Developing Pharmacological tGLI1 Inhibitors

开发药理学 tGLI1 抑制剂

基本信息

批准号：
10388709
负责人：
Daniel Doheny
金额：
$ 2.9万
依托单位：
WAKE FOREST UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-20 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10388709
关键词：
Alternative Splicing Amino Acids Antifungal Agents Astrocytes Azoles Blood - brain barrier anatomy Brain Breast Cancer Cell CD44 gene Cancer Patient Cell Line Cells Clinical Comprehensive Cancer Center Data Development Diagnosis Down-Regulation Educational workshop Excision Exons FDA approved Fellowship GLI gene Gene Expression Genes Genetic Transcription Glioblastoma Goals Grant Growth In Vitro Internal Breast Prosthesis Journals KDR gene Ketoconazole Libraries Liver Manuscripts Mediating Mentors Metastatic malignant neoplasm to brain MicroRNAs Modification Molecular Mus Neoplasm Metastasis Normal Cell Nuclear Localization Signal Oncogenic Operative Surgical Procedures Pathologic Patients Penetrance Pharmaceutical Preparations Pharmacology Phase 0 Clinical Trial Play Pre-Clinical Model Process Proteins Publishing Radiation therapy Radiosurgery Recurrence Research Personnel Research Training Role STAT3 gene Sampling Side Specificity Structure-Activity Relationship Testing Tissues Toxic effect United States Variant Vascular Endothelial Growth Factor C Vascular Endothelial Growth Factors Woman Work Writing Zinc Fingers acute toxicity angiogenesis base blood-brain barrier penetration blood-brain tumor barrier cancer stem cell design forest gain of function implantation improved in vivo inhibitor innovation knock-down malignant breast neoplasm meetings migration mouse model novel patient derived xenograft model pre-doctoral prevent prognostic indicator standard of care symposium targeted agent therapeutic target transcription factor translational cancer research tumor

项目摘要

Project Summary Patients with breast cancer brain metastasis (BCBM) survive only 6-18 months following diagnosis. It remains a significant challenge in managing BCBM patients. The goal of this project is thus to meet the need of identifying novel actionable targets and developing treatments against these targets for BCBM. Building on our preliminary results summarized below, we will focus the project on truncated glioma-associated oncogene homolog 1 (tGLI1), which was discovered in Sponsor’s/Mentor’s Lab as a tumor-specific gain-of-function GLI1 zinc-finger transcription factor that promotes breast cancer preferential metastasis to the brain, breast cancer stem cells (BrCSCs), and metastasis-supporting astrocytes in the brain metastatic niche. In the preliminary studies, we screened 1,520 compounds and identified ketoconazole (KCZ), an FDA-approved azole antifungal, selectively killed tGLI1-expressing breast cancer cells and BCBM cells with heightened efficacy against BrCSCs in vitro. Two intracardiac implantation mouse studies showed systemic KCZ administration selectively prevented circulating tGLI1-positive breast cancer cells from developing into brain metastases and suppressed the progression of existing brain metastases. We further showed KCZ reduced tGLI1-mediated expression of circulating exosomal microRNAs, miR-1246 and miR-1290, in vivo, and KCZ effectively penetrated the blood- brain barrier (BBB) without acute toxicity to liver tissues. While the mechanisms of action for KCZ inhibition of tGLI1 are still unclear, our pilot data suggest that KCZ can inhibit tGLI1 transcriptional activity and disrupt tGLI1 interactions with cellular proteins. These important novel findings have led to an FDA-approved Phase 0 clinical trial at Wake Forest Comprehensive Cancer Center that determines the effects of KCZ on tGLI1 target gene expression and KCZ penetration of the BBB in BCBM patients. Based on these observations, we hypothesized that KCZ elicits clinical activity in suppressing tGLI1 target genes in BCBM and that novel tGLI1 inhibitors can be developed through either modification of KCZ side groups or SAR analysis of fragmented hit compounds. To test this innovative hypothesis, we will test KCZ derivatives against tGLI1-positive BCBM to determine if they present with increased efficacy and retain tGLI1-targeting specificity using cell line- and PDX-derived BCBM mouse models, and elucidate the mechanisms that underlie their activity (Aim 1), and test the de novo tGLI1 inhibitors WF-217A and WF-229A synthesized via structure-activity relationship (SAR) analysis of KCZ fragments (Aim 2). This F31 predoctoral fellowship is tailored to facilitate the applicant’s development into an independent investigator who conducts translational cancer research. This goal will be achieved through relevant course work, weekly seminars and journal club presentations, presentation at national conferences, grant- and manuscript-writing workshops, regular meetings with the primary mentor and the mentoring team (4 mentors with relevant expertise), and research training that tests an innovative hypothesis using preclinical models.

项目摘要诊断后仅6-18个月，患有乳腺癌脑转移（BCBM）的患者生存。它仍然是一个管理BCBM患者的重大挑战。因此，该项目的目的是满足确定的需求新颖的可行靶标并开发针对BCBM的这些靶标的治疗方法。基于我们的初步结果总结了下面的结果，我们将将该项目集中在截短的神经胶质瘤相关的癌基因同源物1 （TGLI1），在赞助商/导师的实验室中发现是肿瘤特异性的GLI1锌指促进乳腺癌优先转移大脑的转录因子，乳腺癌干细胞（BRCSC），以及脑转移性小众中支持转移的星形胶质细胞。在初步研究中，我们筛选了1,520种化合物，并鉴定出酮康唑（KCZ），一种由FDA批准的azole抗真菌剂，有选择性地在体外杀死了表达TGLI1的乳腺癌细胞和BCBM细胞的效率。两项心脏内植入小鼠研究表明，全身性KCZ有选择地预防循环TGLI1阳性乳腺癌细胞从发展为脑转移并抑制现有脑转移的进展。我们进一步显示KCZ降低了TGLI1介导的表达循环的外泌体microRNA，miR-1246和miR-1290，体内和KCZ有效地穿透了血液脑屏障（BBB）对肝组织没有急性毒性。而KCZ抑制的作用机制 TGLI1仍不清楚，我们的试点数据表明，KCZ可以抑制TGLI1转录活性并破坏TGLI1 与细胞蛋白的相互作用。这些重要的新颖发现导致了FDA批准的0阶段临床 Wake Forest综合癌症中心的试验确定KCZ对TGLI1靶基因的影响 BCBM患者中BBB的表达和KCZ渗透。基于这些观察，我们假设 KCZ在抑制BCBM中抑制TGLI1靶基因的临床活性，而新型的TGLI1抑制剂可以可以通过修改KCZ侧组或碎片命中化合物的SAR分析来开发。到检验这种创新的假设，我们将针对TGLI1阳性BCBM测试KCZ衍生物，以确定它们是否是否使用细胞系和PDX衍生的BCBM提高效率并保留TGLI1靶向特异性鼠标模型，并阐明其活性的基础的机制（AIM 1），并测试Novo Tgli1 抑制剂WF-217A和WF-229A通过KCZ的结构活性关系（SAR）分析合成碎片（目标2）。这项F31餐饮前奖学金是为了促进申请人的发展而定制的进行翻译癌症研究的独立研究者。这个目标将通过相关实现课程工作，每周的半手和期刊俱乐部演讲，在国家会议上的演讲，授予和手稿编写讲习班，与主要导师和指导团队的定期会议（4位导师与相关专家）以及使用临床前模型检验创新假设的研究培训。