<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/NCT02969759</url>
  </required_header>
  <id_info>
    <org_study_id>2016/36</org_study_id>
    <nct_id>NCT02969759</nct_id>
  </id_info>
  <brief_title>Bioenergetics and Protein Metabolism in Sporadic Amyotrophic Lateral Sclerosis</brief_title>
  <acronym>Fibro-ALS</acronym>
  <official_title>Bioenergetics and Protein Metabolism in Sporadic Amyotrophic Lateral Sclerosis</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>University Hospital, Angers</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
  </sponsors>
  <source>University Hospital, Angers</source>
  <brief_summary>
    <textblock>
      INTRODUCTION: Amyotrophic Lateral Sclerosis (ALS) is a degenerative disease of the motor&#xD;
      neurones of the brain and the spinal cord. The pathophysiological mechanisms of the disease&#xD;
      remain unknown. The average age of onset of ALS is about 60 years old, and the mean survival&#xD;
      of patients is about 2 years. The disease is responsible for a progressive paralysis leading&#xD;
      to death from respiratory failure. The only treatment available is the Riluzole, with a very&#xD;
      modest efficiency on the progression of the disease. ALS is the third neurodegenerative&#xD;
      disease, affecting 6000 persons in France, 150 000 in the world. Among the protagonists&#xD;
      involved in the occurrence of the disease, mitochondrial perturbations and protein&#xD;
      accumulations seem to be central elements.&#xD;
&#xD;
      OBJECTIVES: To precise the implication of energy and protein metabolism in the sporadic forms&#xD;
      of ALS, to identify potential biomarkers of the disease and to test new therapies.&#xD;
&#xD;
      METHODS: The investigators will study cell growth, bioenergetics, mitochondrial dynamics,&#xD;
      free-radicals production, presence of cytoplasmic inclusions, cytoskeleton structure and&#xD;
      stress response in primary skin fibroblasts obtained from sporadic ALS patients. The study&#xD;
      will be conducted over a period of three years in 3 centres specialized respectively in motor&#xD;
      neuron diseases, mitochondrial metabolism and neuronal cytoskeleton.&#xD;
&#xD;
      PROSPECTS: If the investigators achieve to identify differences between ALS fibroblasts and&#xD;
      controls, the results will be key elements to reinforce the hypothesis of a systemic disease&#xD;
      with an important metabolic participation, to better define ALS pathophysiological&#xD;
      mechanisms, to find potential biomarkers and to test new therapies.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      The investigators will study fibroblasts derived from sporadic ALS patients.&#xD;
&#xD;
      Clinical parameters will be collected such as sex, age, age of onset, clinical presentation,&#xD;
      electrophysiological an respiratory parameters, treatments.&#xD;
&#xD;
      Patients and controls derived fibroblasts will be amplified and cultivated from skin&#xD;
      biopsies.&#xD;
&#xD;
      Primary fibroblast cell cultures will be derived from skin biopsies obtained from control&#xD;
      subjects and from the patients after obtaining written consent. Amplifications will be&#xD;
      performed, from passage 1 to 6, allowing the time for cell cryopreservation of 10 samples per&#xD;
      cell line. All experiments will be conducted on cells with similar passage numbers, ranging&#xD;
      from 6 to 25, to avoid artefacts due to senescence.&#xD;
&#xD;
      Cell growth of each line will be monitored with the IncuCyte ZOOM® system (Essen&#xD;
      Biosciences), a microscope gantry that resides in a cell incubator, and a networked external&#xD;
      controller hard drive that gathers and processes image data. Cell growth will be&#xD;
      automatically calculated at each time point using the relative cell density (ratio of the&#xD;
      occupied area to the total area of the acquired regions).&#xD;
&#xD;
      The respiratory parameters of the fibroblasts will be monitored precisely with an high&#xD;
      resolution oxygraph (OROBOROS). Enzymatic activity of each complex of the mitochondrial&#xD;
      respiratory chain will be measured by spectrometry, and the mitochondrial membrane potential&#xD;
      will be studied with fluorescence microscopy (TMRM probe). The investigators will also study&#xD;
      mitochondrial dynamics and the architecture of mitochondrial network with fluorescence&#xD;
      microscopy and super-resolution microscopy (SRM, Roper). The production of free radicals will&#xD;
      be estimated with probes sensitive to oxidative stress. Finally, a metabolomic study (mass&#xD;
      spectrometry QTRAP LC/MS/MS 5500 ABSciex) will be performed to characterise precisely the&#xD;
      metabolites in the cells and to assess their metabolic profile.&#xD;
&#xD;
      The investigators will search for protein inclusions (intermediate filaments, TDP43, FUS,&#xD;
      ubiquitin) with immuno-fluorescence technics. Western Blots will be performed to study the&#xD;
      protein metabolism. The architecture of the cytoskeleton will be analysed in SRM and&#xD;
      fluorescence microscopy, and the investigators will try to transfect fibroblasts in order to&#xD;
      express Neurofilaments and study the consequences on the cytoskeleton and protein metabolism.&#xD;
&#xD;
      The investigators will study the response of the cells to the stress (hypoxia SCI-tive Dual,&#xD;
      starvation), and to senescence (high passage numbers).&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Unknown status</overall_status>
  <last_known_status>Not yet recruiting</last_known_status>
  <start_date>November 2016</start_date>
  <completion_date type="Anticipated">November 2019</completion_date>
  <primary_completion_date type="Anticipated">November 2017</primary_completion_date>
  <phase>Early Phase 1</phase>
  <study_type>Interventional</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <allocation>Non-Randomized</allocation>
    <intervention_model>Parallel Assignment</intervention_model>
    <primary_purpose>Basic Science</primary_purpose>
    <masking>None (Open Label)</masking>
  </study_design_info>
  <primary_outcome>
    <measure>Kinetics of fibroblast growth</measure>
    <time_frame>24 hours</time_frame>
    <description>The growth rates of the fibroblast lines</description>
  </primary_outcome>
  <secondary_outcome>
    <measure>Mitochondrial metabolism</measure>
    <time_frame>24 hours</time_frame>
    <description>the breath parameters of fibroblast will be precisely studied by oxygraphy, a method consisting in measuring the oxygen consumption rates of fibroblasts in different metabolic phases</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Protéic metabolism</measure>
    <time_frame>24 hours</time_frame>
    <description>The presence of intracytoplasmic inclusion of target protein will be sought.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>stress in senescence answer</measure>
    <time_frame>24 hours</time_frame>
    <description>the answer due to the stress of cell lines in hypoxia and nutritionnal lack will be studied.</description>
  </secondary_outcome>
  <number_of_arms>2</number_of_arms>
  <enrollment type="Anticipated">30</enrollment>
  <condition>Sporadic Amyotrophic Lateral Sclerosis</condition>
  <arm_group>
    <arm_group_label>Patients</arm_group_label>
    <arm_group_type>Other</arm_group_type>
    <description>ALS defined by El Escorial Criteria.</description>
  </arm_group>
  <arm_group>
    <arm_group_label>Controls</arm_group_label>
    <arm_group_type>Other</arm_group_type>
    <description>Asymptomatic subjects with normal examination.</description>
  </arm_group>
  <intervention>
    <intervention_type>Other</intervention_type>
    <intervention_name>Skin Biopsy</intervention_name>
    <arm_group_label>Controls</arm_group_label>
    <arm_group_label>Patients</arm_group_label>
  </intervention>
  <eligibility>
    <criteria>
      <textblock>
        Inclusion criteria :&#xD;
&#xD;
          -  a free signed an written consent&#xD;
&#xD;
          -  aged between 18 and 80 years old&#xD;
&#xD;
          -  ALS defined by El Escorial Criteria&#xD;
&#xD;
          -  normal neurological examination&#xD;
&#xD;
        Exclusion criteria:&#xD;
&#xD;
          -  comorbidity or treatment susceptible to impact on metabolism&#xD;
&#xD;
          -  differential diagnosis suspected, early and late forms (&lt;6 month &gt; 3 years)&#xD;
&#xD;
          -  ALS not defined by El Escorial Criteria&#xD;
&#xD;
          -  withdrawal of consent to participate&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>18 Years</minimum_age>
    <maximum_age>80 Years</maximum_age>
    <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
  </eligibility>
  <verification_date>November 2016</verification_date>
  <study_first_submitted>November 4, 2016</study_first_submitted>
  <study_first_submitted_qc>November 17, 2016</study_first_submitted_qc>
  <study_first_posted type="Estimate">November 21, 2016</study_first_posted>
  <last_update_submitted>November 22, 2016</last_update_submitted>
  <last_update_submitted_qc>November 22, 2016</last_update_submitted_qc>
  <last_update_posted type="Estimate">November 23, 2016</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor</responsible_party_type>
  </responsible_party>
  <keyword>Bioenergetics</keyword>
  <keyword>Protein Metabolism</keyword>
  <keyword>Sporadic Amyotrophic Lateral Sclerosis</keyword>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Motor Neuron Disease</mesh_term>
    <mesh_term>Amyotrophic Lateral Sclerosis</mesh_term>
    <mesh_term>Sclerosis</mesh_term>
  </condition_browse>
  <!-- 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/NCT02969759
org study id: 2016/36
nct id: NCT02969759
lead sponsor: 
INTRODUCTION: Amyotrophic Lateral Sclerosis (ALS) is a degenerative disease of the motor
      neurones of the brain and the spinal cord. The pathophysiological mechanisms of the disease
      remain unknown. The average age of onset of ALS is about 60 years old, and the mean survival
      of patients is about 2 years. The disease is responsible for a progressive paralysis leading
      to death from respiratory failure. The only treatment available is the Riluzole, with a very
      modest efficiency on the progression of the disease. ALS is the third neurodegenerative
      disease, affecting 6000 persons in France, 150 000 in the world. Among the protagonists
      involved in the occurrence of the disease, mitochondrial perturbations and protein
      accumulations seem to be central elements.

      OBJECTIVES: To precise the implication of energy and protein metabolism in the sporadic forms
      of ALS, to identify potential biomarkers of the disease and to test new therapies.

      METHODS: The investigators will study cell growth, bioenergetics, mitochondrial dynamics,
      free-radicals production, presence of cytoplasmic inclusions, cytoskeleton structure and
      stress response in primary skin fibroblasts obtained from sporadic ALS patients. The study
      will be conducted over a period of three years in 3 centres specialized respectively in motor
      neuron diseases, mitochondrial metabolism and neuronal cytoskeleton.

      PROSPECTS: If the investigators achieve to identify differences between ALS fibroblasts and
      controls, the results will be key elements to reinforce the hypothesis of a systemic disease
      with an important metabolic participation, to better define ALS pathophysiological
      mechanisms, to find potential biomarkers and to test new therapies.
The investigators will study fibroblasts derived from sporadic ALS patients.

      Clinical parameters will be collected such as sex, age, age of onset, clinical presentation,
      electrophysiological an respiratory parameters, treatments.

      Patients and controls derived fibroblasts will be amplified and cultivated from skin
      biopsies.

      Primary fibroblast cell cultures will be derived from skin biopsies obtained from control
      subjects and from the patients after obtaining written consent. Amplifications will be
      performed, from passage 1 to 6, allowing the time for cell cryopreservation of 10 samples per
      cell line. All experiments will be conducted on cells with similar passage numbers, ranging
      from 6 to 25, to avoid artefacts due to senescence.

      Cell growth of each line will be monitored with the IncuCyte ZOOM® system (Essen
      Biosciences), a microscope gantry that resides in a cell incubator, and a networked external
      controller hard drive that gathers and processes image data. Cell growth will be
      automatically calculated at each time point using the relative cell density (ratio of the
      occupied area to the total area of the acquired regions).

      The respiratory parameters of the fibroblasts will be monitored precisely with an high
      resolution oxygraph (OROBOROS). Enzymatic activity of each complex of the mitochondrial
      respiratory chain will be measured by spectrometry, and the mitochondrial membrane potential
      will be studied with fluorescence microscopy (TMRM probe). The investigators will also study
      mitochondrial dynamics and the architecture of mitochondrial network with fluorescence
      microscopy and super-resolution microscopy (SRM, Roper). The production of free radicals will
      be estimated with probes sensitive to oxidative stress. Finally, a metabolomic study (mass
      spectrometry QTRAP LC/MS/MS 5500 ABSciex) will be performed to characterise precisely the
      metabolites in the cells and to assess their metabolic profile.

      The investigators will search for protein inclusions (intermediate filaments, TDP43, FUS,
      ubiquitin) with immuno-fluorescence technics. Western Blots will be performed to study the
      protein metabolism. The architecture of the cytoskeleton will be analysed in SRM and
      fluorescence microscopy, and the investigators will try to transfect fibroblasts in order to
      express Neurofilaments and study the consequences on the cytoskeleton and protein metabolism.

      The investigators will study the response of the cells to the stress (hypoxia SCI-tive Dual,
      starvation), and to senescence (high passage numbers).
allocation: Non-Randomized
intervention model: Parallel Assignment
primary purpose: Basic Science
masking: None (Open Label)
measure: Kinetics of fibroblast growth
time frame: 24 hours
description: The growth rates of the fibroblast lines
measure: Mitochondrial metabolism
time frame: 24 hours
description: the breath parameters of fibroblast will be precisely studied by oxygraphy, a method consisting in measuring the oxygen consumption rates of fibroblasts in different metabolic phases
measure: Protéic metabolism
time frame: 24 hours
description: The presence of intracytoplasmic inclusion of target protein will be sought.
measure: stress in senescence answer
time frame: 24 hours
description: the answer due to the stress of cell lines in hypoxia and nutritionnal lack will be studied.
arm group label: Patients
arm group type: Other
description: ALS defined by El Escorial Criteria.
arm group label: Controls
arm group type: Other
description: Asymptomatic subjects with normal examination.
intervention type: Other
intervention name: Skin Biopsy
arm group label: Controls
arm group label: Patients
criteria: 
gender: All
minimum age: 18 Years
maximum age: 80 Years
healthy volunteers: Accepts Healthy Volunteers
responsible party type: Sponsor
mesh term: Motor Neuron Disease
mesh term: Amyotrophic Lateral Sclerosis
mesh term: Sclerosis