Progress 02/15/15 to 02/14/20
Outputs
Target Audience: Researchers in the fields of inflammation and lipid metabolism; registered dieticians; physicians individuals with metabolic syndrome, elevated chronic inflammation, elevated plasma lipid (triglyceride and cholesterol) levels Changes/Problems:This study has a randomized, placebo-controlled, double-blind, crossover design, and was designed with a placebo lead-in phase lasting 4 weeks followed by randomization to either EPA or DHA for 12 weeks, then by a placebo phase of 12 weeks, and finally by a crossover to either DHA or EPA for 12 weeks, for a total length of the study of 40 weeks. Before the start of the study, to increase study subjects' recruitment, retention and compliance, we have reduced the 12-week EPA, DHA and placebo phases to 10 weeks each, resulting in a total length of the study of 34 weeks. This change has not affected the study design or subjects' safety, and still allowed us to reliably measure the inflammatory and plasma lipid outcomes as originally proposed in the grant, while enhancing feasibility and retention. In addition, we have changed one of the inclusion criteria of the study, namely plasma triglyceride levels, from 150-500 mg/dL to 90- 500 mg/dL. This change has increased the number of subjects eligible to enroll into the study. These changes to the original protocol have been approved by our Institutional Review Board (IRB) What opportunities for training and professional development has the project provided?The project has provided the opportunity to train a doctoral student in methodologies and data analysis. Specifically, the graduate student has been trained in the isolation of peripheral blood mononuclear cells (PBMC) and the further isolation of monocytes by selective immunoseparation. The student also trained in the culture and stimulation of monocytes with lipopolysaccharide, the isolation of monocyte RNA, and the measurement of target messenger RNAs by real time PCR. In addition, the doctoral student has trained in complex bioinformatic analysis of lipidomics and RNA-sequencing data. How have the results been disseminated to communities of interest?The results of the study have been presented at several meetings and disseminated through published abstracts. Two manuscripts are currently under review, one from the Am J Clin Nutr and the other from J Immunol. Two additional manuscripts are being written and will be submitted in the next two months. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
We completed a randomized, double-blind, controlled crossover trial assessing the role of EPA and DHA on inflammation and lipid metabolism. Twenty-one subjects (9 men and 12 women, age 50-75 y) were enrolled and completed the study. Inclusion criteria were chronic inflammation (high sensitivity C reactive protein, hsCRP > 3 mg/L), elevated plasma triglyceride levels (TG, 90-500 mg/dL), and presence of at least one of the following: abdominal obesity (waist circumference > 102 cm in men and > 89 cm in women), hypertension (blood pressure >+ 130/80 mmHg or use of anti-hypertensive medications), and fasting plasma glucose >+ 100 mg/dL. The study consisted of a 4-week lead-in control phase (high oleic sunflower oil, 3 g/d) after which subjects were randomized to a 10-week supplementation phase with either EPA or DHA (3 g/d), followed by a 10-week washout phase, and then crossover. Blood was drawn at the end of the control and the EPA and DHA supplementation phases for the assessment of: a) serum cytokine levels, peripheral blood monocyte transcriptomics and inflammatory response to lipopolysaccharide (LPS) stimulation, and plasma lipid mediators derived from EPA and DHA, and b) plasma lipid and lipoprotein levels and activity of enzymes involved in lipoprotein metabolism. Regarding the effect of EPA and DHA on inflammation, there was no significant effect of these n-3 fatty acids on serum concentrations of hsCRP, tumor necrosis factor alpha (TNFα), interleukin 6 (IL6) and IL10, or monocyte chemoattractant protein 1 (MCP1). However, EPA significantly lowered the LPS-stimulated expression of TNFα and DHA significantly lowered the expression of TNFα, IL6 and MCP1, but also the anti-inflammatory IL10, in LPS-stimulated monocytes. Relative to EPA, DHA significantly lowered the expression of IL10. EPA supplementation was associated with significant increases in EPA-derived lipid mediators, including 18-hydoxyeicosapentaenoic acid (18-HEPE), the precursor of resolvins E1-3. DHA supplementation was associated with significant increases in both DHA-derived lipid mediators and EPA-derived lipid mediators. Principal component analysis (PCA) showed clear separation of the lipidome during EPA and DHA supplementation. The EPA-associated PCA component explained 44% of the reduction in LPS-stimulated monocyte TNFα gene expression during EPA supplementation and 19% of the reduction during DHA supplementation. The DHA-associated component did not explain TNFα expression. Analysis of monocyte transcriptome by RNA-sequencing revealed a clear sex dimorphism during the control phase, with expression of interferon-related genes higher in females than males. In addition, we observed a differential monocyte gene expression profile with EPA compared to DHA supplementation and a clear treatment by sex interaction. Regarding the effects of EPA and DHA on plasma lipid and lipoprotein metabolism, EPA significantly lowered plasma TG concentrations and cholesterol ester transfer protein (CETP) activity but did not change lipoprotein lipase (LPL) activity. On the other hand, DHA significantly reduced TG concentrations and increased LPL activity and LDL-C concentration without affecting CETP activity. The DHA-mediated increase in LPL activity was significantly correlated with the decrease in TG concentration. In conclusion, this clinical trial demonstrated differential effects of EPA and DHA on inflammation and lipid metabolism.
Publications
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2019
Citation:
So J, Wu D, Lichtenstein AH, Lamon-Fava S. Differential Effects of DHA and EPA Supplementation on Serum Inflammatory Markers and Blood Monocyte Inflammatory Response in Subjects with Chronic Inflammation. Vascular Biology. Boston , MA, May 14, 2019
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2019
Citation:
So J, Matthan MR, Maddipati KR, Lichtenstein AH, Wu D, Lamon-Fava S. Effects of EPA and DHA Supplementation on Plasma Specialized Proresolving Lipid Mediators and Blood Monocyte Inflammatory Response in Subjects with Chronic Inflammation. Nutrition 2019 (ASN), Baltimore, MD. June 11, 2019
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2019
Citation:
So J, Wu D, Lichtenstein AH, Lamon-Fava S. Docosahexaenoic Acid and Eicosapentaenoic Acid Supplementation Differentially Modulate Pro- and Anti-inflammatory Cytokines in Subjects with Chronic Inflammation. Nutrition 2019 (ASN), Baltimore, MD. June 11, 2019
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2020
Citation:
So J, Asztalos BF, Horvath KV, Lichtenstein AH, Lamon-Fava S. EPA and DHA have differential effects on cholesterol ester transfer protein and lipoprotein lipase activities following plasma triglycerides lowering. Nutrition 2020 (ASN), Seattle (virtual). June 1 2020
- Type:
Journal Articles
Status:
Under Review
Year Published:
2020
Citation:
So J, Wu D, Lichtentein AH, Tai AK, Matthan NR, Maddipati KR, Lamon-Fava S. Eicosapentaenoic acid and docosahexaenoic acid differentially modulate monocyte inflammatory response and plasma specialized pro-resolving mediators in subjects with chronic inflammation: a randomized, double-blind, crossover study. Am J Clin Nutr
- Type:
Journal Articles
Status:
Submitted
Year Published:
2020
Citation:
So J, Tai AK, Lichtenstein AH, Wu D, Lamon-Fava S. Sexual dimorphism in monocyte transcriptome in individuals with chronic low-grade inflammation. J Immunol
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2019
Citation:
So J, Matthan MR, Maddipati KR, Lichtenstein AH, Wu D, Lamon-Fava S. Effect of EPA and DHA Supplementation on Plasma Concentrations of Specialized Pro-resolving Lipid Mediators and their Association with Blood Monocyte Inflammatory Response in Subjects with Chronic Inflammation. Vascular Biology. Boston , MA, May 14, 2019
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Progress 02/15/18 to 02/14/19
Outputs
Target Audience:Target audience includes individuals with metabolic syndrome, individuals with elevated chronic inflammation and individuals with elevated plasma trigyceride levels. Changes/Problems:After changing some of the inclusion criteria to facilitate recruitment of subjects, we have not encountered problems in the enrollment or retention of study subjects. We do not expect technical problems in the assessment of our outcomes, as we have the necessary expertise. What opportunities for training and professional development has the project provided?The project has provided the opportunity to train one graduate student in the methodologies related to the objectives of the study. Specifically, the graduate student has been trained and performed the isolation of peripheral blood mononuclear cells (PBMC), the purification of a subtype of white blood cells called monocytes, the culture and stimulation of monocytes with lipopolysaccharide, the isolation of RNA, and the measurement of target messenger RNAs by real time PCR. The graduate students has also trained instatistical analysis. Based on preliminary results on the differential effects of EPA and DHA on monocyte inflammatory response and the production of specialized pro-resolving lipid mediators, abstracts have been submitted to the Nutrition 2019 conference (American Society for Nutrition) and to the Atherosclerosis Thrombosis and Vascular Biology/Vascular Discovery 2019 conference (American Heart Association). How have the results been disseminated to communities of interest?Abstracts on the the differentila effects of EPA and DHA supplementation on themonocyte inflammatory response and on the production of specialized pro-resolvong lipid mediators have been submitted to both the Nutrition 2019 conference and to the ATVB 2019 conference. What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, we plan to complete the following analyses on samples collected from all study participants, and write and publishthe related manuscripts: Differential effects of EPA and DHA on the transcriptomic profile of peripheral blood mononuclear cells by RNA-sequencing. Signaling pathways differentially affected by EPA and DHA in peripheral blood mononuclear cells. Effects of EPA and DHA on circulating pro-inflammatory M1-like and anti-inflammatory M2-like monocyte subtypes. Association of EPA- and DHA-derived pro-resolving lipid mediators with the expression of inflammatory genes and with monocyte phenotype (M1 vs M2). Lipid lowering effects of EPA and DHA: role of lipoprotein lipase and hepatic lipase on triglyceride lowering.
Impacts
What was accomplished under these goals?
We have completed the enrollment of subjects in this placebo-controlled, randomized, double-blind, crossover study assessing the effects of pure EPA and pure DHA, relative to a control oil (high oleic acid sunflower oil) on inflammation and lipid metabolism. Twenty-four subjects were enrolled into the study and 21 have completed the study. Three subjects have dropped out, one due to family reasons, one because he developed gout in the first week of his participation in the study (while he was on the placebo lead-in phase), and one developed leg cramps during the placebo lead-in phase of the study. We have completed the analysis of some of our proposed outcomes of inflammation and lipid metabolism, including blood monocyte inflammatory response, plasma cytokine concentrations,plasma concentrations ofEPA- and DHA-derived specialized pro-resolving lipid mediators and plasma lipid concentrations. We are currently working to complete all the proposed analyses, including the transcriptomic profile of peripheral blood mononuclear cells and pathway analysis.
Publications
- Type:
Conference Papers and Presentations
Status:
Submitted
Year Published:
2019
Citation:
So J, Wu D, Lichtenstein AH, Lamon-Fava S. Docosahexaenoic Acid and Eicosapentaenoic Acid Supplementation Differentially Modulate Pro- and Anti-inflammatory Cytokines in Subjects with Chronic Inflammation. Nutrition 2019 conference
- Type:
Conference Papers and Presentations
Status:
Submitted
Year Published:
2019
Citation:
So J, Matthan NR, Maddipati K, Lichtenstein AH, Wu D, Lamon-Fava S. Effects of EPA and DHA Supplementation on Plasma Specialized Proresolving Lipid Mediators and Blood Monocyte Inflammatory Response in Subjects with Chronic Inflammation. Nutrition 2019 conference
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Progress 02/15/17 to 02/14/18
Outputs
Target Audience:Target audience includes: individuals with metabolic syndrome, individuals with elevated chronic inflammation, individuals with elevated plasma lipid (triglyceride and cholesterol) levels, dieticians, physicians, researches in the fields of inflammation and lipid metabolism. Changes/Problems:During the previous funding period we had modified some inclusion criteria to facilitate the recruitment of study subjects. Since then, we have not encountered problems in the enrollment or retention of study subjects. We routinely communicate with our subjects over the phone every 4-5 weeks to increase communication with the study staff and increase subject retention. What opportunities for training and professional development has the project provided?Since 5 subjects still need to complete the study protocol,there has not yet been an opportunity to present resultsat aconference. Opportunities for professional development will materialize in the coming year. The project has provided the opportunity to train a graduate student in the methodologies that will be used to carry out the experiments related to the objectives of the study. Specifically, the graduate student has been trained in the isolation of peripheral blood mononuclear cells (PBMC),the purification of a subtype of white blood cells called monocytes, the culture and stimulation of monocytes with lipopolysaccharide, the isolation of monocyte RNA, and the measurement of target messenger RNAs by real time PCR. The PD, together with the student,is also developing flow cytometry protocols for the assessment of circulating monocyte subtypesand statistical analysis of RNA-sequencing data. How have the results been disseminated to communities of interest?No final results have yet been generated from this clinical trial, due to the fact that 5 study participant still need to complete thestudy. Results related to the specific objectives of the study are expected during this coming year.The results of this study will then be disseminated to the target audience via presentation at scientific meetings, publications in scientific journals, and outreach activities. What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, we expect that the 5 subjects currently enrolled in the study will complete the study. When all subjects have completed the study protocol, we willcarry out the following measurements in samples collected at the end of the placebo, EPA and DHA phases: 1) plasma inflammatory markers, transcriptomics of peripheral blood mononuclear cells, and monocyte inflammatory response,and2) plasma lipid levels andactivity of enzymes involved in lipid metabolism. To avoid variability, all samples from the same subject will be run in the same batch. The effects of EPA and DHA on these outcomes will be assessed bypre-specified statistical analysis.
Impacts
What was accomplished under these goals?
This placebo-controlled, randomized, double-blind, crossover study assesses the effects of pure EPA and pure DHA, relative to a control oil (high oleic acid sunflower oil) on inflammation and lipid metabolism.To date, we have pre-screened 405 subjects by phone interviews and 23 subjects have met all the inclusion and exclusion criteria and have been enrolled into the study. Three subjects have dropped out, one due to family reasons, one because he developed gout in the first week of his participation in the study (while he was on the placebo lead-in phase), and one developed leg cramps during the placebo lead-in phase of the study. Fifteen subjects have completed the 34-week study protocol and5 are currently in the study. All study subjects will complete their study participationby the Summer 2018. At that time, the final analysis of all inflammatory and lipid metabolism parameters will begin. The randomization code will become available when all laboratory analysis will be completed. Throughout the study, Dr. Lamon-Fava has met with Dr. Alice Lichtenstein (Co-Investigator), Dr. Edward Saltzman (Study Physician) and the recruiting and nursing services of the Metabolic Research Unit at the Jean Mayer USDA HNRCA to facilitate subject recruitment and retention, and to discuss adherence to study protocols. In addition, as detailed below, extensive training of a graduate student has occurred on several of the methodologies to be used to assess the study outcomes.
Publications
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Progress 02/15/16 to 02/14/17
Outputs
Target Audience:Target audience includes individuals with metabolic syndrome, individuals with elevated chronic inflammation, and individuals with elevated plasma triglyceride levels. Also, it includes dieticians and physicians. Changes/Problems:This studyhas arandomized, placebo-controlled, double-blind, crossover design, and was designed with a placebo lead-in phase lasting 4 weeks followed by randomization to either EPA or DHA for 12 weeks, then by a placebo phase of 12 weeks, andfinally by a crossover to either DHA or EPA for 12 weeks, for a total length of the study of 40 weeks.To increase study subjects' recruitment, retention and compliance, we have reduced the 12-week EPA, DHA and placebo phases to 10 weeks each, resulting in a total length of the study of 34 weeks. This change is not affecting the study design or subjects' safety,and still allows us to reliably measure the inflammatory andplasma lipidoutcomes as originally proposed in the grant, while enhancing feasibility and retention. In addition, we have changed one of the inclusion criteria of the study, namely plasma triglyceride levels,from 150-500 mg/dL to 90- 500 mg/dL. This change has increased the number of subjects eligible to enroll into the study. These changes to the original protocol have been approved by our Institutional Review Board (IRB). What opportunities for training and professional development has the project provided?The study started one year ago and there has not yet been an opportunity to present resultsat a scientificconference or as a scientific presentation. It is projected that opportunities for professional development will materialize in the coming year. The project has provided the opportunity to train a graduate student in the methodologies that will be used to carry out the experiments related to the objectives of the study. Specifically, the graduate student has been trained in the isolation of peripheral blood mononuclear cells (PBMC), purification of a subtype of white blood cells called monocytes, the culture and stimulation of monocytes with lipopolysaccharide, the isolation of RNA, and the measurement of target messenger RNAs by real time PCR How have the results been disseminated to communities of interest?This is a clinical trial and no results have been obtained yet. Results related to the specific objectives of the study are expected at the end of the study, when measurements of the outcomes of the study will be analyzed after unblinding of the treatment sequence in each subject. What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, we plan to enroll the remaining 12 subjects needed to complete the study. In addition, the following analyses on samples from the subjects who will have successfully completed the study protocol will be carried out: 1) plasma inflammatory markers, peripheral blood mononuclear cells expression of inflammatory genes and 2) plasma lipid levels, activity of enzymes involved in lipid metabolism.
Impacts
What was accomplished under these goals?
The study has a placebo-controlled, randomized, double bind, crossover design to assess the effects of pure EPA and pure DHA, relative to a control oil (high oleic acid sunflower oil) on inflammation and lipid metabolism. We have proposed to enroll 24 subjects in this study. Currently, we have pre-screened 153 subjects by phone interviews. Of these, 58 subjects were screened on-site for eligibility based on inclusion and exclusion criteria. Twelvesubjects (7 men and 5 women) have met all the criteria and have been enrolled into the study. Two subjects have dropped out, one due to family reasons and one because he developed gout in the first week of participation inthe study (while he was on the lead-in phase). Four subjects have already completed the 34 week study protocol. Throughout the study, from its start until now, there have been frequent meetings of the Principal Investigator with the study personnel, including Dr. Alice Lichtenstein (Co-Investigator), Dr. Edward Saltzman (Study Physician) and the recruiting and nursing services of the Metabolic Research Unit to facilitate study initiation, subject recruitment and retention, and to discuss adherence to study protocols. In addition, as detailed below, extensive training of a graduate student has occurred on several of the methodologies to be used to assess outcomes.
Publications
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Progress 02/15/15 to 02/14/16
Outputs
Target Audience:Target audience includes individuals with metabolic syndrome, individuals with elevated chronic inflammation, individuals with elevated triglyceride levels. It also includes dietitians and physicians. Changes/Problems:The original studyhas arandomized, placebo-controlled, double-blind, crossover designand consists of a placebo lead-in phase lasting 4 weeks followed by randomization to either EPA or DHA for 12 weeks, then by a placebo phase of 12 weeks, andfinally by a crossover to either DHA or EPA for 12 weeks, for a total length of the study of 40 weeks.To increase study subjects' recruitment, retention and compliance, we arereducing the 12-week EPA, DHA and placebo phases to 10 weeks each, resulting in a total length of the study of 34 weeks. This change is not affecting the study design or subjects' safety,and still allows us to reliably measure the inflammatory andplasma lipidoutcomes as originally proposed in the grant, while enhancing feasibility and retention. In addition, we propose to change one of the inclusion criteria of the study, namely plasma triglyceride levels,from 150-500 mg/dL to 90-500 mg/dL. This change will increase the number of subjects eligible to enroll into the study. We have experienced great difficulty in recruiting subjects in astudy with similar inclusion criteria and therefore want to use this experience to improve the recruitment and enrollment of subjects in the current study. What opportunities for training and professional development has the project provided?The project has provided the opportunity to train a graduate student in the methodologies that will be used to carry out experiments related to the objectives of the study. Specifically, the graduate student has been trained in the isolation of peripheral blood mononuclear cells (PBMC), purification of a subtype of white blood cells called monocytes, the culture and stimulation of monocytes with lipopolysaccharide, the isolation of RNA, and the measurement of target messenger RNAs by real time PCR. How have the results been disseminated to communities of interest?No results have been obtained because of the delay in the start of the study. What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, I plan to screen over 100 subjects to enroll 10-12 subjects into the randomized crossover study. I will carry out the study procedures for all the enrolled subjects who have completed the study, and I will attempt to enhance the overall adherence to the protocol by participating subjects.
Impacts
What was accomplished under these goals?
The study is still in the early recruiting phase because there were unexpected delays in the start of the study. While Institutional Review Board (IRB) approval for the study was obtained within one month of the start day of the project (March 10, 2015), theomega-3 supplements, namely pure eicosapentaenoic acid (EPA) and pure docosahexaenoic acid (DHA), were obtained only on January 6, 2016, when they were shipped to Tufts University by the manufacturer. The delay in obtaining the omega-3 supplements was due to lengthy manufacturing processes requiring extensive purification to ensure purity of the EPA and DHA supplements. Gas chromatography of the supplements, as provided by the manufacturing company, show that the EPA supplements contain 761 mg EPA per capsule without any DHA, while the DHA supplements contain 770 mg DHA + 3 mg EPA per capsule, therefore reaching a greater than 95% purity, as specified in the grant proposal. No other product with such characteristics of purity is currently commercially available. From the start day of the project until now, there have been frequent meetings of the Principal Investigator with the study personnel, including Dr. Alice Lichtenstein (Co-Investigator), Dr. Edward Saltzman (Study Physician) and the recruiting and nursingpersonnel of the Metabolic Research Unit to facilitate study initiation and develop protocols forsample collection, handling and storage. In addition, as detailed below, extensive training of a graduate student has occurred on several of the methodologies to be used to assess outcomes. In addition, the PI has also finalized the manual of procedures, capturing all the critical aspects of the implementation of the study.
Publications
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