Developing Pharmacological tGLI1 Inhibitors

开发药理学 tGLI1 抑制剂

• 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
• 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

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 乳腺癌脑转移（BCBM）患者在诊断后仅存活6-18个月。它仍然是一个 在管理BCBM患者方面存在重大挑战。因此，本项目的目标是满足识别 新的可操作的目标和开发针对BCBM的这些目标的治疗方法。根据我们初步的 结果总结如下，我们将集中在截短的胶质瘤相关癌基因同源物1 （tGLI 1），在申办方/导师实验室发现，是一种肿瘤特异性功能获得性GLI 1锌指 促进乳腺癌优先转移到大脑的转录因子，乳腺癌干细胞 （BrCSC）和脑转移灶中的转移支持星形胶质细胞。在初步研究中，我们 筛选了1,520种化合物，并选择性地鉴定了酮康唑（KCZ），一种FDA批准的唑类抗真菌剂， 在体外杀死表达tGLI 1的乳腺癌细胞和BCBM细胞，提高对BrCSC的效力。 两项心内植入小鼠研究显示，全身KCZ给药选择性地阻止了 循环中tGLI 1阳性乳腺癌细胞发展成脑转移，并抑制 现有脑转移的进展。我们进一步表明KCZ减少了tGLI 1介导的 体内循环外泌体microRNA，miR-1246和miR-1290，KCZ有效地渗透血液， 脑屏障（BBB），对肝组织无急性毒性。虽然KCZ抑制的作用机制， tGLI 1目前还不清楚，我们的初步数据表明KCZ可以抑制tGLI 1的转录活性并破坏tGLI 1 与细胞蛋白质的相互作用。这些重要的新发现导致了FDA批准的0期临床试验。 维克森林综合癌症中心的一项试验，确定KCZ对tGLI 1靶基因的影响 表达和KCZ渗透的BCBM患者的血脑屏障。基于这些观察，我们假设 KCZ在抑制BCBM中的tGLI 1靶基因方面具有临床活性，并且新型tGLI 1抑制剂可以 可以通过KCZ侧基的修饰或片段化的命中化合物的SAR分析来开发。到 为了验证这一创新假设，我们将测试KCZ衍生物对tGLI 1阳性BCBM的作用，以确定它们是否 使用细胞系和PDX衍生的BCBM， 小鼠模型，并阐明其活性的机制（目的1），并测试从头tGLI 1 通过KCZ的构效关系（SAR）分析合成抑制剂WF-217 A和WF-229 A 碎片（目标2）。这个F31博士前奖学金是量身定制的，以促进申请人的发展成为一个 进行转化性癌症研究的独立调查员。这一目标将通过相关的 课程工作，每周研讨会和期刊俱乐部介绍，在国家会议上介绍，赠款-和 编写文件讲习班，与主要导师和辅导小组定期举行会议（4名导师， 相关专业知识），以及使用临床前模型测试创新假设的研究培训。