A Chemical Toolkit to Define Formaldehyde’s Enigmatic Biology

甲醛（HCHO）是最简单的醛，是一种高活性的人体代谢物。尽管它在高浓度下具有毒性和致癌性，并在范可尼贫血患者中具有诱导疾病的作用，但其生物学机制却非常未知。最近通过染色质去甲基化酶产生HCHO的开创性发现，以及最近对HCHO介导的C1代谢的研究，表明HCHO在“正常”条件下的功能相关作用。然而，由于缺乏方法，这方面的详细研究一直受到阻碍。从化学的角度来看，HCHO与生物分子有很强的反应性。这种反应性被用于防腐和免疫沉淀测定，可能是其生物学功能的原因。然而，缺乏对化学的了解。鉴于细胞中HCHO的常规生产及其高浓度毒性，其调节人类健康的潜力是明确而引人注目的。因此，我假设内源性HCHO是人类生物学中关键的功能调节剂，并且可能是人类细胞中最重要的小分子之一。细胞HCHO水平失调可能导致疾病，使HCHO成为潜在的“沉默杀手”。用小分子调节细胞HCHO也可能具有治疗潜力，为药物发现开辟了新的途径。HCHO的反应性、挥发性和体积小，使其在生物系统中的操作和研究非常具有挑战性。因此，HCHO的生物学特性在很大程度上被忽视了。因此，在这个项目中，我将利用我在有机合成、生物化学、细胞生物学和分析技术方面的多学科背景来开发第一个“化学HCHO工具包”——这个工具包将使HCHO在人类生物学中的功能首次得到系统的定量分析。具体目标是：(1)合成和验证一流的HCHO释放小分子（FORMs），使HCHO在细胞中可控制的可量化释放成为可能。(2)合成并验证可降低细胞HCHO浓度的不可逆HCHO清除剂（FOSCs）。(3)开发并验证了第一个使用不可逆FOSCs与NMR和GC/MS分析相结合的细胞HCHO定量方法。这个开创性的工具包将使识别HCHO和细胞分子之间的反应成为可能，以确定HCHO水平在健康和疾病中如何变化，将这些变化与生物表型相关联，并最终验证HCHO调节作为一种治疗策略。这些能力目前还不可能实现，但对于研究细胞中的HCHO是必不可少的。因此，这个雄心勃勃的、及时的项目将为在英国和国际上建立和推进HCHO生物研究提供最先进的平台技术。这项工作还将使我的团队处于这一潜在变革领域的前沿，并最终导致人类疾病的新疗法。

Formaldehyde (HCHO), the simplest aldehyde, is a highly reactive human metabolite. Despite its toxicity and carcinogenicity at high concentrations and its disease-inducing effects in Fanconi's anaemia patients, the mechanisms underpinning its biology are remarkably unknown. The recent pioneering discovery of HCHO production by chromatin demethylases, as well as recent work on HCHO-mediated C1 metabolism, suggest functionally relevant roles for HCHO under 'normal' conditions. However, detailed research in this area has been precluded by a lack of methods.From a chemical perspective, HCHO is highly reactive with biomolecules. This reactivity, which is exploited for uses e.g. in embalming and immunoprecipitation assays, is likely responsible for its biological functions. However, chemical insight is lacking.Given the routine production of HCHO in cells and its toxicity at high concentrations, the potential for it to regulate human health is clear and compelling. I therefore hypothesise that endogenous HCHO is a key functional regulator in human biology and potentially one of the most important small molecules in human cells. Dysregulation of cellular HCHO levels is likely to cause disease, making HCHO a potential 'silent killer'. Modulating cellular HCHO with small molecules is also likely to have therapeutic potential, opening up new avenues for drug discovery.HCHO's reactivity, volatility and small size make it very challenging to manipulate and study in biological systems. Consequently, HCHO's biology has been largely overlooked. In this project, I will therefore use my multidisciplinary background in organic synthesis, biochemistry, cell biology and analytical techniques to develop the first 'chemical HCHO toolkit' - this toolkit will enable the first systematic quantitative analysis of HCHO's functions in human biology. Specific objectives are:(1) To synthesise and validate first-in-class HCHO-releasing small molecules (FORMs) that enable controlled quantifiable release of HCHO in cells.(2) To synthesise and validate irreversible HCHO scavengers (FOSCs) that can reduce cellular HCHO concentrations.(3) To develop and validate the first sensitive cellular HCHO quantification methods using irreversible FOSCs coupled to NMR and GC/MS analyses.This pioneering toolkit will make it possible to identify reactions between HCHO and molecules in cells, to determine how HCHO levels vary in health and disease, to correlate these changes with biological phenotypes, and ultimately to validate HCHO modulation as a therapeutic strategy. These abilities are currently not possible but are essential to studying HCHO in cells. Therefore, this ambitious and timely project will provide the state-of-the-art platform technologies essential to establishing and furthering HCHO biological research in the UK and internationally. The work will also position my group at the forefront of this potentially transformative field, and will ultimately lead to new treatments for human diseases.