Distinct Cohorts of Aspergillus fumigatus Transcription Factors Are Required for Epithelial Damage Occurring via Contact- or Soluble Effector-Mediated Mechanisms.
Distinct Cohorts of Aspergillus fumigatus Transcription Factors Are Required for Epithelial Damage Occurring via Contact- or Soluble Effector-Mediated Mechanisms.
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DOI：
10.3389/fcimb.2022.907519
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发表时间：
2022
影响因子：
5.7
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中科院分区：
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肺上皮损伤是由腐殖真菌烟曲霉引起的疾病的统一特征。然而，这种损害的机制基础和监管控制的特点很差。以前的研究已经确定了A.烟曲霉介导的致病性发生在真菌与上皮细胞相互作用的早期（≤ 16小时）或晚期（>16小时），并且分别涉及与宿主细胞的直接接触或由成熟真菌菌丝产生的可溶性因子的作用。上皮损伤的早期和晚期都受到pH响应性转录因子PacC的遗传调控。本研究试图确定是否有其他转录调节因子在调节上皮损伤中发挥作用。特别是，上皮损伤的早期和晚期阶段是否由相同或不同的调节剂控制。此外，是否过程，如孢子吸收和菌丝粘附，以前已被证明是促进上皮损伤，是由相同的上皮调节剂的队列。利用新近构建的A.烟曲霉转录因子无效突变体，进行评估上皮细胞脱离和上皮细胞裂解的两个高通量筛选。共有17个转录因子突变体被发现表现出可重复的赤字上皮损伤的原因。其中，10个突变体在通过上皮脱离引起早期损伤方面有缺陷，8个突变体在通过上皮溶解引起晚期损伤方面有缺陷。值得注意的是，只有一个转录因子，PacC，需要引起上皮损伤的两个阶段。这17个突变体表现出不同的，往往是独特的表型配置文件方面的健身，上皮细胞粘附，细胞壁缺陷，并由上皮细胞的孢子摄取率。引人注目的是，10个突变体中的9个缺陷导致早期损伤也表现出降低的菌丝延伸率，和培养上清液中的8个突变体中的7个缺陷的晚期损伤的细胞毒性显着降低。我们的研究提供了第一个高层次的概述A。烟曲霉调节基因管理肺上皮损伤，表明高度协调的宿主损伤活动，在空间和时间上管理上皮损伤的基因编排。
Damage to the lung epithelium is a unifying feature of disease caused by the saprophytic fungus Aspergillus fumigatus. However, the mechanistic basis and the regulatory control of such damage is poorly characterized. Previous studies have identified A. fumigatus mediated pathogenesis as occurring at early (≤ 16 hours) or late (>16 hours) phases of the fungal interaction with epithelial cells, and respectively involve direct contact with the host cell or the action of soluble factors produced by mature fungal hyphae. Both early and late phases of epithelial damage have been shown to be subject to genetic regulation by the pH-responsive transcription factor PacC. This study sought to determine whether other transcriptional regulators play a role in modulating epithelial damage. In particular, whether the early and late phases of epithelial damage are governed by same or distinct regulators. Furthermore, whether processes such as spore uptake and hyphal adhesion, that have previously been documented to promote epithelial damage, are governed by the same cohorts of epithelial regulators. Using 479 strains from the recently constructed library of A. fumigatus transcription factor null mutants, two high-throughput screens assessing epithelial cell detachment and epithelial cell lysis were conducted. A total of 17 transcription factor mutants were found to exhibit reproducible deficits in epithelial damage causation. Of these, 10 mutants were defective in causing early phase damage via epithelial detachment and 8 mutants were defective in causing late phase damage via epithelial lysis. Remarkably only one transcription factor, PacC, was required for causation of both phases of epithelial damage. The 17 mutants exhibited varied and often unique phenotypic profiles with respect to fitness, epithelial adhesion, cell wall defects, and rates of spore uptake by epithelial cells. Strikingly, 9 out of 10 mutants deficient in causing early phase damage also exhibited reduced rates of hyphal extension, and culture supernatants of 7 out of 8 mutants deficient in late phase damage were significantly less cytotoxic. Our study delivers the first high-level overview of A. fumigatus regulatory genes governing lung epithelial damage, suggesting highly coordinated genetic orchestration of host-damaging activities that govern epithelial damage in both space and time.