Molecular Characterization Of Fungal Biofilm Formation

描述：该R21的目的是表征从C.白色念珠菌通过发育成熟和连续的细胞网络粘附到非生物表面。我们的假设是，在C.白念珠菌生物膜将导致在该过程中表达的关键阶段、途径和功能的鉴定。在细菌和真菌中的生物膜形成过程主要通过在微观和宏观水平上描述导致在惰性表面上建立生物膜的事件来评估。在真菌生物膜的情况下，很少有人知道与生物膜的发展相关的潜在的分子过程。除了有限的生物化学表征的细胞外基质，药物敏感性的变化，在生物膜中的细胞和主要与菌丝形态发生的调节相关的各种突变的影响，很少有人知道的分子变化，这可能是表现在粘附细胞相比，粘附细胞。关于分子变化知之甚少，这些分子变化可能发生在贴壁细胞中，因为这些细胞继续建立高度结构化的细胞网络。在这个高通量基因组分析的时代，我们有独特的能力来解决这个新出现的问题，通过使用的方法，允许在生物膜的发展过程中的基因表达的变化进行全面的分析。在这项研究中，我们将使用我们与斯坦福大学基因组技术中心（STGC）的罗纳德戴维斯博士及其同事开发的DNA微阵列;这些阵列包括C.白色念珠菌RNA样品将在高度受控的时间过程中从贴壁和非贴壁细胞中分离，该时间过程将跨越在本申请中描述的条件下形成生物膜所需的24小时时间。将使用各种算法仔细分析将从这样的研究中出现的大数据集，这些算法将允许定义特定基因和基因簇/途径中的变化，这些变化可能在生物膜发育过程中表达。通过比较贴壁和非贴壁细胞中的基因表达，我们将了解是否以及在何种程度上，附着到惰性表面可以影响基因表达。

DESCRIPTION: The purpose of this R21 is to characterize the molecular events in fungal biofilm formation from the initial attachment of C. albicans to an abiotic surface through the development of a mature and continuous cellular network that is firmly adherent to that surface. It is our hypothesis that transcription profiling during the development of a C. albicans biofilm will result in the identification of key stages, pathways and functions that are expressed during this process. The process of biofilm formation in both bacteria and fungi has primarily been assessed through the description of events, at the microscopic and macroscopic level, which result in the establishment of biofilms on inert surfaces. In the case of fungal biofilms, little is known about the underlying molecular process associated with biofilm development. With the exception of limited biochemical characterization of the extracellular matrix, the changes in drug susceptibility in cells in biofilms and the effects of various mutations associated primarily with the regulation of hyphal morphogenesis, little is known about the molecular changes which may be manifest in adherent cells as compared with planktonic cells. Less is known about the molecular changes, which may occur in adherent cells as these proceed to establish highly-structured cellular networks. In this era of high throughput genome profiling, we have the unique ability to address this emerging problem by using methods that allow comprehensive analysis of the changes in gene expression during the course of biofilm development. For this study, we will use DNA microarrays, which we have developed with Dr. Ronald Davis and colleagues at the Stanford Genome Technology Center (STGC); these arrays are inclusive of the entire genome of C. albicans. RNA samples will be isolated from adherent and planktonic cells in a highly-controlled time course which will span the 24h period required for biofilm formation under the conditions described in the application. The large data set that will emerge from such a study will be carefully analyzed using a variety of algorithms that will allow definition of changes in specific genes and in gene clusters/pathways, which may be expressed during the course of biofilm development. By comparing gene expression in adherent and non-adherent cells, we will learn whether and to what degree, attachment to an inert surface can affect gene expression.