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

Official title

The Architectural Immunoregulation in Atherosclerotic Disease: A Single-Cell RNA Sequencing and Spatial Biology Approach

Overview

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.

Interventions

• Diagnostic test 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 Enrichme

Primary outcome measures

• Concentration of Specific Cells Clusters And Macrophage / Linfocite Subpopulations : Create a high-resolution single-cell atlas of atherosclerotic plaques. [Time frame: Through study completion, an average of 1 year and a half.]

Secondary outcome measures (1)

• Concentration of interleukins and cellular subtypes by citometer flow to Identification of molecular signatures in atherosclerosis [Time frame: Through study completion, an average of 1 year and a half .]

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.

Criteria are shown verbatim from the registry (in English). Final eligibility is always assessed by the study center.

Healthy volunteers: No

Study design

Observational model

Case-only

Study locations

Brazil · 1 center

• Hospital Israelita Albert Einstein — São Paulo

Publications

• Zhang H, Viveiros A, Nikhanj A, Nguyen Q, Wang K, Wang W, Freed DH, Mullen JC, MacArthur R, Kim DH, Tymchak W, Sergi CM, Kassiri Z, Wang S, Oudit GY. The Human Explanted Heart Program: A translational bridge for cardiovascular medicine. Biochim Biophys Acta Mol Basis Dis. 2021 Jan 1;1867(1):165995. doi: 10.1016/j.bbadis.2020.165995. Epub 2020 Oct 22. PMID 33141063
• Rajab TK, Singh SK. Donation After Cardiac Death Heart Transplantation in America Is Clinically Necessary and Ethically Justified. Circ Heart Fail. 2018 Mar;11(3):e004884. doi: 10.1161/CIRCHEARTFAILURE.118.004884. No abstract available. PMID 29664408
• Price AL, Patterson N, Yu F, Cox DR, Waliszewska A, McDonald GJ, Tandon A, Schirmer C, Neubauer J, Bedoya G, Duque C, Villegas A, Bortolini MC, Salzano FM, Gallo C, Mazzotti G, Tello-Ruiz M, Riba L, Aguilar-Salinas CA, Canizales-Quinteros S, Menjivar M, Klitz W, Henderson B, Haiman CA, Winkler C, Tusie-Luna T, Ruiz-Linares A, Reich D. A genomewide admixture map for Latino populations. Am J Hum Gen PMID 17503322
• Gouveia MH, Bentley AR, Leal TP, Tarazona-Santos E, Bustamante CD, Adeyemo AA, Rotimi CN, Shriner D. Unappreciated subcontinental admixture in Europeans and European Americans and implications for genetic epidemiology studies. Nat Commun. 2023 Nov 7;14(1):6802. doi: 10.1038/s41467-023-42491-0. PMID 37935687
• Hu Z, Liu W, Hua X, Chen X, Chang Y, Hu Y, Xu Z, Song J. Single-Cell Transcriptomic Atlas of Different Human Cardiac Arteries Identifies Cell Types Associated With Vascular Physiology. Arterioscler Thromb Vasc Biol. 2021 Apr;41(4):1408-1427. doi: 10.1161/ATVBAHA.120.315373. Epub 2021 Feb 25. PMID 33626908
• Soehnlein O, Libby P. Targeting inflammation in atherosclerosis - from experimental insights to the clinic. Nat Rev Drug Discov. 2021 Aug;20(8):589-610. doi: 10.1038/s41573-021-00198-1. Epub 2021 May 11. PMID 33976384
• Fernandez DM, Rahman AH, Fernandez NF, Chudnovskiy A, Amir ED, Amadori L, Khan NS, Wong CK, Shamailova R, Hill CA, Wang Z, Remark R, Li JR, Pina C, Faries C, Awad AJ, Moss N, Bjorkegren JLM, Kim-Schulze S, Gnjatic S, Ma'ayan A, Mocco J, Faries P, Merad M, Giannarelli C. Single-cell immune landscape of human atherosclerotic plaques. Nat Med. 2019 Oct;25(10):1576-1588. doi: 10.1038/s41591-019-0590-4 PMID 31591603
• Wirka RC, Wagh D, Paik DT, Pjanic M, Nguyen T, Miller CL, Kundu R, Nagao M, Coller J, Koyano TK, Fong R, Woo YJ, Liu B, Montgomery SB, Wu JC, Zhu K, Chang R, Alamprese M, Tallquist MD, Kim JB, Quertermous T. Atheroprotective roles of smooth muscle cell phenotypic modulation and the TCF21 disease gene as revealed by single-cell analysis. Nat Med. 2019 Aug;25(8):1280-1289. doi: 10.1038/s41591-019-05 PMID 31359001

Identifiers

NCT: NCT06860295 · AteroImunoHIAE

Primary sources (government registries)

• ClinicalTrials.gov (US NIH)

View this study on ClinicalTrials.gov ↗