scholarly journals Risk of Febrile Neutropenia in Very Elderly Patients Aged ≥80 Years Who Received R-CHOP Regimen: A Nationwide Analysis in Japan

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 2530-2530
Author(s):  
Kensuke Matsuda ◽  
Taisuke Jo ◽  
Arika Nukina Shimura ◽  
Akira Honda ◽  
Yosuke Masamoto ◽  
...  
Keyword(s):  
Elderly Patients ◽  
Pharmaceutical Company ◽  
Odds Ratio ◽  
Research Funding ◽  
Antimicrobial Prophylaxis ◽  
Primary Prophylaxis ◽  
Very Elderly ◽  
Chop Regimen ◽  
Chugai Pharmaceutical ◽  
Otsuka Pharmaceutical

Abstract Background: Despite the aging society, few studies have evaluated risk factors for febrile neutropenia (FN) in the very elderly. This may be due to the difficulty of conducting a prospective study in the presence of comorbidities and a limited number of patients. We retrospectively analyzed risk factors for FN in the first cycle of rituximab-cyclophosphamide-doxorubicin-vincristine-prednisolone (R-CHOP) regimen in the very elderly aged ≥ 80 years using a nationwide inpatient database in Japan. Study Design and Methods: This study was a retrospective cohort study using the Diagnosis Procedure Combination database, a nationwide inpatient database in Japan. The database includes discharge abstracts and administrative claims data from >1200 acute-care hospitals and covers around 90% of all tertiary-care emergency hospitals in Japan. We identified patients aged ≥ 80 years old with newly diagnosed diffuse large B-cell lymphoma (DLBCL) who received the first cycle of R-CHOP regimen between July 2007 and March 2017 from the database. The initial dose intensity (IDI, %) was calculated as the average ratio of the actual dose to the 100% dose of cyclophosphamide and doxorubicin. IDI less than 80% was defined as reduced IDI. Multivariable logistic regression analysis fitted with a generalized estimating equation accounting for within-hospital clustering was performed to identify factors associated with the occurrence of FN. P<0.05 was considered statistically significant. Results: We identified 1,819 patients aged ≥ 80 years old with DLBCL who were newly diagnosed and treated with R-CHOP. The median age was 83 years (interquartile range: 82-85). A total of 73 (4%) patients received antimicrobial prophylaxis, and 270 (15%) received primary prophylaxis with G-CSF. Reduced IDI of R-CHOP regimen was performed in 1,444 (79%) patients, including 697 patients with 60-80% of IDI and 747 patients with 40-60% of IDI. FN occurred in 115 of the 1,819 patients (6.3%) with a median of 10 days (interquartile range: 8-12) after the administration of cyclophosphamide. Antimicrobial prophylaxis, primary prophylaxis with daily G-CSF, and pegfilgrastim were not significantly associated with a lower occurrence of FN in the very elderly patients (odds ratio 0.69 [95% confidence interval 0.30-2.21], 1.39 [0.73-2.64], 1.43 [0.73-2.81], respectively). Reduced IDI was significantly associated with a lower occurrence of FN (40-60%: odds ratio 0.54 [0.32-0.92]). Advanced-stage lymphoma (>stage 2) and placement of central venous catheter was significantly associated with the occurrence of FN (odds ratio 1.59 [1.02-2.49], 3.19 [1.81-5.62], respectively). Conclusion: The present nationwide study showed that IDI was reduced in most of the very elderly patients with DLBCL in a real-world clinical practice. The proportion of the very elderly patients who developed FN tended to be lower than those in previous studies with younger patients, which may be explained by reduced IDI in the very elderly. Whereas, prevention strategies with G-CSF for FN may be less effective in the very elderly. Disclosures Matsuda: Ono Pharmaceutical: Other: Lecture fee; Kyowa Kirin: Other: Lecture fee. Jo: Tsumura: Other: Lecture fee, Research Funding; AstraZeneca: Other: Lecture fee; Sanofi: Other: Lecture fee; Boehringer Ingelheim: Other: Lecture fee. Shimura: Eisai: Other: Lecture fee. Honda: Otsuka Pharmaceutical: Other: Lecture fee; Ono Pharmaceutical: Other: Lecture fee; Nippon Shinyaku: Other: Lecture fee; Jansen Pharmaceutical: Other: Lecture fee; Chugai Pharmaceutical: Other: Lecture fee; Takeda Pharmaceutical: Other: Lecture fee. Masamoto: MSD K.K.: Speakers Bureau; Eisai Co., Ltd.: Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Speakers Bureau; Takeda Pharmaceutical Company Limited.: Speakers Bureau; Chugai Pharmaceutical Company: Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Speakers Bureau; Nippon Shinyaku Co., Ltd.: Speakers Bureau; AbbVie GK: Speakers Bureau; Janssen Pharmaceutical K.K.: Speakers Bureau; SymBio Pharmaceuticals: Speakers Bureau; Bristol Myers Squibb: Speakers Bureau. Yasunaga: Pfizer: Consultancy, Other: Lecture fee; Novartis: Consultancy, Other: Lecture fee; Boehringer Ingelheim: Other: Lecture fee; Chugai Pharmaceutical: Other: Lecture fee; Tsumura: Other: Lecture fee. Kurokawa: MSD K.K.: Research Funding, Speakers Bureau; Eisai Co., Ltd.: Research Funding, Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Research Funding, Speakers Bureau; Takeda Pharmaceutical Company Limited.: Research Funding, Speakers Bureau; Chugai Pharmaceutical Company: Research Funding, Speakers Bureau; Daiichi Sankyo Company.: Research Funding, Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Research Funding, Speakers Bureau; AbbVie GK: Research Funding, Speakers Bureau; Sumitomo Dainippon Pharma Co., Ltd.: Research Funding, Speakers Bureau; Nippon Shinyaku Co., Ltd.: Research Funding, Speakers Bureau; Astellas Pharma Inc.: Research Funding, Speakers Bureau; Teijin Limited: Research Funding, Speakers Bureau; Pfizer Japan Inc.: Research Funding, Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Research Funding, Speakers Bureau.

scholarly journals Optimal Cycles of Bendamustine-Plus Rituximab Therapy for Indolent B Cell Lymphoma

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 5242-5242
Author(s):  
Kazuhiro Toyama ◽  
Toshiaki Takezaki ◽  
Akira Honda ◽  
Yasunori Kogure ◽  
Akira Chiba ◽  
...  
Keyword(s):  
B Cell ◽  
Pharmaceutical Company ◽  
Research Funding ◽  
B Cell Lymphoma ◽  
Study Cohort ◽  
Indolent Lymphoma ◽  
Chugai Pharmaceutical ◽  
Company Limited ◽  
Significant Difference ◽  
Otsuka Pharmaceutical

Backgrounds Although bendamustine-plus rituximab therapy (BR) is considered as one of the standard therapy for several indolent B cell lymphomas, the optimal cycles of BR is not still uncovered. To elucidate the optimal cycles of BR, we performed a single center retrospective study of patients with indolent lymphoma treated with BR. Methods All patients with follicular lymphoma (n=40), lymphoplasmacytic lymphoma (n=11) and mucosa associated lymphoid tissue lymphoma (n=6) who underwent BR in our institute in the period between April 2011 and September 2017 were included in this study. The clinical information including the number of repeated cycles, overall survival (OS), progression free survival (PFS), laboratory findings, backgrounds, and the response to BR were analyzed retrospectively. Rituximab was administered at day 1 and bendamustine was administered at day 1 and 2, or 2 and 3, in each 28 days cycle. In the study cohort, the number of repeated cycles was allowed up to 6. The cessation of BR was allowed after 4 cycles in the patients with response to BR, according to the discretion of attending physicians. The dosage of bendamustine was reduced to 67% and 50% in 70 to 79 years and not less than 80 years, respectively. All patients in this study were prescribed trimethoprim-sulfamethoxazole combination or pentamidine for the prevention of pneumocystis pneumonia, and acyclovir for the prevention of herpes zoster. Results In total 57 patients, the median age was 65 years (range, 37 to 83). Thirty four were male, and 23 were female. Three patients were newly diagnosed and 54 were relapsed or refractory patients. The median observation period was 51.7 months (5.1 to 83.6). The overall response rate was 86.0% (CR 54.4% and PR 31.6%). The median number of repeated cycles of BR was 4 (1 to 6). There was no significant correlation between patient characteristics and the number of repeated cycles of BR. All patients were stratified by their number of repeated cycles of BR. The early cessation group (n=17) was identified that the number was from 1 to 3, and the late cessation group (n=40) was identified that the number was from 4 to 6. The 5-year OS rates in early and late cessation groups were 56.1% and 87.0%, respectively. The 5-year PFS rates in early and late cessation groups were 31.4% and 50.6%, respectively. Both 5-year OS and PFS rates in late cessation group were significantly longer than that in early cessation group (p=0.011 and p<0.01, respectively). In late cessation group, the number of the patient who underwent 4 cycles of BR (4 cycles group) was 21, and the number of the patient who underwent 5 or 6 cycles of BR (over 4 cycles group) was 19. The 5-year OS rates in 4 cycles group and over 4 cycles group was 85.7% and 85.2%, respectively. There was no significant difference between these groups in the 5-year OS rates (p=0.58). The 5-year PFS rates in 4 cycles group and over 4 cycles group was 71.8% and 31.0%, respectively. The 5-year PFS rates in 4 cycles group were significantly longer than that in over 4 cycles group (p<0.01). The most common reason of the cessation of BR was adverse event (n=15). BR were stopped in 9 patients who achieved response after 4 or 5 cycles, and in 8 patients who became relapsed or refractory. Conclusions Our study indicated that the outcome of the patients with indolent lymphoma who stopped BR after 4 cycles was not inferior to that of the patients who stopped BR after 5 or 6 cycles. The results suggest that the cessation of BR after 4 cycles may be permissible in the patients with response to BR. Disclosures Toyama: Celgene K.K.: Speakers Bureau; Chugai Pharmaceutical Company: Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Speakers Bureau; Eisai Co., Ltd.: Speakers Bureau; Nippon Shinyaku Co., Ltd.: Speakers Bureau; Bristol-Myers Squibb: Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Speakers Bureau; Daiichi Sankyo Conpany: Speakers Bureau; Takeda Pharmaceutical Company Limited.: Speakers Bureau. Nakamura:Astellas Pharma Inc.: Speakers Bureau. Kurokawa:Shionogi & Co., Ltd: Consultancy, Honoraria; Kyowa Hakko Kirin Co., Ltd.: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Research Funding, Speakers Bureau; Chugai Pharmaceutical Company: Consultancy, Research Funding, Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Speakers Bureau; Sumitomo Dainippon Pharma Co.,Ltd.: Research Funding, Speakers Bureau; Novartis Pharma K.K.: Research Funding; Eisai Co., Ltd.: Research Funding, Speakers Bureau; Boehringer Ingelheim: Speakers Bureau; Janssen Pharmaceutical K.K.: Speakers Bureau; Yakult Honsha Company: Speakers Bureau; Pfizer Japan Inc.: Research Funding; Teijin Limited: Research Funding; Nippon Shinyaku Co., Ltd.: Research Funding; Shire Japan K.K.: Speakers Bureau; Bristol-Myers Squibb: Speakers Bureau; Takeda Pharmaceutical Company Limited.: Research Funding, Speakers Bureau; Daiichi Sankyo Conpany: Speakers Bureau; Celgene K.K.: Consultancy, Speakers Bureau; MSD K.K.: Consultancy, Research Funding, Speakers Bureau; Astellas Pharma Inc.: Research Funding, Speakers Bureau; Bioverativ Japan ltd.: Consultancy.

scholarly journals Prognostic Factors of Idiopathic Hypereosinophilic Syndrome: A Nationwide Survey in Japan

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 4192-4192
Author(s):  
Akira Honda ◽  
Kazuhiro Toyama ◽  
Yu Oyama ◽  
Kensuke Matsuda ◽  
Hideaki Mizuno ◽  
...  
Keyword(s):  
Pharmaceutical Company ◽  
Clinical Features ◽  
Research Funding ◽  
Hypereosinophilic Syndrome ◽  
Clinical Information ◽  
Nationwide Survey ◽  
Line Treatment ◽  
Chugai Pharmaceutical ◽  
Company Limited ◽  
Otsuka Pharmaceutical

Abstract Background Idiopathic hypereosinophilic syndrome (iHES) is a rare disease characterized by prolonged hypereosinophilia and organ damage without any known cause of eosinophilia. Since iHES was first proposed, the concept of the disease has continued to change, and its clinical features, optimal treatment, and prognosis have not yet been elucidated. Therefore, to clarify the clinical features and prognostic factors of iHES, we conducted a nationwide survey and collected detailed clinical information of iHES in Japan. Methods We conducted a nationwide postal questionnaire survey of iHES. A first simple questionnaire was sent to departments of hematology across the country to determine presence or absence of iHES in their department. Subsequently, a detailed questionnaire was sent to the department that responded that they had experience with iHES. After collecting the questionnaires, the validity of the diagnosis was determined. Only those that corresponded to iHES were used for subsequent analysis. In this research, iHES was defined as follows: absolute eosinophil counts more than 1,500/µL, presence of organ damage due to eosinophilia, with no known cause of eosinophilia. Allergy, collagen diseases, infection, asthma, drugs, vasculitis, and other diseases that can cause eosinophilia were ruled out. The absence of hematopoietic malignancies was confirmed by bone marrow examination, fluorescence in situ hybridization for FIP1L1-PDGFRA fusion gene, and chromosomal analysis by G-banding. Results The 1st questionnaire was sent to 492 departments of hematology, and we identified 152 patients with iHES in Japan. Among those patients, a detailed clinical information was collected from 68 patients. Of the 68 patients, 23 did not meet the criteria for iHES, and the remaining were subjected to subsequent analysis. Of the 45 patients with iHES, 27 (60%) were male, and 18 (40%) were female. The median age of diagnosis was 54 (range: 16-95) years, and the median number of involved organs per patient was 2 (range: 1-7). Symptoms caused by hypereosinophilia were consisted of systemic symptom (22, 49%), hematopoietic disorder (17, 38%), skin (16, 36%), digestive (15, 33%), respiratory (14, 31%), cardiovascular (14, 31%), and kidney (4, 9%). The median white blood cell and absolute eosinophil count were 18,800/µL (5,300-73,000/µL) and 9587/µL (2,067-63,370/µL), respectively. The median hemoglobin level was 13.3 g/dL (6.6-16 g/dL), and the median platelet count was 255 × 10 9/L (4.7-54 × 10 9/L). The median levels of lactate dehydrogenase and C-reactive protein were 299 U/L (123-972 U/L) and 0.96 mg/dL (0.02-13.9 mg/dL), respectively. Of the 45 cases, 37 (82%) required treatment, and 35 (78%) received corticosteroid as 1st line treatment. Although 28 (80%) patients responded to corticosteroid, 12 (34%) patients required subsequent 2nd line treatment, and 2 (6%) patients died. In addition, 6 patients required 3rd line treatment. Six of 45 patients died from any cause during the follow-up, and the median follow-up period for censored cases was 3.1 years (0.2-23 years). The median survival from diagnosis for all cases was 2.5 years (0.1- 23 years). In univariate analysis, hemoglobin less than 10 g/dL (P=0.02), the presence of renal symptoms (P&lt;0.001), and the presence of respiratory symptoms (P&lt;0.01) were statistically significant factors for overall survival. In multivariate analysis, hemoglobin less than 10 g/dL was a statistically significant factor for overall survival (HR, 17.2; 95% CI, 1.51-197; P=0.02). Conclusion In this nationwide survey, we clarified the clinical characteristics of iHES in Japan. In addition to clinical features at the time of diagnosis, the response rate to corticosteroid and long-term prognosis were also clarified. Furthermore, the presence of anemia was found to be a poor prognostic factor for iHES. Further accumulation of cases is necessary to establish the optimal treatment strategy for iHES. Disclosures Honda: Takeda Pharmaceutical: Other: Lecture fee; Nippon Shinyaku: Other: Lecture fee; Ono Pharmaceutical: Other: Lecture fee; Otsuka Pharmaceutical: Other: Lecture fee; Chugai Pharmaceutical: Other: Lecture fee; Jansen Pharmaceutical: Other: Lecture fee. Toyama: Celgene Corporation: Other: Lecture fee; Otsuka Pharmaceutical Co., Ltd.: Other: Lecture fee; NIHON PHARMACEUTICAL CO., LTD.: Other: Lecture fee; ONO PHARMACEUTICAL CO., LTD.: Other: Lecture fee; DAIICHI SANKYO COMPANY, LIMITED: Other: Lecture fee; CHUGAI PHARMACEUTICAL CO., LTD.: Other: Lecture fee; Takeda Pharmaceutical Company Limited: Other: Lecture fee. Matsuda: Ono Pharmaceutical: Other: Lecture fee; Kyowa Kirin: Other: Lecture fee. Komatsu: Fujifilm Wako Pure Chemical Corporation: Research Funding; Fuso Pharmaceutical Industries, Ltd.: Research Funding; Japan Tobacco Inc.: Consultancy; Otsuka Pharmaceutical Co. Ltd: Membership on an entity's Board of Directors or advisory committees, Research Funding; Novartis Pharma KK: Consultancy, Research Funding, Speakers Bureau; Shire Japan KK: Consultancy, Research Funding, Speakers Bureau; PharmaEssentia Japan KK: Consultancy, Current Employment, Research Funding; Chugai Pharmaceutical Co., Ltd.: Research Funding; Kyowa Hakko Kirin Co., Ltd.: Research Funding. Kurokawa: Nippon Shinyaku Co., Ltd.: Research Funding, Speakers Bureau; Sumitomo Dainippon Pharma Co., Ltd.: Research Funding, Speakers Bureau; Chugai Pharmaceutical Company: Research Funding, Speakers Bureau; MSD K.K.: Research Funding, Speakers Bureau; Eisai Co., Ltd.: Research Funding, Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Research Funding, Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Research Funding, Speakers Bureau; Takeda Pharmaceutical Company Limited.: Research Funding, Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Research Funding, Speakers Bureau; Astellas Pharma Inc.: Research Funding, Speakers Bureau; Daiichi Sankyo Company.: Research Funding, Speakers Bureau; Teijin Limited: Research Funding, Speakers Bureau; Pfizer Japan Inc.: Research Funding, Speakers Bureau; AbbVie GK: Research Funding, Speakers Bureau.

scholarly journals Novel Recurrent Mutations in Eldheim-Chester Disease Patients Identified By Whole Exome Sequencing and Whole Genome Sequencing

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 3123-3123
Author(s):  
Yu Oyama ◽  
Akira Honda ◽  
Kensuke Matsuda ◽  
Hideaki Mizuno ◽  
Kazuki Taoka ◽  
...  
Keyword(s):  
Pharmaceutical Company ◽  
Research Funding ◽  
Braf V600e Mutation ◽  
Braf V600e ◽  
Driver Mutations ◽  
Whole Genome ◽  
Chugai Pharmaceutical ◽  
Whole Exome ◽  
Otsuka Pharmaceutical ◽  
Chester Disease

Abstract Background: Erdheim-Chester disease (ECD) is a type of systemic, non-Langerhans histiocytic disorder characterized by diffuse organ damage with infiltration of CD68-positive, CD163-positive, and CD1a-negative histiocytes. Although most patients have mutations associated with the hyperactivation of the MAPK pathway, including BRAF, MAP2K1, and NRAS, the remaining have no known driver mutations. So far, there is no specific target of treatment for them. To elucidate novel driver mutations and establish new treatment strategies in these patients, we performed whole-exome sequencing (WES) and whole-genome sequencing (WGS) using patient samples with ECD. In addition, we performed WES with multi-organ lesions in a patient with ECD to reveal the mutational profiles of each organ lesion. Methods: We performed a nationwide survey and collected clinical samples of ECD in Japan. We collected 22 samples of ECD lesions from 15 adult patients. All cases were pathologically proved. Twenty of 22 samples were formalin-fixed and paraffin-embedded tissue (FFPE) and were examined by WES. For WGS, DNA was extracted from 2 raw samples of ECD lesions. In addition, we collected multi-organ lesions in 3 patients. One patient developed myelodysplastic syndrome (MDS) and subsequent acute myeloid leukemia (AML). Therefore, we also performed WES with each bone marrow (BM) sample to reveal the relationship between ECD and these myeloid malignancies. We used 6 samples of peripheral blood, 1 BM sample, 1 skin sample, and 1 oral mucosa sample as normal controls. Result: We detected known driver mutations in seven of 15 cases (47%). Among them, BRAF V600E was detected in 5 cases (8 samples), MAP2K1 C121S in 1 case (1 sample), and NRAS Q61R in 1 case (2 samples) by WES and WGS. A mean of 69 nonsynonymous mutations per patient was identified in normal-tumor analysis (range, 2-491) and 188 in tumor-only analysis (range, 17-3598) of WES, and 3134 in normal-tumor analysis (range, 2588-3680) of WGS. The median variant allele frequency (VAF) for the 11 samples with known activating kinase mutations identified by WES and WGS was 14.4% (range, 6.3-34.7). We could not detect known driver mutations in the other 8 cases (53%). Therefore, to reveal novel driver mutations, we focused on these 8 cases. Notably, EPHA2 P786L, MYBPC3 D798N, TDRD5 P115L, and TCEAL4 F17L mutations are recurrently found in 2 out of the 8 cases, suggesting new driver mutations are contained in these. The VAFs of EPHA2 P786L were 9.1% and 9.3%, MYBPC3 D798N 5.5% and 11.9%, TDRD5 P115L 10.2% and 18.7%, and TCEAL4 F17L 7.5% and 9.0% in each case. We also analyzed multiple organ samples of a case with BRAF V600E mutation (5 samples of ECD lesions, BM with MDS, BM with AML, and skin as normal control). Interestingly, BRAF V600E mutation was identified in 3 samples (bone lesion, heart, and intestine) but was not in other samples (kidney lesion, dura matter, BM tumor, BM with MDS, and BM with AML), suggesting mutational profiles are different depending on the organ of the lesion. Conclusion: Our nation-wide analysis revealed that no well-known driver mutations were found in more than half of the ECD cases. We identified EPHA2 P786L, MYBPC3 D798N, TDRD5 P115L, and TCEAL4 F17L mutations as candidates of novel driver mutations in ECD. To elucidate the role of these mutations in pathogenesis of ECD, further functional analyses are warranted. In addition, we revealed heterogeneity of mutational profile of multi lesions in an ECD patient with BRAF V600E mutation. It is noteworthy that mutational profiles might be different depending on the organ of the lesion. Disclosures Honda: Chugai Pharmaceutical: Other: Lecture fee; Ono Pharmaceutical: Other: Lecture fee; Nippon Shinyaku: Other: Lecture fee; Takeda Pharmaceutical: Other: Lecture fee; Otsuka Pharmaceutical: Other: Lecture fee; Jansen Pharmaceutical: Other: Lecture fee. Matsuda: Kyowa Kirin: Other: Lecture fee; Ono Pharmaceutical: Other: Lecture fee. Taoka: ONO PHARMACEUTICAL CO., LTD.: Other; Chugai Pharmaceutical Company: Other. Kurokawa: Teijin Limited: Research Funding, Speakers Bureau; Chugai Pharmaceutical Company: Research Funding, Speakers Bureau; Takeda Pharmaceutical Company Limited.: Research Funding, Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Research Funding, Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Research Funding, Speakers Bureau; Eisai Co., Ltd.: Research Funding, Speakers Bureau; Astellas Pharma Inc.: Research Funding, Speakers Bureau; MSD K.K.: Research Funding, Speakers Bureau; Nippon Shinyaku Co., Ltd.: Research Funding, Speakers Bureau; Daiichi Sankyo Company.: Research Funding, Speakers Bureau; Sumitomo Dainippon Pharma Co., Ltd.: Research Funding, Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Research Funding, Speakers Bureau; AbbVie GK: Research Funding, Speakers Bureau; Pfizer Japan Inc.: Research Funding, Speakers Bureau.

scholarly journals ASXL1 Mutations Predict a Poor Response to Darbepoetin Alfa in Anemic Patients with Low-Risk MDS: A Multicenter, Phase II Study

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 28-29
Author(s):  
Hitoshi Hanamoto ◽  
Yasuyoshi Morita ◽  
Motoshi Ichikawa ◽  
Yasuhito Nannya ◽  
Hirohiko Shibayama ◽  
...  
Keyword(s):  
Board Of Directors ◽  
Odds Ratio ◽  
Research Funding ◽  
Phase Ii Study ◽  
Gene Mutations ◽  
Poor Response ◽  
Low Risk ◽  
Advisory Committees ◽  
Chugai Pharmaceutical ◽  
Otsuka Pharmaceutical

Introduction: Myelodysplastic syndromes (MDS) is thought to develop and progress as a result of accumulation of genetic mutations. This multicenter, open-label, phase II study examined impact of gene mutations on the effect of darbepoetin alfa (DA) for anemia. Death within 1 year and progression to acute myeloid leukemia (AML) were also examined. Methods: DA ≤240 μg was administered once weekly for 16 weeks to DA-naive, low-risk MDS (low or Intermediate-1 [Int-1] risk defined by the International Prognostic Scoring System [IPSS] risk classification) patients with anemia. DNA was extracted from the peripheral blood of patients, and presence of previously reported high-frequency gene mutations in MDS (SF3B1, TET2, SFRS2, ASXL1, DNMT3A, etc.) was analyzed by next-generation sequencing. Primary endpoint was association between frequently observed mutated genes and therapeutic effect of DA (IWG criteria 2006 (HIE)) at 16 weeks. A secondary endpoint was survival analysis results for death and progression to AML within one year after 16 weeks of treatment, with the date of first treatment as the starting date. For AML, progression to AML and subsequent death were stated as events, and the patients without confirmed progression to AML were censored at the date of last known survival. For overall survival, death from any cause was considered an event, and for the survival cases, the study was terminated at the date of last known survival. For the primary endpoint, relative risk and 95% confidence intervals (CI) were calculated using Wilson's score method; statistical significance was assessed using Pearson's chi-square test. Multivariate analysis was performed by adding baseline erythropoietin (EPO) levels (low: &lt;91.8, high: 91.8+ [mIU/mL]) to the explanatory variables, and odds ratio was calculated using logistic regression model. Survival curve was estimated using Kaplan-Meier method and median survival time (MST) was calculated using Brookmeyer and Crowley method. Results: Of the 85 patients enrolled between August 2016 and May 2019, 4 patients who were judged ineligible and 2 patients who discontinued the study prior to the start of treatment were excluded, and 79 subjects were included in the analysis (full analysis set). Of 79 subjects, 52 (65.8%) were male (median age 77.0 [29-90] years). IPSS risk was low in 27 (36.7%) and Int-1 in 50 (63.3%) subjects. The frequently mutated genes (³10%) were SF3B1 (24, 30.4%), TET2 (20, 25.3%), SRSF2 (10 cases, 12.7%), ASXL1 (9, 11.4%), and DNMT3A (8, 10.1%). Overall response rate was 70.9%. Univariate logistic regression analaysis did not show any significant association between these mutations and therapeutic efficacy of DA. The same results were obtained when the analysis was limited to red blood cell transfusion-dependent subjects (n=15). After adjusting against baseline EPO values, mutations of ASXL1 gene were associated with significantly worse response (odds ratio 0.180 [0.035-0.928], p = 0.040). Six (7.6%) confirmed cases of AML and 17 (21.5%) deaths (death before confirmed AML) were observed. Overall, 21 deaths were observed including 4 deaths after progression to AML. The median survival time (95% confidence interval) from the start of treatment to confirmed AML or death was 41.3 months (30.6 months - Not reached). Conclusions: The results of this exploratory study suggest that the presence of ASXL1 gene mutations may result in poor response to the anemic treatment with DA in low-risk MDS. The current analysis of progression to AML and death included subjects whose observation period did not reach the prespecified number of days. For such cases, the outcome research will be conducted around October 2020 and updated results will be presented at the ASH 2020 conference. Disclosures Ichikawa: Novartis, Takeda: Honoraria. Shibayama:Taiho: Research Funding; Shionogi: Research Funding; Ono: Honoraria, Research Funding; Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding; Eisai: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Astellas: Research Funding; Takeda: Honoraria, Research Funding; Merck Sharp & Dohme: Research Funding; Sumitomo Dainippon: Honoraria, Research Funding; Nippon Shinyaku: Honoraria, Research Funding; Daiichi Sankyo: Honoraria; Novartis: Honoraria, Research Funding; Janssen: Honoraria, Research Funding; Chugai: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Kyowa Kirin: Honoraria; Otsuka: Honoraria; Bristol-Myers Squibb: Honoraria; Pfizer: Honoraria; Fujimoto: Honoraria; AbbVie: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; AstraZeneca: Honoraria, Membership on an entity's Board of Directors or advisory committees; Sanofi: Honoraria; Teijin: Research Funding; Mundi Pharma: Honoraria. Maeda:Kyowa Kirin Co.Ltd.: Honoraria, Research Funding. Kawabata:Celgene Corporation: Consultancy; Celgene Corporation: Honoraria; Sanofi K. K: Honoraria; Novartis Pharma K. K.: Honoraria; Chugai Pharmaceutical Co., Ltd: Honoraria. Matsumura:Shionogi & Co., Ltd.: Research Funding; Sumitomo Dainippon Pharma Co., Ltd.: Research Funding; Kyowa Kirin Co., Ltd.: Research Funding; ONO PHARMACEUTICAL CO., LTD.: Research Funding; Eisai Co., Ltd.: Research Funding; Chugai Pharmaceutical Co., Ltd.: Research Funding; Janssen Pharmaceutical K.K: Speakers Bureau; Amgen K.K.: Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Speakers Bureau; Astellas Pharma Inc.: Speakers Bureau; DAIICHI SANKYO COMPANY, LIMITED.: Speakers Bureau; Pfizer Japan Inc.: Speakers Bureau; Novartis Pharma KK: Speakers Bureau; Bristol-Myers Squibb Company: Speakers Bureau. Ogawa:Eisai Co., Ltd.: Research Funding; KAN Research Institute, Inc.: Membership on an entity's Board of Directors or advisory committees, Research Funding; Chordia Therapeutics, Inc.: Membership on an entity's Board of Directors or advisory committees, Research Funding; Asahi Genomics Co., Ltd.: Current equity holder in private company; Sumitomo Dainippon Pharma Co., Ltd.: Research Funding; Otsuka Pharmaceutical Co., Ltd.: Research Funding. Mitani:CHUGAI: Research Funding; Takeda: Research Funding; KYOWA KIRIN: Consultancy, Research Funding.

scholarly journals eIF4B Is a Novel Target of CAMK2G and Promotes Proliferation of Malignant Cells in Myelofibrosis

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 3586-3586
Author(s):  
Ken Sasaki ◽  
Hideaki Mizuno ◽  
Tadayuki Ogawa ◽  
Yosuke Masamoto ◽  
Mineo Kurokawa
Keyword(s):  
Mass Spectrometry ◽  
Pharmaceutical Company ◽  
Research Funding ◽  
Kinase Activity ◽  
Protein Complexes ◽  
Direct Interaction ◽  
Chugai Pharmaceutical ◽  
Company Limited ◽  
Otsuka Pharmaceutical ◽  
Jak2 Inhibitors

Abstract Background Myelofibrosis (MF) is a myeloproliferative neoplasm which is associated with megakaryocytic atypia, fibrosis in the bone marrow (BM), and extramedullary hematopoiesis, followed by progressive hematopoietic failure and leukemic transformation. JAK1/JAK2 inhibitors are currently available and reduce spleen volume, improve symptoms related to MF and prolong the overall survival (OS). Although the benefits associated with JAK1/JAK2 inhibitors are well established, not all patients respond to these inhibitors, and long-term exposure to these inhibitors results in the emergence of resistant clones based on the reactivation of the JAK-STAT pathway. Therefore, new therapeutic strategies targeting other molecules can provide additional benefits to patients with MF. In the previous study, we identified Calcium/Calmodulin Dependent Protein Kinase II Gamma (CAMK2G) as a new therapeutic target for MF by performing compound screening. Furthermore, CAMK2G inhibition can overcome drug-resistance against JAK2 inhibitors. Therefore, it is important to investigate the mechanisms underlying the therapeutic effect of CAMK2G inhibition. In this study, to explore the mechanism underlying the therapeutic effect of CAMK2G inhibition, we performed the immunoprecipitation mass spectrometry to find out the protein that has the direct interaction with CAMK2G. Methods Quantitative Proteomic Analysis of the Target Proteins of CAMK2G The protein complexes with FLAG-tagged CAMK2G (FLAG-CAMK2G) were trapped by anti-FLAG antibody. The eluted assay mixtures were reduced, alkylated and digested into peptides. Each peptide solution and control mixture were labeled with differential stable isotope tags. The samples were quantitatively analyzed using a Q Exactive mass spectrometer (Thermo Fisher Scientific). The spectra were searched against the SWISS-PROT databases using SEQUEST on Proteome Discoverer software 2.2 (Thermo Scientific). Results To reveal the , we conducted the immunoprecipitation assays by FLAG-CAMK2G or FLAG-tag alone, overexpressed in MF model cells. Because CAMK2G has kinase activity, we attempted to reveal the kinase-activity-dependent targets using unhydrolyzable ATP analog, AMP-PNP（5'-adenylyl-imidodiphosphate）that can maintain the strong-binding state of kinase with its target. The protein complexes with FLAG-CAMK2G were trapped by anti-FLAG antibody. After samples were digested into peptides and the candidate proteins were identified and quantitatively analyzed by mass spectrometry. As a result, we identified eukaryotic translation initiation factor 4B (eIF4B) as a protein that has a direct interaction with CAMK2G. eIF4B is a part of the complex involved in the initiation of translation. It has been reported that eIF4B plays a role in the translation of factors involved in anti-apoptosis (Bcl2, Bclxl, Mcl1), cell cycle (Cdc25c), and cell proliferation (c-Myc). We then checked whether knockdown of eIF4B decrease proliferation of MF model cell line. shRNA-mediated silencing of eIF4B decreased cell growth of these cells. Furthermore, the phosphorylation of eIF4B was increased by the ectopic expression of MPL W515L, one of the common mutations found in MF. Also, the phosphorylation of eIF4B was increased by the overexpression of CAMK2G. We then explored the proteins regulated by eIF4B in MF and identified that knockdown of eIF4B decreased the amount of Bcl2 protein. Conclusion In our study, we performed immunoprecipitation mass spectrometry and identified eIF4B as a partner protein of CAMK2G. Since CAMK2G inhibition was shown to be effective against MF in vitro and in vivo, we focused on eIF4B as a potential effector in MF. We also showed that overexpression of MPL W515L and CAMK2G phosphorylates eIF4B and that knockdown of eIF4B inhibited proliferation of MF cells. Furthermore, Bcl2 can be one of the target proteins regulated by eIF4B. Based on these, eIF4B plays an important role in MF. We further perform ribosome profiling to comprehensively understand the regulation of translation by eIF4B. Our research not only elucidate the pathogenesis of MF but also identify new therapeutic targets for MF. Disclosures Masamoto: MSD K.K.: Speakers Bureau; Eisai Co., Ltd.: Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Speakers Bureau; Takeda Pharmaceutical Company Limited.: Speakers Bureau; Chugai Pharmaceutical Company: Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Speakers Bureau; Nippon Shinyaku Co., Ltd.: Speakers Bureau; AbbVie GK: Speakers Bureau; Janssen Pharmaceutical K.K.: Speakers Bureau; SymBio Pharmaceuticals: Speakers Bureau; Bristol Myers Squibb: Speakers Bureau. Kurokawa: Otsuka Pharmaceutical Co., Ltd.: Research Funding, Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Research Funding, Speakers Bureau; Takeda Pharmaceutical Company Limited.: Research Funding, Speakers Bureau; Chugai Pharmaceutical Company: Research Funding, Speakers Bureau; Sumitomo Dainippon Pharma Co., Ltd.: Research Funding, Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Research Funding, Speakers Bureau; Nippon Shinyaku Co., Ltd.: Research Funding, Speakers Bureau; Daiichi Sankyo Company.: Research Funding, Speakers Bureau; AbbVie GK: Research Funding, Speakers Bureau; Teijin Limited: Research Funding, Speakers Bureau; Pfizer Japan Inc.: Research Funding, Speakers Bureau; Eisai Co., Ltd.: Research Funding, Speakers Bureau; MSD K.K.: Research Funding, Speakers Bureau; Astellas Pharma Inc.: Research Funding, Speakers Bureau.

scholarly journals A Nationwide Survey of Lower Risk Myelodysplastic Syndromes in Japan

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 4662-4662
Author(s):  
Kumi Nakazaki ◽  
Toshiaki Takezaki ◽  
Yosuke Masamoto ◽  
Yasushi Miyazaki ◽  
Kinuko Mitani ◽  
...  
Keyword(s):  
Pharmaceutical Company ◽  
Lower Risk ◽  
Research Funding ◽  
Anabolic Steroids ◽  
Risk Groups ◽  
Free Survival ◽  
Chugai Pharmaceutical ◽  
Company Limited ◽  
Otsuka Pharmaceutical ◽  
Research Facilities

Abstract Introduction: There are some treatment options for lower risk myelodysplastic syndromes (MDS) such as erythropoiesis-stimulating agents (ESAs), anabolic steroids, hypomethylating agents, and immunosuppressants. The object of this multicenter retrospective study was to survey the current situation about treatment selection and the prognosis of the lower risk MDS cases in Japan. We also evaluated the prognosis of the cases with paroxysmal nocturnal hemoglobinuria (PNH) type cells and therapeutic effects of cyclosporine. Methods: We investigated the clinical information in the form of a questionnaire for joint research facilities as to each case of newly diagnosed MDS between 2013 and 2018 corresponding to the lower risk of International Prognostic Scoring System (IPSS) or revised IPSS (IPSS-R). The diagnosis of MDS was based on WHO 2008 or WHO 2016 classification. Survival analysis was conducted using Kaplan-Meier method and log-rank test. Multivariate analysis was performed using Cox proportional hazard regression model. This study was approved by the institutional review board of the University of Tokyo and other research facilities. This work was supported by the Research Program of Intractable Disease (the Japanese National Research Group on Idiopathic Bone Marrow Failure Syndromes) provided by the Ministry of Health, Labor, and Welfare of Japan. Results: 1,304 cases at thirty facilities nationwide were enrolled. Median age was 76 years [IQR, 68 - 83], and male and female ratio was 61.3% and 38.7%. At diagnosis, 19.0% and 4.4% of the cases were dependent on red blood cells and platelets transfusion, respectively. The risk classification of enrolled cases was as follows: very low, 217 (16.6%); low, 652 (50.0%); intermediate, 360 (27.6%); high, 56 (4.3%); very high, 4 (0.3%); not determined, 14 (1.1%). 1,230 cases of the very low, low and intermediate risk groups were included in subsequent analyzes. Serum erythropoietin levels were measured in 466 cases (37.9%) with a median of 61.8 IU/l, and 74.2% and 85.7% cases showed less than 200 IU/l and 500 IU/l, respectively. PNH type cells in the peripheral blood were evaluated in 231 cases and were positive in 33 cases (14.3%). Median follow-up period was 22 months. As an initial therapy, 26.4% and 11.6% of transfusion-dependent and independent cases started to receive ESAs, respectively. 16.6% and 11.5% took oral anabolic hormones, and azacytidine were administered to 17.0% and 7.2% of each group. 55.4% of transfusion-independent cases were just followed up at first. Median overall and acute myeloid leukemia (AML)-free survival was 70.0 months [95% CI, 61.0 - not reached] and 62.0 months [95% CI, 54.0 - 74.0], respectively. Log-rank analysis revealed significant differences among IPSS-R risk groups about overall and AML-free survival (p&lt;0.01 and p&lt;0.01, respectively). Multivariate analysis confirmed that initiating azacytidine at the time of diagnosis conferred an independent significant poor prognostic factor with respect to overall survival (hazard ratio for death, 1.74; p&lt;0.01) and AML-free survival (hazard ratio for death or onset of AML, 1.86; p&lt;0.01) in addition to sex, age, IPSS-R classification, and transfusion-dependency. Comparing thirty-three positive cases of PNH type cells with 198 negative cases, overall and AML-free survival was significantly better in the former group (p&lt;0.01 and p&lt;0.01, respectively). Median overall and AML-free survival were not reached in the positive group and 51.0 months [95% CI, 47.0 - 79.0] and 49.1 months [95% CI, 40.0 - 57.1] in the negative group, respectively. Interestingly, focused on the positive cases, 14 cases receiving cyclosporine revealed better AML-free survival than the others (p=0.033). Overall survival was tended to be better (p=0.052). On the other hand, cyclosporine did not improve the prognosis of the PNH type cells negative cases. It is thought to be consistent with the effectiveness of immunosuppressive therapy in aplastic anemia with PNH type cells. Conclusion: ESAs, anabolic steroids, and azacytidine were frequently selected as an initial treatment for lower risk MDS cases in Japan, however, when to start azacytidine is an issue for consideration. Good response to cyclosporine may be obtained in cases with PNH type cells. Disclosures Masamoto: Eisai Co., Ltd.: Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Speakers Bureau; Takeda Pharmaceutical Company Limited.: Speakers Bureau; Chugai Pharmaceutical Company: Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Speakers Bureau; Nippon Shinyaku Co., Ltd.: Speakers Bureau; AbbVie GK: Speakers Bureau; Janssen Pharmaceutical K.K.: Speakers Bureau; SymBio Pharmaceuticals: Speakers Bureau; Bristol Myers Squibb: Speakers Bureau; MSD K.K.: Speakers Bureau. Miyazaki: Kyowa-Kirin: Honoraria; Sumitomo-Dainippon: Honoraria, Research Funding; Abbvie: Honoraria; Novartis: Honoraria; Nippon-Shinyaku: Honoraria; Bristol-Myers Squibb: Honoraria; Pfizer: Honoraria; Astellas: Honoraria; Janssen: Honoraria; Eisai: Honoraria; Daiichi-Sankyo: Honoraria; Chugai: Honoraria; Takeda: Honoraria; Sanofi: Honoraria. Mitani: Nippon Shinyaku Co.: Research Funding, Speakers Bureau; MSD Pharma.: Research Funding, Speakers Bureau; Novartis: Speakers Bureau; Pfizer Inc.: Speakers Bureau; Celgene Co.: Speakers Bureau; Takeda Pharma.: Research Funding, Speakers Bureau; Kyowa Kirin,: Research Funding, Speakers Bureau; Bristol Myers Squibb: Speakers Bureau; Shire plc: Speakers Bureau; BML Inc: Speakers Bureau; Mochida Parma.: Speakers Bureau; Alexion Pharma.: Speakers Bureau; AbbVie Inc.: Speakers Bureau; Ono Pharma.: Speakers Bureau; Chugai Pharma.: Research Funding; Teijin Pharma.: Research Funding; Sumitomo Dainippon Pharma: Research Funding; Taiho Phama.: Research Funding; Otsuka Pharma.: Research Funding. Kurokawa: Sumitomo Dainippon Pharma Co., Ltd.: Research Funding, Speakers Bureau; AbbVie GK: Research Funding, Speakers Bureau; Teijin Limited: Research Funding, Speakers Bureau; Pfizer Japan Inc.: Research Funding, Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Research Funding, Speakers Bureau; Takeda Pharmaceutical Company Limited.: Research Funding, Speakers Bureau; Daiichi Sankyo Company.: Research Funding, Speakers Bureau; Eisai Co., Ltd.: Research Funding, Speakers Bureau; Nippon Shinyaku Co., Ltd.: Research Funding, Speakers Bureau; MSD K.K.: Research Funding, Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Research Funding, Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Research Funding, Speakers Bureau; Chugai Pharmaceutical Company: Research Funding, Speakers Bureau; Astellas Pharma Inc.: Research Funding, Speakers Bureau.

scholarly journals Comparison of Minimal Residual Disease Detection between the Manual and Automated Gating Strategies of Euroflow Next-Generation Flow in Patients with Multiple Myeloma

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 3083-3083
Author(s):  
Hiroyuki Takamatsu ◽  
Takeshi Yoroidaka ◽  
Takeshi Yamashita ◽  
Ryoichi Murata ◽  
Mikio Ueda ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Partial Response ◽  
Pharmaceutical Company ◽  
Research Funding ◽  
Plasma Cells ◽  
Residual Disease ◽  
Next Generation ◽  
Chugai Pharmaceutical ◽  
Company Limited ◽  
Minimal Residual

Background: The rate of complete response (CR) in multiple myeloma (MM) has dramatically increased because of the development of novel agents. In addition, the development of methods for measuring minimal residual disease (MRD), such as multiparameter flow cytometry and next-generation sequencing, has made it possible to stratify CR patients according to the MRD extent. EuroFlow next-generation flow (EuroFlow-NGF) is considered one of the gold standard methods for evaluating the negative status of MRD in MM. The automated gating strategy of EuroFlow-NGF has been shown to detect MRD as accurately as the manual gating strategy by experts. Oberle et al. (Haematologica, 2017) have found that daratumumab persisted on the surface of myeloma cells treated with it and that the anti-CD38 multi-epitope antibody used in EuroFlow-NGF has partial cross-reactivity with daratumumab, leading to generally lower mean fluorescence intensities of CD38. Therefore, MRD levels may have been underestimated in patients who were treated with anti-CD38 monoclonal antibodies (mAbs) using the automated gating strategy, leading to inappropriate management of the patients. Because no studies have examined the correlation of MRD extent between the manual and automated gating strategies in patients with MM who have received anti-CD38 mAbs, we compared MRD detection between the two gating strategies of EuroFlow-NGF in patients with MM. Methods: The study included bone marrow samples from 51 patients with MM (27 male and 24 female patients), including 13 patients treated with anti-CD38 mAb (12 treated with daratumumab and 1 treated with isatuximab). The median patient age was 70 years (range, 32-92 years) at MRD assessment. The disease statuses at MRD assessment were stringent CR in 26 patients (51%), CR in 7 (14%), very good partial response in 13 (26%), partial response in 1 (2%), and progressive disease in 4 (8%). The sample preparation protocol, Ab panel, and automated gating strategy of EuroFlow-NGF have been reported previously (Flores-Montero et al. Leukemia 2017). Briefly, we performed the EuroFlow-NGF method, which involved ammonium chloride-based bulk lysis, followed by surface staining using antibodies against CD138-BV421, CD27-BV510, CD38 multiepitope (ME)-FITC, CD56-PE, CD45-PerCP Cy5.5, CD19-PECy7, CD117-APC, and CD81-APC C750 in tube 1 and surface/intracellular staining using antibodies against CD138-BV421, CD27-BV510, CD38 ME-FITC, CD56-PE, CD45-PerCP Cy5.5, CD19-PECy7, CD117-APC, CD81-APC C750, cytoplasmic (cy) Igκ-APC, and cyIgλ-APC C750 after permeabilization in tube 2. For data analysis, events from both eight-color tubes (tubes 1 and 2) were merged, and the values of all parameters per tube were mathematically calculated using the merge and calculation functions of Infinicyt software (Cytognos SL, Salamanca, Spain). Automatic identification and enumeration of total plasma cells (tPCs) and abnormal plasma cells (MRD) were performed using the automatic gating function of Infinicyt software as described previously (Flores-Montero et al. Leukemia 2017). We compared the total nucleated cell number, tPC ratio, and MRD ratio between the manual (by experts) and automated gating strategies of EuroFlow-NGF. Results: In patients with MM who did not receive any anti-CD38 mAb therapy, we observed high correlations for both the tPC (r = 0.959, P < 0.0001) (Figure A) and MRD (r = 0.974, P < 0.0001) (Figure B) ratios between the manual and automated gating strategies of EuroFlow-NGF. On the other hand, in patients with MM who received anti-CD38 mAb therapy, we did not observe good correlations for both the tPC (r = 0.349, P = 0.2) (Figure A) and MRD (r = 0.292, P = 0.3) (Figure B) ratios between the two strategies owing to a lower fluorescence intensity of CD38 on PCs. In addition, when the MRD threshold was set to 10-5, the discordance of MRD positivity/negativity between the two strategies was significantly higher in patients who received anti-CD38 mAb therapy than in those who did not receive anti-CD38 mAb therapy [4/13 (31%) vs. 1/38 (3%), P = 0.012]. Conclusion: Although the automated gating strategy of EuroFlow-NGF could be a viable alternative to the manual strategy for the assessment of MRD in MM, we may have to utilize the manual strategy to obtain precise MRD results for patients with MM who received anti-CD38 mAbs. Figure Disclosures Takamatsu: Celgene: Consultancy, Honoraria, Research Funding; Bristol-Myers Squibb: Honoraria, Research Funding; Ono pharmaceutical: Honoraria, Research Funding; CSL Behring: Research Funding; SRL: Consultancy, Research Funding; Janssen Pharmaceutical: Consultancy, Honoraria; Sanofi: Consultancy, Honoraria; Takeda Pharmaceutical Company Limited: Honoraria; Fujimoto Pharmaceutical: Honoraria; Becton, Dickinson and Company: Honoraria; Abbvie: Consultancy; Daiichi-Sankyo Company: Honoraria. Yoroidaka:Ono Pharmaceutical: Honoraria. Yamashita:Janssen Pharmaceutical K.K.: Honoraria; Daiichi-Sankyo Company: Honoraria; Kyowa Kirin: Honoraria; Chugai Pharmaceutical Co.,Ltd: Honoraria; TEIJIN PHARMA LIMITED: Honoraria; Takeda Pharmaceutical Company Limited: Honoraria; Bristol-Myers Squibb: Honoraria; Ono Pharmaceutical: Honoraria; Celgene: Honoraria. Murata:Celgene: Honoraria; Ono pharmaceutical: Honoraria. Nakao:Daiichi-Sankyo Company, Limited: Honoraria; Janssen Pharmaceutical K.K.: Honoraria; SynBio Pharmaceuticals: Consultancy; Ohtsuka Pharmaceutical: Honoraria; Celgene: Honoraria; Ono Pharmaceutical: Honoraria; Novartis Pharma K.K: Honoraria; Bristol-Myers Squibb: Honoraria; Takeda Pharmaceutical Company Limited: Honoraria; Chugai Pharmaceutical Co.,Ltd: Honoraria; Kyowa Kirin: Honoraria; Alaxion Pharmaceuticals: Honoraria. Matsue:Novartis Pharma K.K: Honoraria; Ono Pharmaceutical: Honoraria; Takeda Pharmaceutical Company Limited: Honoraria; Celgene: Honoraria; Janssen Pharmaceutical K.K.: Honoraria.

scholarly journals Intrapatient Complex Transcriptional Responses to Conventional Chemotherapy in Relapsed Acute Myeloid Leukemia Revealed By Single Cell RNA-Seq Analysis

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 1179-1179
Author(s):  
Hideaki Mizuno ◽  
Akira Honda ◽  
Mineo Kurokawa
Keyword(s):  
Single Cell ◽  
Signaling Pathway ◽  
Myeloid Leukemia ◽  
Research Funding ◽  
Chemotherapy Resistance ◽  
Rna Seq ◽  
Conventional Chemotherapy ◽  
Chugai Pharmaceutical ◽  
Dna Mutation ◽  
Otsuka Pharmaceutical

Abstract Resistance to anthracycline and cytarabine based conventional chemotherapy often occurs and results in extremely poor prognosis in patients with acute myeloid leukemia (AML). Although chemotherapy resistance is the most critical clinical problem, the mechanisms by which AML confers resistance to conventional chemotherapy are not yet fully understood. In this study, we investigated the key mechanisms of chemotherapy resistance through single cell RNA-sequencing analysis using paired bone marrow AML cells longitudinally collected from two AML-MRC patients at diagnosis and relapse after anthracycline-based chemotherapy. AML blasts were sorted by CD45/SSC gating and subjected to single cell RNA-seq analysis. Single cell RNA-seq was performed using 10x Genomics' Chromium System. Mean estimated number of cells per sample was 3.403 (2,731-4,200) and median detected genes per cell ranged 3,030 to 3,918 among four samples. Data collected from paired samples were combined in following analysis. Transcriptome based clustering following UMAP dimensionality reduction distinguished 5 and 9 cluster groups in each paired sample. Chemotherapy sensitive cluster groups dominant at diagnosis and chemotherapy resistant cluster groups dominant at relapse were clearly divided. In each paired sample, a few AML cells at diagnosis were allocated to chemotherapy resistant cluster groups. This suggested that transcriptionally identifiable less frequent cells resistant to chemotherapy existed at diagnosis and may expand during and/or after chemotherapy maintaining its transcriptional features. Next, to determine whether these transcriptional features are correlated with DNA mutation profiles, we labeled DNA mutation status to each cell and compared frequencies of mutation. As far as we detected, AML recurrent mutations such as DNMT3A R882C and TP53 missense mutation were not related to chemotherapy resistant cluster groups, although this method was relatively limited by the nature of RNA-seq-based mutation detection. Then we sought to determine transcriptional features of resistant clones. Gene set enrichment analysis identified some gene groups such as E2F signaling pathway, MYC signaling pathway, hedgehog signaling pathway and TNFA signaling pathway as transcriptional signatures related to emergence after chemotherapy. Analysis of known hematopoietic differentiation gene signatures showed distinct differentiation profiles in each cluster groups, whereas resistant cluster groups were not necessarily related to hematopoietic stem cell signatures. Intrapatient variations of transcriptional signatures among the resistant cluster groups were detected, which indicated that accurate detection of transcriptional features related to chemotherapy resistance may be difficult by using bulk RNA-seq method. As for other cluster groups which were not dominant both at diagnosis and relapse, these cluster groups hardly changed its frequencies between at diagnosis and relapse, which suggested less proliferative leukemia cells persisted during chemotherapy and have various transcriptional features although whether these persisting cells contribute to relapse was unclear. Since enriched transcriptional signatures in resistant cluster groups were not consistent between the two patients, further analysis using samples collected from more patients would be needed to determine common critical chemotherapy resistant transcriptional signature. In conclusion, our analysis suggested that a transcriptionally identifiable small fraction of cells showing gene signatures related to chemotherapy resistance at diagnosis may expand during chemotherapy and revealed intrapatient transcriptional complexity of response to chemotherapy, which cannot be uncovered by bulk RNA-sequencing. Disclosures Honda: Takeda Pharmaceutical: Other: Lecture fee; Otsuka Pharmaceutical: Other: Lecture fee; Chugai Pharmaceutical: Other: Lecture fee; Ono Pharmaceutical: Other: Lecture fee; Jansen Pharmaceutical: Other: Lecture fee; Nippon Shinyaku: Other: Lecture fee. Kurokawa: MSD K.K.: Research Funding, Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Research Funding, Speakers Bureau; Daiichi Sankyo Company.: Research Funding, Speakers Bureau; Astellas Pharma Inc.: Research Funding, Speakers Bureau; Pfizer Japan Inc.: Research Funding, Speakers Bureau; Nippon Shinyaku Co., Ltd.: Research Funding, Speakers Bureau; Sumitomo Dainippon Pharma Co., Ltd.: Research Funding, Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Research Funding, Speakers Bureau; Eisai Co., Ltd.: Research Funding, Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Research Funding, Speakers Bureau; Teijin Limited: Research Funding, Speakers Bureau; Takeda Pharmaceutical Company Limited.: Research Funding, Speakers Bureau; Chugai Pharmaceutical Company: Research Funding, Speakers Bureau; AbbVie GK: Research Funding, Speakers Bureau.

scholarly journals EVI1-Positive AML Cells Show Distinct Features and High Dependency on ETS Transcription Factor ERG

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 784-784
Author(s):  
Yosuke Masamoto ◽  
Akira Chiba ◽  
Toshiaki Takezaki ◽  
Toshiya Hino ◽  
Hiroki Hayashida ◽  
...  
Keyword(s):  
Pharmaceutical Company ◽  
Progenitor Cells ◽  
Research Funding ◽  
Cytotoxic Agents ◽  
Hematopoietic Progenitor Cells ◽  
Hematopoietic Progenitor ◽  
Downstream Target ◽  
Company Limited ◽  
Otsuka Pharmaceutical ◽  
Ets Transcription Factor

Abstract Inappropriate expression of Ecotropic viral integration site 1 (EVI1) has been associated with dismal clinical outcomes in acute myeloid leukemia (AML), while EVI1 is indispensable for normal hematopoiesis. We have previously reported that EVI1 expression is restricted to hematopoietic stem cell fraction and EVI1-expressing cells show robust long-term reconstitution capacity using Evi1-IRES-GFP knock-in (EVI1-GFP) mice, which enable us to track Evi1 expression on a single cell basis. In this study, we tried to elucidate the functional implication of EVI1 expression in AML using these mice. We generated murine EVI1-GFP AML model by retrovirally transducing MLL-AF9 or -ENL fusion gene into Lineage- Sca-1+ c-kit+ (LSK) cells from EVI1-GFP mice followed by transplantation into lethally irradiated syngeneic mice. Clonogenic and leukemogenic potentials of AML cells, especially leukemic cells with a granulocyte-macrophage progenitor phenotype (L-GMPs) from these mice, were compared according to GFP expression. Remarkably, GFP-positive L-GMPs tended to show lower colony-forming activity in semi-solid media and lower leukemia-initiating potential than GFP-negative L-GMPs. GFP-positive L-GMPs, however, induced a more aggressive form of AML, characterized by shorter survival in the secondary transplantation model. When EVI1-GFP AML mice underwent cytotoxic chemotherapy with cytarabine, the GFP-positive fraction was enriched during myelosuppression, indicating the survival advantage of EVI1-positive cells. To investigate the downstream target of EVI1, we employed murine EVI1-AML model, where murine hematopoietic cells exogenously expressing 3×FLAG-tagged EVI1 were transplanted into syngeneic mice. Using EVI1-AML cells, we performed chromatin-immunoprecipitation coupled to next-generation sequencing (ChIP-seq) by anti-FLAG tag antibody. To identify leukemia-specific targets of EVI1, the result was compared with the result of ChIP-seq obtained from 32D-cl3 murine hematopoietic progenitor cells with 3×FLAG tag inserted into 3'-end of the coding region of the EVI1 gene. Gene ontology analysis revealed that genes involved in immune processes are explicitly enriched in the leukemia samples. Among the list of EVI1-bound genes, we tried to refine functional downstream targets of EVI1, which are upregulated in murine EVI1-AML cells and of which expressions are positively correlated with EVI1. By combining the ChIP-seq data with murine transcriptome data that compare hematopoietic progenitor cells expressing empty-vector and EVI1+ AML cells, and public gene expression datasets of human AML (Valk et al. NEJM 2004), we picked out 18 genes as candidate EVI1 downstream genes. Functional screening using EVI1-AML cells and shRNAs against these genes revealed that silencing of ETS transcription factor ERG (ETS-related gene) markedly suppressed proliferation and colony-forming activity of EVI1-AML cells, as well as rendered them vulnerable to cytotoxic agents. Normal c-kit+ hematopoietic progenitor cells were less affected by shRNAs against ERG. By comparing MLL-ENL immortalized murine hematopoietic cells with high and low EVI1 expression, EVI1-high MLL-ENL cells showed higher ERG dependency than EVI1-low MLL-ENL cells. Pharmacological inhibition of ERG also led to marked inhibition of EVI1-AML cells and EVI1-high MLL-ENL cells. Finally, knockdown of ERG remarkably delayed AML development in bone marrow transplantation model of EVI1-AML and EVI1-expressing MLL-ENL AML. Our data suggest that EVI1-positive AML cells are characterized by an aggressive nature and resistance to cytotoxic agents, as well as low leukemia stem cell capacity. ERG would be a novel downstream target of EVI1, on which survival of EVI1-expressing AML cells depends. Disclosures Masamoto: Kyowa Hakko Kirin Co., Ltd.: Speakers Bureau; Chugai Pharmaceutical Company: Speakers Bureau; Bristol Myers Squibb: Speakers Bureau; Janssen Pharmaceutical K.K.: Speakers Bureau; Eisai Co., Ltd.: Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Speakers Bureau; MSD K.K.: Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Speakers Bureau; Takeda Pharmaceutical Company Limited.: Speakers Bureau; Nippon Shinyaku Co., Ltd.: Speakers Bureau; AbbVie GK: Speakers Bureau; SymBio Pharmaceuticals: Speakers Bureau. Kurokawa: Daiichi Sankyo Company.: Research Funding, Speakers Bureau; Eisai Co., Ltd.: Research Funding, Speakers Bureau; MSD K.K.: Research Funding, Speakers Bureau; Astellas Pharma Inc.: Research Funding, Speakers Bureau; Teijin Limited: Research Funding, Speakers Bureau; Nippon Shinyaku Co., Ltd.: Research Funding, Speakers Bureau; ONO PHARMACEUTICAL CO., LTD.: Research Funding, Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Research Funding, Speakers Bureau; AbbVie GK: Research Funding, Speakers Bureau; Pfizer Japan Inc.: Research Funding, Speakers Bureau; Chugai Pharmaceutical Company: Research Funding, Speakers Bureau; Otsuka Pharmaceutical Co., Ltd.: Research Funding, Speakers Bureau; Takeda Pharmaceutical Company Limited.: Research Funding, Speakers Bureau; Sumitomo Dainippon Pharma Co., Ltd.: Research Funding, Speakers Bureau.

scholarly journals The Outcomes of Systemic Chronic Active EBV Infection Treatment By Allogeneic Hematopoietic Stem Cell Transplantation: An Analysis of Japanese Registry Data

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 3965-3965
Author(s):  
Ayako Arai ◽  
Masahide Yamamoto ◽  
Maho Sato ◽  
Yasushi Onishi ◽  
Yoji Sasahara ◽  
...  
Keyword(s):  
Stem Cell ◽  
Stem Cell Transplantation ◽  
Cell Transplantation ◽  
Research Funding ◽  
Skin Lesions ◽  
Hematopoietic Stem ◽  
Chugai Pharmaceutical ◽  
Ebv Infection ◽  
Otsuka Pharmaceutical ◽  
Definition Of

Abstract Background and aims Systemic chronic active Epstein-Barr virus infection (sCAEBV) is classified as T- or NK-cell neoplasms in the WHO classification revised in 2017. Although allogeneic stem cell transplantation (allo-HSCT) is efficacious for sCAEBV, the effects are yet to be analyzed in a large number of cases due to the disease rarity. To investigate the outcomes and the prognostic factors of allo-HSCT in sCAEBV under the definition of the WHO 2017 classification, we analyzed retrospectively using the database of Japanese Society for Transplantation and Cellular Therapy (JSTCT). Methods Data collection We used the clinical data of hematopoietic stem cell transplantation (HSCT) recipients of the Transplant Registry Unified Management Program (TRUMP) sponsored by JSTCT and Japanese Data Center for Hematopoietic Cell Transplantation (JDCHCT). Patients who underwent HSCT to cure EBV-associated diseases, secondary hemophagocytic lymphohistiocytosis (HLH), and virus-associated hemophagocytic syndrome between January 1993 and December 2016 were selected in TRUMP database, and on our behalf, JDCHCT sent out survey questions to the institutions of these patients to collect additional data to check if the diagnosis of sCAEBV matches our criteria, to analyze disease status at the time of HSCT, and to evaluate the efficacy of different treatment methods. The diagnosis of sCAEBV sCAEBV was diagnosed according to criteria suggested in 2016 by the Research group of Measures against Intractable Diseases by Ministry of Health, Labour and Welfare of Japan: (1) elevated EBV DNA load in peripheral blood (PB) (&gt; 10 2.5 copies/μg DNA), (2) detection of EBV infection in T or NK cells from the affected tissues or PB, (3) systemic inflammatory symptoms such as fever, lymphadenopathy, liver dysfunction, progressive skin lesions, vasculitis, and uveitis persisting for &gt; 3 months, and (4) exclusion of other possible diseases, such as primary EBV infection, autoimmune disease, immunodeficiencies, and lymphomas. Patients who fulfilled all (1) to (4) were diagnosed as sCAEBV. These criteria are compatible with the definition of sCAEBV described in the WHO definition of 2017. The definitions of disease activities and responses The disease activities are defined in previous reports (Blood. 2012;119, p673 and BMT. 2016;51, p879) as follows: positive of fever, ALT level elevation, vasculitis, progressive skin lesions, or uveitis. We defined the complete resolution of disease activity as complete response (CR) and CR with a significant decrease in PB EBV-DNA load (&lt; 10 2.5 copies/μg DNA) as virological CR (vCR). Results 81 patients who met the diagnostic criteria of sCAEBV were analyzed. The median age at HSCT was 24 years old, and the three-year overall survival rate (3-year OS) was 74.0%. Of 74 patients whose viral load after HSCT evaluated, 49 (66.2%) achieved vCR. The multivariate cox proportional hazard model revealed that advanced age, adolescent and young adult (AYA) (age, 15-39; n = 48) and adult (age, &gt; 40; n = 13), was a risk factor of poor OS. The hazard ratios (HR) of AYA and adult groups were 10.14 and 4.63 respectively. It also showed that the presence of HLH at HSCT (HR 4.55), high sIL-2R (≥ median, 691 U/mL) at HSCT (HR 5.27), and conditioning without total body irradiation (HR 3.23) were independently associated with poor survival. Moreover, the median survival time of patients with active disease and extremely high sIL-2R level (≥ 3 × median, 2073 U/mL) was 0.9 months, whereas the other groups did not reach the median. Conclusion Although HSCT is the only curative treatment for sCAEBV, the strategies need improvements in high-risk cases, especially of high sIL-2R. Disclosures Arai: ONO PHARMACEUTICAL CO., LTD.: Honoraria, Research Funding; CHUGAI PHARMACEUTICAL CO., LTD.: Honoraria, Research Funding; Kyowa Kirin CO., LTD.: Honoraria, Research Funding; Abbvie: Honoraria; BMS: Honoraria; Elsai Co Ltd: Research Funding; Abbott Japan LLC: Honoraria; Nippon Shinyaku Co. Ltd: Honoraria, Research Funding; Otsuka Pharmaceuticals Co. Ltd: Research Funding; Novartis Pharma KK: Honoraria; Takeda Pharmaceuticals Co Ltd: Honoraria, Research Funding; Shionogi & Co Ltd: Research Funding; Asahi Kasri Pharma Corporation: Research Funding; Sanofi: Honoraria; Pfizer japan: Honoraria; Astellas Pharma Inc.: Honoraria. Yamamoto: Bristol-Myers Squibb Company: Honoraria; Chugai Pharmaceutical Co., Ltd.: Honoraria; Eisai Co., Ltd.: Honoraria; Kyowa Kirin Co., Ltd.: Honoraria; NIPPON SINYAKU CO., LTD: Honoraria; Novartis Pharma: Honoraria; ONO PHARMACEUTICAL CO.: Honoraria; Otsuka Pharmaceutical: Honoraria; Pfizer Japan Inc.: Honoraria; Takeda: Honoraria. Nakamae: Astellas Pharma Inc.: Honoraria; Otsuka Pharmaceutical Co., Ltd: Honoraria; ONO PHARMACEUTICAL CO., LTD.: Honoraria; Simon-Kucher & Partners: Honoraria; Sumitomo Dainippon Pharma Co., Ltd.: Honoraria; Takeda Pharmaceutical Company Limited.: Honoraria; Novartis: Honoraria, Research Funding; Pfizer Japan Inc.: Honoraria; Bristol-Myers Squibb　Company: Honoraria, Research Funding; Alexion: Research Funding; PPD-SNBL K.K: Research Funding; CMIC HOLDINGS Co., Ltd: Research Funding. Ichinohe: Repertoire Genesis Inc.: Honoraria, Research Funding; Novartis Pharma K.K.: Honoraria; Celgene: Honoraria; Zenyaku Kogyo Co.: Research Funding; Takara Bio Inc.: Research Funding; Taiho Pharmaceutical Co.: Research Funding; Sumitomo Dainippon Pharma Co.: Honoraria, Research Funding; Otsuka Pharmaceutical Co.: Research Funding; Nippon Shinyaku Co: Research Funding; Ono Pharmaceutical Co.: Honoraria, Research Funding; Kyowa Kirin Co.: Honoraria, Research Funding; FUJIFILM Wako Chemicals.: Honoraria, Research Funding; Daiichi Sankyo: Research Funding; Eisai Co.: Honoraria, Research Funding; CSL Behring: Honoraria, Research Funding; Chugai Pharmaceutical: Research Funding; Bristol-Myers Squibb: Honoraria; Takeda Pharmaceutical Co.: Honoraria; AbbVie Pharma: Research Funding; Astellas Pharma: Honoraria, Research Funding. Atsuta: Astellas Pharma Inc.: Speakers Bureau; Mochida Pharmaceutical Co., Ltd.: Speakers Bureau; AbbVie GK: Speakers Bureau; Kyowa Kirin Co., Ltd: Honoraria; Meiji Seika Pharma Co, Ltd.: Honoraria.

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