Effect of Vitamin D Repletion on Insulin Resistance and Systemic Inflammation

• ClinicalTrials.gov Identifier: NCT01354964
• Recruitment Status: Completed
• First Posted: May 17, 2011
• Results First Posted: March 9, 2020
• Last Update Posted: November 2, 2020
• Sponsor: Albert Einstein College of Medicine
• Collaborator: National Center for Research Resources (NCRR)
• Information provided by (Responsible Party): Meredith Hawkins, Albert Einstein College of Medicine

• Study Type: Interventional
• Study Design: Allocation: Randomized; Intervention Model: Parallel Assignment; Masking: Double (Participant, Investigator); Primary Purpose: Treatment
• Condition: Insulin Resistance
• Interventions: Drug: Vitamin D; Drug: Placebo
• Enrollment: 19

Participant Flow

Recruitment Details
Pre-assignment Details

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
Reason Not Completed
Lost to Follow-up	2	0
IV access issues	1	1
Physician Decision	1	0

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 Number of Baseline Participants		11	8	19
Baseline Analysis Population Description		[Not Specified]
Age, Continuous; Mean (Standard Deviation); Unit of measure: Years
	Number Analyzed	11 participants	8 participants	19 participants
		42.0 (10.7)	47.3 (11.5)	44.2 (11.1)
Sex: Female, Male; Measure Type: Count of Participants; Unit of measure: Participants
	Number Analyzed	11 participants	8 participants	19 participants
	Female	4 36.4%	2 25.0%	6 31.6%
	Male	7 63.6%	6 75.0%	13 68.4%
Race/Ethnicity, Customized; Measure Type: Count of Participants; Unit of measure: Participants
White Non-Hispanic	Number Analyzed	11 participants	8 participants	19 participants
		2 18.2%	3 37.5%	5 26.3%
Black or African American	Number Analyzed	11 participants	8 participants	19 participants
		7 63.6%	5 62.5%	12 63.2%
Asian	Number Analyzed	11 participants	8 participants	19 participants
		0 0.0%	0 0.0%	0 0.0%
Hispanic or Latino	Number Analyzed	11 participants	8 participants	19 participants
		2 18.2%	0 0.0%	2 10.5%
Other	Number Analyzed	11 participants	8 participants	19 participants
		0 0.0%	0 0.0%	0 0.0%
Region of Enrollment; Measure Type: Number; Unit of measure: Participants
United States	Number Analyzed	11 participants	8 participants	19 participants
		11	8	19
Hepatic Insulin Sensitivity [1]; Mean (Standard Deviation); Unit of measure: Mg/kg/min
	Number Analyzed	11 participants	8 participants	19 participants
		1.28 (0.65)	1.10 (0.64)	1.20 (0.63)
		[1] Measure Description: We examine the endogenous glucose production (EGP) to assess hepatic insulin sensitivity at baseline.
Peripheral Glucose Uptake [1]; Mean (Standard Deviation); Unit of measure: Mg/kg/min
	Number Analyzed	11 participants	8 participants	19 participants
		5.99 (1.18)	6.71 (2.33)	6.29 (1.74)
		[1] Measure Description: We examine the rate of glucose uptake to determine peripheral insulin sensitivity using rate of disappearance (Rd) of glucose.
Evaluated expression of pro-inflammatory gene TNF-α in macrophages [1] [2]; Mean (Standard Deviation); Unit of measure: Ratio of mRNA copy numbers (TNF-α/5HKGs)
	Number Analyzed	11 participants	6 participants	17 participants
		0.05 (0.03)	0.02 (0.02)	0.04 (0.03)
		[1] Measure 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 polymerase chain reaction (rt-PCR) and will provide a measure of macrophage activation. 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.; [2] Measure Analysis Population Description: TNFα was measured in 6 participants in placebo group.
Evaluated expression of pro-inflammatory gene IL-6 in macrophages [1] [2]; Mean (Standard Deviation); Unit of measure: Ratio of mRNA copy numbers (IL-6/5HKGs)
	Number Analyzed	11 participants	6 participants	17 participants
		0.06 (0.05)	0.04 (0.05)	0.05 (0.05)
		[1] Measure 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 polymerase chain reaction (rt-PCR) and will provide a measure of macrophage activation. 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.; [2] Measure Analysis Population Description: IL-6 was measured in 6 participants in placebo group.
Evaluated expression of pro-inflammatory gene iNOS in macrophages [1] [2]; Mean (Standard Deviation); Unit of measure: Ratio of mRNA copy numbers (iNOS/5HKGs)
	Number Analyzed	11 participants	6 participants	17 participants
		0.007 (0.008)	0.006 (0.005)	0.007 (0.007)
		[1] Measure 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 polymerase chain reaction (rt-PCR) and will provide a measure of macrophage activation. 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.; [2] Measure Analysis Population Description: iNOS was measured in 6 participants in placebo group.
Evaluated expression of pro-inflammatory gene PAI-1 in macrophages [1] [2]; Mean (Standard Deviation); Unit of measure: Ratio of mRNA copy numbers (PAI-1/5HKGs)
	Number Analyzed	11 participants	6 participants	17 participants
		0.03 (0.03)	0.04 (0.03)	0.03 (0.03)
		[1] Measure 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 polymerase chain reaction (rt-PCR) and will provide a measure of macrophage activation. 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.; [2] Measure Analysis Population Description: PAI-1 was measured in 6 participants in placebo group.

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).
Overall Number of Participants Analyzed		11	8
Mean (Standard Error); Unit of Measure: percent change
Percent change between baseline and 2nd visit	Number Analyzed	11 participants	8 participants
		-19.2 (11.6)	5.8 (22.5)
Percent change between baseline and 3rd visit	Number Analyzed	7 participants	7 participants
		-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).
Overall Number of Participants Analyzed		11	8
Mean (Standard Error); Unit of Measure: percent change
Percent change between baseline and 2nd visit	Number Analyzed	11 participants	8 participants
		0.48 (8.1)	-2.54 (5.8)
Percent change between baseline and 3rd visit	Number Analyzed	7 participants	6 participants
		-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.
Overall Number of Participants Analyzed		11	6
Mean (Standard Error); Unit of Measure: Ratio of mRNA copy numbers (TNF-α/5HKGs)
TNF-α (2nd study visit)	Number Analyzed	11 participants	6 participants
		0.023 (0.006)	0.021 (0.007)
TNF-α (3rd study visit)	Number Analyzed	7 participants	4 participants
		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.
Overall Number of Participants Analyzed		11	6
Mean (Standard Error); Unit of Measure: Ratio of mRNA copy numbers (IL-6/5HKGs)
IL-6 (2nd study visit)	Number Analyzed	11 participants	6 participants
		0.019 (0.006)	0.047 (0.018)
IL-6 (3rd study visit)	Number Analyzed	7 participants	4 participants
		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.
Overall Number of Participants Analyzed		11	6
Mean (Standard Error); Unit of Measure: Ratio of mRNA copy numbers (iNOS/5HKGs)
iNOS (2nd study visit)	Number Analyzed	11 participants	6 participants
		0.004 (0.002)	0.008 (0.005)
iNOS (3rd study visit)	Number Analyzed	7 participants	4 participants
		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.
Overall Number of Participants Analyzed		11	6
Mean (Standard Error); Unit of Measure: Ratio of mRNA copy numbers (PAI-1/5HKGs)
PAI-1 (2nd study visit)	Number Analyzed	11 participants	6 participants
		0.020 (0.008)	0.032 (0.012)
PAI-1 (3rd study visit)	Number Analyzed	7 participants	4 participants
		0.008 (0.005)	0.019 (0.008)

Adverse Events

Time Frame	Time enrolled in study (approximately 6 months)
Adverse Event Reporting Description	[Not Specified]
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 (%)		Affected / at Risk (%)
Total	0/11 (0.00%)		0/8 (0.00%)
Serious Adverse Events
	Vitamin D		Placebo
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	1/11 (9.09%)		0/8 (0.00%)
Ear and labyrinth disorders
benign paroxysmal vertigo *	1/11 (9.09%)	1	0/8 (0.00%)	0
* Indicates events were collected by non-systematic assessment
Other (Not Including Serious) Adverse Events
Frequency Threshold for Reporting Other Adverse Events	5%
	Vitamin D		Placebo
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	1/11 (9.09%)		0/8 (0.00%)
Nervous system disorders
pre-syncopal episode *	1/11 (9.09%)	1	0/8 (0.00%)	0
* Indicates events were collected by non-systematic assessment

Limitations and Caveats

[Not Specified]

More Information

Certain Agreements

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

Results Point of Contact

• Name/Title: Dr. Meredith Hawkins
• Organization: Albert Einstein College of Medicine
• Phone: 7184302903 ext 2903
• EMail: meredith.hawkins@einsteinmed.org

• Responsible Party: Meredith Hawkins, Albert Einstein College of Medicine
• ClinicalTrials.gov Identifier: NCT01354964
• Other Study ID Numbers: 2008-225; 5K23RR023335-02 ( U.S. NIH Grant/Contract )
• First Submitted: May 12, 2011
• First Posted: May 17, 2011
• Results First Submitted: April 12, 2019
• Results First Posted: March 9, 2020
• Last Update Posted: November 2, 2020