Smooth muscle cell-derived cell fates and cellular interactions in atherosclerotic plaque stability in disease progression and regression.

接受降脂治疗的患者中的残余心血管疾病（CVD）风险是一个主要的未满足的临床需求。 我们的目标是研究斑块稳定和不稳定的机制，重点是血管功能， 小鼠模型中平滑肌细胞（SMC）衍生细胞（SDC）及其与巨噬细胞（M β）的相互作用 和人类CVD。我们重点关注SDC的类型、位置和功能及其在斑块稳定性中与M β的相互作用 疾病的进展和消退。SMC可以通过中间状态转变为动脉粥样硬化保护， 例如，在一个实施例中，纤维软骨细胞（SMC-FbC），或致动脉粥样硬化，例如，SMC衍生的巨噬细胞样（SMC-M）身份。 SDC身份的主要调节剂正在出现保护性（视黄酸，Tcf 21）和有害（Klf 4）类型， 但机制却知之甚少。我们假设SDC在病变稳定性和临床CVD中起作用， 是可塑性的，可以从空间和单细胞（sc）组学推断; SMC-M是炎症性的，激活 旁观者M β促进斑块不稳定性;氧化DNA（ox-DNA）损伤调节SDC身份， 功能使病变不稳定;致动脉粥样硬化的SDC和M β的串扰促进病变不稳定， 会影响回归目的1明确SDC类型在动脉粥样硬化进展中的定位和功能 并测试SMC-M β和SDC ox-DNA损伤是否驱动致动脉粥样硬化的SMC特性和病变不稳定性，而Aim 2将解决在动脉粥样硬化消退中，一些SDC是否促进斑块稳定，而另一些SDC是否抑制斑块稳定， SMC-M β和SDC ox-DNA损伤是否减弱了消退。在这些目标中，我们将：（1）整合 基于空间杂交的RNA原位测序（HybRISS）、sc-omics和SMC谱系追踪， 病变进展和消退期间SDC的身份、空间功能和主调节因子，重点关注 功能，如SMC-M炎性小体和受损的白细胞增多症;（2）使用白喉毒素（Dtx）小鼠 跟踪SMC-M β的模型，并测试SMC-M β的耗竭是否减少进展中的病变不稳定性， 加速分解;和（3）使用SMC诱导的8-氧代鸟嘌呤DNA糖基化酶（OGG 1）转基因小鼠 模型，以测试在动脉粥样硬化中增加的SMC ox-DNA损伤是否将SDC推向动脉粥样硬化 身份目的3将检测特定的SDC类型及其调控基因是否促进临床CVD。使用多组学 基于来自慕尼黑血管生物库的数据，我们将部署一项综合策略，从描述性研究， 人类斑块，SDC中的调节性遗传变异，测试这些变异与 临床CVD在大型遗传数据。我们的工作将提供对SDC类型的作用的机械见解， 空间功能，以及与M的串扰，以实现我们的总体目标，即更好地理解 影响动脉粥样硬化斑块稳定性和不稳定性的因素。人工翻译将建立临床相关性， CVD新治疗策略的因果关系和背景。

Residual cardiovascular disease (CVD) risk in patients on lipid lowering therapy is a major unmet clinical need. Our goal is to investigate mechanisms of plaque stabilization and destabilization focused on functions of vascular smooth muscle cell (SMC) derived cells (SDCs), and their crosstalk with macrophages (Mϕ), in mouse models and human CVD. We focus on SDC types, location and functions and their interactions with Mϕ in plaque stability in disease progression and regression. SMCs can transition through an intermediate state into atheroprotective, e.g., fibrochondrocyte (SMC-FbC), or atherogenic, e.g., SMC-derived macrophage-like (SMC-Mϕ) identities. Master regulators of SDC identities are emerging for protective (retinoic acid, Tcf21) and harmful (Klf4) types, but mechanisms are poorly understood. We hypothesize that SDC functions in lesion stability and clinical CVD are malleable and can be inferred from spatial and single-cell (sc) omics; SMC-Mϕ are inflammatory and activate bystander Mϕ to promote plaque instability; oxidative DNA (ox-DNA) damage regulates SDC identities and functions to destabilize lesions; and crosstalk of atherogenic SDCs and Mϕs promotes lesion instability and impairs regression. Aim 1 will determine locations and functions of SDC types in atherosclerosis progression and test if SMC-Mϕ and SDC ox-DNA damage drive atherogenic SMC identities and lesion instability while Aim 2 will address whether some SDCs promote and others curb plaque stabilization in atherosclerosis regression, and if regression is attenuated by SMC-Mϕ and SDC ox-DNA damage. In these Aims, we will; (1) integrate spatial hybridization-based RNA in situ sequencing (HybRISS), sc-omics and SMC lineage tracing to define identity, spatial functions, and master regulators of SDCs during lesion progression and regression, focusing on functions such as SMC-Mϕs inflammasome and impaired efferocytosis; (2) use a diphtheria toxin (Dtx) mouse model to track SMC-Mϕ and to test if depletion of SMC-Mϕ reduces lesion instability in progression and accelerates resolution; and (3) use an SMC-inducible 8-oxoguanine DNA glycosylase (OGG1) transgenic mouse model to test if SMC ox-DNA damage, which is increased in atherosclerosis, pushes SDCs to atherogenic identities. Aim 3 will test if specific SDC types and their regulatory genes promote clinical CVD. Using multi-omic data from the Munich Vascular Biobank, we will deploy an integrated strategy to go from descriptive studies in human plaques, to regulatory genetic variation in SDCs, to testing causal relationship of these variants with clinical CVD in large genetic data. Our work will provide mechanistic insight into the role of SDC types, their spatial functions, and crosstalk with Mϕ to achieve our overall goal of greater mechanistic understanding of factors affecting atherosclerotic plaque stability and instability. Human translation will establish clinical relevance, causality and context for new treatment strategies for CVD.