National Children's Medical Research Center

HISTORY
 The National Children's Medical Research Center (NCMRC) was established in October 1984 as an adjunct medical research institute affiliated with National Children's Hospital. The National Children's Hospital is a sole medical care center for the children in this country under the auspices of the Ministry of Health and Welfare.
 At its inception, four Departments and eight laboratories were founded, i.e., Department of Pathology and Pathophysiology, Department of Endocrinology and Metabolism, Department of Congenital Abnormalities Research, and Department of Pediatric Ecology. At the next year, in October 1985, two Departments and one Laboratory were established in the Institute. They were Department of Immunology and Allergy, Department of Infectious Disease, and Multidisciplinary Laboratories.
 Furthermore, Department of Experimental Surgery and Bioengineering in October 1986, and Department of Pediatric Pharmacology in October 1987 were established. Thus, till to now the Institute bears 8 departments and 16 laboratories.
 In March 1996, Genox Research Institute focusing on development of new medicine targeting allergic disease related novel gene, was established as an annex to NCMRC.

 INTRODUCTION
 The primary mission of NCMRC is to conduct and coordinate research projects in basic and applied medical sciences aimed at the understanding of the causes of children's diseases, and also developing new and better means of prophylaxis, diagnosis and treatments. The affiliation with National Children's Hospital facilitates clinically oriented research projects, as well as those in basic medical sciences, thereby providing ideal research environments. The research activities of 32 official faculty members and their associates encompass a wide range of molecular biology in basic and clinical medical sciences, biomedical engineering, and psychology reflecting recent development in technology and also the present and future needs for the advanced care of sick children. In the past much efforts have been devote to reduce an annual death rate of newborn children, and hence Japan is now considered to be one of the most advanced countries in the world in terms of the least rate of newborn child death. Nevertheless, a cause death of the newborn children with congenital abnormalities ranked as one of the highest and showed nearly 35 % of the death. Therefore, based on the developmental aspects of the disease, more synthetic research and clinical cares are required. Thus, we are focusing on an overall goal to solve the various aspects of child development and its abnormalities. Our research fields thus include molecular and cellular biology, developmental biology, immunology, pharmacology, as well as social and medical sciences.
 Current acting members who engage in each research project are 32, and the research projects of the center are funded by the Department of Health and Welfare, Health Sciences Research Grant, and other governmental funding agencies. It also includes many other foundations as well as foreign agencies. A current annual budget of research grant is reaching approximately 1,000,000,000 yen per year including core grant of Organized Research Combination System with Encouragement of Center of Excellence (COE) and NCMRC based Contract Research Grant funded by the Department of Health and Welfare.
 Inasmuch as the current social issues of having fewer children are serious problems for future medical cares, a more synthetic medical study including children as well as maternity and paternity is required. As a leading medical center to cover a border and to solve related medical issues, the center will be reborn as The National Medical Center for Developmental Health Care with advanced and high technology in the year of 2001 after combined and reorganized by the National Children's Hospital and Ohkura Hospital. (September 1 1999)

ORGANIZATION
    Department of Pathology and Pathophysiology
        Laboratory of Pathology
        Laboratory of Pathophysiology
    Department of Endocrinology and Metabolism
        Laboratory of Endocrinology
        Laboratory of Metabolic Disorders
    Department of Congenital Abnormalities Research
        Laboratory of Genetics and Cytogenetics
        Laboratory of Teratology
    Department of Child Ecology
        Laboratory of Environmental Epidemiology
        Laboratory of Interactive Development
    Department of Allergy Research
        Laboratory of Immunology
        Laboratory of Allergy
    Department of Infectious Diseases Research
        Laboratory of Bacteriology
        Laboratory of Virology
    Department of Experimental Surgery and Bioengineering
        Laboratory of Experimental Surgery
        Laboratory of Bioengineering
    Department of Pediatric Pharmacology
        Laboratory of Toxicology
    Division of Research Promotion
    Genox Research, Inc.


Department of Pathology and Pathophysiology

Laboratory of Pathology

 Main research subject is to analyze the mechanism of growth and differentiation of blood cells and their malignant counterparts, namely leukemias. Especially, foci are directed toward the signal transduction of B-precursor cells and their malignancies, the mechanism of DNA polyploidization in megakaryocyte maturation, and the effect of granulocyte colony-stimulating factor in immune system. Analysis of organ dysfunction caused by verotoxin producing bacteria is also conducted using renal tubular and endothelial cells.
Projects
1. Molecular basis of hematopoiesis and its dysregulation
  1) Function of granulocyte colony-stimulating factor
  2) Megakaryocyte differentiation and platelet production
  3) Tyrosine phosphorylation in early B-cells
  4) Tyrosine phosphorylation in allergy-related cells
  5) Diagnosis of childhood hematopoietic malignancies
2. Mechanism of organ failure induced by verotoxin (Shigatoxin)
  1) Distribution of verotoxin binding sites in human kidney
  2) Mechanism of renal tubular cell apoptosis
  3) Mechanism of Burkitt's lymphoma cell apoptosis
  4) Cytotoxic activity of verotoxin toward endothelial cells
  5) Development of assay for early detection of renal failure
3. Molecular mechanism of cell cycle regulation

Laboratory of Pathophysiology

 Research from this laboratory, directly related to the treatment of severely ill pediatric patients, has produced results which are actually being put to use today. The complicated nature of circulatory and respiratory physiology and pathology is being analyzed by using the methods of applied physiology, without being limited to the molecular or cellular microscopic level. The results are being put to use for the development of artificial ventilators, devices to measure cardiopulmonary function, artificial organs and even more effective methods of treatment involving telemedicine.
Projects
Pathophysiology and treatment of critically ill children
1. Optimal use of high frequency oscillatory ventilation
2. Etiology and strategy for lung injury prevention
3. Safe application of nitric oxide inhalation therapy
4. Heparin coated extra corporeal membrane oxygenation (ECMO)
5. Physiological effect of endoscopic surgery in children
6. Hepatorenal effect of volatile inhalational anesthetic agent, sevoflurane
7. Prevention of sudden infant death syndrome
8. Near infrared spectroscopy as an intracranial oxygen and hemodynamic monitor in children
9. Development of pain management devices in pediatric patients
10 . Health telematics and home care pediatric patients
11. Organ transplantation in children and optimal method of transport


Department of Endocrinology and Metabolism

Laboratory of Endocrinology

 Main research projects of Endocrine Research Laboratory are growth, which is the characteristics in children, and sexual differentiation and maturation. We study on clinical oriented projects, such as mechanism of growth, diagnosis and treatment of growth hormone deficiency, establishment of treatment for non-growth hormone deficient short children, classification and diagnosis of disorders of sex differentiation, treatment of precocious puberty or hypogonadism. Molecular approach to endocrine and metabolic disorders such as congenital adrenal hyperplasia, testotoxicosis, GH deficiency Type 1A, Kallmann syndrome is investigated in cooperation with other institutes.
Projects
1. Growth mechanism
  1) Standardization of free IGF-1 and ALS
  2) Study on 20K hGH
  3) Computer-aided automatic measurement of bone age
2. Clinical and basic approach to children with growth disturbance
  1) Prediction of final height in short children
  2) Bone mineral density in endocrine diseases
  3) DNA analysis of GH or GH receptor abnormality
  4) DNA analysis in short stature
  5) High dose GH treatment in children with intrauterine growth retardation
  6) Psychological approach to short children
  7) Treatment in children who enter puberty at short stature
3. GH deficiency
  1) Frequency of GH deficient children
  2) Standardization of value measured by GH kits
  3) Biological assay for hGH
4. Clinical and basic study in obese children
  1) Growth mechanism in obese children
  2) Obese children and leptin
5. Study on sexual differentiation and sexual maturity
  1) Treatment in children with central precocious puberty with super-long acting analog
  2) Social sex in children with abnormal sexual differentiation
  3) hCG-hMG treatment in male hypogonadism
6. Molecular approach to endocrine diseases
  1) Study on GHF receptor
  2) Cloning of vitamin D hydroxylase and clinical characteristics of vitamin D resistant richets

Laboratory of Metabolic Disorders

 Study on physiological role of biotinidase, which metabolites biotin is the main project of Metabolic Research Laboratory.
Projects
1. Study on physiological roles of biotin and lipoic acid
  1) Study on effects of biotin on membrane-bound enzymes and/or receptors
  2) Study on effects of biotin supply on diabetes mellitus
  3) Development of a method to determine lipoic acid
2. Study on biotinidase and lipoamidase
  1) Isolation and chemical characterization of human serum biotinidase
  2) Estimation of the origin (s) of serum biotinidase
  3) Isolation and chemical characterization of human urine biotinidase
  4) Isolation and chemical characterization of human breast-milk biotinidase
  5) Synthesis of novel substrates for biotinidase and lipoamidase
  6) Study on enzyme kinetics of biotinidase and lipoamidase
  7) Study on roles of glycosylation of biotinidase and lipoamidase
  8) Study on biotinidase and lipoamidase in atopic dermatitis
  9) Study on biotinidase and lipoamidase in diabetes mellitus
3. Study on molecular basis of endocrine and metabolic diseases
  1) Study on molecular basis of congenital adrenal hypoplasia
  2) Study on molecular basis of congenital unresponsiveness to adrenocorticotropin
  3) Study on molecular basis of congenital adrenal hyperplasia
  4) Study on molecular basis of apparent mineral corticoid excess
  5) Study on molecular basis of bone diseases
  6)Study on molecular basis of familial male-limited gonadotropin independent precocious puberty
  7) Study on molecular basis of McCune-Albright syndrome
  8) Study on molecular basis of isolated growth hormone deficiency


Department of Congenital Abnormalities Research
Laboratory of Genetics and Cytogenetics, and Laboratory of Teratology

 We are studying on the structure and function of the human genes using molecular and biochemical techniques to elucidate the molecular mechanism in pathogenesis of hereditary diseases as well as childhood tumors, and to apply the information to prevention and therapy. Our researches include determination of the chromosomal location of an unidentified disease gene (mapping), identification of responsible genes by positional cloning strategies and/or by candidate gene approaches, and detection of mutations in patients' genome. Our Department is proud to be a pioneer to identify causative mutations and genes for three disorders: the amelogenin gene (AMG) for X-linked amelogenesis imperfecta, trinucleotide repeat expansion for dentatorubral-pallidoluysian atrophy (DRPLA), and a unique missense mutation of PAX6 for isolated foveal hypoplasia.
 Currently, our efforts to identify the gene and novel mutations focus on a various types of congenital eye anomaly, renal insufficiency and infantile jaundice. We are also analyzing molecular mechanisms for the transcriptional control by WT1 and PAX6, and for apoptosis in leukemia cells treated with chemotherapeutants as well as in cells expressing expanded triplet repeats. We are also conducting basic researches with cultured cells and model animals for gene therapy.
Projects
1. Molecular studies on genetic diseases and its application to clinical medicine.
1) Identification of genes responsible for and involved in genetic diseases and establishment of the genotype phenotype correlation.
2) DNA diagnosis for genetic diseases and improvement of its techniques.
3) DNA polymorphisms and mapping.
4) Human genome project.
5) Studies on molecular pathogenesis of triplet repeat disorders including dentatorubral- pallidoluysian atrophy (DRPLA).
2. Molecular studies on oncogenes and tumor suppressor genes involved in childhood tumors.
1) Identification of genes responsible for and involved in childhood tumors.
2) Functional analyses of the WT1 gene responsible for Wilm's tumor.
3. Molecular studies on expression regulation in mammalian cells.
1) Expression regulation of disease genes.
2) Expression regulation of pituitary hormone genes.
4. Molecular studies on apoptosis and its application to clinical medicine.
1) Apoptosis in the onset and remission of leukemia.
2) Apoptosis induced by extended polyglutamine tract.
5. Gene therapy
1) Vector systems introducing exogenous genes to hepatocytes.
2) Survival of allograft transplanted organs by suppression of the immune reaction by the use of genes involved in apoptotic process.
3) Suppression of apoptosis in hepatocytes with the Bcl-2 expression.


Department of Child Ecology

Laboratory of Environmental Epidemiology

 This division is aiming at investigation of etiology and prevention of childhood disorders. The main tool of research is epidemiologic approach with the use of computer. The major concern and ongoing research topics include: multi-center clinical trial of childhood leukemia in which a new treatment protocol is evaluated and new risk factors are sought for; investigation of risk factors of childhood malignancy based on the tumor registry of the National Children's Hospital; quantitative evaluation of family history in the prevention of atherosclerotic disease from childhood; long-term follow-up of low-birth-weight babies and evaluation of their physical and psychological status; growth hormone administration and the relapse of brain tumors and the development of leukemia; electromagneic waves and childhood leukemia; genetic epidemiology of prenatal risk factors and of growth and development of children.
Projects
1. Epidemiology of Childhood Diseases
  1) Multi-center clinical trial of acute lymphoblastic leukemia
  2) Quantitative assessment of family history of atherosclerotic diseases
  3) Survival and risk factors of childhood malignancy based on the inpatient registry
  4) The role of aldose reductase in complications of diabetes mellitus
  5) Growth hormone administration and recurrence of brain tumors
2. Genetic epidemiology
  1) Genetic epidemiology of prenatal risk factors, growth and development
3. Health promotion, psychology and other studies for children
  1) Prevention of atherosclerotic diseases from childhood
  2) Psychological and social assessment of short-statued children
  3) Establishment of reference values for pediatric laboratory tests

Laboratory of Interactive Development

 Understanding the disorders and the diseases of the developing children in the light of the host-environmental interaction is our department's priority. Our special interest is focused on the development of mucosal immunity of the infants and on the epidemiological study of the various intractable diseases in a childhood. Using molecular and cell-biological techniques as well as epidemiological analysis, the following studies are in progress.
Projects
1. Nation-wide survey of childhood cancer and chronic diseases
  1) The Japan children's cancer registry
  2) Genetical and epidemiological study on childhood cancer and chronic diseases
  3) The child abuse survey in pediatric clinics in Japan
  4) The registry for congenital diseases in urban areas
2. Development and disorders of mucosal immunity in children
  1) Role of the intestinal microflora in mucosal immunity and oral tolerance
  2) Pathogenic mechanism of the infection by the enterohemorrhagic Escherichia coli
  3) Influence of the proton pomp inhibitors on mucosal immunity
3. Role of the host-environmental interactions in patients with intractable diseases
  1) Genetic factors that predispose Japanese population to Kawasaki disease
  2) Search for a pathogenic microbe in patients with Kawasaki disease
4. Child health and development
  1) Quality of life for chronically-ill children
  2) Effects of life environment on child health and development
  3) Effects of sociocultural factors on child development


Department of Allergy Research

Laboratory of Immunology

 The refractory inflammatory diseases in children are investigated here by various immunological methods. The main projects include researches on roles of costimulatory molecules in the carcinogenenesis, GVHD and autoimmune diseases, and how T cell activation by superantigen influences pathodology of vasculitis. The bile duct diseases such as biliary atresia and Allagile syndrome, are also studied from the genomic (Jagged 1 gene) and viral (HCMV) aspects.
Projects
1. Analysis of lymphocyte functional molecules in the immunological tolerance and the development of immune diseases.
  1) Functional analysis for costimulatory molecules involved in the development of immune diseases.
  2) Development of a new therapeutic approch for immuneintervention by manipulating costimulatory molecules.
  3) Mechanisms of signal transduction through T cell costimulatory molecules such as CD28 and CTLA-4.
2. Immunological analysis of the vasculitis induced by superantigen or verotoxins.
  1) Analysis for the mechanism of brain and renal disturbance.
  2) The role of YPM protein deribed from Yersinia pseudotubercrosis in the development of vasculitis.
3. Analysis for pathogenesis of the bile duct and hepatic diseases.
  1) Abnormalities of Jagged 1 gene and the pathogenesis of the bile duct and hepatic diseases.
  2) The correlation of cytomegarovirus infection and the development of bile duct stresia.

Laboratory of Allergy

 Type I allergy is promoted by binding of IgE antibodies that are produced by IL-4 stimulation to mast cells. The functional roles of IL-4 and mast cells are recently found to vary depending on species, i.e., between mouse and human. We have established the method of generating a large amount of human mast cells in culture. Therefore, we are mainly investigating the functional and genetic changes in activated human mast cells and other inflammatory cells.
Projects
1. Studies on cells involved in allergic inflammation.
  1) Development of mast cells, eosinophils and basophils.
  2) Functional analyses of human mast cells, basophils and eosinophils.
  3) Cytokine production by epithelial cells and fibroblasts.
2. Studies on environmental and food allergens.
  1) Studies on environmental allergens.
  2) Studies on food allergy.
3. Basic studies on atopic dermatitis.
  1) Staphylococcal antigens.
  2) Molecular analyses of intractable atopic dermatitis.
4. Genes expressed in relation to pathological changes in atopic diseases.


Department of Infectious Diseases Research

 Pathogenesis and host defense mechanisms of intractable infectious diseases of children
 Aims to clarify the molecular mechanisms of bacterial infections toward the development of rapid diagnosis, prevention and specific therapy. Current subjects are as follows: 1) Enterohemorrhagic Escherichia coli O157:H7 infection including hemolytic uremic syndrome. 2) Superantigen-mediated diseases such as Yersinia pseudotuberculosis infection including Kawasaki Disease.
 Host Defense Research Laboratory
 Aims to clarify the defense mechanisms against infection. Current subject is the molecular mechanisms of superoxide anion produced by phagocytes. Chronic granulomatous disease is one of the targeted illnesses.
Projects
1. Studies on the enterohemorrhagic Escherichia coli O157 infection
  1) Molecular mechanisms of the hemolytic uremic syndrome (HUS)
  2) Rapid diagnosis and specific treatment of the enterohemorrhagic E. coli infec-tion.
  (1) Clinical study of the Shiga toxin-absorbing drug (Synsorb Pk)
  (2) Development of a humanized anti-Shiga toxin antibodies
  (3) Development of rapid and simple detection technique of fecal O157 antigen and Shiga toxins
2. Host-defense mechanisms by phagocytes
  1) Studies on the NADPH oxidase system responsible for anti-infectious activity of phagocytes
  2) Structural and functional analysis of the defective NADPH oxidase in chronic granulomatous disease (CGD)

Laboratory of Bacteriology

 We have been working on the non-invasive prenatal diagnosis using fetus' blood obtained in the peripheral blood of the pregnant woman.
 We have also been working on the identification of the microbial species and toxic genes.
Projects
1. Non-invasive prenatal diagnosis using fetus' blood in the peripheral blood of pregnant woman
2. Identification of microbial species and toxic genes

Laboratory of Virology

 Our major interest focuses on molecular mechanisms of childhood malignancies. In the case of childhood malignancies, recent evidence suggests genetic background for each patient plays important roles for disease susceptibility. Thus studies on the genetic control of DNA repair, cell cycle and apoptosis in response to environmental stresses are being conducted in our department.
Projects
1. Cell cycle and apoptosis regulation in tumor development and cell differentiation.
  1) Elucidation of novel a function of CDK inhibitor p21 depending on its subcellular localization.
  2) Cloning and functional analysis of a novel molecule Kim-1, which is apparently involved in cell cycle regulation.
2. Molecular mechanisms for cancer development in cancer prone families.
  1) Cancer susceptibility and cell cycle and apoptosis dysregulation in Ataxia telangiectasia.
3. International collaborative work in molecular epidemiology and genetic back-ground of infantile leukemia.


Department of Experimental Surgery and Bioengineering

 For the radical therapy in pediatric patients with severe organ failure, we have been investigating the organ transplantation and the reconstruction of organ tissues in the field of experimental surgery, as well as the biomolecular immunoregulation after grafting and the development of substitute organs in the bioengineering field.

Laboratory of Experimental Surgery

 For establishment of organ transplantation as a radical therapy for congenital and acquired organ failure, following studies are ongoing in this division: induction of recipient immunosuppression, analysis of transplantation tolerance and microchimerism, finding of new genes related to tolerance induction, and reconstruction of hepatocytes for the severe liver failure.
Projects
1. Study for immunoregulation in the organ transplantation
  1) Immunoregulation by inducing lymphocyte apoptosis and study of its biological mechanisms
  2) Preclinical study of a novel immunosuppressant FTY720 using canine model of kidney transplantation
  3) Prevention of graft rejection and induction of transplantation tolerance by pretransplant donor treatment
3. Fundamental study for the relationship between transplantation tolerance and microchimerism following organ transplantation
4. Investigation of the genes related to transplantation tolerance
5. Study for reconstruction of the liver in patients with hepatic failure at an end-stage

Laboratory of Bioengineering

 The current aims in this division are to perform the basic research for induction of immunosuppressive genes into grafted organs, gene therapy and cell transplantation for metabolic organ disease, and development of hybrid artificial liver.
Projects
1. Immunoreguration after organ transplantation and radical treatment of metabolic disease by the therapy using gene-introduction
2. Study for developing an artificial hybrid liver


Department of Pediatric Pharmacology
Laboratory of Toxicology

 The Human Genome Project now hurtles toward completion of the first finished human sequence (now scheduled for 2003). In the near future, therefore, interest for biology would be predicted to be shifted from structure of genome to function (Functional Genomics). Hence, we view that future biological research should be based on the Genomic Information, and our Department is focusing on studying the molecular mechanism of a variety of multi-gene disorders and innovating novel therapeutics for them. More specifically, we are now establishing a novel approach for Drug Discovery based on the Genome information by using G-protein coupled receptor as a model. "Dry Biology" (bioinformatics) and "Wet Biology" (molecular biology and cell biology) would be integrated for the purpose.
Projects
  1) Study on developmental alteration in expression and function of G-protein coupled receptors
  2) Molecular mechanism for change in disease states
  3) Structure-activity relationship of drugs that act on G-protein coupled receptors  4) Signal transduction, subcellular localization, physiological role and drug action of G-protein coupled receptors
  5) Drug discovery by using cloned G-protein coupled receptors
  6) Assessment of physiological role of G-protein coupled receptors signaling by using gene-target animals


Division of Research Promotion

Research Administrations :
 For the purpose of promoting research activities in the National Children's Medical Research Center (NCMRC), the Department of Research Promotion is corresponding to the administrations of common facilities, equipments and research cooperations. In terms of common facilities and equipments, the NCMRC has the Radioisotope Center, the Experimental Animal Center, the Gene Engineering Center and common equipments. These systems are usually under the control of individual committees composed of NCMRC's stuffs. The promotion of research cooperations among institutes and/or National Children's Hospital (NCH) is also performed by the Research Planning Committee and the Joint Research Committee for NCH and NCMRC.
Research Activities:
Following researches are now undergoing.
 The study of pharmacological action mechanisms of a novel immunosuppressant FTY720.
 The study of apoptosis inducing agents and cancer research.
 The research of chemical substance safety tests for new born and fetal animals.
Projects
1. Research in control of immunological responses
Study of mechanisms on the bases of apoptotic induction in lymphocytes
2. Research in infantile carcinoma
Study of anti-cancer effect of apoptosis inducing agents
3. Research in developmental toxicities of chemical substances
Establishment of evaluation methods of toxic substances by means of rat hepatocytes primary cultures


Genox Research, Inc.

 The mission of Genox Research Inc. is to identify novel therapeutic targets for allergic diseases such as rhinitis, bronchial asthma and atopic dermatitis in close collaboration with National Children's Hospital.
 We are studying T cells, eosinophils, and monocytes isolated from peripheral blood of allergy patients. Genes expressed in those cells are displayed by fluorescent differential display technology. Comparison of the display patterns of the patient and control samples reveals the genes preferentially up- or downregulated in patients. The level of expression of these genes is also examined by quantitative PCR to determine the relevance of a particular gene to allergy. Using these procedures, several candidate genes have been identified thus far, and functional analysis of these genes is now in progress.
 Genox is also developing the Database which integrates data on gene expression and the clinical features of allergy patients. This database will be helpful in understanding the cross talk between genes in the pathogenesis of allergy and in identifying new targets for drug discovery.
Project
Project 1 Preparation and banking of research materials from clinical samples
  1) Isolation of peripheral blood leukocytes from allergy patients and immortaliza-tion of B lymphocytes
  2) Preparation of RNA from human leukocyte samples
  3) Preparation of cDNA libraries from human leukocyte samples

Project 2 Allergic disease related gene expression analysis
  1) Gene expression analysis by differential display methods
  2) Sequencing of the expressed genes
  3) cDNA cloning
  4) Gene expression analysis by real time quantitative RT-PCR and DNA microarray
  5) Screening of associated genes by yeast two hybrid systems
  6) Analysis of gene function by immunobiological studies
  7) Functional study using in vitro-differentiated mast cells and T cells

Project 3 Bioinformatics
  1) Development of Genox database managing materials and clinical and experi-mental information
  2) Development of data mining systems for gene expression analysis by linking experimental data and external data with clinical information