Regulatory T (Treg) cells, which are expressing the transcription factor Foxp3, are actively engaged in the maintenance of immunological self-tolerance and homeostasis. Depletion or functional impairment of Treg cells is able to enhance cancer and microbial immunity, while their numerical expansion or functional augmentation is instrumental in treating autoimmune disease and establishing graft tolerance. How to achieve these aims by targeting Treg cells with biologicals (such as monoclonal antibodies) or chemicals has been an issue of intense investigation. We have recently shown that certain tyrosine kinase inhibitors that blocked T-cell receptor-proximal signaling in T cells were able to specifically deplete mature Treg cells, thereby enhancing tumor immunity in humans. On the other hand, inhibitors of a serine threonine kinase involved in a T-cell signaling pathway evoked Foxp3 expression in conventional T cells and converted them to functionally competent Treg-like cells, which effectively suppressed autoimmune disease and allergy in animal models. It will be discussed how Treg cells be pharmacologically targeted to control a variety of physiological and pathological immune responses.

Despite the fact that the executive neurocircuitry and neurochemistry for sleep/wake switching has been increasingly revealed in recent years, the mechanism for homeostatic regulation of sleep, as well as the neural substrate for "sleepiness" (sleep need), remains unknown. To crack open this black box, we have initiated a large-scale forward genetic screen of sleep/wake phenotype in mice based on true somnographic (EEG/EMG) measurements. We have so far screened >8,000 heterozygous ENU-mutagenized founders and established a number of pedigrees exhibiting heritable and specific sleep/wake abnormalities. By combining linkage analysis and the next-generation whole exome sequencing, we have molecularly identified and verified the causal mutation in several of these pedigrees. Biochemical and neurophysiological analyses of these mutations are underway. Since these dominant mutations cause strong phenotypic traits, we expect that the mutated genes will provide new insights into the elusive pathway regulating sleep/wakefulness. Indeed, through a systematic cross-comparison of the Sleepy mutants and sleep-deprived mice, we have recently found that the cumulative phosphorylation state of a specific set of mostly synaptic proteins may be the molecular substrate of sleep need.

Recently, many studies on genetic polymorphisms (PGx) in drug transporters and transporter-mediated drug-drug interactions(DDI) have been published, and these are part of mechanisms of interindividual difference in drug response. It is important to predict such interindividual difference at early stage of drug development. With a PBPK model, the effect of changes in transporter function on the Pharmacokinetics (PK),and, ultimately, the Pharmacodynamics (PD) and/or Toxicodynamics (TD) will be predicted. I will focus on the following subjects. Recent reports provided quantitative predictions for OATP1Bs-mediated DDIs between statins and cyclosporine A(CsA)/rifampicin(RIF) based on PBPK models. In the process of the analyses, the in vitro–in vivo discrepancies in the Ki values for OATPs were suggested. I will share with you our approach to make the bottom-up prediction possible. Finally, it should be also challenging to reproduce the inter-individual variabilities by considering the mean value and deviation of each PK parameter in a certain population with PBPK model (so-called "virtual clinical study").

More than 800 human GPCRs allow the selective detection of extracellular signals as diverse as photons, odorants, flavors, nucleotides, hormones, neurotransmitters – revealing GPCRs fundamental role in signal transduction. As they regulate many central physiological processes and are thus implicated in many diseases, GPCRs are among the most important targets for modern medicines. In spite of this medical importance and the recent progress in elucidating the 3D structures of various GPCRs, central questions how these receptors recognize extracellular chemical signals and transfer them across the cellular membrane to finally evoke an intracellular response are largely unresolved at a molecular level, mainly because the different steps during signal transmission are not directly accessible by experiments. In this context we are concentrating on central questions of GPCR mediated cellular signalling using computer based molecular dynamics simulations. Our work revealed for the first time, in atomic detail, the entire process of transmembrane signalling of various GPCRs: we found that ligand binding induces a series of conformational changes within a GPCR which opened a gate inside the receptor for water molecules entering the internal region of the receptor and subsequently driving conformational switches within the receptor which finally led to the activation of a G protein on the intracellular side of the receptor. We have applied our findings successfully to various GPCR targeted drug discovery projects.

Microglia are thought to work as immunocompetent cells in central nervous system. Microglia are also thought to be derived from primitive macrophages in the yolk sac. In normal conditions, microglia are ubiquitously distributed in the spinal cord and brain, and have small cell bodies bearing branched and motile processes, which seem to monitor the local environment. In the pathophysiological condition, microglia are activated, change morphologically, increase in cell number, and alter the expression of genes, including neurotransmitter receptors, such as P2 purinergic receptors. Activated microglia express several subtypes of ionotropic (P2XRs) and metabotropic P2 receptors (P2YRs). Extracellular ATP stimulates microglial P2 receptors to evoke various cellular responses. For example, P2X4Rs play a key role in evoking and maintaining neuropathic pain. P2Y12Rs and P2Y6Rs play very important role for microglial trafficking and phagocytosis, respectively. These findings will open new strategy for drug development. Indeed, the inhibitor of P2X4R has a high potential for being a first in class medicine against neuropathic pain. I think that Pharmacologists are essential for drug discovery.

The mammalian alimentary tract harbors hundreds of species of commensal microbes that critically influence a multitude of host physiological functions. Unfavorable alterations of the gut microbiota composition often correlate with several negative health outcomes. Thus, the amelioration of microbiota dysfunction is a promising route for future therapeutics for several diseases. We have been aiming to understand the features and functions, particularly immunological attributes, of the microbiota, and trying to identify responsible bacterial species and factors for shaping the immune system. We have succeeded in isolation of human and mouse gut-associated commensal bacterial strains that specifically affect the development and function of Th17 cells, Treg cells, Th1 cells or CD8 T cells. In addition, we have identified trypsin-degrading bacterial species. Our findings would allow for designing bacterial consortia that activate or suppress specific adaptive immune programs, potentially resulting in development of better therapeutics for numerous diseases involving the immune system, including infectious disease, autoimmunity, allergy, and cancer.

G protein coupled receptors (GPCRs) comprise the main conduit for information transfer between cells and their environment and participate in most physiological processes through innumerable signaling networks. Accordingly, GPCRs are the largest family of pharmaceutical targets in therapeutic areas ranging from cardiovascular and metabolic diseases to neuropsychiatric and behavioral disorders. Here, I will describe our results from the structural and functional characterization of GPCRs in complex with a range of ligands and signal transducers, providing general mechanisms of transmembrane signal instigation from different receptor classes in this family.

Monoamine transmitters, e.g. dopamine (DA), norepinephrine (NE) and serotonin (5-HT), are playing important roles in a plethora of physiological processes, including reward, attention and mood regulation. Malfunction of the monoaminergic signaling is implicated in diseases such as addiction, stress and depression. Take the DA as an example, a longstanding yet largely unmet goal is to measure DA changes reliably and specifically with high spatiotemporal precision, particularly in animals executing complex behaviors. We developed the new genetically encoded GPCR-activation-based-DA (GRAB_DA) sensors that enable these measurements. GRAB_DA sensors can resolve a single-electrical-stimulus-evoked DA release in mouse brain slices and detect endogenous DA release in living flies, fish, and mice. We have further expanded the repertoire of GRAB sensors for detecting monoamines, purinergic transmitters (e.g. adenosine, ADP and ATP) and neuropeptides (e.g. cholecystokinin, somatostatin and vasoactive intestinal peptide). In sum, these newly developed GRAB sensors provide powerful tools for understanding the regulation and malfunction of monoaminergic and purinergic systems and neuropeptides in both physiological and pathological processes.

Glutamatergic synapses play central roles in almost all of neuronal functions such as learning, motor and sensory functions. Among glutamate receptors, AMPARs are the "actual mediator" at glutamatergic synapses. Despite the accumulation of knowledge of physiological roles of AMPARs, its clinical translation is limited. Main reason for this is that we are not currently able to visualize AMPARs in living human brain. We developed novel PET probe for AMPARs, named [¹¹C]K-2. We detected [¹¹C]K-2 signals reflecting specific binding to AMPARs in rat, non-human primate and human. We detected significant positive correlation between [¹¹C]K-2-signals and protein amount of AMPARs with surgically removed tissue from epileptic patients. Thus, our PET probe for AMPARs specifically detects AMPARs and the first PET probe to visualize AMPAR in living human brain. We are currently imaging patients with neuropsychiatric disorders. Further, we have recently identified CRMP2-binding compound, edonerpic maleate, facilitates synaptic AMPAR delivery and results in the acceleration of motor function recovery after brain damage in rodent and non-human primate.

Inflammation is an integral component of many diseases e.g. arthritis, periodontal disease, asthma, cardiovascular diseases and neurodegenerative diseases. Using a systems approach with self-limited inflammatory infectious exudates to map tissue events, cell traffic and identification of protein and chemical mediators, we uncovered three structurally distinct families of potent omega-3 fatty acid-derived (EPA, DPA, DHA) novel endogenous mediators, named resolvins, protectins and maresins coined specialized pro-resolving mediators (SPM) and recently the biosynthesis of peptidio-SPM within these families that are involved in tissues regeneration (denoted , cys-SPM). Complete structural elucidation and total organic synthesis of each new molecule and pathway intermediates confirmed their roles in vivo in the resolution of inflammation in animal models. Each member of this super-family is structurally distinct and is a pro-resolving mediator controlling the duration and magnitude of acute inflammatory responses with actions in pico-nanogram range in animal disease models. Mapping of these resolution circuits provides new avenues to probe the molecular basis of many widely occurring diseases (CN Serhan, Nature 2014, Molecular Aspects of Medicine 2017, Serhan, Levy JCI 2018). This special presentation will focus on our recent advances in the biosynthesis and functions of SPM and the role of the vagus nerve in controlling infectious inflammation. We've operationalized LC-MS-MS based targeted metabololipidomics to profile SPM and recently cross validated this approach with other laboratories using coded samples from human endotoxin challenges establishing SPM biosynthesis in humans and function (Norris et al 2019). Our recent evidence indicates a new role for the vagus nerve and vagotomy in regulation of lipid mediators. Specifically, vagotomy reduces pro-resolving mediators such as lipoxins, resolvins, protectins and maresins delaying resolution in mouse peritonitis and E. coli infections. The vagus regulates peritoneal Group 3 innate lymphoid cell (ILC3) number and peritoneal macrophage responses with lipid mediator profile signatures showing elevated pro-inflammatory mediators and reduced resolvins, including the novel protective immunoresolvent agonist protectin conjugate in tissue regeneration 1 (PCTR1; Dalli et al., Immunity). Results obtained with human vagus ex vivo indicate that vagus produces both proinflammatory lipid mediators (i.e., prostaglandins and leukotrienes) as well as SPM obtained using targeted LC-MS-MS profiling. Electrical stimulation of human vagus ex vivo reduces both prostaglandins and leukotrienes and increases resolvins and other SPMs. These results elucidate a host-protective mechanism mediated by vagus stimulation. Moreover, they define a new pro-resolution of inflammation reflex operative in mice and isolated human tissue that involves a vagus-SPM circuit. Together these results indicate that endogenous resolution pathways may underlie prevalent diseases associated with uncontrolled inflammation and open the potential for resolution-based physiology, resolution pharmacology and resolution-bioelectric medicine.

Recent advances in stem cell biology may make possible new approaches for the treatment of a number of diseases. A better understanding of molecular mechanisms that control stem cell fate as well as an improved ability to manipulate them are required. Toward these goals, we have developed and implemented high throughput cell-based screenings of chemical libraries, and identified and further characterized small molecules that can control stem cell fate in various systems. This talk will provide latest examples of discovery efforts in my lab that have advanced our ability and understanding toward controlling stem cell fate, including self-renewal, survival, differentiation and reprogramming of stem cells.

Polyunsaturated fatty acids (PUFAs) exhibit a wide range of biological effects, many of which are mediated through the formation and actions of their bioactive metabolites. It is well appreciated that different PUFA balance affects inflammation and related diseases, and recent advances in mass spectrometry-based lipidomics technologies have revealed potential links between PUFA metabolism and biological phenotypes. Omega-3 PUFAs such as EPA and DHA are widely held to be beneficial in maintaining tissue homeostasis. To date, we've developed LC-MS/MS-based lipidomics system to monitor fatty acid metabolites comprehensively, and uncovered the roles of omega-3 PUFA metabolites in controlling inflammatory responses. I'll describe recent advances on omega-3 PUFA–derived mediators, mainly focusing on omega-3 oxygenation pathway that may hold a key for the beneficial effects of omega-3 PUFAs by structure-specific mechanisms in controlling inflammation and tissue homeostasis. Also I'll introduce our recent technology development, namely non-targeted lipidomics, that has a strong potential to search for lipids of interest globally, and to identify unknown lipid species in a non-biased fashion.

G protein-coupled receptors (GPCRs) are the largest family of cell surface drug targets. Consequently, there is high interest in understanding the structure of members of this receptor superfamily and molecular detail of how ligands and transducer proteins interact with them. Our laboratory has been applying single particle cryo-EM to determination of active GPCR structures, using minimally modified receptors. Our work has been principally focused on the class B GPCR subfamily that bind large peptide hormones and are well established clinical targets for the treatment of major disease, including migraine, irritable bowel syndrome, diabetes, obesity and neurodegeneration. We have now solved structures of multiple different receptors, providing wide structural coverage of the major subfamilies of class B GPCRs. Included within this are structures of the same receptor bound to native peptide agonists, biased peptide agonists and non-peptide agonists and receptors in complex with accessory proteins that allosterically modulate receptor function. In combination with molecular pharmacology and molecular dynamics simulations, we are gaining substantial insights into diverse modes of ligand binding, receptor activation and modulation that lead to G protein coupling and downstream signalling.

Memory formation triggers plastic changes in a defined population of neural cells. We elucidated how synaptic inputs activate nuclear CREB-dependent transcriptional activation in these neurons, and uncovered new activity-regulated neural plasticity machineries from the synapses to the nucleus, and then back to synapses. Specifically, we found several CREB-transcription regulating pathways, as well as an inverse synaptic tagging mechanism governed by a CREB target gene Arc, which together help maintain a long-term enhancement of synaptic strength contrast between potentiated and weakened synapses. Elucidation of the genomic and protein structural modules underlying this bidirectional link laid the foundation for rational design of novel neural activity probes E-SARE and XCaMP, useful for imaging of cellular plasticity and neural activity, respectively. Together, these tools are accelerating new research in neural circuit functions and brain information dynamics.

The current therapeutic drugs mainly modulate monoaminergic signaling. Since they are not effective for all patients, the development of novel therapeutic target is required. Recently, it has been reported that inflammation-related molecules are increased in the blood from patients with psychiatric disorders such as major depression. Therefore, neuroinflammation is a possible cause of these disorders. However, we still do not know whether neuroinflammation induces depression. We found that repeated social defeat stress, an animal model of depression, induces behavioral changes, accompanying with microglial activation in the specific brain area. Based on these findings, we revealed the essential role and molecular basis of neuroinflammation. In addition, we developed the drug screening platform which targets neuroinflammation for neurodegenerative disease. In this presentation, I'd like to introduce the recent our findings on neuroinflammation which causes neurological disorders.

Major depressive disorder is one of the most widespread mental illnesses, affecting more than 264 million people worldwide (World Health Organization, 2019) and causing enormous personal and socioeconomic burden. Conventional monoaminergic antidepressants have significant limitations, including delayed onset of therapeutic response and low efficacy: approximately one-third of depressed patients fail to respond to multiple antidepressant treatments and are considered treatment-resistant depression. Recent studies reveal that ketamine, an N-methyl-D-aspartate receptor antagonist, produces rapid antidepressant actions in patients with treatment-resistant depression. However, clinical use of ketamine as an antidepressant is limited due to its serious drawbacks including psychotomimetic/dissociative effects and abuse potential. Thus, there is a continuing unmet need for the development of safer rapid-acting antidepressants. I have demonstrated that resolvins, bioactive metabolites derived from docosahexaenoic acid and eicosapentaenoic acid, produce antidepressant effects through mechanisms (activation of mTORC1 signaling) similar to those underlying the rapid antidepressant effects of ketamine. Taken together, resolvins could be promising targets for the development of novel rapid-acting antidepressants with fewer side effects than ketamine, since resolvins are endogenous lipid mediators that play an important role in homeostasis.

Prostaglandin D₂(PGD₂) is a biological substance that abundantly produced by mast cells. In the development of mast cell-dependent allergic inflammation, a large amount PGD₂is released from activated mast cell. Since there have been reported that PGD₂play both pro- and anti-inflammatory roles in the development of allergy, the precise role of mast cell-derived PGD₂remained unclear. By using systemic and mast cell-specific PGD synthase deficient mice, we demonstrated that PGD₂-DP1 signal inhibited the development of food allergy and anaphylaxis. We also found that the signal enhancement of DP1 exerted both preventive and therapeutic effects in the development of food allergy. Now, we try to reveal the role of PGD₂-DP2 axis in the allergic sensitization. In addition, we develop new diagnosis methods for food allergy, based on our findings that the urinary PGD₂metabolite reflect the severity of symptom. In this presentation, I would like to show our recent results from experimental and clinical research.

Recently, the importance of observational clinical studies utilizing real world data (RWD) is highlighted. In Japan, so far large scale medical databases including claims data, prescription data, and DPC data have been utilized for the clinical epidemiology and pharmacoepidemiology studies, however, outcomes evaluation has been limited due to lacking of laboratory data. Therefore, starting from 2015, we are currently developing large scale electrical medical record (EMR)-derived database in Japan for the clinical epidemiology studies. We also focus on the development of the health checkup databases of newborn, infant, and children population, under the contract with more than 140 Japanese local city governments. These activities and the future opportunities for clinical research and preventive medicine will be discussed.

With respect to study design, randomized control trials (RCT) are considered the gold standard in assessing the efficacy and safety for different medications. However, in addition to the issues of appropriateness, ethics, feasibility, time and costs in RCTs, the generalizability of the findings is argued because of the highly selected group of patients included in RCTs. Real-world studies using a large database, which consists of routinely collected clinical data, are complementary to RCTs in the sense that the patient populations are much more generalizable, and are becoming more common. Using the Diagnosis Procedure Combination (DPC) database, which is a nationwide database and covers approximately 50% of the acute-care inpatients in Japan, we conducted a retrospective cohort study evaluating the efficacy of several drugs including Japanese herbal Kampo medicine. Propensity score utilizing methods were adopted in these studies. Examples of our studies using the DPC database will be presented and the advantages and limitations of the studies will be discussed.

National Database of Health Insurance Claims and Specific Health Checkups of Japan (NDB) is the largest insurance claim database in Japan. The aim of NDB is to analyze health care services provided by health insurance in Japan based on the Act on Assurance of Medical Care for Elderly People (2008). At the end of 2014, approximately 90% of all the health insurance claims were stored in NDB. In the present study, we described the proportion of prescription of drugs listed in Screening Tool for Older Person's appropriate Prescriptions in Japanese (STOPP-J) as those that require careful administration in older adults, using the data from NDB. Participation was limited to those who were aged equal to or more than 75 years and received any medical care provided by health insurance during the 2010-2014 period. Results suggest that more than 60% of the subjects had experiences to be regularly prescribed at least one of the drugs listed in STOPP-J. Although the prescriptions which examined in the present study do not mean inappropriate medications, further considerations for types of drugs might be required when doctors prescribe some drugs for older patients in actual clinical settings.

The Health, Care and Educational Information Evaluation Promotion Organization (HCEI) has established a database centered on electronic medical records in collaboration with 180 medical institutions since 2015. As a characteristic of this database, it contains not only data of DPC and receipt already, but also test results and death data. As a result, we can conduct outcome research to measure the effects of drugs and treatments.

Our database is not dependent on vendors and register in the database about 19 million patients. Based on JLAC10, it has standardized 1000 kinds of inspections (with unit / sample classification), and it becomes a useful database for post marketing surveillance etc.

In addition, we have also started a project to collect data in the registry and the data extraction in the randomized controlled trial conducted by the academic society, and we are promoting efforts to eliminate the load of data extraction in the hospitals. Since effective utilization of medical data is a major cornerstone for drug discovery or quality improvement of medical treatment, we would like to continue to develop the database for the promotion of primary and secondary use of medical information and to save future lives.

Nihon University, having been striving for the further improvement of medical progress and medical services, is engaged in research concerning the utilization of information from daily clinical practice, and in 2004 has constructed the clinical data warehouse (CDW) known as "Nihon University School of Medicine's Clinical Data Management System" (NUSM's CDMS) As of March 2019 for 15years, this system stores over 2.43 million patient profiles, medical history data for 1.27 mil. patients (24 mil. records), drug prescription data of 690,000 patients (40 mil. records), injection medicine prescription data of 270,000 patients (17.5 mil. records), and clinical test data of 800,000 patients (440 mil. records). This system possesses a sufficient amount of information for highly reliable statistical analysis. We have announced to international journals our research thus far that utilizes this abundance of information, which includes studies of the additional effects in hypertensive drugs (add-on effects), and of the side-effects that occur in the combined use of antithrombotic drugs. NUSM's CDMS is a healthcare database based on information from daily clinical practice and furthermore constructed on the premise of use in research, already at a sufficiently practical level. In observational studies using NUSM'S CDMS, the drug effects, or so-called "effectiveness", in actual clinical settings, which cannot be understood through randomized clinical trials (RCTs) carried out in limited environments, are able to be verified and helpful evidence provided.

Chemotherapy-induced peripheral neuropathy (CIPN), a potentially dose-limiting toxicity, impairs the quality of life in cancer patients, whereas there are no effective countermeasures for prevention or treatment of CIPN. Our fundamental studies have demonstrated the critical role of HMGB1, a DAMP molecule, in the development of CIPN following treatment with paclitaxel, vincristine, oxaliplatin, etc. in rodents, and indicated that anti-HMGB1-neutralizing antibodies and thrombomodulin alfa capable of inactivating HMGB1 are useful in inhibiting the development of CIPN. Our retrospective cohort studies in cancer patients undergoing chemotherapy have shown that hepatocellular damage is associated with increased severity of CIPN following oxaliplatin treatment, and that women with breast cancer over menopause age have a higher risk for the incidence of CIPN following paclitaxel treatment. Our animal experiments conducted on the basis of those clinical findings have revealed that experimentally induced hepatocellular damage and ovariectomy aggravate the CIPN caused by oxaliplatin and paclitaxel, respectively, in an HMGB1-dependent manner. Collectively, bidirectional studies between bench and bedside unveil previously unknown risk factors for the incidence and/or increased severity of CIPN, in which HMGB1 might play an essential role.

In recent years, large-scale medical information databases for diseases and their related side effects have been used to accurately assess the effects and side effects of clinically used drugs. Integration of basic and clinical research with medical information databases can enable various analyses based on real-world data. We used this approach to improve the quality of cancer treatment by developing drugs to reduce the side effects caused by anticancer drugs. Focusing on acute kidney injury (AKI) caused by cisplatin (CDDP), we found that several existing drugs suppress the occurrence of CDDP-induced AKI in the FAERS (FDA Adverse Event Reporting System). In vitro and in vivo experiments revealed that administration of candidate drugs extracted by FAERS analysis significantly suppressed CDDP-induced AKI. These results suggest that the existing pharmaceutical products selected using FAERS could prevent CDDP-induced AKI. Effective use of medical data is a major cornerstone for drug discovery and improvement of the quality of life for patients with cancer. This novel strategy will provide a means to expand the possibilities of drug delivery and to elucidate mechanisms of unknown diseases.

Recent pharmacological studies have been developed based on the finding of new disease-related gene, which is accompanied with the production of gene-manipulated disease model animals and high-affinity ligands for the target protein. However, emergence of the gene-based strategy has led to the rapid deprivation of drug target molecules. To overcome this situation, we have been trying to utilize clinical big data to explore a novel and unexpected hypothesis of drug-drug interaction that leads to the drug repositioning. Here, we introduce our data-driven approach in which adverse self-reports, JMDC claims database and University hospital health records are analyzed and compared to find and validate new drug targets. We also present our recent effort to predict the binding affinity of theoretically any chemical ligands to a target protein by deep learning of chemical structures and their measured affinity to the target with graph convolutional neural network. The hypothesis and prediction provided by data-driven approaches will impact on the style of pharmacological study.

Through the infrastructure development of Translational Research Sites with government supports, translational researches that lead basic research outcomes to clinical development have been activated. As a result, pharmaceuticals and medical devices development originating in academia have been vitalized. The drug discovery originating in academia has been active since pharmaceutical companies introduced the idea of open Innovation.

As one of the Translational Research Sites, the Clinical Research, Innovation and Education Center, Tohoku University Hospital (CRIETO) has been coordinating about 10 investigator-initiated clinical trials every year and providing support for over 200 development seeds.

Based on the current situation, I will present the future prospects of drug discovery originating in academia from the standpoint of supporting the seeds development.

Kyoto University has a background of abundant basic research with beautiful science, and research for clinical application is also thriving to return these results to society.

iACT, an academic research organization in Kyoto University Hospital, connects with on- and off-campus researchers, finds potential candidates for clinical application, provides support for appropriate funding and clinical trials.

In recent years, some researchers completed pre-clinical evaluations such as pharmacological tests, toxicity tests, and pharmacokinetic tests of low molecular weight compounds discovered by academia, and manufactured investigational drugs by contract for clinical application. There are also many research proposals related to iPSCs-derived regenerative medicine or repurposing of approved drugs based on screening using iPSCs.

iACT aims to accumulate a wide range of knowledge and experience, and to apply them for better and faster development by supporting research proposals without limiting the disease area or the products categories.

LAT1 (L-type amino acid transporter 1: SLC7A5) is an amino acid transporter. While normal cells intake amino acids by LAT2, tumor cells intake amino acids by LAT1. In pancreatic cancer, LAT1 is overexpressed in tumor cells, and high expression of LAT1 is a predictive factor of poor prognosis. To date, LAT1 competitive-inhibitors, such as BCH and JPH203, were developed, and they are reported to be effective against various cancer cells in vitro and xenograft model. However, LAT1 non-competitive-inhibitors have not been developed.

Recently, we developed a LAT1 non-competitive-inhibitor and confirmed its anti-cancer effect against several cancer cells in vitro and xenograft model. Its oral administration also improved overall survival of genetically engineered mice with pancreatic ductal adenocarcinoma. After examining several nonclinical tests for the safety, we decided to move to the next step to acquire clinical proof of concept. To this end, we planned an investigator-initiated first in human clinical trial. After approved by institutional review board and clinical trial notification to PMDA, the trail has started and is ongoing. In the present session, we introduced our experienced process and various challenges to the investigator-initiated clinical trial.

It has been known that hepatocyte growth factor (HGF) is produced along with hepatic and renal disorders and promote regeneration of injured organs through the proliferative effects on (1) vascular endothelial cells, (2) myocardial cells, and (3) lymphatic endothelial cells. 

The mechanism of action of the HGF plasmid product is as follows; HGF plasmid is introduced by injection into skeletal muscle/myocardium, HGF protein is released outside muscle cells via the transcriptional /translational process within muscle cells, the angiogenesis and lymphangiogenesis though the mechanism of (1), (2), and (3) result in an improvement of symptoms of chronic obstructive arteriosclerosis, heart failure, and lymphoedema with amelioration of blood and lymphatic fluid circulation.

For the angiogenesis in the lower limbs, the data from the investigator-initiated clinical research at Osaka University (P1/2), company-sponsored clinical trials (P3), and the investigator-initiated clinical research by Advanced Medical Care B (P3) were used for the new drug application. For the angiogenesis in the heart, the investigator-initiated trial (P1) of Osaka University was conducted. For the lymphangiogenesis, the investigator-initiated trial (P2) of Asahikawa Medical University as well as the company-sponsored trial (P 1/2) are ongoing.

For development of products for treatment of rare diseases, as examples of industry-academia cooperation, we will introduce how we prepared for implementation of each investigator-initiated clinical research and investigator-initiated clinical trials, as well as what kinds of issues are there and how we have dealt with these issues to solve them.

In November 2016, we established the Life Intelligence Consortium (LINC), a collaboration organization between industry and academia and between life science and IT industries to advance AI development in the life science field. LINC consists of about 100 companies and organizations including IT companies and life science companies such as pharmaceuticals, medical care, and healthcare. With the support of academia from Kyoto University, RIKEN, etc., LINC aims to promote the health care field and related industries based on the AI strategy. In this lecture, I will introduce the details of research at LINC from the viewpoints of system pharmacology, ADMET, clinical data analysis, and natural language processing, and discuss the possibilities of AI and big data in the pharmaceutical industry.

Analysis of electronic medical record data, which is clinical real-world data, is expected to create new knowledge including new drug discovery targets. In LINC, projects such as stratification of diseases, detection of adverse events and discovery of unmet medical needs are in progress using electronic medical record data in collaboration with academia, life companies and IT companies. In this presentation, we will introduce an example of data analysis for renal disease using hospital-scale data conducted at Kyoto University Hospital, and discuss the possibility of using clinical real-world data for future drug discovery.

"Drug repositioning" has been attracting attention as a drug discovery strategy to overcome the recent slowdown in new drug development. Drug repositioning is a technique for discovering new effects of known drugs and redeveloping them as new indications for other diseases. Known drugs have already been confirmed for human safety and pharmacokinetics, and information such as compound production methods can be reused. Therefore, the drug repositioning approach is expected to reduce developed time, risk, and expenditure. Today, pharmaceutical companies are increasing to reuse their own drugs for the drug repositioning approach.

In Life Intelligence Consortium (LINC), pharmaceutical companies, IT companies, and academia are developing together in more than 30 projects. In this talk, we will introduce the developed AI technology in one of the projects, "drug repositioning". In this project, the AI models that predict the target protein, drug efficacy, phenotype, etc. from the compound structure is constructed. The developed AI models can be expected to contribute to the target prediction of active compounds with unknown mechanisms of action, search for new drug discovery targets, and the new indication prediction by reprofiling known drugs.

To efficiently drive drug discovery process, it is essential to improve the profile of ADMET, which is a collective term for endpoints related to pharmacokinetics and toxicity, as well as enhancing the activity and efficacy on a target protein. In this presentation, we will give an overview of the current status of ADMET prediction AI and de novo structure generation AI that are being promoted through AMED "drug discovery informatics project" and the activities of LINC (Life INtelligence Consortium), an AI drug discovery consortium on all Japan scale headed by Prof. Okuno in Kyoto University. Also, issues toward a new drug design AI system that combines above mentioned two types of AI models will be discussed.

The authors and G-Search Limited are collaborating to develop a search engine "JDream Expert Finder" (JDEF) as an outcome of the project-8 in the Life Intelligence Consortium (LINC). JDEF has a function to find young promising researchers, who, so far, have only a little publication in the literature database of Japan Science and Technology Agency (JST). Using conventional citation indicators such as h-Index, it is inadequate to measuring young researchers' performances. Therefore, we have focused on the betweenness centrality in co-authorship networks as an alternative indicator. As JDEF has utilized a growth model of Japan Society for the Promotion of Science (JSPS) research fellows, thus, it is able to search the other young promising researchers from the JST database.

Neurotrophins are one of the best-known examples of target-derived instructive cues that regulate neuronal development. Upon nerve growth factor (NGF) binds to its receptor TrkA at axon terminals, these complexes are internalized and retrogradely transported back to cell bodies. However, how neurons replenish TrkA in nerve terminals remains unknown. We show that retrograde signaling by NGF is necessary for soma-to-axon transcytosis of TrkA. Activated TrkA receptors are retrogradely transported to cell bodies, where they are inserted on soma surfaces and promote phosphorylation of resident naive receptors, resulting in their internalization. Prior to axonal transport, endocytosed TrkA is dephosphorylated by PTP1B to ensure targeting of inactive receptors to axons to engage with ligand. These results identify phospho-regulatory mechanisms of anterograde transcytosis of TrkA, which regulate neuronal sensitivity to NGF. We are now asking whether other membrane proteins are co-transcytosed with TrkA and we identify amyloid-beta precursor protein (APP) as a candidate. The TrkA-APP co-transcytosis regulates NGF functioning and APP metabolism. Since APP and its proteolytic products play an important role in pathogenesis of Alzheimer's disease (AD), NGF-dependent transcytosis might be also related to the onset of AD.

Pancreatic islets, the functional units in regulating blood glucose levels, are richly innervated by sympathetic nerves. Employing genetic mouse models and neuron-islet co-cultures, we discovered that sympathetic innervation is essential for organizing islet structure during development (Borden et al., Cell Rep. 2013). Further, we demonstrated that this inductive interaction at early stages is critical for mature islet function in regulating glucose metabolism, suggesting that developmental perturbations in sympathetic innervation might underlie metabolic dysfunction in humans. We are building on these findings to identify the molecular pathways by which sympathetic nerves instruct islet organization and the acquisition of functional maturity. To identify how sympathetic nerves influence islets, we recently asked whether neuronal activity is required for islet formation and function. Sympathetic nerves innervating the pancreas secrete the neurotransmitter, norepinephrine, which acts through pancreatic adrenergic receptors. We addressed the effects of blockade of sympathetic neurotransmission on islet morphology and function in mice via expression of tetanus toxin. Silencing of sympathetic activity resulted in defects in islet cyto-architecture and impaired expression of islet cell adhesion markers, similar to observations in sympathectomized mice. Surprisingly, "silenced" animals displayed improved glucose tolerance, enhanced glucose-stimulated insulin secretion, and higher insulin responsiveness, in contrast to findings in sympathectomized mice. These findings are consistent with previous studies describing an acute role for sympathetic activity in inhibiting islet insulin secretion and augmenting glucagon secretion. These results suggest that the presence of sympathetic nerves and neural activity exert similar effects on islet morphology during development but have distinct effects on islet function later in life.

Human mutations in the dystroglycan complex (DGC) result in not only muscular dystrophy, but also cognitive impairments. However, the molecular architecture critical for the synaptic organization of the DGC in neurons remains elusive. Here we report Inhibitory Synaptic protein 1 (InSyn1) is a critical component of the DGC whose loss alters the composition of the GABAergic synapses, excitatory/inhibitory balance in vitro and in vivo, and cognitive behavior. Association of InSyn1 with DGC subunits is required for InSyn1 synaptic localization. InSyn1 null neurons also show a significant reduction in DGC and GABA receptor distribution as well as abnormal neuronal network activity. Moreover, InSyn1 null mice exhibit elevated neuronal firing patterns in the hippocampus and deficits in fear conditioning memory. Our results support the dysregulation of the DGC at inhibitory synapses as a driver of altered neuronal network activity and specific cognitive tasks via a novel component, InSyn1.

Neuronal synapses are intimately ensheathed by abundant astroctytic perisynaptic processes, which is critical for synapse formation and function. In contrast to well-studied neuronal synaptic compartments, however, the molecular mechanisms of how astrocytic perisynaptic structures govern neuronal synapses remain ill-defined. Here, we develop a new in vivo chemico-genetic approach, Split-TurboID-GRAPHIC, that uses a cell surface fragment complementation strategy combined with informatics to identify the molecules at astrocyte-synapse junctions in vivo. We identify more than 100 proteins enriched at astrocyte-neuronal junctions. We find novel adhesion molecules highly expressed in cortical astrocytes whose deletion dramatically alters excitatory/inhibitory synaptic balance and also impairs spatial learning. Using Split-TurboID-GRAPHIC we thus establish a new mechanism by which astrocytes coordinate inhibitory synaptic balance with excitation via a chemo-affinity code of the tripartite synapse.

In 2019, the number of nursing universities increased to 272 schools in Japan. One out of three universities have a nursing school, and nursing universities continue to increase.

Ministry of Education, Culture, Sports, Science and Technology introduced the "Model Core Curriculum for Nursing Science Education in Japan" (MCCNSE) in 2017. MCCNSE aims at the acquirement of necessary and indispensable nursing competencies in the undergraduate course, which enumerates learning targets to be useful for making the curriculum.

MCCNSE has 7 areas to develop qualities and abilities of a nurse for a lifetime. A is basic qualities/abilities required of nursing professional. B is society and nursing science. C is basic knowledge necessary for understanding objects of nursing, includes pharmacological science. D is basic knowledge of specialty underlying nursing practice. E is basic knowledge necessary for nursing practice in various settings. F is clinical and regional training practice, and G is research of nursing science.

Nursing universities are required to comply with both the School Education Act and the Act on Public Health Nurses, Midwives, and Nurses. Nursing universities are expected to formulate the more complete and original curriculum through revision of Rules for Designation of Public Health Nurse, Midwife and Nurse Schools and Training Schools.

Most patients admitted to the hospital receive some form of medication. The nurse prepares the medicine, and checks as the final performer of pharmacotherapy, the "right patient" "right drug" "right purpose" "right dose" " right route" and the "right time", and give the patient medication. Furthermore, it confirms that the medicine is surely carried into the patient's body, observes the patient's reaction, and plays a role in quickly responding to any abnormalities. The medication by nurses is an extremely important and responsible task.

Will new nurses just graduate start working in hospital with the knowledge, skills, and judgment necessary to safely perform this important task? Their knowledge about drugs is fragmented, they do not integrate knowledge about diseases and conditions, drug actions and mechanisms, treatment guidelines and protocols. And they have not been acquired techniques of the medication intravenous injections, gastric catheter injection etc. They need to learn a lot after graduating from nurse school.

In the current acute ward, patients with different clinical conditions and treatment methods are admitted to the hospital, and the patient condition and drugs involved by nurses are diversified. It has become a difficult environment for nurses to safely carry out medication operations. Incidents related to nurse medications have not diminished, and nurses are concerned that they may become parties to the incident.

In hospital, we are exploring how to build work environments and educational systems for safety medication. In order for nurses to administer medicines that are safe for both patients and themselves, they must understand the dangerous environment around them and the knowledge and skills that are lacking in themselves. It is important that they can properly seek assistance from the surrounding staff.

In this symposium, I would like to think about what can be strengthened by basic education at school and what should be strengthened by on-the job education in order for nurses to administer medication safely.

Midwife is recognized as a responsible and accountable professional who contributes to the sexual and reproductive health/rights and welfare of individuals, families, and communities. In particular, midwife work in partnership with women to give the necessary support, care and advice during pregnancy, childbirth and postpartum period, to conduct normal births on her own responsibility with fully bringing out women's natural body functions, to support breastfeeding, and to provide care for newborns and infants. This care includes preventative measures, the promotion of normal birth and breastfeeding, the detection of complications in mothers and children, accessing of medical care or other appropriate assistance and the carrying out of emergency measures.

In light of this, midwife should firstly learn biological functions of women's body, as well as acquire knowledge on the in-vivo mechanisms of substances such as hormones, neurotransmitters and enzymes during pregnancy, childbirth, postpartum and breastfeeding period. Subsequently, midwife need to learn drug treatments to complement and support biological functions in case of disorders or impairments of women's body and mind.

In this revision of the midwifery curriculum, we intend to include the basics of drug treatment (pharmacological action, pharmacodynamics and drug interactions), individual drug treatments, and a wide range of knowledge from alternative drug treatments including herbal medicine, well as health food products and common food items. We hope to build a midwifery educational program on personalized, client-centered pharmacology to support mothers, newborns, infants, and childrearing families.

The 2017 model core curriculum and the 2019 proposed revisions to the designated rules call for the enhancement of pharmacology education necessary to exercise holistic assessment and clinical judgment skills.

The doctor prescribes according to the standard protocol considering the patient's condition and symptoms. The pharmacist confirms the details prescribed by the doctor and explains the efficacy and side effects of the drug to the patient or gives instruction on general precautions. The nurse's role is to watch the patient, check the effectiveness of the administered drug and adverse events, and report them to the doctor. To gain the expected efficacy from the drug and prevent an unwanted episode, however, the nurse should pay attention to the use of the drug.

In one case, the patient wearing a patch died after using an electric blanket, which caused a high blood concentration. Also, if a patient is dissatisfied with the prescribed medication and has difficulty maintaining medication adherence, the nurse needs to understand the reason why the patient has become unable to take medicine.

In other words, the nurse must be able to manage drug treatment and support medication after comprehensively understanding the patient's body, life, and psychological situation. To that end, the nurse needs to know the drug works, how to manage the treatment effect and safety, and how to use the drug on justified grounds. The author advocates the need for pharmacology education that integrates the knowledge of medicinal drugs and principles of nursing science so that the nurse can figure out the optimal medication method.

Statistical genetics is a research field that evaluates causality of human genetic variations on diseases, using statistical and bioinformatics approaches. Recent developments of sequencing technologies have provided human disease genome data of hundreds of thousands of the subjects, and successfully identified comprehensive catalogues of genetic susceptible loci. However, little is known regarding how to develop methodology to integrate large-scale human omics data with diverse biological resources, to which statistical genetics should contribute. We propose trans-layer omics analysis as a key to solve this challenging task. We have developed such methods and applied to a pioneering example of large-scale genetic association studies on a variety of human complex traits, including immune-related diseases. We demonstrated that the disease risk genes were significantly enriched in overlap with the target genes of the drugs currently used for treatment of the diseases, and that network analysis between the disease risk genes and the drug target genes could identify candidates of drug repositioning (e.g. CDK4/6 inhibitors for rheumatoid arthritis). These results should empirically show the value of statistical genetics to dissect disease biology and novel drug discovery.

Macrocyclic peptides possess a number of pharmacological characteristics distinct from other well-established therapeutic molecular classes, resulting in a versatile drug modality with a unique profile of advantages. Macrocyclic peptides are accessible by not only chemical synthesis but also ribosomal synthesis. Particularly, recent inventions of the genetic code reprogramming integrated with an in vitro display format, referred to as RaPID (Random non-standard Peptides Integrated Discovery) system, have enabled us to screen mass libraries (>1 trillion members) of non-standard peptides containing multiple non-proteinogenic amino acids, giving unique properties of peptides distinct from conventional peptides, e.g. greater proteolytic stability, higher affinity (low nM to sub nM dissociation constants similar to antibodies), and superior pharmacokinetics. The field is rapidly growing evidenced by increasing interests from industrial sectors, including small start-ups as well as mega-pharmas, toward drug development efforts on macrocyclic peptides, which has led to several de novo discovered peptides entering clinical trials. This lecture discusses the aforementioned screening technology, the RaPID system, and several showcases of therapeutic potentials of macrocyclic peptides.

Increasing evidence indicates that NLRP3 inflammasome plays a crucial role in the pathophysiology of cardiovascular diseases associated with sterile inflammation, including atherosclerosis and acute myocardial infarction (MI). NLRP3 inflammasome is a multimeric protein complex that leads to activation of caspase-1, which further induces maturation of interleukin (IL)-1β and IL-18. Activated caspase-1 also induces a particular form of cell death called pyroptosis via the cleavage of gasdermin D. We have shown that inhibition of the NLRP3 inflammasome attenuates the inflammatory response and ameliorates the severity of disease in murine models of cardiovascular diseases, such as atherosclerosis, MI, and aortic aneurysm. Moreover, the recent CANTOS trial showed that inhibition of IL-1β was efficacious in secondary prevention for cardiovascular events in patients with previous MI. These findings suggest that NLRP3 inflammasome may be a potential target for the prevention and therapy of cardiovascular disease. In this session, I summarize the current status of knowledge regarding the role of NLRP3 inflammasome in cardiovascular disease and discuss the prospects of NLRP3 inflammasome-targeted therapy.

In order to evaluate the emotional characteristics of mice, it is necessary to create an appropriate model and to adopt a useful behavior experiment device. The automatic hole-board test is an experimental method that can serve as a useful tool for evaluating the changes in various emotional states of mice. Especially, because the treatment with anxiolytics or anxiogenics and exposure to acute restraint stress affect head-dipping behavior, these behavioral changes may reflect the anxiogenic and/or anxiolytic state of mice. In addition, we created stress-adaptive and -maladaptive models in mice. A single exposure to restraint stress for 60 min produced a decrease in head-dipping behaviors of mice in the hole-board test, and these acute emotional responses were recovered by exposure to repeated restraint stress for 60 min/day for 7 or 14 days. However, mice that had been exposed to repeated restraint stress for 240 min/day for 7 or 14 days continued to show a decrease in head-dipping behavior in the hole-board test. Applying these models, we have recently found that prenatal stress exposure induces stress vulnerability and that histone deacetylase inhibitors can develop stress adaptation. In this symposium, we will introduce and discuss our recent findings.

Recently, various addiction problems have spread in the world, and the situation is growing serious. Addiction is a condition that results when individuals ingest an addictive substance or perform a specific action that can be pleasurable but the continuous use or act of which becomes compulsive and interferes with ordinary life responsibilities. It is very important to clarify the mechanisms underlying addiction, but there are still many unclear points. Animal studies have been crucial in understanding the biology and pathophysiology of drug addiction.

In recent years, although self-administration tests and conditioned place preference tests have been widely used as drug-dependence evaluation methods, intracranial self-stimulation (ICSS) method that electrically stimulate the medial forebrain bundle (lateral hypothalamic region) of the mesolimbic dopamine system could be also useful tools that can provide different insights in drug dependency assessment. In this symposium, we would like to discuss the advantage and disadvantage of ICSS method in drug dependence research and reward-system neural network analysis, in light of previous research and recent trends using the ICSS method.

We, MUROMACHI KIKAI, established in October 1959, are a manufacturer and distributor of animal research apparatus. In addition to our own products, we also sell imported products.

"Animal research apparatus" has a huge variety of types in terms evaluation, methods and etc.

To give some familiar examples, there are anxiolytic evaluation, antidepressant evaluation, and evaluation of antidementia, which is in other words learning and memory. Moreover, there are analgesia, dependence, motor function, regenerative medicine, and…, so there is no end.

A wide variety of evaluation exist. This means that there are considerable types of devices to do the evaluation, over the world. For example, some devices still carry out experimental methods that have been used for more than 30 years.

We have been involved in animal research instruments for a long time, and in the ever-advancing science technology, we are not only dealing with new products, but also continuing instruments that already have a long history, with some minor updates.

Looking back the highlights of our developments, there are several products that have been jointly developed according to the requests and wishes from researchers.

In this symposium, I would like to some examples of industry-academia collaboration in behavioral experiment apparatus such as automatic Hole-board test and Aggression response meter.

Spontaneous movement of the mouse is an important parameter that reflects the health and mental state of the mouse. In the present study, we tried to develop a new method that can measure the spontaneous movement of mice for 24 hours in a normal breeding environment using a simple and versatile video camera. Mice were housed in a standard cage in a 12 hour: 12 hour light / dark environment. The infrared lamp was used during the dark period. Mice were continuously photographed from above using a video camera. The position of the gravity center of the mouse in each frame of the captured video was calculated, and the amount of movement of the gravity center per second was expressed as the amount of exercise. We first confirmed that the momentum of mouse in the dark period was larger than that in the light period as is reported. When caffeine, a central nervous stimulant, was administered to mice, spontaneous motor activity increased until 3 hours after the administration, and then it returned to normal. When chlorpromazine, a sedative was administered to mice, the spontaneous movement of the mice almost disappeared, and the effect continued until 8 hours after the administration. In the present study, we succeeded in establishing a method that can analyze the spontaneous movement of mouse using a versatile video camera in a state close to a normal breeding environment. This system can analyze the spontaneous movement after drug administration.