community formation via resident's management of public open spaces. 2. To clarify the support of local government and activity of local residents for the improvement of a community. The study area is greenway Imaigawa Ikoi-no Mizube in Yokohama city. This study consists of the questionnaire investigation of the members of the Imaigawa Ikoi-no Mizube care organization which is a voluntary organization of local residents, the questionnaire investigation of the other residents of Imai-cho, and the fieldwork at the greenway. The results of these investigations were viewed according to five factors of community formation and two indexes of community value. Through these examinations, we came to two conclusions. 1. Management by the local residents contributes to a community's formation by synthesizing the image of future, deepening the attachment to the area, and deepening and widening communication. 2. For the improvement of a community, local government support and public relations is essential to not only lead the residents to autonomous activity, but also to join the care organization.

We got many photographs of private garden at the suburb residence area by door-to-door survey, sampled at random using detailed residence maps, in Yokohama Aoba in Japan, Reading and Edinburgh in UK, Torino in Italy and Freising in Germany. We selected 100 residences and choose four photographs in each residence to represent the whole garden atmosphere. Making 22 Japanese agricultural faculty students of the University of Tokyo to be an examinee, the experiment to discriminate country of garden landscape was carried out looking at four sets of photographs of 100 private gardens. This experiment carried out two times placed the interval in six months, to the identical examinee. In the second experiments, photographs of 39 residences were altered by image processing to eliminate objects such as utility poles, chairs and so on, that were thought to give the effect to discriminate country. Assessment of private garden landscape by SD methods of ten items was also carried out one time. It was shown that Japanese students could rightly detect private gardens in Japan, though they could not distinguish gardens in UK, Italy and Germany. Some objects gave the effect to distinguish country in private gardens in Japan. There existed some images to those countries, such that the (familiar) garden with globose pruning was Japanese, the (unfamiliar) garden with lawn was Foreign or England, the (natural) garden with high trees was Germany, the (artificial) garden with paving was Italy.

The farmlands located in urban fringe areas have been segmented and abandoned under high urbanization pressure. However, there was little knowledge about quantitative tendency of farmland, which located in outer suburbs where large-scale farmland complex were dominated. By clarifying the feature of the distribution of abandoned farmlands and its occurring process in large-scale farmland complex, it was revealed that although the quantitative aspect of farmland was maintained, the quality of agricultural management was declined in these areas.

In order to find the idea and impression about the urban water related area to utilize for urban landscape designing, we conducted questionnaire surveys of junior high school students and their parents who live in urban area (Bunkyo-ku, Tokyo), asking their consciousness of water related area around them and way of using of them (frequency/activity) and analyzed by a statistic method. As a result, the following three facts have been found. 1) Students have relatively less dissatisfaction compared to their parents, which is partly related to their lower interest. 2) Parents have strong dissatisfaction in terms of the access and use of water related landscape. 3) The chances and activities of students and parents differ to each other; parents positively enjoy water landscape more than students and make the most of opportunities. These indicate that the way of facilitating the water related area needs to be the high accessibility, better usability, and with better design for parents, which also enhance and attract students’ attention.

The biological decomposition and chemical changes of leaf litter of Ginkgo biloba both at the surface and bottom layers were studied for 1 year. The changes in components such as the yields of 80% ethanol and water extractives, neutral sugars, uronic acids, Klason residues, acid-soluble phenolics, ash, nitrogen, and carbon were investigated. There were great drops of the yields of 80% ethanol extractives and water extractives in both surface and bottom mulch layers. Klason residues and sugars were the major components of the solid residues. The Klason residues lost 2.2% and 30.5% of their initial mass at the surface and bottom layers, respectively, during mulching. The total losses of the neutral sugars were 71.6% at the surface layer and 89.3% at the bottom layer. These results implied that the Klason residues were the most important inner component restraining the decomposition of the leaf litter of G. biloba and all analyzed cell wall components at the bottom layer had faster decomposition rates than those at the surface layer. Nitrogen content increased during one year of mulching, while changes in carbon contents were less than 1% throughout experiment.

道路種 (国道, 都道, 区道, 市町村道) の違いに着目し, 街路緑化政策に資する基礎的知見を得ることを目的に, 樹種別の整備の動向についてみた。東京都をスタディ・エリアとして, 樹種別の高木本数のデータを多次元尺度構成法により解析した結果, 分掌管理の始まった1970年代以降, 1) 花木や常緑樹が多用され, 規定木以外の樹種も多く用いられるようになっていること, また, 2) 道路種によって街路樹整備の進展度合いが異なり, 分掌管理によって道路種ごとに樹種の選択に特徴のみられることが明らかになった。