"Immunoregulation in Atherosclerosis: A Single-Cell RNA Sequencing Study"

NCT ID: NCT06860295

Last Updated: 2025-06-10

Basic Information

• Recruitment Status: NOT_YET_RECRUITING
• Total Enrollment: 30 participants
• Study Classification: OBSERVATIONAL
• Study Start Date: 2025-06-25
• Study Completion Date: 2027-04-25

Brief Summary

Atherosclerosis is the leading cause of acute cardiovascular events, such as myocardial infarction and stroke, and is a significant risk factor for cardiovascular mortality. The detailed understanding of the immune mechanisms and cellular transformations involved in the pathogenesis of atherosclerosis is still limited, and the use of single-cell RNA sequencing (scRNAseq) has revealed new cellular functions and subpopulations associated with disease progression. This study aims to identify cellular subpopulations, molecular pathways, and changes in gene expression related to the development of atherosclerosis in human coronary arteries. Using scRNAseq, the study seeks to characterize the transcriptomic landscape of cells present in atherosclerotic plaques and identify molecular signatures that reveal individual predispositions to specific phenotypes, such as disease susceptibility and response to therapies. The research will be conducted at the Albert Einstein Israeli Hospital in São Paulo and will involve samples from coronary arteries and atherosclerotic plaques of the explanted hearts of patients who have undergone heart transplants as well as from discarded material of coronary artery bypass graft surgery (CABG). With an estimated sample size of 20-30 plaques, the data obtained will allow for a detailed analysis of the molecular mechanisms involved in atherosclerosis, contributing to the development of specific therapeutic targets.

Detailed Description

This is a prospective observational study that will be conducted with samples of coronary arteries and atherosclerotic plaques from the explanted hearts of patients with ischemic cardiomyopathy who have undergone heart transplants as well as from discarded material of coronary artery bypass graft surgery . The atherosclerotic plaque samples will be retrieved from the coronary arteries during the transplant surgery or the CABG procedure and transported to the laboratory within 2 hours. The selected arteries will be dissected and microscopically evaluated, and the samples will be fixed in formalin, embedded in paraffin, and stained for histological analysis. The tissue will be enzymatically dissociated to obtain individual cells, which will then be filtered and assessed for cell viability. The cells will undergo single-cell RNA sequencing (scRNAseq) with the aim of analyzing around 5,000-10,000 cells per sample. The sample size will be 20-30 plaques, with the goal of capturing the main cellular populations and performing a robust analysis.

Conditions

Atherosclerosis of Coronary Artery

Study Design

• Observational Model Type: CASE_ONLY
• Study Time Perspective: PROSPECTIVE

Study Groups

Coronary Plaques from Explanted Heart

The selected arteries will be dissected and microscopically evaluated, and the samples will be fixed in formalin, embedded in paraffin, and stained for histological analysis. The tissue will be enzymatically dissociated to obtain individual cells, which will then be filtered and assessed for cell viability. The cells will undergo single-cell RNA sequencing (scRNAseq) with the aim of analyzing around 5,000-10,000 cells per sample. The sample size will be 20-30 plaques, with the goal of capturing the main cellular populations and performing a robust analysis.

Single Cell RNA Sequencing

Intervention Type: DIAGNOSTIC_TEST

Reprocessing

Data normalization, dimensionality reduction, and clustering will be performed using the Seurat package (based on R).

Cell types will be identified based on canonical marker gene expression. Cell Subpopulation Identification

Clusters will be annotated based on signatures of known cell types (e.g., endothelial cells, smooth muscle cells, macrophages, etc.).

Differential gene expression analysis will be conducted to identify disease-associated genes.

Pathway and Function Enrichment

Gene Set Enrichment Analysis (GSEA) and pathway analyses (e.g., KEGG, Reactome) will be carried out to investigate the biological processes and pathways driving atherosclerosis.

Quality Control

Sequencing reads will be assessed for quality using FASTQC, and low-quality reads will be filtered out.

The Cell Ranger software (10x Genomics) will be used to align reads to the human genome and quantify gene expression at the single-cell level.

Interventions

Single Cell RNA Sequencing

Reprocessing

Data normalization, dimensionality reduction, and clustering will be performed using the Seurat package (based on R).

Cell types will be identified based on canonical marker gene expression. Cell Subpopulation Identification

Clusters will be annotated based on signatures of known cell types (e.g., endothelial cells, smooth muscle cells, macrophages, etc.).

Differential gene expression analysis will be conducted to identify disease-associated genes.

Pathway and Function Enrichment

Gene Set Enrichment Analysis (GSEA) and pathway analyses (e.g., KEGG, Reactome) will be carried out to investigate the biological processes and pathways driving atherosclerosis.

Quality Control

Sequencing reads will be assessed for quality using FASTQC, and low-quality reads will be filtered out.

The Cell Ranger software (10x Genomics) will be used to align reads to the human genome and quantify gene expression at the single-cell level.

Intervention Type: DIAGNOSTIC_TEST

Eligibility Criteria

Inclusion Criteria

• Patients diagnosed with ischemic cardiomyopathy due to advanced atherosclerosis, undergoing heart transplantation;
• Patients with coronary artery disease submitted to coronary artery bypass graft surgery (CABG)."
• Aged between 40 and 75 years;
• Signed informed consent form.

Exclusion Criteria

• Patients with systemic inflammatory or autoimmune diseases;
• History of cancer within the last 5 years or active malignancy;
• Recent use (within the last 6 months) of immunosuppressive therapy.

• Minimum Eligible Age: 45 Years
• Maximum Eligible Age: 75 Years
• Eligible Sex: ALL
• Accepts Healthy Volunteers: No

Sponsors

Hospital Israelita Albert Einstein

OTHER

Sponsor Role: lead

Responsible Party

Carlos Vicente Serrano Junior

Principal Investigator

Responsibility Role: PRINCIPAL_INVESTIGATOR

Locations

Hospital Israelita Albert Einstein

São Paulo, São Paulo, Brazil

Countries

Brazil

Central Contacts

Carlos Vicente Serrano, PhD

Role: CONTACT

carlos.sjunior@einstein.br

+55 (11) 2151-5408

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Other Identifiers

AteroImunoHIAE

Identifier Type: -

Identifier Source: org_study_id