Vitamin D and Adipose Tissue Inflammation

This study has been completed.
Sponsor:
Collaborator:
University of Washington
Information provided by (Responsible Party):
Kratz, Mario, Fred Hutchinson Cancer Research Center
ClinicalTrials.gov Identifier:
NCT01477034
First received: November 1, 2011
Last updated: April 2, 2014
Last verified: April 2014
  Purpose

Chronic, low-grade adipose tissue inflammation is a major risk factor for type 2 diabetes mellitus. The cause of adipose tissue inflammation has remained largely unclear. We hypothesize that vitamin D deficiency predisposes individuals to the development of adipose tissue inflammation, and that treatment of vitamin D deficient subjects with high dose vitamin D will reduce adipose tissue inflammation.


Condition Intervention
Vitamin D Deficiency
Obesity
Type 2 Diabetes Mellitus
Intestinal Permeability
Dietary Supplement: Vitamin D3

Study Type: Interventional
Study Design: Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Double Blind (Subject, Investigator, Outcomes Assessor)
Primary Purpose: Basic Science
Official Title: Vitamin D and Adipose Tissue Inflammation

Resource links provided by NLM:


Further study details as provided by Fred Hutchinson Cancer Research Center:

Primary Outcome Measures:
  • Tumor Necrosis Factor alpha expression in adipose tissue [ Time Frame: Change from baseline to the 6 month visit ] [ Designated as safety issue: No ]
    Total RNA will be extracted from whole adipose tissue. TNF alpha mRNA will be quantified using PCR, and normalized using a normalization factor based on three housekeeping genes. We will compute the change in adipose tissue TNF alpha mRNA level between baseline and the 6 month visit.

  • Tumor Necrosis Factor alpha expression in adipose tissue [ Time Frame: Change from baseline to the 3 month visit ] [ Designated as safety issue: No ]
    Total RNA will be extracted from whole adipose tissue. TNF alpha mRNA will be quantified using PCR, and normalized using a normalization factor based on three housekeeping genes. We will compute the change in adipose tissue TNF alpha mRNA level between baseline and the 3 month visit.


Secondary Outcome Measures:
  • Plasma concentrations of 24,25-dihydroxy vitamin D [24,25(OH)2D] [ Time Frame: Change from baseline to the 6 month visit ] [ Designated as safety issue: No ]
    The concentration of 24,25(OH)2D will be measured in fasting plasma using high performance liquid chromatography-tandem mass spectometry (LC/MS/MS).

  • Adipose tissue concentration of 25-hydroxy vitamin D [25(OH)D] [ Time Frame: Change from baseline to the 6 month visit ] [ Designated as safety issue: No ]
    Adipose tissue 25(OH)D will be measured using high performance liquid chromatography-tandem mass spectometry (LC/MS/MS.)

  • CD16+ macrophages in adipose tissue [ Time Frame: Change from baseline to the 6 month visit ] [ Designated as safety issue: No ]
    The number or CD16+ macrophages in adipose tissue, normalized to the total number of CD14+CD206+ macrophages or the total number of CD45+ cells, will be measured using multi-parameter flow cytometry.

  • CD8+ T cells in adipose tissue [ Time Frame: Change from baseline to the 6 month visit ] [ Designated as safety issue: No ]
    The number of CD8+ T cells in adipose tissue, normalized to the total number of CD3+ cells, will be measured using multi-parameter flow cytometry.

  • Plasma concentration of 25-hydroxy vitamin D [25(OH)D] [ Time Frame: Change from baseline to the 6 month visit ] [ Designated as safety issue: No ]
    Plasma 25(OH)D will be measured by high performance liquid chromatography-tandem mass spectometry (LC/MS/MS).

  • Adipose tissue concentration of cholecalciferol (vitamin D3) [ Time Frame: Change from baseline to the 6 month visit ] [ Designated as safety issue: No ]
    Adipose tissue concentrations of cholecalciferol will be measured by high performance liquid chromatography-tandem mass spectometry (LC/MS/MS)

  • CD11c+ macrophages in adipose tissue [ Time Frame: Change from baseline to the 6 month visit ] [ Designated as safety issue: No ]
    The number of CD11c+ macrophages in adipose tissue, normalized to the total number of CD45+ cells, will be measured by multi-parameter flow cytometry.

  • CD4+CD25+ T cells in adipose tissue [ Time Frame: Change from baseline to the 6 month visit ] [ Designated as safety issue: No ]
    The number of CD4+CD25+ T cells in adipose tissue, normalized to the total number of CD4+ T cells, will be measured by multi-parameter flow cytometry.

  • Intestinal permeability, as assessed by the 5-hour urinary lactulose/mannitol test [ Time Frame: Change from baseline to 6 month clinic visit. ] [ Designated as safety issue: No ]
    Intestinal permeability will be assessed at each clinic visit by administering 2g of mannitol and 5 g of lactulose to the oral glucose tolerance test beverage followed by collection of urine for 5 hours afterwards. Recovery of mannitol and lactulose in urine will be measured by gas chromatography, and will be indicative of the degree of intestinal permeability.

  • Fasting plasma zonulin concentrations [ Time Frame: Change from baseline to 6 month clinic visit ] [ Designated as safety issue: No ]
    Zonulin concentrations will be measured by enzyme-linked immunosorbent assay in fasting plasma collected at all clinic visits. Plasma zonulin is a marker of intestinal permeability.

  • Fasting plasma lipopolysaccharide binding protein (LBP) [ Time Frame: Change from baseline to 6 month clinic visit ] [ Designated as safety issue: No ]
    Lipopolysaccharide binding protein (LBP) will be measured by enzyme-linked immunosorbent assay in fasting plasma collected at all clinic visits. LBP is an acute phase protein secreted by the liver in response to endotoxin (lipopolysaccharide) exposure.

  • Plasma concentrations of 24,25-dihydroxy vitamin D [24,25(OH)2D] [ Time Frame: Change from baseline to the 3 month visit ] [ Designated as safety issue: No ]
    The concentration of 24,25(OH)2D will be measured in fasting plasma using high performance liquid chromatography-tandem mass spectometry (LC/MS/MS).

  • Adipose tissue concentration of 25-hydroxy vitamin D [25(OH)D] [ Time Frame: Change from baseline to the 3 month visit ] [ Designated as safety issue: No ]
    Adipose tissue 25(OH)D will be measured using high performance liquid chromatography-tandem mass spectometry (LC/MS/MS.)

  • CD16+ macrophages in adipose tissue [ Time Frame: Change from baseline to the 3 month visit ] [ Designated as safety issue: No ]
    The number or CD16+ macrophages in adipose tissue, normalized to the total number of CD14+CD206+ macrophages or the total number of CD45+ cells, will be measured using multi-parameter flow cytometry.

  • CD8+ T cells in adipose tissue [ Time Frame: Change from baseline to the 3 month visit ] [ Designated as safety issue: No ]
    The number of CD8+ T cells in adipose tissue, normalized to the total number of CD3+ cells, will be measured using multi-parameter flow cytometry.

  • Plasma concentration of 25-hydroxy vitamin D [25(OH)D] [ Time Frame: Change from baseline to the 3 month visit ] [ Designated as safety issue: No ]
    Plasma 25(OH)D will be measured by high performance liquid chromatography-tandem mass spectometry (LC/MS/MS).

  • Adipose tissue concentration of cholecalciferol (vitamin D3) [ Time Frame: Change from baseline to the 3 month visit ] [ Designated as safety issue: No ]
    Adipose tissue concentrations of cholecalciferol will be measured by high performance liquid chromatography-tandem mass spectometry (LC/MS/MS)

  • CD11c+ macrophages in adipose tissue [ Time Frame: Change from baseline to the 3 month visit ] [ Designated as safety issue: No ]
    The number of CD11c+ macrophages in adipose tissue, normalized to the total number of CD45+ cells, will be measured by multi-parameter flow cytometry.

  • CD4+CD25+ T cells in adipose tissue [ Time Frame: Change from baseline to the 3 month visit ] [ Designated as safety issue: No ]
    The number of CD4+CD25+ T cells in adipose tissue, normalized to the total number of CD4+ T cells, will be measured by multi-parameter flow cytometry.

  • Intestinal permeability, as assessed by the 5-hour urinary lactulose/mannitol test [ Time Frame: Change from baseline to 3 month clinic visit. ] [ Designated as safety issue: No ]
    Intestinal permeability will be assessed at each clinic visit by administering 2g of mannitol and 5 g of lactulose to the oral glucose tolerance test beverage followed by collection of urine for 5 hours afterwards. Recovery of mannitol and lactulose in urine will be measured by gas chromatography, and will be indicative of the degree of intestinal permeability.

  • Fasting plasma zonulin concentrations [ Time Frame: Change from baseline to 3 month clinic visit ] [ Designated as safety issue: No ]
    Zonulin concentrations will be measured by enzyme-linked immunosorbent assay in fasting plasma collected at all clinic visits. Plasma zonulin is a marker of intestinal permeability.

  • Fasting plasma lipopolysaccharide binding protein (LBP) [ Time Frame: Change from baseline to 3 month clinic visit ] [ Designated as safety issue: No ]
    Lipopolysaccharide binding protein (LBP) will be measured by enzyme-linked immunosorbent assay in fasting plasma collected at all clinic visits. LBP is an acute phase protein secreted by the liver in response to endotoxin (lipopolysaccharide) exposure.


Enrollment: 18
Study Start Date: November 2011
Study Completion Date: June 2013
Primary Completion Date: June 2013 (Final data collection date for primary outcome measure)
Arms Assigned Interventions
Experimental: 2,000 IU/day vitamin D3 x 6 months
Subjects will take a 2,000 IU daily vitamin D3 supplement for 6 months.
Dietary Supplement: Vitamin D3
2,000 or 4,000 IU/day vitamin D3 for 3 or 6 months.
Other Name: Carlson Labs Vitamin D3 capsules (2,000/4,000 IU/capsule)
Experimental: 4,000 IU/day vitamin D3 x 6 months
Subjects will take a 4,000 IU daily vitamin D3 supplement for 6 months.
Dietary Supplement: Vitamin D3
2,000 or 4,000 IU/day vitamin D3 for 3 or 6 months.
Other Name: Carlson Labs Vitamin D3 capsules (2,000/4,000 IU/capsule)
Experimental: 2,000 IU/day vitamin D3 x 3 months
Subjects will take a 2,000 IU daily vitamin D3 supplement for 3 months.
Dietary Supplement: Vitamin D3
2,000 or 4,000 IU/day vitamin D3 for 3 or 6 months.
Other Name: Carlson Labs Vitamin D3 capsules (2,000/4,000 IU/capsule)
Experimental: 4,000 IU/day vitamin D3 x 3 months
Subjects will take a 4,000 IU daily vitamin D3 supplement for 3 months.
Dietary Supplement: Vitamin D3
2,000 or 4,000 IU/day vitamin D3 for 3 or 6 months.
Other Name: Carlson Labs Vitamin D3 capsules (2,000/4,000 IU/capsule)

Detailed Description:

The objective of this project is to investigate whether vitamin D modulates chronic low-grade adipose tissue inflammation in overweight and obese, vitamin D deficient men and women.

Obesity is associated with insulin resistance and an increased risk for type 2 diabetes mellitus. Numerous studies, mostly conducted in mouse models of obesity, strongly suggest that chronic low-grade inflammation of adipose and other tissues is the major mechanism by which increased adiposity is linked to insulin resistance. Adipose tissue inflammation may therefore be a promising therapeutic target to reduce insulin resistance and the risk of type 2 diabetes mellitus in obese individuals.

Based on several lines of evidence, we hypothesize that vitamin D is an environmental factor that affects the course of the inflammatory response in most tissues of the body, including adipose tissue. In our previous studies, we found that circulating plasma concentrations of 25-hydroxy vitamin D (25-OH-D) and the primary degradation product 24,25-dihydroxy vitamin D (24,25-OH2-D) were significantly associated with adipose tissue expression of adiponectin and negatively with TNF-alpha, even when adjusted for body mass index. Because these previous studies were cross-sectional, it is critical to complete an intervention study in humans to determine whether the observed association of vitamin D levels and adipose tissue inflammation is causal. The objectives of this pilot study are therefore to collect relevant preliminary data, and to begin an exploration of the mechanisms underlying this association such as intestinal permeability.

Increased intestinal permeability may contribute to chronic low-grade inflammation and signaling through the vitamin D receptor plays an important role in the maintenance of intestinal integrity. We will assess whether normalization of vitamin D status is associated with changes in intestinal permeability.

  Eligibility

Ages Eligible for Study:   18 Years to 65 Years
Genders Eligible for Study:   Both
Accepts Healthy Volunteers:   Yes
Criteria

Inclusion Criteria:

  • Age: 18-65 years;
  • BMI ≥25 kg/m2;
  • Plasma 25-OH-vitamin D between 7 and 20 ng/mL
  • Weight stable to within 10 pounds for 6 months prior to entering the study, and within 30 pounds of their lifetime maximum weight (excluding pregnancy);
  • Ability to be admitted for ~6.5 hours on three occasions to the FHCRC Prevention Center,
  • Ability to provide informed written consent;
  • Willingness to take vitamin D3 capsules daily for 6 months

Exclusion Criteria:

  • Chronic disease such as thyroid disease, liver disease, or kidney disease;
  • Diabetes mellitus, or fasting glucose >125 mg/dL;
  • Chronic inflammatory condition such as autoimmune disease or inflammatory bowel disease;
  • Malabsorption syndromes (untreated celiac disease; condition after stomach or intestinal resection);
  • Current or recent (within one month) chronic intake of medications likely to interfere with study endpoints [(insulin, antidiabetics, anabolic steroids, glucocorticosteroids, statins, blood thinners (warfarin, aspirin), non-steroidal anti-inflammatory drugs (if daily)];
  • Current or recent (within 3 months) intake of vitamin D in excess of 600 IU/day;
  • Anemia, recent history (within 3 months) of anemia; recent (within 3 months) blood donation; recent (within 3 months) participation in another study that involved blood draws; or plans to participate in other research that involves blood draws during the study period;
  • Pregnancy in the last 6 months, plans to become pregnant during the study period, or current breastfeeding.
  Contacts and Locations
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Please refer to this study by its ClinicalTrials.gov identifier: NCT01477034

Locations
United States, Washington
Fred Hutchinson Cancer Research Center
Seattle, Washington, United States, 98109
Sponsors and Collaborators
Fred Hutchinson Cancer Research Center
University of Washington
Investigators
Principal Investigator: Mario Kratz, Ph.D. Fred Hutchinson Cancer Research Center
  More Information

No publications provided

Responsible Party: Kratz, Mario, Assistant Member, Fred Hutchinson Cancer Research Center
ClinicalTrials.gov Identifier: NCT01477034     History of Changes
Other Study ID Numbers: UW NORC P&F KRATZ, 7598
Study First Received: November 1, 2011
Last Updated: April 2, 2014
Health Authority: United States: Institutional Review Board

Keywords provided by Fred Hutchinson Cancer Research Center:
Vitamin D deficiency
Obesity
Inflammation
Low-grade, chronic inflammation
Adipose tissue inflammation
Diabetes
Type 2 diabetes mellitus
Insulin resistance
Intestinal permeability

Additional relevant MeSH terms:
Diabetes Mellitus
Diabetes Mellitus, Type 2
Obesity
Inflammation
Vitamin D Deficiency
Glucose Metabolism Disorders
Metabolic Diseases
Endocrine System Diseases
Overnutrition
Nutrition Disorders
Overweight
Body Weight
Signs and Symptoms
Pathologic Processes
Avitaminosis
Deficiency Diseases
Malnutrition
Vitamins
Vitamin D
Ergocalciferols
Cholecalciferol
Micronutrients
Growth Substances
Physiological Effects of Drugs
Pharmacologic Actions
Bone Density Conservation Agents

ClinicalTrials.gov processed this record on October 19, 2014