正常個体中に存在する制御性T細胞 (Regulatory T cells, Treg)は、免疫自己寛容の維持、様々な免疫応答の抑制的制御に枢要である。内在性Tregの大部分は胸腺で、機能的に成熟した形で産生される。転写因子Foxp3は、Tregに特異的に発現し、Tregの発生、機能発現を制御するマスター制御遺伝子である。Foxp3+Tregの量的・質的異常は、様々な自己免疫／炎症性疾患の直接的原因となる。逆に、正常T細胞にFoxp3を発現させると、機能、表現型の点で内在性Tregと同等のTreg様T細胞に転換できる。一方、抗体、小分子を用いてTregの量的減少、抑制活性の減弱を図れば、がん免疫、微生物免疫を亢進できる。本講演では、Tregによる免疫抑制の分子機構、およびその機能、細胞系譜の維持機構について述べる。さらに、如何にTregを増やし、あるいは通常T細胞をTregに転換できるか、またTregを用いた細胞療法による自己免疫病、炎症性腸炎などの予防・治療が可能か、について議論する。

睡眠覚醒調節の根本的な原理、つまり「眠気」（睡眠圧）の脳内での実体とはいったい何なのか、またそもそもなぜ睡眠が必要なのか等、睡眠学の基本課題は全く明らかになっていない。私たちはこのブラックボックスの本質に迫るべく、ランダムな突然変異を誘発したマウスを8,000匹以上作成し、脳波測定により睡眠覚醒異常を示す少数のマウスを選別して原因遺伝子変異を同定するという探索的な研究を行なってきた。このフォワード・ジェネティクス研究の進展により、睡眠覚醒制御メカニズムの中核を担うと考えられる複数の遺伝子の同定に成功し、現在その機能解析を進めている。最近、フォワード・ジェネティクスによって同定されたSleepy変異マウスと断眠マウスの解析から、シナプス蛋白質の累積的リン酸化状態が睡眠圧の本態の一部である可能性が提示された。本講演では、筑波大学WPI-IIISの私どものラボにおける睡眠覚醒の謎への探索的アプローチを紹介する。

生体内では多様な輸送担体（トランスポーター;TP）が生体に必要な物質を積極的に取り込み、異物を積極的に排泄する機能を担っている。TPの分子認識に基づいた創薬を行うことにより、薬効を維持したまま副作用に関連する組織・細胞への移行を抑える理想的な動態特性を持つ薬の開発も可能になると期待され、医薬品開発の面からもTP研究への関心が高まっている。TPは、薬物代謝酵素と同様に、分子多様性、遺伝子多型、臓器特異性、広範な基質認識性という特性を持つことがわかってきた。医薬品開発においては、in vitroからin vivoへの外挿法などを確立していくことで、医薬品開発過程での遺伝子発現系の利用、薬物間相互作用の評価、遺伝子多型による個人間変動の解析などTP研究を実用段階へと進めていく必要がある。当日はその実例を示したいと思う。

近年、米国FDAが中心となり、生理学的薬物速度論モデル（PBPK model）による薬物動態予測を、臨床試験の必要性の判断、投与量の設定に活かそうという動きがある。この実例についても触れながらの講演にしたい。

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.

ミクログリアは中枢神経系での免疫担当細胞、起源として卵黄嚢の未分化マクロファージが源流であるとされている。正常時、ミクログリアは脊髄や脳に均等に存在し、小さな細胞体と長く可動性の突起をもち、それらを頻繁に動かして周辺の環境を監視しているように見える。病態時には、ミクログリアは活性化され、形態を変化させ、分裂増殖し、遺伝子発現の状況に変化を与え、例えばP2プリナージック受容体のような様々な神経伝達物質受容体の遺伝子発現に影響を与える。活性化ミクログリアはいくつかのATP 受容体サブタイプを発現する。外液のATPはこれらの受容体を刺激し、ミクログリアに様々な生理反応を引き起こす。その結果、例えば、P2X4受容体は神経障害性疼痛発症に重要な役目を果たす。また、P2Y12受容体およびP2Y6受容体は、それぞれミクログリアの遊走と貪食に重要な役割を持つ。この様な発見は、新たな医薬品創製の道を切り開くものである。実際に、P2X4受容体阻害剤は優れたFirst in classの神経障害性疼痛治療薬として有望視されている。医薬品開発には薬理学者が欠かせないと私は実感している。

ほ乳類の腸管には数百の腸内細菌（マイクロバイオータ）が存在し、宿主の生理機能に深く影響を及ぼしている。従ってマイクロバイオータに人為的に介入することが出来れば、複数の疾患に対する新たな治療戦略となり得る。我々は、消化管の恒常性維持機構を理解すると共に、個々の腸内細菌種が免疫システムにどのように影響を与えているかを還元化して把握して行く独自の研究手法を確立してきた。この方法によってこれまでに、制御性T細胞、Th17細胞、Th1細胞、CD8 T細胞を特異的に誘導する腸内細菌種の同定に成功した。これらの成果は、免疫系が関わる様々な疾患（感染症・アレルギー・自己免疫・がんなど）に対する新しい治療法開発に繋がる可能性がある。

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.

　シナプス可塑性の基礎研究については多くの知見が蓄積されている一方で、その臨床応用は非常に乏しいというのが現状である。グルタミン酸シナプスは脳内での情報処理において非常に重要な役割を果たしている。脳に可塑的な変化がおこる際、グルタミン酸受容体であるAMPA受容体がシナプスへ移行することは当研究室の研究も含めた多くの研究が明らかにしている。

　当教室ではAMPA受容体シナプス移行を促進する化合物を特定しており、脳卒中後のリハビリテーション効果促進薬としての可能性を模索している。現在げっ歯類においては劇的な効果が証明されており、霊長類においても同様の効果が見られている。今年度から臨床治験の開始を予定している。

　現在の精神神経疾患の診断治療は基礎研究の根拠に乏しい。本講演ではAMPA受容体を認識するPET Probeの開発について当教室の最新の知見を紹介する。AMPA受容体をヒトで可視化するPET Probeは未だ存在しない。この技術を用いて、「シナプス機能分子による精神神経疾患の再分類」が可能になり、今後の「基礎研究の根拠」に基づいた新規診断が可能になる。

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.

生体内には多様な脂質分子種が存在し、その質（リポクオリティ）の違いや代謝バランスの変化が,さまざまな炎症・代謝性疾患の背後に潜む重要な要素であることが示唆されている。一方で近年の質量分析技術の進歩は、質の異なる脂肪酸分子種の代謝、分布、動態を詳細に捉えることを可能にした。我々は、生体内の脂肪酸やリン脂質の代謝を網羅的かつ定量的に把握するためのリピドミクス解析システムを構築し、炎症・代謝性疾患の制御において脂肪酸代謝バランスが重要であることを示してきた。中でも、EPAやDHAなどω３脂肪酸が脂肪酸オキシゲナーゼにより活性代謝物に変換され、積極的に抗炎症作用や組織保護作用を発揮することを見出してきた。本講演では、生体内のリポクオリティが炎症制御や組織恒常性に及ぼす影響およびその分子機序について紹介する。さらに、生体内のリポクオリティの変化をノンバイアスかつ広範囲に捉えるための新技術、ノンターゲットリピドミクスの有用性と今後の発展性について議論したい。

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.

記憶形成時に引き起こされる可塑的刺激によって活性化された神経細胞集団において、シナプス入力が神経細胞の核内でCREB依存的転写誘導を引き起こす。その際の刺激依存的入力出力応答性を探索することにより、我々は、長期記憶・長期可塑性を制御するシナプスと核を結ぶ活動依存的神経可塑性メカニズムを明らかにした。シナプスから核、並びに、核からシナプスに至る情報伝達経路を解明することにより、複数のCREB経路と、その下流の標的遺伝子であるArcの制御に基づく逆シナプスタギング機構を発見し、これによりシナプス強度の強弱比が長期的に増強されることを見出した。一方、Arc転写可塑性とシナプス可塑性との間の両方向性機能連関のメカニズムを担うゲノム・蛋白の構造モジュールを活用することにより、世界に類のない、神経可塑性・神経活動のイメージングプローブE-SAREとXCaMPをも開発し、神経回路・脳情報動態研究を飛躍的に発展させる礎を築いた。本講演では、これらを応用した新たな光計測技術などによる脳機能制御メカニズム解明の例についても紹介したい。

既存の抗うつ薬の多くはモノアミンなどの神経伝達物質の作用を調節するが、一部の患者で十分な効果が得られないため、新たな創薬標的の開発が期待されている。近年、うつ病などの精神疾患患者の血中で炎症関連物質が高値を示すことから、炎症仮説が注目されるようになった。しかし、うつ病発症と炎症との因果関係は不明であった。社会や環境から受けるストレスは精神疾患のリスク因子であることから、動物に繰り返しストレスを与える反復社会挫折ストレスがうつ病の動物モデルとして使用されている。反復ストレスは情動変容を誘導し、同時に特定の脳領域のミクログリアを活性化することを見出した。この研究に端を発し、反復ストレスによる情動変容の誘導における脳内炎症の関与とその分子実体を明らかにした。また、脳内炎症の関与がすでに知られている神経変性疾患で、炎症を標的とした創薬基盤を開発した。本講演では、精神・神経疾患の病態形成に関与する脳内炎症について最新の知見を紹介したい。

うつ病は、患者個人のみならず社会経済に与える損失が大きい身近な精神疾患であり、全世界のうつ病患者数は2億6400万人以上と推計されている(WHO, 2019年)。モノアミン神経系に作用する既存の抗うつ薬は、効果発現までに数週間以上を要し、約3割の患者には無効である(治療抵抗性うつ病)。一方、近年の研究により、NMDA受容体拮抗薬ケタミンが、治療抵抗性うつ病患者に即効性の抗うつ作用を示すことが明らかにされているが、依存性や統合失調症様症状の惹起などの重大な副作用のため、ケタミン自体の抗うつ薬としての使用には大きな制約がある。そのため、ケタミンより副作用の少ない新たな即効性抗うつ薬の開発が求められている。私は、ドコサヘキサエン酸およびエイコサペンタエン酸の活性代謝物レゾルビン類に着目し、レゾルビン類がケタミンと類似のメカニズム(mTORC1経路活性化)で抗うつ作用を示すことを発見した。レゾルビン類は、恒常性維持に関与する内因性脂質メディエーターであることから、ケタミンより安全性面で優れた新たな即効性抗うつ薬の創薬標的になると期待できる。

プロスタグランジンD₂(PGD₂)は、マスト細胞が起点となるアレルギー性炎症において大量に分泌されるPGである。PGD₂は長らくアレルギー性炎症を促進すると考えられてきたが、抑制作用を示した報告もあり、その役割には不明な点が多く残っていた。講演者らは、全身性およびマスト細胞特異的PGD合成酵素欠損マウスを用いた検討から、PGD₂がその受容体であるDP1を介して食物アレルギーやアナフィラキシーの進行を抑制することを明らかにした。さらに、DP1受容体作動薬がそれらの病態進行を強く抑制することも見出した。現在は、もう一つの受容体であるDP2の役割解明やPGD₂の尿中代謝産物を用いた食物アレルギーの新規重症度診断法の開発をすすめている。本講演では、これまで明らかにしたPGD₂の役割とその応用可能性を紹介したい。

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.