High Dimensional Mass Cytometry Analysis of the Effects of Vedolizumab in Intestinal Cellular Subsets and its Correlation with Clinical Parameters

项目摘要 我们对被vedolizumab和其他有效药物消耗的特定细胞亚群的理解 肠道内的药物仍然是表面的。这在一定程度上是由于技术限制，这些限制阻碍了 对人体肠道固有层细胞和分子组成的无偏见询问。 最近，这些技术挑战已经被质量细胞术的发展所克服，它 可同时定量多达40个细胞亚群的表面和/或细胞内标记。我们 最近采用了以前主要用于外周血的现有质量细胞分析方法 细胞分析，以允许询问肠道固有层细胞。我们已经开发、测试和 优化了一组37个免疫细胞标志物，使我们能够同时识别所有主要的免疫细胞 以及许多亚群、细胞内细胞因子和转录因子。在这里我们 是否会通过深部手术来前瞻性地评估vedolizumab对肠道细胞成分的影响 给药前后免疫细胞亚群的免疫表型。到时候我们会的 确定细胞亚群的变化是否与临床、内窥镜或组织学参数相关 疾病活动。这些研究将检验质量细胞术结果作为人类免疫缺陷的替代标志物的有效性。 靶向淋巴细胞运输的药物(vedolizumab、etrolizumab、抗MAdCAM-1、奥扎尼莫德)和 可能直接阐明这些交通靶向疗法的具体作用机制。

Project Summary Our understanding of the specific cell subsets that are being depleted by vedolizumab and other effective agents within the intestine remains superficial. This is in part due to technical limitations which had hampered the unbiased interrogation of cellular and molecular composition of the human intestinal lamina propria. Recently, these technological challenges have been overcome by the development of mass cytometry, which enables the simultaneous quantification of up to 40 surface and/or intracellular markers on cell subsets. We have recently adapted existing mass cytometry methodology previously employed mainly for peripheral blood cell analyses, to allow for the interrogation of intestinal lamina propria cells. We have developed, tested and optimized a panel of 37 immune cell markers that allow us to simultaneously identify all major immune cell populations, as well as numerous sub-populations, intracellular cytokines and transcription factors. Here we will prospectively assess the effects of vedolizumab on the cellular composition of intestine by performing deep immunophenotyping of immune cell subsets, prior to and after the drug’s administration. We will then determine whether changes on cell subsets correlate with clinical, endoscopic or histologic parameters of disease activity. These studies will test the validity of mass cytometry outcomes as surrogate markers for the efficacy of drugs that target lymphocyte traffic (vedolizumab, etrolizumab, anti- MAdCAM-1, ozanimod) and may directly shed light on the specific mechanism of action of these traffic-targeted therapies.