Systemic RNA interference to reactivate p53 tumor suppression

系统性 RNA 干扰重新激活 p53 肿瘤抑制

• 批准号: 10091404
• 负责人: CHAD A. MIRKIN
• 金额: $ 35.29万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-01 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10091404
• 关键词: Ablation; Activation Analysis; Adjuvant; Alkylating Agents; Animals; Antineoplastic Agents; Apoptosis; BRAF gene; Biodistribution; Biological; Blood; Blood - brain barrier anatomy; Brain; Cells; Cellular Membrane; Cessation of life; ChIP-seq; Chemistry; Clinical; Complex; Cutaneous Melanoma; DNA; DNA Damage; Diagnosis; Drug Delivery Systems; Drug Kinetics; Drug resistance; Environment; Evaluation; Exhibits; Failure; Gene Silencing; Generations; Genes; Genetic; Genetic Engineering; Genetic Transcription; Glioblastoma; Glioma; Goals; Histopathology; Knowledge; Lesion; Libraries; Malignant - descriptor; Malignant Glioma; Malignant Neoplasms; Mediating; Melanoma Cell; Messenger RNA; Mitochondria; Modality; Modeling; Molecular; Monitor; Mutation; Nanoconjugate; Nanotechnology; Neurologic; Nuclear; Oligonucleotides; Oncoproteins; Outer Mitochondrial Membrane; PTEN gene; Patients; Physiological; Platelet-Derived Growth Factor; Predisposition; Prognosis; Protein Family; Proteins; RNA Interference; Reagent; Regimen; Reporter; Research; Resistance; Second Primary Cancers; Small Interfering RNA; Solid; Solid Neoplasm; Spherical Nucleic Acids; Surface; Surrogate Markers; TP53 gene; Testing; Therapeutic; Time; Tissues; Toxic effect; Tumor Burden; Tumor Suppression; Tumor Suppressor Genes; Western Blotting; base; blood-brain tumor barrier; cancer type; chemotherapy; clinical practice; combinatorial; cytochrome c; design; drug development; effective therapy; genetic signature; immunogenicity; in vitro activity; in vivo; inhibitor/antagonist; innovation; knock-down; melanoma; mouse model; multimodality; mutant; nanotechnology platform; new therapeutic target; novel; overexpression; patient derived xenograft model; response; restoration; siRNA delivery; side effect; temozolomide; treatment strategy; tumor; tumor progression; uptake

Glioblastoma multiforme (GBM), the most aggressive and prevalent manifestation of malignant glioma, are characterized by resistance to extant therapeutic modalities, and exhibit a neurologically debilitating course culminating in death often within 14 months after diagnosis. With this dismal prognosis and a near 100% failure rate of GBM drug development, the critical challenges facing the glioma field are to identify and characterize new drug targets to overcome the notorious therapy resistance of GBM, and to develop drug delivery platforms to target undruggable genetic lesions. Restoration of p53 activity represents an attractive therapeutic strategy for the treatment of GBM, as ~65% of primary GBM patients express functionally defective wildtype p53. Amplification and overexpression of the atypical Bcl2 family protein Bcl2L12 (Bcl2-Like-12) compromises p53 function by blocking the transcriptional activity of p53. To inhibit Bcl2L12 function, we propose to use novel RNAi-based nanoconjugates, termed Spherical Nucleic Acids (SNAs) to neutralize Bcl2L12 expression in established glioma. We have found that Bcl2L12-targeting SNAs (siBcl2L12-SNAs) are able to traverse cellular membranes including the blood-brain-barrier. We established that siBcl2L12-SNAs do not require the use of toxic auxiliary reagents and accumulate effectively in cells and upon crossing of the blood-brain/blood-tumor barrier in intracerebral gliomas upon systemic delivery. They exhibit stability in physiological environments, provoke robust intratumoral Bcl2L12 mRNA and protein knockdown and p53 reactivation, and reduce tumor burden in GBM PDX models. To establish the SNA platform as a p53 activating therapeutic modality applicable to the treatment of other highly malignant and lethal solid cancers, we elected cutaneous melanoma as a second cancer type for the evaluation of siBcl2L12-SNAs. Similar to GBM, cutaneous melanoma are characterized by infrequent p53 mutation and elevated Bcl2L12 expression, which correlates with the degree of melanoma drug resistance and progression. Here, we will test the hypothesis that Bcl2L12 ablation by a high activity SNA conjugate increases p53 tumor suppression, reduces GBM and melanoma progression, and thus represent a novel, broadly applicable therapeutic strategy for the activation of wild-type p53 in solid cancers. In Aim 1, we will determine the mechanism and identify surrogate markers of p53 reactivation by siBcl2L12-SNAs. In Aim 2, we will optimize SNA surface chemistry for optimized delivery of siBcl2L12 oligonucleotides to GBM and melanoma tumors. Aim 3 will evaluate siBcl2L12-SNAs in genetically engineered melanoma (Aim 3a) and GBM mouse models (Aim 3b), as monotherapies, and in combination with the DNA alkylator temozolomide (Aim 3c). The results of this proposal will provide an in-depth characterization of the Bcl2L12 oncoprotein at cellular and biological levels, and will pave the way to successfully implement multi- modal p53 reactivation as therapy into clinical practice.

胶质母细胞瘤多形（GBM）是恶性神经胶质瘤的最具侵略性和最普遍的表现 以对现有治疗方式的抗性为特征，并表现出神经学使人衰弱的过程 诊断后14个月内经常在死亡中达到高潮。这种惨淡的预后和接近100％的失败 GBM药物开发的速率，神经胶质瘤领域面临的关键挑战是识别和表征 克服GBM臭名昭著的疗法抗性并开发药物输送平台的新药物靶标 靶向不良遗传病变。 p53活性的恢复代表了一种有吸引力的治疗策略 对于GBM的治疗，由于约65％的初级GBM患者表达了功能有缺陷的WildType p53。 非典型Bcl2家族蛋白BCl2L12（Bcl2-like-12）的扩增和过表达损害了p53 通过阻止p53的转录活性来函数。为了抑制BCl2L12功能，我们建议使用新颖 基于RNAi的纳米缀合物，称为球形核酸（SNA），以中和Bcl2l12的表达 已建立的神经胶质瘤。我们发现BCl2L12靶向SNA（SIBCL2L12-SNA）能够遍历细胞 包括血脑屏障在内的膜。我们确定SIBCL2L12-SNA不需要使用 有毒辅助试剂并有效地积累在细胞和血液肿瘤的越过时 全身传递后脑神经胶质瘤中的障碍。它们在生理环境中表现出稳定性， 激发肿瘤内Bcl2l12 mRNA和蛋白质敲低和p53重新激活，并减少肿瘤 GBM PDX型号的负担。将SNA平台建立为P53激活治疗方式适用 为了治疗其他高度恶性和致命的固体癌，我们选择了皮肤黑色素瘤 用于评估SIBCL2L12-SNA的第二种癌症类型。与GBM相似，皮肤黑色素瘤是 以不经常的p53突变和升高的Bcl2l12表达为特征，该表达与程度相关 黑色素瘤耐药性和进展。在这里，我们将检验以下假设： 高活性SNA结合物增加了p53肿瘤的抑制作用，降低了GBM和黑色素瘤的进展，并且 因此，代表了一种新颖的，广泛适用的治疗策略，用于激活固体中野生型p53 癌症。在AIM 1中，我们将通过 SIBCL2L12-SNA。在AIM 2中，我们将优化SNA表面化学以优化SIBCL2L12的递送 寡核苷酸到GBM和黑色素瘤肿瘤。 AIM 3将评估基因工程中的SIBCL2L12-SNA 黑色素瘤（AIM 3A）和GBM小鼠模型（AIM 3B），作为单层，并与DNA结合 烷基替莫唑胺（AIM 3C）。该提案的结果将提供深入的表征 Bcl2L12在细胞和生物学水平上的癌蛋白，并将为成功实施多种 模态p53重新激活作为临床实践的治疗。

项目成果

Hairpin-like siRNA-Based Spherical Nucleic Acids.

• DOI: 10.1021/jacs.1c12750
• 发表时间: 2022-02-23
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Vasher MK;Yamankurt G;Mirkin CA
• 通讯作者: Mirkin CA

Multi-antigen spherical nucleic acid cancer vaccines.

• DOI: 10.1038/s41551-022-01000-2
• 发表时间: 2023-07
• 期刊: Nature biomedical engineering
• 影响因子: 28.1

Spherical Nucleic Acids: Adding a New Dimension to Nucleic Acids and Clinical Chemistry.

球形核酸：为核酸和临床化学增添新维度。

• DOI: 10.1373/clinchem.2016.268516
• 发表时间: 2018
• 期刊: Clinical chemistry
• 影响因子: 9.3
• 作者: Mirkin,ChadA;Petrosko,SarahHurst
• 通讯作者: Petrosko,SarahHurst

In Vivo Behavior of Ultrasmall Spherical Nucleic Acids.

• DOI: 10.1002/smll.202300097
• 发表时间: 2023-03
• 期刊: Small
• 影响因子: 13.3
• 作者: Cassandra E. Callmann;Matthew K Vasher;Anindita Das;Caroline D. Kusmierz;C. Mirkin

A first-in-human phase 0 clinical study of RNA interference-based spherical nucleic acids in patients with recurrent glioblastoma.

• DOI: 10.1126/scitranslmed.abb3945
• 发表时间: 2021-03-10
• 期刊: Science translational medicine
• 影响因子: 17.1
• 作者: Kumthekar P;Ko CH;Paunesku T;Dixit K;Sonabend AM;Bloch O;Tate M;Schwartz M;Zuckerman L;Lezon R;Lukas RV;Jovanovic B;McCortney K;Colman H;Chen S;Lai B;Antipova O;Deng J;Li L;Tommasini-Ghelfi S;Hurley LA;Unruh D;Sharma NV;Kandpal M;Kouri FM;Davuluri RV;Brat DJ;Muzzio M;Glass M;Vijayakumar V;Heidel J;Giles FJ;Adams AK;James CD;Woloschak GE;Horbinski C;Stegh AH
• 通讯作者: Stegh AH