Hydrogen Sulfide and Carbonyl Sulfide Delivery for Biological Applications

项目摘要 从血管生物到组织，硫化氢在人类健康中扮演着重要的角色 再生。为了推进对硫化物作用的研究，研究人员经常使用硫化氢供体直接调节硫化物 在实验期间的水平。尽管有这种广泛的效用，但关键的未得到满足的需求仍将在本 求婚。这项研究的长期目标是开发和部署COS/H2S捐赠者来调查和 推进活性硫物种与人类健康的多方面作用。这个项目的总体目标是 建议拓宽COS/H2S释放基序的平台，了解不同CA的异构体 影响COS向H2S的转化，并将COS/H2S供体应用于体内骨再生模型。这个 这些研究的基本原理是，提供COS/H2S的功能工具和更好地了解 不同的CA异构体对COS的水解活性将使COS/H2S释放的未来应用成为可能 针对存在关键CA异构体且已知硫化氢起保护作用的系统。这个 拟议的调查包括三个具体目标：(1)COS/H2S的扩展、提炼和应用 释放平台；(2)不同CA亚型对COS水解酶活性的研究；(3) COS/H_2S供体在骨再生中的应用本建议书建立在建立COS之前的工作基础上 释放的分子可以作为硫化氢的供体，因为COS通过以下途径快速酶促转化为硫化氢 碳酸酐酶(CA)。在目标1中，使用了新的化学方法来实现COS/H2S的输送，扩展了 可追踪的COS/H2S供体的动态范围和调色板，并提供放大的释放。在目标2中，水解物 单独的CA异构体对COS和模型硫酯/硫代酯底物的活性被研究 了解CA对COS水解率的异构体差异。在目标3中，开发的COS/H2S 供体被用来研究硫化氢在细胞和动物骨再生模型中的作用。这种方法是 创新，因为它提供了COS/H2S交付的新方法，满足了现场未满足的关键需求 首次揭示了CA异构体在COS到H2S转化过程中的不同活性。此外， 拟议的骨再生应用是创新的，因为它们不仅利用保护性的 在成骨过程中，硫化氢和CA的高局部活性的影响，但也利用了未充分利用的连接 与破骨细胞内钙离子的募集有关的硫化氢。这项拟议的研究具有重要意义，因为它提供了新的 直接解决该领域关键限制的方法，包括放大的释放系统，可跟踪 捐赠者和分析物替换方法。此外，拟议中的研究提供了第一个洞察力 COS到H2S转化的CA异构体差异，这将在相关的模型系统中加以利用 在人体健康中，CA活性和硫化氢的释放都很重要。圆满完成拟议中的 AIMS将在硫化氢生物学领域产生积极影响，并将导致对这些角色的更好理解 COS/H2S对人体健康的影响。

Project Summary Hydrogen sulfide (H2S) plays important roles in human health ranging from vascular biology to tissue regeneration. To advance investigations into roles, researchers often use H2S donors to directly modulate sulfide levels during experiments. Despite this broad utility, key unmet needs remain that will be addressed in this proposal. The long-term goal of this research is to develop and deploy COS/H2S donors to investigate and advance the multifaceted roles of reactive sulfur species related to human health. The overall objectives of this proposal are to broaden the platform of COS/H2S releasing motifs, to understand how different CA isoforms impact COS to H2S conversion, and to apply COS/H2S donors to in vivo models of bone regeneration. The rationale for these studies is that the functional tools for COS/H2S delivery and a greater understanding of differential CA isoform activity toward COS hydrolysis will enable future applications in which COS/H2S release is targeted to systems where key CA isoforms are present and H2S is known to play a protective effect. The proposed investigations include three specific aims: (1) Expansion, refinement, and application of COS/H2S releasing platforms; (2) Investigation in to the differential CA isoform activity toward COS hydrolysis; and (3) Application of COS/H2S donors to bone regeneration. This proposal builds from prior work establishing that COS releasing molecules can function as H2S donors due to the rapid enzymatic conversion of COS to H2S by carbonic anhydrase (CA). In Aim 1, new chemical approaches are used to enable COS/H2S delivery, expand the dynamic range and palette of traceable COS/H2S donors, and provide amplified release. In Aim 2, the hydrolytic activity of individual CA isoforms toward COS and model thioester/thionoester substrates are investigated to understand isoform differences in CA efficiency toward COS hydrolysis. In Aim 3, the developed COS/H2S donors are used to investigate the role of H2S in cell and animal models of bone regeneration. This approach is innovative because it provides new approaches to COS/H2S delivery that address key unmet needs in the field and provides the first insights into the differential activity of CA isoforms for COS to H2S conversion. Moreover, the proposed applications in bone regeneration are innovative because they not only leverage the protective effects of H2S and high local activity of CA during osteogenesis, but also leverages the underutilized connection of H2S with Ca2+ recruitment in osteoclasts. The proposed research is significant because it provides new approaches that directly address key limitations in the field, including amplified release systems, trackable donors, and analyte replacement methodologies. In addition, the proposed research provides the first insights into CA isoform differences for COS to H2S conversion, which will be leveraged in model systems relevant to human health in which both CA activity and H2S delivery are important. Successful completion of the proposed Aims will provide a positive impact in the field of H2S biology and will result a greater understanding of the roles of COS/H2S in human health.