Understanding regulation of lipid synthesis and signalling in the skin barrier

了解皮肤屏障中脂质合成和信号传导的调节

• 批准号: 1651088
• 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: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1651088
• 关键词: Understanding; regulation; lipid; synthesis; signalling

The applicants plan to take advantage of a unique next-gneration sequencing (RNASeq) data set developed collaboratively with Stiefel/GSK from transient knockdown of transglutaminase 1 (TGM1), ichthyin (NIPAL4) and ALOX12B (Arachidonate 12-lipooxygenase) in differentiated epidermal keratinocytes. These genes are important in maintaining a stable skin barrier. Mutations in these genes cause a severe scaly skin disease called ichthyosis. Stable knockdown 3D models will be developed using a human keratinocyte cell line. Functional studies will be performed to validate the 3D model. The RNASeq data set will be mined looking for novel common and differentiating pathways between the 3 datasets. These will then be validated at the protein level in the 3D models. Lipid profiling and lipidomics will be performed using electrospray ionization tandem mass spectrometry (ESI-MS/MS) and modern bioinformatic software at GSK. Techniques acquired will include cell culture, 3D cultures of skin models, shRNA knock-down using lentivirus, Q-PCR, Western blotting, immunostaining, confocal microscopy, RNAseq and proteomic analysis. The student will also have an opportunity to spend time with the industrial partner.

申请人计划利用与 Stiefel/GSK 合作开发的独特的下一代测序 (RNASeq) 数据集，该数据集通过瞬时敲低分化的表皮角质形成细胞中的转谷氨酰胺酶 1 (TGM1)、鱼腥素 (NIPAL4) 和 ALOX12B（花生四烯酸 12-脂加氧酶）来实现。这些基因对于维持稳定的皮肤屏障很重要。这些基因的突变会导致严重的鳞状皮肤病，称为鱼鳞病。将使用人类角质形成细胞系开发稳定的敲低 3D 模型。将进行功能研究以验证 3D 模型。将挖掘 RNASeq 数据集，寻找 3 个数据集之间新颖的共同和差异路径。然后，这些将在 3D 模型的蛋白质水平上得到验证。脂质分析和脂质组学将使用 GSK 的电喷雾电离串联质谱 (ESI-MS/MS) 和现代生物信息学软件进行。获得的技术包括细胞培养、皮肤模型的 3D 培养、使用慢病毒的 shRNA 敲低、Q-PCR、蛋白质印迹、免疫染色、共聚焦显微镜、RNAseq 和蛋白质组分析。学生还将有机会与工业合作伙伴共度时光。

项目成果

Understanding the role of ABCA12 in the pathogenesis of Harlequin Ichthyosis

了解 ABCA12 在丑角鱼鳞病发病机制中的作用

• 发表时间: 2016
• 期刊: JOURNAL OF INVESTIGATIVE DERMATOLOGY
• 影响因子: 6.5
• 作者: Enjalbert F.

Developmental cell programs are co-opted in inflammatory skin disease.

• DOI: 10.1126/science.aba6500
• 发表时间: 2021-01-22
• 期刊: Science (New York, N.Y.)
• 作者: Reynolds G;Vegh P;Fletcher J;Poyner EFM;Stephenson E;Goh I;Botting RA;Huang N;Olabi B;Dubois A;Dixon D;Green K;Maunder D;Engelbert J;Efremova M;Polański K;Jardine L;Jones C;Ness T;Horsfall D;McGrath J;Carey C;Popescu DM;Webb S;Wang XN;Sayer B;Park JE;Negri VA;Belokhvostova D;Lynch MD;McDonald D;Filby A;Hagai T;Meyer KB;Husain A;Coxhead J;Vento-Tormo R;Behjati S;Lisgo S;Villani AC;Bacardit J;Jones PH;O'Toole EA;Ogg GS;Rajan N;Reynolds NJ;Teichmann SA;Watt FM;Haniffa M
• 通讯作者: Haniffa M