BAFF as a novel SASP factor and regulator of senescent traits

BAFF 作为一种新型 SASP 因子和衰老特征的调节因子

• 批准号: 10471678
• 负责人: Myriam Gorospe
• 金额: $ 16.66万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10471678
• 关键词: Age; Anti-Inflammatory Agents; Autoimmune Diseases; Automobile Driving; B-Lymphocytes; Binding Sites; Blood Vessels; Cell Line; Cell model; Cells; Chronic; Colon Carcinoma; Disease; Evaluation; Family; Fibroblasts; Genetic Transcription; Growth Factor; Human; IL8 gene; IRF1 gene; Immune; Inflammation; Inflammatory; Inflammatory Response; Interferon Type I; Interferons; Interleukin-1; Interleukin-10; Interleukin-6; Metalloproteases; Modeling; Molecular Analysis; Mus; Myeloid Cells; Pathway interactions; Phenotype; Play; Process; Production; Proteins; Regulation; Role; Sampling; Serum; Signal Pathway; TNF gene; TNFRSF5 gene; Tissues; Transcript; Tumor Necrosis Factor Superfamily Ligands; Work; activating transcription factor; age related; aged; belimumab; cancer cell; chemokine; cytokine; in vivo; leukemia; mRNA Precursor; member; monocyte; novel; paracrine; promoter; recruit; response; senescence; trait; transcription factor

ONGOING WORK AIM 1. BAFF regulation in senescence. We confirmed that the cytokine BAFF (Steri M et al., 2017) is elevated in multiple models of senescence. Due to its high expression in myeloid cell lines, we focused on BAFF production in THP-1 (human monocytic) cells for molecular analysis. We found that BAFF is transcriptionally induced in senescent THP-1 cells with enhanced levels of BAFF pre-mRNA. We used ECRbase (https://ecrbase.dcode.org) to identify transcription factors potentially driving BAFF transcription and found that BAFF promoter harbors several binding sites for members of the Interferon Regulated Factor (IRF) transcription factor family (Sjstrand M et al., 2016). Moreover, IRF1 silencing strongly reduced BAFF pre-mRNA and BAFF protein levels in IR-induced senescence, suggesting that BAFF is transcriptionally regulated by IRF1 in senescent cells as a part of the Interferon Type I response. AIM 2. To study BAFF signaling pathways in senescence including SASP. In order to analyze the impact of BAFF on senescence, we silenced BAFF in THP-1 cells and then implemented IR-induced senescence. We observed that BAFF silencing decreased SA-betaGal activity. Interestingly, silencing BAFF decreased SASP factors including the pro-inflammatory cytokines IL-8, IL-6, IL-1/, and TNF and the anti-inflammatory cytokine IL-10. These findings prompted us to analyze NF-B pathway, a major regulator of the inflammatory responses. We observed that BAFF silencing decreased the levels of p50 and p52, representing the canonical and non-canonical NF-B pathways, respectively. Accordingly, the level of several transcripts encoding cytokines and chemokines also decreased. These findings suggest that BAFF may act as an upstream regulator of the SASP, at least in part by acting through the NF-B pathway. FUTURE PLANS In-depth analysis of BAFF functions in senescence. Neutralization of BAFF using Belimumab (Blair et al, 2018) in senescence and evaluation of SASP levels. Quantification of circulating BAFF in human serum as a function of age in GESTALT samples.

正在进行的工作 AIM 1. BAFF在衰老中的调节。 我们证实了细胞因子BAFF（Steri M等人，2017）在多种衰老模型中升高。 由于其在骨髓细胞系中的高表达，我们专注于在THP-1（人单核细胞）细胞中的BAFF产生用于分子分析。 我们发现BAFF在衰老的THP-1细胞中被转录诱导，BAFF前体mRNA水平增加。 我们使用ECRbase（https：//www.example.com）来鉴定可能驱动BAFF转录的转录因子，并发现BAFF启动子具有干扰素调节因子（IRF）转录因子家族成员的几个结合位点（Sjstrand M等人，2016年）。 此外，IRF1沉默强烈地降低了IR诱导的衰老中BAFF前mRNA和BAFF蛋白水平，表明BAFF在衰老细胞中作为I型干扰素应答的一部分由IRF1转录调节。 AIM 2.研究BAFF信号通路在衰老过程中的作用。 为了分析BAFF对衰老的影响，我们在THP-1细胞中沉默BAFF，然后实施IR诱导的衰老。 我们观察到BAFF沉默降低SA-β Gal活性。 有趣的是，沉默BAFF减少了SASP因子，包括促炎细胞因子IL-8、IL-6、IL-1/和TNF以及抗炎细胞因子IL-10。 这些发现促使我们分析NF-B通路，这是炎症反应的主要调节因子。 我们观察到BAFF沉默降低了分别代表经典和非经典NF-B途径的p50和p52的水平。 因此，编码细胞因子和趋化因子的几种转录物的水平也降低。 这些发现表明BAFF可能作为SASP的上游调节因子，至少部分通过NF-B途径起作用。 未来计划 深入分析BAFF在衰老中的功能。 在衰老和SASP水平评价中使用贝利木单抗中和BAFF（Blair et al，2018）。 在GESTALT样品中，作为年龄函数的人血清中循环BAFF的定量。