Epigenetic Analysis of Regulation of the Inflammasome-activating NLRP3 Gene in Monocytes From Atrial Fibrillation Patients and Controls

Study Overview
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Brief Summary
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The study will analyze blood from volunteers to determine whether there is an underlying epigenetic cause of the inflammation of the heart associated with atrial fibrillation (AF) and its progression with age. Confirming the regions of epigenetic elements associated with upregulation of the inflammatory genes will help the investigators in identifying target sites for developing future therapeutic interventions. The investigators propose to confirm the monocyte-cell type specific DNA methylation profile of NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) and determine the age-related and AF-related changes in DNA methylation and expression of NLRP3 in monocytes. This study will provide insights into the epigenetic regulation of NLRP3 in young and elderly patients, as well as in AF patients vs. controls which will help in devising methods of modulating NLRP3 expression and decreasing cardiac fibrosis progression.

Detailed Description
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Blood samples will be collected from 120 female participants as follows: 10 women between the ages of 50 - 80 years diagnosed with AF as evidenced by rhythm strips or written documentation; 10 healthy women subjects between the ages of 50-80 years and 10 healthy women between the ages of 20-30 years. Participants will be recruited from the Tulane University Cardiology Clinics and also from volunteer research registries. These participants will be asked for consent to draw for another 8 ml (1.5 teaspoons) of blood during routine diagnostic blood draws. These blood samples will be de-identified by the PI upon receipt and no identifiable information will be maintained. Leukocyte subtypes including monocytes, neutrophils and B cells will be isolated from blood and DNA extraction will be carried out from the leukocyte subtypes. These leukocyte DNA samples will be sent to New England Biolabs, Ipswich, Massachusetts for DNA methylation analysis to identify target regions of epigenetic elements associated with upregulation of NLRP3 gene expression that might be related to disease progression with age. Real-Time Polymerase chain reaction (PCR) will be carried out at Tulane Research and Innovation for Arrhythmia Discoveries (TRIAD) Center, Tulane University School of Medicine from RNA extracted from leukocyte subtype to check the level of expression of NLRP3 highly associated with inflammation in AF.

Official Title
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Epigenetic Analysis of Regulation of the Inflammasome-activating NLRP3 Gene in Monocytes From Atrial Fibrillation Patients and Controls

Conditions
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Atrial Fibrillation, Arrhythmia

Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Subjects aged 18 or above, Control Group scheduled for routine annual health check-up and healthy blood donors without any documentation of cardiovascular disease or other serious immune-related illnesses, such as diabetes, hypertension, autoimmune arthritis etc healthy women subjects between the ages of 50-80 years healthy women subjects between the ages of 20-30 years. Study Patients -women between the ages of 50 - 80 years diagnosed with AF as evidenced by rhythm strips or written documentation. Exclusion Criteria: Previous left atrial ablation or any type of valvular surgery Patients who are taking medications like Ibuprofen, Toradol, Naproxen and other common over-the -counter NSAIDs. Women who are pregnant Terminally ill patients

Ages Eligible for Study
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Minimum Age: 20 Years
Maximum Age: 80 Years

Sexes Eligible for Study
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Female

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Patient Group<br>10 women between the ages of 50 - 80 years diagnosed with AF as evidenced by rhythm strips or written documentation. | |
| Control Group 1<br>10 healthy women subjects between the ages of 50-80 year | |
| Control Group 2<br>10 healthy women subjects between the ages of 20-30 years | |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Determine whether there is an underlying epigenetic cause of the inflammation of the heart associated with atrial fibrillation | This will be determined by confirming the monocyte-cell type specific DNA methylation profile of NLRP3. | Day 1 | 
| Determine whether there are significant associations of DNA methylation with NLRP3 expression in monocytes with advancing age. | Age association will be examined by comparing group wise the DNA methylation levels of monocytes from young vs. old individuals. The level of NLRP3 expression will be compared to the levels of DNA methylation. | Day 1 | 
| Determine whether there are significant associations of DNA methylation with NLRP3 expression in monocytes with the status of atrial fibrillation. | Disease associations will be tested by comparing group wise the DNA methylation levels of monocytes from AF patients vs. controls. The investigators will also test for understanding significant association of NLRP3 expression with corresponding DNA methylation status in AF patients. | Day 1 | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Inflammation, Stroke, DNA methylation