Overcoming Resistance Mechanisms in Hedgehog and Myc-amplified Medulloblastoma
Overcoming Resistance Mechanisms in Hedgehog and Myc-amplified Medulloblastoma
批准号:
10308723
负责人:
Donald Coulter
金额:
$40.01万
依托单位国家:
美国
项目类别:
财政年份:
2020
资助国家:
美国
项目状态:
未结题
起止时间:
2020-12-01 至 2025-11-30
关键词:
3-DimensionalApolipoprotein EApoptosisBenzeneBindingBiodistributionBlood - brain barrier anatomyBrainBrain NeoplasmsCRISPR/Cas technologyCellsCerebellumChemoresistanceChildhood Brain NeoplasmClinical TrialsCombined Modality TherapyCyclin D1Cytoplasmic GranulesDevelopmentDoseDrug CombinationsDrug Delivery SystemsDrug KineticsDrug TransportEncapsulatedErinaceidaeEvaluationFormulationGenesGenetic TranscriptionGenetic VariationHepatotoxicityHumanImmune EvasionImplantIndividualInjectionsLaboratoriesLigandsLiverMB03MYC geneMYCN geneMalignant NeoplasmsMetabolismModelingMusMutateMutationNeuronsOrganPathway interactionsPatientsPeptidesPharmaceutical PreparationsPhosphatidylinositolsPhosphotransferasesPolymersPrimary NeoplasmResistanceSHH geneSignal PathwaySystemTechnologyTherapeuticToxic effectTransgenic MiceTransgenic OrganismsTumor BurdenTumor ImmunityTumorigenicityanalogantitumor effectbasecancer stem cellcell growthcell killingchemotherapycopolymerdrug developmenteffective therapygenetic makeupimaging approachin vivo imaging systeminhibitorinnovationkinase inhibitormedulloblastomamedulloblastoma cell linemouse modelmutantnanoparticleneoplastic cellnerve stem cellneurotoxicneurotoxicitynovelnovel therapeuticspatient derived xenograft modelpeptidomimeticsprogrammed cell death ligand 1resistance mechanismside effectsmall moleculesmoothened signaling pathwaystem cell proliferationtargeted treatmenttherapeutic targettumortumorigenicuptake
中文摘要
项目摘要
髓母细胞瘤是一种发生于小脑的小儿脑肿瘤。MB治疗具有挑战性
由于不同的基因组成,对化疗的抵抗,药物在血脑中的运输效率低下,
屏障(BBB)和药物诱导的神经毒性。Hedgehog(Hh)和IGF/PI 3 K信号通路调节细胞凋亡
MB患者的生长、癌症干细胞(CSC)增殖和致瘤性。Hh抑制剂有效
最初用于治疗SHH-MB,但由于SMOothened中的突变,它们的重复使用产生了耐药性。
(SMO)但是可以通过使用SF 2523调节GLI来克服,所述GLI是SMO的下游,SF 2523是
BRD 4/PI 3 K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
缓蚀剂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(MDB 5)。MDB 5和SF 2523能有效抑制ONS-76和HD-MB 03的增殖
细胞以剂量依赖性方式,当这些药物用于
组合.用这两种药物的组合处理HD-MB 03细胞显示出显著更高的细胞毒性。
与对照组相比,
个别药物。我们合成了mPEG-b-PCC-g-DC共聚物,其负载量为5.1±0.21和6.5±0.1%,
MDB 5和SF 2523配制成纳米颗粒(NP)时的组合物。从纳米粒中持续释放药物,
其中MDB 5在50 h内释放100%,而SF 2523在80 h内仅释放60%。目标NP为
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制备,
在30/70比率下细胞摄取最高。将负载SF 2523的COG-133-NP全身施用到
原位SHH-MB荷瘤NSG小鼠导致脑中的药物浓度显著更高,
在全身注射给药后6和24小时,与负载有该药物的非靶向NP相比,
游离药物在脑中的药物浓度可忽略不计。此外,COG-133-NP的全身给药
与MDB 5和SF 2523的非靶向NP相比,负载MDB 5和SF 2523的NP导致肿瘤负荷降低,
通过IVIS成像确定SF 2523,无肝毒性。我们的假设是BRD 4/PI 3 K和Hh
信号传导途径对SHH-MB中的CSC增殖发挥控制作用,因此代表了治疗的靶点。
利用可以抑制这些信号通路的小分子。我们的具体目标是:(i)评估
MDB 5和SF 2523对SHH和MYC驱动的MB和患者来源的异种移植物的再致敏作用
(PDX)ii)将MDB 5和SF 2523配制成具有COG-133缀合的NP,并测定生物分布
iii)确定负载有SF 2523和MDB 5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物(PDX)和转基因SmoA 1 MB小鼠模型。长-
术语意义该项目的成功完成将为治疗SHH提供平台技术-
MB和其他脑肿瘤使用Hh和BRD 4/PI 3 K抑制剂的这种创新的基于NP的组合疗法。
英文摘要
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.
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会议论文
Overcoming Resistance Mechanisms in Hedgehog and Myc-amplified Medulloblastoma
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批准号:10527353
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项目类别:
-
资助金额:$40.07万
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财政年份:2020
-
负责人:Donald Coulter
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依托单位:
海外基金