Targeting CDK6 expression/activity in Ph+ and Ph1-like acute lymphoblastic leukemia (ALL)

靶向 Ph 和 Ph1 样急性淋巴细胞白血病 (ALL) 中的 CDK6 表达/活性

• 批准号: 10652495
• 负责人: BRUNO CALABRETTA
• 金额: $ 61.87万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10652495
• 关键词: Acute; Acute Lymphocytic Leukemia; Adult; B-Cell Acute Lymphoblastic Leukemia; Biological; Blood; CDK4 gene; Cancer Biology; Catalytic Domain; Cell Cycle; Cell Cycle Proteins; Cell Line; Clinic; Clinical; Combination Drug Therapy; Complex; Cyclin D1; Cyclin-Dependent Kinase Inhibitor 2A; Cytoplasm; Development; Dose; Down-Regulation; Drug Kinetics; Exhibits; FDA approved; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Transcription; Goals; Growth; HDAC1 gene; Half-Life; Hematologic Neoplasms; Human; Imatinib; In Vitro; Intervention; Knowledge; Laboratories; Lead; Link; Liver Microsomes; MLL-rearranged leukemia; Malignant Neoplasms; Metabolic; Metabolic Pathway; Mitochondria; Molecular; Mus; Mutation; Nuclear; Oral; Outcome; Patients; Pharmaceutical Chemistry; Philadelphia; Phosphotransferases; Plasma; Prognosis; Proliferating; Property; Protac; Recurrent disease; Relapse; Resistance; Resistance development; Role; Route; S phase; Specificity; Structure-Activity Relationship; Subgroup; Testing; Therapeutic Agents; Therapeutic Effect; Tyrosine Kinase Inhibitor; acute lymphoblastic leukemia cell; advanced breast cancer; cancer therapy; cell growth; cyclin-dependent kinase 6; effective therapy; high risk; high throughput screening; improved; improved outcome; in vitro activity; in vivo; inhibitor; innovation; intraperitoneal; leukemia; leukemia treatment; mouse model; novel; novel therapeutics; patient derived xenograft model; pharmacologic; pre-clinical; prototype; therapeutically effective; validation studies

Abstract Philadelphia-positive acute lymhoblastic leukemia (Ph+ ALL) and Ph1-like B-ALL account for most cases of “high-risk” adult B-ALL. Current therapies with tyrosine kinase inhibitors (TKIs) have improved the outcome of Ph+ ALL, but resistance to TKIs develops rapidly in most patients. Ph1-like B-ALL is currently treated with intensive combination chemotherapy but disease relapse is common with a 5-year survival in only ~25% of patients. As a result, the prognosis of Ph+ ALL and Ph1-like B-ALL remains dismal. In previous studies, we showed that Ph+ and Ph1-like ALL cells exhibit a selective requirement for CDK6 expression while CDK4 expression is dispensable. CDK6 is the catalytic subunit of the cyclin D/CDK6 complex which is essential for the G1 to S-phase cell cycle transition and has kinase-independent growth-promoting effects in hematological malignancies. Our preliminary studies indicate that CDK6 silencing is more effective than CDK6 enzymatic inhibition in suppressing Ph+ ALL in mice. To block kinase-dependent and independent effects of CDK6, we have developed CDK4/6-targeted proteolysis-targeting chimera (PROTACs) that inhibit CDK4/6 enzymatic activity in vitro and promote the preferential degradation of CDK6 over CDK4 in Ph+ and Ph1-like ALL cells, providing durable suppression of CDK6 function. In this proposal, we will assess the requirement of CDK6 in Ph+ and Ph1-like ALL by comparing the effects of CDK6 degradation by PROTAC YX-2-107 and pharmacological inhibition using Palbociclib, an FDA-approved CDK4/6 inhibitor (Aim 1.1). We will also determine whether the more potent leukemia suppression induced by CDK6 down-regulation in comparison to CDK6 enzymatic inhibition can be explained by changes in gene expression induced selectively by CDK6 silencing. Such changes involve the histone deacetylase 1(HDAC1) gene and several others involved in mitochondrial metabolic pathways (Aims 1.2 and 1.3). Although we have been able to achieve high specificity of CDK6 versus CDK4 targeting and biological/therapeutic effects comparable/superior to Palbociclib ex vivo and in PDXs of Ph+ ALL, we will continue to improve our lead compound PROTAC YX-2-107 by medicinal chemistry approaches in order to develop derivatives with enhanced in vivo efficacy. In Aim 2, we will assess metabolic properties of select CDK6-degrading PROTACs and test their biological/therapeutic effects in Ph+ and Ph1-like ALL cells ex vivo and in mice injected with de novo or relapsed/TKI-resistant patient-derived Ph+/Ph1-like ALL cells. Collectively, our PROTAC-based approach which leverages the expertise in cancer biology and medicinal chemistry of the Calabretta and Salvino's laboratories holds promise to develop novel and more effective therapeutic agents for the treatment of CDK6-dependent high-risk B-ALL in pre-clinical PDX models and, potentially, in the clinic.

抽象的 费城阳性急性淋巴细胞白血病 (Ph+ ALL) 和 Ph1 样 B-ALL 占大多数病例 “高风险”成人 B-ALL。目前的酪氨酸激酶抑制剂（TKI）疗法已改善了以下疾病的治疗结果： Ph+ ALL，但大多数患者对 TKI 的耐药性迅速发展。 Ph1 样 B-ALL 目前的治疗方法是 强化联合化疗，但疾病复发很常见，只有约 25% 的患者能获得 5 年生存期 患者。因此，Ph+ ALL 和 Ph1 样 B-ALL 的预后仍然很差。在之前的研究中，我们 显示 Ph+ 和 Ph1 样 ALL 细胞对 CDK6 表达表现出选择性要求，而 CDK4 表达是可有可无的。 CDK6 是细胞周期蛋白 D/CDK6 复合物的催化亚基，对于 G1 期到 S 期细胞周期的转变，并在血液学中具有激酶独立的生长促进作用 恶性肿瘤。我们的初步研究表明 CDK6 沉默比 CDK6 酶促更有效 抑制小鼠 Ph+ ALL 的抑制作用。为了阻断 CDK6 的激酶依赖性和非依赖性作用，我们 开发了 CDK4/6 靶向蛋白水解嵌合体 (PROTAC)，可抑制 CDK4/6 酶 体外活性并促进 Ph+ 和 Ph1 样 ALL 细胞中 CDK6 相对于 CDK4 的优先降解， 提供对 CDK6 功能的持久抑制。在本提案中，我们将评估 CDK6 的要求 Ph+ 和 Ph1 样 ALL 通过比较 PROTAC YX-2-107 和 CDK6 降解的影响 使用 FDA 批准的 CDK4/6 抑制剂 Palbociclib 进行药理学抑制（目标 1.1）。我们还将 确定与 CDK6 下调相比，CDK6 下调是否能更有效地抑制白血病 CDK6 酶抑制可以通过 CDK6 选择性诱导的基因表达变化来解释 沉默。这种变化涉及组蛋白脱乙酰酶 1 (HDAC1) 基因和其他几个参与 线粒体代谢途径（目标 1.2 和 1.3）。尽管我们已经能够实现高特异性 CDK6 与 CDK4 靶向的比较以及生物/治疗效果与 Palbociclib 体外相当/优于 Palbociclib 在 Ph+ ALL 的 PDX 中，我们将继续通过药物改进我们的先导化合物 PROTAC YX-2-107 化学方法以开发具有增强体内功效的衍生物。在目标 2 中，我们将评估 选择 CDK6 降解 PROTAC 的代谢特性并在 Ph+ 中测试其生物/治疗效果 和 Ph1 样 ALL 细胞离体以及在从头注射或复发/TKI 耐药患者来源的小鼠体内 Ph+/Ph1 样 ALL 细胞。总的来说，我们基于 PROTAC 的方法利用了癌症方面的专业知识 卡拉布雷塔和萨尔维诺实验室的生物学和药物化学有望开发出新的药物 以及更有效的治疗药物用于治疗临床前PDX中CDK6依赖性高危B-ALL 模型，并有可能应用于临床。

项目成果

Inability to switch from ARID1A-BAF to ARID1B-BAF impairs exit from pluripotency and commitment towards neural crest formation in ARID1B-related neurodevelopmental disorders.

• DOI: 10.1038/s41467-021-26810-x
• 发表时间: 2021-11-09
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Pagliaroli L;Porazzi P;Curtis AT;Scopa C;Mikkers HMM;Freund C;Daxinger L;Deliard S;Welsh SA;Offley S;Ott CA;Calabretta B;Brugmann SA;Santen GWE;Trizzino M
• 通讯作者: Trizzino M