<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/NCT02256241</url>
  </required_header>
  <id_info>
    <org_study_id>0180-14-RMB</org_study_id>
    <nct_id>NCT02256241</nct_id>
  </id_info>
  <brief_title>The Role of RING Ubiquitin Ligases in Biologic and Oncologic Processes in Tissues of Mesenchymal Origin</brief_title>
  <acronym>RING UB LIGASE</acronym>
  <official_title>The Role of RING Ubiquitin Ligases in Biologic and Oncologic Processes in Tissues of Mesenchymal Origin</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>Rambam Health Care Campus</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>Technion, Israel Institute of Technology</agency>
      <agency_class>Other</agency_class>
    </collaborator>
  </sponsors>
  <source>Rambam Health Care Campus</source>
  <oversight_info>
    <has_dmc>No</has_dmc>
  </oversight_info>
  <brief_summary>
    <textblock>
      Stage 1:&#xD;
&#xD;
      Bone marrow will be collected from otherwise healthy patients undergoing orthopedic surgery -&#xD;
      arthroplasty for the treatment of degenerative joint disease or traumatised patients. The&#xD;
      bone marrow will be collected from disposable tissue that is removed during the normal&#xD;
      sequence of the surgery. These cells will be used for:&#xD;
&#xD;
      Establishment of the role of RING ubiquitin ligases in osteogenic progenitors proliferation&#xD;
      and differentiation in culture.&#xD;
&#xD;
      I. To determine the levels of RING ubiquitin ligases mRNA and protein in differentiating&#xD;
      human mesenchymal precursor cells: To test the correlation between levels of RING ubiquitin&#xD;
      ligases and degree of osteogenic differentiation, we will extract mesenchymal precursor cells&#xD;
      from the collected tissue and test the mRNA protein level of RING ubiquitin ligases upon&#xD;
      induction of differentiation. Specifically, we will compare the levels of RING ubiquitin&#xD;
      ligases before and after the initiation of differentiational stimulus in time dependent&#xD;
      manner by western blot and real time PCR analysis.&#xD;
&#xD;
      II. To test the impact of specific RING ubiquitin ligases on differentiation of human&#xD;
      mesenchymal precursor cells: to test the impact of RING ubiquitin ligases on the&#xD;
      differentiation of mesenchymal precursor cell from the collected tissue by testing the&#xD;
      expression of classical differentiational bone markers by flow cytometry (fibronectin, CD105)&#xD;
      and by Alkaline phosphatase (ALP) activity assay. For this aim we developed both a&#xD;
      constitutive and Dox-regulated conditional overexpression and shRNA lenti-viral systems that&#xD;
      enables efficient modulation of these RING ubiquitin ligases level.&#xD;
&#xD;
      III. Determine the role of RING ubiquitin ligases in proliferation and survival of human&#xD;
      mesenchymal precursor cells: Via inhibition or overexpression of ligases in mesenchymal&#xD;
      progenitor cells we will test the role of these ligases in proliferation and survival of&#xD;
      mesenchymal precursors by using MTT assay, Propidion-Iodid (PI) and tunnel assays in flow&#xD;
      cytometry analysis.&#xD;
&#xD;
      Stage 2:&#xD;
&#xD;
      Collection of connective tissue from patients with malignancies of musculoskeletal origin.&#xD;
      The tissue that will be used is part of the resected tumor specimens. The tissue will be used&#xD;
      for:&#xD;
&#xD;
      The establishment of the role/s of RING ligases in musculoskeletal cancers using cell culture&#xD;
      and in vivo activation. Test if the expression of selected positive candidates from stage 1&#xD;
      correlates with cancer development and progression in human-derived samples Independently of&#xD;
      our mechanistic experiment we aim to determine the relevance of these ligases to human&#xD;
      musculoskeletal cancers. As the first step we will screen primary tumor biopsies at the&#xD;
      protein level correlates with cancer grade and prognosis. Toward this aim we recently&#xD;
      generated a highly specific several anti-monoclonal antibodies in our laboratory as well use&#xD;
      comercial available antibodies .&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      Stage 1:&#xD;
&#xD;
      Bone marrow will be collected from otherwise healthy patients undergoing orthopedic surgery -&#xD;
      arthroplasty for the treatment of degenerative joint disease or traumatised patients. The&#xD;
      bone marrow will be collected from disposable tissue that is removed during the normal&#xD;
      sequence of the surgery. These cells will be used for:&#xD;
&#xD;
      Establishment of the role of RING ubiquitin ligases in osteogenic progenitors proliferation&#xD;
      and differentiation in culture.&#xD;
&#xD;
      I. To determine the levels of RING ubiquitin ligases mRNA and protein in differentiating&#xD;
      human mesenchymal precursor cells: To test the correlation between levels of RING ubiquitin&#xD;
      ligases and degree of osteogenic differentiation, we will extract mesenchymal precursor cells&#xD;
      from the collected tissue and test the mRNA protein level of RING ubiquitin ligases upon&#xD;
      induction of differentiation. Specifically, we will compare the levels of RING ubiquitin&#xD;
      ligases RING ubiquitin ligases before and after the initiation of differentiational stimulus&#xD;
      in time dependent manner by western blot and real time PCR analysis.&#xD;
&#xD;
      II. To test the impact of RING ubiquitin ligases on differentiation of human mesenchymal&#xD;
      precursor cells: to test the impact of RING ubiquitin ligases on the differentiation of&#xD;
      mesenchymal precursor cell from the collected tissue by testing the expression of classical&#xD;
      differentiational bone markers by flow cytometry (fibronectin, CD105) and by Alkaline&#xD;
      phosphatase (ALP) activity assay. For this aim we developed both a constitutive and&#xD;
      Dox-regulated conditional RING ubiquitin ligases overexpression and shRNA lenti-viral systems&#xD;
      that enables efficient modulation of RING ubiquitin ligases level.&#xD;
&#xD;
      III. Determine the role of RING ubiquitin ligases in proliferation and survival of human&#xD;
      mesenchymal precursor cells: We will test the role of their in proliferation and survival of&#xD;
      mesenchymal precursors by using MTT assay, Propidion-Iodid (PI) and tunnel assays in flow&#xD;
      cytometry analysis.&#xD;
&#xD;
      Stage 2:&#xD;
&#xD;
      Collection of connective tissue from patients with malignancies of musculoskeletal origin.&#xD;
      The tissue that will be used is part of the resected tumor specimens. The tissue will be used&#xD;
      for:&#xD;
&#xD;
      The establishment of the role/s of RING ubiquitin ligases in musculoskeletal cancers using&#xD;
      cell culture and in vivo.&#xD;
&#xD;
      I. Test the role of our in proliferation and survival in musculoskeletal cancers: Using our&#xD;
      lentiviral systems we will test the biological impact of inhibition and overexpression in&#xD;
      musculoskeletal neoplastic cells.&#xD;
&#xD;
      II. Test whether inhibition or overexpression of these ligases impacts the biology of&#xD;
      musculoskeletal cancers.We will check if inhibition of these ligases in malignant cells of&#xD;
      musculoskeletal origin by lenti-viral infections induces differentiation. Infected cells will&#xD;
      be analyzed for expression of differentiational markers in flow cytometry and&#xD;
      immunohistochemistry.&#xD;
&#xD;
      III. Test if expression of RING ubiquitin ligases correlates with cancer development and&#xD;
      progression in human-derived samples Independently of our mechanistic experiment we aim to&#xD;
      determine the relevance of RING ubiquitin ligases to human musculoskeletal cancers.&#xD;
&#xD;
      Patients consent will be signed after properly informed at time of pre-surgical preparation&#xD;
      visit. In cases when pre-surgical visit is not held consent will be signed prior to the day&#xD;
      of surgery.&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Unknown status</overall_status>
  <last_known_status>Recruiting</last_known_status>
  <start_date>October 2014</start_date>
  <completion_date type="Anticipated">July 2019</completion_date>
  <primary_completion_date type="Anticipated">July 2019</primary_completion_date>
  <study_type>Observational</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <observational_model>Cohort</observational_model>
  </study_design_info>
  <primary_outcome>
    <measure>Protein and mRNA levels of RING ubiquitin ligases during osteogenesis</measure>
    <time_frame>5 years</time_frame>
    <description>To determine the levels of RING ubiquitin ligases mRNA and protein in differentiating human mesenchymal precursor cells. We will extract mesenchymal precursor cells from the collected tissue and test the mRNA and protein levels of RING ubiquitin ligases upon induction of differentiation.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Protein and mRNA levels of RING ubiquitin ligases in musculoskeletal tumors and its correlation to staging and grading</measure>
    <time_frame>Time Frame: * 5 years</time_frame>
    <description>It is not clear if RING ubiquitin ligases protein levels are elevated in soft tissue-derived cancers (primary and metastatic tumors) and whether there is a correlation between RING ubiquitin ligases levels and staging and grading of soft tissue-derived cancers. For this reason, we will check whether RING ubiquitin ligases expression correlates with cancer development and progression in human-derived samples. We will determine the relevance of RING ubiquitin ligases to human musculoskeletal cancers. To do this we will screen primary tumor biopsies for RNA and protein levels of RING ubiquitin ligases .</description>
  </primary_outcome>
  <number_of_groups>2</number_of_groups>
  <enrollment type="Anticipated">90</enrollment>
  <condition>Group 1: Trauma Operation for Otherwise Healthy Patients</condition>
  <condition>Group 2: Primary Tumors of Mesenchymal Origin</condition>
  <arm_group>
    <arm_group_label>The role of RING ubiquitin ligases in osteogenic progenitors</arm_group_label>
    <description>Bone marrow (BM) will be collected from healthy patients undergoing orthopedic surgery. BM will be collected from disposable tissue that is removed during the normal sequence of the surgery.</description>
  </arm_group>
  <arm_group>
    <arm_group_label>The role of RING ubiquitin ligases in musculoskeletal cancer</arm_group_label>
    <description>Collection of connective tissue from patients with tumors of musculoskeletal origin - a part of the resected tumor specimens.</description>
  </arm_group>
  <biospec_retention>Samples Without DNA</biospec_retention>
  <biospec_descr>
    <textblock>
      1. Bone marrow will be collected from othewise healthy patients undergoing orthopedic&#xD;
           surgery - arthroplasty for the treatment of degenerative joint disease or traumatised&#xD;
           patients. The bone marrow will be collected from disposable tissue that is removed&#xD;
           during the normal sequence of the surgery.&#xD;
&#xD;
        2. Collection of connective tissue from patients with malignancies of musculoskeletal&#xD;
           origin. The tissue that will be used is part of the resected tumor specimens. The tissue&#xD;
           will be used for:&#xD;
    </textblock>
  </biospec_descr>
  <eligibility>
    <study_pop>
      <textblock>
        1. Healthy patients undergoing orthopedic surgery - arthroplasty for the treatment of&#xD;
             degenerative joint disease or traumatised patients.&#xD;
&#xD;
          2. Patients with malignancies of musculoskeletal origin.&#xD;
      </textblock>
    </study_pop>
    <sampling_method>Non-Probability Sample</sampling_method>
    <criteria>
      <textblock>
        Group 1:&#xD;
&#xD;
          -  Inclusion Criteria:&#xD;
&#xD;
          -  Healthy patients undergoing orthopedic surgery - arthroplasty for the treatment of&#xD;
             degenerative joint disease or traumatised patients.&#xD;
&#xD;
          -  Exclusion Criteria:&#xD;
&#xD;
          -  Any malignancy in the past or active infection.&#xD;
&#xD;
        Group 2:&#xD;
&#xD;
          -  Inclusion Criteria:&#xD;
&#xD;
          -  patients with malignancies of musculoskeletal origin.&#xD;
&#xD;
          -  Exclusion Criteria:&#xD;
&#xD;
          -  Any malignancy of unknown origin or malignancy of multiple origins or active&#xD;
             infection.&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>18 Years</minimum_age>
    <maximum_age>N/A</maximum_age>
    <healthy_volunteers>No</healthy_volunteers>
  </eligibility>
  <location>
    <facility>
      <name>Rambam MC</name>
      <address>
        <city>Haifa</city>
        <country>Israel</country>
      </address>
    </facility>
    <status>Recruiting</status>
    <contact>
      <last_name>Rostislav Novak, MD/PhD</last_name>
      <phone>+972502061188</phone>
      <email>n_rosti@rambam.health.gov.il</email>
    </contact>
    <contact_backup>
      <last_name>Amir M Orian, MD/PhD</last_name>
      <phone>+97248295448/9</phone>
      <email>mdoryan@tx.technion.ac.il</email>
    </contact_backup>
    <investigator>
      <last_name>David Nikomarov, MD</last_name>
      <role>Principal Investigator</role>
    </investigator>
    <investigator>
      <last_name>Rostislav Novak, MD/PhD</last_name>
      <role>Sub-Investigator</role>
    </investigator>
    <investigator>
      <last_name>Amir Oryan (Orian), MD/PhD</last_name>
      <role>Sub-Investigator</role>
    </investigator>
  </location>
  <location_countries>
    <country>Israel</country>
  </location_countries>
  <verification_date>February 2015</verification_date>
  <study_first_submitted>July 29, 2014</study_first_submitted>
  <study_first_submitted_qc>September 30, 2014</study_first_submitted_qc>
  <study_first_posted type="Estimate">October 3, 2014</study_first_posted>
  <last_update_submitted>February 22, 2015</last_update_submitted>
  <last_update_submitted_qc>February 22, 2015</last_update_submitted_qc>
  <last_update_posted type="Estimate">February 24, 2015</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor</responsible_party_type>
  </responsible_party>
  <!-- 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/NCT02256241
org study id: 0180-14-RMB
nct id: NCT02256241
lead sponsor: 
collaborator: 
has dmc: No
Stage 1:

      Bone marrow will be collected from otherwise healthy patients undergoing orthopedic surgery -
      arthroplasty for the treatment of degenerative joint disease or traumatised patients. The
      bone marrow will be collected from disposable tissue that is removed during the normal
      sequence of the surgery. These cells will be used for:

      Establishment of the role of RING ubiquitin ligases in osteogenic progenitors proliferation
      and differentiation in culture.

      I. To determine the levels of RING ubiquitin ligases mRNA and protein in differentiating
      human mesenchymal precursor cells: To test the correlation between levels of RING ubiquitin
      ligases and degree of osteogenic differentiation, we will extract mesenchymal precursor cells
      from the collected tissue and test the mRNA protein level of RING ubiquitin ligases upon
      induction of differentiation. Specifically, we will compare the levels of RING ubiquitin
      ligases before and after the initiation of differentiational stimulus in time dependent
      manner by western blot and real time PCR analysis.

      II. To test the impact of specific RING ubiquitin ligases on differentiation of human
      mesenchymal precursor cells: to test the impact of RING ubiquitin ligases on the
      differentiation of mesenchymal precursor cell from the collected tissue by testing the
      expression of classical differentiational bone markers by flow cytometry (fibronectin, CD105)
      and by Alkaline phosphatase (ALP) activity assay. For this aim we developed both a
      constitutive and Dox-regulated conditional overexpression and shRNA lenti-viral systems that
      enables efficient modulation of these RING ubiquitin ligases level.

      III. Determine the role of RING ubiquitin ligases in proliferation and survival of human
      mesenchymal precursor cells: Via inhibition or overexpression of ligases in mesenchymal
      progenitor cells we will test the role of these ligases in proliferation and survival of
      mesenchymal precursors by using MTT assay, Propidion-Iodid (PI) and tunnel assays in flow
      cytometry analysis.

      Stage 2:

      Collection of connective tissue from patients with malignancies of musculoskeletal origin.
      The tissue that will be used is part of the resected tumor specimens. The tissue will be used
      for:

      The establishment of the role/s of RING ligases in musculoskeletal cancers using cell culture
      and in vivo activation. Test if the expression of selected positive candidates from stage 1
      correlates with cancer development and progression in human-derived samples Independently of
      our mechanistic experiment we aim to determine the relevance of these ligases to human
      musculoskeletal cancers. As the first step we will screen primary tumor biopsies at the
      protein level correlates with cancer grade and prognosis. Toward this aim we recently
      generated a highly specific several anti-monoclonal antibodies in our laboratory as well use
      comercial available antibodies .
Stage 1:

      Bone marrow will be collected from otherwise healthy patients undergoing orthopedic surgery -
      arthroplasty for the treatment of degenerative joint disease or traumatised patients. The
      bone marrow will be collected from disposable tissue that is removed during the normal
      sequence of the surgery. These cells will be used for:

      Establishment of the role of RING ubiquitin ligases in osteogenic progenitors proliferation
      and differentiation in culture.

      I. To determine the levels of RING ubiquitin ligases mRNA and protein in differentiating
      human mesenchymal precursor cells: To test the correlation between levels of RING ubiquitin
      ligases and degree of osteogenic differentiation, we will extract mesenchymal precursor cells
      from the collected tissue and test the mRNA protein level of RING ubiquitin ligases upon
      induction of differentiation. Specifically, we will compare the levels of RING ubiquitin
      ligases RING ubiquitin ligases before and after the initiation of differentiational stimulus
      in time dependent manner by western blot and real time PCR analysis.

      II. To test the impact of RING ubiquitin ligases on differentiation of human mesenchymal
      precursor cells: to test the impact of RING ubiquitin ligases on the differentiation of
      mesenchymal precursor cell from the collected tissue by testing the expression of classical
      differentiational bone markers by flow cytometry (fibronectin, CD105) and by Alkaline
      phosphatase (ALP) activity assay. For this aim we developed both a constitutive and
      Dox-regulated conditional RING ubiquitin ligases overexpression and shRNA lenti-viral systems
      that enables efficient modulation of RING ubiquitin ligases level.

      III. Determine the role of RING ubiquitin ligases in proliferation and survival of human
      mesenchymal precursor cells: We will test the role of their in proliferation and survival of
      mesenchymal precursors by using MTT assay, Propidion-Iodid (PI) and tunnel assays in flow
      cytometry analysis.

      Stage 2:

      Collection of connective tissue from patients with malignancies of musculoskeletal origin.
      The tissue that will be used is part of the resected tumor specimens. The tissue will be used
      for:

      The establishment of the role/s of RING ubiquitin ligases in musculoskeletal cancers using
      cell culture and in vivo.

      I. Test the role of our in proliferation and survival in musculoskeletal cancers: Using our
      lentiviral systems we will test the biological impact of inhibition and overexpression in
      musculoskeletal neoplastic cells.

      II. Test whether inhibition or overexpression of these ligases impacts the biology of
      musculoskeletal cancers.We will check if inhibition of these ligases in malignant cells of
      musculoskeletal origin by lenti-viral infections induces differentiation. Infected cells will
      be analyzed for expression of differentiational markers in flow cytometry and
      immunohistochemistry.

      III. Test if expression of RING ubiquitin ligases correlates with cancer development and
      progression in human-derived samples Independently of our mechanistic experiment we aim to
      determine the relevance of RING ubiquitin ligases to human musculoskeletal cancers.

      Patients consent will be signed after properly informed at time of pre-surgical preparation
      visit. In cases when pre-surgical visit is not held consent will be signed prior to the day
      of surgery.
observational model: Cohort
measure: Protein and mRNA levels of RING ubiquitin ligases during osteogenesis
time frame: 5 years
description: To determine the levels of RING ubiquitin ligases mRNA and protein in differentiating human mesenchymal precursor cells. We will extract mesenchymal precursor cells from the collected tissue and test the mRNA and protein levels of RING ubiquitin ligases upon induction of differentiation.
measure: Protein and mRNA levels of RING ubiquitin ligases in musculoskeletal tumors and its correlation to staging and grading
time frame: Time Frame: * 5 years
description: It is not clear if RING ubiquitin ligases protein levels are elevated in soft tissue-derived cancers (primary and metastatic tumors) and whether there is a correlation between RING ubiquitin ligases levels and staging and grading of soft tissue-derived cancers. For this reason, we will check whether RING ubiquitin ligases expression correlates with cancer development and progression in human-derived samples. We will determine the relevance of RING ubiquitin ligases to human musculoskeletal cancers. To do this we will screen primary tumor biopsies for RNA and protein levels of RING ubiquitin ligases .
arm group label: The role of RING ubiquitin ligases in osteogenic progenitors
description: Bone marrow (BM) will be collected from healthy patients undergoing orthopedic surgery. BM will be collected from disposable tissue that is removed during the normal sequence of the surgery.
arm group label: The role of RING ubiquitin ligases in musculoskeletal cancer
description: Collection of connective tissue from patients with tumors of musculoskeletal origin - a part of the resected tumor specimens.
1. Bone marrow will be collected from othewise healthy patients undergoing orthopedic
           surgery - arthroplasty for the treatment of degenerative joint disease or traumatised
           patients. The bone marrow will be collected from disposable tissue that is removed
           during the normal sequence of the surgery.

        2. Collection of connective tissue from patients with malignancies of musculoskeletal
           origin. The tissue that will be used is part of the resected tumor specimens. The tissue
           will be used for:
study pop: 
sampling method: Non-Probability Sample
criteria: 
gender: All
minimum age: 18 Years
maximum age: N/A
healthy volunteers: No
facility: 
status: Recruiting
contact: 
contact backup: 
investigator: 
investigator: 
investigator: 
country: Israel
responsible party type: Sponsor