Defining the mechanism of skin barrier restoration by bacterial lysates

定义细菌裂解物修复皮肤屏障的机制

• 批准号: 2725954
• 金额: --
• 依托单位: Queen Mary University of London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2725954
• 关键词: Defining; mechanism; skin; barrier; restoration

Background: AKT1 controls multiple aspects of skin barrier formation: Ryan O'Shaughnessy has shown that AKT1 knockdown leads defects in both the physical barrier and immune barrier. It prevents filaggrin processing, and inhibits nuclear degradation, a key part of the formation of a tough, protective cornified envelope. AKT1 knockdown keratinocytes have reduced expression of a protease Cathepsin H (CTSH). CTSH is required for filaggrin processing, and control of Th2 cytokines, with Ctsh -/- mice showing increased expression of the Th2 cytokine TSLP.Effects of bacterial lysates on epidermal barrier: Catherine O'Neill is an expert on the beneficial effects of lactobacillus and bifidobacterial lysates. They enhance tight junctions and promote filaggrin expression, both key components of healthy skin barrier. SB has developed a topical formulation containing one of these bacterial lysates for use in a human study, where they showed reduced transepidermal water loss (an in vivo marker of the barrier) in human volunteers (SB personal communication). The company wishes to understand mechanistically how its lysate is inducing its effects on the barrier. To this end SB developed propriatory techniques to produce lactobacillus lysate fractions and tested them in their bespoke assays.We hypothesise that these lysates have multiple additional beneficial skin barrier restorative functions related to the processes described aboveOverall approach: We will test microbial lysates and fractions thereof developed by SB in barrier deficient and competent keratinocytes and organotypic cultures to understand how they restore or enhance skin barrier. The student will perform proteomics to identify biomarkers of efficacy of the microbial lysates. This work will be critical to SB in their aim of the downstream identification of actives within the microbial preparations, and to skin barrier biology research by improving understanding not only of skin barrier function, but also bacteria-host interaction, currently a "hot-topic" in skin biology research.Objectives:1. Testing the ability of bacterial lysates to rescue physical and immune skin barrier function2. Determining the proteomic profile of keratinocytes treated with bacterial lysates to identify pathways and biomarkers associated with barrier enhancement3. Fractionation of lysates and assaying biomarkers to identify key actives within the bacterial lysates.Interdisciplinary aspect and added value: This project combines cell biology, proteomics/bioinformatics , and SB's unique expertise in microbiology and the use of separation techniques to produce lysates with biological activity and to gain new insights into barrier function and bacteria-host interactions in skin.

背景资料：AKT 1控制皮肤屏障形成的多个方面：Ryan O 'Shaughnessy已经表明，AKT 1敲低导致物理屏障和免疫屏障的缺陷。它阻止聚丝蛋白加工，抑制核降解，这是形成坚韧的保护性皮质包膜的关键部分。AKT 1敲低的角质形成细胞具有蛋白酶组织蛋白酶H（CTSH）的降低的表达。CTSH是加工聚丝蛋白和控制Th 2细胞因子所必需的，Ctsh -/-小鼠显示出Th 2细胞因子TSLP的表达增加。它们增强紧密连接并促进丝聚蛋白表达，这两者都是健康皮肤屏障的关键组成部分。SB开发了一种含有这些细菌裂解物之一的局部制剂，用于人体研究，其中它们显示出人类志愿者经表皮水分损失（屏障的体内标志物）减少（SB个人交流）。该公司希望从机制上了解其裂解物如何诱导其对屏障的影响。为此，SB开发了专有的技术来生产溶胞产物级分，并在其定制的测定中对其进行了测试。我们假设这些溶胞产物具有与上述过程相关的多种额外的有益的皮肤屏障恢复功能。我们将在屏障缺陷和胜任角质形成细胞和器官型培养物中测试SB开发的微生物裂解物及其组分，以了解它们如何恢复或增强皮肤屏障学生将进行蛋白质组学研究，以识别微生物裂解物功效的生物标志物。这项工作将是至关重要的SB在他们的目标的下游识别的活性微生物制剂，皮肤屏障生物学研究，通过提高理解不仅是皮肤屏障功能，但也细菌-宿主相互作用，目前在皮肤生物学研究的“热点”。测试细菌裂解物拯救物理和免疫皮肤屏障功能的能力2。确定用细菌裂解物处理的角质形成细胞的蛋白质组谱，以鉴定与屏障增强相关的途径和生物标志物3。跨学科和附加价值：该项目结合了细胞生物学、蛋白质组学/生物信息学和SB在微生物学方面的独特专业知识，并使用分离技术生产具有生物活性的裂解物，并获得对皮肤屏障功能和细菌-宿主相互作用的新见解。