Peptide-mediated delivery of siRNA for treatment of ovarian cancer

肽介导的 siRNA 递送用于治疗卵巢癌

• 批准号: 8526659
• 负责人: Ester J Kwon
• 金额: $ 5.39万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8526659
• 关键词: Animal Model; Animals; Apoptosis; Apoptotic; Ascites; Binding; Biodistribution; Blood; Cancer Etiology; Cancer cell line; Cell Line; Cell surface; Cells; Cessation of life; Complex; Copy Number Polymorphism; Cytosol; Dana-Farber Cancer Institute; Detection; Development; Diagnosis; Disease; Disease model; Down-Regulation; Drug Formulations; Etiology; Flow Cytometry; Gene Mutation; Generations; Genetically Engineered Mouse; Heterogeneity; Human; Image; Immune; In Vitro; Individual; Innovative Therapy; Label; Luciferases; Malignant Neoplasms; Malignant neoplasm of ovary; Mammalian Oviducts; Measures; Mediating; Messenger RNA; Metabolic; Methods; Microscopy; Modeling; Monitor; Mus; Operative Surgical Procedures; Ovarian; Patients; Peptides; Prevention; Protein Isoforms; Proteins; Pyruvate Kinase; RNA Interference; Recurrence; Renal clearance function; Saline; Serum; Small Interfering RNA; Staging; Staining method; Stains; Surface; Survival Rate; TNF gene; Technology; Testing; Therapeutic; Translating; Tumor Burden; Western Blotting; Woman; Xenograft Model; Xenograft procedure; base; cancer cell; cancer genome; cancer therapy; caspase-3; chemotherapy; clinical application; clinically relevant; in vivo; inhibitor/antagonist; innovation; insight; interest; mouse model; nanoparticle; nanoparticulate; new therapeutic target; novel therapeutics; nuclease; particle; public health relevance; receptor; response; small molecule; subcutaneous; trafficking; tumor; tumor growth; uptake

DESCRIPTION (provided by applicant): Ovarian cancer is the fifth leading cause of cancer-related deaths in women. The majority of patients is diagnosed at late stages of disease due to inadequate detection methods, and is treated with surgery followed by chemotherapy. Due to the advanced stage of disease at diagnosis, there is a high rate of recurrence. The five-year survival rate of ovarian cancer is 20% below the average five-year survival rate for all cancer combined, highlighting the need for innovative therapies for ovarian cancer. Rapid and inexpensive sequencing technology has allowed for the ongoing efforts to profile individual cancer genomes and subsequent identification of new therapeutic candidates. However, the development of small molecule inhibitors against newly identified targets is expensive, slow, and many targets are "undruggable". RNAi therapy offers an attractive course of treatment: siRNA against new targets can be rapidly synthesized and can target specific isoforms which may not be possible by small molecule therapy. However, at this stage, efficient delivery of siRNA into the cytosol of diseased cells remains a major challenge. We propose utilizing primary ovarian cancer cells as a clinically relevant model of disease to optimize siRNA delivery vehicles for ovarian cancer therapy. There are several mouse models utilized to develop therapeutics for ovarian cancer, including genetically engineered mouse models and xenografts of ex vivo propagated cell lines. Although useful, these models may not represent the human etiology of high-grade ovarian cancer, the greater majority of which originate from the fallopian tube. Primary ovarian cells isolated from the ascites of human patients may more accurately represent disease development and the heterogeneity of cancer cells, and therefore may be more useful in evaluating the response to therapy. Characterization of primary ovarian cancers may provide insights to develop therapeutics that will be ultimately translated into humans. The purpose of this proposal is the development of siRNA delivery carriers to clinically relevant models of ovarian cancer. We will fine-tune our first generation carrier which includes (1) a peptide for targeting ectopically expressed p32 and (2) a peptide for mediating endosomal escape of internalized siRNA. First, we will characterize primary ovarian cancer cells for surface representation of the cognate receptor of our targeting peptide. Next, targeting peptide will be optimized for internalization and delivery of siRNA in primary ovarian cancer cells in vitro, as well as optimal biodistribution in vivo. These optimized vehicles will then be applied for therapeutic delivery of siRNA in primary ovarian cancer xenograft models. A multi-component peptide siRNA delivery vehicle has broad applicability to other cancer subtypes via interchanging targeting moieties and cargo.

描述(申请人提供)：卵巢癌是女性癌症相关死亡的第五大原因。由于检测方法不充分，大多数患者在疾病晚期被诊断出来，并在化疗后进行手术治疗。由于确诊时疾病处于晚期，复发率很高。卵巢癌的五年存活率比所有癌症的平均五年存活率总和低20%，这突显了卵巢癌创新疗法的必要性。快速和廉价的测序技术使正在进行的描绘单个癌症基因组和随后确定新的候选治疗方案的努力成为可能。然而，针对新确定的靶点的小分子抑制剂的开发是昂贵的、缓慢的，而且许多靶点是“无法下药的”。RNAi疗法提供了一个有吸引力的治疗过程：针对新靶点的siRNA可以快速合成，并可以靶向特定的异构体，这可能是小分子疗法无法实现的。然而，在这个阶段，有效地将siRNA运送到疾病细胞的胞浆中仍然是一个主要的挑战。我们建议利用原代卵巢癌细胞作为临床相关的疾病模型来优化用于卵巢癌治疗的siRNA载体。有几种小鼠模型被用来开发卵巢癌的治疗方法，包括基因工程小鼠模型和体外繁殖细胞系的异种移植。虽然这些模型有用，但它们可能并不能代表人类高级别卵巢癌的病因，因为大多数高级别卵巢癌起源于输卵管。从患者腹水中分离的原代卵巢细胞可能更准确地反映疾病的发展和癌细胞的异质性，因此在评估治疗反应方面可能更有用。原发卵巢癌的特征可能为开发最终将被移植到人类身上的治疗方法提供见解。这项建议的目的是开发siRNA递送载体以用于临床相关的卵巢癌模型。我们将微调我们的第一代载体，它包括(1)靶向异位表达的p32的多肽和(2)介导内化siRNA内体逃逸的多肽。首先，我们将表征原代卵巢癌细胞的表面表达我们的靶向多肽的同源受体。接下来，靶向多肽将在体外优化siRNA在原代卵巢癌细胞中的内化和递送，以及在体内的最佳生物分布。然后，这些优化的载体将被应用于在原发卵巢癌异种移植模型中治疗性地输送siRNA。多组份多肽siRNA载体通过互换靶向部分和货物，对其他癌症亚型具有广泛的适用性。