Exploiting Mutant PPM1D-induced Metabolic Defects with Nanoparticle Encapsulated NAMPT Inhibitors

利用纳米颗粒封装的 NAMPT 抑制剂利用突变 PPM1D 诱导的代谢缺陷

• 批准号: 10507757
• 负责人: Matthew A. Murray
• 金额: $ 4.37万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10507757
• 关键词: Address; Anabolism; Anatomy; Animal Model; Astrocytes; Biocompatible Materials; Biodistribution; Biological Assay; Biological Models; Biomedical Engineering; Blood - brain barrier anatomy; Bone Marrow; Brain; Brain Neoplasms; Brain Stem; Cancer cell line; Cause of Death; Cell Line; Cell model; Cells; Clinical; Clinical Trials; Communication; Convection; Coupled; CpG Islands; Databases; Defect; Development; Diffuse intrinsic pontine glioma; Dose; Dose Limiting; Drug Delivery Systems; Encapsulated; Enzymes; Excision; Formulation; Future; Genes; Goals; Human; Hypermethylation; In Vitro; Intracranial Neoplasms; Laboratories; Location; Malignant Childhood Neoplasm; Malignant Neoplasms; Malignant neoplasm of central nervous system; Metabolic; Methods; Modeling; Molecular Profiling; Mutation; Nature; Oncogenic; Open Reading Frames; Operative Surgical Procedures; PPM1D gene; Patients; Pharmaceutical Preparations; Phenotype; Phosphoric Monoester Hydrolases; Polyethylene Glycols; Polymers; Pontine structure; Pre-Clinical Model; Publishing; Radiation therapy; Research; Retina; Science; Site; Survival Rate; Techniques; Testing; Therapeutic; Time; Toxic effect; Tumor Volume; Work; Xenograft procedure; brain parenchyma; brain tissue; cell killing; chemotherapy; copolymer; effective therapy; enzyme biosynthesis; in vivo; in vivo Model; inhibitor; innovation; mutant; nanoparticle; neoplastic cell; novel; novel therapeutic intervention; palliative; pharmacologic; poly(lactic acid); pre-clinical; promoter; small molecule; standard of care; stem; success; tumor; tumor microenvironment

PROJECT SUMMARY Diffuse Intrinsic Pontine Glioma (DIPG) is a leading cause of death from pediatric cancer, with an abysmal <1% five-year survival rate. The lethal nature of this cancer stems from the lack of effective treatment options, and while radiotherapy is the current standard of care, it can be considered is palliative at best. It is difficult to use surgery or chemotherapy as these tumors form within the blood brain barrier (BBB), in the pons region of the brainstem. Therefore, significant effort has been directed towards understanding the molecular profiles of DIPG, which can be targeted for selective tumor cell killing in these tumors. Recently, the Bindra laboratory published a novel discovery that truncating mutations in PPM1D, which are commonly found in DIPG, induce a global CpG Island hypermethylation phenotype (CIMP)-like state. This mutation leads to a diverse range of cellular phenotypes, including metabolic defects which we believe can be exploited for a therapeutic gain. To this end, we found that mutant PPM1D induces promoter hypermethylation in the NAD+ biosynthesis gene NAPRT, an important enzyme required to produce NAD+. Our group demonstrated that mutant PPM1D-induced NAPRT silencing confers exquisite sensitivity to a class of drugs which target NAMPT, another key NAD+ enzyme. This work was recently published in Nature Communications (Fons et al., 2019). Based on these novel findings, and the desperate need for new DIPG therapies, we propose to develop NAMPT inhibitors (NAMPTi’s) for the treatment of PPM1D-mutant DIPG. Blood-brain barrier (BBB)-penetrant NAMPTi’s have been developed and tested in clinical trials, however, their success has been limited by dose-limiting toxicities in the bone marrow and retina. To address these liabilities, this proposal aims to develop nanoparticle-encapsulated NAMPT inhibitors (NAMPTi-NPs) for direct delivery into the tumor microenvironment, and to validate them using in vitro and in vivo patient-derived models. My proposal will accomplish this goal with two aims: In Aim 1, we will create and optimize NPs that have high drug encapsulation efficiency, sustained retention of NAMPTi-NP in target sites, and demonstrate effective cell killing in NHA isogenic cell models and CCLE cell lines. Aim 2 will show the clinical feasibility of NAMPTi-NP via in vivo modeling that will encompass toxicity screens of free drug NAMPTi and NAMPTi-NP, as well as the biodistribution of these drugs within the brain. Lastly, we will show efficacy through reduction in size of PPM1D-mutant tumors treated with NAMPTi-NP. Altogether, this work will utilize a comprehensive and rigorous bench-based approach, coupled with computational and bioengineering methods, to develop a novel treatment for DIPG. My studies will serve as a critical proof-of- concept that seeks to establish an entirely new NP-based strategy targeting a key metabolic defect in DIPG. If successful, our approach can be applied to many other small molecules and brain tumors in the future.

项目摘要 弥漫性内在脑桥胶质瘤（DIPG）是儿童癌症死亡的主要原因， 五年生存率。这种癌症的致命性源于缺乏有效的治疗方案， 虽然放射治疗是目前的标准治疗，但它最多只能被认为是姑息治疗。很难用 手术或化疗，因为这些肿瘤在血脑屏障（BBB）内形成，在脑桥区域， 脑干因此，大量的努力已经指向理解DIPG的分子谱， 其可以被靶向用于选择性杀死这些肿瘤中的肿瘤细胞。最近，宾德拉实验室发表了 一项新的发现，即在DIPG中常见的PPM 1D中的截短突变诱导了全局CpG 岛状高甲基化表型（CIMP）样状态。这种突变导致多种细胞 表型，包括我们认为可以利用的代谢缺陷来获得治疗效果。为此目的， 我们发现突变型PPM 1D诱导NAD+生物合成基因NAPRT启动子的超甲基化， 产生NAD+所需的重要酶。我们的研究小组证明突变型PPM 1D诱导的NAPRT 沉默赋予了对靶向NAMPT（另一种关键NAD+酶）的一类药物的高度敏感性。这 工作最近发表在Nature Communications（Fons等人，2019年）。基于这些新的发现， 迫切需要新的DIPG疗法，我们建议开发NAMPT抑制剂（NAMPTi）， PPM 1D突变体DIPG的处理。已经开发了血脑屏障（BBB）渗透剂NAMPTi， 然而，在临床试验中，它们的成功受到骨髓剂量限制性毒性的限制。 和视网膜。为了解决这些责任，本提案旨在开发纳米颗粒封装的NAMPT 抑制剂（NAMPTi-NP）直接递送到肿瘤微环境中，并使用 体外和体内患者来源的模型。我的建议将通过两个目标实现这一目标：在目标1中，我们 将产生和优化具有高药物包封效率、NAMPTi-NP在药物包封中的持续保留的NP。 靶向位点，并在NHA同基因细胞模型和CCLE细胞系中证明有效的细胞杀伤。目标2将 通过体内建模显示NAMPTi-NP的临床可行性，其将包括游离药物的毒性筛选 NAMPTi和NAMPTi-NP，以及这些药物在脑内的生物分布。最后，我们将展示 通过减小用NAMPTi-NP处理的PPM 1D突变肿瘤的尺寸来提高疗效。总之，这项工作将 利用全面和严格的基于基准的方法，加上计算和 生物工程方法，开发一种新的治疗DIPG。我的研究将作为一个关键的证据- 该概念旨在建立一种全新的基于NP的策略，靶向DIPG中的关键代谢缺陷。如果 如果成功，我们的方法将来可以应用于许多其他小分子和脑肿瘤。