High Throughput Screening (HTS) to Discover Graft-Versus-Host Disease Inhibitors

高通量筛选 (HTS) 发现移植物抗宿主疾病抑制剂

基本信息

批准号：
8474927
负责人：
Sophie Paczesny
金额：
$ 33.76万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-08 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8474927
关键词：
Acute Graft Versus Host Disease Affect Allogenic Asthma Automobile Driving Binding Binding Sites Biological Biological Assay Biological Factors Biological Markers Biological Models Blood Chemicals Clinic Clinical Collection Complication Computer Simulation Data Detection Development Disease Docking Drug Targeting Effector Cell Enzyme-Linked Immunosorbent Assay Equilibrium Evaluation Exhibits Foundations Functional disorder Goals Heart failure Hematological Disease Hematopoietic Stem Cell Transplantation Human Immune response Immunotherapeutic agent In Vitro Lead Libraries Life Ligands Malignant - descriptor Malignant Neoplasms Mediating Modeling Mus Outcome Pathology Pathway interactions Patients Pharmaceutical Preparations Phenotype Plasma Play Pre-Clinical Model Property Proteins Publishing Research Role Signal Transduction Steroid therapy Steroids Structure Symptoms T-Lymphocyte Technology Testing Th2 Cells Therapeutic Therapeutic Intervention Toxic effect Tumorigenicity United States National Institutes of Health Validation Work base cancer therapy extracellular graft vs host disease high throughput screening in vitro Model in vivo inhibitor/antagonist innovation insight molecular dynamics mortality new therapeutic target novel overexpression pharmacophore public health relevance receptor response simulation small molecule small molecule libraries tandem mass spectrometry therapeutic development therapeutic evaluation therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Allogeneic hematopoietic stem cell transplantation (HSCT) is a potentially curative therapy for many malignant diseases but its clinical utility has been impeded by graft-versus-host disease (GVHD). Unfortunately, there are currently no therapies that target proteins that are elevated in the blood of patients presenting with GVHD, and no effort has been made to develop drugs specific for GVHD. This represents an important problem, because currently therapies are limited to the nonspecific targeting of effector cells. Our long-term goal is to develop drugs targeting proteins that are dysregulated in patients with GVHD as potential novel GVHD therapies. Our objective in this application is to discover chemical probes that can inhibit these elevated proteins through high throughput screening (HTS) of small molecules libraries and be evaluated in the biological model systems. These compounds will allow for discovery of potential GVHD-specific drugs and therefore better harnessing of treatment in many patients with hematological cancers. Our central hypothesis is that the lead plasma biomarker of GVHD, suppression of tumorigenicity 2 (ST2) can be targeted with small molecule compounds to alleviate GVHD. This hypothesis was formed based on our published data characterizing the soluble form of ST2 (sST2) as the most significant biomarker to predict the non-response to GVHD therapy. sST2 acts as a decoy receptor for IL-33, the only known ligand for ST2, driving Th2 cells toward a Th1 phenotype. This phenotypic change leads to GVHD, and an inhibitor of the binding association between sST2 and IL-33 could alleviate GVHD. We therefore embark on a pilot HTS campaign using the AlphaScreen detection assay to discover chemical probes that inhibit the ST2 and IL33 interaction. Our optimized assay condition was robust giving z'-factors e 0.7 and permitted discovery of 17 hits. The rationale for this study is that the likelihood for better probes than those in the pilot screen warrants a large screen. In addition, a platform including in silico enrichment of potential hits from broader compound libraries and testing of the chemical probes in biological models of GVHD will be relevant. The three specific aims are: 1) HTS implementation to discover chemical probes that inhibit the ST2/IL33 pathway that is overexpressed in graft-versus-host disease. 2) Enrich hits using focus compound libraries generated from pharmacophore models based on confirmed hits from HTS and identification of putative binding sites in ST2. 3) Hits validation with in vitro huma assays and in vivo murine models of GVHD. This integrative approach is innovative because it applies HTS technology of 176,000 diverse small molecules with in silico enrichment of hits and target binding site evaluation yielding the identification and development of chemical probes against biomarkers of GVHD refractoriness to therapy. The proposed research is significant because discovery of GVHD-specific inhibitors will provide the foundation for therapeutic evaluation of GVHD biomarkers as druggable targets. The application of these drugs will drive the phenotypic changes in appropriate effector T cells with increased efficacy and lowered toxicity.

描述（由申请人提供）：同种异体造血干细胞移植（HSCT）是许多恶性疾病的潜在治疗疗法，但其临床实用性受到了临床疗法的影响，受到了疗法 - 抗抗菌病（GVHD）。不幸的是，目前尚无靶向蛋白质升高的蛋白质升高的患者的血液中的蛋白质，也没有努力开发针对GVHD的药物。这代表了一个重要的问题，因为目前的疗法仅限于效应细胞的非特异性靶向。我们的长期目标是开发针对GVHD患者作为潜在的GVHD疗法的患者失调的蛋白质的药物。我们在本应用中的目标是发现可以通过小分子文库的高吞吐量筛选（HTS）抑制这些升高蛋白的化学探针，并在生物模型系统中进行评估。这些化合物将允许发现潜在的GVHD特异性药物，因此可以更好地利用许多血液学癌症患者的治疗。我们的中心假设是GVHD的铅血浆生物标志物，抑制肿瘤性2（ST2）可以用小分子化合物靶向以减轻GVHD。该假设是根据我们发布的数据来形成的，该数据表征了ST2（SST2）的可溶形式，是预测GVHD疗法无反应的最重要的生物标志物。 SST2充当IL-33的诱饵受体，IL-33是唯一已知的ST2配体，将Th2细胞驱动到Th1表型。这种表型变化导致GVHD，SST2和IL-33之间结合关联的抑制剂可以减轻GVHD。因此，我们使用Alphascreen检测测定法进行了试点HTS运动，以发现抑制ST2和IL33相互作用的化学探针。我们优化的测定条件是可靠的，从而使Z'Factors E 0.7并允许发现17次命中。这项研究的理由是，比飞行员屏幕中的探针更好的探针可能是大屏幕。此外，在GVHD生物学模型中，包括从更宽的化合物库中的硅富集和化学探针测试的平台将是相关的。三个具体目的是：1）HTS实施，以发现抑制在移植物与宿主病中过表达的ST2/IL33途径的化学探针。 2）使用基于HTS的确认命中和ST2中假定结合位点的确认命中产生的焦点化合物文库丰富命中。 3）通过体外Huma分析和GVHD的体内鼠模型进行验证。这种综合方法具有创新性，因为它采用了176,000个不同小分子的HTS技术，并在击中的硅富集中和目标结合位点评估中，可鉴定和开发针对GVHD冰淇淋性生物标志物的化学探测器对治疗的生物标志物。拟议的研究很重要，因为发现GVHD特异性抑制剂将为GVHD生物标志物作为可药物靶标提供治疗评估的基础。这些药物的应用将驱动适当效应T细胞的表型变化，并增加毒性并降低毒性。