Discovery of ASP6918, a KRAS G12C inhibitor: Synthesis and structure-activity relationships of 1-{2,7-diazaspiro[3.5]non-2-yl}prop-2-en-1-one derivatives as covalent inhibitors with good potency and oral activity for the treatment of solid tumors.

Although KRAS protein had been classified as an undruggable target, inhibitors of KRAS G12C mutant protein were recently reported to show clinical efficacy in solid tumors. In our previous report, we identified 1-{2,7-diazaspiro[3.5]non-2-yl}prop-2-en-1-one derivative (1) as a KRAS G12C inhibitor that covalently binds to Cys12 of KRAS G12C protein. Compound 1 exhibited potent cellular pERK inhibition and cell growth inhibition against a KRAS G12C mutation-positive cell line and showed an antitumor effect on subcutaneous administration in an NCI-H1373 (KRAS G12C mutation-positive cell line) xenograft mouse model in a dose-dependent manner. In this report, we further optimized the substituents on the quinazoline scaffold based on the structure-based drug design from the co-crystal structure analysis of compound 1 and KRAS G12C to enhance in vitro activity. As a result, ASP6918 was found to exhibit extremely potent in vitro activity and induce dose-dependent tumor regression in an NCI-H1373 xenograft mouse model after oral administration.

Differences in serum selenoprotein P profile between C57BL/6 and BALB/c mice fed high-fat diet.

BACKGROUND: C57BL/6 mice generally show hyperglycaemia and insulin resistance when fed a high-fat diet (HFD) compared to those of BALB/c mice. However, whether these strains also show different expression profiles of selenoprotein P, a diabetes-related hepatokine, after HFD feeding is unclear. We investigated the effects of HFD on body weight, glucose metabolism, and plasma selenoprotein P levels in C57BL/6 and BALB/c mice. METHODS: Male C57BL/6 and BALB/c mice aged seven weeks were divided into normal diet (ND) and HFD groups. Fasting body weights and blood sugar levels were measured weekly. Blood specimens were collected after 16 h of fasting (in weeks 7, 9, and 11) and after 24 h of subsequent refeeding (in weeks 9 and 11) to analyse plasma selenoprotein P and insulin levels. RESULTS: The mean body weight of the HFD group was consistently higher than that of the ND group for both strains. However, a significant elevation in fasting plasma glucose levels from the early stage was observed only in the HFD group of C57BL/6 mice. In BALB/c mice, a difference in fasting glucose levels between the HFD and ND groups was observed after nine weeks. After seven, nine, and eleven weeks, the fasting plasma insulin levels were higher in the HFD group than in the ND group for both strains. During this period, plasma selenoprotein P levels in the HFD group were significantly higher than those in the ND group of C57BL/6 mice. However, BALB/c mice did not show a significant difference in plasma levels of selenoprotein P between the ND and HFD groups. After refeeding, the plasma insulin and selenoprotein P levels increased compared to those observed during fasting in the ND group for both strains. Elevation of insulin levels, but not of selenoprotein P levels, after refeeding was noticed in the HFD group for both strains. Plasma selenoprotein P level after refeeding was significantly lower than that during fasting in the HFD group of C57BL/6 mice. CONCLUSION: Unlike C57BL/6 mice, BALB/c mice did not show elevated fasting plasma selenoprotein P levels despite HFD feeding. Additionally, the pattern of selenoprotein P levels in the plasma after refeeding differed between C57BL/6 and BALB/c mice. These differences in selenoprotein P expression among strains may be related to different susceptibilities of individuals to diabetes.

Public Health Centers' Training Session Programs to Develop Programs on Infection Control Practices for Multidrug-Resistant Organisms in Hospitals in Kawaguchi City, Japan.

Introduction The Kawaguchi City Public Health Center (PHC) conducted training sessions focusing on infection control practices on multidrug-resistant organisms (MDROs) for 19 hospitals and eight affiliated clinics (AFs) with beds in June 2022. Issues with infection control programs were identified via a survey implemented following the training sessions. These included providing feedback on infection control policies for MDROs, hand hygiene compliance programs (HHCPs), environmental cleaning (EC), and training sessions programs that hospitals or AFs with beds (hospitals) intended to implement in the future or develop (to be developed). We planned to examine whether the PHC training sessions programs have an effect on the development of hospital infection control programs designed to address these issues. The purpose of this study is to clarify the training session program provided by the Kawaguchi City PHC, which was effective in developing hospital infection control programs based on the results of the survey conducted after the training session. Methods In June 2023, a second training session that offered information on infection control practices was completed for 30 hospitals. This was followed by sending a questionnaire. We examined infection control programs to be developed and analyzed associations with the first learned information by training session (the first learned information). Results Twenty-four hospitals responded to the survey with a response rate of 80.0%. Half the respondents (12, 50.0%) had prepared for the infection control policy on carbapenem-resistant Enterobacteriaceae (CRE), 11 hospitals (45.8%) had provided feedback on HHC, and four (16.7%) planned to conduct feedback on HHC. HHCPs were planned to be developed by 19 hospitals (79.2%), EC by five hospitals (20.8%), training session by 12 hospitals (50.0%), and screening of MDROs upon hospital admission (AS) by nine hospitals (37.5%). The first learned information, "the prevention of healthcare-associated infections and cost savings by implementing cleaning bundles (the effects of cleaning bundles)," was identified by 10 hospitals (41.7%), and "specific programs on providing feedback effective for developing hand hygiene compliance (specific feedback)" was learned by eight hospitals (33.3%). The first learned information regarding specific feedback was significantly associated with HHCPs to be developed (p = 0.044). The first learned information on the effects of cleaning bundles was significantly associated with HHCPs and HHC feedback to be developed (p = 0.023, 0.034). The training session programs were not significantly connected to EC, training session, or AS to be developed. Conclusions Infection control programs to be developed were linked to the provision of information on numerical effects by implementing specific feedback and cleaning bundles. We suggest that the PHC should develop infection control programs for the hospitals and provide training sessions, including numerical effects.

Smoking cessation behavior in patients with a diagnosis of a non-communicable disease: The impact of perceived disease severity of and susceptibility to the disease.

INTRODUCTION: The Health Belief Model comprises two constructs influencing changed behaviors impacting on health, namely perceived severity and susceptibility. The aim of this study was to identify the impact of the combination of, or interactions between, these two constructs on quitting smoking in smokers with a diagnosis of a non-communicable disease (NCD). METHODS: From the large insurance claims database maintained by JMDC database (JMDC, Tokyo), we extracted data on 13284 participants who smoked. All participants were stratified according to their NCD diagnosis based on perceived severity and susceptibility as follows: Category I (high severity and high susceptibility) - acute myocardial infarction, and lung cancer; Category II (high severity and low susceptibility) - colorectal cancer, and stomach cancer; Category III (low severity and high susceptibility) - asthma, and transient ischemic attack; Category IV (low severity and low susceptibility) - appendicitis, and glaucoma. We performed multi-variable logistic regression analysis and calculated the proportion of those who were smoking at the first health check-up after the diagnosis and every three years thereafter. RESULTS: Using glaucoma as the reference, the adjusted odds ratios for smoking cessation were 14.2 (95% CI: 11.4-17.8) to 14.8 (95% CI: 12.5-17.4) in Category I; 4.5 (95% CI: 3.8-5.4) to 6.6 (95% CI: 5.4-8.0) in Category II; and 1.9 (95% CI: 1.7-2.1) to 2.8 (95% CI: 2.2-3.7) in Category III. In Categories I and II, the proportion of smokers rapidly decreased after diagnosis and mostly remained low thereafter. Smoking cessation rates for Categories I and II were not associated with readiness to improve lifestyles prior to NCD diagnosis. CONCLUSIONS: Our study confirms the significant impact of perceived severity of and susceptibility to the diagnosed disease on smoking cessation. The multiplicative effect of these two constructs at NCD diagnosis represents a 'teachable moment', a window of opportunity, for encouraging successful long-term smoking cessation.

[Report on COVID-19 infection prevention and control training sessions for care homes provided by Kawaguchi city and comparison with those provided by the local governments in Japan].

Objectives　We compared COVID-19 prevention and control information provided to care homes (CHs) by the Kawaguchi City public health center (PHC), which utilizes our precedent advice on nfection, with the information from several local governments (LGs) in Japan. This study aimed to highlight the role of LG-associated doctors in providing information to CHs, utilizing their precedent advice on infection control in CHs and medical facilities. This study analyzed the sector and type of information the LGs should provide to CHs to prevent and control COVID-19.Methods　We compared training sessions on COVID-19 prevention and control information provided to CHs by the Kawaguchi City PHC with training sessions offered by several other LGs in Japan that are available on their websites.Results　The Kawaguchi City PHC has been providing COVID-19 information to CHs when needed, including prevention and control information, through their doctors, utilizing our precedent advice on infection control, management of health conditions of staff and residents, and early detection of COVID-19. In contrast, 68 LGs announced that they have provided training sessions to CHs for the prevention and control of COVID-19 through their official homepages from March to September 2022. These training sessions involved information dissemination by infection control specialist nurses (42.6%), clinic or hospital doctors (32.4%), infection control specialist doctors (11.8%), and staff from LG headquarters, PHC, or LG-associated doctors (51.5%). Among the 68 LGs, 41 provided information that included hand hygiene (95.1%), personal protective equipment (92.7%), proper ventilation (51.2%), and management of staff (90.2%) and resident (58.5%) health conditions. Furthermore, Kawaguchi City PHC and several LGs provided information for the early detection of COVID-19.Conclusion　We suggest that LGs provide COVID-19 training sessions conducted by LG doctors that include management of staff and resident health conditions, provision of early detection information, and utilization of precedent advice on infection in CHs and medical facilities.

Targeted Infection Control Practices in Japanese Hospitals for Multidrug-Resistant Organisms: Guidance From the Public Health Center.

Introduction The study conducted by the Kawaguchi City Public Health Center (PHC) in 2023 on hospital infection control (IC) programs revealed that hospitals can improve their IC programs if the PHC provides training sessions (TSs) that have numerical effects. In this study, we expected that we could help hospitals develop their IC practices by providing targeted guidance. This study aimed to clarify targeted guidance on IC practices and TS programs to develop hospitals'hospitals' IC programs on multidrug-resistant organisms (MDROs) by examining hospitals'hospitals' IC programs in reference to the study conducted in 2023 and other case reports. Methods In June 2022, the Kawaguchi City PHC conducted TSs for 19 hospitals and eight affiliated (AFs) clinics with beds, providing guidelines and practices on infection control (IC) for MDROs. After the TSs, we sent a questionnaire to these hospitals and affiliated clinics. The questionnaire inquired about current and planned IC policies, hand hygiene compliance programs (HHCPs), the usefulness of the TSs conducted by the PHC, and IC programs that the facilities intended to implement or develop in the future. This study examined the relationship between the perceived usefulness of the information provided and the IC programs planned for development, referencing a study conducted in 2023 and other case reports. Results Seventeen hospitals and six AFs with beds responded to the survey, yielding an 85.2% response rate. IC policies for methicillin-resistant Staphylococcus aureus (MRSA) were prepared by 21 hospitals (91.3%), whereas only five hospitals (21.7%) had prepared IC policies for carbapenem-resistant Enterobacteriaceae. Regarding HHCPs, an increase in the availability of alcohol-based hand sanitizer was identified by 17 hospitals (73.9%), while 13 hospitals (56.5%) reported using posters or symbols, 12 hospitals (52.2%) reported using TS and hand sanitizers, and nine hospitals (39.1%) assessed HH compliance and provided feedback. Furthermore, nine hospitals (39.1%) identified HHCPs and Environmental Cleaning (EC) for carbapenemase-producing Enterobacteriaceae (CPE) as useful information. There was a statistically significant association between TSs on HHCPs and the development of new HHCPs (p = 0.027). Additionally, information on EC for CPE was significantly associated with the development of staff cohorting strategies (p = 0.007). However, TS programs were not significantly connected to EC, nor were TSs to be developed. Conclusion The PHC should advise hospitals to assess if their HHCPs effectively contribute to improving HH compliance. It is essential for the PHC to furnish hospitals with resources and information that aid in the development of EC training. Additionally, the PHC should support the creation of specific and effective TS programs focused on EC or TS development. Conducting surveys to identify barriers to implementing staff cohorting strategies is also recommended. We propose that TS programs should include quantifiable data on HHCPs and EC, such as.

Smoking cessation after cancer diagnosis reduces the risk of severe cancer pain: A longitudinal cohort study.

BACKGROUND: Whether abstinence from smoking among cancer patients reduces cancer pain is still unclear. Opioids can act as a surrogate index for evaluating the incidence of severe cancer pain in countries where opioid abuse is infrequent. This study aimed to investigate whether changed smoking behavior after cancer diagnosis influences the incidence of severe cancer pain as determined by strong opioid use. METHODS: Using a large Japanese insurance claims database (n = 4,797,329), we selected 794,702 insured employees whose annual health checkup data could be confirmed ≥6 times between January 2009 and December 2018. We selected 591 study subjects from 3,256 employees who were diagnosed with cancer pain and had health checkup data at the year of cancer pain diagnosis. RESULTS: A significantly greater proportion of patients who continued smoking after cancer diagnosis ("current smoker", n = 133) received strong opioids (36.8%) compared with patients who had never smoked or had stopped before cancer diagnosis ("non-smoker", n = 383, 20.6%; p<0.05) but also compared with patients who had quit smoking after cancer diagnosis ("abstainer:", n = 75, 24.0%; p<0.05). In multivariable Cox proportional hazards regression analysis, abstainers had a significantly lower risk of receiving strong opioids than current smokers (hazard ratio: 0.57, 95% CI: 0.328 to 0.997). These findings were consistent across multiple sensitivity analyses. CONCLUSION: Our study demonstrated that patients who quit smoking after cancer diagnosis have a lower risk of severe cancer pain. This information adds clinical incentives for improving quality of life among those who smoked at the time of cancer diagnosis.

Characterisation of a novel KRAS G12C inhibitor ASP2453 that shows potent anti-tumour activity in KRAS G12C-mutated preclinical models.

BACKGROUND: KRAS is one of the most frequently mutated oncogenes in various cancers, and several novel KRAS G12C direct inhibitors are now in clinical trials. Here, we characterised the anti-tumour efficacy of ASP2453, a novel KRAS G12C inhibitor, in preclinical models of KRAS G12C-mutated cancer. METHODS: We evaluated the in vitro and in vivo activity of ASP2453, alone or in combination with targeted agents and immune checkpoint inhibitors, in KRAS G12C-mutated cancer cells and xenograft models. We also assessed pharmacological differences between ASP2453 and AMG 510, another KRAS G12C inhibitor, using an SPR assay, washout experiments and an AMG 510-resistant xenograft model. RESULTS: ASP2453 potently and selectively inhibited KRAS G12C-mediated growth, KRAS activation and downstream signalling in vitro and in vivo, and improved the anti-tumour effects of targeted agents and immune checkpoint inhibitors. Further, ASP2453 had more rapid binding kinetics to KRAS G12C protein and showed more potent inhibitory effects on KRAS activation and cell proliferation after washout than AMG 510. ASP2453 also induced tumour regression in an AMG 510-resistant xenograft model. CONCLUSIONS: ASP2453 is a potential therapeutic agent for KRAS G12C-mutated cancer. ASP2453 showed efficacy in AMG 510-resistant tumours, even among compounds with the same mode of action.

The emergence of nebulin repeats and evolution of lasp family proteins.

The LIM and SH3 domain protein (lasp) family, the smallest proteins in the nebulin superfamily, consists of vertebrate lasp-1 expressed in various non-muscle tissues, vertebrate lasp-2 expressed in the brain and cardiac muscle, and invertebrate lasp whose functions have been analyzed in Ascidiacea and Insecta. Gene evolution of the lasp family proteins was investigated by multiple alignments, comparison of gene structure, and synteny analyses in eukaryotes in which mRNA expression was confirmed. All invertebrates analyzed in this study belonging to the clade Filasterea, with the exception of Placozoa, have at least one lasp gene. The minimal actin-binding region (LIM domain and first nebulin repeat) and SH3 domain detected in vertebrate lasp-2 were found to be conserved among the lasp family proteins, and we showed that nematode lasp has actin-binding activity. The linker sequences vary among invertebrate lasp proteins, implying that the lasp family proteins have universal and diverse functions. Gene structures and syntenic analyses suggest that a gene fragment encoding two nebulin repeats and a linker emerged in Filasterea or Holozoa, and the first lasp gene was generated following combination of three gene fragments encoding the LIM domain, two nebulin repeats with a linker, and the SH3 domain.

Aryl Hydrocarbon Receptor Signaling Is Functional in Immune Cells of Rainbow Trout (Oncorhynchus mykiss).

The arylhydrocarbon receptor (AhR) is an important signaling pathway in the immune system of mammals. In addition to its physiological functions, the receptor mediates the immunotoxic actions of a diverse range of environmental contaminants that bind to and activate the AhR, including planar halogenated aromatic hydrocarbons (PHAHs or dioxin-like compounds) and polynuclear aromatic hydrocarbons (PAHs). AhR-binding xenobiotics are immunotoxic not only to mammals but to teleost fish as well. To date, however, it is unknown if the AhR pathway is active in the immune system of fish and thus may act as molecular initiating event in the immunotoxicity of AhR-binding xenobiotics to fish. The present study aims to examine the presence of functional AhR signaling in immune cells of rainbow trout (Oncorhynchus mykiss). Focus is given to the toxicologically relevant AhR2 clade. By means of RT-qPCR and in situ hybdridization, we show that immune cells of rainbow trout express ahr 2α and ahr 2ß mRNA; this applies for immune cells isolated from the head kidney and from the peripheral blood. Furthermore, we show that in vivo as well as in vitro exposure to the AhR ligand, benzo(a)pyrene (BaP), causes upregulation of the AhR-regulated gene, cytochrome p4501a, in rainbow trout immune cells, and that this induction is inhibited by co-treatment with an AhR antagonist. Taken together, these findings provide evidence that functional AhR signaling exists in the immune cells of the teleost species, rainbow trout.

In Vitro Differentiation of Chicken Astrocytes: Growth, Morphology, and Protein Expression of Astrocytes in Primary Cultures.

Astrocytes regulate synaptic transmission in the central nervous system. Astrocytes in vivo have "stems" that express glial fibrillary acidic protein (GFAP), intermediate filaments, and peripheral astrocyte processes (PAPs), which contain actin-rich cytoskeletal structures. At the PAPs, the perisynaptic glia contacts and enwraps synapses, and modulates glia-neuronal communication. Cultured astrocytes have been an invaluable tool for studying roles of astrocytes; however, the morphology of mammalian primary astrocytes cultured in conventional medium containing fetal bovine serum (FBS) was similar to that of fibroblasts, and many culture conditions have been developed to generate stellate astrocytes observed in vivo. Avian astrocytes have been prepared from embryonic chick forebrain and maintained at a high cell density in conventional FBS-containing medium as mammalian astrocytes, thus the morphological analysis of chicken astrocytes has not yet been performed. In the present study, we report that the morphology of astrocytes freshly harvested from the forebrain of a chicken embryo in serum-free Neurobasal medium with B-27 supplement and basic fibroblast growth factor (bFGF) is similar to that of the astrocyte morphology in vivo. We also find that astrocytes in this medium express similar levels of GFAP and two actin-binding proteins as astrocytes in conventional FBS-containing medium, although they have different morphologies. Furthermore, we confirmed that cryopreserved astrocytes differentiate faster than freshly harvested astrocytes.

Immune-Specific Expression and Estrogenic Regulation of the Four Estrogen Receptor Isoforms in Female Rainbow Trout (Oncorhynchus mykiss).

Genomic actions of estrogens in vertebrates are exerted via two intracellular estrogen receptor (ER) subtypes, ERα and ERß, which show cell- and tissue-specific expression profiles. Mammalian immune cells express ERs and are responsive to estrogens. More recently, evidence became available that ERs are also present in the immune organs and cells of teleost fish, suggesting that the immunomodulatory function of estrogens has been conserved throughout vertebrate evolution. For a better understanding of the sensitivity and the responsiveness of the fish immune system to estrogens, more insight is needed on the abundance of ERs in the fish immune system, the cellular ratios of the ER subtypes, and their autoregulation by estrogens. Consequently, the aims of the present study were (i) to determine the absolute mRNA copy numbers of the four ER isoforms in the immune organs and cells of rainbow trout, Oncorhynchus mykiss, and to compare them to the hepatic ER numbers; (ii) to analyse the ER mRNA isoform ratios in the immune system; and, (iii) finally, to examine the alterations of immune ER mRNA expression levels in sexually immature trout exposed to 17ß-estradiol (E2), as well as the alterations of immune ER mRNA expression levels in sexually mature trout during the reproductive cycle. All four ER isoforms were present in immune organs-head kidney, spleen-and immune cells from head kidney and blood of rainbow trout, but their mRNA levels were substantially lower than in the liver. The ER isoform ratios were tissue- and cell-specific, both within the immune system, but also between the immune system and the liver. Short-term administration of E2 to juvenile female trout altered the ER mRNA levels in the liver, but the ERs of the immune organs and cells were not responsive. Changes of ER gene transcript numbers in immune organs and cells occurred during the reproductive cycle of mature female trout, but the changes in the immune ER profiles differed from those in the liver and gonads. The correlation between ER gene transcript numbers and serum E2 concentrations was only moderate to low. In conclusion, the low mRNA numbers of nuclear ER in the trout immune system, together with their limited estrogen-responsiveness, suggest that the known estrogen actions on trout immunity may be not primarily mediated through genomic actions, but may involve other mechanisms, such as non-genomic pathways or indirect effects.

Who needs the hotspot? The effect of temperature on the fish host immune response to Tetracapsuloides bryosalmonae the causative agent of proliferative kidney disease.

Proliferative kidney disease (PKD) of salmonids, caused by Tetracapsuloides bryosalmonae may lead to high mortalities at elevated water temperatures. However, it has not yet been investigated how temperature affects the fish host immune response to T. bryosalmonae. We exposed YOY (young of the year) rainbow trout (Oncorhynchus mykiss) to T. bryosalmonae at two temperatures (12 °C and 15 °C) that reflect a realistic environmental scenario and could occur in the natural habitat of salmonids. We followed the development of the parasite, host pathology and immune response over seven weeks. We evaluated the composition and kinetics of the leukocytes and their major subgroups in the anterior and posterior kidney. We measured immune gene expression profiles associated with cell lineages and functional pathways in the anterior and posterior kidney. At 12 °C, both infection prevalence and pathogen load were markedly lower. While the immune response was characterized by subtle changes, mainly an increased amount of lymphocytes present in the kidney, elevated expression of Th1-like signature cytokines and strong upregulation of the natural killer cell enhancement factor, NKEF at week 6 P.E. At 15 °C the infection prevalence and pathogen burden were ominously greater. While the immune response as the disease progressed was associated with a Th2-like switch at week 6 P.E and a prominent B cell response, evidenced at the tissue, cell and transcript level. Our results highlight how a subtle, environmentally relevant difference in temperature resulted in diverse outcomes in terms of the immune response strategy, altering the type of interaction between a host and a parasite.

A role for multiple estrogen receptors in immune regulation of common carp.

Estrogens are important for bi-directional neuroendocrine-immune interaction. They act via nuclear estrogen receptors (ERα and ERß) and/or G-protein coupled receptor - GPR30. We found expression of ERα, ERß and GPR30 in carp lymphoid tissues and head kidney monocytes/macrophages, neutrophils and lymphocytes. Interestingly, ERß is also expressed in some head kidney lymphocytes but not in naive PBLs. Immune stimulation altered the cell type specific profile of expression of these receptors, which depends on both activation and maturation stage. This implies direct leukocyte responsiveness to estrogen stimulation and therefore in vitro effects of 17ß-estradiol (E2) on reactive oxygen species (ROS) production in monocytes/macrophages were determined. Short-time incubation with E2 increased ROS production in PMA-stimulated cells. Results comply with mediation by GPR30, partially functioning via phosphoinositide 3-kinase activation. These results furthermore demonstrate that neuroendocrine-immune communication via estrogen receptors is evolutionary conserved.

ASP5878, a Novel Inhibitor of FGFR1, 2, 3, and 4, Inhibits the Growth of FGF19-Expressing Hepatocellular Carcinoma.

Hepatocellular carcinoma is an aggressive cancer with poor prognosis. Fibroblast growth factor 19, a member of the fibroblast growth factor family, is a ligand for fibroblast growth factor receptor 4. Moreover, it plays a crucial role in the progression of hepatocellular carcinoma. ASP5878 is a novel inhibitor of fibroblast growth factor receptors 1, 2, 3, and 4 that is under development. It inhibits fibroblast growth factor receptor 4 kinase activity with an IC50 of 3.5 nmol/L. ASP5878 potently suppressed the growth of the fibroblast growth factor 19-expressing hepatocellular carcinoma cell lines Hep3B2.1-7, HuH-7, and JHH-7. In the Hep3B2.1-7 cell line, ASP5878 inhibited the phosphorylation of fibroblast growth factor receptor 4 and its downstream signaling molecules as well as induced apoptosis. Oral administration of ASP5878 at 3 mg/kg induced sustained tumor regression in a subcutaneous xenograft mouse model using Hep3B2.1-7. In HuH-7, an orthotopic xenograft mouse model, ASP5878 induced complete tumor regression and dramatically extended the survival of the mice. These results suggest that ASP5878 is a potentially effective therapeutic agent for hepatocellular carcinoma patients with tumors expressing fibroblast growth factor 19. Mol Cancer Ther; 16(1); 68-75. ©2016 AACR.

ASP5878, a selective FGFR inhibitor, to treat FGFR3-dependent urothelial cancer with or without chemoresistance.

FGF/FGFR gene aberrations such as amplification, mutation and fusion are associated with many types of human cancers including urothelial cancer. FGFR kinase inhibitors are expected to be a targeted therapy for urothelial cancer harboring FGFR3 gene alternations. ASP5878, a selective inhibitor of FGFR1, 2, 3 and 4 under clinical investigation, selectively inhibited cell proliferation of urothelial cancer cell lines harboring FGFR3 point mutation or fusion (UM-UC-14, RT-112, RT4 and SW 780) among 23 urothelial cancer cell lines. Furthermore, ASP5878 inhibited cell proliferation of adriamycin-resistant UM-UC-14 cell line harboring MDR1 overexpression and gemcitabine-resistant RT-112 cell line. The protein expression of c-MYC, an oncoprotein, in gemcitabine-resistant RT-112 cell line was higher than that in RT-112 parental cell line and ASP5878 decreased the c-MYC expression in both RT-112 parental and gemcitabine-resistant RT-112 cell lines. Once-daily oral administration of ASP5878 exerted potent antitumor activities in UM-UC-14, RT-112 and gemcitabine-resistant RT-112 xenograft models without affecting body weight. These findings suggest that ASP5878 has the potential to be an oral targeted therapy against urothelial cancer harboring FGFR3 fusion or FGFR3 point mutation after the acquisition of gemcitabine- or adriamycin-resistance.

Germline RET mutation carriers in Japanese patients with apparently sporadic medullary thyroid carcinoma: A single institution experience.

OBJECTIVE: Genetic testing for RET germline mutation can be useful to distinguish whether a patient with medullary thyroid carcinoma (MTC) is genuinely sporadic or hereditary. Conducting a routine preoperative germline RET genetic screening for all patients with MTC has the clinical benefit, i.e., avoidance of unnecessary total thyroidectomy in the selected patients. We sought to clarify the incidence of germline RET mutation carriers in Japanese patients with apparently sporadic MTC and to address the differences in clinicopathological characteristics between true sporadic MTC and hereditary MTC in these patients, all of whom were treated at Kuma Hospital. METHODS: A total of 134 patients with apparently sporadic MTC who underwent surgery between 1996 and 2014 were enrolled. All patients underwent a germline RET gene mutation analysis preoperatively. RESULTS: Germline mutations in RET proto-oncogene were identified in 20 of the 134 (14.9%) apparently sporadic MTC patients. No significant difference in clinicopathological characteristics was observed between the patients with sporadic MTC (n=114) and those with hereditary MTC (n=20) except for the RET gene carriers' younger age at diagnosis and presence of multifocal and bilateral lesions. CONCLUSION: Germline RET mutations were identified in 14.9% of Japanese patients with apparently sporadic MTC. No clearly decisive clinicopathological characteristics was observed to distinguish whether an apparently sporadic MTC case was genuinely sporadic or unconsciously hereditary. For the treatment strategy decision, it is advantageous to conduct a routine preoperative germline RET genetic screening for all patients with MTC, even if their MTC is apparently sporadic.

Effects of Pregnancy on Papillary Microcarcinomas of the Thyroid Re-Evaluated in the Entire Patient Series at Kuma Hospital.

BACKGROUND: An active-surveillance clinical trial of low-risk papillary microcarcinoma (PMC) patients has been performed at the authors' institution, Kuma Hospital, since 1993. Favorable oncological results have been reported. During the trial, a few patients were encountered with PMC that showed enlargement during pregnancy, and these cases have been reported. During pregnancy, a large amount of human chorionic gonadotropin (hCG) having weak thyrotropin (TSH) activity is produced, possibly affecting the progression of PMC. This study investigated how pregnancy and delivery influenced the progression of PMC in the entire active surveillance PMC patient series. METHODS: From 1993 to 2013, 1841 patients with low-risk PMC chose the active surveillance program. Fifty of the 1549 female PMC patients experienced 51 pregnancies/deliveries. To minimize observer variation, a single specialist sonographer re-evaluated the changes in the size of these 50 patients' PMCs before and after the pregnancies/deliveries. RESULTS: Four patients (8%) showed enlargement of PMC by ≥3 mm; one patient (2%) showed a decrease by ≥3 mm, and the remaining 44 patients (45 events, 90%) showed stable disease. None of the patients had a novel appearance of lymph node metastases during pregnancy. Of the four patients with enlargement, two underwent surgery after delivery, and the other two continued the active surveillance, since their tumors did not grow after the delivery. After delivery, the PMC of one of these four patients remained stable, and another showed a decrease in PMC size. To date, six more PMC patients underwent surgery after delivery for reasons other than disease progression due to pregnancy and delivery: two opted out of active surveillance, two were identified with a nodal metastasis during active surveillance after delivery, one had Graves' disease, and one showed enlargement of nodules of the contralateral lobe. CONCLUSIONS: Pregnancy and delivery was associated with an increase in size of PMCs in only 8% of the 51 pregnancies/delivery cases. None of the patients developed nodal metastasis during pregnancy. Thus, a possible future pregnancy does not prevent such patients from undergoing active surveillance, although watchful observation during pregnancy is recommended.

Incidences of Unfavorable Events in the Management of Low-Risk Papillary Microcarcinoma of the Thyroid by Active Surveillance Versus Immediate Surgery.

BACKGROUND: The incidence of papillary microcarcinoma (PMC) of the thyroid is rapidly increasing globally, making the management of PMC an important clinical issue. Excellent oncological outcomes of active surveillance for low-risk PMC have been reported previously. Here, unfavorable events following active surveillance and surgical treatment for PMC were studied. METHODS: From February 2005 to August 2013, 2153 patients were diagnosed with low-risk PMC. Of these, 1179 patients chose active surveillance and 974 patients chose immediate surgery. The oncological outcomes and the incidences of unfavorable events of these groups were analyzed. RESULTS: In the active surveillance group, 94 patients underwent surgery for various reasons; tumor enlargement and the appearance of novel lymph node metastases were the reasons in 27 (2.3%) and six patients (0.5%), respectively. One of the patients with conversion to surgery had nodal recurrence, and five patients in the immediate surgery group had a recurrence in a cervical node or unresected thyroid lobe. All of these recurrences were successfully treated. None of the patients had distant metastases, and none died of the disease. The immediate surgery group had significantly higher incidences of transient vocal cord paralysis (VCP), transient hypoparathyroidism, and permanent hypoparathyroidism than the active-surveillance group did (4.1% vs. 0.6%, p < 0.0001; 16.7% vs. 2.8%, p < 0.0001; and 1.6% vs. 0.08%, p < 0.0001, respectively). Permanent VCP occurred only in two patients (0.2%) in the immediate surgery group. The proportion of patients on L-thyroxine for supplemental or thyrotropin (TSH)-suppressive purposes was significantly larger in the immediate surgery group than in the active surveillance group (66.1% vs. 20.7%, p < 0.0001). The immediate surgery group had significantly higher incidences of postsurgical hematoma and surgical scar in the neck compared with the active surveillance group (0.5% vs. 0%, p < 0.05; and 8.0% vs. 100%, p < 0.0001, respectively). CONCLUSIONS: The oncological outcomes of the immediate surgery and active surveillance groups were similarly excellent, but the incidences of unfavorable events were definitely higher in the immediate surgery group. Thus, active surveillance is now recommended as the best choice for patients with low-risk PMC.