<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/NCT04496154</url>
  </required_header>
  <id_info>
    <org_study_id>2013-14 and 2014-756</org_study_id>
    <secondary_id>NUT273741</secondary_id>
    <nct_id>NCT04496154</nct_id>
  </id_info>
  <brief_title>Omega-3 to Reduce Diabetes Risk in Subjects With High Number of Particles That Carry &quot;Bad Cholesterol&quot; in the Blood</brief_title>
  <official_title>The Inflammasome and Dysfunctional Adipose Tissue: Why Should apoB-lipoproteins be Targeted in Humans</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>May Faraj, PDt, PhD</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>Canadian Institutes of Health Research (CIHR)</agency>
      <agency_class>Other</agency_class>
    </collaborator>
  </sponsors>
  <source>Institut de Recherches Cliniques de Montreal</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>
      In this project, investigators explored the role of the particles that carry &quot;bad&#xD;
      cholesterol&quot; in the blood (termed LDL) that are known to promote heart disease, in the&#xD;
      promotion of type 2 diabetes (T2D) in humans. In specific, they investigated how these&#xD;
      particles may induce the activation of an immune pathway in human fat tissue leading to&#xD;
      multiple anomalies that favors T2D. They also explored whether omega-3 fatty acids, which are&#xD;
      the type of fat found in fish oils can counterbalance the negative effects of LDL in fat&#xD;
      tissue, thus providing a natural way to help reduce the risk for T2D in subjects with&#xD;
      elevated blood LDL.&#xD;
&#xD;
      To do so, 41 subjects who were free of disease or medication affecting metabolism were&#xD;
      enrolled at the Montreal Clinical Research Institute between 2013 and 2019 and were placed on&#xD;
      an intervention with omega-3 fatty acids supplementation for 12 weeks (2.7 g/day, Triple&#xD;
      Strength Omega-3 from Webbers Naturals). Investigators examined the effects of LDL and&#xD;
      omega-3 on risk factors for T2D before and after the intervention in the whole body and&#xD;
      specifically in fat tissue biopsies taken from the hip region. Eighty percent of the subjects&#xD;
      who were enrolled into the study completed the intervention.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      Diabetes-attributed deaths, mostly type 2 diabetes (T2D), total more than 40,000 per year,&#xD;
      out of which 80% are secondary to cardiovascular disease and stroke. Research from the&#xD;
      investigators' lab and others suggest that elevated atherogenic apoB-lipoproteins may not be&#xD;
      a mere consequence of T2D but also a cause. They reported that high number of&#xD;
      apoB-lipoproteins (apoB) may directly induce subcutaneous white adipose tissue dysfunction&#xD;
      and related risk factors for T2D in humans; however underlying mechanisms were yet unclear.&#xD;
&#xD;
      Strong evidence implicates a specific innate immunity complex, the NLRP3 inflammasome (NLRP3&#xD;
      for Nucleotide-binding domain and Leucine-rich repeat Receptor, containing a Pyrin domain 3)&#xD;
      in white adipose tissue dysfunction and associated metabolic anomalies in mice and humans.&#xD;
      Preliminary evidence from the investigators lab and their collaborator (Dr Maya Saleh, at&#xD;
      McGill University) indicated that native apoB-lipoproteins activate the NLRP3 inflammasome in&#xD;
      murine bone marrow derived macrophages. On the other hand, omega-3 fatty acids were reported&#xD;
      to inhibit the NLRP3 inflammasome in immune cells.&#xD;
&#xD;
      Thus the hypotheses that were examined in this study were that:&#xD;
&#xD;
      Primary hypothesis:&#xD;
&#xD;
        1. Subjects with high plasma apoB have higher NLRP3 inflammasome activity in white adipose&#xD;
           tissue indicated by higher interleukin 1 beta (IL-1β) secretion.&#xD;
&#xD;
           Higher white adipose tissue IL-1β secretion was expected to be associated with risk&#xD;
           factors for T2D documented in subjects with high plasma apoB, namely white adipose&#xD;
           tissue dysfunction, systemic inflammation, postprandial hypertriglyceridemia, insulin&#xD;
           resistance and hyperinsulinemia&#xD;
&#xD;
           Secondary hypotheses:&#xD;
&#xD;
        2. Twelve-week supplementation with omega-3 fatty acids (2.7 g/day, eicosapentaenoic acid&#xD;
           (EPA) and docosahexaenoic (DHA), 2:1) induces a greater reduction in white adipose&#xD;
           tissue NLRP3 inflammasome activity and related risk factors for T2D in subjects with&#xD;
           high plasma apoB.&#xD;
&#xD;
        3. Subject native LDL (representing &gt; 90% of apoB-lipoproteins) directly prime and/or&#xD;
           activate the NLRP3 inflammasome in subject own white adipose tissue ex vivo, while&#xD;
           EPA/DHA inhibit it.&#xD;
&#xD;
      Forty-one subjects (34% men) were enrolled in the study, of whom 33 subjects completed the&#xD;
      12-week omega-3 intervention (drop out/exclusion rate = 20%). Baseline and post-intervention&#xD;
      risk factors for T2D measured in these subjects were; white adipose tissue and systemic NLRP3&#xD;
      inflammasome activity, dietary fat clearance (after a high fat meal), adipose tissue function&#xD;
      (ex vivo after a subcutaneous adipose tissue needle biopsy), and insulin secretion and&#xD;
      sensitivity (by gold-standard Botnia clamp technique).&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Completed</overall_status>
  <start_date type="Actual">September 5, 2013</start_date>
  <completion_date type="Actual">February 24, 2020</completion_date>
  <primary_completion_date type="Actual">January 22, 2020</primary_completion_date>
  <phase>N/A</phase>
  <study_type>Interventional</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <allocation>N/A</allocation>
    <intervention_model>Single Group Assignment</intervention_model>
    <primary_purpose>Prevention</primary_purpose>
    <masking>None (Open Label)</masking>
  </study_design_info>
  <primary_outcome>
    <measure>Fasting white adipose tissue NLRP3 inflammasome activation</measure>
    <time_frame>Baseline</time_frame>
    <description>White adipose tissue medium accumulation of interleukin 1 beta (IL-1β) ex vivo over 4 hours (pg/mg tissue by AlphaLISA)</description>
  </primary_outcome>
  <secondary_outcome>
    <measure>Fasting white adipose tissue NLRP3 inflammasome activation</measure>
    <time_frame>Change at 12 weeks from baseline</time_frame>
    <description>White adipose tissue medium accumulation of interleukin 1 beta (IL-1β) ex vivo over 4 hours (pg/mg tissue by AlphaLISA)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>White adipose tissue inflammation profile</measure>
    <time_frame>Baseline</time_frame>
    <description>Fasting and 4 hour-postprandial change in NLRP3 inflammasome related inflammatory parameters; including protein and gene expression of IL1B and NLRP3 (by immunohistochemistry and RT-PCR), secretion of IL-1β (per mg tissue by AlphaLISA) and macrophage infiltration of white adipose tissue (by RT-PCR)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>White adipose tissue inflammation profile</measure>
    <time_frame>Change at 12 weeks from baseline</time_frame>
    <description>Fasting and 4 hour-postprandial change in NLRP3 inflammasome related inflammatory parameters; including protein and gene expression of IL1B and NLRP3 (by immunohistochemistry and RT-PCR), secretion of IL-1β (per mg tissue by AlphaLISA) and macrophage infiltration of white adipose tissue (by RT-PCR)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>White adipose tissue function ex vivo</measure>
    <time_frame>Baseline</time_frame>
    <description>Fasting in situ lipoprotein lipase activity (nmol 3H-triglyceride/mg tissue)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>White adipose tissue function ex vivo</measure>
    <time_frame>Change at 12 weeks from baseline</time_frame>
    <description>Fasting in situ lipoprotein lipase activity (nmol 3H-triglyceride/mg tissue)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Postprandial fat metabolism</measure>
    <time_frame>Baseline</time_frame>
    <description>Area under the 6 hour time curve of plasma triglycerides (mmol/hour) after a high-fat meal (66% fat)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Postprandial fat metabolism</measure>
    <time_frame>Change at 12 weeks from baseline</time_frame>
    <description>Area under the 6 hour time curve of plasma triglycerides (mmol/hour) after a high-fat meal (66% fat)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Systemic inflammation</measure>
    <time_frame>Baseline</time_frame>
    <description>Fasting plasma inflammatory parameters including IL-1Ra and IL-1β (pg/mL by AlphaLISA)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Systemic inflammation</measure>
    <time_frame>Change at 12 weeks from baseline</time_frame>
    <description>Fasting plasma inflammatory parameters including IL-1Ra and IL-1β (pg/mL by AlphaLISA)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Insulin sensitivity and secretion</measure>
    <time_frame>Baseline</time_frame>
    <description>Glucose-induced insulin secretion (uU/mL/min) and insulin sensitivity (glucose infusion rate mg/kg/min) measured by Botnia clamp</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Insulin sensitivity and secretion</measure>
    <time_frame>Change at 12 weeks from baseline</time_frame>
    <description>Glucose-induced insulin secretion (uU/mL/min) and insulin sensitivity (glucose infusion rate mg/kg/min) measured by Botnia clamp</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Fatty acid profile in the phospholipid fraction of plasma and red blood cells</measure>
    <time_frame>Baseline</time_frame>
    <description>(As μmol/L by gas chromatography mass spectrometry)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Fatty acid profile in the phospholipid fraction of plasma and red blood cells</measure>
    <time_frame>Change at 12 weeks from baseline</time_frame>
    <description>(As μmol/L by gas chromatography mass spectrometry)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Body composition</measure>
    <time_frame>Baseline</time_frame>
    <description>Fat and lean body mass (as kg by dual energy x-ray absorptiometry)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Body composition</measure>
    <time_frame>Change at 12 weeks from baseline</time_frame>
    <description>Fat and lean body mass (as kg by dual energy x-ray absorptiometry)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Energy expenditure</measure>
    <time_frame>Baseline</time_frame>
    <description>Fasting and postprandial during the 6-hour high fat meal challenge (as kcal/hr using indirect calorimetry)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Energy expenditure</measure>
    <time_frame>Change at 12 weeks from baseline</time_frame>
    <description>Fasting and postprandial during the 6-hour high fat meal challenge (as kcal/hr using indirect calorimetry)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Energy intake</measure>
    <time_frame>Baseline</time_frame>
    <description>(Average of 3 day energy intake as kcal/day collected by 3-day dietary records)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Energy intake</measure>
    <time_frame>Change at 12 weeks from baseline</time_frame>
    <description>(Average of 3 day energy intake as kcal/day collected by 3-day dietary records)</description>
  </secondary_outcome>
  <other_outcome>
    <measure>Post-hoc analyses of white adipose tissue receptors for apoB-lipoproteins</measure>
    <time_frame>Baseline</time_frame>
    <description>Fasting and 4 hour-postprandial change in white adipose tissue surface-expression LDLR and CD36 (% of control by immunohistochemistry in white adipose tissue slides)</description>
  </other_outcome>
  <other_outcome>
    <measure>Post-hoc analyses of white adipose tissue receptors for apoB-lipoproteins</measure>
    <time_frame>Change at 12 weeks from baseline</time_frame>
    <description>Fasting and 4 hour-postprandial change in white adipose tissue surface-expression LDLR and CD36 (% of control by immunohistochemistry in white adipose tissue slides)</description>
  </other_outcome>
  <other_outcome>
    <measure>Post-hoc analyses of white adipose tissue genes related to lipid and lipoprotein metabolism</measure>
    <time_frame>Baseline</time_frame>
    <description>Fasting and 4-hour postprandial change in gene expression including LDLR, CD36, SREBP1, SREBP2 and HMGCR (by RT-PCR)</description>
  </other_outcome>
  <other_outcome>
    <measure>Post-hoc analyses of white adipose tissue genes related to lipid and lipoprotein metabolism</measure>
    <time_frame>Change at 12 weeks from baseline</time_frame>
    <description>Fasting and 4-hour postprandial change in gene expression including LDLR, CD36, SREBP1, SREBP2 and HMGCR (by RT-PCR)</description>
  </other_outcome>
  <number_of_arms>1</number_of_arms>
  <enrollment type="Actual">41</enrollment>
  <condition>Type 2 Diabetes</condition>
  <condition>Insulin Sensitivity/Resistance</condition>
  <condition>Inflammatory Response</condition>
  <condition>Fatty Acids, Omega-3</condition>
  <arm_group>
    <arm_group_label>Omega-3 fatty acids</arm_group_label>
    <arm_group_type>Experimental</arm_group_type>
    <description>3 oral softgels (600 mg EPA and 300 mg DHA / softgel), Triple Strength Omega-3 from Webber Naturals</description>
  </arm_group>
  <intervention>
    <intervention_type>Dietary Supplement</intervention_type>
    <intervention_name>Omega-3 fatty acids (2.7 g/d, EPA:DHA, 2:1)</intervention_name>
    <description>Triple Strength Omega-3 from Webber Naturals</description>
    <arm_group_label>Omega-3 fatty acids</arm_group_label>
  </intervention>
  <eligibility>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
        Men and post-menopausal women:&#xD;
&#xD;
          -  Having a body mass index (BMI) &gt; 20 kg/m2&#xD;
&#xD;
          -  Aged between 45 and 74 years&#xD;
&#xD;
          -  Having confirmed menopausal status (FSH ≥ 30 U/l)&#xD;
&#xD;
          -  Non-smoker&#xD;
&#xD;
          -  Sedentary (less than 2 hours of structured physical exercise (ex: sports club) per&#xD;
             week)&#xD;
&#xD;
          -  Low alcohol consumption: less than 2 alcoholic drinks/day&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  Subjects with elevated risk of cardiovascular disease (≥ 20% of calculated Framingham&#xD;
             Risk Score) who require immediate medical intervention by lipid-lowering agents OR who&#xD;
             cannot be placed on a 4 weeks wash-out period from their lipid-lowering medication at&#xD;
             screening (i.e. upon admission to IRCM clinic).&#xD;
&#xD;
          -  Subjects with systolic blood pressure &gt; 160 mmHg or diastolic blood pressure &gt; 100&#xD;
             mmHg&#xD;
&#xD;
          -  Prior history of cardiovascular events (like stroke, transient ischemic attack,&#xD;
             myocardial infarction, angina, heart failure...)&#xD;
&#xD;
          -  Prior history of cancer within the last 3 years&#xD;
&#xD;
          -  Thyroid disease - untreated&#xD;
&#xD;
          -  Type 1 or 2 diabetes or fasting glucose &gt; 7.0 mmol/L&#xD;
&#xD;
          -  Claustrophobia&#xD;
&#xD;
          -  Anemia - Hb &lt; 120 g/L&#xD;
&#xD;
          -  Creatinine &gt; 100 μmol/L&#xD;
&#xD;
          -  Hepatic dysfunction - AST/ALT &gt; 3 times normal limit&#xD;
&#xD;
          -  Blood coagulation problems (i.e. bleeding predisposition)&#xD;
&#xD;
          -  Autoimmune diseases&#xD;
&#xD;
          -  Chronic inflammatory diseases&#xD;
&#xD;
          -  Concomitant medications&#xD;
&#xD;
               -  Hormone replacement therapy (except thyroid hormone at a stable dose)&#xD;
&#xD;
               -  Systemic corticosteroids&#xD;
&#xD;
               -  Anti-psychotic medications - psycho-active medication&#xD;
&#xD;
               -  Anticoagulant treatment (Aspirin, NSAIDs, warfarin, coumadin..)&#xD;
&#xD;
               -  Adrenergic agonist&#xD;
&#xD;
               -  Anti-hypertensive&#xD;
&#xD;
               -  Weight-loss&#xD;
&#xD;
          -  Known substance abuse&#xD;
&#xD;
          -  Allergy to seafood or fish&#xD;
&#xD;
          -  Cancellation of the same scheduled testing visit, twice&#xD;
&#xD;
          -  Lack of time to participate in the full length of the study (18 weeks)&#xD;
&#xD;
          -  Have exceeded the annual total allowed radiation dose (like X-ray scans and/or&#xD;
             tomography in the previous year or in the year to come) according to the physician's&#xD;
             judgement.&#xD;
&#xD;
          -  All other medical or psychological conditions deemed inappropriate according to the&#xD;
             physician&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>45 Years</minimum_age>
    <maximum_age>74 Years</maximum_age>
    <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>May Faraj, PDt, PhD</last_name>
    <role>Principal Investigator</role>
    <affiliation>Montreal Clinical Research Institute/ University of Montreal</affiliation>
  </overall_official>
  <reference>
    <citation>Lamantia V, Bissonnette S, Wassef H, Cyr Y, Baass A, Dufour R, Rabasa-Lhoret R, Faraj M. ApoB-lipoproteins and dysfunctional white adipose tissue: Relation to risk factors for type 2 diabetes in humans. J Clin Lipidol. 2017 Jan-Feb;11(1):34-45.e2. doi: 10.1016/j.jacl.2016.09.013. Epub 2016 Oct 3.</citation>
    <PMID>28391908</PMID>
  </reference>
  <reference>
    <citation>Bissonnette S, Saint-Pierre N, Lamantia V, Cyr Y, Wassef H, Faraj M. Plasma IL-1Ra: linking hyperapoB to risk factors for type 2 diabetes independent of obesity in humans. Nutr Diabetes. 2015 Sep 28;5(9):e180. doi: 10.1038/nutd.2015.30.</citation>
    <PMID>26417659</PMID>
  </reference>
  <reference>
    <citation>Lamantia V, Bissonnette S, Provost V, Devaux M, Cyr Y, Daneault C, Rosiers CD, Faraj M. The Association of Polyunsaturated Fatty Acid delta-5-Desaturase Activity with Risk Factors for Type 2 Diabetes Is Dependent on Plasma ApoB-Lipoproteins in Overweight and Obese Adults. J Nutr. 2019 Jan 1;149(1):57-67. doi: 10.1093/jn/nxy238.</citation>
    <PMID>30535058</PMID>
  </reference>
  <reference>
    <citation>Skeldon AM, Faraj M, Saleh M. Caspases and inflammasomes in metabolic inflammation. Immunol Cell Biol. 2014 Apr;92(4):304-13. doi: 10.1038/icb.2014.5. Epub 2014 Feb 11.</citation>
    <PMID>24518981</PMID>
  </reference>
  <reference>
    <citation>Lamantia V, Sniderman A, Faraj M. Nutritional management of hyperapoB. Nutr Res Rev. 2016 Dec;29(2):202-233. doi: 10.1017/S0954422416000147. Epub 2016 Nov 8.</citation>
    <PMID>27821191</PMID>
  </reference>
  <reference>
    <citation>Faraj M. LDL, LDL receptors, and PCSK9 as modulators of the risk for type 2 diabetes: a focus on white adipose tissue. J Biomed Res. 2020 Mar 12;34(4):251-259. doi: 10.7555/JBR.34.20190124.</citation>
    <PMID>32701068</PMID>
  </reference>
  <results_reference>
    <citation>Simon Bissonnette, Valerie Lamantia, Yannick Cyr, Viviane Provost, Marie Devaux, May Faraj. Native LDL are Priming Signals of White Adipose Tissue NLRP3 Inflammasome in Overweight and Obese Subjects. Atherosclerosis Supplements (Abstract). 2018; 32:123</citation>
  </results_reference>
  <verification_date>August 2020</verification_date>
  <study_first_submitted>July 29, 2020</study_first_submitted>
  <study_first_submitted_qc>July 29, 2020</study_first_submitted_qc>
  <study_first_posted type="Actual">August 3, 2020</study_first_posted>
  <last_update_submitted>August 3, 2020</last_update_submitted>
  <last_update_submitted_qc>August 3, 2020</last_update_submitted_qc>
  <last_update_posted type="Actual">August 5, 2020</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor-Investigator</responsible_party_type>
    <investigator_affiliation>Institut de Recherches Cliniques de Montreal</investigator_affiliation>
    <investigator_full_name>May Faraj, PDt, PhD</investigator_full_name>
    <investigator_title>Professor</investigator_title>
  </responsible_party>
  <keyword>HyperapoB</keyword>
  <keyword>ApoB-lipoproteins</keyword>
  <keyword>White adipose tissue</keyword>
  <keyword>NLRP3 inflammasome</keyword>
  <keyword>Fat metabolism</keyword>
  <keyword>Insulin sensitivity and secretion</keyword>
  <keyword>Systemic inflammation</keyword>
  <keyword>Eicosapentaenoic acid (EPA)</keyword>
  <keyword>Docosahexaenoic acid (DHA)</keyword>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Insulin Resistance</mesh_term>
    <mesh_term>Hypersensitivity</mesh_term>
  </condition_browse>
  <patient_data>
    <sharing_ipd>No</sharing_ipd>
    <ipd_description>Biological samples (plasma and adipose tissue) can be made available for analysis by other investigators. However data statistical analysis incorporating complete IPD must be conducted by the research team of Dr May Faraj as per subject consent form.</ipd_description>
  </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/NCT04496154
org study id: 2013-14 and 2014-756
secondary id: NUT273741
nct id: NCT04496154
lead sponsor: 
collaborator: 
has dmc: No
is fda regulated drug: No
is fda regulated device: No
In this project, investigators explored the role of the particles that carry "bad
      cholesterol" in the blood (termed LDL) that are known to promote heart disease, in the
      promotion of type 2 diabetes (T2D) in humans. In specific, they investigated how these
      particles may induce the activation of an immune pathway in human fat tissue leading to
      multiple anomalies that favors T2D. They also explored whether omega-3 fatty acids, which are
      the type of fat found in fish oils can counterbalance the negative effects of LDL in fat
      tissue, thus providing a natural way to help reduce the risk for T2D in subjects with
      elevated blood LDL.

      To do so, 41 subjects who were free of disease or medication affecting metabolism were
      enrolled at the Montreal Clinical Research Institute between 2013 and 2019 and were placed on
      an intervention with omega-3 fatty acids supplementation for 12 weeks (2.7 g/day, Triple
      Strength Omega-3 from Webbers Naturals). Investigators examined the effects of LDL and
      omega-3 on risk factors for T2D before and after the intervention in the whole body and
      specifically in fat tissue biopsies taken from the hip region. Eighty percent of the subjects
      who were enrolled into the study completed the intervention.
Diabetes-attributed deaths, mostly type 2 diabetes (T2D), total more than 40,000 per year,
      out of which 80% are secondary to cardiovascular disease and stroke. Research from the
      investigators' lab and others suggest that elevated atherogenic apoB-lipoproteins may not be
      a mere consequence of T2D but also a cause. They reported that high number of
      apoB-lipoproteins (apoB) may directly induce subcutaneous white adipose tissue dysfunction
      and related risk factors for T2D in humans; however underlying mechanisms were yet unclear.

      Strong evidence implicates a specific innate immunity complex, the NLRP3 inflammasome (NLRP3
      for Nucleotide-binding domain and Leucine-rich repeat Receptor, containing a Pyrin domain 3)
      in white adipose tissue dysfunction and associated metabolic anomalies in mice and humans.
      Preliminary evidence from the investigators lab and their collaborator (Dr Maya Saleh, at
      McGill University) indicated that native apoB-lipoproteins activate the NLRP3 inflammasome in
      murine bone marrow derived macrophages. On the other hand, omega-3 fatty acids were reported
      to inhibit the NLRP3 inflammasome in immune cells.

      Thus the hypotheses that were examined in this study were that:

      Primary hypothesis:

        1. Subjects with high plasma apoB have higher NLRP3 inflammasome activity in white adipose
           tissue indicated by higher interleukin 1 beta (IL-1β) secretion.

           Higher white adipose tissue IL-1β secretion was expected to be associated with risk
           factors for T2D documented in subjects with high plasma apoB, namely white adipose
           tissue dysfunction, systemic inflammation, postprandial hypertriglyceridemia, insulin
           resistance and hyperinsulinemia

           Secondary hypotheses:

        2. Twelve-week supplementation with omega-3 fatty acids (2.7 g/day, eicosapentaenoic acid
           (EPA) and docosahexaenoic (DHA), 2:1) induces a greater reduction in white adipose
           tissue NLRP3 inflammasome activity and related risk factors for T2D in subjects with
           high plasma apoB.

        3. Subject native LDL (representing > 90% of apoB-lipoproteins) directly prime and/or
           activate the NLRP3 inflammasome in subject own white adipose tissue ex vivo, while
           EPA/DHA inhibit it.

      Forty-one subjects (34% men) were enrolled in the study, of whom 33 subjects completed the
      12-week omega-3 intervention (drop out/exclusion rate = 20%). Baseline and post-intervention
      risk factors for T2D measured in these subjects were; white adipose tissue and systemic NLRP3
      inflammasome activity, dietary fat clearance (after a high fat meal), adipose tissue function
      (ex vivo after a subcutaneous adipose tissue needle biopsy), and insulin secretion and
      sensitivity (by gold-standard Botnia clamp technique).
allocation: N/A
intervention model: Single Group Assignment
primary purpose: Prevention
masking: None (Open Label)
measure: Fasting white adipose tissue NLRP3 inflammasome activation
time frame: Baseline
description: White adipose tissue medium accumulation of interleukin 1 beta (IL-1β) ex vivo over 4 hours (pg/mg tissue by AlphaLISA)
measure: Fasting white adipose tissue NLRP3 inflammasome activation
time frame: Change at 12 weeks from baseline
description: White adipose tissue medium accumulation of interleukin 1 beta (IL-1β) ex vivo over 4 hours (pg/mg tissue by AlphaLISA)
measure: White adipose tissue inflammation profile
time frame: Baseline
description: Fasting and 4 hour-postprandial change in NLRP3 inflammasome related inflammatory parameters; including protein and gene expression of IL1B and NLRP3 (by immunohistochemistry and RT-PCR), secretion of IL-1β (per mg tissue by AlphaLISA) and macrophage infiltration of white adipose tissue (by RT-PCR)
measure: White adipose tissue inflammation profile
time frame: Change at 12 weeks from baseline
description: Fasting and 4 hour-postprandial change in NLRP3 inflammasome related inflammatory parameters; including protein and gene expression of IL1B and NLRP3 (by immunohistochemistry and RT-PCR), secretion of IL-1β (per mg tissue by AlphaLISA) and macrophage infiltration of white adipose tissue (by RT-PCR)
measure: White adipose tissue function ex vivo
time frame: Baseline
description: Fasting in situ lipoprotein lipase activity (nmol 3H-triglyceride/mg tissue)
measure: White adipose tissue function ex vivo
time frame: Change at 12 weeks from baseline
description: Fasting in situ lipoprotein lipase activity (nmol 3H-triglyceride/mg tissue)
measure: Postprandial fat metabolism
time frame: Baseline
description: Area under the 6 hour time curve of plasma triglycerides (mmol/hour) after a high-fat meal (66% fat)
measure: Postprandial fat metabolism
time frame: Change at 12 weeks from baseline
description: Area under the 6 hour time curve of plasma triglycerides (mmol/hour) after a high-fat meal (66% fat)
measure: Systemic inflammation
time frame: Baseline
description: Fasting plasma inflammatory parameters including IL-1Ra and IL-1β (pg/mL by AlphaLISA)
measure: Systemic inflammation
time frame: Change at 12 weeks from baseline
description: Fasting plasma inflammatory parameters including IL-1Ra and IL-1β (pg/mL by AlphaLISA)
measure: Insulin sensitivity and secretion
time frame: Baseline
description: Glucose-induced insulin secretion (uU/mL/min) and insulin sensitivity (glucose infusion rate mg/kg/min) measured by Botnia clamp
measure: Insulin sensitivity and secretion
time frame: Change at 12 weeks from baseline
description: Glucose-induced insulin secretion (uU/mL/min) and insulin sensitivity (glucose infusion rate mg/kg/min) measured by Botnia clamp
measure: Fatty acid profile in the phospholipid fraction of plasma and red blood cells
time frame: Baseline
description: (As μmol/L by gas chromatography mass spectrometry)
measure: Fatty acid profile in the phospholipid fraction of plasma and red blood cells
time frame: Change at 12 weeks from baseline
description: (As μmol/L by gas chromatography mass spectrometry)
measure: Body composition
time frame: Baseline
description: Fat and lean body mass (as kg by dual energy x-ray absorptiometry)
measure: Body composition
time frame: Change at 12 weeks from baseline
description: Fat and lean body mass (as kg by dual energy x-ray absorptiometry)
measure: Energy expenditure
time frame: Baseline
description: Fasting and postprandial during the 6-hour high fat meal challenge (as kcal/hr using indirect calorimetry)
measure: Energy expenditure
time frame: Change at 12 weeks from baseline
description: Fasting and postprandial during the 6-hour high fat meal challenge (as kcal/hr using indirect calorimetry)
measure: Energy intake
time frame: Baseline
description: (Average of 3 day energy intake as kcal/day collected by 3-day dietary records)
measure: Energy intake
time frame: Change at 12 weeks from baseline
description: (Average of 3 day energy intake as kcal/day collected by 3-day dietary records)
measure: Post-hoc analyses of white adipose tissue receptors for apoB-lipoproteins
time frame: Baseline
description: Fasting and 4 hour-postprandial change in white adipose tissue surface-expression LDLR and CD36 (% of control by immunohistochemistry in white adipose tissue slides)
measure: Post-hoc analyses of white adipose tissue receptors for apoB-lipoproteins
time frame: Change at 12 weeks from baseline
description: Fasting and 4 hour-postprandial change in white adipose tissue surface-expression LDLR and CD36 (% of control by immunohistochemistry in white adipose tissue slides)
measure: Post-hoc analyses of white adipose tissue genes related to lipid and lipoprotein metabolism
time frame: Baseline
description: Fasting and 4-hour postprandial change in gene expression including LDLR, CD36, SREBP1, SREBP2 and HMGCR (by RT-PCR)
measure: Post-hoc analyses of white adipose tissue genes related to lipid and lipoprotein metabolism
time frame: Change at 12 weeks from baseline
description: Fasting and 4-hour postprandial change in gene expression including LDLR, CD36, SREBP1, SREBP2 and HMGCR (by RT-PCR)
arm group label: Omega-3 fatty acids
arm group type: Experimental
description: 3 oral softgels (600 mg EPA and 300 mg DHA / softgel), Triple Strength Omega-3 from Webber Naturals
intervention type: Dietary Supplement
intervention name: Omega-3 fatty acids (2.7 g/d, EPA:DHA, 2:1)
description: Triple Strength Omega-3 from Webber Naturals
arm group label: Omega-3 fatty acids
criteria: 
gender: All
minimum age: 45 Years
maximum age: 74 Years
healthy volunteers: Accepts Healthy Volunteers
last name: May Faraj, PDt, PhD
role: Principal Investigator
affiliation: Montreal Clinical Research Institute/ University of Montreal
citation: Lamantia V, Bissonnette S, Wassef H, Cyr Y, Baass A, Dufour R, Rabasa-Lhoret R, Faraj M. ApoB-lipoproteins and dysfunctional white adipose tissue: Relation to risk factors for type 2 diabetes in humans. J Clin Lipidol. 2017 Jan-Feb;11(1):34-45.e2. doi: 10.1016/j.jacl.2016.09.013. Epub 2016 Oct 3.
PMID: 28391908
citation: Bissonnette S, Saint-Pierre N, Lamantia V, Cyr Y, Wassef H, Faraj M. Plasma IL-1Ra: linking hyperapoB to risk factors for type 2 diabetes independent of obesity in humans. Nutr Diabetes. 2015 Sep 28;5(9):e180. doi: 10.1038/nutd.2015.30.
PMID: 26417659
citation: Lamantia V, Bissonnette S, Provost V, Devaux M, Cyr Y, Daneault C, Rosiers CD, Faraj M. The Association of Polyunsaturated Fatty Acid delta-5-Desaturase Activity with Risk Factors for Type 2 Diabetes Is Dependent on Plasma ApoB-Lipoproteins in Overweight and Obese Adults. J Nutr. 2019 Jan 1;149(1):57-67. doi: 10.1093/jn/nxy238.
PMID: 30535058
citation: Skeldon AM, Faraj M, Saleh M. Caspases and inflammasomes in metabolic inflammation. Immunol Cell Biol. 2014 Apr;92(4):304-13. doi: 10.1038/icb.2014.5. Epub 2014 Feb 11.
PMID: 24518981
citation: Lamantia V, Sniderman A, Faraj M. Nutritional management of hyperapoB. Nutr Res Rev. 2016 Dec;29(2):202-233. doi: 10.1017/S0954422416000147. Epub 2016 Nov 8.
PMID: 27821191
citation: Faraj M. LDL, LDL receptors, and PCSK9 as modulators of the risk for type 2 diabetes: a focus on white adipose tissue. J Biomed Res. 2020 Mar 12;34(4):251-259. doi: 10.7555/JBR.34.20190124.
PMID: 32701068
citation: Simon Bissonnette, Valerie Lamantia, Yannick Cyr, Viviane Provost, Marie Devaux, May Faraj. Native LDL are Priming Signals of White Adipose Tissue NLRP3 Inflammasome in Overweight and Obese Subjects. Atherosclerosis Supplements (Abstract). 2018; 32:123
responsible party type: Sponsor-Investigator
investigator affiliation: Institut de Recherches Cliniques de Montreal
investigator full name: May Faraj, PDt, PhD
investigator title: Professor
mesh term: Insulin Resistance
mesh term: Hypersensitivity
sharing ipd: No
ipd description: Biological samples (plasma and adipose tissue) can be made available for analysis by other investigators. However data statistical analysis incorporating complete IPD must be conducted by the research team of Dr May Faraj as per subject consent form.