Identification of TR4 Modulators for Treatment of Cushing Disease.

用于治疗库欣病的 TR4 调节剂的鉴定。

• 批准号: 10199436
• 负责人: ANTHONY P HEANEY
• 金额: $ 40.11万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10199436
• 关键词: Adrenal Glands; Adrenalectomy; Affect; Bilateral; Biochemical; Biological Assay; Cells; Chemicals; Clinical Research; Clinical Trials; Corticotropin; Databases; Disease; Dose; Elements; Evaluation; Excision; Future; Genetic Transcription; Glucocorticoids; Goals; Health Care Costs; Homeostasis; Hormone secretion; Human; Hybrids; Hydrocortisone; Immunoassay; In Vitro; Lead; Length; Libraries; Life; Ligand Binding Domain; Luciferases; Medical; Morbidity - disease rate; Mus; Nuclear Orphan Receptor; Orphan; POMC gene; Patients; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Pituitary Corticotropin Secreting Adenoma; Pituitary Gland; Pituitary Neoplasms; Pituitary-dependent Cushing' s disease; Plasmids; Primary Neoplasm; Pro-Opiomelanocortin; Radiation therapy; Rare Diseases; Renilla Luciferases; Repeat Surgery; Reporter; Research Personnel; Residual Tumors; Series; Speed; Structure; System; Testing; Transactivation; Translating; Validation; Variant; Yeasts; base; cheminformatics; clinical investigation; cloud based; cost effectiveness; cross reactivity; cytotoxic; drug discovery; efficacious treatment; experience; high throughput screening; hypothalamic-pituitary-adrenal axis; in vivo; inhibitor/antagonist; internal control; multidisciplinary; novel; polypeptide; promoter; receptor; response; scaffold; small molecule; small molecule inhibitor; small molecule libraries; statistics; tool; transcription factor; tumor

ABSTRACT Cushing Disease (CD) is caused by an adrenocorticotropic hormone (ACTH)-secreting pituitary tumor that causes excess adrenal-derived cortisol. It is a life-threatening “orphan disease” with a staggering annual health care cost that is >7-fold higher than average patients. Surgical removal is the current first-line therapy but the disease frequently recurs. Repeat surgery, radiation therapy and bilateral adrenalectomy are not always successful and associated with major morbidity. Currently available drugs or those in clinical trials for CD do not target the pituitary corticotroph tumor itself and escape from control is common with long-term use. A clear unmet need for efficacious and safe therapies that offer biochemical and tumor control exists. We hypothesize that direct targeting of corticotroph tumors to modulate ACTH and in turn, glucocorticoid secretion is the optimal way to treat CD. We recently demonstrated that the orphan testicular receptor 4 (TR4, also known as NR2C2) is a potent regulator of hypothalamic-pituitary-adrenal (HPA) axis function and directly regulates pro- opiomelanocortin (POMC) gene transcription and ACTH secretion. We hypothesize that small molecule inhibitors of TR4 action would be potent inhibitors of corticotroph tumor hormone (ACTH) secretion. Such a discovery would be a transformative therapy for Cushing disease as no similar therapies exist. To identify and characterize TR4 small molecule modulators, we will perform a large scale small molecule library screen using our unique series of transactivation assays that include a TR4-directed GAL4-LBD system and POMC-promoter reporters as well as a secondary screen in human corticotroph tumor primary cultures. Our first aim will use a mammalian one-hybrid GAL4-LBD system to identify TR4 modulators from a library of 200,000 distinct compounds. Potential “hit” compounds will be confirmed by dose-response evaluation assays to calculate their EC50. Compounds identified in aim 1 will be further validated in aim 2 using a POMC transactivation assay using full-length and truncated TR4 variant plasmids together with a series of POMC promoter-driven luciferase reporters in murine corticotroph tumor AtT20 cells. A secondary screen will also be performed in human corticotroph tumor primary cultures to confirm their in vitro effects on ACTH secretion. We expect our proposal will identify and rigorously validate compounds that efficiently and specifically abrogate TR4 actions to inhibit ACTH secretion and lead to the discovery of safe and efficacious lead TR4 inhibitory compounds that can be further advanced as potential drug therapies for CD.

抽象的 库欣疾病（CD）是由肾上腺皮质激素（ACTH）引起的 - 定位垂体肿瘤 这会导致过量的肾上腺衍生的皮质醇。这是一种威胁生命的“孤儿疾病”，年度惊人 医疗费用比普通患者高7倍。手术去除是当前的一线疗法，但 该疾病经常复发。重复手术，放射治疗和双侧肾上腺切除术并不总是 成功并与主要发病率有关。目前可用的药物或CD临床试验中的药物没有 靶向垂体皮质营养肿瘤本身，而从控制中逃脱是常见的。清晰的未满足 需要有效且安全的疗法提供生化和肿瘤控制。我们假设这一点 直接靶向皮质营养肿瘤以调节ACTH，然后糖皮质激素分泌是最佳方式 处理CD。我们最近证明了孤儿检测受体4（TR4，也称为NR2C2）是一个 下丘脑 - 垂体 - 肾上腺（HPA）轴功能的有效调节剂，并直接调节促进剂 Opiomelananocortin（POMC）基因转录和ACTH分泌。我们假设小分子抑制剂 TR4作用的作用将是皮质营养肿瘤马（ACTH）分泌的有效抑制剂。这样的发现 由于没有类似的疗法，将是用于库欣疾病的变革疗法。识别和表征 TR4小分子调节剂，我们将使用我们的独特 一系列反式激活测定法，包括TR4定向的GAL4-LBD系统和POMC促销记者 以及人类皮质营养肿瘤原发性培养物中的次要筛查。我们的第一个目标将使用哺乳动物 一个杂交GAL4-LBD系统，可从200,000个不同化合物的库中识别TR4调节剂。潜在的 “命中”化合物将通过剂量反应评估测定法确认，以计算其EC50。化合物 在AIM 1中确定的AIM 2将使用POMC反式激活测定法进一步验证 截短的TR4变体质粒以及一系列POMC启动子驱动的荧光素酶记者在鼠中 皮质营养肿瘤ATT20细胞。还将在人皮质营养肿瘤原发性中进行次级屏幕 培养物以确认其对ACTH分泌的体外作用。我们希望我们的建议将确定和严格 验证有效，专门废除TR4动作以抑制ACTH分泌的化合物并导致 发现安全有效的铅TR4抑制化合物，可以进一步提高潜力 CD的药物疗法。