<clinical_study>
  <!-- This xml conforms to an XML Schema at:
    https://clinicaltrials.gov/ct2/html/images/info/public.xsd -->
  <required_header>
    <download_date>ClinicalTrials.gov processed this data on September 20, 2023</download_date>
    <link_text>Link to the current ClinicalTrials.gov record.</link_text>
    <url>https://clinicaltrials.gov/ct2/show/NCT02217384</url>
  </required_header>
  <id_info>
    <org_study_id>S-20140040</org_study_id>
    <nct_id>NCT02217384</nct_id>
  </id_info>
  <brief_title>Whole Gut Microbiome Sequencing in Patients With Anorexia Nervosa</brief_title>
  <official_title>Whole Gut Microbiome Sequencing in Patients With Anorexia Nervosa</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>Odense University Hospital</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>University of Copenhagen</agency>
      <agency_class>Other</agency_class>
    </collaborator>
    <collaborator>
      <agency>Aarhus University Hospital</agency>
      <agency_class>Other</agency_class>
    </collaborator>
    <collaborator>
      <agency>Aalborg University Hospital</agency>
      <agency_class>Other</agency_class>
    </collaborator>
    <collaborator>
      <agency>INRA Research Center at Jouy-en-Josas</agency>
      <agency_class>Other</agency_class>
    </collaborator>
  </sponsors>
  <source>Odense University Hospital</source>
  <oversight_info>
    <has_dmc>Yes</has_dmc>
  </oversight_info>
  <brief_summary>
    <textblock>
      Background. Anorexia nervosa (AN) still carries the highest fatality rate of any psychiatric&#xD;
      disease, and less than half of the patients recover, completely refractory to any treatment.&#xD;
      The etiology remains unknown and evidence for treatment is lacking. The intestinal microbiota&#xD;
      and its microbiome provide humans with additional gene products which may be regarded as an&#xD;
      organ, which contributes to multiple host metabolic pathways. Recent advances in microbial&#xD;
      DNA sequencing technologies have resulted in metagenomic DNA analysis of whole ecosystems&#xD;
      such as the human gut. Altered intestinal microbiota has been related to obesity and insulin&#xD;
      resistance. Hypothetically, the intestinal microbiota could play a role in the generation&#xD;
      and/or maintenance of the emaciation in AN.&#xD;
&#xD;
      Aim. The aim of the present study is to investigate whether gut microbiota is altered in&#xD;
      patients with AN.&#xD;
&#xD;
      Subjects and methods. A cross sectional study of the gut microbiome profiles in 75 clinical,&#xD;
      psychometric and biochemical well characterized treatment seeking females with AN. The&#xD;
      microbiome profiles are compared with 75 age- matched healthy Danish control subjects.&#xD;
&#xD;
      Perspectives. Clarifying whether the intestinal flora is implicated in the susceptibility to&#xD;
      or maintenance of AN may provide the basis for development of new highly required treatments.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      BACKGROUND Anorexia nervosa (AN) is a complex disorder characterized by disturbed body image,&#xD;
      ego-syntonic neglect, ambivalence, self-starvation, loss of body weight, obsessive thoughts&#xD;
      of food, ritualistic patterns of food intake, elevated physical activity, depression, anxiety&#xD;
      and emotional rigidity. The evidence base for the treatment is very limited, considering the&#xD;
      extent to which this disorder erodes quality of life and still carries the highest fatality&#xD;
      rate of any psychiatric diseases. In fact, there is no evidence that the prognosis has&#xD;
      improved throughout the 20th century.&#xD;
&#xD;
      AN remains a syndrome, i.e. collections of symptoms, as it is not defined by etiology. Twin&#xD;
      studies have consistently demonstrated that AN is strongly familial predominantly due to&#xD;
      genetic factors. However, neither candidate gene nor genome-wide studies have identified&#xD;
      truly validated genes for AN. AN is associated with multiple, profound endocrine&#xD;
      disturbances. However, so far the extensive research has not led to any breakthrough&#xD;
      discovery. There is a need for new thinking.&#xD;
&#xD;
      Colonization of the human gut begins at birth with bacteria from the mother's vagina and&#xD;
      labor-released maternal feces. In adults the human digestive tract contains complex&#xD;
      assemblies of microorganisms which actually outnumber the number of the host cells 10:1. This&#xD;
      intestinal microbiota and its microbiome provide humans with additional gene products, which&#xD;
      may be regarded as an organ, which contributes to multiple host metabolic pathways. Thus, a&#xD;
      whole range of substances are produced which are not yet identified. Potentially some of&#xD;
      these substances may affect functions in the brain, such as appetite and emotion regulations.&#xD;
      Recent advances in microbial DNA sequencing technologies have resulted in metagenomics DNA&#xD;
      analysis of whole ecosystems such as the human gut. An important step forward in&#xD;
      investigating the bacterial composition of the human microbiome was taken with the Metagenome&#xD;
      of the Human Intestinal Tract consortium reporting the first complete microbial gene atlas of&#xD;
      the human distal gut, using deep next generation metagenome sequencing.&#xD;
&#xD;
      AIM To investigate whether gut microbiota is altered in patients with AN. If confirmed, this&#xD;
      potentially may play a role in the pathogenesis, promoting susceptibility to development&#xD;
      and/or maintenance of AN.&#xD;
&#xD;
      HYPOTHESIS Patients with AN harbor an altered gut microbiota composition and functional&#xD;
      potential as determined by quantitative metagenomic analyses of microbial DNA isolated from&#xD;
      stool samples and sequenced applying a combination of deep and untargeted shotgun sequencing.&#xD;
&#xD;
      STUDY DESIGN A cross sectional multicenter study where the gut microbiome profiles in women&#xD;
      with AN are compared with age matched healthy, normal weight women.&#xD;
&#xD;
      SUBJECTS The patients are characterized according to the national guidelines. The following&#xD;
      data will be extracted from the medical files and included in the descriptive and&#xD;
      multivariate analyzes.&#xD;
&#xD;
      Clinical variables: Age, disease duration, BMI, %weight change during the last 4 weeks,&#xD;
      in/out patient status, medication, somatic and psychiatric comorbidity, smoking status.&#xD;
&#xD;
      Psychometric variables: Eating Disorder Inventory (EDI)-3.&#xD;
&#xD;
      Control subjects: The control subjects will be matched as described, and allocated from&#xD;
      siblings of patients, local health workers, or if necessary, from announcement at an official&#xD;
      Danish website for participation in clinical trials. The age matched women fill in a 17 items&#xD;
      health related questionnaire. Any chronic disease, medication or a history of psychiatric&#xD;
      illness results in their exclusion from participation. They are, however, allowed to use oral&#xD;
      contraceptives.&#xD;
&#xD;
      FECAL SAMPLING Fecal samples are collected in order to characterize the composition of the&#xD;
      gut microbiome. A sample of about 5 g is collected with the help of a collection tray.&#xD;
      Participants are instructed in immediately storing the sample in their freezer at -18°C.&#xD;
      Transportation is done at cold temperatures in an isolated box containing frozen elements,&#xD;
      and handed over to study personnel maximal 24 hours after delivery where it is stored at -80&#xD;
      C until DNA extraction.&#xD;
&#xD;
      MORNING FASTING URINE 5 ml morning fasting urine is collected.&#xD;
&#xD;
      ANALYZING FOR DIFFERENCES IN GUT MICROBIOTA COMPOSITION The feces specimens are processed and&#xD;
      analyzed as follows: Bacterial DNA extraction, shotgun illumine DNA sequencing, microbial&#xD;
      gene analyses, taxonomy analyses including entero types, known species and unknown&#xD;
      meta-species (=quantitative metagenomics) as well as functional gene annotation. The DNA&#xD;
      purification and the shotgun sequencing is done at Unité MetaGenoPolis - Centre de recherche&#xD;
      de Jouy-en-Josas, MGP-INRA, JOUY en JOSAS, FRANCE. Quantitative metagenomics as outlined&#xD;
      above is performed by the unit of Bioinformatics and Computational Metagenomics at CBMR&#xD;
      applying established state-of-the art methodologies.&#xD;
&#xD;
      BIOCHEMICAL VARIABLES Basic parameters: The following analyses are performed as an integral&#xD;
      part of diagnosis and treatment in specialist units, and will be extracted from the medical&#xD;
      files: Na, K, creatinine, carbamide, bicarbonate, Ca-ion, phosphate, Hemoglobin, C-reactive&#xD;
      protein, leukocytes, mean cell volume, mean cell hemoglobin concentrate, alanine&#xD;
      aminotransferase, alkaline phosphatase, bilirubin, pancreatic amylase, Mg, Zn, thyroxine,&#xD;
      triiodothyronine, thyrotropin, cobalamine, folate, follicle stimulating hormone, luteinizing&#xD;
      hormone, prolactin, estradiol, and vitamin D. Values will be adjusted and expressed as ratio&#xD;
      relative to the normal value on the basis of average standard values for each centers.&#xD;
&#xD;
      SAMPLES&#xD;
&#xD;
      All samples are stored at -80 C:&#xD;
&#xD;
        -  Faeces 4 x 5 g.&#xD;
&#xD;
        -  Serum 20 x 500 microliter serum.&#xD;
&#xD;
        -  Plasma 20 x 500 microliter plasm.&#xD;
&#xD;
        -  Buffy coat x 10.&#xD;
&#xD;
        -  Morning urine 2 ml.&#xD;
&#xD;
      Biochemical analyses: From the sampled serum/plasma, following analyses are performed:&#xD;
      Insulin, Insulin-like growth factor-I (IGF-I), IGF-1 binding protein-3, leptin, and&#xD;
      adiponectin.&#xD;
&#xD;
      BIOBANK The fecal samples and the above mentioned materials will be stored in a biobank in up&#xD;
      to 30 years. If a microbiome genotype for AN is disclosed, further genomic analyses, mass&#xD;
      spectroscopy for metabolites and maybe inoculation studies will be performed (followed by new&#xD;
      applications to the Ethical Committee).&#xD;
&#xD;
      STATISTICS Power calculation. There is no data available regarding microbiota in AN. The&#xD;
      number of participants is based on ongoing studies of gut microbiome in pregnant women and&#xD;
      patients with multiple sclerosis where a population size of 50 was estimated to be&#xD;
      appropriate for providing a power greater than 80% and a significance of 5% for observing&#xD;
      differences in the present quantitative metagenomics approach.&#xD;
&#xD;
      Multivariate analyze. Statistical methods will build upon previously developed methods and&#xD;
      multivariate statistical approaches applied within the Metagenomics of the Human Intestinal&#xD;
      Tract (MetaHIT) consortium.&#xD;
&#xD;
      ETHICS The study is approved and registered at The Regional Scientific Ethical Committee for&#xD;
      Southern Denmark (file no 42053 S-20140040).&#xD;
&#xD;
      The participants will be asked whether they are willing to donate the material (feces, blood,&#xD;
      urine) to a biobank for later analysis, and will only be used in related future studies, for&#xD;
      which approval will first be obtained from the Scientific Ethical Committee. The biobank&#xD;
      samples are stored up to 30 years. Clinical, psychometric and biochemical variables mentioned&#xD;
      above are extracted from medical files. Patients from the three participating centres will&#xD;
      receive an invitation to an information meeting with one of the health workers, who are one&#xD;
      of the local investigators. For subjects below age of 18 years (15 - 18 y) one or both&#xD;
      parents must participate and must provide written informed consent. Subjects above age 18&#xD;
      years are allowed to bring a friend or family member as an observer. It will be emphasized&#xD;
      that it is voluntary to participate. During the information meeting an official document&#xD;
      published by The National Committee on Health Research Ethics 2012 is handed.&#xD;
&#xD;
      The invitation of siblings is motivated by the fact that future studies of biobank material&#xD;
      can prove to be of particular interest, if there is a high level of genetic relatedness&#xD;
      between the test patients and the control subjects. However, the patient inclusion in the&#xD;
      present project will not depend on whether there are siblings who wish to participate.&#xD;
&#xD;
      Recruitment of patients down to the age of 15 are motivated by the fact that the highest&#xD;
      prevalence of AN is in this age group. The older the patient, the greater is the likelihood&#xD;
      for psychiatric co-morbidity. It therefore will be important for the validity of the results&#xD;
      that the age spectrum also covers 15 - 18 years of age.&#xD;
&#xD;
      HANDLING AND ARCHIVING DATA The current protocol are approved by The Danish Data Protection&#xD;
      Agency (case files 1429534, 2008-58-035) and all the documents and materials related to the&#xD;
      clinical study will be stored according to the Act on Processing of Personal Data. All&#xD;
      sensitive information of personal, physical or biological origin related to the individual&#xD;
      participants will be treated confidentially according to the Danish legislation.&#xD;
&#xD;
      FINANCE This is a non-profit study, completely independent from commercial interests. There&#xD;
      will be applied for public and private grants to finance this study.&#xD;
&#xD;
      PERSPECTIVES If the present study identifies an altered gut flora in AN, it provides the&#xD;
      basis for several important new studies of gen products in faces and blood in AN, and also&#xD;
      inoculation experiments with gnotobiotic mice. Clarifying whether the intestinal flora is&#xD;
      implicated in the susceptibility to or maintenance of AN may provide the basis for&#xD;
      development of new highly required treatments of AN and potentially also obesity.&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Active, not recruiting</overall_status>
  <start_date type="Actual">September 1, 2014</start_date>
  <completion_date type="Anticipated">December 2022</completion_date>
  <primary_completion_date type="Actual">September 1, 2016</primary_completion_date>
  <study_type>Observational</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <observational_model>Case-Control</observational_model>
    <time_perspective>Cross-Sectional</time_perspective>
  </study_design_info>
  <primary_outcome>
    <measure>Diversity of faecal microbiota</measure>
    <time_frame>Cross-sectional study with only one time point. The sample is delivered at a convenient time during an ongoing psychotherapeutic treatment.</time_frame>
    <description>Metagenomic analyses of microbial DNA isolated from stool samples and sequenced applying a combination of deep and untargeted shotgun sequencing.</description>
  </primary_outcome>
  <enrollment type="Actual">151</enrollment>
  <condition>Anorexia Nervosa</condition>
  <biospec_retention>Samples With DNA</biospec_retention>
  <biospec_descr>
    <textblock>
      Gut microbiome Serum and plasma Cells (buffy coat) Urine&#xD;
    </textblock>
  </biospec_descr>
  <eligibility>
    <study_pop>
      <textblock>
        Patients referred to specialist unit.&#xD;
      </textblock>
    </study_pop>
    <sampling_method>Probability Sample</sampling_method>
    <criteria>
      <textblock>
        Inclusion:&#xD;
&#xD;
          -  Women.&#xD;
&#xD;
          -  Referred to specialized eating disorder center.&#xD;
&#xD;
          -  Fulfill criteria in Diagnostic and Statistical Manual of Mental Disorders - 5.&#xD;
&#xD;
        Exclusion:&#xD;
&#xD;
          -  Age &lt;15 years.&#xD;
&#xD;
          -  Known ongoing abuse of laxatives.&#xD;
&#xD;
          -  Antibiotic treatment within the last three months.&#xD;
&#xD;
          -  Inflammatory bowel disease, colorectal cancer or other bowel disease.&#xD;
      </textblock>
    </criteria>
    <gender>Female</gender>
    <minimum_age>15 Years</minimum_age>
    <maximum_age>N/A</maximum_age>
    <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>René K Støving, PhD</last_name>
    <role>Principal Investigator</role>
    <affiliation>Center for Eating Disorders, Odense University Hospital</affiliation>
  </overall_official>
  <location>
    <facility>
      <name>Unit for Psychiatric Research, Aalborg University Hospital.</name>
      <address>
        <city>Aalborg</city>
        <zip>9000</zip>
        <country>Denmark</country>
      </address>
    </facility>
  </location>
  <location>
    <facility>
      <name>Child and adolescent psychiatric unit. Risskov. Aarhus University Hospital.</name>
      <address>
        <city>Aarhus</city>
        <zip>8000</zip>
        <country>Denmark</country>
      </address>
    </facility>
  </location>
  <location>
    <facility>
      <name>Department of Endocrinology, Center for Eating Disorders. Odense University Hospital.</name>
      <address>
        <city>Odense</city>
        <zip>5000</zip>
        <country>Denmark</country>
      </address>
    </facility>
  </location>
  <location_countries>
    <country>Denmark</country>
  </location_countries>
  <verification_date>April 2022</verification_date>
  <study_first_submitted>July 29, 2014</study_first_submitted>
  <study_first_submitted_qc>August 13, 2014</study_first_submitted_qc>
  <study_first_posted type="Estimate">August 15, 2014</study_first_posted>
  <last_update_submitted>September 24, 2022</last_update_submitted>
  <last_update_submitted_qc>September 24, 2022</last_update_submitted_qc>
  <last_update_posted type="Actual">September 27, 2022</last_update_posted>
  <responsible_party>
    <responsible_party_type>Principal Investigator</responsible_party_type>
    <investigator_affiliation>Odense University Hospital</investigator_affiliation>
    <investigator_full_name>René Klinkby Støving</investigator_full_name>
    <investigator_title>research secretary</investigator_title>
  </responsible_party>
  <keyword>Microbiome</keyword>
  <keyword>DNA sequencing</keyword>
  <keyword>Leptin</keyword>
  <keyword>Adiponectin</keyword>
  <keyword>Insulin</keyword>
  <keyword>IGF-I</keyword>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Anorexia</mesh_term>
    <mesh_term>Anorexia Nervosa</mesh_term>
  </condition_browse>
  <patient_data>
    <sharing_ipd>Undecided</sharing_ipd>
  </patient_data>
  <!-- Results have not yet been posted for this study                                          -->
</clinical_study>


download date: ClinicalTrials.gov processed this data on September 20, 2023
link text: Link to the current ClinicalTrials.gov record.
url: https://clinicaltrials.gov/ct2/show/NCT02217384
org study id: S-20140040
nct id: NCT02217384
lead sponsor: 
collaborator: 
collaborator: 
collaborator: 
collaborator: 
has dmc: Yes
Background. Anorexia nervosa (AN) still carries the highest fatality rate of any psychiatric
      disease, and less than half of the patients recover, completely refractory to any treatment.
      The etiology remains unknown and evidence for treatment is lacking. The intestinal microbiota
      and its microbiome provide humans with additional gene products which may be regarded as an
      organ, which contributes to multiple host metabolic pathways. Recent advances in microbial
      DNA sequencing technologies have resulted in metagenomic DNA analysis of whole ecosystems
      such as the human gut. Altered intestinal microbiota has been related to obesity and insulin
      resistance. Hypothetically, the intestinal microbiota could play a role in the generation
      and/or maintenance of the emaciation in AN.

      Aim. The aim of the present study is to investigate whether gut microbiota is altered in
      patients with AN.

      Subjects and methods. A cross sectional study of the gut microbiome profiles in 75 clinical,
      psychometric and biochemical well characterized treatment seeking females with AN. The
      microbiome profiles are compared with 75 age- matched healthy Danish control subjects.

      Perspectives. Clarifying whether the intestinal flora is implicated in the susceptibility to
      or maintenance of AN may provide the basis for development of new highly required treatments.
BACKGROUND Anorexia nervosa (AN) is a complex disorder characterized by disturbed body image,
      ego-syntonic neglect, ambivalence, self-starvation, loss of body weight, obsessive thoughts
      of food, ritualistic patterns of food intake, elevated physical activity, depression, anxiety
      and emotional rigidity. The evidence base for the treatment is very limited, considering the
      extent to which this disorder erodes quality of life and still carries the highest fatality
      rate of any psychiatric diseases. In fact, there is no evidence that the prognosis has
      improved throughout the 20th century.

      AN remains a syndrome, i.e. collections of symptoms, as it is not defined by etiology. Twin
      studies have consistently demonstrated that AN is strongly familial predominantly due to
      genetic factors. However, neither candidate gene nor genome-wide studies have identified
      truly validated genes for AN. AN is associated with multiple, profound endocrine
      disturbances. However, so far the extensive research has not led to any breakthrough
      discovery. There is a need for new thinking.

      Colonization of the human gut begins at birth with bacteria from the mother's vagina and
      labor-released maternal feces. In adults the human digestive tract contains complex
      assemblies of microorganisms which actually outnumber the number of the host cells 10:1. This
      intestinal microbiota and its microbiome provide humans with additional gene products, which
      may be regarded as an organ, which contributes to multiple host metabolic pathways. Thus, a
      whole range of substances are produced which are not yet identified. Potentially some of
      these substances may affect functions in the brain, such as appetite and emotion regulations.
      Recent advances in microbial DNA sequencing technologies have resulted in metagenomics DNA
      analysis of whole ecosystems such as the human gut. An important step forward in
      investigating the bacterial composition of the human microbiome was taken with the Metagenome
      of the Human Intestinal Tract consortium reporting the first complete microbial gene atlas of
      the human distal gut, using deep next generation metagenome sequencing.

      AIM To investigate whether gut microbiota is altered in patients with AN. If confirmed, this
      potentially may play a role in the pathogenesis, promoting susceptibility to development
      and/or maintenance of AN.

      HYPOTHESIS Patients with AN harbor an altered gut microbiota composition and functional
      potential as determined by quantitative metagenomic analyses of microbial DNA isolated from
      stool samples and sequenced applying a combination of deep and untargeted shotgun sequencing.

      STUDY DESIGN A cross sectional multicenter study where the gut microbiome profiles in women
      with AN are compared with age matched healthy, normal weight women.

      SUBJECTS The patients are characterized according to the national guidelines. The following
      data will be extracted from the medical files and included in the descriptive and
      multivariate analyzes.

      Clinical variables: Age, disease duration, BMI, %weight change during the last 4 weeks,
      in/out patient status, medication, somatic and psychiatric comorbidity, smoking status.

      Psychometric variables: Eating Disorder Inventory (EDI)-3.

      Control subjects: The control subjects will be matched as described, and allocated from
      siblings of patients, local health workers, or if necessary, from announcement at an official
      Danish website for participation in clinical trials. The age matched women fill in a 17 items
      health related questionnaire. Any chronic disease, medication or a history of psychiatric
      illness results in their exclusion from participation. They are, however, allowed to use oral
      contraceptives.

      FECAL SAMPLING Fecal samples are collected in order to characterize the composition of the
      gut microbiome. A sample of about 5 g is collected with the help of a collection tray.
      Participants are instructed in immediately storing the sample in their freezer at -18°C.
      Transportation is done at cold temperatures in an isolated box containing frozen elements,
      and handed over to study personnel maximal 24 hours after delivery where it is stored at -80
      C until DNA extraction.

      MORNING FASTING URINE 5 ml morning fasting urine is collected.

      ANALYZING FOR DIFFERENCES IN GUT MICROBIOTA COMPOSITION The feces specimens are processed and
      analyzed as follows: Bacterial DNA extraction, shotgun illumine DNA sequencing, microbial
      gene analyses, taxonomy analyses including entero types, known species and unknown
      meta-species (=quantitative metagenomics) as well as functional gene annotation. The DNA
      purification and the shotgun sequencing is done at Unité MetaGenoPolis - Centre de recherche
      de Jouy-en-Josas, MGP-INRA, JOUY en JOSAS, FRANCE. Quantitative metagenomics as outlined
      above is performed by the unit of Bioinformatics and Computational Metagenomics at CBMR
      applying established state-of-the art methodologies.

      BIOCHEMICAL VARIABLES Basic parameters: The following analyses are performed as an integral
      part of diagnosis and treatment in specialist units, and will be extracted from the medical
      files: Na, K, creatinine, carbamide, bicarbonate, Ca-ion, phosphate, Hemoglobin, C-reactive
      protein, leukocytes, mean cell volume, mean cell hemoglobin concentrate, alanine
      aminotransferase, alkaline phosphatase, bilirubin, pancreatic amylase, Mg, Zn, thyroxine,
      triiodothyronine, thyrotropin, cobalamine, folate, follicle stimulating hormone, luteinizing
      hormone, prolactin, estradiol, and vitamin D. Values will be adjusted and expressed as ratio
      relative to the normal value on the basis of average standard values for each centers.

      SAMPLES

      All samples are stored at -80 C:

        -  Faeces 4 x 5 g.

        -  Serum 20 x 500 microliter serum.

        -  Plasma 20 x 500 microliter plasm.

        -  Buffy coat x 10.

        -  Morning urine 2 ml.

      Biochemical analyses: From the sampled serum/plasma, following analyses are performed:
      Insulin, Insulin-like growth factor-I (IGF-I), IGF-1 binding protein-3, leptin, and
      adiponectin.

      BIOBANK The fecal samples and the above mentioned materials will be stored in a biobank in up
      to 30 years. If a microbiome genotype for AN is disclosed, further genomic analyses, mass
      spectroscopy for metabolites and maybe inoculation studies will be performed (followed by new
      applications to the Ethical Committee).

      STATISTICS Power calculation. There is no data available regarding microbiota in AN. The
      number of participants is based on ongoing studies of gut microbiome in pregnant women and
      patients with multiple sclerosis where a population size of 50 was estimated to be
      appropriate for providing a power greater than 80% and a significance of 5% for observing
      differences in the present quantitative metagenomics approach.

      Multivariate analyze. Statistical methods will build upon previously developed methods and
      multivariate statistical approaches applied within the Metagenomics of the Human Intestinal
      Tract (MetaHIT) consortium.

      ETHICS The study is approved and registered at The Regional Scientific Ethical Committee for
      Southern Denmark (file no 42053 S-20140040).

      The participants will be asked whether they are willing to donate the material (feces, blood,
      urine) to a biobank for later analysis, and will only be used in related future studies, for
      which approval will first be obtained from the Scientific Ethical Committee. The biobank
      samples are stored up to 30 years. Clinical, psychometric and biochemical variables mentioned
      above are extracted from medical files. Patients from the three participating centres will
      receive an invitation to an information meeting with one of the health workers, who are one
      of the local investigators. For subjects below age of 18 years (15 - 18 y) one or both
      parents must participate and must provide written informed consent. Subjects above age 18
      years are allowed to bring a friend or family member as an observer. It will be emphasized
      that it is voluntary to participate. During the information meeting an official document
      published by The National Committee on Health Research Ethics 2012 is handed.

      The invitation of siblings is motivated by the fact that future studies of biobank material
      can prove to be of particular interest, if there is a high level of genetic relatedness
      between the test patients and the control subjects. However, the patient inclusion in the
      present project will not depend on whether there are siblings who wish to participate.

      Recruitment of patients down to the age of 15 are motivated by the fact that the highest
      prevalence of AN is in this age group. The older the patient, the greater is the likelihood
      for psychiatric co-morbidity. It therefore will be important for the validity of the results
      that the age spectrum also covers 15 - 18 years of age.

      HANDLING AND ARCHIVING DATA The current protocol are approved by The Danish Data Protection
      Agency (case files 1429534, 2008-58-035) and all the documents and materials related to the
      clinical study will be stored according to the Act on Processing of Personal Data. All
      sensitive information of personal, physical or biological origin related to the individual
      participants will be treated confidentially according to the Danish legislation.

      FINANCE This is a non-profit study, completely independent from commercial interests. There
      will be applied for public and private grants to finance this study.

      PERSPECTIVES If the present study identifies an altered gut flora in AN, it provides the
      basis for several important new studies of gen products in faces and blood in AN, and also
      inoculation experiments with gnotobiotic mice. Clarifying whether the intestinal flora is
      implicated in the susceptibility to or maintenance of AN may provide the basis for
      development of new highly required treatments of AN and potentially also obesity.
observational model: Case-Control
time perspective: Cross-Sectional
measure: Diversity of faecal microbiota
time frame: Cross-sectional study with only one time point. The sample is delivered at a convenient time during an ongoing psychotherapeutic treatment.
description: Metagenomic analyses of microbial DNA isolated from stool samples and sequenced applying a combination of deep and untargeted shotgun sequencing.
Gut microbiome Serum and plasma Cells (buffy coat) Urine
study pop: 
sampling method: Probability Sample
criteria: 
gender: Female
minimum age: 15 Years
maximum age: N/A
healthy volunteers: Accepts Healthy Volunteers
last name: René K Støving, PhD
role: Principal Investigator
affiliation: Center for Eating Disorders, Odense University Hospital
facility: 
facility: 
facility: 
country: Denmark
responsible party type: Principal Investigator
investigator affiliation: Odense University Hospital
investigator full name: René Klinkby Støving
investigator title: research secretary
mesh term: Anorexia
mesh term: Anorexia Nervosa
sharing ipd: Undecided