Assessment of Inhalation Exposures to Indoor and Occupational Aerosols - Murine Models of Repeated Fungal Inhalation Exposure

室内和职业气溶胶吸入暴露的评估 - 反复真菌吸入暴露的小鼠模型

• 批准号: 10710285
• 负责人: Donald Beezhold
• 金额: $ 36.34万
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 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/10710285
• 关键词: Acoustics; Address; Aerosols; Agreement; Aspergillus; Aspergillus fumigatus; B-Lymphocytes; Basidiomycota; Biological Markers; CD4 Positive T Lymphocytes; Cardiac; Cardiopulmonary; Cells; Cellular Infiltration; Collaborations; Computers; Cryptococcus; Cryptococcus neoformans; Data; Data Set; Development; Disease; EFRAC; Environment; Exposure to; Expression Profiling; Flow Cytometry; Future; Genetic Transcription; Genomics; Goals; Growth; Health; Health Hazards; Heart Rate; IL4 gene; Immune; Immunologics; Impairment; Individual; Indoor environment; Infiltration; Inhalation; Inhalation Exposure; Interleukin-13; Knockout Mice; Knowledge; Lung; Lung immune response; Lymphoid; Lymphoid Cell; Measurement; Measures; Messenger RNA; Methodology; MicroRNAs; Mission; Modeling; Molds; Mouse Strains; Mus; Mycotoxins; Myelogenous; National Institute of Environmental Health Sciences; Natural Immunity; Neurologic; Occupational; Pathology; Play; Production; Proteomics; Public Health; Pulmonary arterial remodeling; Reagent; Recovery; Reporting; Role; Source; Stachybotrys; Study models; System; T-Lymphocyte; Time; Tissues; Toxicology; Yeasts; air sampling; allergic airway disease; base; built environment; cell type; cytokine; exposed human population; fungus; heart function; improved; indoor air; microbial; mouse model; particle; respiratory; respiratory health; response; systemic toxicity

Adverse health effects have been associated with microbial growth and damp indoor environments; however, knowledge gaps exist as to the mechanisms underlying the responses to fungal exposure. To address these knowledge gaps, NIOSH, in collaboration with the NTP and NIEHS, conducted subchronic inhalation studies to examine the pulmonary immunological and toxicological effects following repeated exposure to Aspergillus fumigatus, Stachybotrys chartarum, and Aspergillus versicolor. A computer-controlled acoustical generator system (AGS), based on a modified acoustically powered particle (Pitt-3) generator, was utilized for these studies and models a natural human exposure one would encounter in a fungal contaminated environment. The overall goal of these studies is to characterize the toxicological and pulmonary responses associated with repeated exposure to fungi commonly found in damp, indoor environments. Previously, a large study assessing the pulmonary and systemic toxicity following a 1-, 2-, 3-, 4-, 8-, and 13-week repeated exposure to Aspergillus versicolor was completed. Data showed increased innate immunity cells after 1 week of repeated exposure followed by the increasing infiltration of additional B-cells, T-cells, and type 2 innate lymphoid cells (ILC2s) over 4 weeks. Repeated exposure to A. versicolor led to the increased production of local and circulating Th2 cytokines, including IL4 and IL13, as well increased ILC2s by 4 weeks. By 13 weeks, cellular infiltration was decreased for all cell types except ILC2s. Analysis of miRNA, mRNA, and proteomic datasets derived from this study is ongoing. Similar to the reported responses following A. fumigatus and S. chartarum exposure, pulmonary arterial tissue remodeling was observed following repeated subchronic exposure to A. versicolor. Recovery of pulmonary arterial remodeling was also assessed in FY22, in which mice were repeatedly exposed to A. versicolor for 4 and 13 weeks and then allowed to recover for varying intervals of 1 week to 4 weeks. Results did not show any recovery of the pulmonary arterial tissues. Additional recovery studies are planned for FY23. In FY22, additional exposure studies utilizing genetically modified mouse strains were initiated to further characterize specific mechanisms influencing the pulmonary arterial remodeling observed in previous IAA studies following exposure to A. fumigatus, S. chartarum, and to A. versicolor. Specifically, expression of critical Th2 cytokines known to be involved in pulmonary arterial remodeling, IL4 and IL13, have individually been eliminated in mouse models that are then being used in S. chartarum and A. versicolor exposure studies. Cardiac and respiratory functional measurements are being collected, as well as additional study endpoints including pathology, immune cell profiling via flow cytometry, cytokine quantification, and proteomic and RNA expression profiling. Preliminary results from these studies showed that fungal exposure impaired cardiac function as measured by heart rate and ejection fraction, both of which were restored in the IL13 knockout mouse. Previous sequencing-based studies conducted at NIOSH have also identified fungal yeasts as sources of personal exposure. Basidiomycota yeasts are prominent in indoor contaminated environments, but their role in adverse health effects has remained relatively uncharacterized. To address this knowledge gap, studies in which mice were repeatedly exposed to Cryptococcus victoriae were completed in FY22. The resultant pulmonary immune response was compared to the response elicited from exposure to Cryptococcus neoformans, a yeast known to exacerbate allergic airway disease. Results showed that while both C. neoformans and V. victoriae cells were detectable in the lungs 21 days post final exposure, repeated C. neoformans exposure initiated myeloid and lymphoid cellular infiltration into the lung that worsened over time, whereas repeated V. victoriae exposure induced a strong CD4+ T cell-driven lymphoid response that started to resolve by 21 days post final exposure. Results from these studies provide a better understanding of the role that fungal yeasts play in respiratory health disease. A 13-week study examining the toxicological responses to repeated A. versicolor exposure concluded in FY22. Future studies will investigate endpoints such as cardiopulmonary and neurological responses to fungal exposure.

有害健康影响与微生物生长和潮湿的室内环境有关;然而，关于真菌暴露反应的机制存在知识空白。为了解决这些知识差距，NIOSH与NTP和NIEHS合作，进行了亚慢性吸入研究，以检查重复暴露于烟曲霉、Stachyboischartarum和杂色曲霉后的肺部免疫学和毒理学效应。一个计算机控制的声学发生器系统（AGS），基于一个修改的声学动力粒子（皮特-3）发生器，用于这些研究和模型的自然人类接触一个会遇到的真菌污染的环境。这些研究的总体目标是表征与反复暴露于潮湿室内环境中常见的真菌相关的毒理学和肺部反应。此前，完成了一项大型研究，评估了1周、2周、3周、4周、8周和13周重复暴露于杂色曲霉菌后的肺和全身毒性。数据显示，重复暴露1周后先天免疫细胞增加，随后在4周内增加额外的B细胞、T细胞和2型先天淋巴细胞（ILC 2）的浸润。反复暴露于A。在4周时，杂色导致局部和循环Th 2细胞因子（包括IL 4和IL 13）的产生增加，以及ILC 2增加。到13周时，除ILC 2外，所有细胞类型的细胞浸润均减少。来自本研究的miRNA、mRNA和蛋白质组数据集的分析正在进行中。与A. fumigatus和S.结果表明，在亚慢性暴露A.变色。肺动脉重塑的恢复也在FY 22中进行了评估，其中小鼠反复暴露于A。变色4和13周，然后允许恢复1周至4周的不同间隔。结果未显示肺动脉组织的任何恢复。计划在2023财年进行额外的恢复研究。在2022财年，启动了使用转基因小鼠品系的额外暴露研究，以进一步表征在先前的IAA研究中观察到的暴露于A后影响肺动脉重塑的特定机制。fumigatus、烟曲霉S. chartarum和A.变色。具体地说，已知参与肺动脉重塑的关键Th 2细胞因子IL 4和IL 13的表达在小鼠模型中已经被单独消除，然后将其用于S. chartarum和A.变色暴露研究。正在收集心脏和呼吸功能测量结果，以及其他研究终点，包括病理学、通过流式细胞术进行的免疫细胞分析、细胞因子定量以及蛋白质组学和RNA表达分析。这些研究的初步结果表明，真菌暴露损害了心脏功能，如通过心率和射血分数测量的，这两者在IL 13敲除小鼠中都得到了恢复。 先前在NIOSH进行的基于测序的研究也将真菌酵母确定为个人暴露的来源。担子菌酵母菌在室内污染环境中很突出，但它们在不良健康影响中的作用仍然相对不确定。为了解决这一知识缺口，2022财年完成了小鼠反复暴露于维多利亚隐球菌的研究。将由此产生的肺部免疫应答与暴露于新型隐球菌（一种已知会加重过敏性气道疾病的酵母）引起的应答进行比较。结果表明，C.在最终暴露后21天，在肺中可检测到新生儿和维多利亚弧菌细胞，重复C.新型弧菌暴露引发髓样和淋巴样细胞浸润到肺中，并随时间推移而恶化，而维多利亚弧菌重复暴露诱导强烈的CD 4 + T细胞驱动的淋巴样应答，并在最终暴露后21天开始消退。这些研究的结果使我们更好地了解真菌酵母在呼吸道健康疾病中所发挥的作用。一项为期13周的研究考察了对重复A.在FY 22结束的多色曝光。未来的研究将调查终点，如心肺和神经系统对真菌暴露的反应。