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RNA Cloning and Visualization in Human Atherosclerosis

This study has been completed.
Sponsor:
ClinicalTrials.gov Identifier:
NCT01928095
First Posted: August 23, 2013
Last Update Posted: April 13, 2017
The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Listing a study does not mean it has been evaluated by the U.S. Federal Government. Read our disclaimer for details.
Information provided by (Responsible Party):
Bradley Gelfand, PhD, University of Virginia
  Purpose

The research objectives of this project are as follows:

  1. Obtain high-quality human atherosclerotic arterial samples from diseased donors.
  2. Perform biochemical analysis to determine the abundance, localization and activity of Dicer and double-stranded RNAs in these diseased tissues.

Condition
Atherosclerosis

Study Type: Observational
Study Design: Observational Model: Other
Time Perspective: Prospective
Official Title: RNA Cloning and Visualization in Human Atherosclerosis

Resource links provided by NLM:


Further study details as provided by Bradley Gelfand, PhD, University of Virginia:

Primary Outcome Measures:
  • Altered Dicer Activity [ Time Frame: 1 year ]
    The investigator will measure the relative levels of abundant canonical Dicer substrates/enzymatic products (i.e. the ratio of pre- to mature micro-RNA) via northern blot


Secondary Outcome Measures:
  • Are downstream mediators of Dicer dysregulation activated [ Time Frame: 1 Year ]
    Analysis will compare levels and activation of the inflammasome (caspase-1, NLRP3, ASC), cytokines (IL-1β, IL-18) and signaling intermediates (MyD88, IRAK1/4) between healthy and diseased tissues.


Other Outcome Measures:
  • Dicer dysregulation displays cell type-specific patterns. [ Time Frame: 1 year ]
    The investigator will co-stain tissue sections with antibodies recognizing Dicer as well as cell-specific markers (CD31 for endothelium, SMA for smooth muscle, MAC-1 for macrophages)


Biospecimen Retention:   Samples Without DNA
discarded artherosclerotic tissue obtained during carotid endarterectomy

Enrollment: 22
Study Start Date: October 2013
Study Completion Date: November 2016
Primary Completion Date: November 2016 (Final data collection date for primary outcome measure)
Groups/Cohorts
Carotid Endocardectomy Patients
Subsequent analyses will be performed on the basis of the pathological grading provided by UK Pathology (e.g do readouts of the Dicer pathway correlate with pathological plaque characteristics).

Detailed Description:

Participants enrolled in to this pilot study will not be randomized and will not receive any investigation medication.

In collaboration with Dr. Sibu Saha, Professor of Surgery, University of Kentucky, the investigator will obtain freshly isolated human atherosclerotic tissue from patients undergoing carotid endarterectomy. The surgical process by which the tissue is obtained is not part of this research project. These tissues are surgically removed from patients as a treatment for atherosclerosis of the carotid arteries and typically sent for pathological examination.

After the the clinical pathology has been completed, the investigator will obtain and analyze a small amount of discarded (200mg) tissue. Subsequent analyses will be performed on the basis of the pathological grading provided by UK Pathology (e.g do readouts of the Dicer pathway correlate with pathological plaque characteristics). Tissue will be anonymized, with only information with respect to drug history, age, gender and pathological grade of the tissue.

The will extract protein and RNA. Dicer abundance will be assessed by western blot, quantitative RT-PCR and northern blot. Dicer related RNAs will be measured by quantitative RT-PCR and northern blot. The invesigator will inspect the tissue for evidence of altered Dicer activity by measuring the relative levels of abundant canonical Dicer substrates/enzymatic products (i.e. the ratio of pre- to mature micro-RNA) via northern blot. The investigator predicts that Dicer levels and activity are reduced in human atherosclerotic tissue compared to healthy arteries.

Next, the investigator will assess whether downstream mediators of Dicer dysregulation are activated in these tissue samples by comparing levels and activation of the inflammasome (caspase-1, NLRP3, ASC), cytokines (IL-1β, IL-18) and signaling intermediates (MyD88, IRAK1/4) between healthy and diseased tissues. The investigator predicts that atherosclerotic plaques will exhibit evidence of Dicer dysregulation.

Next, the investigator will co-stain tissue sections with antibodies recognizing Dicer as well as cell-specific markers (CD31 for endothelium, SMA for smooth muscle, MAC-1 for macrophages) to assess whether Dicer dysregulation displays cell type-specific patterns.

  Eligibility

Information from the National Library of Medicine

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Ages Eligible for Study:   18 Years and older   (Adult, Senior)
Sexes Eligible for Study:   All
Accepts Healthy Volunteers:   No
Sampling Method:   Probability Sample
Study Population
Male and Female patients undergoing carotid endarterectomy
Criteria

Inclusion Criteria:

  • Age of 18 and older
  • Undergoing carotid endarterectomy

Exclusion Criteria:

  • Under 18 years of age
  • Not undergoing carotid endarterectomy
  Contacts and Locations
Information from the National Library of Medicine

To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.

Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT01928095


Locations
United States, Kentucky
University of Kentucky Chandler Medical Center
Lexington, Kentucky, United States, 40536
Sponsors and Collaborators
University of Virginia
Investigators
Principal Investigator: Bradley Gelfand, PhD University of Virginia, Department of Ophthalmology
Principal Investigator: Bradley Gelfand, PhD Ophthalmology
  More Information

Responsible Party: Bradley Gelfand, PhD, Principal Investigator, University of Virginia
ClinicalTrials.gov Identifier: NCT01928095     History of Changes
Other Study ID Numbers: IRB # 13-0623-F1V
First Submitted: August 20, 2013
First Posted: August 23, 2013
Last Update Posted: April 13, 2017
Last Verified: April 2017

Studies a U.S. FDA-regulated Drug Product: No
Studies a U.S. FDA-regulated Device Product: No

Keywords provided by Bradley Gelfand, PhD, University of Virginia:
atherosclerosis
carotid endartherectomy

Additional relevant MeSH terms:
Atherosclerosis
Arteriosclerosis
Arterial Occlusive Diseases
Vascular Diseases
Cardiovascular Diseases