Identifying Neutrophil Specific Mechanisms for Resistance to Biologics in Ulcerative Colitis

摘要 溃疡性结肠炎(UC)是一种慢性炎症性肠病，在美国约有100万人患病。 美国。加州大学每年的医疗支出估计超过100亿美元，其中大多数 这笔费用与昂贵的生物疗法有关。UC中处方最多的两种生物制品是 然而，vedolizumab和adalimumab对这些高级疗法的有效率约为30%。 因此，了解治疗耐药的药物特异性机制是一个尚未得到满足的需求。我们的参与者 水平的患者数据和在小鼠结肠炎/结肠损伤模型中的观察表明，不同的 上皮内间隙的中性粒细胞亚群导致持续的肠道干细胞(ISC)损伤，阴性 生物制剂对上皮隐窝微环境和治疗效果的影响。因此，拟议的研究将 整合以患者为中心的临床研究和以基本机制为中心的研究，探索空间上截然不同的想法 具有独特遗传特征和功能的中性粒细胞群体可能决定特定的治疗机制 反应和影响粘膜损伤的解决。 在第一个目标中，使用空间分辨的单细胞和亚细胞高复合体数字定量的mRNA和 在已经收集的UC患者的粘膜组织中，我们将分析中性粒细胞、T细胞和肠道 UC患者基于空间方位的上皮(隐窝与固有层)。使用这种方法，我们将 确定预测UC患者对vedolizumab治疗反应的基因/蛋白质特征，并确定有针对性的 上皮性中性粒细胞与隐窝和T细胞相互作用的机制 Vedolizumab。第二个目标是研究上皮中性粒细胞负荷如何影响治疗 小鼠结肠炎对结肠损伤的反应和解决。具体地说，使用小鼠结肠损伤/结肠炎模型， 临床相关的治疗学和中性粒细胞特异的遗传方法，我们将定义分子线索指导 中性粒细胞在上皮内间隙的滞留以及这种独特的空间定位如何影响治疗 对生物制品(vedolizumab、adalimumab和tofacitinib)的反应。我们将进一步确定内部是否 上皮性中性粒细胞通过增强Vedolizumab的作用加重结肠损伤并阻碍其疗效 ISC损失。 因此，目前提案中概述的研究将确定治疗耐药的新机制 生物制剂和识别分子靶标以优化/提高响应性。

Abstract Ulcerative colitis (UC) is a chronic inflammatory bowel disease affecting approximately 1 million people in the United States. Annual healthcare expenditure for UC is estimated to be more than $10 billion dollars, with most of this cost being related to expensive biologic therapies. The two most prescribed biologics in UC are vedolizumab and adalimumab, however, efficacy for these advanced therapies plateaus at approximately 30%. Thus, there is an unmet need to understand drug specific mechanisms of treatment resistance. Our participant level patient data and observations in murine colitis/colon injury models indicate that persistence of distinct neutrophil subsets in the intra-epithelial space results in ongoing intestinal stem cell (ISC) injury, negatively impacting the epithelial crypt microenvironment and treatment efficacy for biologics. Thus, proposed studies will integrate clinical patient-focused and basic mechanism-focused studies to explore the idea that spatially distinct neutrophil populations with unique genetic signatures and function may dictate mechanism specific therapeutic response and impact mucosal injury resolution. In the first Aim using spatially resolved single-cell and sub-cellular high-plex digital quantification of mRNA and protein in already collected mucosal tissue from UC patients, we will profile neutrophils, T-cells, and intestinal epithelium based on spatial orientation (crypts vs lamina propria) in UC patients. Using this approach, we will identify gene/protein signatures predictive of response to vedolizumab therapy in UC and define targetable mechanisms through which epithelial neutrophils interact with crypts and T-cells to mediate non-response to vedolizumab. Second Aim will investigate how intra-epithelial neutrophil burden functionally impacts therapeutic responses and resolution of colon injury in murine colitis. Specifically, using murine colon injury/colitis models, clinically relevant therapeutics and neutrophil-specific genetic approaches, we will define molecular cues guiding neutrophil retention in the intra-epithelial space and how this unique spatial localization impacts therapeutic response to biologics (vedolizumab, adalimumab and tofacitinib). We will further determine whether intra- epithelial neutrophils exacerbate colon injury and impede therapeutic efficacy of vedolizumab through enhanced ISC loss. As such, studies outlined in the current proposal will identify new mechanisms of therapeutic resistance to biologics and identify molecular targets to optimize/improve responsiveness.