Stereochemical and dual peptide siRNA-carrier complexes for oral cancer therapy

用于口腔癌治疗的立体化学和双肽 siRNA 载体复合物

基本信息

批准号：
9375341
负责人：
Andrew George Jakymiw
金额：
$ 22.43万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-10 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9375341
关键词：
Accounting Address Amino Acids Anchorage-Independent Growth Animals Arginine Binding Biochemical Biological Availability Breast Cancer Etiology Cations Cells Cessation of life Chemicals Clinical Complex Data Development Disease Disease Outcome Dose Double-Stranded RNA Endosomes Gene Expression Genes Genetic Transcription Goals Head and Neck Cancer Human In Vitro Injection of therapeutic agent Intravenous Lead Legal patent Malignant Neoplasms Mammalian Cell Mediating Mediator of activation protein Methods Molecular Biology Neck Cancer Oncogenes Oral Stage Oral cavity Outcome Peptides Pharmaceutical Preparations Pharmacotherapy Pharyngeal structure Process Property Prostate Proteins Public Health RNA Degradation RNA Interference RNA Interference Therapy Regulator Genes Research Serum Skin Small Interfering RNA Small RNA Solid Neoplasm Specificity Survival Rate Technology Testing Therapeutic Tissues Toxic effect Translations Tumor Burden Tumor Tissue Untranslated RNA Work base cancer cell cancer therapy cancer type clinically relevant design improved improved outcome in vivo influenzavirus mRNA Transcript Degradation malignant mouth neoplasm mouse model mouth squamous cell carcinoma novel novel therapeutic intervention outcome forecast protein aminoacid sequence small molecule stereochemistry targeted delivery therapeutic effectiveness treatment strategy tumor growth

项目摘要

Although significant advances have been made in cancer treatment, the prognosis for oral cancer remains poor in comparison to other cancer types, including breast, skin, and prostate. Therefore, new therapeutic approaches are necessary to improve the outcome of this disease. The discovery that the introduction of chemically synthesized small interfering RNAs (siRNAs) into mammalian cells could efficiently induce sequence-specific inhibition of gene expression, made evident the therapeutic potential of harnessing RNA interference (RNAi) as a means to specifically target and silence disease-causing genes. Although the design of therapeutic-grade siRNAs has improved, delivery still remains the single greatest obstacle towards the pervasive use of siRNAs for therapeutic applications. Thus, to enhance the intracellular bioavailability of siRNAs in diseased tissues, effective new strategies for delivery are needed. Recently, we demonstrated that a novel peptide carrier we designed, termed 599, that comprised cell-penetrating and endosome-disruptive properties, could enhance the intracellular delivery and bioavailability of siRNAs designed to target the CIP2A oncogene (siCIP2A) into oral cancer cells in vitro, with intratumoral administration of the 599 peptide-siCIP2A complex inducing CIP2A silencing and consequently tumor growth inhibition in vivo. Despite these findings, further studies are still needed to better understand the importance in the chemical design of the 599 peptide and in determining the therapeutic effectiveness of administering the 599 peptide intravenously, since systemic delivery remains the standard method of administering drugs for the treatment of solid tumors. Consequently, because the 599 peptide was not cell/tissue-specific, we recently developed a dual peptide-mediated technology, where we found in our preliminary studies that combining a cancer cell-targeting peptide with the 599 peptide into a dual peptide-siCIP2A complex could mediate increased targeted delivery of siCIP2As into tumor tissues and significantly enhance CIP2A silencing upon systemic delivery. Therefore, based on the above findings, the goal of the current proposal is to further develop and improve upon the design of this 599 peptide-based siRNA delivery strategy for the treatment of oral cancer by determining the importance of amino acid chirality in 599 peptide design/function and whether systemic delivery of siCIP2As using the dual peptide- mediated technology can reduce tumor burden. To accomplish this goal, we will use biochemical and molecular biology approaches to: (1) determine the functional importance of stereochemistry in terms of 599 peptide-mediated intracellular delivery of bioactive siRNAs; and (2) determine the efficacy of the dual peptide- siCIP2A complex in mediating tumor growth inhibition in treated animals. The outcomes of the proposed research are expected to lead to improved chemical designs of the 599 peptide in mediating the delivery of bioactive siRNAs into cancer cells/tissues and in helping establish the therapeutic potential of the dual peptide- mediated technology for RNAi-based therapy in human oral cancer.

尽管癌症治疗已取得重大进展，但口腔癌的预后仍然存在与其他癌症类型（包括乳腺癌、皮肤癌和前列腺癌）相比，效果较差。因此，新的治疗需要采取一些方法来改善这种疾病的结果。发现引入化学合成的小干扰RNA（siRNA）进入哺乳动物细胞可以有效诱导基因表达的序列特异性抑制，证明了利用 RNA 的治疗潜力干扰（RNAi）作为特异性靶向和沉默致病基因的手段。虽然设计治疗级 siRNA 的数量已有所改善，但递送仍然是实现这一目标的最大障碍 siRNA 广泛用于治疗应用。因此，为了提高细胞内的生物利用度患病组织中的 siRNA 需要有效的新递送策略。最近，我们证明了我们设计了一种新型肽载体，称为 599，包含细胞穿透性和内体破坏性特性，可以增强针对 CIP2A 的 siRNA 的细胞内递送和生物利用度通过瘤内注射 599 肽-siCIP2A，在体外将癌基因 (siCIP2A) 转化为口腔癌细胞复合物诱导 CIP2A 沉默，从而抑制体内肿瘤生长。尽管有这些发现，仍需要进一步研究以更好地理解 599 肽化学设计的重要性并确定静脉注射 599 肽的治疗效果，因为全身性递送仍然是治疗实体瘤的标准给药方法。最后，因为 599 肽不是细胞/组织特异性的，所以我们最近开发了一种双肽介导的技术，我们在初步研究中发现，将癌细胞靶向肽与将 599 肽插入双肽-siCIP2A 复合物中可以介导 siCIP2A 的靶向递送增加肿瘤组织并在全身递送后显着增强 CIP2A 沉默。因此，基于第599章被人欺负了通过确定氨基的重要性，基于肽的 siRNA 递送策略治疗口腔癌第 599 章肽设计/功能中的酸性手性，以及是否使用双肽-系统递送 siCIP2As 介导技术可以减轻肿瘤负荷。为了实现这一目标，我们将利用生化和分子生物学方法：（1）确定立体化学的功能重要性（第 599 章）肽介导的生物活性 siRNA 的细胞内递送； (2) 确定双肽的功效 siCIP2A 复合物介导治疗动物的肿瘤生长抑制。拟议的结果研究预计将改进 599 肽的化学设计，以介导生物活性 siRNA 进入癌细胞/组织并帮助建立双肽的治疗潜力人类口腔癌基于 RNAi 的治疗介导技术。