Bioavailability of SPMs in Obese Humans

• Sponsor: University of North Carolina, Chapel Hill (Other)
• Overall Status: Completed
• CT.gov ID: NCT04701138
• Collaborator: Metagenics, Inc. (Industry), North Carolina Translational and Clinical Sciences Institute (Other)
• Enrollment: 24
• Location: 1
• Arm: 1
• Actual Duration (Months): 4.4
• Patients Per Site Per Month: 5.5

Study Details

Study Description

Brief Summary

Research Question: Does 4 weeks of supplementation with 'SPM Active' lead to a statistically significant increase in plasma SPM concentration for obese human subjects? Primary Aim 1: To compare plasma SPM concentrations and immunological fitness pre- and post- oral SPM administration in the obese.

• Aim 1a: To quantify plasma SPM concentrations in plasma (pg/mL), serum (pg/mL) and PBMCs before and after 4 weeks of supplementation with 'SPM Active.' The concentration of SPMs in plasma, in addition to other PUFA-derived metabolites that share the same enzymatic pathways as SPMs, will be established at baseline and post-intervention using mass spectrometry-based metabololipidomics.

• Aim 1b: To measure in vitro antibody responses of B cells in PBMC pool with in vitro stimulation and cytokine production before and after 4 weeks of supplementation with 'SPM Active.' In addition, researchers will quantify the relative abundance of differing immune cell populations.

Condition or Disease	Intervention/Treatment	Phase
Obesity Adult Onset	Dietary Supplement: SPM Active	N/A

Detailed Description

Purpose: Specialized pro-resolving mediators (SPMs) are a superfamily of lipid metabolites, predominantly derived from the n-3 polyunsaturated fatty acids (PUFAs) eicosapentaenoic (EPA) and docosahexaenoic acids (DHA). Previous research has established that obese mice and humans have lower circulating levels of SPMs relative to lean controls. In this study, SPMs will be administered as a dietary supplement to obese human subjects to establish: 1) their bioavailability in plasma, serum and peripheral blood mononuclear cells (PBMCs) and 2) their effects on immune cell abundance and in vitro antibody production. The rationale for focusing on immune cells is that SPMs may be targeting their abundance and phenotype. This study does not intend to make any health or health-related claims.

Participants: A total of 24 (n=12 men + 12 women) obese (BMI 30-40 kg/m2) euglycemic and pre-diabetic subjects (fasting glucose 70-125 mg/dL or HbA1c of 5.7-6.4%) aged 50-65 years will be recruited by Dr. Erik Butler from the UNC Family Medicine Center in Chapel Hill.

Procedures (methods): This is a non-randomized uncontrolled clinical trial. The study will provide the intervention 'SPM Active' provided by Metagenics. All subjects will be advised to take 4 capsules per day (2 capsules with breakfast and 2 capsules with dinner) of 'SPM Active' for 4 weeks total. Each capsule contains 145 mg of SPMs for a total daily dose of 580 mg. Fasting blood will be drawn pre- and post-intervention using phlebotomy available under the direction of Dr. Butler. The scientific approach will rely on mass-spectrometry based metabololipidomics, immunophenotyping with flow cytometry, and anthropomorphic/blood pressure/BMI measurements (anthropometric measures are only intended for use in statistical analysis for confounding variables).

Study Design

• Study Type: Interventional
• Actual Enrollment: 24 participants
• Allocation: N/A
• Intervention Model: Single Group Assignment
• Masking: None (Open Label)
• Primary Purpose: Basic Science
• Official Title: Bioavailability of Specialized Pro-resolving Mediators in Obese Humans
• Actual Study Start Date: Feb 4, 2021
• Actual Primary Completion Date: Jun 2, 2021
• Actual Study Completion Date: Jun 18, 2021

Arms and Interventions

Arm	Intervention/Treatment
Experimental: Dietary Supplement All subjects will receive the intervention (SPM Active Supplement)	Dietary Supplement: SPM Active; Each capsule contains 145 mg of SPMs (fractionated marine lipids standardized to 18-HEPE, 14-HDHA, and 17-HDHA). All subjects will take 4 capsules orally each day for a total daily dose of 580 mg.

Outcome Measures

Primary Outcome Measures

1. Mean Pro-inflammatory & Pro-resolving Metabolites [From Baseline (Week 1/Day1) through 28 to 30 days of supplementation]

   Mass spectrometry metabololipidomics analysis will be performed on plasma samples from pre/post supplementation blood draws to measure SPM concentrations. The study was powered to measure the following molecules of interest: 14-HDHA, 17-HDHA, and 18-HEPE.

Secondary Outcome Measures

1. Mean White Blood Cell Populations [From Baseline (Week 1/Day1) through 28 to 30 days of supplementation]

   Immunological phenotyping of blood peripheral mononuclear cells (PBMC) using flow cytometry will identify key immune cell populations pre/post supplementation. PBMC analyses represent B cell populations, monocyte populations, natural killer cell populations, and T cell populations. The relative abundance was calculated using two different flow cytometry panels with fluorescently labeled antibodies. The first panel measured the relative abundance of all B cell subsets, monocyte subsets, and NK cell subsets (i.e., all subsets within error add up to 1.0). The second panel measured the relative abundance of CD4 T cell subsets, CD8 T cell subsets, and NKT cells (i.e., all subsets within error add up to 1.0).

Other Outcome Measures

1. Antibody Concentrations in Culture [From Baseline (Week 1/Day1) through 28 to 30 days of supplementation]

   B cells isolated from collected blood samples will be cultured in-vitro and stimulated with an antigen & produced antibody concentrations will be measured via ELISA. This will be done for pre & post blood samples.

Eligibility Criteria

Criteria

• Ages Eligible for Study: 50 Years to 65 Years
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: No

Inclusion Criteria:

• 24 (n=12 male + 12 female) obese (BMI 30-40 kg/m^2) subjects with age 50-65 years who are euglycemic and pre-diabetic (i.e. fasting glucose 70-125 mg/dL or HbA1c of 5.7-6.4%).

• Only post-menopausal females will be recruited in the female cohort to reduce the confounding effects of estrogen on lipid metabolism during supplementation.

Exclusion Criteria:

• Those with fasting glucose values > 126 mg/dL or known type 2 diabetes

• Females who are pre-menopausal, pregnant, planning to become pregnant, breastfeeding or lactating

• Subjects consuming n-3 PUFA supplements in the last 3 months prior to enrollment, high consumption of fatty fish (>2 servings per week), and subjects with active autoimmune disease, liver disease, coagulopathy, hypothyroidism, known allergy to fish or shellfish, inability to give informed consent, or taking anticoagulants (e.g. warfarin and direct-acting anticoagulants), those taking estrogen or testosterone, and anyone taking daily aspirin, NSAIDs, or active asthma medications.

• Subjects receiving immunomodulatory or immunosuppressant therapy (corticosteroids or monoclonal antibodies) in the 4 weeks prior to study enrollment, and subjects with known active malignancy or undergoing treatment for malignancy will be excluded.

• Subjects who test positive for COVID-19 or have tested positive in the past will be excluded. Those who report COVID-19 or flu-like symptoms will be excluded, as well as those that fail to pass COVID-19 screening at baseline.

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	UNC Chapel Hill Family Medicine Center	Chapel Hill	North Carolina	United States	27514

Sponsors and Collaborators

• University of North Carolina, Chapel Hill
• Metagenics, Inc.
• North Carolina Translational and Clinical Sciences Institute

Investigators

• Principal Investigator: Saame R Shaikh, PhD, University of North Carolina, Chapel Hill
• Study Director: Erik Butler, DO, UNC Family Medicine

Study Documents (Full-Text)

• Study Protocol and Statistical Analysis Plan - Jun 5, 2020
• Informed Consent Form - Jan 29, 2021

More Information

Publications

• Crouch MJ, Kosaraju R, Guesdon W, Armstrong M, Reisdorph N, Jain R, Fenton J, Shaikh SR. Frontline Science: A reduction in DHA-derived mediators in male obesity contributes toward defects in select B cell subsets and circulating antibody. J Leukoc Biol. 2019 Aug;106(2):241-257. doi: 10.1002/JLB.3HI1017-405RR. Epub 2018 Dec 21.
• Kosaraju R, Guesdon W, Crouch MJ, Teague HL, Sullivan EM, Karlsson EA, Schultz-Cherry S, Gowdy K, Bridges LC, Reese LR, Neufer PD, Armstrong M, Reisdorph N, Milner JJ, Beck M, Shaikh SR. B Cell Activity Is Impaired in Human and Mouse Obesity and Is Responsive to an Essential Fatty Acid upon Murine Influenza Infection. J Immunol. 2017 Jun 15;198(12):4738-4752. doi: 10.4049/jimmunol.1601031. Epub 2017 May 12.
• López-Vicario C, Titos E, Walker ME, Alcaraz-Quiles J, Casulleras M, Durán-Güell M, Flores-Costa R, Pérez-Romero N, Forné M, Dalli J, Clària J. Leukocytes from obese individuals exhibit an impaired SPM signature. FASEB J. 2019 Jun;33(6):7072-7083. doi: 10.1096/fj.201802587R. Epub 2019 Mar 6.
• Serhan CN, Levy BD. Resolvins in inflammation: emergence of the pro-resolving superfamily of mediators. J Clin Invest. 2018 Jul 2;128(7):2657-2669. doi: 10.1172/JCI97943. Epub 2018 May 14. Review.

• Responsible Party: University of North Carolina, Chapel Hill
• ClinicalTrials.gov Identifier: NCT04701138
• Other Study ID Numbers: 20-0131; 550KR242033
• First Posted: Jan 8, 2021
• Last Update Posted: Jun 15, 2022
• Last Verified: Mar 1, 2022

• Individual Participant Data (IPD) Sharing Statement: Yes
• Plan to Share IPD: Yes
• Studies a U.S. FDA-regulated Drug Product: No
• Studies a U.S. FDA-regulated Device Product: No

Study Results

Participant Flow

Arm/Group Title	Dietary Supplement
Arm/Group Description	All subjects will receive the intervention (SPM Active Supplement) SPM Active: Each capsule contains 145 mg of SPMs (fractionated marine lipids standardized to 18-HEPE, 14-HDHA, and 17-HDHA). All subjects will take 4 capsules orally each day for a total daily dose of 580 mg.
Period Title: Overall Study
STARTED	24
COMPLETED	23
NOT COMPLETED	1

Baseline Characteristics

Arm/Group Title	Dietary Supplement
Arm/Group Description	All subjects will receive the intervention (SPM Active Supplement) SPM Active: Each capsule contains 145 mg of SPMs (fractionated marine lipids standardized to 18-HEPE, 14-HDHA, and 17-HDHA). All subjects will take 4 capsules orally each day for a total daily dose of 580 mg.
Overall Participants	23
Age (years) [Mean (Full Range) ]
Mean (Full Range) [years]	56
Sex: Female, Male (Count of Participants)
Female	13 56.5%
Male	10 43.5%
Ethnicity (NIH/OMB) (Count of Participants)
Hispanic or Latino	0 0%
Not Hispanic or Latino	23 100%
Unknown or Not Reported	0 0%
Race (NIH/OMB) (Count of Participants)
American Indian or Alaska Native	0 0%
Asian	1 4.3%
Native Hawaiian or Other Pacific Islander	0 0%
Black or African American	5 21.7%
White	16 69.6%
More than one race	0 0%
Unknown or Not Reported	1 4.3%
Region of Enrollment (Count of Participants)
United States	23 100%
Weight (Kg) [Median (Full Range) ]
Median (Full Range) [Kg]	96.4
Height (cm) [Median (Full Range) ]
Median (Full Range) [cm]	170.2
Body Mass Index (BMI) (Kg/m^2) [Median (Full Range) ]
Median (Full Range) [Kg/m^2]	33.1

Outcome Measures

1. Primary Outcome

Title	Mean Pro-inflammatory & Pro-resolving Metabolites
Description	Mass spectrometry metabololipidomics analysis will be performed on plasma samples from pre/post supplementation blood draws to measure SPM concentrations. The study was powered to measure the following molecules of interest: 14-HDHA, 17-HDHA, and 18-HEPE.
Time Frame	From Baseline (Week 1/Day1) through 28 to 30 days of supplementation

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Dietary Supplement
Arm/Group Description	All subjects will receive the intervention (SPM Active Supplement) SPM Active: Each capsule contains 145 mg of SPMs (fractionated marine lipids standardized to 18-HEPE, 14-HDHA, and 17-HDHA). All subjects will take 4 capsules orally each day for a total daily dose of 580 mg.
Measure Participants	23
Pre 14-HDHA	15.6 (11.5)
Post 14-HDHA	12.2 (5.4)
Pre 17-HDHA	6.6 (4.9)
Post 17-HDHA	8.1 (4.6)
Pre 18-HEPE	8.0 (6.2)
Post 18-HEPE	11.3 (10.3)

2. Secondary Outcome

Title	Mean White Blood Cell Populations
Description	Immunological phenotyping of blood peripheral mononuclear cells (PBMC) using flow cytometry will identify key immune cell populations pre/post supplementation. PBMC analyses represent B cell populations, monocyte populations, natural killer cell populations, and T cell populations. The relative abundance was calculated using two different flow cytometry panels with fluorescently labeled antibodies. The first panel measured the relative abundance of all B cell subsets, monocyte subsets, and NK cell subsets (i.e., all subsets within error add up to 1.0). The second panel measured the relative abundance of CD4 T cell subsets, CD8 T cell subsets, and NKT cells (i.e., all subsets within error add up to 1.0).
Time Frame	From Baseline (Week 1/Day1) through 28 to 30 days of supplementation

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Dietary Supplement
Arm/Group Description	All subjects will receive the intervention (SPM Active Supplement) SPM Active: Each capsule contains 145 mg of SPMs (fractionated marine lipids standardized to 18-HEPE, 14-HDHA, and 17-HDHA). All subjects will take 4 capsules orally each day for a total daily dose of 580 mg.
Measure Participants	23
Pre CD4+ T Cells	0.32 (0.07)
Post CD4+ T Cells	0.33 (0.05)
Pre Activated CD4+ T Cells	0.02 (0.01)
Post Activated CD4+ T Cells	0.03 (0.01)
Pre Non-Activated CD4+ T Cells	0.30 (0.07)
Post Non-Activated CD4+ T Cells	0.30 (0.05)
Pre CD8 T Cells	0.17 (0.07)
Post CD8 T Cells	0.16 (0.05)
Pre Activated CD8 T Cells	0.02 (0.01)
Post Activated CD8 T Cells	0.03 (0.02)
Pre Non-Activated CD8 T Cells	0.15 (0.06)
Post Non-Activated CD8 T Cells	0.13 (0.03)
Pre NKT Cells	0.02 (0.01)
Post NKT Cells	0.02 (0.01)
Pre B Cells	0.39 (0.08)
Post B Cells	0.37 (0.11)
Pre Plasma Cells	0.02 (0.01)
Post Plasma Cells	0.02 (0.01)
Pre Follicular B Cells	0.01 (0.01)
Post Follicular B Cells	0.01 (0.00)
Pre B Regs	0.08 (0.04)
Post B Regs	0.07 (0.04)
Pre Classical Monocytes	0.06 (0.02)
Post Classical Monocytes	0.06 (0.03)
Pre Intermediate Monocytes	0.00 (0.00)
Post Intermediate Monocytes	0.01 (0.00)
Pre Non-Classical Monocytes	0.11 (0.06)
Post Non-Classical Monocytes	0.10 (0.07)
Pre NK CD3-CD56HICD16-	0.01 (0.00)
Post NK CD3-CD56HICD16-	0.02 (0.03)
Pre NK CD3-CD56HICD16+	0.01 (0.01)
Post NK CD3-CD56HICD16+	0.02 (0.01)
Pre NK CD3-CD56dimCD16-	0.23 (0.09)
Post NK CD3-CD56dimCD16-	0.22 (0.08)
Pre NK CD3-CD56dimCD16+	0.02 (0.01)
Post NK CD3-CD56dimCD16+	0.02 (0.01)
Pre NK CD3-CD56-CD16+	0.05 (0.03)
Post NK CD3-CD56-CD16+	0.08 (0.08)

3. Other Pre-specified Outcome

Title	Antibody Concentrations in Culture
Description	B cells isolated from collected blood samples will be cultured in-vitro and stimulated with an antigen & produced antibody concentrations will be measured via ELISA. This will be done for pre & post blood samples.
Time Frame	From Baseline (Week 1/Day1) through 28 to 30 days of supplementation

Outcome Measure Data

Analysis Population Description

One participant's sample was removed from all B cell isolation and stimulation studies (ELISA data) due to PBMC cell death upon sample freezing.

Arm/Group Title	Dietary Supplement
Arm/Group Description	All subjects will receive the intervention (SPM Active Supplement) SPM Active: Each capsule contains 145 mg of SPMs (fractionated marine lipids standardized to 18-HEPE, 14-HDHA, and 17-HDHA). All subjects will take 4 capsules orally each day for a total daily dose of 580 mg.
Measure Participants	22
Pre IgM	465801.89 (217732.43)
Post IgM	457833.05 (238874.79)
Pre IgG	1024891.3 (394612.17)
Post IgG	789525.69 (320595.08)

Adverse Events

Time Frame	From the time of signing informed consent through the last blood draw, ranging from 28 to 30 days.
Arm/Group Title	Dietary Supplement
Arm/Group Description	All subjects will receive the intervention (SPM Active Supplement) SPM Active: Each capsule contains 145 mg of SPMs (fractionated marine lipids standardized to 18-HEPE, 14-HDHA, and 17-HDHA). All subjects will take 4 capsules orally each day for a total daily dose of 580 mg.
All Cause Mortality
	Dietary Supplement
	Affected / at Risk (%)	# Events
Total	0/24 (0%)
Serious Adverse Events
	Dietary Supplement
	Affected / at Risk (%)	# Events
Total	0/24 (0%)
Other (Not Including Serious) Adverse Events
	Dietary Supplement
	Affected / at Risk (%)	# Events
Total	0/24 (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.