Cimická Jana, Riegert Jan, Kavková Miloslava, Černá Kateřina
Laboratory of Pain research, Institute of Physiology, Czech Academy of Sciences, 142 20 Prague, Czech Republic.
Department of Molecular Genetics, Bioptická Laboratoř s.r.o., 326 00 Pilsen, Czech Republic.
Med Mycol. 2022 Jan 10;60(1). doi: 10.1093/mmy/myab076.
Analysis of mycobiome from formalin-fixed, paraffin-embedded (FFPE) biopsies should preferentially detect only fungi which are actually present in the intestine wall, in contrast to stool samples, which are limited by the diet composition. Next generation sequencing provides the advantage of analyzing many species from a single sample. Consequently, canonical correspondence analysis divided fungal genera present in FFPE intestinal tissues into three well-defined experimental groups (negative controls - NC, Crohn's disease - CD, ulcerative colitis - UC). Simultaneously, the analysis showed that particular fungal genera are associated with these experimental groups and several fungal genera occurred in all experimental groups equally. Our results also showed a noticeable increase of Ascomycota proportion from NC, through CD to UC. Fungal genera Malassezia, Cladosporium and Toninia occurred in all experimental groups assuming that they are common components of the intestinal mycobiome. Other fungal genera found only in the NC experimental group were non-pathogenic and might bring some benefits. In contrast, CD and UC samples were characterized by an accumulation of genera with inhibitive effects on growth of other fungal genera and the presence of opportunistic pathogens. Furthermore, a decrease in the fungal genus Malassezia in inflammatory tissues was observed; Specifically, the UC experimental group showed a connection between the presence of Candida and seven time's lower amounts of Malassezia (compared to amounts found in NC). The CD experimental group was characterized by the simultaneous presence of Engyodontium album with Lecanicillium, and indicates a possible pathogenic effect of Ramularia in disease development.
Mycobiome analysis of formalin-fixed and paraffin-embedded biopsies may highlight actual fungal genera composition in the intestinal wall. Interestingly, experimental groups of Crohn's disease and ulcerative colitis clearly differed by structure of their mycobiomes.
与受饮食成分限制的粪便样本不同,对福尔马林固定、石蜡包埋(FFPE)活检组织的真菌群落分析应优先仅检测肠壁中实际存在的真菌。新一代测序提供了从单个样本分析多种物种的优势。因此,典范对应分析将FFPE肠道组织中存在的真菌属分为三个明确的实验组(阴性对照-NC、克罗恩病-CD、溃疡性结肠炎-UC)。同时,分析表明特定的真菌属与这些实验组相关,并且有几个真菌属在所有实验组中均有同等程度的出现。我们的结果还显示,从NC组到CD组再到UC组,子囊菌门的比例显著增加。马拉色菌属、枝孢属和托尼菌属在所有实验组中均有出现,表明它们是肠道真菌群落的常见组成部分。仅在NC实验组中发现的其他真菌属无致病性,可能带来一些益处。相比之下,CD组和UC组样本的特征是存在对其他真菌属生长有抑制作用的属以及机会性致病菌。此外,观察到炎症组织中马拉色菌属真菌减少;具体而言,UC实验组显示念珠菌的存在与马拉色菌数量降低七倍之间存在关联(与NC组中的数量相比)。CD实验组的特征是同时存在白内齿菌和 Lecanicillium,并表明柱隔孢属在疾病发展中可能具有致病作用。
对福尔马林固定和石蜡包埋活检组织的真菌群落分析可能突出肠壁中实际的真菌属组成。有趣的是,克罗恩病和溃疡性结肠炎的实验组在其真菌群落结构上明显不同。