Cancer genome sequencing now make it possible to quantitatively analyze the past exposure history of carcinogenic factors, by single-point mutation signature analysis. By classifying the trans-ethnic large scale gastric cancer genome by mutation signature, east asia-specific carcinogenic factors of gastric cancer and their interactions with genetic factors were clarified. In addition, by whole genome sequencing of our gastric cancer mouse model, its mutation signatures and driver genes, were shown to be very similar to human gastric cancer, showing its validity as cancer model.

Carcinogenic substances collected in the Percellome Toxicogenomics Database were analyzed for their epigenetic characteristics. Murine liver gene expression profiles of N-ethyl-N-nitrosourea (ENU), Diethylnitrosamine (DEN), 3-Methylcholanthrene (3MC), 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), Pentachlorophenol (PCP), Phenobarbital sodium (PB), Arsenite sodium (As), Carbaryl (Cb), Carbon tetrachloride (CCl4), Azacytidine (Aza), etc. were analyzed using homemade tools and web-based information (including IPA). We will report the relationship of the gene expression and its upstream mechanisms. (Supported by Health and Labour Sciences Research Grant).

We conducted two transcriptomic experiments related to foreign-body carcinogenesis. (1) Colon mesentery of mice after intraperitoneal injection of crocidolite were sampled over time. By the Affymetrix GeneChip, major alteration was monitored after 1 day, i.e., activation of Wnt/catenin system, and predicted involvement of RAS, TGFβ1, etc. (2) Connective tissue surrounding a glass plate, 10x5x1mm in size, implanted subcutaneously to mice were sampled over time. CTNNB1, WNT etc., were suggested to promote cell proliferation.

We would like to discuss on its epigenetics referring to the recent knowledge of foreign body carcinogenesis. (Supported by Health and Labour Sciences Research Grant).

Veklury® (remdesivir, GS-5734TM), the first FDA-approved antiviral for the treatment of COVID-19, is a single diastereomer monophosphoramidate prodrug of an adenosine analogue. It is intracellularly metabolized into the active triphosphate form, which in turn acts as a potent and selective inhibitor of multiple viral RNA polymerases. Remdesivir has broad-spectrum activity against members of the coronavirus family such as SARS-CoV-2, SARS-CoV, and MERS-CoV, as well as filoviruses and paramyxoviruses. The nonclinical safety and efficacy profile of remdesivir will be presented, including results of in vivo efficacy studies, off-target profiling, and repeat dose toxicology studies.

The emergence of the highly pathogenic coronavirus SARS-CoV-2 and its rapid spread leading to the coronavirus disease 2019 (COVID 19) pandemic has posed a serious global public-health emergency. The ongoing pandemic has led to the development of several investigational cell and gene therapy (CGT) products as a potential treatment of COVID-19. The conduct of a clinical trial for an investigational (CGT) product is guided by the Code of Federal Regulations (CFR) Title 21, Part 312, to ensure the safety and rights of subjects in all phases of a clinical investigation. According to 21 CFR 312.23(a)(8), the sponsor is responsible for providing adequate pharmacology and toxicology data to support a conclusion that the proposed clinical trial is reasonably safe to conduct. This presentation will provide an overview of the regulatory considerations for preclinical development programs designed to assess the safety and activity of CGT products for COVID-19.

There are large species differences in sensitivity to exogenous chemicals (xenobiotics). One of the causes of these species differences is species differences in the metabolism of chemicals and their binding to various receptors. In this symposium, we will invite researchers who have been studying species differences in response to xenobiotics and share the latest research results.

In this presentation, I will first outline the species differences in metabolism of cytochrome P450 (CYP) and other phase I and mammalian glucuronosyl transferase (UGT) phase II xenobiotics, including the latest data. We have reported the pseudogeneity of CYP and UGT in predators of animal species. In this talk, as an introduction to the symposium, I will outline which enzymes are pseudogenized in which species and discuss the characteristics of xenobiotics metabolism in each animal species.

Glutathione transferases (GSTs) for insects can influence their sensitivity in insecticides, and as the Lepidoptera comprises major agricultural pests, it is important to study lepidopteran GSTs. We report herein the characterization of a novel GST (GSTU2) in the silkworm, Bombyx mori.

A GSTU2 mRNA was induced in a silkworm strain exhibiting diazinon resistance. The diazinon and GSH conjugation caused by GSTU2 were involved in a substitution of the phosphothioester moiety of diazinon by glutathione. Since this moiety of diazinon is important part for binding to AChE, GSTU2 could play a crucial role in detoxification of diazinon.

Knowing the metabolic capacity of xenobiotics in birds is important for poultry management and ecosystem conservation. In previous studies, we have analyzed the sequences and expression levels of avian cytochrome P450s (CYPs) and UDP-glucuronosyltransferases (UGTs), and hepatic metabolic activities. As a result, it was indicated that the expression pattern of CYP1-3 genes in the liver differed among birds. In addition, the counts of UGT1 genes tended to be smaller and the glucuronide conjugation activity also tended to be lower in carnivorous birds. In this presentation, I will introduce the species difference and evolution of xenobiotic metabolism in birds.

Animals must avoid absorbing toxins. Animals have a sense of bitter taste as a mechanism to avoid ingestion of poisons prior to the absorption. In mammals, the TAS2R family that is expressed in the taste cells of the oral cavity functions as bitter taste receptor proteins, which recognize poisonous substances in food, resulting in neural signals of bitterness. Mammals that have undergone adaptive radiation since the late Cretaceous period have evolved into a variety of dietary species groups. Mammals that rely on plants for diet are more likely to be exposed to toxic plant secondary metabolites. They need to accept a wider range of toxins as bitter. Therefore, plant-eating mammals have dramatically increased the number of TAS2R family genes in their genomes, increasing their repertoire of recognizable toxins. We humans also have one of the largest TAS2R families among all mammals, which is related to the evolution of the arboreal plant-eating primate ancestor of us. This expansion of the TAS2R family is also linked to the evolution of toxic metabolizing enzymes, such as the CYP family expressed in the liver, which has also expanded in plant-eating mammals. Initially, the TAS2R family was discovered as the expressed genes in taste cells of the oral cavity as bitter taste receptors, but subsequent studies have shown functional expression in extraoral tissues, such as the epithelium of the gastrointestinal tract. They may be involved in the regulation of digestion and metabolism, both directly and arthroscopically. Recent developments in next-generation DNA sequencing technology have made it possible to comprehensively examine the genomes and gene expression of many non-model mammalian groups. This presentation will introduce the methodology and findings of comprehensive comparative genomic and gene expression analyses of mammalian groups with diverse dietary habits from the perspective of bitter taste receptor evolution.

Cynomolgus macaques, important non-human primates used in drug metabolism and toxicity studies, show drug metabolism patterns generally similar to humans. However, some differences are occasionally noted due to species differences in drug-metabolizing enzymes such as cytochrome P450 (CYP) including CYP2C9, CYP2C19, and monkey-specific CYP2C76. Similarly, we also found some differences in UDP-glucuronosyltransferase (UGT), including UGT1A6 and UGT1A9, possibly accounting for species differences of UGT metabolism recently noted between monkeys and humans in drug metabolism studies. The results of other drug-metabolizing enzymes will be also presented.

Formation and maintenance of blood and lymphatic vessels are regulated by various cytokines including bone morphogenetic proteins (BMPs) that are members of transforming growth factor (TGF)-β family. Lymphatic vessels (LVs) play critical roles in the maintenance of fluid homeostasis and in pathological conditions, including cancer metastasis. Although mutations in ALK1, a member of the TGF-β/ BMP receptor family, have been linked to hereditary hemorrhagic telangiectasia, a human vascular disease, the roles of activin receptor-like kinase 1 (ALK-1) signals in LV formation largely remain to be elucidated. We show that ALK-1 signals inhibit LV formation, and LVs were enlarged in multiple organs in Alk1-depleted mice. These inhibitory effects of ALK-1 signaling were mediated by BMP-9, which decreased the number of cultured lymphatic endothelial cells. Bmp9-deficient mouse embryos consistently exhibited enlarged dermal LVs. BMP-9 also inhibited LV formation during inflammation and tumorigenesis. BMP-9 downregulated the expression of the transcription factor Prox1, which is necessary to maintain lymphatic endothelial cell identity. Furthermore, silencing Prox1 expression inhibited lymphatic endothelial cell proliferation. Our findings reveal a unique molecular basis for the physiological and pathological roles of BMP-9/ALK-1 signals in LV formation. In this symposium, our current approaches to study the mechanisms how pulmonary LVs are maintained by TGF-β family signals will be discussed.

Pulmonary fibrosis is characterized by progressive fibrosis leading to respiratory failure. Our research group has identified a missense mutation in SFTPA1 as a causative mutation in familial pulmonary fibrosis, which impairs the secretion of SFTPA1 protein and is considered to be the starting point of the disease. Functional analysis using Sftap11-KI mice revealed that necroptosis of type II alveolar epithelial cells is the initial cellular event of the disease. In this presentation, we would like to discuss how necroptosis leads to pulmonary fibrosis by using Sftpa1-KI mice.

Nanomaterials are widely used in various fields. Although the toxicity of carbon nanotubes (CNTs) in pulmonary tissues has been demonstrated, the toxicological effect of CNTs on the immune system in the lung remains unclear. In this study, exposure to Taquann-treated CNTs (T-CNTs) was performed using aerosols generated in an inhalation chamber. At 12 months after T-CNT exposure, fibrotic lesions were enhanced. In vivo and in vitro experiments demonstrated that T-CNT exposure stimulates upregulation of metalloproteinase-12 (MMP12) of alveolar macrophages to contribute to pulmonary fibrosis.

Traditionally, the toxicological target of inhaled formalin (FA) gas was the nasal cavity, and the authorities said that FA will be trapped there and never reaches the lung tissue. However, when we exposed mice to FA gas at a low “sick building syndrome” level of concentration, clear transcriptomic effect of lung and liver were observed. This finding strongly suggested inhalation toxicology should follow what we learn from anaesthesiology, i.e. the end-tidal anaesthetic concentrations reflects the blood concentration in equilibrium.

We will report the lung and liver transcriptomic data on some volatile chemicals and discuss on the mechanistic prediction of pulmonary and systemic toxicity. (Supported by Health and Labour Sciences Research Grant).

Systemic administration of an adenovirus (Ad) vector often causes hepatotoxicity. The E1 gene, which is crucial for the transcription of other Ad genes, is deleted from Ad vector genome. Theoretically, Ad genes should not be expressed by deletion of the E1 gene; however, Ad genes are slightly expressed from Ad vector genome, resulting in hepatotoxicity. In order to suppress the leaky expression of Ad genes, complementary sequences for liver-specific miR-122a were inserted into the 3’-untranslated region (UTR) of the Ad genes in the Ad vector genome. The Ad vector containing miR-122a-targeted sequences in the 3’-UTR of the E4 gene mediated efficient transduction without severe hepatotoxicity.

In recent years, oligonucleotide therapeutics have been attracting attention as a new modality to treat intractable diseases and genetic disorders. Oligonucleotide therapeutics differ from conventional drugs that target proteins in that they are able to regulate the gene expression at the RNA level, a feature that has led to their rapid development and clinical use over the past several years. In this symposium, I would like to outline the basics of oligonucleotide therapeutics and discuss methods to evaluate the activation of innate immunity by oligonucleotide therapeutics.

Immunogenicity (the development of anti-drug antibodies) is an important concern for both the safety and efficacy of engineered protein therapeutics. The accurate estimation of immunogenic potential and appropriate de-immunization are required in non-clinical development. This talk will present overview of clinical immunogenicity and impact, current situation of non-clinical assessments of immunogenicity, and examples of de-immunization of antibody therapeutics.

Owing to mesenchymal stem cells (MSCs) immunosuppressive, angiogenic, and tissue regeneration abilities and multilineage differentiation potential, MSCs are being used in cell therapy and regenerative medicine. On the other hand, there are many unclear points about the mode of action of treatment using MSCs. It is very difficult to analyze which function of MSCs the required efficacy depends on, and at present, there is no internationally established method for MSCs quality evaluation. Since there are many parts that do not apply to the concept of pharmaceuticals, it is necessary to establish an evaluation concept that is specific to stem cell processed products.

For development of next generation vaccines including COVID-19 vaccines, vaccine adjuvants targeting PRRs, which is related to the innate immune system, have shown great promise in their ability to enhance responses to component vaccines. However there is also a risk to induce massive unexpected inflammation by using these adjuvants. Thus it is important to carefully select the correct pre-clinical model to evaluate the safety and efficacy of the adjuvanted vaccines.

In this presentation, we will introduce our recent data regarding development of vaccines and adjuvants in non-human primate model and discuss about how we can use this model more efficiently for immunotoxicology assessment in future.

Cardiovascular disease is one of the main causes of death in developed countries. Cadmium is a heavy metal known as a risk factor for vascular diseases. Metallothionein (MT) is a cytoprotective protein induced by heavy metals such as cadmium and zinc. However, endothelial MT are not induced by inorganic zinc, suggesting that the mechanisms underlying the MT induction are different between endothelial cells and other cell types. In this symposium, we introduce our findings on the role of endothelial MT in cadmium toxicity and the mechanisms underlying the MT induction.

Parkinson’s disease (PD) is a progressive neurodegenerative disease with motor symptoms due to a loss of nigrostriatal dopaminergic neurons and with non-motor symptoms. Although present nosotropic treatments improve the motor disability in PD, the main challenge remains to develop of neuroprotective or disease-modifying treatments. Astrocytes exert neuroprotective effects by production of antioxidants and neurotrophic factors. In this symposium, we will outline a new therapeutic strategy of dopaminergic neuroprotection by targeting antioxidative molecules metallothioneins in astrocytes.

Amyotrophic lateral sclerosis (ALS) is a motor neuron disease with muscle atrophy and respiratory failure. Superoxide dismutase-1 (SOD1) is deposited as aggregates in motor neurons and glial cells of ALS. While natively folded SOD1 binds Cu/Zn that confer structural stability to the protein, SOD1 aggregates do not contain these metals. Thus, dissociation of Cu/Zn ions from SOD1 is required for its aggregation. Metallothionein-I (MT-I) is a Cu/Zn binding protein involved in the homeostasis of these ions. We hypothesize that MT-I has the potential to inhibit pathological conversion of SOD1 into aggregation. In this symposium, we will talk about that MT-I inhibits SOD1 aggregation at molecular and cellular levels.

Cysteine-bound sulfane sulfur in proteins has received much attention as a key factor in cellular redox homeostasis. With an established method to quantify protein-bound sulfane sulfur, metallothionen (MT) isoforms were found to be highly modified by sulfane sulfur with C-S-S-Zn structure which is supported by Raman spectroscopy and a 3D structure modeling analysis. Evaluation of redox-dependent zinc and sulfane sulfur binding in MT3 suggest that oxidation of zinc/sulfur cluster in MT3 yields intramolecular tetrasulfide bridges coupled to zinc release from the protein. In the symposium, we will introduce the function of sulfane sulfur in the redox regulation of MT proteins.

Normal animals have been commonly used in toxicity studies, but recently the development of animal models for human diseases has been remarkably progressed, and model animals are used for toxicity studies, especially in new modalities. Since there could be differences in responses to test articles between normal and pathological model animals, findings in model animals need to be carefully extrapolated to human. In this presentation, I will introduce case studies of translational research for toxicity findings in disease model animals, which were observed in pharmacological studies.

We exemplify a 5-HT₄ receptor agonist, to compare the Cardiovascular (CV) effects between nonclinical and clinical studies. In the nonclinical studies, the 5-HT₄ receptor agonist showed weak or no effects in in vitro assays, or on in vivo electrocardiograms or blood pressure, whereas dose-dependent increase in heart rate (HR) was noted. Although no CV effects were found in a phase I clinical trial, slight increase in HR was seen in the thorough QT study. The HR increase might be due to on-targeted action to the 5-HT₄ receptor.

The deep prediction of the CV risk from nonclinical studies into human is very important to be carried out, by considering MoA or comparing that with reference compounds, etc.

In the patients treated with BACE (beta-site amyloid precursor protein cleaving enzyme) inhibitor Atabecestat, elevation of live enzymes was observed after dosing. In immunohistochemistry using liver biopsy sample from one patient induced liver injury, an adaptive, T-cell mediated response was noted. Furthermore, in using PBMC treated with Atabecestat and its metabolites, IFN-gamma and IL-13 increased. Taken together, one of the mechanisms of Atabecestat induced hepatotoxicity is seems to be immune mediated. In rats 4-wk toxicology study, necrosis of hepatocyte and cellular infiltration and/or hypertrophy of hepatocyte with high values of aspartate aminotransferase (AST), alanine aminotransferase (ALT) were occurred at higher doses. On the other hand, in dog 4-wk study, only slight elevation of AST and ALT without histopathological changes was observed at a highest dose. We recognized Atabecestat has potential of hepatotoxicity induced, highly covalent-binding and active metabolite. However, because there were species different and large safety margin, we decided to develop. The exposure of clinical dose that showed hepatotoxicity was still enough safety margin in non-clinical toxicology studies, but in fact, hepatotoxicity was occurred in clinical study. Because hepatotoxicity induced by Atabecestat may be considered immune mediated, so, it may be difficult to estimate hepatotoxicity in clinical study. However, as a toxicologist, we really carefully use safety margin and should try to investigate and reveal toxicology mechanism in non-clinical study under the development.

MAS-related G protein coupled receptor-X2 (MRGPRX2) is a receptor expressed in human mast cells and associated with drug-induced pseudo-allergic reactions. In our experiments, dog MRGPRX2 was identified as the orthologue of human MRGPRX2, dog or human MRGPRX2-expressed cells could respond to histamine-releasing agents, and the sensitivity of the dog cells to those agents was higher than the human cells. MRGPRX2-expressed cells were considered useful to investigate the relevance of the reactions seen in dogs receiving a new compound to humans and the species difference of the sensitivity.

Cell therapy products (CTPs) are a new pharmaceutical modality containing living cells. The risk of tumorigenesis derived from cells in the product is one of the major concerns specific to CTPs, and appropriate test methods must be developed as needed. A variety of highly sensitive detection methods for tumorigenic cells as impurities of CTPs have been developed in Japan, some of which have been validated through a public-private joint research (MEASURE). The need for global consensus on the tumorigenicity assessment of CTPs is also discussed at an international consortium (HESI CT-TRACS).

In the aspect of nonclinical safety assessment for human cell therapy products, assessment of general toxicity and tumorgenicity, risk assessment of non-cell components and impurities are required to support clinical trials. In the case of genetically modified cell therapy products or cell products using genome-editing technology, sponsors can refer to “Ensuring the Quality and Safety of Gene Therapy Products” (PSEHB/MDED Notification No.0709-2) and the report of gene therapy products using current genome-editing technology by the science board of PMDA (Human Gene Therapy 2020; 31: 1043-53). In this presentation, our nonclinical considerations for cell therapy products will be provided.

As an AMED supported project, we are developing cancer immunotherapy using immune cells derived from HLA haplotype-identical allogeneic iPS cells. Specifically, we are working on the development of iCAR-ILC / NK therapy using NK cells, which are classified as innate lymphoid cells. At this symposium, I would like to introduce the basics of iCAR-ILC / NK therapy and our efforts toward clinical trials including non-clinical safety tests.

Cell therapies have extraordinary promise - a living drug with the ability to localize, proliferate, activate or differentiate in ways that would be impossible for more passive therapeutics. To date more than 1,500 cell therapies have entered over 9,000 clinical trials around the world. Here, we collect the safety data that have been released from these trials to date and analyze the real world evidence to see what clinical hazards are most prevalent and most impactful. This gap analysis then informs our discovery toxicology strategy for the preclinical detection and de-risking for the next generation of cell therapies.

Spray-freeze-drying (SFD) technique is suitable to prepare porous powders for inhalation. Hydrophobic amino acids were effective to obtain SFD powders with both a favorable inhalation performance and antihygroscopic property. We prepared naked pDNA powders by SFD technique with various excipients and a pDNA encoding firefly luciferase. The powder including low molecular weight hyaluronic acid (LHA) as an excipient showed high gene expression and low cytotoxicity. In-vitro inhalation characteristics were significantly improved by the addition of dispersing agent such as L-phenylalanine.

Pulmonary delivery of drugs has become an attractive target and of great interest in the medication as the lung is capable of absorbing pharmaceuticals either for local deposition or for systemic delivery. Targeted drug delivery to lung alveolar region is demanded for the treatment of COVID-19. In this presentation, we will outline the recent advances in mass spectrometry imaging for visualizing the distribution of administered drugs and their metabolites in tissue sections. Then, we will introduce a newly developed inhalation exposure system and share our knowledge obtained in our recent investigations. We hope that our presentation would contribute the efficient development of useful drugs.

Inhalation poisoning is caused by gaseous causative agent. In this presentation, clinical cases of inhalation poisoning encountered in our daily practice in our emergency department.

Carbon monoxide poisoning is the most common gas poisoning encountered in daily emergency medical practice. Especially, when the method of suicide by charcoal briquettes was spread on the Internet around 2005, the number of cases increased nationwide. There are many clinical questions with treating carbon monoxide poisoning, such as indication of hyperbaric oxygen therapy and treatment of late-onset encephalopathy.

Around 2007, a method of generating hydrogen sulfide was spread on the Internet, and many cases of poisoning occurred. The social impact of the fact that it was possible to generate a gas with a toxicity equivalent to that of chemical terrorism using household products was notable, and it became a social problem with many cases of suicide and secondary injury to rescuers.

As a rare situation, oral ingestion of sodium azide, lime sulfur, and arsenic could generate toxic gas through a reaction with gastric acid. It could cause secondary hazard in an emergency department with the patient as the source, which could be a pitfall in treatment.

Other presentations include poisoning from helium gas, respiratory failure caused by inhalation of waterproof spray, carbon dioxide poisoning from dry ice, and poisoning cases from thinner and LP gas inhalation.

Microorganisms exist in every environment on earth, including the oceans and soil, and they construct complex microbial ecosystems based on sophisticated interactions between different species or the same species of microorganisms. Our bodies are no exception, with a huge number of microorganisms, so-called human microbiota, in our digestive tracts, such as the oral cavity and intestinal tract. The human microbiota interacts with the host to build an ecosystem with a certain robustness. While a well-balanced human microbiota contributes to the maintenance of host health through the production of useful metabolites, an imbalance of microbiota can lead to various disorders through the production of disease-related metabolites. In this presentation, we will introduce the relationship between the human microbiome and host health and diseases, including our recent findings based on a metabologenomic approach and the latest reports related to gut microbiota and toxicology.

The human oral cavity is colonized by numerous and diverse microorganisms, which constitute a complex indigenous microbiota comparable to that of the intestinal tract. Recent studies propose that a dysbiotic shift of overall oral microbiota are involved in development of the oral diseases. Structure and dynamics of oral indigenous microbiota are required to be understood accurately and in detail.

We will overview the structure and dynamics of oral indigenous microbiota in health and diseases based on our data and recent literatures in this lecture.

Gut microbiota development affects infants’ and subsequent host health. Bifidobacteria are among the first to colonize the infant gut; however, how they dominate in the ecosystem are poorly understood. Here, we found that the ability of bifidobacteria to utilize fucosyllactose (FL) and its presence in breast milk are important factors affecting infant gut ecosystems. A novel, uncharacterized ABC transporter was a key factor for FL utilization. Further, we found that bifidobacteria possessing the transporter induce elevated acetate concentration, which have been shown to affect infant health.

The gut microbiota influences liver function through intestinal absorption and the subsequent transfer of their metabolites to the liver. This relationship, called “gut-liver axis”, is therefore important in liver diseases. Hepatic encephalopathy (HE) is an end-stage clinical feature of liver cirrhosis (LC) caused by hyperammonaemia and the resulting neuropsychiatric disorders. Ammonia, a neurotoxic substance that can cross the blood-brain barrier, is produced abundantly by gut microbiota. Excess ammonia is normally converted into urea in hepatocytes. However, liver malfunction and the portosystemic shunts in LC patients cause a direct influx of ammonia into the systemic circulation, resulting in hyperammonaemia and neuropsychiatric disorders. However, species causing hyperammonaemia in HE patients remain unknown. In this study, we aimed to identify specific gut microbial species responsible for hyperammonaemia in HE patients, focusing on the responses to a non-absorbable antibiotic, rifaximin (RFX). We compared the faecal gut microbial profiles of healthy control and HE patients by 16S rRNA gene amplicon sequencing. We used linear discriminant analysis effect size (LEfSe) to identify species, and verified their function in vivo in mouse LC model. We found that urease-positive gut bacterial species, which is known to originally reside in oral cavity, is responsible for hyperammonaemia in RFX-responders, suggesting that these bacterial species transfer to the intestine through breaking stomach acid barrier due to the medication of proton pump inhibitors.

Organisation for Economic Co-operation and Development (OECD) is the international body which has played a central role in harmonizing GLP activities. OECD GLP Working Party, consisting of delegations from member countries, performs various activities including the publication of guidance and discussion of emerging technologies to maintain and promote the harmonization. The COVID-19 has significant impact on GLP activities and inspections in the OECD countries. In the session, how OECD countries react to the pandemic in addition to updates of domestic and overseas GLP issues will be discussed.

If you are using a microscope in your GLP business, quality control and quality assurance of the microscope itself is also an important point. In addition, all image data from microscopes are now recorded as digital data. Therefore, users of microscopes must be careful in handling the image data. In this presentation, I will explain the points that should be recorded in microscope inspection in an easy-to-understand manner.

Pandemic of COVID-19 requires us to reduce physical contact and it accelerates using remote work instead of traditional working style at office. The field of clinical and non-clinical study is no exception. Under this situation, AI is a powerful tool to realize the remote work. LPIXEL has already developed 4 approved medical programs, EIRL Brain Aneurysm, EIRL Brain metry, EIRL Chest Nodule and EIRL Chest metry. More than 100 hospitals/clinics have already installed those 4 programs, and it means our programs is not proof of concept but it's timing for social implementation. Though in the field of diagnosis by medical images, teleradiology is getting popular as a method to work remotely, approved medical programs are expected as a method not only to avoid missed diagnosis but also to make doctors free from time limitation.

LPIXEL also tries to collaborate with several pharmaceutical companies and accelerate drug discovery by implementing AI. Our technology has been already implemented in the discovery phase and our target expand as pre-clinical phase.In the test of safety test of micronucleus, because our AI brings 0.85 correlation with visual inspection, it is expected to reduce test time. By expanding our AI as GLP grade, typical micronucleus can be found by AI and atypical ones are found by expert then, AI play important role to reduce total work time.

AI is promising to apply not only medical fields but also drug discovery, include the field of pre-clinical study. In this presentation we show our product as the case of social implementation and its possibility to expand at pre-clinical fields.

Japan Society of Quality Assurance GLP Division has discussed computerized system and quality assurance of electronic data in test facilities for promotion of electronization in non-clinical testing operation. In recent years, it has proposed the optimum methods for management and quality inspection by analyzing the guidance on management of computerized system by OECD, guidance on Data Integrity by MHRA.

Based in these discussions, we will explore the issues and solutions in the view of Data Integrity regarding handling of approvals of electronic records and utilization of external IT resources such as clouds, which seems important in home working, assumed in non-clinical test. Furthermore, we will refer to remote inspection, which is increasing in contract studies.

We conducted a questionnaire to pharmaceutical companies and CROs about adaptation to the "New Normal" era caused by the corona pandemic. The answers revealed that about 80% of facilities were affected by this environmental changes. Particularly, the way of working are changing to reduce opportunity of physical contact to people. In this presentation, we would like to introduce the management of each company for the "New Normal", and give a perspective on the issues that were seen from the answers and the systems that is desired to be constructed in the future.

We have been exposed to foreign chemicals through ingesting foods, which contain several bioregulatory components showing preventive effects on aging and diseases. It is known that food components produce various reactive intermediates through metabolism and non-enzymatically modify biomolecules to form modified adducts on the molecule. Although protein modification by food components is expected to be involved in the "food functionality", most of its molecular mechanism remains unclear. This time, we introduce our research focusing on vitamin C and polyphenol-modified proteins.

Lysine acylation of histones is a post-translational modification that regulates epigenetic gene expression. Acyl modification on lysine residues is induced by endogenous carboxylic acids such as fatty acids through in vivo chemical reactions. We are exposed daily to various exogenous carboxylic acids with foods, which may be added to lysine residues of target proteins including histone. In this presentation, we will introduce the regulation of gene expression by a food preservative sorbic acid through histone sorbylation and discuss possible influences of exposure to carboxylic acids in food from the perspective of lysine adduct exposome.

Exposome, which is a concept referring to total of lifetime exposure, is divided into three categories, general external, internal, and specific external. We have focused on electron acceptors, which generate reactive oxygen species (ROS), in the specific external exposome and have found that exposure to 9,10-phenanthraquinone (PQ), an electron acceptor, activates PTP1B/EGFR signaling through S-oxidation of PTP1B. This result lets us an assume that combined exposure to electron acceptors enhance ROS-mediated cellular responses such as EGFR signaling compare to individual exposure. Herein, we would like to introduce our research about PQ and discuss such a specific external exposome study.

There is a concept of exposome that elucidates the whole picture that various environmental stresses give to human life. In the enormous environmental stress, proteins often intervene when the environment affects the living body. We are trying to elucidate “Adduct Exposome”, which are limited exposomes focusing on the phenomenon that these environmental molecules directly modify amino acid residues of proteins through in vivo reactions. We introduce recent proteomics using mass spectrometry and information science and introduce an approach to “Adduct Exposome”.

Alkyne is a tiny functional group, which can be combined with various functional groups by click reaction, and now widely used as a tag for small compounds. Alkyne is also known to have a strong Raman signal in the cellular silent region, in which no endogenous molecules show Raman signal. Therefore, we applied Raman microscope for the detection of alkyne-tagged molecules and successfully established alkyne tag Raman imaging (ATRI). Recently, we also applied ATRI for the analysis of protein modifications by small compounds and developed a new method, alkyne-tag Raman screening (ATRaS). In this presentation, several examples of ATRI and ATRaS will be shown.

Recently, revisions to the ICH guidance on the evaluation of toxicity to reproduction (ICH S5(R3)) have been finalized. The revisions expand circumstances under which the outcome of “preliminary EFD (Embryo-Fetal Development) studies” (per ICH M3(R3)) can support clinical trials, which could lead to fewer animal studies being conducted. In addition, the revised guidance provides basic principles that will assist in the development and potential regulatory use of alternative assays for evaluating adverse effects on embryofetal development. This is a new trend for the pharmaceutical safety evaluation.