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.

多形性胶质母细胞瘤是恶性胶质瘤最具侵袭性和最常见的表现。 以对现有治疗方式的抵抗为特征，并表现出神经衰弱的过程 最终导致死亡，通常在确诊后14个月内。预后很差，几乎100%失败 GBM药物开发速度，胶质瘤领域面临的关键挑战是识别和表征 新的药物靶点，以克服GBM臭名昭著的治疗耐药性，并开发药物输送平台 瞄准无法用药的基因损伤。恢复P53活性是一种有吸引力的治疗策略 对于GBM的治疗，AS~65%的原发GBM患者表达功能缺陷的野生型P53。 非典型Bcl2家族蛋白Bcl2L12(Bcl2-like-12)的扩增和过表达对p53的影响 通过阻断P53的转录活性发挥作用。为了抑制Bcl2L12的功能，我们建议使用新的 基于RNAi的纳米偶联物，称为球形核酸(SNAs)，中和Bcl2L12的表达 已确诊的神经胶质瘤。我们发现靶向Bcl2L12的SNAs(siBcl2L12-SNAs)能够穿越细胞 包括血脑屏障在内的膜。我们确定siBcl2L12-SNA不需要使用 有毒的辅助试剂，在细胞内和血-脑/血-瘤的交界处有效积聚 全身性递送时脑内胶质瘤的屏障。它们在生理环境中表现出稳定性， 刺激肿瘤内Bcl2L12基因和蛋白表达下调和P53重新激活，减少肿瘤生长 GBM PDX模型中的负担。建立SNA平台作为适用的P53激活治疗方式 对于其他高度恶性和致命的实体癌症的治疗，我们选择皮肤黑色素瘤作为 第二种癌症类型用于评价siBcl2L12-SNAs。与GBM类似，皮肤黑色素瘤 以罕见的P53突变和Bcl2L12表达升高为特征，这与 黑色素瘤的耐药性和进展。在这里，我们将测试Bcl2L12通过一种 高活性SNA结合物增强对P53的肿瘤抑制，减少GBM和黑色素瘤的进展，以及 这代表了一种新的、广泛适用的治疗策略，用于激活固体中的野生型p53 癌症。在目标1中，我们将通过以下方式确定P53重新激活的机制和确定替代标记 SiBcl2L12-SNAs。在目标2中，我们将优化SNA表面化学以优化siBcl2L12的传递 寡核苷酸对基底膜和黑色素瘤的作用。AIM 3将评估基因工程中的siBcl2L12-RNA 黑色素瘤(Aim 3a)和GBM小鼠模型(Aim 3b)，作为单一疗法，并与DNA联合使用 烷化剂替莫唑胺(目标3c)。这项提案的结果将提供对 Bcl2L12癌蛋白在细胞和生物学水平的表达，并将为成功实施多基因重组奠定基础 模式P53的重新激活作为治疗进入临床实践。

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