Agency For Medical Research And Development Research Articles

Seroprevalence of Bas-Congo virus in Mangala, Democratic Republic of the Congo: a population-based cross-sectional study

Bas-Congo virus (BASV), an emerging tibrovirus, was associated with an outbreak of acute haemorrhagic fever in Mangala, Democratic Republic of the Congo, in 2009. In 2012, neutralising antibodies to BASV were detected in the lone survivor and one of his close contacts. However, subsequent serological and molecular surveys were unsuccessful as neither BASV antibodies nor its RNA were detected. In this study, we determined the seroprevalence of BASV infection in Mangala 13 years after the initial outbreak. We conducted a population-based serological survey from Jan 17 to Jan 23, 2022. Consenting individuals at least 5 years of age, living in Mangala for at least 4 weeks, and who had no contraindication to venepuncture were enrolled. Participants were interviewed using a pre-tested questionnaire for sociodemographic and clinical characteristics. We supplemented the collected serum samples with 284 archived samples from Matadi and Kinshasa. All samples were tested for antibodies to BASV and other tibroviruses using a pseudovirus-based neutralisation test. Among the 267 individuals from Mangala, the prevalence of BASV antibodies was 55% (95% CI 49-61; n=147). BASV seropositivity odds significantly increased with age (5·2 [95% CI 2·1-12·9] to 83·9 [20·8-337·7] times higher in participants aged 20 years or older than participants aged 5-19 years). Some occupational categories (eg,farmer or public servant) were associated with seropositivity. Only nine (6%) of 160 samples from Matadi and one (<1%) of 124 samples from Kinshasa had neutralising antibodies to BASV. Moreover, we also detected neutralising antibodies to other tibroviruses-Ekpoma virus 1, Ekpoma virus 2, and Mundri virus-in 84 (31%), 251 (94%), and 219 (82%) of 267 Mangala samples; 14 (9%), 62 (39%), and 120 (75%) of 160 Matadi samples; and six (5%), five (4%), and 33 (27%) of 124 Kinshasa samples, respectively. Human infection with BASV and other tibroviruses seems common in Mangala, although no deadly outbreak has been reported since 2009. Exposure to BASV might be highly restricted to Mangala and the increasing prevalence of neutralising antibodies with age suggests regular contact with the virus in this city. Altogether, our findings suggest that human infection with tibroviruses could be common in the study areas and not associated with deadly haemorrhagic or debilitating syndromes. Japan Agency for Medical Research and Development (AMED) and Japan International Cooperation Agency (JICA) under the Science and Technology Research Partnership for Sustainable Development (SATREPS) and Japan Program for Infectious Diseases Research and Infrastructure from AMED.

Characterization of a human H3N8 influenza virus

In 2022 and 2023, novel reassortant H3N8 influenza viruses infected three people, marking the first human infections with viruses of this subtype. Here, we generated one of these viruses (A/Henan/4-10CNIC/2022; hereafter called A/Henan/2022 virus) by using reverse genetics and characterized it. In intranasally infected mice, reverse genetics-generated A/Henan/2022 virus caused weight loss in all five animals (one of which had to be euthanized) and replicated efficiently in the respiratory tract. Intranasal infection of ferrets resulted in minor weight loss and moderate fever but no mortality. Reverse genetics-generated A/Henan/2022 virus replicated efficiently in the upper respiratory tract of ferrets but was not detected in the lungs. Virus transmission via respiratory droplets occurred in one of four pairs of ferrets. Deep-sequencing of nasal swab samples from inoculated and exposed ferrets revealed sequence polymorphisms in the haemagglutinin protein that may affect receptor-binding specificity. We also tested 90 human sera for neutralizing antibodies against reverse genetics-generated A/Henan/2022 virus and found that some of them possessed neutralizing antibody titres, especially sera from older donors with likely exposure to earlier human H3N2 viruses. Our data demonstrate that reverse genetics-generated A/Henan/2022 virus is a low pathogenic influenza virus (of avian influenza virus descent) with some antigenic resemblance to older human H3N2 viruses and limited respiratory droplet transmissibility in ferrets. This work was supported by the Japan Program for Infectious Diseases Research and Infrastructure (JP23wm0125002), and the Japan Initiative for World-leading Vaccine Research and Development Centers (JP233fa627001) from the Japan Agency for Medical Research and Development (AMED).

Recent advances in hepatitis A virus research and clinical practice guidelines for hepatitis A virus infection in Japan.

In 2018, there was a hepatitis A outbreak in Japan, and hepatitis A virus (HAV) infection is considered a sexually transmitted disease. In general, patients with hepatitis A should be given attention, and this disease should be prevented more than ever. The Japan Agency for Medical Research and Development (AMED) Hepatitis A and E viruses (HAV and HEV) Study Group has worked on the project to create "Recent Advances in Hepatitis A Virus (HAV) Research and Clinical Practice Guidelines for HAV Infection in Japan". The group consists of expert hepatologists and virologists who gathered at virtual meeting on August 5, 2023. Data about the pathogenesis, infection routes, diagnosis, complications, several factors for the severities, vaccination, and current and future treatments for hepatitis A were discussed and debated for a draft version. The participants assessed the quality of cited studies. The finalized recommendations are presented in this review. The recent advances in HAV research and clinical practice for HAV infection in Japan, have been reviewed by the AMED HAV and HEV Study Group.

Outcomes in human T-cell leukemia virus type I carriers after hematopoietic stem cell transplantation for diseases other than adult T cell leukemia/lymphoma: a Japanese national survey

Human T-cell leukemia virus type I (HTLV-1) is a retrovirus known to cause adult T-cell leukemia/lymphoma (ATL). There are few reports on hematopoietic stem cell transplantation (HSCT) for HTLV-1 carriers with diseases other than ATL. A total of 25,839 patients (24,399 adults and 1440 children) with pre-transplant HTLV-1 serostatus information recorded in the Japanese National Survey Database who had undergone their first HSCT were analyzed. We investigated the overall survival (OS), transplant-related mortality (TRM), and disease-related mortality (DRM) after HSCT in relation to HTLV-1 serologic status. Three hundred and forty-eight patients were HTLV-1 antibody carriers. The number of HTLV-1 carriers and noncarriers among adult patients who received allogeneic HSCT (allo-HSCT) or autologous HSCT (auto-HSCT) was 237/15,777 and 95/8920, respectively, and was 16/1424 among pediatric patients who received allo-HSCT. No pediatric HTLV-1 carrier recipients undergoing auto-HSCT were identified. There were no significant differences between HTLV-1 carriers and non-carriers regarding stem cell source, disease risk, or HCT-CI score prior to allo-HSCT. Multivariate analysis of OS (P=0.020) and TRM (P=0.017) in adult patients showed that HTLV-1 positive status was a significant prognostic factor. In children, TRM was significantly higher (P=0.019), but OS was not significantly different. In adult patients who underwent auto-HSCT, HTLV-1 positive status was not a significant prognostic factor. In adult allo-HSCT patients, cytomegalovirus reactivation was significantly more common in HTLV-1 carriers (P=0.001). HTLV-1 antibody positivity was shown to have a poor prognosis in OS and TRM after allo-HSCT in adult patients and in TRM after allo-HSCT in pediatric patients. This work was supported in part by the practical research programs of the Japan Agency for Medical Research and Development (AMED) under grant number 17ck0106342h0001.

Safety, efficacy and pharmacokinetics of palivizumab in off-label neonates, infants, and young children at risk for serious respiratory syncytial virus infection: a multicenter phase II clinical trial

Pediatric patients with certain rare diseases are at increased risk of severe respiratory syncytial virus (RSV) infection. However, the prophylactic use of anti-RSV antibody (palivizumab) in these patients is not indicated at present in Japan. This first-in-the-world multicenter, uncontrolled, open-label, phase II clinical trial was carried out between 28 July 2019 and 24 September 2021at seven medical institutions in Japan to investigate the efficacy, safety, and pharmacokinetics of palivizumab in 23 subjects recruited from among neonates, infants, or children aged 24 months or younger who had any of the following conditions: pulmonary hypoplasia, airway stenosis, congenital esophageal atresia, inherited metabolic disease, or neuromuscular disease. At least four continuous doses of palivizumab were administered intramuscularly at 15mg/kg at intervals of 30 days. Twenty-three enrolled subjects completed the study. No subject required hospitalization for RSV. Adverse events (AE) did not notably differ from the event terms described in the latest interview form. Five severe AEs required unplanned hospitalization, but resolved without RSV infection. Therapeutically effective concentrations of palivizumab were maintained throughout the study period. Palivizumab might be well tolerated and effective in preventing serious respiratory symptoms and hospitalization due to severe RSV infection, indicating the prophylactic use in the pediatric patients included in this study. Japan Agency for Medical Research and Development (AMED), grant numbers 19lk0201097h0001 (to MM), 20lk0201097h0002 (to MM), 21lk0201097h0003 (to MM), and 22lk0201097h0004 (to MM). AMED did not have any role in the execution of this study, analysis and interpretation of the data, or the decision to submit the results.

THE GENETIC SUBTYPES AND THE TUMOR MICROENVIRONMENT SIGNATURES ARE ASSOCIATED WITH DISTINCT OUTCOMES IN PERIPHERAL T‐CELL LYMPHOMA

Background: Emerging evidence suggests the prognostic impact of the tumor microenvironment (TME) in peripheral T-cell lymphomas (PTCLs). To better understand PTCL pathobiology, we performed an integrative multi-omics study to explore the genetic subtypes and the TME signatures of PTCLs, especially nodal T-follicular helper T-cell lymphomas (nTFHLs) and PTCL, not otherwise specified (PTCL-NOS). Method: We performed whole-exome sequencing of 92 nTFHLs and 37 PTCL-NOSs, a total of 129 cases. Ninety-two nTFHLs included 85 angioimmunoblastic T-cell lymphomas (AITLs), four nPTCL with TFH phenotypes, two follicular T-cell lymphomas, and one unclassifiable nTFHL. Genetic drivers were integrated using non-negative matrix factorization consensus clustering (NMF). We estimated the proportion of immune cell fractions from bulk RNA sequencing data (n = 57) using CIBERSORTx and classified the TME signatures using hierarchical clustering. Results: NMF clustering analysis revealed three genetic subtypes within our cohort, except for 10 cases lacking recurrent genetic abnormalities (denoted as cluster 0). C2 was characterized by TP53 mutation, CDKN2A loss, PTEN loss, and chromosomal instability, corresponding to the previously described GATA3-PTCL subtype. C1 and C3 shared TFH-related genomic alterations, but C3 had significantly higher frequencies of chromosome (Chr) 5 gain, Chr 21 gain, RHOA, IDH2, and CD28 mutations. A higher frequency of rash and higher C-reactive protein levels than the other AITLs characterized C3-AITL. Pairwise association analysis of genomic alterations revealed that multiple TET2 mutations, rather than a single TET2 mutation, significantly co-occurred with IDH2 and RHOA mutations. A Bradley-Terry model estimated that the first TET2 and DNMT3A mutations occurred earlier, and the second or later TET2, RHOA, and IDH2 mutations occurred later. Compared to C1, significantly inferior survival was observed in C2 (hazard ratio (HR), 2.52; 95% CI, 1.37–4.63) and C3(HR, 2.14; 95% CI, 1.17–3.89; Figure A). The TME signatures were divided into TME1 (B-cells and follicular helper T-cells), TME2 (activated memory CD4 T-cells, M2 macrophages, and CD8 T-cells), and TME3 (naïve CD4 T-cells and activated mast cells). mTORC1 signaling was enriched in the TME2 signature compared to TME1 and TME3 signatures. The TME2 signature was significantly associated with shorter survival than the TME3 signature (HR: 3.4, 95% CI 1.6–7.5; Figure B). C2 had significantly more TME2 signature than C1 (64.3% vs. 7.7%), while C1 had significantly more TME3 signature than C2 (80.8% vs. 28.6%, p = 0.006; Figure C). Keywords: genomics, epigenomics, and other -omics, microenvironment Encore Abstract—previously submitted to EHA 2023 The research was funded by: Japan Society for the Promotion of Science KAKENHI grants (grant numbers: 19K23879, 22K19451, and 21H02945) and Japan Agency for Medical Research and Development (AMED) grants (20ck0106544 and 21ck0106644)

FIVE‐YEAR RESULTS OF A PHASE 2 STUDY OF CNS‐ORIENTED THERAPY WITH R‐CHOP FOR UNTREATED INTRAVASCULAR LARGE B‐CELL LYMPHOMA: FINAL ANALYSIS OF THE PRIMEUR‐IVL STUDY

Introduction: Intravascular large B-cell lymphoma (IVLBCL) is a rare disease entity of extranodal large B-cell lymphoma characterized by selective growth of lymphoma cells in the lumina of small vessels. The prognosis of IVLBCL is typically poor without timely diagnosis. Based on the promising efficacy of rituximab-containing chemotherapy and a high incidence of secondary central nervous system (CNS) involvement after rituximab-chemotherapy, we conducted the PRIMEUR-IVL study to explore the efficacy of CNS-oriented therapy with rituximab-chemotherapy whose primary analysis demonstrated 2-year progression-free survival (PFS) of 76% and 2-year overall survival (OS) of 92% with a low incidence of secondary CNS involvement of 3%. Patients and Methods: The PRIMEUR-IVL study is a multicenter, single-arm, phase 2 trial at 22 hospitals in Japan for untreated histologically confirmed IVLBCL patients without apparent CNS involvement at diagnosis. The treatment regimen includes three cycles of R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisolone) followed by two cycles of rituximab with high-dose methotrexate and additional three cycles of R-CHOP. Intrathecal chemotherapy including methotrexate, cytarabine, and prednisolone was administered four times during the R-CHOP phase. We present a preplanned final analysis of the PRIMEUR-IVL study including 5-year PFS, OS and secondary CNS involvement. Results: A total of 38 patients were enrolled, of whom 37 patients were eligible. One patient with a history of testicular lymphoma was excluded. As of July 2021, with a median follow-up of 7.1 years (interquartile range 5.6–8.7), 5-year PFS in all eligible patients was 68% (95% confidence interval [CI] 50%–80%) and OS was 78% (95% CI 61%–89%). Median PFS and OS were not reached. No additional secondary CNS involvement was observed after primary analysis with 3% of cumulative incidence. Severe adverse events after the primary analysis were grade 4 neutropenia (n = 1) and grade 4 myelodysplastic syndrome that did not require specific treatment (n = 1). During the observation period after enrollment, there were a total of eight deaths due to following reasons: primary disease (n = 6), sepsis (n = 1), and unknown sudden death (n = 1). Conclusions: In this 5-year, long-term follow-up analysis of the PRIMEUR-IVL study, frontline treatment of CNS-oriented therapy with standard R-CHOP for untreated IVLBCL patients without apparent CNS involvement provided durable response and clinically meaningful results for PFS, OS, and low incidence of secondary CNS involvement with manageable toxicity. Our result provides one of active treatment for untreated IVLBCL patients. The research was funded by: The Practical Research for Innovative Cancer Control, the Japan Agency for Medical Research and Development (AMED), Japan, JP19ck0106511 and the National Cancer Center Research and Development Fund, 23-A-17 and 26-A-4. The research was supported by the Center for Supporting Hematology-Oncology Trials (C-SHOT). Keyword: chemotherapy Conflicts of interests pertinent to the abstract K. Shimada Honoraria: AstraZeneca, Chugai, Eisai, Kyowa Kirin, Symbio M. Yamaguchi Honoraria: AbbVie, Bristol Myers Squibb, Chugai, Janssen, Kyowa Kirin, Meiji Seika, MSD, Nippon Shinyaku, SymBio, Takeda Research funding: AstraZeneca, Chugai Pharma, Genmab, Incyte, Eisai, Takeda, Nippon Shinyaku, Otsuka, Asahi Kasei, Kyowa Kirin, Chugai H. Nagai Honoraria: Kyowa Kirin, Takeda, Chugai, Bristol Myers Squibb, Eisai, Novartis, Janssen, Mundi Pharma, Lilly, AstraZeneca, Ono, Meiji, AbbVie, Nippon Shinyaku, GSK, Sumitomo Pharma, Genmab, CSL Behring Research funding: Kyowa Kirin, Takeda, Chugai, Bristol Myers Squibb, Janssen, AstraZeneca, Ono, AbbVie, Nippon Shinyaku, Daiichi-Sankyo, Zenyaku Kogyo, Solasia J. Takizawa Honoraria: Chugai, Kyowa Kirin N. Fukuhara Honoraria: Chugai, Genmab, AbbVie, Takeda, Eli Lilly, AstraZeneca, Meiji Seika, Ono, Janssen, Bristol-Myers Squibb, Eisai, Kyowa Kirin, Symbio, Novartis Research funding: Chugai, Genmab, AbbVie, Takeda, Eli Lilly, Incyte, Chordia Therapeu D. Ennishi Honoraria: Eisai, Kyowa Kirin, Chugai Research funding: Nippon Shinyaku, Chugai Y. Minami Consultant or advisory role: Takeda, Novartis Honoraria: Bristol-Myers Squibb, Novartis, Pfizer, Daiichi Sankyo, Astellas Research funding: CMIC, Bristol-Myers Squibb, Novartis, Takeda, Chugai, Ono Y. Atsuta Consultant or advisory role: Meiji Seika, JCR Pharmaceuticals, Kyowa Kirin Honoraria: Novartis, AbbVie H. Kiyoi Honoraria: Novartis, Astellas, AbbVie, Chugai Research funding: Zenyaku Kogyo, Nippon Shinyaku, Chugai, Astellas, Kyowa Kirin, Takeda, Sumitomo Pharma, Sanofi, Eisai, Ono, FUJIFILM, Kyowa Kirin, Otsuka, Perseus Proteomics, Daiichi Sankyo, CURED, Bristol-Myers Squibb, AbbVie

CD8 +T-cell memory induced by SARS-CoV-2 mRNA vaccination is maintained by clonal replenishment

Abstract mRNA vaccines against the Spike glycoprotein of severe acute respiratory syndrome type 2 coronavirus (SARS-CoV-2) elicit strong CD8 +T-cell responses. However, in repetitive vaccination, it is unknown whether T-cell clones induced by the first dose respond repeatedly or new clones are mobilized and respond after each dose. Here, we temporally tracked the CD8 +T-cell repertoire in individuals who received three doses of the BNT162b2 mRNA vaccine. CD8 +T-cell clones that expanded after the first, second, or third dose were identified, and their kinetic profiles throughout the vaccination were analyzed. The extent of clonal expansion after the first vaccination was significantly less than that after the second or third vaccination. After the second vaccination, clones that had expanded after the first dose (“first responders”) were less responsive, while a new set of clones (“second responders”) greatly expanded and surpassed the first responders. After the third vaccination, the second responders expanded again, while the first responders did not. Moreover, we observed expansion of another set of clones that had not responded to the previous doses (“third responders”). Single-cell mRNA/protein/TCR analysis after the second vaccination revealed that the first-responder clones exhibited a terminally differentiated phenotype, whereas the second-responder clones exhibited an actively proliferating phenotype. These results suggest that CD8 +T-cell memory induced by repetitive mRNA vaccination is maintained by the mobilization of newly generated clones with proliferative potential. This work was supported by Japan Society for the Promotion of Science (JSPS) under Grant Number 17929397 and 20281832, and Japan Agency for Medical Research and Development (AMED) under Grant Number JP22fk0310509, JP22ama221306, and P21gm6210025.

A combination of TLR9 and STING agonists induces potent neopeptide-specific T cell immunity and improves ICB efficacy in tumor models

Abstract Immunotherapies such as immune checkpoint blockade (ICB) therapies have been developed in the last years as a promising strategy for the treatment of cancer, however there remains a need to improve their efficacy. The most critical factor that affects the efficacy of ICB therapies, is the frequency of tumor mutations, the associated neoantigens generated and the T-cell response against them. Therefore, it is expected that neoantigen vaccinations would improve ICB therapy efficacy by boosting the neoantigen-specific T cell response. The aim of this study is to develop an effective vaccine using a combination of TLR9 and STING agonists together with synthetic long peptides to induce a potent neoantigen-specific cytotoxic CD8 +T cell response. We first analyzed the efficacy of the vaccine formulation containing 20-mer OVA peptide and found that mice immunized with OVA peptide together with the adjuvant combination induced potent antigen-specific T cell responses and are capable of controlling tumor growth and improving survival in B16-F10-OVA tumor bearing mice. Moreover, we found that the vaccine together with OVA peptide is able to synergize with anti-PD-1 treatment in controlling tumor growth. Finally, we examined the immunogenicity of the vaccine formulation with neopeptides identified from two different tumor models. The vaccine formulation induced potent neoantigen-specific T cell responses in vivo. Our findings demonstrate that the vaccine formulation using the combination of TLR9 and STING agonists induces potent T cell immunity against synthetic long peptides and can serve as a promising immunogenic neoantigen vaccine platform. Supported by the Department of Immunology of the Erasmus MC the Dutch Cancer Society (KWF grant 12837) and in part by a grant from International Joint Usage/Research Center, the Institute of Medical Science, the University of Tokyo, and Japan Agency for Medical Research and Development (AMED).

MP69-02 CLINICAL IMPACT OF CANCER PREDISPOSING GENES ON UPPER URINARY TRACT UROTHELIAL CARCINOMA: A LARGE SCALE GENOME ANALYSIS IN JAPAN

You have accessJournal of UrologyCME1 Apr 2023MP69-02 CLINICAL IMPACT OF CANCER PREDISPOSING GENES ON UPPER URINARY TRACT UROTHELIAL CARCINOMA: A LARGE SCALE GENOME ANALYSIS IN JAPAN Yuya Sekine, Yusuke Iwasaki, Mikiko Endo, Takeshi Sano, Shusuke Akamatsu, Takashi Kobayashi, Hidewaki Nakagawa, Kazuyuki Numakura, Shintaro Narita, Yukihide Momozawa, and Tomonori Habuchi Yuya SekineYuya Sekine More articles by this author , Yusuke IwasakiYusuke Iwasaki More articles by this author , Mikiko EndoMikiko Endo More articles by this author , Takeshi SanoTakeshi Sano More articles by this author , Shusuke AkamatsuShusuke Akamatsu More articles by this author , Takashi KobayashiTakashi Kobayashi More articles by this author , Hidewaki NakagawaHidewaki Nakagawa More articles by this author , Kazuyuki NumakuraKazuyuki Numakura More articles by this author , Shintaro NaritaShintaro Narita More articles by this author , Yukihide MomozawaYukihide Momozawa More articles by this author , and Tomonori HabuchiTomonori Habuchi More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003332.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Quite a few numbers of upper urinary tract urothelial carcinoma (UTUC) would be caused by germline pathogenic variants. Although mismatch repair (MMR) genes are known for pathogenicity for UTUC, the impact of other cancer-predisposing genes, such as BRCA1/2, on UTUC has never been thoroughly investigated despite the fact that panel sequencing is a standard method for advanced cancers. We conducted a case-control genetic analysis to clarify the association between these possible genes and UTUC in Japanese. METHODS: We enrolled 210 patients with UTUC and 38,300 general subjects from Biobank Japan as controls. Twenty-seven genes were sequenced, including cancer-predisposing genes such as MMRs and BRCA1/2. Pathogenic variants were defined as follows; loss-of-function variants or variants registered as pathogenic in ClinVar. RESULTS: After quality control, 209 patients with UTUC and 37,727 controls were analyzed. In the 27 analyzed genes, 4,769 germline variants, including 286 pathogenic variants, were confirmed. The pathogenic variants in MMR genes were detected in 1.44% of patients with UTUC and significantly associated with conceiving the UTUC (p=1.30×10-3, OR 15.24). In the other genes, BRCA1 and BRCA2 had only one patient with pathogenic variants, and no significant association was verified. CONCLUSIONS: The significant association was observed between MMR genes and the prevalence of UTUC in Japanese. Other cancer-predisposing genes were not associated with UTUC in our study. It needs more patients to know whether these genes affect the risk of UTUC. Source of Funding: Japan Agency for Medical Research and Development (AMED) under Grant No. JP19kk0305010 © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e963 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Yuya Sekine More articles by this author Yusuke Iwasaki More articles by this author Mikiko Endo More articles by this author Takeshi Sano More articles by this author Shusuke Akamatsu More articles by this author Takashi Kobayashi More articles by this author Hidewaki Nakagawa More articles by this author Kazuyuki Numakura More articles by this author Shintaro Narita More articles by this author Yukihide Momozawa More articles by this author Tomonori Habuchi More articles by this author Expand All Advertisement PDF downloadLoading ...

MP01-10 A NOVEL AND FEASIBLE METHOD FOR QUANTIFYING TESTICULAR SPERAMATOGENESIS BY USING CREATINE CHEMICAL EXCHANGE SATURATION TRANSFER IMAGING

You have accessJournal of UrologyCME1 Apr 2023MP01-10 A NOVEL AND FEASIBLE METHOD FOR QUANTIFYING TESTICULAR SPERAMATOGENESIS BY USING CREATINE CHEMICAL EXCHANGE SATURATION TRANSFER IMAGING Sohei Kuribayashi, Shinichiro Fukuhara, Go Tsujimura, Takahiro Imanaka, Koichi Okada, Norichika Ueda, Kentaro Takezawa, Shigeyoshi Saito, Yusuke Takahashi, Hidetaka Kioka, and Norio Nonomura Sohei KuribayashiSohei Kuribayashi More articles by this author , Shinichiro FukuharaShinichiro Fukuhara More articles by this author , Go TsujimuraGo Tsujimura More articles by this author , Takahiro ImanakaTakahiro Imanaka More articles by this author , Koichi OkadaKoichi Okada More articles by this author , Norichika UedaNorichika Ueda More articles by this author , Kentaro TakezawaKentaro Takezawa More articles by this author , Shigeyoshi SaitoShigeyoshi Saito More articles by this author , Yusuke TakahashiYusuke Takahashi More articles by this author , Hidetaka KiokaHidetaka Kioka More articles by this author , and Norio NonomuraNorio Nonomura More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003212.10AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The sperm retrieval rate in non-obstructive azoospermia (NOA) patients is still insufficient. To overcome this problem, an accurate noninvasive method of evaluating testicular spermatogenesis is needed. Chemical exchange saturation transfer (CEST) imaging is a new magnetic resonance imaging (MRI) technique that can image the distribution of trace substances in vivo. We focused on the potential role of creatine (Cr) in testes and hypothesized that Cr-CEST could indicate intratesticular spermatogenesis noninvasively. METHODS: We investigated the Cr-CEST signal in the testes of the testicular ischemia model and the partial testicular irradiation model. In the testicular ischemia model, we clamped the right spermatic cord from 6–42 hours. To evaluate spatial resolution, we performed Cr-CEST for a partial testicular irradiation model in which the lower half of the testes was irradiated with a single 6 Gy dose. RESULTS: In the testicular ischemia models, eight testicular ischemia models were serially imaged at two or three-time points and a total of 26 images of ischemic testis were obtained. The signal intensity of ischemic testis was significantly lower than that of controls (4.0±2.0 vs. 8.9±0.6, p<0.01, Figure 1). Correlation analysis revealed a strong linear correlation between the signal intensity decline and the duration of ischemia (r=0.96, p<0.01, Figure 1). In the partial irradiation model, the signal intensity in the irradiated area was significantly lower than that in the non-irradiated area (5.3± 1.4 vs 7.6±0.3, p<0.05, Figure 1). CONCLUSIONS: Cr-CEST noninvasively evaluate intratesticular spermatogenesis both qualitatively and spatially from this study. Cr-CEST would be used as feasible preoperative imaging for evaluating intratesticular spermatogenesis and a navigation system during microscopic intratesticular sperm extraction. Source of Funding: This research was supported by the Japan Agency for Medical Research and Development (AMED) (grant number: J210705035) and Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS) (grant number: A21H030670) © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e5 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Sohei Kuribayashi More articles by this author Shinichiro Fukuhara More articles by this author Go Tsujimura More articles by this author Takahiro Imanaka More articles by this author Koichi Okada More articles by this author Norichika Ueda More articles by this author Kentaro Takezawa More articles by this author Shigeyoshi Saito More articles by this author Yusuke Takahashi More articles by this author Hidetaka Kioka More articles by this author Norio Nonomura More articles by this author Expand All Advertisement PDF downloadLoading ...

MP04-13 IMMUNOPHENOTYPE ANALYSIS OF TUMOR-INFILTRATING IMMUNE CELLS TO ELUCIDATE THE MECHANISM OF ANTITUMOR EFFECT OF IL-7 AND CCL19 PRODUCING CAR-T CELLS AGAINST SOLID CANCER

You have accessJournal of UrologyCME1 Apr 2023MP04-13 IMMUNOPHENOTYPE ANALYSIS OF TUMOR-INFILTRATING IMMUNE CELLS TO ELUCIDATE THE MECHANISM OF ANTITUMOR EFFECT OF IL-7 AND CCL19 PRODUCING CAR-T CELLS AGAINST SOLID CANCER Shunsuke Goto, Koji Tamada, and Masatoshi Eto Shunsuke GotoShunsuke Goto More articles by this author , Koji TamadaKoji Tamada More articles by this author , and Masatoshi EtoMasatoshi Eto More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003215.13AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Chimeric antigen receptor (CAR)-T cell therapy against solid cancer is challenging. We reported that mouse CAR-T cells modified to produce interleukin- (IL)-7 and CCL19 (7×19 CAR-T) enhance the therapeutic efficacy against solid tumors by recruiting not only administered CAR-T cells but also endogenous T cells and DCs into tumors and inducing long-term antitumor memory responses. Furthermore, we reported that human 7×19 CAR-T cells show enhanced antitumor efficacy against patient-derived xenograft (PDX) tumor expressing mesothelin, for which 10% of renal cell carcinoma are positive. In this research, we explore the mechanism of antitumor activity of 7×19 CAR-T cells and object to prevail the difference between conventional CAR-T cells (Conv.) and 7×19, especially focusing on the immunophenotype of tumor-infiltrating lymphocytes (TILs). METHODS: We produced a novel anti-human mesothelin 7×19 CAR-T cells, then assessed the antitumor activity against solid tumor in vivo using a malignant mesothelioma orthotopic tumor and a pancreatic cancer PDX models. Tumor was resected after the administration of Conv. or 7×19 CAR-T cells and analyzed TILs by flowcytometry focusing on the number, exhaustion marker, phenotype and cytokine producing ability. RESULTS: 7×19 CAR-T cells showed superior antitumor efficacy compared to Conv. CAR-T cells in the orthotopic and PDX tumor models. In the PDX treatment model, TIL analysis revealed that not only CAR-T cells but also non-CAR-T cells were infiltrated effectively into tumors by 7×19 treatment compared to Conv. Exhaustion marker expressions such as PD-1 and TIGIT were significantly downregulated in 7×19 than Conv. The examination of the ability of TILs to express cytokines and effector molecules showed that significant increases of TNF-α and Granzyme B, as well as a trend of increased IFN-γ in 7×19 CAR-T cells. The analysis of splenocyte after the treatment revealed that 7×19 CAR-T cells maintained their effector memory phenotype. CONCLUSIONS: It was found that 7×19 CAR-T cells showed enhanced antitumor potential by the maintenance of their effector functions. These data support the foresight of this therapy as a treatment option for patients with mesothelin-positive renal cell carcinoma. Source of Funding: Practical Research for Innovative Cancer Control, and Project for Cancer Research and Therapeutic Evolution (P-CREATE) 16770206 by Japan Agency for Medical Research and Development (AMED) © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e38 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Shunsuke Goto More articles by this author Koji Tamada More articles by this author Masatoshi Eto More articles by this author Expand All Advertisement PDF downloadLoading ...

Development of Microphysiological Systems (MPSs) Based on Microfluidic Technology for Drug Discovery in Japan

Microphysiological systems (MPSs) based on microfluidic devices are attracting attention as an alternative cell assay platform to animal experiments in drug discovery. When we use microfluidic devices for cell culture, it is possible to experiment with various culture conditions that are difficult with conventional cell culture methods, such as fabrication of microstructures for cell placement, temporal and spatial control of liquid factors and adhesive conditions, and physical stimulation by flow and expansion/contraction. MPSs, which use microfluidic technology to construct the structure and function of physiological biological tissues and organs, are being commercialized and put to practical use worldwide with the entry of venture companies and pharmaceutical companies. Although research on the practical application of MPS in Japan has lagged far behind the efforts of Western countries, the Japan Agency for Medical Research and Development (AMED) launched the MPS Development and Research Project in FY2017 and established a system for MPS commercialization through industry-government-academia collaboration. The project is characterized by the formation of a consortium involving many researchers not only from academia but also from manufacturing and pharmaceutical companies with the aim of commercializing MPS devices. By FY2021, the final year of this project, several MPSs were successfully positioned in various stages of commercialization. This paper introduces two MPSs that the author was involved in commercializing in collaboration with domestic companies within the project.

A Strategic Translational Research System for Drug Discovery in Tottori University.

The probability of successful drug discovery is declining, and research and development costs are increasing. To solve these problems, pharmaceutical companies tend to in-license seeds from venture companies and academia. Therefore, academia's role in drug discovery is extremely important. Tottori University started a "Next-Generation Research Support Project (Strategic Research Support Project)" in 2020, developing a translational research system to promote drug discovery. In this project, we established a research and development infrastructure, such as seed registration, construction of drug research and development support, and research fund allocation. The registered seed were converted into project, and the project implemented this research and development system, and evaluated and verified its results. Twenty-two seeds were converted into projects and portfolios were constructed. Research funds were allocated to eight prioritized projects. Each project raised the research and development stages. From the overall portfolio, one project with the Japan Agency for Medical Research and Development (AMED) Drug Discovery Booster Project, and three projects with Seeds A of the AMED Translational Research Strategic Promotion Program were adopted. Additionally, a new low-molecular weight chaperone drug against GM1-gangliosidosis was out-licensed to an overseas pharmaceutical company. The strength of this system was the strategic allocation of research funds and the accompanying support that leveraged internal and external resources with the PM and researchers at its core. This system achieved certain results in promoting drug discovery; however, resource optimization of specialized personnel needs to be strengthened in the future. In this report, we summarized the efforts of translational research in Japan and around the world. In addition, the translational research efforts of Japanese academia and Tottori University were compared and the current status was summarized.

STNM01, the RNA oligonucleotide targeting carbohydrate sulfotransferase 15, as second-line therapy for chemotherapy-refractory patients with unresectable pancreatic cancer: An open label, phase I/IIa trial.

The impact of stroma-targeting therapy on tumor immune suppression is largely unexplored. An RNA oligonucleotide, STNM01, has been shown to repress carbohydrate sulfotransferase 15 (CHST15) responsible for tumor proteoglycan synthesis and matrix remodeling. This phase I/IIa study aimed to evaluate the safety and efficacy of STNM01 in patients with unresectable pancreatic ductal adenocarcinoma (PDAC). This was an open-label, dose-escalation study of STNM01 as second-line therapy in gemcitabine plus nab-paclitaxel-refractory PDAC. A cycle comprised three 2-weekly endoscopic ultrasound-guided locoregional injections of STNM01 at doses of 250, 1,000, 2,500, or 10,000nM in combination with S-1 (80-120mg twice a day for 14 days every 3 weeks). The primary outcome was the incidence of dose-liming toxicity (DLT). The secondary outcomes included overall survival (OS), tumor response, changes in tumor microenvironment on immunohistopathology, and safety (jRCT2031190055). A total of 22 patients were enrolled, and 3 cycles were repeated at maximum; no DLT was observed. The median OS was 7.8 months. The disease control rate was 77.3%; 1 patient showed complete disappearance of visible lesions in the pancreas and tumor-draining lymph nodes. Higher tumoral CHST15 expression was associated with poor CD3+ and CD8+ T cell infiltration at baseline. STNM01 led to a significant reduction in CHST15, and increased tumor-infiltrating CD3+ and CD8+ T cells in combination with S-1 at the end of cycle 1. Higher fold increase in CD3+ T cells correlated with longer OS. There were 8 grade 3 adverse events. Locoregional injection of STNM01 was well tolerated in patients with unresectable PDAC as combined second-line therapy. It prolonged survival by enhancing T cell infiltration in tumor microenvironment. The present study was supported by the Japan Agency for Medical Research and Development (AMED).

Liver autophagy-induced valine and leucine in plasma reflect the metabolic effect of sodium glucose co-transporter 2 inhibitor dapagliflozin.

Sodium glucose co-transporter 2 (SGLT2) inhibitors are anti-diabetic drugs for type 2 diabetes that lower blood glucose levels and body weight. It is of special interest that SGLT2 inhibitors also improve liver metabolism and fatty liver. Liver is an important organ in regulation of energy metabolism, but the metabolic action of SGLT inhibitors in liver remains unclear. We investigated the factors associated with the beneficial effects of dapagliflozin, a SGLT2 inhibitor, in the liver after confirming its glucose-lowering and weight loss effects using an obesity and diabetes mouse model. We also performed clinical study of patients with type 2 diabetes to explore candidate biomarkers that reflect the beneficial action of dapagliflozin in the liver. In animal study, dapagliflozin induced autophagy in the liver (LC3-II to LC3-I expression ratio: P<0·05 vs. control), and valine and leucine levels were increased in plasma (P<0·01 vs. control) as well as in liver (P<0·05 vs. control). Thus, increased plasma valine and leucine levels are potential biomarkers for improved liver metabolism. Clinical study found that valine and leucine levels were markedly higher in patients treated with dapagliflozin (valine: P<0·05 vs. control, leucine: P<0·01 vs. control) than those not treated after one week intervention. Dapagliflozin improves liver metabolism via hepatic autophagy, and plasma valine and leucine levels may reflect its metabolic effect. AstraZeneca K.K., Ono Pharmaceutical Co., Ltd., Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan Society for the Promotion of Science (JSPS), Japan Agency for Medical Research and Development (AMED), Novo Nordisk Pharma Ltd., and Japan Foundation for Applied Enzymology, and MSD Life Science Foundation International.

A clinical-grade HLA haplobank of human induced pluripotent stem cells matching approximately 40% of the Japanese population

Human induced pluripotent stem cells (iPSCs) are expected to be useful for regenerative medicine for many diseases. Many researchers have focused on and enabled the generation of differentiated cells or tissue-like structures, including organoids, which help to ameliorate target diseases. To promote such cell therapies, we established a clinically applicable iPSC haplobank matching as many people as possible in Japan. Through cooperation with several organizations, we recruited donors whose human leukocyte antigens (HLAs) involved in immunorejection were homozygous. The peripheral or umbilical cord blood collected from the donors was used for iPSC production by electroporation of episomal vectors. These iPSC lines were then subjected to testing, including genome analyses and sterility, to maximize safety. We constructed a clinical-grade haplobank of 27 iPSC lines from 7 donors according to good manufacturing practice regulations. However, reasons to avoid using iPSC lines include the presence of residual episomal vectors or genetic mutations in cancer-related genes. This haplobank provides HLA-matched iPSC lines for approximately 40% of the Japanese population. Since the haplobank's release in 2015, these iPSC lines have been used in more than 10 clinical trials. The establishment of this haplobank is an important step toward the clinical application of iPSCs in cell therapies. This study was supported by a research center network for therealization of regenerative medicine of the Japan Agency for Medical Research and Development (AMED) under grant number JP20bm0104001h0108.

Points to Consider for Continuous Manufacturing of Biotechnological Drug Products

Continuous Manufacturing is a manufacturing method in which raw materials enter the manufacturing process continuously, and products are discharged continuously throughout the duration of the process including multiple unit operations. Continuous Manufacturing includes various options: Those with all stages of processing from charging raw materials to discharging final products that are continuous, and those with only certain parts of the manufacturing process that are continuous. It can be introduced in an appropriate range in consideration of the characteristics of products, manufacturing processes, robustness of processes, etc. Continuous Manufacturing is expected to have the following advantages: A wide range of manufacturing scales are available by adjusting the continuous operation time of the process; it is possible to use the same manufacturing equipment from the development stage to commercial production, eliminating the need to change to large scale manufacturing equipment; and it is possible to reduce the equipment installation area (footprint). While Continuous Manufacturing can be a major manufacturing technique of the pharmaceutical industry in the future, official documents such as guidelines for Continuous Manufacturing are limited. In particular, no official documents such as guidelines specialized in manufacturing of biotechnological drug products have been issued yet. In Japan Agency for Medical Research and Development (AMED) research project, we summarized the key points to consider for investigation of the control strategy for Continuous Manufacturing of biotechnological products as a document. In this paper, we introduce the document.

Favipiravir, camostat, and ciclesonide combination therapy in patients with moderate COVID-19 pneumonia with/without oxygen therapy: An open-label, single-center phase 3 randomized clinical trial.

SummaryBackgroundThe effectiveness of combination therapy for COVID-19 pneumonia remains unclear. We evaluated favipiravir, camostat, and ciclesonide combination therapy in patients with moderate COVID-19 pneumonia.MethodsIn this open-label phase 3 study, hospitalized adults who were positive for SARS-CoV-2 and had COVID-19 pneumonia were enrolled prior to official vaccination drive in Japan. Participants were randomly assigned to favipiravir monotherapy or favipiravir + camostat + ciclesonide combination therapy. The primary outcome was the length of hospitalization due to COVID-19 infection after study treatment. The hospitalization period was calculated from the time of admission to the time of patient discharge using the clinical management guide of COVID-19 for front-line healthcare workers developed by the Japanese Ministry of Health, Labour, and Welfare (Version 3). Cases were registered between November 11, 2020, and May 31, 2021. Japan Registry of Clinical Trials registration: jRCTs031200196.FindingsOf 121 enrolled patients, 56 received monotherapy and 61 received combination therapy. Baseline characteristics were balanced between the groups. The median time of hospitalization was 10 days for the combination and 11 days for the monotherapy group. The median time to discharge was statistically significantly lower in the combination therapy vs monotherapy group (HR, 1·67 (95% CI 1·03–2·7; P = 0·035). The hospital discharge rate was statistically significantly higher in the combination therapy vs monotherapy group in patients with less severe COVID-19 infections and those who were ≤60 years. There were no significant differences in clinical findings between the groups at 4, 8, 11, 15, and 29 days. Adverse events were comparable between the groups. There were two deaths, with one in each group.InterpretationCombination oral favipiravir, camostat and, ciclesonide therapy could decrease the length of hospitalization stays without safety concerns in patients with moderate COVID-19 pneumonia. However, lack of hard clinical primary outcome is one of the major limitations of the study.FundingThis research was supported by Japan Agency for Medical Research and Development (AMED) under Grant Number 20fk0108261h0001.

MP12-03 IL-7 AND CCL19 PRODUCING CAR-T CELLS PREVENT ANTIGEN-LOSS TUMOR RELAPSE AND ENHANCE ANTITUMOR EFFICACY AGAINST SOLID CANCER

You have accessJournal of UrologyCME1 May 2022MP12-03 IL-7 AND CCL19 PRODUCING CAR-T CELLS PREVENT ANTIGEN-LOSS TUMOR RELAPSE AND ENHANCE ANTITUMOR EFFICACY AGAINST SOLID CANCER Shunsuke Goto, Koji Tamada, and Masatoshi Eto Shunsuke GotoShunsuke Goto More articles by this author , Koji TamadaKoji Tamada More articles by this author , and Masatoshi EtoMasatoshi Eto More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002534.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: While chimeric antigen receptor (CAR)-T cell brought a breakthrough to the treatment for hematological cancer, most CAR-T cell therapies failed to treat solid cancers. We recently reported that mouse CAR-T cells modified to produce interleukin- (IL)-7 and CCL19 (7×19 CAR-T cells) enhance the therapeutic efficacy against solid tumors by recruiting not only administered CAR-T cells but also endogenous T cells and DCs into tumors. Further, we reported that human 7×19 CAR-T cells show enhanced antitumor efficacy against PDX tumor expressing mesothelin, for which 10% of renal cell carcinoma are positive. In this research, we focus on the mechanism of antitumor activity of 7×19 CAR-T cells and object to prevail the difference between conventional CAR-T cells (Conv.) and 7×19. Another hurdle of CAR-T cell therapy is adverse event arising from unspecific reaction to normal tissues and organs. In this regard, we investigated the antigen-specific activity of 7×19 CAR-T cells by conducting a tumor rechallenge experiment. METHODS: We produced a novel anti-human mesothelin 7×19 CAR-T cells, then assessed the antitumor activity against solid tumor in vivo using a malignant mesothelioma orthotopic tumor model. Tumor was resected after the administration of CAR-T cells and surface expression levels of mesothelin were measured by flowcytometry. Tumor rechallenge experiment was done using PDX tumor model. RESULTS: 7×19 CAR-T cells rejected the orthotopic tumor, while Conv. CAR-T cells allowed the tumor relapse. The surface expression of mesothelin on those relapsing tumors were declined from original tumors, which suggested that 7×19 CAR-T cells prevented the relapse of antigen-loss tumors. In the rechallenge trial, mesothelin-positive tumor progression was specifically suppressed in mice which had rejected mesothelin positive PDX tumors by the treatment with 7×19 CAR-T cells. CONCLUSIONS: It was found that the enhanced antitumor potential of human anti-mesothelin 7×19 CAR-T cells are antigen-specific and show a long-term memory effect. Further, 7×19 CAR-T cells can solve one of the important clinical problems, antigen-loss tumor relapse. These data support that this therapy will be a new treatment option for patients with mesothelin-positive renal cell carcinoma. Source of Funding: This study was supported by Practical Research for Innovative Cancer Control, and Project for Cancer Research and Therapeutic Evolution (P-CREATE) 16770206 by Japan Agency for Medical Research and Development (AMED), and Japan Society for the Promotion of Science Grant-in-Aid for Scientific Research (C) Grant Number 19K07625 © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e166 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Shunsuke Goto More articles by this author Koji Tamada More articles by this author Masatoshi Eto More articles by this author Expand All Advertisement PDF DownloadLoading ...