Overcoming Resistance Mechanisms in Hedgehog and Myc-amplified Medulloblastoma

克服 Hedgehog 和 Myc 扩增的髓母细胞瘤的耐药机制

基本信息

批准号：
10527353
负责人：
Donald Coulter
金额：
$ 40.07万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-01 至 2025-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10527353
关键词：
3-Dimensional Apolipoprotein E Apoptosis Benzene Binding Biodistribution Blood - brain barrier anatomy Brain Brain Neoplasms CRISPR/Cas technology Cells Cerebellum Chemoresistance Childhood Brain Neoplasm Clinical Trials Combined Modality Therapy Cyclin D1 Cytoplasmic Granules Development Dose Drug Combinations Drug Delivery Systems Drug Kinetics Drug Transport Encapsulated Erinaceidae Evaluation Formulation Genes Genetic Transcription Genetic Variation Hepatotoxicity Human Immune Evasion Implant Individual Injections Laboratories Ligands Liver MB03 MYC gene MYCN gene Malignant Neoplasms Metabolism Modeling Mus Mutate Mutation Neurons Organ Pathway interactions Patients Peptides Pharmaceutical Preparations Phosphatidylinositols Phosphotransferases Primary Neoplasm Proliferating Resistance SHH gene Signal Pathway System Therapeutic Toxic effect Transgenic Mice Transgenic Organisms Tumor Burden Tumor Immunity Tumorigenicity analog antitumor effect blood-brain barrier crossing cancer stem cell cell growth cell killing chemotherapy copolymer drug development effective therapy genetic makeup imaging approach in vivo imaging system inhibitor innovation kinase inhibitor medulloblastoma medulloblastoma cell line meter mouse model mutant nanoparticle nanopolymer neoplastic cell nerve stem cell neurotoxic neurotoxicity novel novel therapeutics patient derived xenograft model peptidomimetics programmed cell death ligand 1 resistance mechanism side effect small molecule smoothened signaling pathway stem cell proliferation targeted treatment technology platform therapeutic target tumor tumorigenic uptake

项目摘要

PROJECT SUMMARY Medulloblastoma (MB) is a pediatric brain tumor arising from the cerebellum. MB treatment is challenging due to diverse genetic make-up, resistance to chemotherapy, inefficient drug transport across the blood brain barrier (BBB) and drug induced neurotoxicity. Hedgehog (Hh) and IGF/PI3K signaling pathways regulate cell growth, cancer stem cell (CSC) proliferation, and tumorigenicity in MB patients. Hh inhibitors are effective initially to treat SHH-MB, but their repeated use develops chemoresistance due to mutations in SMOothened (SMO) but can be overcome by modulating GLI, which is downstream of SMO using SF2523, which is a BRD4/PI3K dual inhibitor and inhibits MYCN expression. In our preliminary studies, we synthesize SMO inhibitor 2-chloro-N1-[4-chloro-3-(2-pyridinyl) phenyl]-N4, N4-bis (2-pyridinyl methyl)-1, 4-benzene- dicarboxamide (MDB5). MDB5 and SF2523 effectively inhibited the proliferation of ONS-76 and HD-MB03 cells in a dose dependent manner, with significantly higher cell killing when these drugs were used in combination. Treatment of HD-MB03 cells with the combination of these two drugs showed significantly higher decrease in colony formation and cyclin D1 expression but higher increase in Bax expression, compared to individual drugs. We synthesized mPEG-b-PCC-g-DC copolymer, with 5.1±0.21 and 6.5±0.1% loading for MDB5 and SF2523 when formulated into nanoparticles (NPs). There was sustained drug release from NPs, wherein 100% of MDB5 was released in 50 h, but only 60% of SF2523 was released in 80 h. Targeted NPs were prepared by mixing COG-133-PEG-b-PCC-g-DC and mPEG-b-PCC-g-DC at 10/90, 20/80 and 30/70 ratios, with the highest cellular uptake at 30/70 ratio. Systemic administration of COG-133-NPs loaded SF2523 into orthotopic SHH-MB tumor bearing NSG mice resulted in significantly higher drug concentration in the brain at 6 and 24h post administration compared to non-targeted NPs loaded with this drug while systemic injection of free drug showed negligible drug concentration in the brain. Moreover, systemic administration of COG-133-NPs loaded with MDB5 and SF2523 resulted in decreased tumor burden compared to non-targeted NPs of MDB5 and SF2523 as determined by IVIS imaging, with no hepatic toxicity. Our hypothesis is that BRD4/PI3K and Hh signaling pathways exert control over CSC proliferation in SHH-MB and hence represents a target for therapeutic exploitation with small molecules which can inhibit these signaling pathways. Our specific aims are to i) evaluate the resensitization effects of MDB5 and SF2523 on SHH and MYC driven MB and patient derived xenograft (PDX) cells; ii) formulate MDB5 and SF2523 into NPs with COG-133 conjugation and determine biodistribution and systemic/organ toxicity; and iii) determine effects of targeted NPs loaded with SF2523 and MDB5 in SHH and MYC driven orthotopic, patient derived xenograft (PDX) and transgenic SmoA1 MB mouse models. Long- term significance. Successful completion of this project will provide a platform technology for treating SHH- MB and other brain tumors using this innovative NP-based combination therapy of Hh and BRD4/PI3K inhibitors.

项目概要髓母细胞瘤 (MB) 是一种由小脑引起的儿童脑肿瘤，治疗具有挑战性。由于不同的基因组成、对化疗的抵抗力、药物在血脑中的转运效率低下屏障（BBB）和药物诱导的神经毒性（Hh）和IGF/PI3K信号通路调节细胞。 Hh 抑制剂可有效抑制 MB 患者的生长、癌症干细胞 (CSC) 增殖和致瘤性。最初用于治疗 SHH-MB，但由于 SMOothened 突变，重复使用会产生化疗耐药性（SMO），但可以通过调制 GLI 来克服，GLI 是使用 SF2523 的 SMO 下游，SF2523 是 BRD4/PI3K双重抑制剂并抑制MYCN表达在我们的初步研究中，我们合成了SMO。抑制剂 2-氯-N1-[4-氯-3-(2-吡啶基)苯基]-N4, N4-双(2-吡啶基甲基)-1, 4-苯- 二甲酰胺 (MDB5) 和 SF2523 有效抑制 ONS-76 和 HD-MB03 的增殖。以剂量依赖性方式对细胞进行杀伤，当这些药物用于联合使用这两种药物对 HD-MB03 细胞的治疗效果显着更高。与相比，集落形成和细胞周期蛋白 D1 表达减少，但 Bax 表达增加更多我们合成了 mPEG-b-PCC-g-DC 共聚物，负载量为 5.1±0.21 和 6.5±0.1%。 MDB5 和 SF2523 当配制成纳米颗粒 (NP) 时，药物会从 NP 中持续释放。 MDB5在50小时内释放了100%，但SF2523在80小时内仅释放了60%。通过以 10/90、20/80 和 30/70 的比例混合 COG-133-PEG-b-PCC-g-DC 和 mPEG-b-PCC-g-DC 来制备，将 SF2523 负载的 COG-133-NP 全身给药时细胞摄取率最高。原位SHH-MB荷瘤NSG小鼠导致大脑中药物浓度显着升高给药后 6 小时和 24 小时与全身注射该药物时负载该药物的非靶向 NP 相比此外，全身施用 COG-133-NP 后，游离药物在大脑中的药物浓度可以忽略不计。与 MDB5 和 SF2523 的非靶向 NP 相比，装载 MDB5 和 SF2523 导致肿瘤负荷减少通过 IVIS 成像确定 SF2523 没有肝毒性，我们的假设是 BRD4/PI3K 和 Hh。信号通路对 SHH-MB 中的 CSC 增殖进行控制，因此代表了治疗的靶点我们的具体目标是 i) 评估可抑制这些信号通路的小分子。 MDB5 和 SF2523 对 SHH 和 MYC 驱动的 MB 和患者来源的异种移植物的再敏作用 (PDX) 细胞；ii) 将 MDB5 和 SF2523 配制为具有 COG-133 缀合的 NP，并确定生物分布和全身/器官毒性；和 iii) 确定负载 SF2523 和 MDB5 的靶向 NP 在 SHH 中的作用和 MYC 驱动的原位、患者来源的异种移植 (PDX) 和转基因 SmoA1 MB 小鼠模型。该项目的成功完成将为治疗SHH-提供平台技术。使用这种基于 NP 的 Hh 和 BRD4/PI3K 抑制剂的创新联合疗法治疗 MB 和其他脑肿瘤。