The role of beta 2 integrins in macrophage fusion

β2整合素在巨噬细胞融合中的作用

• 批准号: 10545168
• 负责人: Tatiana P Ugarova
• 金额: $ 44.68万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10545168
• 关键词: Actins; Adherens Junction; Adhesives; Adsorption; Affect; Afibrinogenemia; Artificial Implants; Attenuated; Binding Sites; Biocompatible Materials; Biological; Biology; CD18 Antigens; CD47 gene; Cell Survival; Cell membrane; Chronic; Complex; Data; Defect; Deposition; Dissection; E-Cadherin; Endogenous Retroviruses; Enzymes; Epithelium; Event; Extracellular Space; Failure; Fibrin; Fibrinogen; Fluorescence; Foreign Bodies; Foreign-Body Reaction; Gene Targeting; Genes; Giant Cells; Goals; Grant; Granulation Tissue; Imaging Techniques; Immunoelectron Microscopy; Implant; In Vitro; Inflammation; Integrins; Knock-out; Knowledge; Ligands; Macrophage; Macrophage-1 Antigen; Medical Device; Microfluidic Microchips; Microfluidics; Microscopy; Modeling; Molecular; Mus; Myeloid Cells; Nanotechnology; PTPNS1 gene; Phenotype; Plasma Proteins; Polymers; Property; Prosthesis Implantation; Proteins; Proteomics; Recording of previous events; Research; Resolution; Role; Structure; Surface; System; Testing; Thick; Time; Translating; Vascular Graft; Video Microscopy; Wild Type Mouse; adhesion receptor; biomaterial development; experimental study; high resolution imaging; implantation; in vitro Model; in vivo; knock-down; live cell imaging; mechanical properties; mouse model; nectin; novel; polymerization; protein E; receptor; response; syncytin; technological innovation

PROJECT SUMMARY Macrophage fusion resulting in the formation of multinucleated giant cells (MGCs) accompanies a variety of maladies associated with chronic inflammation, including the foreign body response (FBR) elicited by implanted biomaterials. Despite the long history of research on FBR, the molecular and cellular mechanisms of macrophage fusion, an event central to the long-term failure of implanted prosthetic vascular grafts and other medical devices, remain poorly understood. In our preliminary studies using in vivo implantation model, we found that the formation of MGCs and granulation tissue, which develops around the implant and is a precursor of the undesirable fibrotic cap, was almost completely abolished in fibrinogen-deficient mice. Surprisingly, the number of MGCs formed on biomaterials implanted into Mac-1-deficient mice was greater than in wild-type mice and the thickness of granulation tissue was larger. We hypothesize that macrophage fusion on biomaterials critically depends on the deposited fibrin(ogen) matrix and the absence of Mac-1, through the alteration of adhesive properties of macrophages, exacerbates the FBR. Specific Aim 1 is to test this hypothesis. Using a mouse model of biomaterial implantation and gene-targeted mice, we will perform systematic analyses of the early and late stages of FBR and determine the M1/M2 phenotype of MGCs derived from wild-type and Mac-1-deficient macrophages. Using nanotechnology approaches we will characterize the adhesive and mechanical properties of fibrin(ogen) matrices deposited on biomaterials in wild-type and Mac-1-deficient mice. Specific Aim 2 will characterize previously unrecognized actin- based zipper-like structures (ZLS) that form between MGCs on implanted biomaterials. We developed an in vitro model that reproduces the formation of ZLS and demonstrated that the intercellular space within ZLS is filled with junctional proteins E-cadherin and nectin-2. We hypothesize that MGCs form epithelial-like junctions that aid the MGC survival. Taking advantage of technological innovations including a microfluidic chamber that allows the precise dissection of ZLS followed by proteomics analyses, high-resolution microscopy, live cell imaging and mice with myeloid cell-specific KO of E- cadherin and other components of junctions, we will determine the composition of ZLS and their role in the FBR. Specific Aim 3 is to determine the role of authentic fusogenic proteins syncytins in macrophage fusion. Based on our finding that macrophage fusion is initiated by an actin-based protrusion, we will use knockdown experiments, EM and video microscopy to test the hypothesis that a fusion-competent protrusion at the leading edge of a donor macrophage contains syncytins. Overalls, these studies will define the novel biology of macrophage fusion and characterize new mechanisms that have the potential to modulate the FBR.

项目摘要 巨噬细胞融合导致多核巨细胞（MGCs）的形成， 与慢性炎症相关的各种疾病，包括异物反应（FBR） 由植入的生物材料引起。尽管FBR的研究历史悠久，但分子和 巨噬细胞融合的细胞机制，这是植入的巨噬细胞长期失败的核心事件， 人造血管移植物和其他医疗装置仍然知之甚少。在我们的初步调查中 使用体内植入模型的研究，我们发现MGCs和肉芽组织的形成， 其在植入物周围发展并且是不期望的纤维化帽的前体， 在纤维蛋白原缺乏的小鼠中完全消除。令人惊讶的是，MGC形成的数量 植入Mac-1缺陷小鼠的生物材料的厚度大于野生型小鼠， 肉芽组织的体积较大。我们假设巨噬细胞在生物材料上的融合 依赖于沉积的纤维蛋白（原）基质和Mac-1的缺乏，通过改变 巨噬细胞的粘附特性，加剧了FBR。具体目标1是检验这一假设。 使用生物材料植入的小鼠模型和基因靶向小鼠，我们将进行系统的 分析FBR的早期和晚期，并确定M1/M2表型的MGCs衍生物 来自野生型和Mac-1缺陷型巨噬细胞。使用纳米技术的方法，我们将 表征沉积在生物材料上的纤维蛋白（原）基质的粘附和机械性能 在野生型和Mac-1缺陷小鼠中。具体目标2将表征以前未识别的肌动蛋白- 基于拉链状结构（ZLS），其在植入的生物材料上的MGCs之间形成。我们开发 一种体外模型，再现了ZLS的形成，并证明了细胞间隙 在ZLS内充满了连接蛋白E-钙粘蛋白和连接蛋白-2。我们假设MGCs形成于 帮助MGC存活的上皮样连接。利用技术创新 包括一个微流控室，可以精确解剖ZLS，然后进行蛋白质组学研究， 分析，高分辨率显微镜，活细胞成像和具有骨髓细胞特异性KO的E- 我们将确定ZLS的组成及其在连接中的作用， FBR。具体目标3是确定真正的融合蛋白合胞素在 巨噬细胞融合基于我们发现巨噬细胞融合是由一种基于肌动蛋白的 突出，我们将使用击倒实验，EM和视频显微镜来测试假设， 供体巨噬细胞前缘的融合活性突起含有合胞素。工作服， 这些研究将定义巨噬细胞融合的新生物学，并表征新的机制， 具有调节FBR的潜力。

项目成果

The role of integrin alpha D beta2 (CD11d/CD18) in monocyte/macrophage migration.

• DOI: 10.1016/j.yexcr.2008.05.016
• 发表时间: 2008-08
• 期刊: Experimental cell research
• 影响因子: 3.7
• 作者: V. Yakubenko;N. Belevych;D. Mishchuk;A. Schurin;S. Lam;T. Ugarova

A cluster of basic amino acid residues in the gamma370-381 sequence of fibrinogen comprises a binding site for platelet integrin alpha(IIb)beta3 (glycoprotein IIb/IIIa).

• DOI: 10.1021/bi051581d
• 发表时间: 2005-12
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Nataly P. Podolnikova;O. Gorkun;R. Loreth;V. Yee;S. Lord;T. Ugarova

RGD-dependent binding of TP508 to integrin alphavbeta3 mediates cell adhesion and induction of nitric oxide.

• DOI: 10.1160/th09-07-0447
• 发表时间: 2010-07
• 期刊: Thrombosis and haemostasis
• 影响因子: 6.7
• 作者: Derkach DN;Wadekar SA;Perkins KB;Rousseau E;Dreiza CM;Cheung-Flynn J;Ramos HC;Ugarova TP;Sheller MR
• 通讯作者: Sheller MR

Transition of podosomes into zipper-like structures in macrophage-derived multinucleated giant cells.

• DOI: 10.1091/mbc.e19-12-0707
• 发表时间: 2020-08-15
• 期刊: Molecular biology of the cell
• 影响因子: 3.3
• 作者: Balabiyev A;Podolnikova NP;Mursalimov A;Lowry D;Newbern JM;Roberson RW;Ugarova TP
• 通讯作者: Ugarova TP

PLATELET FACTOR 4 (PF4) IMPROVES SURVIVAL IN A MURINE MODEL OF ANTIBIOTIC-SUSCEPTIBLE AND METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS PERITONITIS.

血小板因子 4 (PF4) 可提高抗生素敏感且耐甲氧西林金黄色葡萄球菌腹膜炎小鼠模型的存活率。

• DOI: 10.1101/2023.08.25.554865
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Podolnikova,NatalyP;Lishko,ValeryiK;Roberson,Robert;Koh,Zhqian;Derkach,Dmitry;Richardson,David;Sheller,Michael;Ugarova,TatianaP
• 通讯作者: Ugarova,TatianaP