Effect of Vitamin D Repletion on Insulin Resistance and Systemic Inflammation

• Sponsor: Albert Einstein College of Medicine (Other)
• Overall Status: Completed
• CT.gov ID: NCT01354964
• Collaborator: National Center for Research Resources (NCRR) (NIH)
• Enrollment: 19
• Location: 1
• Arms: 2
• Actual Duration (Months): 77.7
• Patients Per Site Per Month: 0.2

Study Details

Study Description

Brief Summary

The purpose of this research is to study the effects of Vitamin D supplementation on the body's response to insulin (a hormone that controls blood sugar), on inflammation, and on specific cells and processes in fat tissue.

Condition or Disease	Intervention/Treatment	Phase
Insulin Resistance	Drug: Vitamin D; Drug: Placebo	Phase 2

Detailed Description

Over the last several years, studies have shown that low vitamin D levels may increase risk of developing Type 2 Diabetes. The investigators will administer vitamin D3 (cholecalciferol) to non-diabetic, insulin resistant subjects with vitamin D deficiency (total vitamin D levels <20 ng/ml) to increase the level of vitamin D3. The investigators will study the effects of increased Vitamin D on insulin action, adipose tissue inflammation, and on certain cells and processes in fat tissue.

Investigators will study participants with a procedure called a "pancreatic clamp" study. During the clamp procedure, glucose (a sugar) and insulin (a hormone produced in the pancreas that regulates the amount of glucose in the blood) are infused with an intravenous catheter, and blood samples are collected periodically throughout the procedure to measure blood sugar levels and the levels of several hormones that are found in the body and are related to glucose metabolism. Adipose tissue inflammation will be measured using the following inflammatory markers: IL-6, PAI-1, TNF-alpha, and iNOS.

Study Design

• Study Type: Interventional
• Actual Enrollment: 19 participants
• Allocation: Randomized
• Intervention Model: Parallel Assignment
• Masking: Double (Participant, Investigator)
• Primary Purpose: Treatment
• Official Title: Effect of Vitamin D Repletion on Insulin Resistance and Systemic Inflammation
• Actual Study Start Date: Mar 13, 2009
• Actual Primary Completion Date: Jun 3, 2015
• Actual Study Completion Date: Sep 3, 2015

Arms and Interventions

Arm	Intervention/Treatment
Experimental: Vitamin D Participants received weekly oral vitamin D drops using a weight-based calculated dosage for up to six months.	Drug: Vitamin D; Other Names: Vitamin D3 (cholecalciferol)
Placebo Comparator: Placebo Participants received weekly oral placebo drops (similar in taste and appearance to vitamin D) for up to six months.	Drug: Placebo

Outcome Measures

Primary Outcome Measures

1. Percent Change in Hepatic Insulin Sensitivity [2nd clamp visit (after up to 3 months) and 3rd clamp visit (after up to 6 months)]

   Endogenous glucose production (EGP) was assessed at each study visit to evaluate hepatic insulin sensitivity. Percent change between the EGP at baseline and second visit (after treatment for up to 3 months with Vitamin D to reach a target level of ≥30 ng/ml), and baseline and third visits (after treatment for up to 6 months with Vitamin D in order to reach a target level of ≥50 ng/ml) will be calculated.

Secondary Outcome Measures

1. Percent Change in Peripheral Glucose Uptake [2nd clamp visit (up to 3 months) and 3rd clamp visit (up to 6 months)]

   The rate of glucose uptake to determine peripheral insulin sensitivity was measured using the rate of disappearance (Rd) of glucose at each study visit. Percent change between the Rd at baseline and second visit (after treatment with Vitamin D for up to 3 months to target level of ≥30 ng/ml), and baseline and third visits (after treatment with Vitamin D for up to 6 months to target level of ≥50 ng/ml) will be calculated.

2. Evaluated Expression of Pro-inflammatory Gene TNF-α [2nd clamp visit (up to 3 months) and 3rd clamp visit (up to 6 months)]

   Adipose tissue macrophages will be isolated from subcutaneous abdominal adipose tissue, and will be quantified by fluorescence activated cell sorting (FACS) analysis. TNF-α gene expression will be examined by real-time (rt-PCR) and will provide a measure of macrophage activation at baseline, at 2nd study visit (after treatment with Vitamin D to a goal level of ≥30 ng/ml), and at 3rd study visit (goal Vitamin D level of ≥50 ng/ml). The mRNA copy number is then compared with a reference gene copy number (5 commonly used house keeping genes [HKGs]) as a ratio, which is a measure of relative gene expression.

3. Evaluated Expression of Pro-inflammatory Gene IL-6 [2nd clamp visit (up to 3 months) and 3rd clamp visit (up to 6 months)]

   Adipose tissue macrophages will be isolated from subcutaneous abdominal adipose tissue, and will be quantified by fluorescence activated cell sorting (FACS) analysis. IL-6 gene expression will be examined by real-time (rt-PCR) and will provide a measure of macrophage activation at baseline, at 2nd study visit (after treatment with Vitamin D to a goal level of ≥30 ng/ml), and at 3rd study visit (goal Vitamin D level of ≥50 ng/ml). The mRNA copy number is then compared with a reference gene copy number (5 commonly used house keeping genes [HKGs]) as a ratio, which is a measure of relative gene expression.

4. Evaluated Expression of Pro-inflammatory Gene iNOS [2nd clamp visit (up to 3 months) and 3rd clamp visit (up to 6 months)]

   Adipose tissue macrophages will be isolated from subcutaneous abdominal adipose tissue, and will be quantified by fluorescence activated cell sorting (FACS) analysis. iNOS gene expression will be examined by real-time (rt-PCR) and will provide a measure of macrophage activation at baseline, at 2nd study visit (after treatment with Vitamin D to a goal level of ≥30 ng/ml), and at 3rd study visit (goal Vitamin D level of ≥50 ng/ml). The mRNA copy number is then compared with a reference gene copy number (5 commonly used house keeping genes [HKGs]) as a ratio, which is a measure of relative gene expression.

5. Evaluated Expression of Pro-inflammatory Gene PAI-1 [2nd clamp visit (up to 3 months) and 3rd clamp visit (up to 6 months)]

   Adipose tissue macrophages will be isolated from subcutaneous abdominal adipose tissue, and will be quantified by fluorescence activated cell sorting (FACS) analysis. PAI-1 gene expression will be examined by real-time (rt-PCR) and will provide a measure of macrophage activation at baseline, at 2nd study visit (after treatment with Vitamin D to a goal level of ≥30 ng/ml), and at 3rd study visit (goal Vitamin D level of ≥50 ng/ml). The mRNA copy number is then compared with a reference gene copy number (5 commonly used house keeping genes [HKGs]) as a ratio, which is a measure of relative gene expression.

Eligibility Criteria

Criteria

• Ages Eligible for Study: 21 Years to 75 Years
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: Yes

Inclusion Criteria:

• Serum 25(OH)D<20ng/ml

• Insulin Resistant based on HOMA-IR score of >3

• Able and willing to provide informed consent

• BMI 20-35

Exclusion Criteria:

• HIV/AIDS

• History of any cancer

• Sarcoidosis

• Alcohol or substance abuse

• Cushing's syndrome

• Primary hyperparathyroidism

• Nephrolithiasis

• Pregnancy or breastfeeding

• Regular visits to a tanning salon

• Hypercalcemia or hypocalcemia

• Untreated or uncontrolled hypertension

• Any chronic illness requiring medication, other than arthritis, hypertension and hyperlipidemia

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Albert Einstein College of Medicine	Bronx	New York	United States	10461

Sponsors and Collaborators

• Albert Einstein College of Medicine
• National Center for Research Resources (NCRR)

Investigators

• Principal Investigator: Meredith A Hawkins, M.D., M.S., Albert Einstein College of Medicine

Study Documents (Full-Text)

None provided.

More Information

Publications

None provided.

• Responsible Party: Meredith Hawkins, Professor of Medicine (Endocrinology), Albert Einstein College of Medicine
• ClinicalTrials.gov Identifier: NCT01354964
• Other Study ID Numbers: 2008-225; 5K23RR023335-02
• First Posted: May 17, 2011
• Last Update Posted: Nov 2, 2020
• Last Verified: Oct 1, 2020

• Individual Participant Data (IPD) Sharing Statement: No
• Plan to Share IPD: No
• Studies a U.S. FDA-regulated Drug Product: Yes
• Studies a U.S. FDA-regulated Device Product: No
• Keywords provided by Meredith Hawkins, Professor of Medicine (Endocrinology), Albert Einstein College of Medicine: Glucose Metabolism Disorders; Insulin Resistance; Endocrine System; Vitamin D
• Additional relevant MeSH terms: Insulin Resistance; Inflammation; Vitamin D; Ergocalciferols; Cholecalciferol; Vitamins

Study Results

Participant Flow

Arm/Group Title	Vitamin D	Placebo
Arm/Group Description	Participants received weekly oral vitamin D drops using a weight-based calculated dosage for up to 6 months. .	Participants received weekly oral placebo drops (similar in taste and appearance to vitamin D) for up to six months.
Period Title: Overall Study
STARTED	11	8
COMPLETED	7	7
NOT COMPLETED	4	1

Baseline Characteristics

Arm/Group Title	Vitamin D	Placebo	Total
Arm/Group Description	Participants received weekly oral vitamin D drops using a weight-based calculated dosage for up to six months. .	Participants received weekly oral placebo drops (similar in taste and appearance to vitamin D) for up to six months.	Total of all reporting groups
Overall Participants	11	8	19
Age (years) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [years]	42.0 (10.7)	47.3 (11.5)	44.2 (11.1)
Sex: Female, Male (Count of Participants)
Female	4 36.4%	2 25%	6 31.6%
Male	7 63.6%	6 75%	13 68.4%
Race/Ethnicity, Customized (Count of Participants)
White Non-Hispanic	2 18.2%	3 37.5%	5 26.3%
Black or African American	7 63.6%	5 62.5%	12 63.2%
Asian	0 0%	0 0%	0 0%
Hispanic or Latino	2 18.2%	0 0%	2 10.5%
Other	0 0%	0 0%	0 0%
Region of Enrollment (participants) [Number]
United States	11 100%	8 100%	19 100%
Hepatic Insulin Sensitivity (mg/kg/min) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [mg/kg/min]	1.28 (0.65)	1.10 (0.64)	1.20 (0.63)
Peripheral Glucose Uptake (mg/kg/min) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [mg/kg/min]	5.99 (1.18)	6.71 (2.33)	6.29 (1.74)
Evaluated expression of pro-inflammatory gene TNF-α in macrophages (Ratio of mRNA copy numbers (TNF-α/5HKGs)) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [Ratio of mRNA copy numbers (TNF-α/5HKGs)]	0.05 (0.03)	0.02 (0.02)	0.04 (0.03)
Evaluated expression of pro-inflammatory gene IL-6 in macrophages (Ratio of mRNA copy numbers (IL-6/5HKGs)) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [Ratio of mRNA copy numbers (IL-6/5HKGs)]	0.06 (0.05)	0.04 (0.05)	0.05 (0.05)
Evaluated expression of pro-inflammatory gene iNOS in macrophages (Ratio of mRNA copy numbers (iNOS/5HKGs)) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [Ratio of mRNA copy numbers (iNOS/5HKGs)]	0.007 (0.008)	0.006 (0.005)	0.007 (0.007)
Evaluated expression of pro-inflammatory gene PAI-1 in macrophages (Ratio of mRNA copy numbers (PAI-1/5HKGs)) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [Ratio of mRNA copy numbers (PAI-1/5HKGs)]	0.03 (0.03)	0.04 (0.03)	0.03 (0.03)

Outcome Measures

1. Primary Outcome

Title	Percent Change in Hepatic Insulin Sensitivity
Description	Endogenous glucose production (EGP) was assessed at each study visit to evaluate hepatic insulin sensitivity. Percent change between the EGP at baseline and second visit (after treatment for up to 3 months with Vitamin D to reach a target level of ≥30 ng/ml), and baseline and third visits (after treatment for up to 6 months with Vitamin D in order to reach a target level of ≥50 ng/ml) will be calculated.
Time Frame	2nd clamp visit (after up to 3 months) and 3rd clamp visit (after up to 6 months)

Outcome Measure Data

Analysis Population Description

Hepatic insulin sensitivity was measured in 7 (out of 11) participants in the Vitamin D group and in 7 (out of 8) participants in the placebo group at the 3rd study visit due to loss to follow up, withdrawal, or difficulties in obtaining IV access.

Arm/Group Title	Vitamin D	Placebo
Arm/Group Description	Participants received weekly oral vitamin D drops using a weight-based calculated dosage to reach a target level of ≥30 ng/ml at second visit and a goal level of ≥50 ng/ml at third visit.	Participants received weekly oral placebo drops (similar in taste and appearance to vitamin D).
Measure Participants	11	8
Percent change between baseline and 2nd visit	-19.2 (11.6)	5.8 (22.5)
Percent change between baseline and 3rd visit	-33.66 (7.5)	113.7 (65.5)

2. Secondary Outcome

Title	Percent Change in Peripheral Glucose Uptake
Description	The rate of glucose uptake to determine peripheral insulin sensitivity was measured using the rate of disappearance (Rd) of glucose at each study visit. Percent change between the Rd at baseline and second visit (after treatment with Vitamin D for up to 3 months to target level of ≥30 ng/ml), and baseline and third visits (after treatment with Vitamin D for up to 6 months to target level of ≥50 ng/ml) will be calculated.
Time Frame	2nd clamp visit (up to 3 months) and 3rd clamp visit (up to 6 months)

Outcome Measure Data

Analysis Population Description

Hepatic insulin sensitivity was measured in 7 (out of 11) participants in the Vitamin D group and in 6 (out of 8) participants in the placebo group at the 3rd study visit due to loss to follow up, withdrawal, or difficulties in obtaining IV access.

Arm/Group Title	Vitamin D	Placebo
Arm/Group Description	Participants received weekly oral vitamin D drops using a weight-based calculated dosage to reach a target level of ≥30 ng/ml at second visit and a goal level of ≥50 ng/ml at third visit.	Participants received weekly oral placebo drops (similar in taste and appearance to vitamin D).
Measure Participants	11	8
Percent change between baseline and 2nd visit	0.48 (8.1)	-2.54 (5.8)
Percent change between baseline and 3rd visit	-0.98 (7.2)	1.75 (10.1)

3. Secondary Outcome

Title	Evaluated Expression of Pro-inflammatory Gene TNF-α
Description	Adipose tissue macrophages will be isolated from subcutaneous abdominal adipose tissue, and will be quantified by fluorescence activated cell sorting (FACS) analysis. TNF-α gene expression will be examined by real-time (rt-PCR) and will provide a measure of macrophage activation at baseline, at 2nd study visit (after treatment with Vitamin D to a goal level of ≥30 ng/ml), and at 3rd study visit (goal Vitamin D level of ≥50 ng/ml). The mRNA copy number is then compared with a reference gene copy number (5 commonly used house keeping genes [HKGs]) as a ratio, which is a measure of relative gene expression.
Time Frame	2nd clamp visit (up to 3 months) and 3rd clamp visit (up to 6 months)

Outcome Measure Data

Analysis Population Description

TNF-α expression was measured in 7 (out of 11) participants in the Vitamin D group and in 4 (out of 8) participants in the placebo group at the 3rd study visit due to loss to follow up, withdrawal, or difficulties in obtaining IV access. Additionally, 2 participants in placebo group did not provide fat biopsy samples for the study.

Arm/Group Title	Vitamin D	Placebo
Arm/Group Description	Participants received weekly oral vitamin D drops using a weight-based calculated dosage to reach a target level of ≥30 ng/ml at second visit and a goal level of ≥50 ng/ml at third visit.	Participants in this group were assigned to receive weekly oral placebo drops (similar in taste and appearance to vitamin D) for three months.
Measure Participants	11	6
TNF-α (2nd study visit)	0.023 (0.006)	0.021 (0.007)
TNF-α (3rd study visit)	0.036 (0.009)	0.008 (0.003)

4. Secondary Outcome

Title	Evaluated Expression of Pro-inflammatory Gene IL-6
Description	Adipose tissue macrophages will be isolated from subcutaneous abdominal adipose tissue, and will be quantified by fluorescence activated cell sorting (FACS) analysis. IL-6 gene expression will be examined by real-time (rt-PCR) and will provide a measure of macrophage activation at baseline, at 2nd study visit (after treatment with Vitamin D to a goal level of ≥30 ng/ml), and at 3rd study visit (goal Vitamin D level of ≥50 ng/ml). The mRNA copy number is then compared with a reference gene copy number (5 commonly used house keeping genes [HKGs]) as a ratio, which is a measure of relative gene expression.
Time Frame	2nd clamp visit (up to 3 months) and 3rd clamp visit (up to 6 months)

Outcome Measure Data

Analysis Population Description

IL-6 expression was measured in 7 (out of 11) participants in the Vitamin D group. For the placebo group, 6 of the 8 participants provided fat biopsy samples for the 2nd visit, and 4 participants provided for the 3rd visit due to loss to follow up, withdrawal, or difficulties in obtaining IV access.

Arm/Group Title	Vitamin D	Placebo
Arm/Group Description	Participants in this group were assigned to receive weekly oral vitamin D drops using a weight-based calculated dosage for three months. .	Participants in this group were assigned to receive weekly oral placebo drops (similar in taste and appearance to vitamin D) for three months.
Measure Participants	11	6
IL-6 (2nd study visit)	0.019 (0.006)	0.047 (0.018)
IL-6 (3rd study visit)	0.022 (0.007)	0.050 (0.029)

5. Secondary Outcome

Title	Evaluated Expression of Pro-inflammatory Gene iNOS
Description	Adipose tissue macrophages will be isolated from subcutaneous abdominal adipose tissue, and will be quantified by fluorescence activated cell sorting (FACS) analysis. iNOS gene expression will be examined by real-time (rt-PCR) and will provide a measure of macrophage activation at baseline, at 2nd study visit (after treatment with Vitamin D to a goal level of ≥30 ng/ml), and at 3rd study visit (goal Vitamin D level of ≥50 ng/ml). The mRNA copy number is then compared with a reference gene copy number (5 commonly used house keeping genes [HKGs]) as a ratio, which is a measure of relative gene expression.
Time Frame	2nd clamp visit (up to 3 months) and 3rd clamp visit (up to 6 months)

Outcome Measure Data

Analysis Population Description

iNOS expression was measured in 7 (out of 11) participants in the Vitamin D group. For the placebo group, 6 of the 8 participants provided fat biopsy samples for the 2nd visit, and 4 participants provided for the 3rd visit due to loss to follow up, withdrawal, or difficulties in obtaining IV access.

Arm/Group Title	Vitamin D	Placebo
Arm/Group Description	Participants in this group were assigned to receive weekly oral vitamin D drops using a weight-based calculated dosage for three months. .	Participants in this group were assigned to receive weekly oral placebo drops (similar in taste and appearance to vitamin D) for three months.
Measure Participants	11	6
iNOS (2nd study visit)	0.004 (0.002)	0.008 (0.005)
iNOS (3rd study visit)	0.003 (0.001)	0.005 (0.002)

6. Secondary Outcome

Title	Evaluated Expression of Pro-inflammatory Gene PAI-1
Description	Adipose tissue macrophages will be isolated from subcutaneous abdominal adipose tissue, and will be quantified by fluorescence activated cell sorting (FACS) analysis. PAI-1 gene expression will be examined by real-time (rt-PCR) and will provide a measure of macrophage activation at baseline, at 2nd study visit (after treatment with Vitamin D to a goal level of ≥30 ng/ml), and at 3rd study visit (goal Vitamin D level of ≥50 ng/ml). The mRNA copy number is then compared with a reference gene copy number (5 commonly used house keeping genes [HKGs]) as a ratio, which is a measure of relative gene expression.
Time Frame	2nd clamp visit (up to 3 months) and 3rd clamp visit (up to 6 months)

Outcome Measure Data

Analysis Population Description

PAI-1 expression was measured in 7 (out of 11) participants in the Vitamin D group. For the placebo group, 6 of the 8 participants provided fat biopsy samples for the 2nd visit, and 4 participants provided for the 3rd visit due to loss to follow up, withdrawal, or difficulties in obtaining IV access.

Arm/Group Title	Vitamin D	Placebo
Arm/Group Description	Participants in this group were assigned to receive weekly oral vitamin D drops using a weight-based calculated dosage for three months. .	Participants in this group were assigned to receive weekly oral placebo drops (similar in taste and appearance to vitamin D) for three months.
Measure Participants	11	6
PAI-1 (2nd study visit)	0.020 (0.008)	0.032 (0.012)
PAI-1 (3rd study visit)	0.008 (0.005)	0.019 (0.008)

Adverse Events

Time Frame	Time enrolled in study (approximately 6 months)
Arm/Group Title	Vitamin D		Placebo
Arm/Group Description	Participants in this group were assigned to receive weekly oral vitamin D drops using a weight-based calculated dosage for six months. .		Participants in this group were assigned to receive weekly oral placebo drops (similar in taste and appearance to vitamin D) for six months.
All Cause Mortality
	Vitamin D		Placebo
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/11 (0%)		0/8 (0%)
Serious Adverse Events
	Vitamin D		Placebo
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	1/11 (9.1%)		0/8 (0%)
Ear and labyrinth disorders
benign paroxysmal vertigo	1/11 (9.1%)	1	0/8 (0%)	0
Other (Not Including Serious) Adverse Events
	Vitamin D		Placebo
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	1/11 (9.1%)		0/8 (0%)
Nervous system disorders
pre-syncopal episode	1/11 (9.1%)	1	0/8 (0%)	0

Limitations/Caveats

[Not Specified]

More Information

Certain Agreements

All Principal Investigators ARE employed by the organization sponsoring the study.

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.