Investigating cellular pathway changes within skin in response to different combinational components of solar light and sunscreen interventions

皮肤老化是一个复杂的过程，由内在和外在因素共同决定，导致结构和功能的渐进性丧失。线粒体是细胞氧化应激的主要来源，广泛参与皮肤衰老，因为它们直接影响皮肤的生物能量，并影响其他细胞过程和核DNA损伤，这也将在皮肤衰老中发挥关键作用。外源性皮肤老化在很大程度上是由环境因素和外部应激源推动的，如紫外线(以及最近太阳光的可见光和红外线成分)，污染和生活方式因素已被证明刺激活性氧的产生和产生氧化应激。这些外源性来源的损害会损害皮肤的结构和功能，导致外源性皮肤衰老的表型特征。研究计划博士计划将应用之前与Croda博士(Catherine Bonn学生)建立的一些技术，利用我们发表在FASEB论文(2020)上的阳光的紫外线、可见光和红外成分的相互作用及其对角质形成细胞和成纤维细胞的不同影响的发现。这项申请将扩展到以前的博士学位目前无法触及的领域，并在商业上应用。具体来说：使用太阳模拟器(作为主要光源)以及应用不同定制滤光片组合的详细方法，该项目将研究可见光、红外和紫外线(UVA和B)的各种带宽分量对细胞路径的影响(在存在Croda活性物质的情况下，见下文)，如使用RNAseq技术确定的并通过下游分子方法进一步研究的那样。凯瑟琳·波恩目前正在表演开创性的展会。RNA-Seq有助于观察替代基因剪接转录本、转录后修饰、基因融合、突变/SNPs和基因表达随时间的变化，或不同组或治疗组中基因表达的差异。除了mRNA转录本，RNA-Seq还可以查看总RNA、小RNA，如miRNA、tRNA和核糖体图谱、外显子/内含子边界。研究不同的商用防晒霜及其不同波长的透射率如何影响上述细胞通路的调制。确定UVR的单独成分(UVA和UVB)在不同可见光段的情况下的组合效果。这将涉及上述途径的生物标记物和DNA损伤(线粒体DNA、核)、氧化应激、ICM生物标记物的测量。研究不同类型的二氧化钛和氧化锌在相对过滤从UVR到红外的太阳光谱方面的保护情况，根据减少的UVR透射量是否足以影响可见光和红外线的生物反应，如以下FASE B论文所述。初步实验将使用原代人类皮肤细胞的单层培养，并逐步使用3D人类皮肤模型(比较LabSkin和Keratify的例子)。参考文献：哈德森·L、拉什丹·E、波恩·C、查万·B、罗林斯·D和白桦树-马赫。太阳辐射暴露的紫外线、红外和可见光成分对人体皮肤分子生物标志物的单一和综合影响.FASE B 2020 MAR；34(3)：3874-3883.

Skin ageing is a complex process that is determined by both intrinsic and extrinsic factors, which leads to a progressive loss of structure and function. Mitochondria are the major source of cellular oxidative stress and are widely implicated in cutaneous ageing as they have direct effects on the bioenergy of skin as well as affecting other cellular processes and nuclear DNA damage, which will also play a key role in skin ageing. Extrinsic skin ageing is driven to a large extent by environmental factors and external stressors such as UVR (and more recently visible and infrared components of sunlight), pollution and lifestyle factors have been shown to stimulate the production of reactive oxygen species and generate oxidative stress. The damage from these exogenous sources can impair skin structure and function leading to the phenotypic features of extrinsic skin ageing. Plan of Investigation The PhD proposal will apply some of the technology that the previous PhD with Croda (Catherine Bonn student has established utilising the finding of the interaction of the UV, Visible and IR components of sunlight and their differential effects of keratinocytes and fibroblasts as published in our FASEB paper (2020). The application will extend and commercially apply into areas that are beyond the current reach of the previous PhD. In detail:Using the solar simulator (as the primary light source) together with the detailed methodology of the application of different combinations of bespoke filters, the project will study the effects of the various bandwidth components of visible light, infrared and the UVR (both UVA and B) on cellular pathways (in the presence of Croda actives, see point below) as identified using RNAseq technology and further investigated by downstream molecular methodologies. Catherine Bonn is currently performing the pioneering expts. RNA-Seq facilitates the ability to look at alternative gene spliced transcripts, post-transcriptional modifications, gene fusion, mutations/SNPs and changes in gene expression over time, or differences in gene expression in different groups or treatments. In addition to mRNA transcripts, RNA-Seq can look at total RNA, small RNA, such as miRNA, tRNA, and ribosomal profiling, exon/intron boundaries. There is the opportunity for single cell sequencing.Investigate how different commercial sunscreens and their differential wavelength transmittance affect the modulation of the cellular pathways identified above.To determine the combinational effects of the separate components of UVR (UVA and UVB) with or without the different segments of visible light. This will involve measurement of biomarkers of pathways identified above and DNA damage (mtDNA, nuclear), oxidative stress, ICM biomarkers. Investigate the profiles of protection of differing types of titanium dioxide and zinc oxide in terms of relative filtering of the solar spectrum ranging from UVR to infrared in terms of whether the reduced amount of transmitted UVR is enough to affect the biological responses of visible and infrared as detailed in the FASEB paper belowInitial experiments will use monolayers cultures of primary human skin cells with a progression towards the use of 3D human skin models (comparing examples of Labskin and Keratify). Reference: Hudson L, Rashdan E, Bonn C, Chavan B, Rawlings D, and Birch-Machin MA. Single and combinational effects of ultraviolet, infrared and visible components of solar radiation exposure on molecular biomarkers in human skin .FASEB 2020 Mar; 34(3): 3874-3883.