RNA Cloning and Visualization in Human Atherosclerosis

This study is ongoing, but not recruiting participants.
Sponsor:
Information provided by (Responsible Party):
Bradley Gelfand, PhD, University of Kentucky
ClinicalTrials.gov Identifier:
NCT01928095
First received: August 20, 2013
Last updated: October 21, 2013
Last verified: October 2013
  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: Time Perspective: Prospective
Official Title: RNA Cloning and Visualization in Human Atherosclerosis

Resource links provided by NLM:


Further study details as provided by University of Kentucky:

Primary Outcome Measures:
  • Altered Dicer Activity [ Time Frame: 1 year ] [ Designated as safety issue: No ]
    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 ] [ Designated as safety issue: No ]
    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 ] [ Designated as safety issue: No ]
    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


Estimated Enrollment: 20
Study Start Date: October 2013
Estimated Study Completion Date: November 2015
Estimated Primary Completion Date: November 2015 (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

Ages Eligible for Study:   18 Years and older
Genders Eligible for Study:   Both
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
Please refer to this study by its ClinicalTrials.gov identifier: NCT01928095

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

No publications provided

Responsible Party: Bradley Gelfand, PhD, Principal Investigator, University of Kentucky
ClinicalTrials.gov Identifier: NCT01928095     History of Changes
Other Study ID Numbers: IRB # 13-0623-F1V
Study First Received: August 20, 2013
Last Updated: October 21, 2013
Health Authority: United States: Institutional Review Board

Keywords provided by University of Kentucky:
atherosclerosis
carotid endartherectomy

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

ClinicalTrials.gov processed this record on April 16, 2014