<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/NCT04489251</url>
  </required_header>
  <id_info>
    <org_study_id>1492809</org_study_id>
    <nct_id>NCT04489251</nct_id>
  </id_info>
  <brief_title>Assessment of the TGF-beta Pathway and Micro-RNA in Pediatric Pulmonary Arterial Hypertension</brief_title>
  <official_title>Assessment of the TGF-beta Pathway and Micro-RNA in Pediatric Pulmonary Arterial Hypertension</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>Medical College of Wisconsin</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
  </sponsors>
  <source>Medical College of Wisconsin</source>
  <oversight_info>
    <has_dmc>No</has_dmc>
    <is_fda_regulated_drug>No</is_fda_regulated_drug>
    <is_fda_regulated_device>No</is_fda_regulated_device>
  </oversight_info>
  <brief_summary>
    <textblock>
      This is a prospective pilot study to assess the plasma levels of particular proteins involved&#xD;
      in the transforming growth factor beta (TGF-β) pathway and its down stream regulators, CHIP,&#xD;
      as well as micro RNA molecules in subjects with pulmonary arterial hypertension (PAH) and&#xD;
      compare them to control subjects without PAH to see if they can be used as a diagnostic or&#xD;
      prognostic marker of PAH and how this compares to other diagnostic biomarkers N-terminal&#xD;
      pro-natriuretic peptide (NT Pro-BNP) and C-reactive protein (CRP).&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      Aim 1: This study will correlate proteins in the TGF- β signaling pathway and micro RNA&#xD;
      levels with invasive (catheterization) and non-invasive (echocardiography) measurements of&#xD;
      pulmonary artery pressures to assess for the presence and severity of PAH and compare these&#xD;
      measurements to the established biomarkers of NT Pro BNP and CRP levels.&#xD;
&#xD;
      Hypothesis 1: Plasma levels of proteins of the TGF-β pathway; bone morphogenic protein (BMP)&#xD;
      2, 4, 6, 7, 9 and 10 along with activin A and TGF-β1 protein as well as CHIP&#xD;
      (carboxyl-terminus of Hsp70-intracting protein), an enzyme that regulates the activations and&#xD;
      exports of TGF- β to the nucleus will be significantly different in subjects with PH over&#xD;
      control subjects.&#xD;
&#xD;
      Hypothesis 2: Plasma levels of proteins in the TGF- β pathway; BMP 2, 4, 6, 7, 9 and 10 along&#xD;
      with activin A and TGF-β1 protein as well as CHIP will show better correlation with the&#xD;
      presence of PAH and its severity than NT-Pro BNP and CRP levels.&#xD;
&#xD;
      Hypothesis 3: The micro-RNA profiles in plasma will be significantly different in subjects&#xD;
      with PAHPH over control subjects.&#xD;
&#xD;
      Aim 2: To correlate protein/micro-RNA levels with clinical status in PAH subjects as assessed&#xD;
      by functional status, exercise testing, and PAH drug regimen to determine if they can&#xD;
      correlate with disease severity.&#xD;
&#xD;
      Hypothesis 1: Clinical findings in PAH patients will correlate with disease severity and&#xD;
      study proteins and micro-RNA levels better than established biomarkers.&#xD;
&#xD;
      Aim 3: To correlate evidence of genetic abnormalities through whole exome sequencing&#xD;
      especially in regions known or suspected to cause PAH (e.g. BMPR2, ENG, and ALK1 mutations),&#xD;
      within the TGF-β pathway or lung development with the tested protein/micro-RNA levels.&#xD;
&#xD;
      Hypothesis 1: Genetic evaluation of patients with PAH will show abnormalities within the&#xD;
      TGF-β pathway or lung development.&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Recruiting</overall_status>
  <start_date type="Actual">July 1, 2020</start_date>
  <completion_date type="Anticipated">January 1, 2023</completion_date>
  <primary_completion_date type="Anticipated">August 1, 2022</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>Prospective</time_perspective>
  </study_design_info>
  <primary_outcome>
    <measure>Plasma levels of BMP proteins of the TGF-β pathway</measure>
    <time_frame>One day</time_frame>
    <description>bone morphogenic protein (BMP) 2, 4, 6, 7, 9 and 10 levels will be obtained in PH subjects and will be compared to control subjects in effort to describe how those levels are different in subjects with PH over control subjects. All measured amounts of proteins will be in pg/ml.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Plasma Activin A and TGF-β1 level of the TGF-β pathway</measure>
    <time_frame>One day</time_frame>
    <description>Activin A and TGF-β1 plasma levels will be obtained in PH subjects and compared to control subjects in effort to describe how those levels are different in subjects with PH over control subjects. All measured amounts will be will be in pg/ml.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Plasma CHIP levels assessment in PH patients and control subjects.</measure>
    <time_frame>One day</time_frame>
    <description>CHIP (carboxyl-protein terminus of Hsp70-intracting protein) plasma levels will be obtained in PH subjects and compared to control subjects in effort to describe how those levels are different in subjects with PH over control subjects. All measured levels will be in pg/ml</description>
  </primary_outcome>
  <secondary_outcome>
    <measure>Plasma levels of proteins in the TGF- β pathway and PH disease severity</measure>
    <time_frame>One day</time_frame>
    <description>BMP 2, 4, 6, 7, 9 and 10 along with activin A and TGF-β1 protein as well as CHIP levels will be correlated with the presence of PAH and its severity and how that compares to the levels of NT-Pro BNP and CRP levels.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>MicroRNA levels in PH vs control subjects</measure>
    <time_frame>One day</time_frame>
    <description>The micro-RNA profiles in plasma will be compared between in subjects with PAH and control subjects.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Clinical findings correlation with study proteins/microRNA</measure>
    <time_frame>One day</time_frame>
    <description>Clinical findings in PAH patients will be correlated with disease severity and study protein and micro-RNA levels.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Whole exome sequence analysis in PH patients and protein/microRNA levels</measure>
    <time_frame>One day</time_frame>
    <description>To correlate evidence of genetic abnormalities through whole exome sequencing especially in regions known or suspected to cause PAH (e.g. BMPR2, ENG, and ALK1 mutations), within the TGF-β pathway or lung development with the tested protein/micro-RNA levels.</description>
  </secondary_outcome>
  <number_of_groups>2</number_of_groups>
  <enrollment type="Anticipated">40</enrollment>
  <condition>Pulmonary Arterial Hypertension</condition>
  <arm_group>
    <arm_group_label>PH subjects</arm_group_label>
    <description>Pediatric subjects ages 2-17 years&#xD;
Subjects undergoing a clinically indicated cardiac catheterization.&#xD;
Subjects with proven or being evaluated for pulmonary hypertension in WHO classification group 1 or 3†&#xD;
Subjects will be categorized as PAH subjects if they meet the hemodynamic criteria: pulmonary artery pressure &gt;20mmHg, pulmonary vascular resistance index &gt;3 Woods units*m2, and wedge pressures &lt;15mmHg.</description>
  </arm_group>
  <arm_group>
    <arm_group_label>Control subjects</arm_group_label>
    <description>Pediatric subjects ages 2-17 years&#xD;
Subjects undergoing a clinically indicated cardiac catheterization.&#xD;
Subjects can be categorized as control subjects if they do not have PH on catheterization and do not meet any exclusion criteria.</description>
  </arm_group>
  <intervention>
    <intervention_type>Diagnostic Test</intervention_type>
    <intervention_name>Protein Elisa analysis, microRNA analysis, whole exome sequencing</intervention_name>
    <description>ELISA Testing Procedure: The circulating levels of BMP 2, 4, 6, 7, 9 and 10, together with activin A and TGF-β as well as CHIP will be measured using streptavidin ELISA assays.&#xD;
Micro-RNA analysis: From 100µL of plasma, we will prepare small RNA libraries using TruSeq Small RNA Sample Preparation kit (Illumina Inc).&#xD;
TGF-B mutation evaluation: Blood from subjects with PH will also be analyzed for known genetic mutations in TGF-B pathway using whole exome sequencing.</description>
    <arm_group_label>Control subjects</arm_group_label>
    <arm_group_label>PH subjects</arm_group_label>
  </intervention>
  <biospec_retention>Samples With DNA</biospec_retention>
  <biospec_descr>
    <textblock>
      Serum is obtained from the pulmonary artery in patients undergoing a cardiac catheterization.&#xD;
    </textblock>
  </biospec_descr>
  <eligibility>
    <study_pop>
      <textblock>
        Subjects with previously confirmed or suspected to have pulmonary arterial hypertension&#xD;
        (PAH) that are undergoing a clinically indicated cardiac catheterization (for diagnostic&#xD;
        evaluation) ages 2-17 years are eligible for participation.&#xD;
&#xD;
        Control subjects undergoing a catheterization as part of an intervention such as atrial&#xD;
        septal defect closure or patent ductus arteriosus closure and have a routine right heart&#xD;
        catheterization as a part of that procedure and meet inclusion criteria.&#xD;
      </textblock>
    </study_pop>
    <sampling_method>Non-Probability Sample</sampling_method>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  Pediatric subjects ages 2-17 years&#xD;
&#xD;
          -  Subjects undergoing a clinically indicated cardiac catheterization.&#xD;
&#xD;
          -  Subjects with proven or being evaluated for pulmonary hypertension in WHO&#xD;
             classification group 1 or 3†&#xD;
&#xD;
          -  Subjects will be categorized as PAH subjects if they meet the hemodynamic criteria:&#xD;
             pulmonary artery pressure &gt;20mmHg, pulmonary vascular resistance index &gt;3 Woods&#xD;
             units*m2, and wedge pressures &lt;15mmHg.&#xD;
&#xD;
          -  Subjects can be categorized as control subjects if they do not have PH on&#xD;
             catheterization and do not meet any exclusion criteria.&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
        -&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>2 Years</minimum_age>
    <maximum_age>17 Years</maximum_age>
  </eligibility>
  <overall_official>
    <last_name>Edward C Kirkpatrick</last_name>
    <role>Principal Investigator</role>
    <affiliation>Children's Hospital and Health System Foundation, Wisconsin</affiliation>
  </overall_official>
  <overall_contact>
    <last_name>Edward C Kirkpatrick, DO</last_name>
    <phone>414-266-2380</phone>
    <email>ekirkpatrick@chw.org</email>
  </overall_contact>
  <overall_contact_backup>
    <last_name>Nicholas Peterson, BA</last_name>
    <phone>414-266-1753</phone>
    <email>NPeterson@chw.org</email>
  </overall_contact_backup>
  <location>
    <facility>
      <name>Children's Hospital of Wisconsin</name>
      <address>
        <city>Milwaukee</city>
        <state>Wisconsin</state>
        <zip>53226</zip>
        <country>United States</country>
      </address>
    </facility>
    <status>Recruiting</status>
    <contact>
      <last_name>Edward C Kirkpatrick, DO</last_name>
      <phone>414-266-2380</phone>
      <email>ekirkpatrick@chw.org</email>
    </contact>
    <contact_backup>
      <last_name>Nicholas Peterson, BA</last_name>
      <phone>414-266-1753</phone>
      <email>npeterson@chw.org</email>
    </contact_backup>
  </location>
  <location_countries>
    <country>United States</country>
  </location_countries>
  <verification_date>November 2021</verification_date>
  <study_first_submitted>July 21, 2020</study_first_submitted>
  <study_first_submitted_qc>July 23, 2020</study_first_submitted_qc>
  <study_first_posted type="Actual">July 28, 2020</study_first_posted>
  <last_update_submitted>November 12, 2021</last_update_submitted>
  <last_update_submitted_qc>November 12, 2021</last_update_submitted_qc>
  <last_update_posted type="Actual">November 22, 2021</last_update_posted>
  <responsible_party>
    <responsible_party_type>Principal Investigator</responsible_party_type>
    <investigator_affiliation>Medical College of Wisconsin</investigator_affiliation>
    <investigator_full_name>Edward Kirkpatrick</investigator_full_name>
    <investigator_title>Associate Professor</investigator_title>
  </responsible_party>
  <keyword>pulmonary arterial hypertension</keyword>
  <keyword>TGF-beta pathway</keyword>
  <keyword>microRNA</keyword>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Pulmonary Arterial Hypertension</mesh_term>
    <mesh_term>Familial Primary Pulmonary Hypertension</mesh_term>
    <mesh_term>Hypertension</mesh_term>
  </condition_browse>
  <patient_data>
    <sharing_ipd>No</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/NCT04489251
org study id: 1492809
nct id: NCT04489251
lead sponsor: 
has dmc: No
is fda regulated drug: No
is fda regulated device: No
This is a prospective pilot study to assess the plasma levels of particular proteins involved
      in the transforming growth factor beta (TGF-β) pathway and its down stream regulators, CHIP,
      as well as micro RNA molecules in subjects with pulmonary arterial hypertension (PAH) and
      compare them to control subjects without PAH to see if they can be used as a diagnostic or
      prognostic marker of PAH and how this compares to other diagnostic biomarkers N-terminal
      pro-natriuretic peptide (NT Pro-BNP) and C-reactive protein (CRP).
Aim 1: This study will correlate proteins in the TGF- β signaling pathway and micro RNA
      levels with invasive (catheterization) and non-invasive (echocardiography) measurements of
      pulmonary artery pressures to assess for the presence and severity of PAH and compare these
      measurements to the established biomarkers of NT Pro BNP and CRP levels.

      Hypothesis 1: Plasma levels of proteins of the TGF-β pathway; bone morphogenic protein (BMP)
      2, 4, 6, 7, 9 and 10 along with activin A and TGF-β1 protein as well as CHIP
      (carboxyl-terminus of Hsp70-intracting protein), an enzyme that regulates the activations and
      exports of TGF- β to the nucleus will be significantly different in subjects with PH over
      control subjects.

      Hypothesis 2: Plasma levels of proteins in the TGF- β pathway; BMP 2, 4, 6, 7, 9 and 10 along
      with activin A and TGF-β1 protein as well as CHIP will show better correlation with the
      presence of PAH and its severity than NT-Pro BNP and CRP levels.

      Hypothesis 3: The micro-RNA profiles in plasma will be significantly different in subjects
      with PAHPH over control subjects.

      Aim 2: To correlate protein/micro-RNA levels with clinical status in PAH subjects as assessed
      by functional status, exercise testing, and PAH drug regimen to determine if they can
      correlate with disease severity.

      Hypothesis 1: Clinical findings in PAH patients will correlate with disease severity and
      study proteins and micro-RNA levels better than established biomarkers.

      Aim 3: To correlate evidence of genetic abnormalities through whole exome sequencing
      especially in regions known or suspected to cause PAH (e.g. BMPR2, ENG, and ALK1 mutations),
      within the TGF-β pathway or lung development with the tested protein/micro-RNA levels.

      Hypothesis 1: Genetic evaluation of patients with PAH will show abnormalities within the
      TGF-β pathway or lung development.
observational model: Case-Control
time perspective: Prospective
measure: Plasma levels of BMP proteins of the TGF-β pathway
time frame: One day
description: bone morphogenic protein (BMP) 2, 4, 6, 7, 9 and 10 levels will be obtained in PH subjects and will be compared to control subjects in effort to describe how those levels are different in subjects with PH over control subjects. All measured amounts of proteins will be in pg/ml.
measure: Plasma Activin A and TGF-β1 level of the TGF-β pathway
time frame: One day
description: Activin A and TGF-β1 plasma levels will be obtained in PH subjects and compared to control subjects in effort to describe how those levels are different in subjects with PH over control subjects. All measured amounts will be will be in pg/ml.
measure: Plasma CHIP levels assessment in PH patients and control subjects.
time frame: One day
description: CHIP (carboxyl-protein terminus of Hsp70-intracting protein) plasma levels will be obtained in PH subjects and compared to control subjects in effort to describe how those levels are different in subjects with PH over control subjects. All measured levels will be in pg/ml
measure: Plasma levels of proteins in the TGF- β pathway and PH disease severity
time frame: One day
description: BMP 2, 4, 6, 7, 9 and 10 along with activin A and TGF-β1 protein as well as CHIP levels will be correlated with the presence of PAH and its severity and how that compares to the levels of NT-Pro BNP and CRP levels.
measure: MicroRNA levels in PH vs control subjects
time frame: One day
description: The micro-RNA profiles in plasma will be compared between in subjects with PAH and control subjects.
measure: Clinical findings correlation with study proteins/microRNA
time frame: One day
description: Clinical findings in PAH patients will be correlated with disease severity and study protein and micro-RNA levels.
measure: Whole exome sequence analysis in PH patients and protein/microRNA levels
time frame: One day
description: To correlate evidence of genetic abnormalities through whole exome sequencing especially in regions known or suspected to cause PAH (e.g. BMPR2, ENG, and ALK1 mutations), within the TGF-β pathway or lung development with the tested protein/micro-RNA levels.
arm group label: PH subjects
description: Pediatric subjects ages 2-17 years
Subjects undergoing a clinically indicated cardiac catheterization.
Subjects with proven or being evaluated for pulmonary hypertension in WHO classification group 1 or 3†
Subjects will be categorized as PAH subjects if they meet the hemodynamic criteria: pulmonary artery pressure >20mmHg, pulmonary vascular resistance index >3 Woods units*m2, and wedge pressures <15mmHg.
arm group label: Control subjects
description: Pediatric subjects ages 2-17 years
Subjects undergoing a clinically indicated cardiac catheterization.
Subjects can be categorized as control subjects if they do not have PH on catheterization and do not meet any exclusion criteria.
intervention type: Diagnostic Test
intervention name: Protein Elisa analysis, microRNA analysis, whole exome sequencing
description: ELISA Testing Procedure: The circulating levels of BMP 2, 4, 6, 7, 9 and 10, together with activin A and TGF-β as well as CHIP will be measured using streptavidin ELISA assays.
Micro-RNA analysis: From 100µL of plasma, we will prepare small RNA libraries using TruSeq Small RNA Sample Preparation kit (Illumina Inc).
TGF-B mutation evaluation: Blood from subjects with PH will also be analyzed for known genetic mutations in TGF-B pathway using whole exome sequencing.
arm group label: Control subjects
arm group label: PH subjects
Serum is obtained from the pulmonary artery in patients undergoing a cardiac catheterization.
study pop: 
sampling method: Non-Probability Sample
criteria: 
gender: All
minimum age: 2 Years
maximum age: 17 Years
last name: Edward C Kirkpatrick
role: Principal Investigator
affiliation: Children's Hospital and Health System Foundation, Wisconsin
last name: Edward C Kirkpatrick, DO
phone: 414-266-2380
email: ekirkpatrick@chw.org
last name: Nicholas Peterson, BA
phone: 414-266-1753
email: NPeterson@chw.org
facility: 
status: Recruiting
contact: 
contact backup: 
country: United States
responsible party type: Principal Investigator
investigator affiliation: Medical College of Wisconsin
investigator full name: Edward Kirkpatrick
investigator title: Associate Professor
mesh term: Pulmonary Arterial Hypertension
mesh term: Familial Primary Pulmonary Hypertension
mesh term: Hypertension
sharing ipd: No