{"title":"杂合子错义CSF1R变体阻碍体外CD34+衍生树突状细胞的生成,但不影响体内树突状细胞的发育","authors":"","doi":"10.1016/j.molimm.2024.08.002","DOIUrl":null,"url":null,"abstract":"<div><p>Colony stimulating factor 1 receptor (CSF1R) is an essential receptor for both colony stimulating factor 1 (CSF1) and interleukin (IL) 34 signaling expressed on monocyte precursors and myeloid cells, including monocytes, dendritic cells (DC), and microglia. In humans, dominant heterozygous pathogenic variants in <em>CSF1R</em> cause a neurological condition known as CSF1R-related disorder (CSF1R-RD), typically with late onset, previously referred to as adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP). CSF1R-RD is characterized by microglia reduction and altered monocyte function; however, the impact of pathogenic <em>CSF1R</em> variants on the human DC lineage remains largely unknown.</p><p>We previously reported that cord blood CD34+ stem cell-derived DCs generated <em>in vitro</em> originate specifically from CSF1R expressing precursors. In this study, we examined the DC lineage of four unrelated patients with late-onset CSF1R-RD who carried heterozygous missense <em>CSF1R</em> variants (c.2330G>A, c.2375C>A, c.2329C>T, and c.2381T>C) affecting different amino acids in the protein tyrosine kinase domain of CSF1R. CD34+ stem cells and CD14+ monocytes were isolated from peripheral blood and subjected to an <em>in vitro</em> culture protocol to differentiate towards conventional DCs and monocyte-derived DCs, respectively. Flow cytometric analysis revealed that monocytes from patients with late-onset CSF1R-RD were still able to differentiate into monocyte-derived DCs <em>in vitro</em>, whereas the ability of CD34+ stem cells to differentiate into conventional DCs was impaired. Strikingly, the peripheral blood of patients contained all naturally occurring DC subsets. We conclude that the <em>in vitro</em> abrogation of DC-development in patients with heterozygous pathogenic missense <em>CSF1R</em> variants does not translate to an impairment in DC development <em>in vivo</em> and speculate that CSF1R signalling <em>in vivo</em> is compensated, which needs further study.</p></div>","PeriodicalId":18938,"journal":{"name":"Molecular immunology","volume":null,"pages":null},"PeriodicalIF":3.2000,"publicationDate":"2024-08-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0161589024001585/pdfft?md5=0ee273246aee58fdb03ae182cfc87876&pid=1-s2.0-S0161589024001585-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Heterozygous missense CSF1R variants hamper in vitro CD34+-derived dendritic cell generation but not in vivo dendritic cell development\",\"authors\":\"\",\"doi\":\"10.1016/j.molimm.2024.08.002\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Colony stimulating factor 1 receptor (CSF1R) is an essential receptor for both colony stimulating factor 1 (CSF1) and interleukin (IL) 34 signaling expressed on monocyte precursors and myeloid cells, including monocytes, dendritic cells (DC), and microglia. In humans, dominant heterozygous pathogenic variants in <em>CSF1R</em> cause a neurological condition known as CSF1R-related disorder (CSF1R-RD), typically with late onset, previously referred to as adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP). CSF1R-RD is characterized by microglia reduction and altered monocyte function; however, the impact of pathogenic <em>CSF1R</em> variants on the human DC lineage remains largely unknown.</p><p>We previously reported that cord blood CD34+ stem cell-derived DCs generated <em>in vitro</em> originate specifically from CSF1R expressing precursors. In this study, we examined the DC lineage of four unrelated patients with late-onset CSF1R-RD who carried heterozygous missense <em>CSF1R</em> variants (c.2330G>A, c.2375C>A, c.2329C>T, and c.2381T>C) affecting different amino acids in the protein tyrosine kinase domain of CSF1R. CD34+ stem cells and CD14+ monocytes were isolated from peripheral blood and subjected to an <em>in vitro</em> culture protocol to differentiate towards conventional DCs and monocyte-derived DCs, respectively. Flow cytometric analysis revealed that monocytes from patients with late-onset CSF1R-RD were still able to differentiate into monocyte-derived DCs <em>in vitro</em>, whereas the ability of CD34+ stem cells to differentiate into conventional DCs was impaired. Strikingly, the peripheral blood of patients contained all naturally occurring DC subsets. We conclude that the <em>in vitro</em> abrogation of DC-development in patients with heterozygous pathogenic missense <em>CSF1R</em> variants does not translate to an impairment in DC development <em>in vivo</em> and speculate that CSF1R signalling <em>in vivo</em> is compensated, which needs further study.</p></div>\",\"PeriodicalId\":18938,\"journal\":{\"name\":\"Molecular immunology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.2000,\"publicationDate\":\"2024-08-24\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S0161589024001585/pdfft?md5=0ee273246aee58fdb03ae182cfc87876&pid=1-s2.0-S0161589024001585-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Molecular immunology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0161589024001585\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular immunology","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0161589024001585","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
集落刺激因子 1 受体(CSF1R)是集落刺激因子 1(CSF1)和白细胞介素(IL)34 信号转导的重要受体,表达于单核细胞前体和髓系细胞,包括单核细胞、树突状细胞(DC)和小胶质细胞。在人类中,CSF1R 的显性杂合子致病变体会导致一种被称为 CSF1R 相关障碍(CSF1R-RD)的神经系统疾病,通常发病较晚,以前被称为伴有轴突球和色素性胶质细胞的成人发病型白质脑病(ALSP)。CSF1R-RD的特征是小胶质细胞减少和单核细胞功能改变;然而,致病性CSF1R变体对人类DC系的影响在很大程度上仍是未知的。我们以前曾报道,体外生成的脐带血CD34+干细胞衍生DC特异性地来源于CSF1R表达的前体。在这项研究中,我们检测了四名无血缘关系的晚发性CSF1R-RD患者的直流血统,这些患者携带杂合子错义CSF1R变体(c.2330G>A、c.2375C>A、c.2329C>T和c.2381T>C),这些变体影响了CSF1R蛋白酪氨酸激酶结构域中的不同氨基酸。从外周血中分离出 CD34+ 干细胞和 CD14+ 单核细胞,并对其进行体外培养,使其分别向传统 DC 和单核细胞衍生 DC 分化。流式细胞仪分析表明,晚发型CSF1R-RD患者的单核细胞仍能在体外分化为单核细胞源性直流细胞,而CD34+干细胞分化为常规直流细胞的能力受损。令人震惊的是,患者外周血中含有所有自然产生的 DC 亚群。我们的结论是,杂合子致病性CSF1R错义变体患者体内DC发育的体外衰减并不转化为体内DC发育的障碍,并推测CSF1R信号在体内可得到补偿,这需要进一步研究。
Heterozygous missense CSF1R variants hamper in vitro CD34+-derived dendritic cell generation but not in vivo dendritic cell development
Colony stimulating factor 1 receptor (CSF1R) is an essential receptor for both colony stimulating factor 1 (CSF1) and interleukin (IL) 34 signaling expressed on monocyte precursors and myeloid cells, including monocytes, dendritic cells (DC), and microglia. In humans, dominant heterozygous pathogenic variants in CSF1R cause a neurological condition known as CSF1R-related disorder (CSF1R-RD), typically with late onset, previously referred to as adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP). CSF1R-RD is characterized by microglia reduction and altered monocyte function; however, the impact of pathogenic CSF1R variants on the human DC lineage remains largely unknown.
We previously reported that cord blood CD34+ stem cell-derived DCs generated in vitro originate specifically from CSF1R expressing precursors. In this study, we examined the DC lineage of four unrelated patients with late-onset CSF1R-RD who carried heterozygous missense CSF1R variants (c.2330G>A, c.2375C>A, c.2329C>T, and c.2381T>C) affecting different amino acids in the protein tyrosine kinase domain of CSF1R. CD34+ stem cells and CD14+ monocytes were isolated from peripheral blood and subjected to an in vitro culture protocol to differentiate towards conventional DCs and monocyte-derived DCs, respectively. Flow cytometric analysis revealed that monocytes from patients with late-onset CSF1R-RD were still able to differentiate into monocyte-derived DCs in vitro, whereas the ability of CD34+ stem cells to differentiate into conventional DCs was impaired. Strikingly, the peripheral blood of patients contained all naturally occurring DC subsets. We conclude that the in vitro abrogation of DC-development in patients with heterozygous pathogenic missense CSF1R variants does not translate to an impairment in DC development in vivo and speculate that CSF1R signalling in vivo is compensated, which needs further study.
期刊介绍:
Molecular Immunology publishes original articles, reviews and commentaries on all areas of immunology, with a particular focus on description of cellular, biochemical or genetic mechanisms underlying immunological phenomena. Studies on all model organisms, from invertebrates to humans, are suitable. Examples include, but are not restricted to:
Infection, autoimmunity, transplantation, immunodeficiencies, inflammation and tumor immunology
Mechanisms of induction, regulation and termination of innate and adaptive immunity
Intercellular communication, cooperation and regulation
Intracellular mechanisms of immunity (endocytosis, protein trafficking, pathogen recognition, antigen presentation, etc)
Mechanisms of action of the cells and molecules of the immune system
Structural analysis
Development of the immune system
Comparative immunology and evolution of the immune system
"Omics" studies and bioinformatics
Vaccines, biotechnology and therapeutic manipulation of the immune system (therapeutic antibodies, cytokines, cellular therapies, etc)
Technical developments.