基于高通量测序解析眼部感染致病真菌对不同波长可见光刺激耐受性的研究：眼部真菌感染发病与防御机制的探讨

Fungal infections of the eye are considered one of the most important cause of blindness. The mechanism of fungal invasion of ocular tissue, cornea destruction and dysfunction of immune system has not been elucidated. Previous researches demonstrated that Toll like receptors and Dectin-1, a receptor for the major fungal cell wall carbohydrate β-1,3 glucan, may involved in the inflammatory response and restoration to fungal infections. The effect of light regulation on fungi’s growth, development and secondary metabolism of fungi has been widely used. But it is still unclear whether light has the same effect on intraocular infectious fungi. In the earlier period of our study, we plan to divide visible light into different wave-length, and observe the injury caused by different narrow-band visible light and the following cell protection. In order to find out different wave-lengths and intensities visible light’s effect on the eye’s infection fungi’s genome information and transcriptome traits, we plan to use a illumination system of microorganism using lights of different wave-lengths and intensities, the hight-throughput sequencing technology, and use the common ocular infection fungi which obtained from clinical cases, such as Fusarium, Penicillium, Aspergillus and Saccharomyces.

真菌感染性眼病是目前最为严重的致盲性眼部疾病之一。目前对眼部真菌侵袭眼部组织、破坏角膜和眼部组织天然免疫功能的致病机制尚不清楚。前期研究证实，免疫系统包括Toll样受体和β-葡聚糖特异性识别受体Dectin-1可能参与真菌感染后组织反应与损伤修复。光对真菌生长发育、生理周期、形态变化及次生代谢产物的调控已被广泛利用，但光对眼部感染致病真菌是否有同样的调控机制尚无研究。我们前期的工作主要分别聚焦于拆分可见光，观察不同窄谱可见光光损伤及眼部细胞保护。本课题拟利用不同波长和强度的微生物光作用系统、高通量测序技术，以临床样本获得的常见眼部感染致病镰刀菌属、青霉曲霉菌属、酵母菌属为研究对象，从全基因组、转录组、细胞生物学水平，探讨不同波长和强度可见光作用下眼部感染致病真菌的基因组遗传信息和转录组特征，揭示眼部感染致病真菌可能的致病相关机理。

本课题以临床获得的常见眼部感染致病镰刀菌、曲霉菌、酵母菌为主要研究对象，以实验室标准菌株作为对照，利用不同波长和强度的微生物光作用系统、微生物培养系统、高通量测序技术，从真菌生物学特性、基因组、转录组等方面，探讨不同外界营养环境、不同强度可见光作用下眼部感染致病真菌的致病性相关特征变化。研究发现，眼部致病真菌对氮源对利用有选择倾向性，角膜含量较多的甘氨酸、脯氨酸的利用可能与致病真菌的致病性密切相关；蓝光对角膜致病茄病镰刀菌的调控存在两面性，一方面抑制菌丝的生长，另一方面促进孢子的形成。本研究为进一步探索营养环境及可见光对眼部致病真菌的分子调控机制打下坚实基础，这对我们未来探究治疗真菌性角膜炎的新途径有着重要意义。项目资助发表SCI论文2篇，核心期刊论文1篇。培养博士后2名，博士1名，硕士1名。项目投入经费60万，支出58.15万，各项支出基本与预算相符。