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

NCT06860295 | Not Yet Recruiting

• 1 other identifier: AteroImunoHIAE
• Study Type: observational
• Target: 30
• Locations: 1 country
• Sites: 1

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.

Conditions

Atherosclerosis of Coronary Artery

Keywords

Atherosclerosis; Single Cell RNA-sequencing; Immunology

Outcome Measures

Primary Outcomes (1)

• Concentration of Specific Cells Clusters And Macrophage / Linfocite Subpopulations : Create a high-resolution single-cell atlas of atherosclerotic plaques.

  Determine the concentration of cellular clusters with specific inflammatory subpopulations determination to Create a high-resolution single-cell atlas of atherosclerotic plaques, revealing cell-specific transcriptional changes associated with the disease. The findings could identify new therapeutic targets for intervention in atherosclerosis and enhance the understanding of its molecular pathogenesis.

  Through study completion, an average of 1 year and a half.

Secondary Outcomes (1)

• Concentration of interleukins and cellular subtypes by citometer flow to Identification of molecular signatures in atherosclerosis

  Through study completion, an average of 1 year and a half .

Study Arms (1)

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.

Diagnostic Test: Single Cell RNA Sequencing

Interventions

Single Cell RNA Sequencing 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.

Coronary Plaques from Explanted Heart

Eligibility Criteria

• Age: 45 Years - 75 Years
• Sex: all
• Healthy Volunteers: No
• Age Groups: Adult (18-64), Older Adult (65+)
• Sampling Method: Probability Sample

Study Population

Patients diagnosed with ischemic cardiomyopathy due to advanced atherosclerosis, undergoing heart transplantation.

You may qualify if:

• 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.

You may not qualify if:

• 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.

Sponsors & Collaborators

• Hospital Israelita Albert Einstein: lead

Study Sites (1)

Hospital Israelita Albert Einstein

São Paulo, São Paulo, 05652-900, Brazil

Location

Related Publications (15)

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Biospecimen

Retention: SAMPLES WITH DNA

Tissue Dissociation The tissue from the plates will be enzymatically dissociated into single-cell suspensions using a combination of collagenase, DNase I, and dispase. Cells will be filtered through a 70 µm filter to remove debris and aggregates. Viability Check and Enrichment Cell viability will be assessed using Trypan blue exclusion. Dead cells and debris will be removed using a magnetic kit for dead cell removal, if necessary. scRNAseq Workflow Library Preparation Single-cell capture and library preparation will be performed using a droplet-based microfluidic system (e.g., 10x Genomics Chromium). Approximately 5,000-10,000 cells per sample will be targeted for analysis.

MeSH Terms

Conditions

Coronary Artery Disease; Atherosclerosis

Condition Hierarchy (Ancestors)

Coronary Disease; Myocardial Ischemia; Heart Diseases; Cardiovascular Diseases; Arteriosclerosis; Arterial Occlusive Diseases; Vascular Diseases

Central Study Contacts

Carlos Vicente Serrano, PhD

CONTACT

+55 (11) 2151-5408; carlos.sjunior@einstein.br

Study Design

• Study Type: observational
• Observational Model: CASE ONLY
• Time Perspective: PROSPECTIVE
• Target Duration: 2 Years
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Principal Investigator

Study Record Dates

• First Submitted: February 24, 2025
• First Posted: March 6, 2025
• Study Start: June 25, 2025
• Primary Completion (Estimated): October 25, 2026
• Study Completion (Estimated): April 25, 2027
• Last Updated: June 10, 2025

Record last verified: 2025-06

Data Sharing

• IPD Sharing: Will share
• Shared Documents: STUDY PROTOCOL, SAP, CSR

Locations

BR (1)