<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/NCT03828422</url>
  </required_header>
  <id_info>
    <org_study_id>TP20180038</org_study_id>
    <nct_id>NCT03828422</nct_id>
  </id_info>
  <brief_title>Arterial Function and Atherosclerosis in Essential Thrombocythemia</brief_title>
  <official_title>Arterial Function and Atherosclerosis in Patients With JAK2 V167F Positive Essential Thrombocythemia</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>University Medical Centre Ljubljana</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
  </sponsors>
  <source>University Medical Centre Ljubljana</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>
      The aim of the study is to examine (a) whether patients with JAK2 V617F positive ET in&#xD;
      comparison to age-and sex-matched, apparently healthy control subjects show more advanced&#xD;
      progression of arterial stiffness, pulse-wave velocity and coronary calcium score in a 4 year&#xD;
      observation period, and (b) whether the burden of JAK2 V617F mutation correlates with the&#xD;
      measured vascular parameters. All subjects will be examined twice. The first visit already&#xD;
      took place between the years 2014 - 2015 and the second visit will take place between&#xD;
      2018-2019. All participants will have signed their informed consent before entering the&#xD;
      study. Each visit will consist of completing a structured questionnaire (on personal and&#xD;
      family medical history, risk factors for CVD and medication), physical examination, donating&#xD;
      a blood sample for laboratory tests and undergoing carotid ultrasound and coronary calcium&#xD;
      measurement oft the extent of coronary artery calcification. At the first and the second&#xD;
      examination the JAK2 V617F allele burden, i.e. the percentage of mutated alleles, will be&#xD;
      determined from genomic DNA in peripheral blood.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      1. Patients and control subjects&#xD;
&#xD;
           Patients are selected from the database of the Department of Haematology at University&#xD;
           Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET&#xD;
           between 2011 and 2014. There were 124 JAK2 positive ET patients, among those 61 did not&#xD;
           have a personal history of clinically manifest atherosclerotic vascular disease&#xD;
           (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease,&#xD;
           transient ischemic attack or ischemic stroke). 40 patients (14 male and 26 female) with&#xD;
           JAK2 V617F positive ET without clinically apparent cardiovascular disease signed the&#xD;
           informed consent and were enrolled in the study in 2014 - 2015 for the first examination&#xD;
           and 36 (12 male and 24 female) of them are expected to participate also in 2018-19.&#xD;
&#xD;
           The control group is selected among healthy employees of the University Medical Centre&#xD;
           Ljubljana and their relatives. It is matched with the patient group for age and sex&#xD;
           distribution and classical risk factors for cardiovascular disease. 42 individuals (16&#xD;
           male and 26 female) participated in the first examination and at least 38 (14 male and&#xD;
           24 female) subjects are expected to paticipate in the second examination.&#xD;
&#xD;
        2. Baseline measurements&#xD;
&#xD;
           Each participant will complete a questionnaire about family history of cardiovascular&#xD;
           diseases, personal medical history, smoking status and medications. A physical&#xD;
           examination will be performed, including measurements of height, weight, waist&#xD;
           circumference, systolic and diastolic blood pressure. Blood will be sampled for&#xD;
           laboratory analysis of complete blood cell count, electrolytes, serum lipids, liver&#xD;
           function tests, urea, creatinine, and inflammatory markers.&#xD;
&#xD;
           The 10-year risk of coronary heart disease (CHD) and general cardiovascular disease&#xD;
           (CVD) will be calculated using the Framingham risk equation.&#xD;
&#xD;
        3. JAK2 V617F/G1849T allele burden&#xD;
&#xD;
           The ipsogen JAK2 MutaQuant Kit, Qiagen (ZDA) (Ref: No. 673523) will be used for the&#xD;
           detection and quantification of JAK2 V617F/G1849T allele in genomic DNA extracted from&#xD;
           peripheral blood of patients and also control subjects. A SNP specific primer&#xD;
           selectively amplifies the JAK2 V617F allele which is detected with a real-time qPCR&#xD;
           instrument that quantifies the PCR products. The JAK2 V617F allele burden will be&#xD;
           calculated and expressed as the percentage of JAK2 V617F mutated alleles throughout the&#xD;
           whole JAK2 record. Analysis will be done done in laboratory of the Department of&#xD;
           Haematology at University Medical Centre Ljubljana for all samples.&#xD;
&#xD;
        4. Carotid Artery Ultrasound Examination&#xD;
&#xD;
           For examination of the extracranial carotid arteries an ultrasound machine Aloka&#xD;
           Prosound α7 (Hitachi Aloka Medical, Ltd., Japan) will be used with a linear vascular&#xD;
           probe working at a frequency of 5-13 MHz. The common, internal and external carotid&#xD;
           arteries on both sided will be examined. At each examination, the measurements will be&#xD;
           done twice and their average values will be calculated. The ultrasound examination will&#xD;
           be done by the same ultrasonographer at both visits of all participants.&#xD;
&#xD;
           The intima-media thickness will be measured 2 cm proximal to the bulb of common carotid&#xD;
           artery on both sides. Screening of the extracranial carotid arteries for atherosclerotic&#xD;
           plaques will be performed, with a plaque definition of a focal lesion, exceeding the&#xD;
           intima-media thickness for at least 50% or reaching an absolute thickness of at least&#xD;
           1.5 mm in two orthogonal projections. Scoring of atherosclerotic plaques is done&#xD;
           according to the methodology from the Rotterdam Study. The presence of at least one&#xD;
           plaque in each segment of the extracranial carotid arterial bed, (divided into the&#xD;
           common carotid artery and the bulb, the internal carotid artery and the external carotid&#xD;
           artery) on either sides is scored 1 point. Thus, the carotid plaque score ranges from 0&#xD;
           (absence of plaques) to 6 (plaques present in all segments on both sides).&#xD;
&#xD;
           Echo-tracking of the common carotid artery will be used to assess arterial stiffness&#xD;
           expressed by the β-stiffness index and the pulse wave velocity. Measurements will be&#xD;
           done at the common carotid artery 2 cm proximal to the bulb on both sides. The&#xD;
           β-stiffness index will be calculated as: β = ln (P_max / P_min) / [(D_max - D_min /&#xD;
           D_min)], where P_max = the systolic blood pressure, P_min = the diastolic pressure;&#xD;
           D_max = the maximum arterial diameter and D_min = the minimal arterial diameter. Pulse&#xD;
           wave velocity (PWV) will be calculated as: PWV = √ ((β x P_min) / 2ρ)); ρ = 1050 kg/m³.&#xD;
&#xD;
        5. Assessment of Endothelial Function of the Digital Arteries&#xD;
&#xD;
           Endothelial function of the digital arteries will be measured by digital plethymography&#xD;
           with EndoPat 2000, Itamar Medical REF, Caesarea, Israel, Software Version 3.3.x and&#xD;
           expressed as the Reactive Hyperaemia Index (RHI) and the Augmentation Index (AI).&#xD;
           Changes in arterial tone are elicited by creating a downstream hyperemic response&#xD;
           induced by a standard 5-minute occlusion of the brachial artery (using a blood pressure&#xD;
           cuff, inflated to 60 mmHg above the arterial blood pressure). When the cuff is released,&#xD;
           the surge of blood flow causes endothelium-dependent flow mediated dilatation (FMD)&#xD;
           which is manifested as reactive hyperemia. A post-occlusion to pre-occlusion ratio is&#xD;
           calculated by EndoPAT software and the and A) are determined. The results are normalized&#xD;
           to a heart rate of 75/min.&#xD;
&#xD;
        6. Measurement of the Coronary Artery Calcium Burden&#xD;
&#xD;
           The calcium burden of coronary arteries is measured with a Biograph M 128-row PET-CT&#xD;
           scanner (Siemens, Erlangen, Germany). Scanning is done from the base to the apex of the&#xD;
           heart. A non-contrast protocol with sequential prospective ECG triggering is used. The&#xD;
           rotation time is 0.33 sec, with a tube voltage of 120 kV, CARE Dose 4D and slice&#xD;
           thickness 3 mm, with no slice overlap. Post-processing is done on the Syngo Leonardo&#xD;
           workstation. The coronary calcium burden is expressed as the Agatston score. Evaluation&#xD;
           of the dataset of every study subject is done three times and the average value is used&#xD;
           for further analysis.&#xD;
&#xD;
        7. Statistical analysis&#xD;
&#xD;
      All sets of data will be tested for normality of distribution using the normal-quintile plot,&#xD;
      calculating the correlation coefficient and checking it for the critical value that would&#xD;
      warrant rejection of normal distribution with an α-error probability of 0.05.&#xD;
&#xD;
      Normally distributed data will be presented as mean and standard deviation, while&#xD;
      non-normally distributed data will be presented as median and range between the 1st and 3rd&#xD;
      quartile.&#xD;
&#xD;
      Differences between subjects with ET and control subjects at the first and second examination&#xD;
      will be tested by the chi-square test for discrete variables, for normally distributed&#xD;
      continuous variables by the paired Student's t-test for independent samples, and for&#xD;
      non-normally distributed continuous variables by the Mann-Whitney test for independent&#xD;
      samples. P-values of &lt; 0.05 will be considered significant with a correction for multiple&#xD;
      comparisons.&#xD;
&#xD;
      Changes in vascular function and morphology in the 4-year observation period will be compared&#xD;
      between the two groups by the log rank test. P-values of &lt; 0.05 will be considered&#xD;
      significant.&#xD;
&#xD;
      The association between the parameters of vascular function / morphology and the JAK2 V617F&#xD;
      allele burden will be assessed by the Pearson correlation coefficient.&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Completed</overall_status>
  <start_date type="Actual">January 2014</start_date>
  <completion_date type="Actual">August 5, 2021</completion_date>
  <primary_completion_date type="Actual">August 2021</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>Carotid Artery Stiffness (Expressed by Beta-stiffness Index ) in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period (From Baseline to Year 4).</measure>
    <time_frame>From Baseline (in 2014-2015) to Year 4 (in 2018-2019)</time_frame>
    <description>4-year change carotid artery in Beta-stiffness Index measured in JAK2 V617F positive ET patients in comparison to healthy control subjects. All measurements were performed by echo-tracking ultrasound of the common carotid artery.&#xD;
Echo-tracking of the common carotid artery wall was used to assess arterial stiffness expressed by the β-stiffness index. Measurement swere done at the common carotid artery 2 cm proximal to the bulb on both sides. The β-stiffness index was calculated as: β = ln (P_max / P_min) / [(D_max - D_min / D_min)] (P_max = the systolic blood pressure, P_min = the diastolic pressure; D_max = the maximum arterial diameter and D_min = the minimal arterial diameter). In the adult population, the values of β range from 4 - 10 and increase with age. A larger value of β means increased arterial stiffness, i.e., a worse outcome.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Carotid Artery Stiffness (Expressed by the Pulse Wave Velocity) in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period (From Baseline to Year 4).</measure>
    <time_frame>From Baseline (in 2014-2015) to Year 4 (in 2018-2019)</time_frame>
    <description>4-year change in carotid artery pulse wave velocity (PWV) measured in JAK2 V617F positive ET patients in comparison to healthy control subjects. All measurements were performed by echo-tracking ultrasound of the common carotid artery.&#xD;
Measurements were done at the common carotid artery 2 cm proximal to the bulb on both sides. PWV was calculated from the beta stiffness index (β, for description see above), as follows:&#xD;
PWV = √ ((β x P_min) / 2ρ)); ρ = 1050 kg/m³. PWV increases with arterial stiffness, a larger value means a worse outcome. PWV in adults ranges from 3 m/s to 12 m/s; values are strongly age dependent.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Change of Carotid Artery Plaque Score in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period.</measure>
    <time_frame>From Baseline (in 2014-2015) to Year 4 (in 2018-2019)</time_frame>
    <description>Change in carotid plaque score (i.e., the number of carotid artery segments with detected plaques, reported on a scale from 0-6) in a 4 year observation period.&#xD;
A carotid plaque score 0 means that the common carotid, internal carotid and exteranal carotid artery on both sides are free from plaques. A score of 6 means that the common carotid, internal carotid and external carotid artery on both sides have plaques detected by ultrasound.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Change of Coronary Calcium Burden in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period.</measure>
    <time_frame>From Baseline (in 2014-2015) to Year 4 (in 2018-2019)</time_frame>
    <description>Change in the Agatston coronary artery calcium score in a 4 year observation period.&#xD;
Agatston score denotes the number of coronary artery calcifications weighted for the intensity of calcification.&#xD;
Grading of coronary artery disease (CAD) based od Agatston score goes as follows: no CAD = score 0, minimal CAD = score 1-10, mild CAD = score 11-100, moderate CAD = score 101-400, svere CAD = score &gt; 400.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Change of Digital Endothelial Function, Expressed as Reactive Hyperemia Index and Augmentation Index (AI), in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period.</measure>
    <time_frame>From Baseline (in 2014-2015) to Year 4 (in 2018-2019)</time_frame>
    <description>4-year change in Reactive Hyperemia Index (RHI) and 4-year change Augmentation Index (AI).&#xD;
RHI was calculated by the formula:&#xD;
RHI = (A/B) / (C/D) where A is the post-occlusion pulse wave amplitude (PWA) of the occluded hand, B the baseline PWA of the occluded hand, C the post-occlusion PWA of the contralateral hand, and D the baseline PWA of the contralateral hand. A higher RHI means better endothelial function (= good outcome). RHI in patients ranges from1-3. Arbitrarily, normal RHI is defined as =&gt; 1.67, and abnormal RHI as RHI =&lt; 1.66.&#xD;
The AI was determined from the shape of the arterial pulse wave by the EndoPAT 2000 software which distinguished between the primary pulse wave (P1) and the reflected pulse wave (P2) by the formula: AI = ((P2-P1)/P1) x 100. The results were normalized to a heart rate of 75/min. A higher AI means greater arterial stiffness (= bad outcome). AI &gt;0.4 is abnormal.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>The JAK2 V617F Mutation Burden in Patients With JAK2 V617F Positive ET.</measure>
    <time_frame>At baseline (in 2014- 2015) and at the second visit (in 2018-2019)</time_frame>
    <description>Change in JAK2 V617F mutation burden in the 4-year observation period. Ipsogen JAK2 MutaQuant Kit, Qiagen (USA) was used for the detection of JAK2 V617F mutation and quantification of its burden in genomic DNA, which was extracted from granulocytes in peripheral blood. Real-time quantitative polymerase chain reaction (qPCR) was used with double-dye oligonucleotide hydrolysis principle. The JAK2 V617F burden was defined as the percentage of JAK2 V617F mutated alleles in total genomic DNA.</description>
  </primary_outcome>
  <number_of_groups>2</number_of_groups>
  <enrollment type="Actual">82</enrollment>
  <condition>Atherosclerosis</condition>
  <arm_group>
    <arm_group_label>patients with essential thrombocythemia</arm_group_label>
    <description>essential thrombocythemia with JAK2 V617F positive mutation&#xD;
patients from the Department of Haematology at University Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET between 2011 and 2014&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
examined twice, the first time in the years 2014-2015 and for the second time in the years 2018-2019&#xD;
blood for laboratory tests&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning</description>
  </arm_group>
  <arm_group>
    <arm_group_label>control group</arm_group_label>
    <description>the control group is selected among healthy employees of the University Medical Centre Ljubljana and their relatives&#xD;
they are matched with the patient group for age and sex distribution and classical risk factors for cardiovascular disease&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
blood for laboratory tests&#xD;
examined twice, the first time in the years 2014-2015 and for the second time the in years 2018-2019&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning</description>
  </arm_group>
  <intervention>
    <intervention_type>Diagnostic Test</intervention_type>
    <intervention_name>imaging</intervention_name>
    <arm_group_label>control group</arm_group_label>
    <arm_group_label>patients with essential thrombocythemia</arm_group_label>
    <other_name>EndoPat plethysmography</other_name>
    <other_name>coronary artery calcium scanning</other_name>
    <other_name>blood laboratory tests</other_name>
    <other_name>ultrasound examination</other_name>
  </intervention>
  <biospec_retention>Samples With DNA</biospec_retention>
  <biospec_descr>
    <textblock>
      The ipsogen JAK2 MutaQuant Kit, Qiagen (ZDA) (Ref: No. 673523) will be used for the detection&#xD;
      and quantification of JAK2 V617F/G1849T allele in genomic DNA extracted from peripheral blood&#xD;
      of patients and also control subjects. A SNP specific primer that selectively amplifies JAK2&#xD;
      V617F allele will be used with a real-time qPCR instrument which accurately quantifies the&#xD;
      PCR products.&#xD;
    </textblock>
  </biospec_descr>
  <eligibility>
    <study_pop>
      <textblock>
        Expected patient group: 40 patients (14 male and 26 female) with JAK2 V617F positive ET&#xD;
        without clinically apparent cardiovascular disease. Average age 55,05 (13,48); ITM 25,22&#xD;
        (3,65); systolic blood pressure 138 (124-150) mmHg; diastolic blood pressure 81 (74-85)&#xD;
        mmHg; Framingham risk score for CHD 6,84%.&#xD;
&#xD;
        Expected control group: 42 individuals (16 male and 26 female) who were age and sex matched&#xD;
        with the patient group. Average age 59,16 (12,03); ITM 27,27 (4,64); systolic blood&#xD;
        pressure 132 (125-141) mmHg; diastolic blood pressure 81 (76-86) mmHg; Framingham risk&#xD;
        score for CHD 7,20%.&#xD;
      </textblock>
    </study_pop>
    <sampling_method>Probability Sample</sampling_method>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  patients with JAK2 V617F positive essential thrombocythemia&#xD;
&#xD;
          -  age-and sex-matched apparently healthy control subjects&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  personal history of any atherosclerotic vascular disease (myocardial infarction,&#xD;
             angina pectoris, peripheral arterial disease, aortic disease, transient ischemic&#xD;
             attack or ischemic stroke)&#xD;
&#xD;
          -  chronic kidney disease stage 3 and above&#xD;
&#xD;
          -  known cancer&#xD;
&#xD;
          -  chronic inflammatory disease&#xD;
&#xD;
          -  autoimmune disease&#xD;
&#xD;
          -  pregnancy&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>18 Years</minimum_age>
    <maximum_age>N/A</maximum_age>
    <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Ales Blinc, MD</last_name>
    <role>Study Director</role>
    <affiliation>University Medical Centre Ljubljana</affiliation>
  </overall_official>
  <results_reference>
    <citation>Vrtovec M, Anzic A, Zupan IP, Zaletel K, Blinc A. Carotid Artery Stiffness, Digital Endothelial Function, and Coronary Calcium in Patients with Essential Thrombocytosis, Free of Overt Atherosclerotic Disease. Radiol Oncol. 2017 Jan 14;51(2):203-210. doi: 10.1515/raon-2017-0006. eCollection 2017 Jun.</citation>
    <PMID>28740456</PMID>
  </results_reference>
  <results_reference>
    <citation>Anzic Drofenik A, Vrtovec M, Bozic Mijovski M, Sever M, Preloznik Zupan I, Kejzar N, Blinc A. Progression of coronary calcium burden and carotid stiffness in patients with essential thrombocythemia associated with JAK2 V617F mutation. Atherosclerosis. 2020 Mar;296:25-31. doi: 10.1016/j.atherosclerosis.2020.01.001. Epub 2020 Jan 11.</citation>
    <PMID>32005002</PMID>
  </results_reference>
  <verification_date>October 2021</verification_date>
  <study_first_submitted>January 23, 2019</study_first_submitted>
  <study_first_submitted_qc>January 30, 2019</study_first_submitted_qc>
  <study_first_posted type="Actual">February 4, 2019</study_first_posted>
  <results_first_submitted>March 26, 2020</results_first_submitted>
  <results_first_submitted_qc>October 6, 2021</results_first_submitted_qc>
  <results_first_posted type="Actual">November 4, 2021</results_first_posted>
  <last_update_submitted>October 6, 2021</last_update_submitted>
  <last_update_submitted_qc>October 6, 2021</last_update_submitted_qc>
  <last_update_posted type="Actual">November 4, 2021</last_update_posted>
  <responsible_party>
    <responsible_party_type>Principal Investigator</responsible_party_type>
    <investigator_affiliation>University Medical Centre Ljubljana</investigator_affiliation>
    <investigator_full_name>Ales Blinc</investigator_full_name>
    <investigator_title>Ales Blinc, MD, PhD, Head, Dept of Vascular Diseases</investigator_title>
  </responsible_party>
  <keyword>essential thrombocythaemia</keyword>
  <keyword>JAK2 V617F mutation burden</keyword>
  <keyword>preclinical atherosclerosis</keyword>
  <keyword>arterial stiffness</keyword>
  <keyword>endothelial function</keyword>
  <keyword>coronary artery calcium score</keyword>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Atherosclerosis</mesh_term>
    <mesh_term>Thrombocytosis</mesh_term>
    <mesh_term>Thrombocythemia, Essential</mesh_term>
  </condition_browse>
  <intervention_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Calcium</mesh_term>
  </intervention_browse>
  <patient_data>
    <sharing_ipd>Undecided</sharing_ipd>
  </patient_data>
  <provided_document_section>
    <provided_document>
      <document_type>Study Protocol and Statistical Analysis Plan</document_type>
      <document_has_protocol>Yes</document_has_protocol>
      <document_has_icf>No</document_has_icf>
      <document_has_sap>Yes</document_has_sap>
      <document_date>January 26, 2019</document_date>
      <document_url>https://ClinicalTrials.gov/ProvidedDocs/22/NCT03828422/Prot_SAP_000.pdf</document_url>
    </provided_document>
  </provided_document_section>

  <clinical_results>

    <participant_flow>
      <recruitment_details>a single tertiary medical centre</recruitment_details>
      <pre_assignment_details>had documented or clinically suspected atherosclerotic vascular disease</pre_assignment_details>
      <group_list>
        <group group_id="P1">
          <title>Patients With Essential Thrombocythemia</title>
          <description>essential thrombocythemia with JAK2 V617F positive mutation&#xD;
patients from the Department of Haematology at University Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET between 2011 and 2014&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
examined twice, the first time in the years 2014-2015 and for the second time in the years 2018-2019&#xD;
blood for laboratory tests&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
        </group>
        <group group_id="P2">
          <title>Control Group</title>
          <description>the control group is selected among healthy employees of the University Medical Centre Ljubljana and their relatives&#xD;
they are matched with the patient group for age and sex distribution and classical risk factors for cardiovascular disease&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
blood for laboratory tests&#xD;
examined twice, the first time in the years 2014-2015 and for the second time the in years 2018-2019&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
        </group>
      </group_list>
      <period_list>
        <period>
          <title>Overall Study</title>
          <milestone_list>
            <milestone>
              <title>STARTED</title>
              <participants_list>
                <participants group_id="P1" count="40"/>
                <participants group_id="P2" count="42"/>
              </participants_list>
            </milestone>
            <milestone>
              <title>COMPLETED</title>
              <participants_list>
                <participants group_id="P1" count="36"/>
                <participants group_id="P2" count="38"/>
              </participants_list>
            </milestone>
            <milestone>
              <title>NOT COMPLETED</title>
              <participants_list>
                <participants group_id="P1" count="4"/>
                <participants group_id="P2" count="4"/>
              </participants_list>
            </milestone>
          </milestone_list>
        </period>
      </period_list>
    </participant_flow>

    <baseline>
      <population>4 participants did not particiapete at the second visit</population>
      <group_list>
        <group group_id="B1">
          <title>Patients With Essential Thrombocythemia</title>
          <description>essential thrombocythemia with JAK2 V617F positive mutation&#xD;
patients from the Department of Haematology at University Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET between 2011 and 2014&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
examined twice, the first time in the years 2014-2015 and for the second time in the years 2018-2019&#xD;
blood for laboratory tests&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
        </group>
        <group group_id="B2">
          <title>Control Group</title>
          <description>the control group is selected among healthy employees of the University Medical Centre Ljubljana and their relatives&#xD;
they are matched with the patient group for age and sex distribution and classical risk factors for cardiovascular disease&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
blood for laboratory tests&#xD;
examined twice, the first time in the years 2014-2015 and for the second time the in years 2018-2019&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
        </group>
        <group group_id="B3">
          <title>Total</title>
          <description>Total of all reporting groups</description>
        </group>
      </group_list>
      <analyzed_list>
        <analyzed>
          <units>Participants</units>
          <scope>Overall</scope>
          <count_list>
            <count group_id="B1" value="36"/>
            <count group_id="B2" value="38"/>
            <count group_id="B3" value="74"/>
          </count_list>
        </analyzed>
      </analyzed_list>
      <measure_list>
        <measure>
          <title>Age</title>
          <units>years</units>
          <param>Mean</param>
          <dispersion>Standard Deviation</dispersion>
          <class_list>
            <class>
              <analyzed_list>
                <analyzed>
                  <units>Participants</units>
                  <scope>Class</scope>
                  <count_list>
                    <count group_id="B1" value="36"/>
                    <count group_id="B2" value="38"/>
                    <count group_id="B3" value="74"/>
                  </count_list>
                </analyzed>
              </analyzed_list>
              <category_list>
                <category>
                  <measurement_list>
                    <measurement group_id="B1" value="55" spread="13"/>
                    <measurement group_id="B2" value="59" spread="12"/>
                    <measurement group_id="B3" value="57" spread="13"/>
                  </measurement_list>
                </category>
              </category_list>
            </class>
          </class_list>
        </measure>
        <measure>
          <title>Sex: Female, Male</title>
          <units>Participants</units>
          <param>Count of Participants</param>
          <class_list>
            <class>
              <analyzed_list>
                <analyzed>
                  <units>Participants</units>
                  <scope>Class</scope>
                  <count_list>
                    <count group_id="B1" value="36"/>
                    <count group_id="B2" value="38"/>
                    <count group_id="B3" value="74"/>
                  </count_list>
                </analyzed>
              </analyzed_list>
              <category_list>
                <category>
                  <title>Female</title>
                  <measurement_list>
                    <measurement group_id="B1" value="24"/>
                    <measurement group_id="B2" value="24"/>
                    <measurement group_id="B3" value="48"/>
                  </measurement_list>
                </category>
                <category>
                  <title>Male</title>
                  <measurement_list>
                    <measurement group_id="B1" value="12"/>
                    <measurement group_id="B2" value="14"/>
                    <measurement group_id="B3" value="26"/>
                  </measurement_list>
                </category>
              </category_list>
            </class>
          </class_list>
        </measure>
        <measure>
          <title>Race and Ethnicity Not Collected</title>
          <population>Race and Ethnicity were not collected from any participant.</population>
          <units>Participants</units>
          <param>Count of Participants</param>
          <class_list>
            <class>
              <analyzed_list>
                <analyzed>
                  <units>Participants</units>
                  <scope>Class</scope>
                  <count_list>
                    <count group_id="B1" value="0"/>
                    <count group_id="B2" value="0"/>
                    <count group_id="B3" value="0"/>
                  </count_list>
                </analyzed>
              </analyzed_list>
              <category_list>
                <category>
                  <measurement_list>
                    <measurement group_id="B3" value="0"/>
                  </measurement_list>
                </category>
              </category_list>
            </class>
          </class_list>
        </measure>
        <measure>
          <title>Region of Enrollment</title>
          <units>participants</units>
          <param>Number</param>
          <class_list>
            <class>
              <title>Slovenia</title>
              <analyzed_list>
                <analyzed>
                  <units>Participants</units>
                  <scope>Class</scope>
                  <count_list>
                    <count group_id="B1" value="36"/>
                    <count group_id="B2" value="38"/>
                    <count group_id="B3" value="74"/>
                  </count_list>
                </analyzed>
              </analyzed_list>
              <category_list>
                <category>
                  <measurement_list>
                    <measurement group_id="B1" value="36"/>
                    <measurement group_id="B2" value="38"/>
                    <measurement group_id="B3" value="74"/>
                  </measurement_list>
                </category>
              </category_list>
            </class>
          </class_list>
        </measure>
      </measure_list>
    </baseline>

    <outcome_list>
      <outcome>
        <type>Primary</type>
        <title>Carotid Artery Stiffness (Expressed by Beta-stiffness Index ) in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period (From Baseline to Year 4).</title>
        <description>4-year change carotid artery in Beta-stiffness Index measured in JAK2 V617F positive ET patients in comparison to healthy control subjects. All measurements were performed by echo-tracking ultrasound of the common carotid artery.&#xD;
Echo-tracking of the common carotid artery wall was used to assess arterial stiffness expressed by the β-stiffness index. Measurement swere done at the common carotid artery 2 cm proximal to the bulb on both sides. The β-stiffness index was calculated as: β = ln (P_max / P_min) / [(D_max - D_min / D_min)] (P_max = the systolic blood pressure, P_min = the diastolic pressure; D_max = the maximum arterial diameter and D_min = the minimal arterial diameter). In the adult population, the values of β range from 4 - 10 and increase with age. A larger value of β means increased arterial stiffness, i.e., a worse outcome.</description>
        <time_frame>From Baseline (in 2014-2015) to Year 4 (in 2018-2019)</time_frame>
        <group_list>
          <group group_id="O1">
            <title>Patients With Essential Thrombocythemia</title>
            <description>essential thrombocythemia with JAK2 V617F positive mutation&#xD;
patients from the Department of Haematology at University Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET between 2011 and 2014&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
examined twice, the first time in the years 2014-2015 and for the second time in the years 2018-2019&#xD;
blood for laboratory tests&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
          </group>
          <group group_id="O2">
            <title>Control Group</title>
            <description>the control group is selected among healthy employees of the University Medical Centre Ljubljana and their relatives&#xD;
they are matched with the patient group for age and sex distribution and classical risk factors for cardiovascular disease&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
blood for laboratory tests&#xD;
examined twice, the first time in the years 2014-2015 and for the second time the in years 2018-2019&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
          </group>
        </group_list>
        <measure>
          <title>Carotid Artery Stiffness (Expressed by Beta-stiffness Index ) in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period (From Baseline to Year 4).</title>
          <description>4-year change carotid artery in Beta-stiffness Index measured in JAK2 V617F positive ET patients in comparison to healthy control subjects. All measurements were performed by echo-tracking ultrasound of the common carotid artery.&#xD;
Echo-tracking of the common carotid artery wall was used to assess arterial stiffness expressed by the β-stiffness index. Measurement swere done at the common carotid artery 2 cm proximal to the bulb on both sides. The β-stiffness index was calculated as: β = ln (P_max / P_min) / [(D_max - D_min / D_min)] (P_max = the systolic blood pressure, P_min = the diastolic pressure; D_max = the maximum arterial diameter and D_min = the minimal arterial diameter). In the adult population, the values of β range from 4 - 10 and increase with age. A larger value of β means increased arterial stiffness, i.e., a worse outcome.</description>
          <units>index</units>
          <param>Mean</param>
          <dispersion>Standard Deviation</dispersion>
          <analyzed_list>
            <analyzed>
              <units>Participants</units>
              <scope>Measure</scope>
              <count_list>
                <count group_id="O1" value="36"/>
                <count group_id="O2" value="38"/>
              </count_list>
            </analyzed>
          </analyzed_list>
          <class_list>
            <class>
              <category_list>
                <category>
                  <measurement_list>
                    <measurement group_id="O1" value="1.95" spread="2.18"/>
                    <measurement group_id="O2" value="0.22" spread="1.99"/>
                  </measurement_list>
                </category>
              </category_list>
            </class>
          </class_list>
        </measure>
      </outcome>
      <outcome>
        <type>Primary</type>
        <title>Carotid Artery Stiffness (Expressed by the Pulse Wave Velocity) in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period (From Baseline to Year 4).</title>
        <description>4-year change in carotid artery pulse wave velocity (PWV) measured in JAK2 V617F positive ET patients in comparison to healthy control subjects. All measurements were performed by echo-tracking ultrasound of the common carotid artery.&#xD;
Measurements were done at the common carotid artery 2 cm proximal to the bulb on both sides. PWV was calculated from the beta stiffness index (β, for description see above), as follows:&#xD;
PWV = √ ((β x P_min) / 2ρ)); ρ = 1050 kg/m³. PWV increases with arterial stiffness, a larger value means a worse outcome. PWV in adults ranges from 3 m/s to 12 m/s; values are strongly age dependent.</description>
        <time_frame>From Baseline (in 2014-2015) to Year 4 (in 2018-2019)</time_frame>
        <group_list>
          <group group_id="O1">
            <title>Patients With Essential Thrombocythemia</title>
            <description>essential thrombocythemia with JAK2 V617F positive mutation&#xD;
patients from the Department of Haematology at University Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET between 2011 and 2014&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
examined twice, the first time in the years 2014-2015 and for the second time in the years 2018-2019&#xD;
blood for laboratory tests&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
          </group>
          <group group_id="O2">
            <title>Control Group</title>
            <description>the control group is selected among healthy employees of the University Medical Centre Ljubljana and their relatives&#xD;
they are matched with the patient group for age and sex distribution and classical risk factors for cardiovascular disease&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
blood for laboratory tests&#xD;
examined twice, the first time in the years 2014-2015 and for the second time the in years 2018-2019&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
          </group>
        </group_list>
        <measure>
          <title>Carotid Artery Stiffness (Expressed by the Pulse Wave Velocity) in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period (From Baseline to Year 4).</title>
          <description>4-year change in carotid artery pulse wave velocity (PWV) measured in JAK2 V617F positive ET patients in comparison to healthy control subjects. All measurements were performed by echo-tracking ultrasound of the common carotid artery.&#xD;
Measurements were done at the common carotid artery 2 cm proximal to the bulb on both sides. PWV was calculated from the beta stiffness index (β, for description see above), as follows:&#xD;
PWV = √ ((β x P_min) / 2ρ)); ρ = 1050 kg/m³. PWV increases with arterial stiffness, a larger value means a worse outcome. PWV in adults ranges from 3 m/s to 12 m/s; values are strongly age dependent.</description>
          <units>m/s</units>
          <param>Mean</param>
          <dispersion>Standard Deviation</dispersion>
          <analyzed_list>
            <analyzed>
              <units>Participants</units>
              <scope>Measure</scope>
              <count_list>
                <count group_id="O1" value="36"/>
                <count group_id="O2" value="38"/>
              </count_list>
            </analyzed>
          </analyzed_list>
          <class_list>
            <class>
              <category_list>
                <category>
                  <measurement_list>
                    <measurement group_id="O1" value="0.72" spread="0.92"/>
                    <measurement group_id="O2" value="0.08" spread="0.72"/>
                  </measurement_list>
                </category>
              </category_list>
            </class>
          </class_list>
        </measure>
      </outcome>
      <outcome>
        <type>Primary</type>
        <title>Change of Carotid Artery Plaque Score in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period.</title>
        <description>Change in carotid plaque score (i.e., the number of carotid artery segments with detected plaques, reported on a scale from 0-6) in a 4 year observation period.&#xD;
A carotid plaque score 0 means that the common carotid, internal carotid and exteranal carotid artery on both sides are free from plaques. A score of 6 means that the common carotid, internal carotid and external carotid artery on both sides have plaques detected by ultrasound.</description>
        <time_frame>From Baseline (in 2014-2015) to Year 4 (in 2018-2019)</time_frame>
        <group_list>
          <group group_id="O1">
            <title>Patients With Essential Thrombocythemia</title>
            <description>essential thrombocythemia with JAK2 V617F positive mutation&#xD;
patients from the Department of Haematology at University Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET between 2011 and 2014&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
examined twice, the first time in the years 2014-2015 and for the second time in the years 2018-2019&#xD;
blood for laboratory tests&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
          </group>
          <group group_id="O2">
            <title>Control Group</title>
            <description>the control group is selected among healthy employees of the University Medical Centre Ljubljana and their relatives&#xD;
they are matched with the patient group for age and sex distribution and classical risk factors for cardiovascular disease&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
blood for laboratory tests&#xD;
examined twice, the first time in the years 2014-2015 and for the second time the in years 2018-2019&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
          </group>
        </group_list>
        <measure>
          <title>Change of Carotid Artery Plaque Score in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period.</title>
          <description>Change in carotid plaque score (i.e., the number of carotid artery segments with detected plaques, reported on a scale from 0-6) in a 4 year observation period.&#xD;
A carotid plaque score 0 means that the common carotid, internal carotid and exteranal carotid artery on both sides are free from plaques. A score of 6 means that the common carotid, internal carotid and external carotid artery on both sides have plaques detected by ultrasound.</description>
          <units>score on a scale</units>
          <param>Median</param>
          <dispersion>Inter-Quartile Range</dispersion>
          <analyzed_list>
            <analyzed>
              <units>Participants</units>
              <scope>Measure</scope>
              <count_list>
                <count group_id="O1" value="36"/>
                <count group_id="O2" value="38"/>
              </count_list>
            </analyzed>
          </analyzed_list>
          <class_list>
            <class>
              <category_list>
                <category>
                  <measurement_list>
                    <measurement group_id="O1" value="0" lower_limit="0" upper_limit="0"/>
                    <measurement group_id="O2" value="0" lower_limit="0" upper_limit="0"/>
                  </measurement_list>
                </category>
              </category_list>
            </class>
          </class_list>
        </measure>
      </outcome>
      <outcome>
        <type>Primary</type>
        <title>Change of Coronary Calcium Burden in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period.</title>
        <description>Change in the Agatston coronary artery calcium score in a 4 year observation period.&#xD;
Agatston score denotes the number of coronary artery calcifications weighted for the intensity of calcification.&#xD;
Grading of coronary artery disease (CAD) based od Agatston score goes as follows: no CAD = score 0, minimal CAD = score 1-10, mild CAD = score 11-100, moderate CAD = score 101-400, svere CAD = score &gt; 400.</description>
        <time_frame>From Baseline (in 2014-2015) to Year 4 (in 2018-2019)</time_frame>
        <group_list>
          <group group_id="O1">
            <title>Patients With Essential Thrombocythemia</title>
            <description>essential thrombocythemia with JAK2 V617F positive mutation&#xD;
patients from the Department of Haematology at University Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET between 2011 and 2014&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
examined twice, the first time in the years 2014-2015 and for the second time in the years 2018-2019&#xD;
blood for laboratory tests&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
          </group>
          <group group_id="O2">
            <title>Control Group</title>
            <description>the control group is selected among healthy employees of the University Medical Centre Ljubljana and their relatives&#xD;
they are matched with the patient group for age and sex distribution and classical risk factors for cardiovascular disease&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
blood for laboratory tests&#xD;
examined twice, the first time in the years 2014-2015 and for the second time the in years 2018-2019&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
          </group>
        </group_list>
        <measure>
          <title>Change of Coronary Calcium Burden in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period.</title>
          <description>Change in the Agatston coronary artery calcium score in a 4 year observation period.&#xD;
Agatston score denotes the number of coronary artery calcifications weighted for the intensity of calcification.&#xD;
Grading of coronary artery disease (CAD) based od Agatston score goes as follows: no CAD = score 0, minimal CAD = score 1-10, mild CAD = score 11-100, moderate CAD = score 101-400, svere CAD = score &gt; 400.</description>
          <units>units on a scale</units>
          <param>Median</param>
          <dispersion>Inter-Quartile Range</dispersion>
          <analyzed_list>
            <analyzed>
              <units>Participants</units>
              <scope>Measure</scope>
              <count_list>
                <count group_id="O1" value="36"/>
                <count group_id="O2" value="38"/>
              </count_list>
            </analyzed>
          </analyzed_list>
          <class_list>
            <class>
              <category_list>
                <category>
                  <measurement_list>
                    <measurement group_id="O1" value="0" lower_limit="0" upper_limit="15.85"/>
                    <measurement group_id="O2" value="1.6" lower_limit="0" upper_limit="14.10"/>
                  </measurement_list>
                </category>
              </category_list>
            </class>
          </class_list>
        </measure>
      </outcome>
      <outcome>
        <type>Primary</type>
        <title>Change of Digital Endothelial Function, Expressed as Reactive Hyperemia Index and Augmentation Index (AI), in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period.</title>
        <description>4-year change in Reactive Hyperemia Index (RHI) and 4-year change Augmentation Index (AI).&#xD;
RHI was calculated by the formula:&#xD;
RHI = (A/B) / (C/D) where A is the post-occlusion pulse wave amplitude (PWA) of the occluded hand, B the baseline PWA of the occluded hand, C the post-occlusion PWA of the contralateral hand, and D the baseline PWA of the contralateral hand. A higher RHI means better endothelial function (= good outcome). RHI in patients ranges from1-3. Arbitrarily, normal RHI is defined as =&gt; 1.67, and abnormal RHI as RHI =&lt; 1.66.&#xD;
The AI was determined from the shape of the arterial pulse wave by the EndoPAT 2000 software which distinguished between the primary pulse wave (P1) and the reflected pulse wave (P2) by the formula: AI = ((P2-P1)/P1) x 100. The results were normalized to a heart rate of 75/min. A higher AI means greater arterial stiffness (= bad outcome). AI &gt;0.4 is abnormal.</description>
        <time_frame>From Baseline (in 2014-2015) to Year 4 (in 2018-2019)</time_frame>
        <group_list>
          <group group_id="O1">
            <title>Patients With Essential Thrombocythemia</title>
            <description>essential thrombocythemia with JAK2 V617F positive mutation&#xD;
patients from the Department of Haematology at University Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET between 2011 and 2014&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
examined twice, the first time in the years 2014-2015 and for the second time in the years 2018-2019&#xD;
blood for laboratory tests&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
          </group>
          <group group_id="O2">
            <title>Control Group</title>
            <description>the control group is selected among healthy employees of the University Medical Centre Ljubljana and their relatives&#xD;
they are matched with the patient group for age and sex distribution and classical risk factors for cardiovascular disease&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
blood for laboratory tests&#xD;
examined twice, the first time in the years 2014-2015 and for the second time the in years 2018-2019&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
          </group>
        </group_list>
        <measure>
          <title>Change of Digital Endothelial Function, Expressed as Reactive Hyperemia Index and Augmentation Index (AI), in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period.</title>
          <description>4-year change in Reactive Hyperemia Index (RHI) and 4-year change Augmentation Index (AI).&#xD;
RHI was calculated by the formula:&#xD;
RHI = (A/B) / (C/D) where A is the post-occlusion pulse wave amplitude (PWA) of the occluded hand, B the baseline PWA of the occluded hand, C the post-occlusion PWA of the contralateral hand, and D the baseline PWA of the contralateral hand. A higher RHI means better endothelial function (= good outcome). RHI in patients ranges from1-3. Arbitrarily, normal RHI is defined as =&gt; 1.67, and abnormal RHI as RHI =&lt; 1.66.&#xD;
The AI was determined from the shape of the arterial pulse wave by the EndoPAT 2000 software which distinguished between the primary pulse wave (P1) and the reflected pulse wave (P2) by the formula: AI = ((P2-P1)/P1) x 100. The results were normalized to a heart rate of 75/min. A higher AI means greater arterial stiffness (= bad outcome). AI &gt;0.4 is abnormal.</description>
          <units>units on a scale</units>
          <param>Mean</param>
          <dispersion>Standard Deviation</dispersion>
          <analyzed_list>
            <analyzed>
              <units>Participants</units>
              <scope>Measure</scope>
              <count_list>
                <count group_id="O1" value="36"/>
                <count group_id="O2" value="38"/>
              </count_list>
            </analyzed>
          </analyzed_list>
          <class_list>
            <class>
              <title>Reactive Hyperemia Index (RHI)</title>
              <category_list>
                <category>
                  <measurement_list>
                    <measurement group_id="O1" value="-0.02" spread="0.77"/>
                    <measurement group_id="O2" value="-0.26" spread="0.89"/>
                  </measurement_list>
                </category>
              </category_list>
            </class>
            <class>
              <title>Augmentation Index (AI)</title>
              <category_list>
                <category>
                  <measurement_list>
                    <measurement group_id="O1" value="3.56" spread="16.88"/>
                    <measurement group_id="O2" value="0.46" spread="20.19"/>
                  </measurement_list>
                </category>
              </category_list>
            </class>
          </class_list>
        </measure>
      </outcome>
      <outcome>
        <type>Primary</type>
        <title>The JAK2 V617F Mutation Burden in Patients With JAK2 V617F Positive ET.</title>
        <description>Change in JAK2 V617F mutation burden in the 4-year observation period. Ipsogen JAK2 MutaQuant Kit, Qiagen (USA) was used for the detection of JAK2 V617F mutation and quantification of its burden in genomic DNA, which was extracted from granulocytes in peripheral blood. Real-time quantitative polymerase chain reaction (qPCR) was used with double-dye oligonucleotide hydrolysis principle. The JAK2 V617F burden was defined as the percentage of JAK2 V617F mutated alleles in total genomic DNA.</description>
        <time_frame>At baseline (in 2014- 2015) and at the second visit (in 2018-2019)</time_frame>
        <population>The control group was not included in the comparison because they lack JAK2 V617F mutation as they are healthy subjects with no mutation. We examine the association between mutation burden (where the mutation is necessary) and coronary calcium burden.</population>
        <group_list>
          <group group_id="O1">
            <title>Patients With Essential Thrombocythemia</title>
            <description>essential thrombocythemia with JAK2 V617F positive mutation&#xD;
patients from the Department of Haematology at University Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET between 2011 and 2014&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
examined twice, the first time in the years 2014-2015 and for the second time in the years 2018-2019&#xD;
blood for laboratory tests&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning</description>
          </group>
        </group_list>
        <measure>
          <title>The JAK2 V617F Mutation Burden in Patients With JAK2 V617F Positive ET.</title>
          <description>Change in JAK2 V617F mutation burden in the 4-year observation period. Ipsogen JAK2 MutaQuant Kit, Qiagen (USA) was used for the detection of JAK2 V617F mutation and quantification of its burden in genomic DNA, which was extracted from granulocytes in peripheral blood. Real-time quantitative polymerase chain reaction (qPCR) was used with double-dye oligonucleotide hydrolysis principle. The JAK2 V617F burden was defined as the percentage of JAK2 V617F mutated alleles in total genomic DNA.</description>
          <population>The control group was not included in the comparison because they lack JAK2 V617F mutation as they are healthy subjects with no mutation. We examine the association between mutation burden (where the mutation is necessary) and coronary calcium burden.</population>
          <units>percentage of mutant allele</units>
          <param>Median</param>
          <dispersion>Inter-Quartile Range</dispersion>
          <analyzed_list>
            <analyzed>
              <units>Participants</units>
              <scope>Measure</scope>
              <count_list>
                <count group_id="O1" value="36"/>
              </count_list>
            </analyzed>
          </analyzed_list>
          <class_list>
            <class>
              <title>JAK2 V617F mutation burden at baseline</title>
              <category_list>
                <category>
                  <measurement_list>
                    <measurement group_id="O1" value="29.95" lower_limit="7.32" upper_limit="43.33"/>
                  </measurement_list>
                </category>
              </category_list>
            </class>
            <class>
              <title>JAK2 V617F mutation burden at the second visit</title>
              <category_list>
                <category>
                  <measurement_list>
                    <measurement group_id="O1" value="11.60" lower_limit="0" upper_limit="26.63"/>
                  </measurement_list>
                </category>
              </category_list>
            </class>
          </class_list>
        </measure>
      </outcome>
    </outcome_list>

    <reported_events>
      <time_frame>From Baseline (in 2014- 2015) to Year 4 (2018-2019)</time_frame>
      <desc>There were no adverse events.</desc>
      <group_list>
        <group group_id="E1">
          <title>Patients With Essential Thrombocythemia</title>
          <description>essential thrombocythemia with JAK2 V617F positive mutation&#xD;
patients from the Department of Haematology at University Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET between 2011 and 2014&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
examined twice, the first time in the years 2014-2015 and for the second time in the years 2018-2019&#xD;
blood for laboratory tests&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
        </group>
        <group group_id="E2">
          <title>Control Group</title>
          <description>the control group is selected among healthy employees of the University Medical Centre Ljubljana and their relatives&#xD;
they are matched with the patient group for age and sex distribution and classical risk factors for cardiovascular disease&#xD;
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)&#xD;
all signed the informed consent&#xD;
blood for laboratory tests&#xD;
examined twice, the first time in the years 2014-2015 and for the second time the in years 2018-2019&#xD;
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning&#xD;
imaging</description>
        </group>
      </group_list>
      <serious_events>
        <category_list>
          <category>
            <title>Total</title>
            <event_list>
              <event>
                <sub_title>Total, all-cause mortality</sub_title>
                <counts group_id="E1" subjects_affected="0" subjects_at_risk="36"/>
                <counts group_id="E2" subjects_affected="0" subjects_at_risk="38"/>
              </event>
              <event>
                <sub_title>Total, serious adverse events</sub_title>
                <counts group_id="E1" subjects_affected="0" subjects_at_risk="36"/>
                <counts group_id="E2" subjects_affected="0" subjects_at_risk="38"/>
              </event>
            </event_list>
          </category>
        </category_list>
      </serious_events>
      <other_events>
        <frequency_threshold>0</frequency_threshold>
        <category_list>
          <category>
            <title>Total</title>
            <event_list>
              <event>
                <sub_title>Total, other adverse events</sub_title>
                <counts group_id="E1" subjects_affected="0" subjects_at_risk="36"/>
                <counts group_id="E2" subjects_affected="0" subjects_at_risk="38"/>
              </event>
            </event_list>
          </category>
        </category_list>
      </other_events>
    </reported_events>

    <certain_agreements>
      <pi_employee>Principal Investigators are NOT employed by the organization sponsoring the study.</pi_employee>
      <restrictive_agreement>There is NOT an agreement between Principal Investigators and the Sponsor (or its agents) that restricts the PI's rights to discuss or publish trial results after the trial is completed. </restrictive_agreement>
    </certain_agreements>
    <limitations_and_caveats>the relatively small number of participants (patients were recruited from a single tertiary medical centre)&#xD;
the subjects were not examined at precisely the same time interval between both visit</limitations_and_caveats>
    <point_of_contact>
      <name_or_title>dr. Aleš Blinc, MD, PhD</name_or_title>
      <organization>University Medical Centre Ljubljana</organization>
      <phone>+38615227124</phone>
      <email>ales.blinc@kclj.si</email>
    </point_of_contact>
  </clinical_results>
</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/NCT03828422
org study id: TP20180038
nct id: NCT03828422
lead sponsor: 
has dmc: No
is fda regulated drug: No
is fda regulated device: No
The aim of the study is to examine (a) whether patients with JAK2 V617F positive ET in
      comparison to age-and sex-matched, apparently healthy control subjects show more advanced
      progression of arterial stiffness, pulse-wave velocity and coronary calcium score in a 4 year
      observation period, and (b) whether the burden of JAK2 V617F mutation correlates with the
      measured vascular parameters. All subjects will be examined twice. The first visit already
      took place between the years 2014 - 2015 and the second visit will take place between
      2018-2019. All participants will have signed their informed consent before entering the
      study. Each visit will consist of completing a structured questionnaire (on personal and
      family medical history, risk factors for CVD and medication), physical examination, donating
      a blood sample for laboratory tests and undergoing carotid ultrasound and coronary calcium
      measurement oft the extent of coronary artery calcification. At the first and the second
      examination the JAK2 V617F allele burden, i.e. the percentage of mutated alleles, will be
      determined from genomic DNA in peripheral blood.
1. Patients and control subjects

           Patients are selected from the database of the Department of Haematology at University
           Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET
           between 2011 and 2014. There were 124 JAK2 positive ET patients, among those 61 did not
           have a personal history of clinically manifest atherosclerotic vascular disease
           (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease,
           transient ischemic attack or ischemic stroke). 40 patients (14 male and 26 female) with
           JAK2 V617F positive ET without clinically apparent cardiovascular disease signed the
           informed consent and were enrolled in the study in 2014 - 2015 for the first examination
           and 36 (12 male and 24 female) of them are expected to participate also in 2018-19.

           The control group is selected among healthy employees of the University Medical Centre
           Ljubljana and their relatives. It is matched with the patient group for age and sex
           distribution and classical risk factors for cardiovascular disease. 42 individuals (16
           male and 26 female) participated in the first examination and at least 38 (14 male and
           24 female) subjects are expected to paticipate in the second examination.

        2. Baseline measurements

           Each participant will complete a questionnaire about family history of cardiovascular
           diseases, personal medical history, smoking status and medications. A physical
           examination will be performed, including measurements of height, weight, waist
           circumference, systolic and diastolic blood pressure. Blood will be sampled for
           laboratory analysis of complete blood cell count, electrolytes, serum lipids, liver
           function tests, urea, creatinine, and inflammatory markers.

           The 10-year risk of coronary heart disease (CHD) and general cardiovascular disease
           (CVD) will be calculated using the Framingham risk equation.

        3. JAK2 V617F/G1849T allele burden

           The ipsogen JAK2 MutaQuant Kit, Qiagen (ZDA) (Ref: No. 673523) will be used for the
           detection and quantification of JAK2 V617F/G1849T allele in genomic DNA extracted from
           peripheral blood of patients and also control subjects. A SNP specific primer
           selectively amplifies the JAK2 V617F allele which is detected with a real-time qPCR
           instrument that quantifies the PCR products. The JAK2 V617F allele burden will be
           calculated and expressed as the percentage of JAK2 V617F mutated alleles throughout the
           whole JAK2 record. Analysis will be done done in laboratory of the Department of
           Haematology at University Medical Centre Ljubljana for all samples.

        4. Carotid Artery Ultrasound Examination

           For examination of the extracranial carotid arteries an ultrasound machine Aloka
           Prosound α7 (Hitachi Aloka Medical, Ltd., Japan) will be used with a linear vascular
           probe working at a frequency of 5-13 MHz. The common, internal and external carotid
           arteries on both sided will be examined. At each examination, the measurements will be
           done twice and their average values will be calculated. The ultrasound examination will
           be done by the same ultrasonographer at both visits of all participants.

           The intima-media thickness will be measured 2 cm proximal to the bulb of common carotid
           artery on both sides. Screening of the extracranial carotid arteries for atherosclerotic
           plaques will be performed, with a plaque definition of a focal lesion, exceeding the
           intima-media thickness for at least 50% or reaching an absolute thickness of at least
           1.5 mm in two orthogonal projections. Scoring of atherosclerotic plaques is done
           according to the methodology from the Rotterdam Study. The presence of at least one
           plaque in each segment of the extracranial carotid arterial bed, (divided into the
           common carotid artery and the bulb, the internal carotid artery and the external carotid
           artery) on either sides is scored 1 point. Thus, the carotid plaque score ranges from 0
           (absence of plaques) to 6 (plaques present in all segments on both sides).

           Echo-tracking of the common carotid artery will be used to assess arterial stiffness
           expressed by the β-stiffness index and the pulse wave velocity. Measurements will be
           done at the common carotid artery 2 cm proximal to the bulb on both sides. The
           β-stiffness index will be calculated as: β = ln (P_max / P_min) / [(D_max - D_min /
           D_min)], where P_max = the systolic blood pressure, P_min = the diastolic pressure;
           D_max = the maximum arterial diameter and D_min = the minimal arterial diameter. Pulse
           wave velocity (PWV) will be calculated as: PWV = √ ((β x P_min) / 2ρ)); ρ = 1050 kg/m³.

        5. Assessment of Endothelial Function of the Digital Arteries

           Endothelial function of the digital arteries will be measured by digital plethymography
           with EndoPat 2000, Itamar Medical REF, Caesarea, Israel, Software Version 3.3.x and
           expressed as the Reactive Hyperaemia Index (RHI) and the Augmentation Index (AI).
           Changes in arterial tone are elicited by creating a downstream hyperemic response
           induced by a standard 5-minute occlusion of the brachial artery (using a blood pressure
           cuff, inflated to 60 mmHg above the arterial blood pressure). When the cuff is released,
           the surge of blood flow causes endothelium-dependent flow mediated dilatation (FMD)
           which is manifested as reactive hyperemia. A post-occlusion to pre-occlusion ratio is
           calculated by EndoPAT software and the and A) are determined. The results are normalized
           to a heart rate of 75/min.

        6. Measurement of the Coronary Artery Calcium Burden

           The calcium burden of coronary arteries is measured with a Biograph M 128-row PET-CT
           scanner (Siemens, Erlangen, Germany). Scanning is done from the base to the apex of the
           heart. A non-contrast protocol with sequential prospective ECG triggering is used. The
           rotation time is 0.33 sec, with a tube voltage of 120 kV, CARE Dose 4D and slice
           thickness 3 mm, with no slice overlap. Post-processing is done on the Syngo Leonardo
           workstation. The coronary calcium burden is expressed as the Agatston score. Evaluation
           of the dataset of every study subject is done three times and the average value is used
           for further analysis.

        7. Statistical analysis

      All sets of data will be tested for normality of distribution using the normal-quintile plot,
      calculating the correlation coefficient and checking it for the critical value that would
      warrant rejection of normal distribution with an α-error probability of 0.05.

      Normally distributed data will be presented as mean and standard deviation, while
      non-normally distributed data will be presented as median and range between the 1st and 3rd
      quartile.

      Differences between subjects with ET and control subjects at the first and second examination
      will be tested by the chi-square test for discrete variables, for normally distributed
      continuous variables by the paired Student's t-test for independent samples, and for
      non-normally distributed continuous variables by the Mann-Whitney test for independent
      samples. P-values of < 0.05 will be considered significant with a correction for multiple
      comparisons.

      Changes in vascular function and morphology in the 4-year observation period will be compared
      between the two groups by the log rank test. P-values of < 0.05 will be considered
      significant.

      The association between the parameters of vascular function / morphology and the JAK2 V617F
      allele burden will be assessed by the Pearson correlation coefficient.
observational model: Case-Control
time perspective: Prospective
measure: Carotid Artery Stiffness (Expressed by Beta-stiffness Index ) in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period (From Baseline to Year 4).
time frame: From Baseline (in 2014-2015) to Year 4 (in 2018-2019)
description: 4-year change carotid artery in Beta-stiffness Index measured in JAK2 V617F positive ET patients in comparison to healthy control subjects. All measurements were performed by echo-tracking ultrasound of the common carotid artery.
Echo-tracking of the common carotid artery wall was used to assess arterial stiffness expressed by the β-stiffness index. Measurement swere done at the common carotid artery 2 cm proximal to the bulb on both sides. The β-stiffness index was calculated as: β = ln (P_max / P_min) / [(D_max - D_min / D_min)] (P_max = the systolic blood pressure, P_min = the diastolic pressure; D_max = the maximum arterial diameter and D_min = the minimal arterial diameter). In the adult population, the values of β range from 4 - 10 and increase with age. A larger value of β means increased arterial stiffness, i.e., a worse outcome.
measure: Carotid Artery Stiffness (Expressed by the Pulse Wave Velocity) in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period (From Baseline to Year 4).
time frame: From Baseline (in 2014-2015) to Year 4 (in 2018-2019)
description: 4-year change in carotid artery pulse wave velocity (PWV) measured in JAK2 V617F positive ET patients in comparison to healthy control subjects. All measurements were performed by echo-tracking ultrasound of the common carotid artery.
Measurements were done at the common carotid artery 2 cm proximal to the bulb on both sides. PWV was calculated from the beta stiffness index (β, for description see above), as follows:
PWV = √ ((β x P_min) / 2ρ)); ρ = 1050 kg/m³. PWV increases with arterial stiffness, a larger value means a worse outcome. PWV in adults ranges from 3 m/s to 12 m/s; values are strongly age dependent.
measure: Change of Carotid Artery Plaque Score in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period.
time frame: From Baseline (in 2014-2015) to Year 4 (in 2018-2019)
description: Change in carotid plaque score (i.e., the number of carotid artery segments with detected plaques, reported on a scale from 0-6) in a 4 year observation period.
A carotid plaque score 0 means that the common carotid, internal carotid and exteranal carotid artery on both sides are free from plaques. A score of 6 means that the common carotid, internal carotid and external carotid artery on both sides have plaques detected by ultrasound.
measure: Change of Coronary Calcium Burden in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period.
time frame: From Baseline (in 2014-2015) to Year 4 (in 2018-2019)
description: Change in the Agatston coronary artery calcium score in a 4 year observation period.
Agatston score denotes the number of coronary artery calcifications weighted for the intensity of calcification.
Grading of coronary artery disease (CAD) based od Agatston score goes as follows: no CAD = score 0, minimal CAD = score 1-10, mild CAD = score 11-100, moderate CAD = score 101-400, svere CAD = score > 400.
measure: Change of Digital Endothelial Function, Expressed as Reactive Hyperemia Index and Augmentation Index (AI), in JAK2 V617F Positive ET Patients in Comparison to Healthy Control Subjects in a 4-year Period.
time frame: From Baseline (in 2014-2015) to Year 4 (in 2018-2019)
description: 4-year change in Reactive Hyperemia Index (RHI) and 4-year change Augmentation Index (AI).
RHI was calculated by the formula:
RHI = (A/B) / (C/D) where A is the post-occlusion pulse wave amplitude (PWA) of the occluded hand, B the baseline PWA of the occluded hand, C the post-occlusion PWA of the contralateral hand, and D the baseline PWA of the contralateral hand. A higher RHI means better endothelial function (= good outcome). RHI in patients ranges from1-3. Arbitrarily, normal RHI is defined as => 1.67, and abnormal RHI as RHI =< 1.66.
The AI was determined from the shape of the arterial pulse wave by the EndoPAT 2000 software which distinguished between the primary pulse wave (P1) and the reflected pulse wave (P2) by the formula: AI = ((P2-P1)/P1) x 100. The results were normalized to a heart rate of 75/min. A higher AI means greater arterial stiffness (= bad outcome). AI >0.4 is abnormal.
measure: The JAK2 V617F Mutation Burden in Patients With JAK2 V617F Positive ET.
time frame: At baseline (in 2014- 2015) and at the second visit (in 2018-2019)
description: Change in JAK2 V617F mutation burden in the 4-year observation period. Ipsogen JAK2 MutaQuant Kit, Qiagen (USA) was used for the detection of JAK2 V617F mutation and quantification of its burden in genomic DNA, which was extracted from granulocytes in peripheral blood. Real-time quantitative polymerase chain reaction (qPCR) was used with double-dye oligonucleotide hydrolysis principle. The JAK2 V617F burden was defined as the percentage of JAK2 V617F mutated alleles in total genomic DNA.
arm group label: patients with essential thrombocythemia
description: essential thrombocythemia with JAK2 V617F positive mutation
patients from the Department of Haematology at University Medical Centre Ljubljana, Slovenia, who were diagnosed with JAK2 V617F positive ET between 2011 and 2014
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)
all signed the informed consent
examined twice, the first time in the years 2014-2015 and for the second time in the years 2018-2019
blood for laboratory tests
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning
arm group label: control group
description: the control group is selected among healthy employees of the University Medical Centre Ljubljana and their relatives
they are matched with the patient group for age and sex distribution and classical risk factors for cardiovascular disease
no personal history of clinically manifest atherosclerotic vascular disease (myocardial infarction, angina pectoris, peripheral arterial disease, aortic disease, transient ischemic attack or ischemic stroke)
all signed the informed consent
blood for laboratory tests
examined twice, the first time in the years 2014-2015 and for the second time the in years 2018-2019
imaging and functional examination: ultrasound examination, EndoPat plethysmography, coronary artery calcium scanning
intervention type: Diagnostic Test
intervention name: imaging
arm group label: control group
arm group label: patients with essential thrombocythemia
other name: EndoPat plethysmography
other name: coronary artery calcium scanning
other name: blood laboratory tests
other name: ultrasound examination
The ipsogen JAK2 MutaQuant Kit, Qiagen (ZDA) (Ref: No. 673523) will be used for the detection
      and quantification of JAK2 V617F/G1849T allele in genomic DNA extracted from peripheral blood
      of patients and also control subjects. A SNP specific primer that selectively amplifies JAK2
      V617F allele will be used with a real-time qPCR instrument which accurately quantifies the
      PCR products.
study pop: 
sampling method: Probability Sample
criteria: 
gender: All
minimum age: 18 Years
maximum age: N/A
healthy volunteers: Accepts Healthy Volunteers
last name: Ales Blinc, MD
role: Study Director
affiliation: University Medical Centre Ljubljana
citation: Vrtovec M, Anzic A, Zupan IP, Zaletel K, Blinc A. Carotid Artery Stiffness, Digital Endothelial Function, and Coronary Calcium in Patients with Essential Thrombocytosis, Free of Overt Atherosclerotic Disease. Radiol Oncol. 2017 Jan 14;51(2):203-210. doi: 10.1515/raon-2017-0006. eCollection 2017 Jun.
PMID: 28740456
citation: Anzic Drofenik A, Vrtovec M, Bozic Mijovski M, Sever M, Preloznik Zupan I, Kejzar N, Blinc A. Progression of coronary calcium burden and carotid stiffness in patients with essential thrombocythemia associated with JAK2 V617F mutation. Atherosclerosis. 2020 Mar;296:25-31. doi: 10.1016/j.atherosclerosis.2020.01.001. Epub 2020 Jan 11.
PMID: 32005002
responsible party type: Principal Investigator
investigator affiliation: University Medical Centre Ljubljana
investigator full name: Ales Blinc
investigator title: Ales Blinc, MD, PhD, Head, Dept of Vascular Diseases
mesh term: Atherosclerosis
mesh term: Thrombocytosis
mesh term: Thrombocythemia, Essential
mesh term: Calcium
sharing ipd: Undecided
provided document: 
participant flow: 
baseline: 
outcome list: 
reported events: 
certain agreements: 
limitations and caveats: the relatively small number of participants (patients were recruited from a single tertiary medical centre)
the subjects were not examined at precisely the same time interval between both visit
point of contact: 