A Review of Macrophage Diversity and Immune Checkpoint Regulation in Atherosclerosis: Insights from Single-Cell Transcriptomes

A Review of Macrophage Diversity and Immune Checkpoint Regulation in Atherosclerosis: Insights from Single-Cell Transcriptomes | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Systematic Review A Review of Macrophage Diversity and Immune Checkpoint Regulation in Atherosclerosis: Insights from Single-Cell Transcriptomes Hossein Nasouti Aghjehroud, Roghayeh Alizadeh, Aryan Mahmood Faraj, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7393034/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Purpose of Review: In this research, the idea of ​​single cell transcription was used to describe atherosclerotic plaque macrophages. Recent Findings: The results of the single-cell studies provided detailed data on the transcriptional and phenotypic diversity of macrophages in plaques. Macrophages are regulated by a variety of factors, including cell interactions, oxygen levels, nutrients, metabolites or other soluble signals, and (combination or alteration of) the extracellular matrix. These signals can influence macrophage selection and survival, as well as differentiation and polarization. In addition, macrophages in these plaques were replaced by activated phenotypes containing subsets associated with plaque vulnerability. Summary: Atherosclerosis is due to the multifaceted contribution of the immune system to the traffic and vascular homes. However, the specific properties of altered immune cells in atherosclerotic lesions that lead to clinical events such as ischemic stroke and myocardial infarction are not well-defined. Therefore, our approach provides a powerful tool to help study the underlying mechanisms of human diseases and discover novel pharmacological approaches for therapeutic interventions. Atherosclerosis Various macrophages Single cell transcription Phenotypic diversity myocardial infarction Figures Figure 1 Figure 2 Figure 3 Figure 4 1. Introduction Atherosclerosis (AS) is a complex metabolic and inflammatory disease of the arterial wall, characterized by the accumulation of lipoproteins rich in cholesterol and inflammatory cells. Accumulation of lipoproteins containing lipoproteins containing apolipoprotein B Atherosclerosis is the main cause of cardiovascular events such as myocardial infarction and stroke, resulting in 17.9 million deaths worldwide [ 1 ]. The progression of the atherosclerotic plate is preferred due to hypercholesterolemia and dyslipidemia. Its development begins when macrophages living in the inner membrane occupy excess lipids and form bubble cells. Over the past 20 years, achievements in the field of atherosclerosis have identified myeloidemia, the proliferation of plaque and steatosis macrophages as an important determinant of atherosclerosis development and regression. These parameters have recently been shown to be affected by immune cell metabolism [ 2 – 4 ]. It is interesting to note that while the clearance of lipoproteins by macrophages plays a similar role in preventing the production of harmful lipids, excessive lipid metabolism in macrophages leads to phenotypic and function changes, ultimately leading to death [ 5 ]. Macrophages, essential immune cells, are truly known for their remarkable ability to accept a variety of phenotypes and functions depending on the environment, and recently, sequencing of single-cell RNA RNAs has significantly increased our understanding of macrophage heterogeneity in atherosclerotic zone classification [ 6 ]. This technique provides a more detailed description of these cells and their role in the potential progression and treatment of atherosclerosis. SCRNA-EQ provides molecular profiles to individual macrophages with high resolution, allowing researchers to identify unique gene expression models and derive specific functions of various water macrophages in atherosclerotic lesions [ 7 – 9 ]. As a result, understanding the phenotypic complexity of macrophages is important for developing effective therapeutic strategies. Aiming for modulation of a specific subset of macrophages or their function satisfies a perspective to prevent or modify plate progression. Because macrophages play an important role in all stages of atherosclerosis and are involved in inflammation, lipid accumulation and plate evolution are important for different subpopulations to develop targeted therapies. As a result, this study aims to discuss the regulation of various macrophages and immune control points in atherosclerosis. This discussion overturned unprecedented light on the transcriptional environment and phenotypic heterogeneity of atherosclerotic macrophages, exhibited special functions, and opened up new opportunities to study different populations and their functions in different populations of atherosclerosis. 2. Research Methodology The design of the study was a review. This study looked for and investigated articles related to various macrophages of atherosclerosis. Particularly, we understood the base using PubMed, Web of Science (clariivate Analytics), Scopus, and Kokranée libraries and the appropriate keywords, and were associated with a variety of macrophages in atherosclerosis. The study was conducted in June 2025 and approved by the counterpart council. We are also considering Google Scholar to find the right item. It also includes a list of article links. 3. Progression of atherosclerosis Atherosclerosis has been increasing over the years through various stages of endothelial damage, the formation of fat bands, plate growth, rupture of plates or erosion, causing potential complications such as heart attacks and strokes [ 10 ]. This process is universal, but accelerates risk factors such as high cholesterol, high blood pressure, diabetes, smoking and obesity (Fig. 1 ). This process begins with damage to the internal mucosa of the artery (endothelium). This causes an immune response in which the white blood is replenished in the field. LDL cholesterol (bad cholesterol) accumulates and is associated with damaged endothelium, forming early deposits called fatty stripes [ 11 ]. These fat bands develop in more complex plates consisting of cholesterol, fatty substances, calcium and cell waste. Fibrous coverage is formed on the plate and can be stabilized or unstable [ 12 ]. The plate can explode, which leads to the formation of a blood clot (thrombosis). This thrombus can seriously or completely block the artery and, if it affects the brain, affecting the heart or stroke, causes a heart attack [ 13 ]. On the other hand, macrophages rapidly recognize and collect LDL-C via surface binge, and convert them into cells to form the first atherosclerotic lesions [ 14 ]. However, it becomes clear that lipid reduction alone does not completely interfere with the progression of atherosclerosis, revealing that macrophages are new potential therapeutic targets. 3.1. Epidemiology of atherosclerosis Atherosclerosis primarily affects the heart and brain, manifesting as coronary artery disease and ischemic stroke, and according to a 2019 epidemiological study, there are places that cannot be denied in the choice of mortality around the world [ 15 ]. The World Health Organization has shown that coronary arteries and stroke together account for around 85% of deaths, exceeding other causes, including cancer [ 16 ]. This is more common in countries with average income than in high-income countries [ 17 ]. Compared to Western populations, exceptional sensitivity in a particular population caused by a particular gene can have a strong impact on the clinical development of the disease [ 18 ]. 3.2. Immunity of atherosclerosis Immunity to atherosclerosis depends on several factors including genetics, autoimmune disorders [ 19 ], damaged cholesterol accumulation [ 20 , 21 ] and inflammation caused by intestinal microorganisms [ 22 – 24 ]. For prevention and management, the introduction of a balanced diet, physical exercise and a healthy lifestyle to avoid smoking is important [ 21 – 24 ]. In recent years, many researchers have been studying inflammatory cells in AS. Macrophages can ingest lipids and form foam cells. It is also considered a major cell that plays an important role in the AS process [ 25 ]. Several studies have shown that T-cell types and numbers have significant changes in the initial stage. This suggests that the immune response occurs before plate instability and is associated with plate instability [ 25 , 26 ]. Abnormal expression of membership molecules and chemokines leads to recruitment of inflammatory cells at the site of the lesion. This is an important symptom of this stage [ 27 , 28 ]. Excessive deposits of low-density lipoprotein (LDL) within arteries can damage the vascular endothelium. LDL passes through cells in endothelial blood vessels (ECs) and oxidizes with oxidized lipoproteins to low density (OX-LDL) [ 28 ]. This causes cytokine release and recruitment of monocytes at the site of destruction, and initiates recruitment of polymorphone neutrophils. After absorption, neutrophils using T cells release monocytes differentiated into granulocytes and macrophages [ 29 ]. However, various components of the immune system play a contradictory role in the progression of atherosclerosis (Fig. 2 ). 4. Macrophage recruitment Violation of endothelial function of blood vessels is the first stage of AS. At this stage, vasomotor function is incompatible, vascular endothelial permeability increases, and local inflammatory responses are accompanied by a focal inflammatory response [ 30 ]. On the other hand, damaged endothelial cells secrete monocytes associated with molecular (wet) models and damage (MCP-1) [ 31 ]. Adenosine Triphosphate (ATP), a key compound released from the death of damaged endothelial cells, activates inflammation of the nod-, LRR and pirin domains containing local macrophage protein 3 (NLRP3), and trankers, proximal to caspase-1 (IL-1). This ultimately leads to the expression of intercellular adhesion molecules (ICAMs) on endothelial cells that help adhesion of macrophages [ 32 ]. Macrophages are divided into MI and M2 types. Lymphocytes play an important role in the development of AS. During the inflammatory process, IL-12 and IIL-18 released by pathological cells can induce the expression of transcription factor 1 specific for specific transcription factors (T-BET) in naive CD4 + T cells (Figs. 3 ). At the same time, it induces Th1 cell differentiation and interferon (IFN) production [ 33 ]. The proinflammatory effects of Th1 cells may contribute to training as a plate. At the same time, V cells can also distinguish between proinflammatory factors such as TNF-α. This also contributes to formation. [ 34 ]. M2 type M2 macrophages are induced and differentiated by TH2 IL-4, IL-13 cytokines, and others. Studies also show that the effect of recruiting PMNs on recruitment of mononuclear macrophages is extremely important in early training of AS [ 35 , 36 ]. 4.1. Changes in the phenotype of macrophages Because the metabolic state of macrophages determines functional properties, changes in the macrophage phenotypic (M1 to M2 phenotype) can be obtained using metabolic reprogramming [ 37 ]. M2-type macrophages rely heavily on the mitochondrial oxidative phosphorylation system (OXPHOS) [ 38 ], whereas M1-type macrophages are affected by anaerobic glycoly [ 39 , 40 ]. Thanks to expanded single-cell and multicellular technologies, scientists can identify various immune or non-immune cells in the atherosclerotic plate and analyze the function of various cellular submarines and their interaction networks [ 41 – 43 ]. These techniques have helped to identify different populations of atherosclerotic plates, including different types of T cells and macrophages with specific markers and functions. Analysis of atherosclerotic plates reveals new levels of cell heterogeneity and indicates that the plate contains both proinflammatory and anti-inflammatory populations of macrophages. Scientists can analyze interactions between different types of cells within the plaque to understand how intercellular communication stimulates plate inflammation and progression [ 44 ]. 4.2. Single-cell technology for Atherosclerosis Single-cell technology revolutionized atherosclerosis, allowing for unprecedented resolution of cellular and molecular composition of plates, and detecting cellular heterogeneity, plasticity, and cell-to-cell connections in plate microism [ 45 ]. Methods such as sequencing of unicellular RNA (SCRNA-EQ) identify various sub-brasses of immune cells and functional status discovered by new therapeutic targets, and consider disease and risk factors progression [ 46 ]. These techniques play an important role in increasing accurate drugs and provide a cellular understanding required for the development of individual immunotherapies for atherosclerosis [ 46 ]. Over the past two years, single-cell technology has significantly increased our knowledge about atherosclerosis. SCRNA-EQ has been specifically used to characterize the landscape of murine immune cells and human atherosclerotic lesions [ 45 – 49 ]. Recent research by Fernandez et al . He provided the first complex overview of the landscape of human immune cells during atherosclerosis and presented a set of data based on time-of-flight analysis and extensive race motivational measures with comparisons of RNA expression profiles and macrophages in plates and blood of symptomatic and asymptomatic patients [ 49 ]. They give the idea of ​​immune cells on the plate and explain the different activation states. However, in both mouse studies and humans, there is not sufficient covering of non-immune cells in the plate. So far, only a limited number of patients have been included in the SCRNA-EQ study. Based on the expression of RNA expression and access to individual cell chromatin, the presence of proinflammatory and cytotoxic populations of T cells, multiple states of macrophage activation, and interactions with functionally different atherosclerotic plates can be considered modulators of human disease. 4.3. Single-cell immunity of atherosclerotic plates Most studies in this field are based on the use of animal models that fail to properly mimic the human immune environment and do not experience spontaneous rupture of the plate [ 50 , 51 ]. As a result, most studies focused on macrophages as motors in plate plates [ 52 , 53 ]. Nevertheless, an important but ambiguous role for other immune cells in place of plates, which are inherent to the immunological diversity of atherosclerotic plates, is expected. For example, mouse studies suggest T cells obtained from plates, but the phenotypic and functional diversity of these cells and other types of atherosclerosis and contributions to their illnesses continue to be found. Several studies have shown that circulating immune cells affect the clinical evolution of atherosclerosis, as patients with acute cardiovascular events exhibit monocytes and CD4 + circulating subassemblies [ 54 – 59 ]. 4.4. Macrophages of atherosclerotic plates In atherosclerosis, understanding of macrophages has evolved from populations to single cell biology, while being described by a subset of macrophages. It is important to note that the identity and functional specialization of macrophages can be very dynamic and depends on the fabric and molecular context [ 60 ]. In addition to recent changes in the paradigm in macrophage biology regarding health and disease, opinions regarding macrophage phenotypes and atherosclerotic function have also changed significantly [ 61 ]. In particular, single-cell crenelation of RNA (ScrnaseQ) has made it possible to identify new mouse subsets in human atherosclerosis [ 62 – 65 ]. Although data on the various transcription of macrophages in atherosclerosis increases, the effects of molecular and cellular contexts on phenotype and macrophage function have not yet been studied. However, the emergence of technological achievements such as multiple visualization and spatial transcription can help fill this gap by identifying macrophage phenotypes in spatial contexts [ 66 , 67 ]. 4.5. Origin of macrophages in atherosclerotic plates. Atherosclerotic plate macrophages mainly come from several sources that circulate from myelomonocytes that penetrate the arterial walls and proliferate macrophages and transfusion cells present in vascular smooth muscle (VSMC). Macrophages obtained from yellow also contribute to the early stages of birth, especially at the postnatal stage. These different origins lead to heterogeneous populations of macrophages within the plate. Each has a unique feature that affects plate progression, stability, and potential regression [ 68 ]. Initially and in progression of atherosclerosis, macrophages on the atherosclerotic plate vary in origin. Proliferation of residential macrophages in the atherosclerotic plate is not sufficient for long-term maintenance of glowing cells, and is largely fueled by supplemented monocytes, which play an important role in the expansion of macrophage populations during atherosclerosis [ 68 ]. Härdtner and his colleagues have shown that lipid reduction strategies inhibit local proliferation of macrophages in APOE*3-Leiden mice, indicating a decrease in the number of macrophages and plaque resolutions. Overall, this implies the importance of residents and the local reproduction of macrophages within the plate, rather than monocyte filling [ 69 ]. Furthermore, it has been shown that 40–60% of macrophages in plates have mezensimachus origin [ 70 ]. 4.6. Heterologous macrophages in the atherosclerotic plate. Microangles in which macrophages are phenotypic and functional specialization. Particularly in atherosclerotic state, this microflexion is significantly modified, highly complex, very complex, due to a wide range of phenotypes and macrophages, instrumental/deterministic, and ultimately stimulates plate progression and determines plate stabilization. Zernak et al . He demonstrated that despite differences in the rich studies of subassembly, it is possible to identify five major phenotypes of plate macrophages according to transcriptional profiles. It should be noted that various macrophages increase with progression of atherosclerosis [ 71 ]. The introduction of SCRNaseQ in the field of cardiovascular research has contributed significantly to the discovery and determination of macrophage subsets of atherosclerotic plates. Nevertheless, SCRNASEQ data, which played an important role in macrophage identification, may skip a specific subset and have no information about the spatial context. Therefore, it is extremely important to use methods that preserve tissue spatial composition to understand how biochemical and cellular regions affect phenotype and macrophage function. The different subsets of macrophages, their location, their functional aspects, and their origin are determined by comparing multipoint visualizations of advanced atherosclerotic plaques in mice at the root of the aorta. The colored background standing behind the subset symbolizes the segmentation of mouse atherosclerotic macrophages based on context [ 72 ]. 4.7. Macrophage Subset in Human Atherosclerosis Several sets of single-cell and mass cytometry identified a new subset of macrophages with human atherosclerosis [ 62 – 65 ]. The resident subset of macrophages is characterized by the expression of genes such as Lyve1, CD206, CD163, and FRANR2 [ 73 ]. Mass Citometry divided them into two resident subassemblies: CD206HI CD163HI and CD206LO CD163LO are present in symptomatic and asymptomatic plates [ 63 ]. Previous studies have linked CD163 + related macrophages with a more advanced plate phenotype cooked in plates, with a marker of plates and thrombotic events [ 74 , 75 ]. CD163-expressing macrophages are present in the shoulder area of ​​the plate [ 75 ], and the main CD68-expressing macrophages were found in addiction [ 74 ]. It is interesting to note that the role of macrophage CD163 + in atherosclerosis varies from proinflammatory [ 76 ] to anti-inflammatory [ 75 ]. The association with different components of different locations, plates [ 74 , 75 ] and a subset of macrophages of expressed CD163 is a possible explanation of functional divergence transmitted due to atherosclerosis [ 63 ]. M1 macrophages produce low levels of high levels of inflammatory cytokines, particularly IL-12, IL-23, IL-6, IL-1β, IL-8, TNF-α, and the anti-inflammatory cytokine IL-10. Furthermore (Figs. 4 ), they exhibit increased microbial agent activity, releasing high levels of active forms of oxygen and nitrogen radicals [ 77 ]. Unlike M1, a proinflammatory macrophage, the M2 phenotype shows high levels of transformation growth factors (TGF)-β and IL-10, and low levels of IL-12 and IL-23. M2 macrophages express high levels of mannose receivers (CD206) and contribute to wound healing using germinatria, matrix reinvasion, and fibroblast recruitment [ 78 ]. In addition to TH, the complex microenvironment of atherosclerotic lesions induces the phenotypic differentiation of resident macrophages, including macrophages that respond to oxidized phenotypes such as hemoglobin (MB), macrophages, and M4 M4 macrophages. Various subpopulations of macrophages associated with the presence of hemoglobin and red blood cells have been identified in the hemorrhagic region of the human atherosclerotic plate [ 79 ]. 5. Conclusion Although the location, heterogeneity, and the function of human atherosclerosis have been rarely studied, lipid-related macrophages with similar expression of genes in other disease and hemostasis contexts have been identified in mice and people. Ultimately, macrophages should provide the benefits or benefits of a niche. In atherosclerosis, the benefits of the niche may include macrophages such as dummy clearance of lipid and cell, stimulating angiogenesis, increased fibrosis, and stabilizing fibrotic caps. The concept of macrophage niches with atherosclerosis has not yet been studied, but plates vary in biochemical and mechanical microinflation. Future research should use in situ methods such as multipoint visualization and spatial transcriptomes to obtain a deeper idea of ​​human atherosclerotic bubble submime and its richness and corresponding functions. Nevertheless, different levels of macrophages can express cell surfaces that determine different levels of transcriptional markers and subsets. It is not yet clear how these different expression profiles reflect differences in niche, location, and functional composition. Therefore, more complete properties of the macrophage subbras in the corresponding plate frame are required. Such properties should be based on the complete profile of transcriptional markers and/or surfaces combined with information on histological placement of the subset, cellular and molecular textures, and connections with plate properties. The rich properties of atherosclerotic macrophage niches and linkage with useful and harmful macrophage wells help develop therapeutic approaches aimed at targeting sections or macrophages by modifying the properties in microsacrons. Declarations Funding No Funding. Conflict of Interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Human and Animal Rights and Informed Consent No animal or human subjects by the authors were used in this study. Consent for Publication All authors reviewed the results and approved the final version of the manuscript. Authors’ Contributions Hossein Nasouti Aghjehroud, Hamed Aghazadeh and Sobhan Aboulhassanzadeh: writing manuscript; Roghayeh Alizadeh, Aryan Mahmood Faraj, Hamed Aghazadeh and Samin Aboulhassanzadeh: conceptualization, data curation, investigation, methodology, validation, visualization, writing-original draft. All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work. Availability of Data and Materials All relevant data can be found within the manuscript. Acknowledgement The authors would like to present their gratitude to the Dublin City University, Dublin, Ireland for supporting this study. Key References • Cetin E & Raby AC. Understanding atherosclerotic plaque cellular composition: recent advances driven by single cell omics. 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Targeting diabetic atherosclerosis: the role of GLP-1 receptor agonists, SGLT2 inhibitors, and nonsteroidal mineralocorticoid receptor antagonists in vascular protection and disease modulation. Biomedicines. 2025;13(3), 728. ‏ https://doi.org/10.3390/biomedicines13030728 Park, M. D., Silvin, A., Ginhoux, F. and et al. Macrophages in health and disease. Cell. 2022; 185: 4259–4279. https://doi.org/10.1016/j.cell.2022.10.007 Zernecke A, Winkels H, Cochain C and et al. Meta-Analysis of leukocyte diversity in atherosclerotic mouse aortas. Circ Res. 2020;127: 402–426. https://doi.org/10.1161/CIRCRESAHA.120.316903 Winkels H, Ehinger E, Vassallo M and et al. Atlas of the immune cell repertoire in mouse atherosclerosis defined by Single-Cell RNA-Sequencing and mass cytometry. Circ Res. 2018;122: 1675–1688. https://doi.org/10.1161/CIRCRESAHA.117.312513 Fernandez DM, Rahman AH, Fernandez NF and et al. Single-cell immune landscape of human atherosclerotic plaques. Nat Med. 2019;25: 1576–1588. https://doi.org/10.1038/s41591-019-0590-4 Wirka RC, Wagh D, Paik DT and et al. Atheroprotective roles of smooth muscle cell phenotypic modulation and the TCF21 disease gene as revealed by single-cell analysis . Nat Med. 2019;25: 1280–1289. https://doi.org/10.1038/s41591-019-0512-5 Depuydt MAC, Prange KHM, Slenders L and et al. Microanatomy of the human atherosclerotic plaque by Single-Cell transcriptomics. Circ Res. 2020;127: 1437–1455. https://doi.org/10.1161/CIRCRESAHA.120.316770 Goossens P, Lu C, Cao J and et al. Integrating multiplex immunofluorescent and mass spectrometry imaging to map myeloid heterogeneity in its metabolic and cellular context. Cell Metab. 2022;34: 1214–1225 e6. https://doi.org/10.1016/j.cmet.2022.06.012 Marx V. Method of the year 2020: spatially resolved transcriptomics. Nat Methods . 2021;18: 1. https://doi.org/10.1038/s41592-020-01033-y Williams JW, Zaitsev K, Kim KW and et al. Limited proliferation capacity of aortic intima resident macrophages requires monocyte recruitment for atherosclerotic plaque progression. Nat Immunol . 2020;21: 1194–1204. https://doi.org/10.1038/s41590-020-0768-4 Hardtner C, Kornemann J, Krebs K and et al. Inhibition of macrophage proliferation dominates plaque regression in response to cholesterol lowering. Basic Res Cardiol. 2020;115: 78. https://doi.org/10.1007/s00395-020-00838-4 Sadykova D, Nigmatullina R, Salakhova K and et al. Role of Serotonin, Membrane Transporter, and 5-HT2 Receptors in Pathogenesis of Atherosclerotic Plaque Formation in Immature Heterozygous Low-Density Lipoprotein-Receptor-Deficient Mice. Int J Mol Sci. 2025;26(13), 6184. ‏ https://doi.org/10.3390/ijms26136184 Zernecke A, Winkels H, Cochain C and et al. Meta-Analysis of leukocyte diversity in atherosclerotic mouse aortas. Circ Res . 2020;127: 402–426. https://doi.org/10.1161/CIRCRESAHA.120.316903 Goossens P, Lu C, Cao J and et al. Integrating multiplex immunofluorescent and mass spectrometry imaging to map myeloid heterogeneity in its metabolic and cellular context. Cell Metab. 2022;34: 1214–1225 e6. https://doi.org/10.1016/j.cmet.2022.06.012 Zernecke A, Erhard F, Weinberger T and et al. Integrated single-cell analysis-based classification of vascular mononuclear phagocytes in mouse and human atherosclerosis. Cardiovasc Res. 2022;119: 1676–1689. https://doi.org/10.1093/cvr/cvac161 Stoger JL, Gijbels MJ, Van S and et al. Distribution of macrophage polarization markers in human atherosclerosis. Atherosclerosis. 2012;225: 461–468. https://doi.org/10.1016/j.atherosclerosis.2012.09.013 Bengtsson E, Hultman K, Edsfeldt A and et al. CD163+ macrophages are associated with a vulnerable plaque phenotype in human carotid plaques. Sci Rep. 2020;10: 14362. https://doi.org/10.1038/s41598-020-71110-x Guo L, Akahori H, Harari E and et al. CD163+ macrophages promote angiogenesis and vascular permeability accompanied by inflammation in atherosclerosis. J Clin Invest. 2018;128: 1106–1124. https://doi.org/10.1172/JCI93025 Cutolo M, Soldano S, Smith V and et al. Dynamic macrophage phenotypes in autoimmune and inflammatory rheumatic diseases. Nat Rev Rheumatol. 2025;1-20. ‏ https://doi.org/10.1038/s41584-025-01279-w Peng M, Zhu Y, Hu Y and et al. Advances in the regulation of macrophage polarization by the tumor microenvironment. Discov Oncol. 2025;16(1), 1487. ‏ https://doi.org/10.1007/s12672-025-03258-9 KANDE R, PASHA A, KANTIPUDI S and et al. HAPTOGLOBIN: AN ACUTE PHASE RESPONSE PROTEIN WITH DIVERSE ROLES IN INFECTION AND SEPSIS. AJMBES. 2025;27 (3-4): 151-162. http://doi.org/10.53550/AJMBES.2025.v27i03-04.004 Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7393034","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Systematic Review","associatedPublications":[],"authors":[{"id":589740320,"identity":"d2c5ffd8-9d6d-476c-8eb7-4a0b446f2955","order_by":0,"name":"Hossein Nasouti Aghjehroud","email":"","orcid":"","institution":"University of Turin","correspondingAuthor":false,"prefix":"","firstName":"Hossein","middleName":"Nasouti","lastName":"Aghjehroud","suffix":""},{"id":589740321,"identity":"d5ec0a71-e5fa-4b5e-914f-bd8c2d3ee6d6","order_by":1,"name":"Roghayeh Alizadeh","email":"","orcid":"","institution":"Pasteur Institute of Iran","correspondingAuthor":false,"prefix":"","firstName":"Roghayeh","middleName":"","lastName":"Alizadeh","suffix":""},{"id":589740322,"identity":"229d1883-031b-4db1-828b-3e7fe1ff3157","order_by":2,"name":"Aryan Mahmood Faraj","email":"","orcid":"","institution":"Halabja Technical College, Sulaimani Polytechnic University","correspondingAuthor":false,"prefix":"","firstName":"Aryan","middleName":"Mahmood","lastName":"Faraj","suffix":""},{"id":589740323,"identity":"c8322d58-8cee-4c7d-ad32-793fd53abf87","order_by":3,"name":"Hamed Aghazadeh","email":"","orcid":"","institution":"Tabriz University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Hamed","middleName":"","lastName":"Aghazadeh","suffix":""},{"id":589740324,"identity":"3249fa4b-ac02-472c-a079-f7059fd979b9","order_by":4,"name":"Samin Aboulhassanzadeh","email":"","orcid":"","institution":"Mérieux NutriSciences - Global","correspondingAuthor":false,"prefix":"","firstName":"Samin","middleName":"","lastName":"Aboulhassanzadeh","suffix":""},{"id":589740325,"identity":"0e84ad73-4f8b-4ae9-b60b-7a2c0ede109b","order_by":5,"name":"Sobhan Aboulhassanzadeh","email":"data:image/png;base64,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","orcid":"","institution":"Dublin City University","correspondingAuthor":true,"prefix":"","firstName":"Sobhan","middleName":"","lastName":"Aboulhassanzadeh","suffix":""}],"badges":[],"createdAt":"2025-08-17 14:53:08","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7393034/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7393034/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":102496795,"identity":"8155a821-532f-4651-bae8-50506ac71f47","added_by":"auto","created_at":"2026-02-12 09:44:35","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":426179,"visible":true,"origin":"","legend":"\u003cp\u003eProgression of Atherosclerosis\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7393034/v1/a65134ddcd8f1020f33178a3.png"},{"id":102746353,"identity":"928494ae-f0d8-4efd-9629-60f099dd7e06","added_by":"auto","created_at":"2026-02-16 08:56:57","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1725174,"visible":true,"origin":"","legend":"\u003cp\u003eDifferent components of the immune system in relation to the progression of atherosclerosis\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7393034/v1/4e68696fd746644f9be3d43e.png"},{"id":102746815,"identity":"e0aed109-8c26-470f-a58b-21c69ef110c1","added_by":"auto","created_at":"2026-02-16 09:01:35","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":671245,"visible":true,"origin":"","legend":"\u003cp\u003eMacrophage recruitment\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7393034/v1/fe0212420bae546bf44fb9b6.png"},{"id":102496799,"identity":"46439b81-7d0d-442f-b206-ee527c550914","added_by":"auto","created_at":"2026-02-12 09:44:39","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":486188,"visible":true,"origin":"","legend":"\u003cp\u003eMacrophage phenotypes in atherosclerotic plaque. The different microenvironments present in atherosclerotic plaque drive\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7393034/v1/b1824f83ee1438d5d331aca1.png"},{"id":102750619,"identity":"40e6c26e-a01f-4875-be9f-951046411bc3","added_by":"auto","created_at":"2026-02-16 09:20:51","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4124809,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7393034/v1/4c847fd0-db86-4b6f-a2b8-912546fee397.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"A Review of Macrophage Diversity and Immune Checkpoint Regulation in Atherosclerosis: Insights from Single-Cell Transcriptomes","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eAtherosclerosis (AS) is a complex metabolic and inflammatory disease of the arterial wall, characterized by the accumulation of lipoproteins rich in cholesterol and inflammatory cells. Accumulation of lipoproteins containing lipoproteins containing apolipoprotein B Atherosclerosis is the main cause of cardiovascular events such as myocardial infarction and stroke, resulting in 17.9\u0026nbsp;million deaths worldwide [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. The progression of the atherosclerotic plate is preferred due to hypercholesterolemia and dyslipidemia. Its development begins when macrophages living in the inner membrane occupy excess lipids and form bubble cells. Over the past 20 years, achievements in the field of atherosclerosis have identified myeloidemia, the proliferation of plaque and steatosis macrophages as an important determinant of atherosclerosis development and regression. These parameters have recently been shown to be affected by immune cell metabolism [\u003cspan additionalcitationids=\"CR3\" citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. It is interesting to note that while the clearance of lipoproteins by macrophages plays a similar role in preventing the production of harmful lipids, excessive lipid metabolism in macrophages leads to phenotypic and function changes, ultimately leading to death [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Macrophages, essential immune cells, are truly known for their remarkable ability to accept a variety of phenotypes and functions depending on the environment, and recently, sequencing of single-cell RNA RNAs has significantly increased our understanding of macrophage heterogeneity in atherosclerotic zone classification [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThis technique provides a more detailed description of these cells and their role in the potential progression and treatment of atherosclerosis. SCRNA-EQ provides molecular profiles to individual macrophages with high resolution, allowing researchers to identify unique gene expression models and derive specific functions of various water macrophages in atherosclerotic lesions [\u003cspan additionalcitationids=\"CR8\" citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAs a result, understanding the phenotypic complexity of macrophages is important for developing effective therapeutic strategies. Aiming for modulation of a specific subset of macrophages or their function satisfies a perspective to prevent or modify plate progression. Because macrophages play an important role in all stages of atherosclerosis and are involved in inflammation, lipid accumulation and plate evolution are important for different subpopulations to develop targeted therapies. As a result, this study aims to discuss the regulation of various macrophages and immune control points in atherosclerosis.\u003c/p\u003e \u003cp\u003eThis discussion overturned unprecedented light on the transcriptional environment and phenotypic heterogeneity of atherosclerotic macrophages, exhibited special functions, and opened up new opportunities to study different populations and their functions in different populations of atherosclerosis.\u003c/p\u003e"},{"header":"2. Research Methodology","content":"\u003cp\u003eThe design of the study was a review. This study looked for and investigated articles related to various macrophages of atherosclerosis. Particularly, we understood the base using PubMed, Web of Science (clariivate Analytics), Scopus, and Kokran\u0026eacute;e libraries and the appropriate keywords, and were associated with a variety of macrophages in atherosclerosis. The study was conducted in June 2025 and approved by the counterpart council. We are also considering Google Scholar to find the right item. It also includes a list of article links.\u003c/p\u003e"},{"header":"3. Progression of atherosclerosis","content":"\u003cp\u003eAtherosclerosis has been increasing over the years through various stages of endothelial damage, the formation of fat bands, plate growth, rupture of plates or erosion, causing potential complications such as heart attacks and strokes [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. This process is universal, but accelerates risk factors such as high cholesterol, high blood pressure, diabetes, smoking and obesity (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). This process begins with damage to the internal mucosa of the artery (endothelium). This causes an immune response in which the white blood is replenished in the field. LDL cholesterol (bad cholesterol) accumulates and is associated with damaged endothelium, forming early deposits called fatty stripes [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. These fat bands develop in more complex plates consisting of cholesterol, fatty substances, calcium and cell waste. Fibrous coverage is formed on the plate and can be stabilized or unstable [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. The plate can explode, which leads to the formation of a blood clot (thrombosis). This thrombus can seriously or completely block the artery and, if it affects the brain, affecting the heart or stroke, causes a heart attack [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. On the other hand, macrophages rapidly recognize and collect LDL-C via surface binge, and convert them into cells to form the first atherosclerotic lesions [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. However, it becomes clear that lipid reduction alone does not completely interfere with the progression of atherosclerosis, revealing that macrophages are new potential therapeutic targets.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Epidemiology of atherosclerosis\u003c/h2\u003e \u003cp\u003eAtherosclerosis primarily affects the heart and brain, manifesting as coronary artery disease and ischemic stroke, and according to a 2019 epidemiological study, there are places that cannot be denied in the choice of mortality around the world [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. The World Health Organization has shown that coronary arteries and stroke together account for around 85% of deaths, exceeding other causes, including cancer [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. This is more common in countries with average income than in high-income countries [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Compared to Western populations, exceptional sensitivity in a particular population caused by a particular gene can have a strong impact on the clinical development of the disease [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Immunity of atherosclerosis\u003c/h2\u003e \u003cp\u003eImmunity to atherosclerosis depends on several factors including genetics, autoimmune disorders [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e], damaged cholesterol accumulation [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e] and inflammation caused by intestinal microorganisms [\u003cspan additionalcitationids=\"CR23\" citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. For prevention and management, the introduction of a balanced diet, physical exercise and a healthy lifestyle to avoid smoking is important [\u003cspan additionalcitationids=\"CR22 CR23\" citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. In recent years, many researchers have been studying inflammatory cells in AS. Macrophages can ingest lipids and form foam cells. It is also considered a major cell that plays an important role in the AS process [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Several studies have shown that T-cell types and numbers have significant changes in the initial stage. This suggests that the immune response occurs before plate instability and is associated with plate instability [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Abnormal expression of membership molecules and chemokines leads to recruitment of inflammatory cells at the site of the lesion. This is an important symptom of this stage [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Excessive deposits of low-density lipoprotein (LDL) within arteries can damage the vascular endothelium. LDL passes through cells in endothelial blood vessels (ECs) and oxidizes with oxidized lipoproteins to low density (OX-LDL) [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. This causes cytokine release and recruitment of monocytes at the site of destruction, and initiates recruitment of polymorphone neutrophils. After absorption, neutrophils using T cells release monocytes differentiated into granulocytes and macrophages [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. However, various components of the immune system play a contradictory role in the progression of atherosclerosis (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4. Macrophage recruitment","content":"\u003cp\u003eViolation of endothelial function of blood vessels is the first stage of AS. At this stage, vasomotor function is incompatible, vascular endothelial permeability increases, and local inflammatory responses are accompanied by a focal inflammatory response [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. On the other hand, damaged endothelial cells secrete monocytes associated with molecular (wet) models and damage (MCP-1) [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Adenosine Triphosphate (ATP), a key compound released from the death of damaged endothelial cells, activates inflammation of the nod-, LRR and pirin domains containing local macrophage protein 3 (NLRP3), and trankers, proximal to caspase-1 (IL-1). This ultimately leads to the expression of intercellular adhesion molecules (ICAMs) on endothelial cells that help adhesion of macrophages [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Macrophages are divided into MI and M2 types. Lymphocytes play an important role in the development of AS. During the inflammatory process, IL-12 and IIL-18 released by pathological cells can induce the expression of transcription factor 1 specific for specific transcription factors (T-BET) in naive CD4\u0026thinsp;+\u0026thinsp;T cells (Figs.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). At the same time, it induces Th1 cell differentiation and interferon (IFN) production [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. The proinflammatory effects of Th1 cells may contribute to training as a plate. At the same time, V cells can also distinguish between proinflammatory factors such as TNF-α. This also contributes to formation. [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. M2 type M2 macrophages are induced and differentiated by TH2 IL-4, IL-13 cytokines, and others. Studies also show that the effect of recruiting PMNs on recruitment of mononuclear macrophages is extremely important in early training of AS [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e4.1. Changes in the phenotype of macrophages\u003c/h2\u003e \u003cp\u003eBecause the metabolic state of macrophages determines functional properties, changes in the macrophage phenotypic (M1 to M2 phenotype) can be obtained using metabolic reprogramming [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. M2-type macrophages rely heavily on the mitochondrial oxidative phosphorylation system (OXPHOS) [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e], whereas M1-type macrophages are affected by anaerobic glycoly [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. Thanks to expanded single-cell and multicellular technologies, scientists can identify various immune or non-immune cells in the atherosclerotic plate and analyze the function of various cellular submarines and their interaction networks [\u003cspan additionalcitationids=\"CR42\" citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. These techniques have helped to identify different populations of atherosclerotic plates, including different types of T cells and macrophages with specific markers and functions. Analysis of atherosclerotic plates reveals new levels of cell heterogeneity and indicates that the plate contains both proinflammatory and anti-inflammatory populations of macrophages. Scientists can analyze interactions between different types of cells within the plaque to understand how intercellular communication stimulates plate inflammation and progression [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e4.2. Single-cell technology for Atherosclerosis\u003c/h2\u003e \u003cp\u003eSingle-cell technology revolutionized atherosclerosis, allowing for unprecedented resolution of cellular and molecular composition of plates, and detecting cellular heterogeneity, plasticity, and cell-to-cell connections in plate microism [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. Methods such as sequencing of unicellular RNA (SCRNA-EQ) identify various sub-brasses of immune cells and functional status discovered by new therapeutic targets, and consider disease and risk factors progression [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. These techniques play an important role in increasing accurate drugs and provide a cellular understanding required for the development of individual immunotherapies for atherosclerosis [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. Over the past two years, single-cell technology has significantly increased our knowledge about atherosclerosis. SCRNA-EQ has been specifically used to characterize the landscape of murine immune cells and human atherosclerotic lesions [\u003cspan additionalcitationids=\"CR46 CR47 CR48\" citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. Recent research by Fernandez \u003cem\u003eet al\u003c/em\u003e. He provided the first complex overview of the landscape of human immune cells during atherosclerosis and presented a set of data based on time-of-flight analysis and extensive race motivational measures with comparisons of RNA expression profiles and macrophages in plates and blood of symptomatic and asymptomatic patients [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. They give the idea of ​​immune cells on the plate and explain the different activation states. However, in both mouse studies and humans, there is not sufficient covering of non-immune cells in the plate. So far, only a limited number of patients have been included in the SCRNA-EQ study. Based on the expression of RNA expression and access to individual cell chromatin, the presence of proinflammatory and cytotoxic populations of T cells, multiple states of macrophage activation, and interactions with functionally different atherosclerotic plates can be considered modulators of human disease.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e4.3. Single-cell immunity of atherosclerotic plates\u003c/h2\u003e \u003cp\u003eMost studies in this field are based on the use of animal models that fail to properly mimic the human immune environment and do not experience spontaneous rupture of the plate [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e, \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. As a result, most studies focused on macrophages as motors in plate plates [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e]. Nevertheless, an important but ambiguous role for other immune cells in place of plates, which are inherent to the immunological diversity of atherosclerotic plates, is expected. For example, mouse studies suggest T cells obtained from plates, but the phenotypic and functional diversity of these cells and other types of atherosclerosis and contributions to their illnesses continue to be found. Several studies have shown that circulating immune cells affect the clinical evolution of atherosclerosis, as patients with acute cardiovascular events exhibit monocytes and CD4\u0026thinsp;+\u0026thinsp;circulating subassemblies [\u003cspan additionalcitationids=\"CR55 CR56 CR57 CR58\" citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e4.4. Macrophages of atherosclerotic plates\u003c/h2\u003e \u003cp\u003eIn atherosclerosis, understanding of macrophages has evolved from populations to single cell biology, while being described by a subset of macrophages. It is important to note that the identity and functional specialization of macrophages can be very dynamic and depends on the fabric and molecular context [\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]. In addition to recent changes in the paradigm in macrophage biology regarding health and disease, opinions regarding macrophage phenotypes and atherosclerotic function have also changed significantly [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e]. In particular, single-cell crenelation of RNA (ScrnaseQ) has made it possible to identify new mouse subsets in human atherosclerosis [\u003cspan additionalcitationids=\"CR63 CR64\" citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e]. Although data on the various transcription of macrophages in atherosclerosis increases, the effects of molecular and cellular contexts on phenotype and macrophage function have not yet been studied. However, the emergence of technological achievements such as multiple visualization and spatial transcription can help fill this gap by identifying macrophage phenotypes in spatial contexts [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e, \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e4.5. Origin of macrophages in atherosclerotic plates.\u003c/h2\u003e \u003cp\u003eAtherosclerotic plate macrophages mainly come from several sources that circulate from myelomonocytes that penetrate the arterial walls and proliferate macrophages and transfusion cells present in vascular smooth muscle (VSMC). Macrophages obtained from yellow also contribute to the early stages of birth, especially at the postnatal stage. These different origins lead to heterogeneous populations of macrophages within the plate. Each has a unique feature that affects plate progression, stability, and potential regression [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. Initially and in progression of atherosclerosis, macrophages on the atherosclerotic plate vary in origin. Proliferation of residential macrophages in the atherosclerotic plate is not sufficient for long-term maintenance of glowing cells, and is largely fueled by supplemented monocytes, which play an important role in the expansion of macrophage populations during atherosclerosis [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. H\u0026auml;rdtner and his colleagues have shown that lipid reduction strategies inhibit local proliferation of macrophages in APOE*3-Leiden mice, indicating a decrease in the number of macrophages and plaque resolutions. Overall, this implies the importance of residents and the local reproduction of macrophages within the plate, rather than monocyte filling [\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e]. Furthermore, it has been shown that 40\u0026ndash;60% of macrophages in plates have mezensimachus origin [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e4.6. Heterologous macrophages in the atherosclerotic plate.\u003c/h2\u003e \u003cp\u003eMicroangles in which macrophages are phenotypic and functional specialization. Particularly in atherosclerotic state, this microflexion is significantly modified, highly complex, very complex, due to a wide range of phenotypes and macrophages, instrumental/deterministic, and ultimately stimulates plate progression and determines plate stabilization. Zernak \u003cem\u003eet al\u003c/em\u003e. He demonstrated that despite differences in the rich studies of subassembly, it is possible to identify five major phenotypes of plate macrophages according to transcriptional profiles. It should be noted that various macrophages increase with progression of atherosclerosis [\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e]. The introduction of SCRNaseQ in the field of cardiovascular research has contributed significantly to the discovery and determination of macrophage subsets of atherosclerotic plates. Nevertheless, SCRNASEQ data, which played an important role in macrophage identification, may skip a specific subset and have no information about the spatial context. Therefore, it is extremely important to use methods that preserve tissue spatial composition to understand how biochemical and cellular regions affect phenotype and macrophage function. The different subsets of macrophages, their location, their functional aspects, and their origin are determined by comparing multipoint visualizations of advanced atherosclerotic plaques in mice at the root of the aorta. The colored background standing behind the subset symbolizes the segmentation of mouse atherosclerotic macrophages based on context [\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e4.7. Macrophage Subset in Human Atherosclerosis\u003c/h2\u003e \u003cp\u003eSeveral sets of single-cell and mass cytometry identified a new subset of macrophages with human atherosclerosis [\u003cspan additionalcitationids=\"CR63 CR64\" citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e]. The resident subset of macrophages is characterized by the expression of genes such as Lyve1, CD206, CD163, and FRANR2 [\u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\u003c/span\u003e]. Mass Citometry divided them into two resident subassemblies: CD206HI CD163HI and CD206LO CD163LO are present in symptomatic and asymptomatic plates [\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e]. Previous studies have linked CD163\u0026thinsp;+\u0026thinsp;related macrophages with a more advanced plate phenotype cooked in plates, with a marker of plates and thrombotic events [\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e, \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e]. CD163-expressing macrophages are present in the shoulder area of ​​the plate [\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e], and the main CD68-expressing macrophages were found in addiction [\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e]. It is interesting to note that the role of macrophage CD163\u0026thinsp;+\u0026thinsp;in atherosclerosis varies from proinflammatory [\u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e76\u003c/span\u003e] to anti-inflammatory [\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e]. The association with different components of different locations, plates [\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e, \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e] and a subset of macrophages of expressed CD163 is a possible explanation of functional divergence transmitted due to atherosclerosis [\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eM1 macrophages produce low levels of high levels of inflammatory cytokines, particularly IL-12, IL-23, IL-6, IL-1β, IL-8, TNF-α, and the anti-inflammatory cytokine IL-10. Furthermore (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e), they exhibit increased microbial agent activity, releasing high levels of active forms of oxygen and nitrogen radicals [\u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e77\u003c/span\u003e]. Unlike M1, a proinflammatory macrophage, the M2 phenotype shows high levels of transformation growth factors (TGF)-β and IL-10, and low levels of IL-12 and IL-23. M2 macrophages express high levels of mannose receivers (CD206) and contribute to wound healing using germinatria, matrix reinvasion, and fibroblast recruitment [\u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e78\u003c/span\u003e]. In addition to TH, the complex microenvironment of atherosclerotic lesions induces the phenotypic differentiation of resident macrophages, including macrophages that respond to oxidized phenotypes such as hemoglobin (MB), macrophages, and M4 M4 macrophages. Various subpopulations of macrophages associated with the presence of hemoglobin and red blood cells have been identified in the hemorrhagic region of the human atherosclerotic plate [\u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e79\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eAlthough the location, heterogeneity, and the function of human atherosclerosis have been rarely studied, lipid-related macrophages with similar expression of genes in other disease and hemostasis contexts have been identified in mice and people. Ultimately, macrophages should provide the benefits or benefits of a niche. In atherosclerosis, the benefits of the niche may include macrophages such as dummy clearance of lipid and cell, stimulating angiogenesis, increased fibrosis, and stabilizing fibrotic caps. The concept of macrophage niches with atherosclerosis has not yet been studied, but plates vary in biochemical and mechanical microinflation. Future research should use in situ methods such as multipoint visualization and spatial transcriptomes to obtain a deeper idea of ​​human atherosclerotic bubble submime and its richness and corresponding functions. Nevertheless, different levels of macrophages can express cell surfaces that determine different levels of transcriptional markers and subsets. It is not yet clear how these different expression profiles reflect differences in niche, location, and functional composition. Therefore, more complete properties of the macrophage subbras in the corresponding plate frame are required. Such properties should be based on the complete profile of transcriptional markers and/or surfaces combined with information on histological placement of the subset, cellular and molecular textures, and connections with plate properties. The rich properties of atherosclerotic macrophage niches and linkage with useful and harmful macrophage wells help develop therapeutic approaches aimed at targeting sections or macrophages by modifying the properties in microsacrons.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo Funding.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHuman and Animal Rights and Informed Consent\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo animal or human subjects by the authors were used in this study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for Publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors reviewed the results and approved the final version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHossein Nasouti Aghjehroud, Hamed Aghazadeh and Sobhan Aboulhassanzadeh: writing manuscript; Roghayeh Alizadeh, Aryan Mahmood Faraj, Hamed Aghazadeh and Samin Aboulhassanzadeh: conceptualization, data curation, investigation, methodology, validation, visualization, writing-original draft. All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of Data and Materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll relevant data can be found within the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to present their gratitude to the Dublin City University, Dublin, Ireland for supporting this study.\u003c/p\u003e"},{"header":"Key References","content":"\u003cp\u003e \u003cb\u003e\u0026bull;\u003c/b\u003e Cetin E \u0026amp; Raby AC. Understanding atherosclerotic plaque cellular composition: recent advances driven by single cell omics. Cells. 2025;14(11), 770.\u0026rlm; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/cells14110770\u003c/span\u003e\u003cspan address=\"10.3390/cells14110770\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eThis review provides an overview of the latest findings in the field obtained using single-cell technologies, with a focus on the major cell types present at the atherosclerotic plaque site, their suggested repartition in subsets, as well as their predicted function(s) within the complex processes that interplay to drive atherosclerotic disease.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003e\u0026bull;\u003c/b\u003e Cochain C, Vafadarnejad E, Arampatzi P and et al. Single-Cell RNA-Seq reveals the transcriptional landscape and heterogeneity of aortic macrophages in murine atherosclerosis. Circ Res. 2018;122: 1661\u0026ndash;1674. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1161/CIRCRESAHA.117.312509\u003c/span\u003e\u003cspan address=\"10.1161/CIRCRESAHA.117.312509\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eUnprecedentedly uncovered the transcriptional landscape and phenotypic heterogeneity of aortic macrophages and monocyte-derived dendritic cells in atherosclerotic and identified previously unrecognized macrophage populations and their gene expression signature, suggesting specialized functions.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003e\u0026bull;\u003c/b\u003e Engelen SE, Robinson AJB, Zurke YX and et al. Therapeutic strategies targeting inflammation and immunity in atherosclerosis: How to proceed? Nat Rev Cardiol. 2022;19: 522\u0026ndash;542. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41569-021-00668-4\u003c/span\u003e\u003cspan address=\"10.1038/s41569-021-00668-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eThe essential role of immunological components in the development and chronicity of atherosclerosis has been recognized as a clinical target, and proven studies demonstrate the benefits and challenges of targeting inflammation and the immune system in cardiovascular diseases.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003e\u0026bull;\u003c/b\u003e Ji J, Lindberg EL \u0026amp; Reichart D. Advancing Cardiovascular Medicine: Innovative Therapeutic Pathways with Single-Cell Technologies. Curr Treat Options Cardio Med. 2025;27(1), 48.\u0026rlm; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11936-025-01100-7\u003c/span\u003e\u003cspan address=\"10.1007/s11936-025-01100-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eIntegrating personalized transcriptions into medical routines will also facilitate more accurate and effective interventions in cardiovascular medicine.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003e\u0026bull;\u003c/b\u003e Lai Z, Kong D, Li Q and et al. Single-cell spatial transcriptomics of tertiary lymphoid organ-like structures in human atherosclerotic plaques. Nat Cardiovasc Res. 2025;4(5), 547\u0026ndash;566.\u0026rlm; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s44161-025-00639-9\u003c/span\u003e\u003cspan address=\"10.1038/s44161-025-00639-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eIt reveals the characteristics of PTLOs in human atherosclerosis, their cellular functions and clinical implications, offering avenues for understanding, diagnosing and treating this disease.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003e\u0026bull;\u003c/b\u003e Liu X, Guo JW, Lin XC and et al. Macrophage NFATc3 prevents foam cell formation and atherosclerosis: evidence and mechanisms. Eur Heart J. 2021;42(47): 4847\u0026ndash;61. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/eurheartj/ehab660\u003c/span\u003e\u003cspan address=\"10.1093/eurheartj/ehab660\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eMacrophages prevent the formation of foam cells and atherosclerosis, which may be a potential therapeutic target against atherosclerosis.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003e\u0026bull;\u003c/b\u003e Nissen SE, Wolski K, Cho L et al. Lipoprotein(a) levels in a global population with established atherosclerotic cardiovascular disease. Open Heart. 2022;9: e002060. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1136/openhrt-2022-002060\u003c/span\u003e\u003cspan address=\"10.1136/openhrt-2022-002060\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eThis study sought to characterise patterns of Lp(a) levels in a global ASCVD population and identify racial, ethnic, regional and gender differences.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003e\u0026bull;\u003c/b\u003e Roy P, Orecchioni M \u0026amp; Ley K. How the immune system shapes atherosclerosis: Roles of innate and adaptive immunity. Nat Rev Immunol. 2022;22: 251\u0026ndash;265. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41577-021-00584-1\u003c/span\u003e\u003cspan address=\"10.1038/s41577-021-00584-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eUnderstanding the activation pathways associated with cardiovascular disease and their regulation by microbial and metabolic factors will be crucial for the development of clinical interventions for atherosclerosis, including potential vaccination-based therapeutic strategies.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003e\u0026bull;\u003c/b\u003e Song B, Bie Y, Feng H and et al. Inflammatory factors driving atherosclerotic plaque progression new insights. 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Circ Res. 2018;122: 1675\u0026ndash;1688. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1161/CIRCRESAHA.117.312513\u003c/span\u003e\u003cspan address=\"10.1161/CIRCRESAHA.117.312513\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eReveals novel immunological mechanisms and cell type-specific pathways and establishes the functional relevance of lesional leukocytes in human atherosclerosis.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003e\u0026bull;\u003c/b\u003e Yu L, Zhang Y, Liu C and et al. Heterogeneity of macrophages in atherosclerosis revealed by single-cell RNA sequencing. 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AJMBES. 2025;27 (3-4):\u003cspan dir=\"RTL\"\u003e \u003c/span\u003e151-162. http://doi.org/10.53550/AJMBES.2025.v27i03-04.004\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Atherosclerosis, Various macrophages, Single cell transcription, Phenotypic diversity, myocardial infarction","lastPublishedDoi":"10.21203/rs.3.rs-7393034/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7393034/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003ePurpose of Review: \u003c/strong\u003eIn this research, the idea of ​​single cell transcription was used to describe atherosclerotic plaque macrophages.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRecent Findings: \u003c/strong\u003eThe results of the single-cell studies provided detailed data on the transcriptional and phenotypic diversity of macrophages in plaques. Macrophages are regulated by a variety of factors, including cell interactions, oxygen levels, nutrients, metabolites or other soluble signals, and (combination or alteration of) the extracellular matrix. These signals can influence macrophage selection and survival, as well as differentiation and polarization. In addition, macrophages in these plaques were replaced by activated phenotypes containing subsets associated with plaque vulnerability.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSummary: \u003c/strong\u003eAtherosclerosis is due to the multifaceted contribution of the immune system to the traffic and vascular homes. However, the specific properties of altered immune cells in atherosclerotic lesions that lead to clinical events such as ischemic stroke and myocardial infarction are not well-defined. Therefore, our approach provides a powerful tool to help study the underlying mechanisms of human diseases and discover novel pharmacological approaches for therapeutic interventions.\u003c/p\u003e","manuscriptTitle":"A Review of Macrophage Diversity and Immune Checkpoint Regulation in Atherosclerosis: Insights from Single-Cell Transcriptomes","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-12 09:44:30","doi":"10.21203/rs.3.rs-7393034/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"30a41799-2e35-4026-8f81-4f4127c4c0b2","owner":[],"postedDate":"February 12th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-02-12T09:44:30+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-12 09:44:30","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7393034","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7393034","identity":"rs-7393034","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}