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

• 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
• 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

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.

虽然在癌症治疗方面已经取得了重大进展，但口腔癌的预后仍然存在 与其他癌症类型相比，包括乳腺癌，皮肤癌和前列腺癌。因此，新的治疗 必须采取措施来改善这种疾病的结果。我们发现， 化学合成的小干扰RNA（siRNA）进入哺乳动物细胞可以有效地诱导 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 尽管治疗级siRNA的数量有所改善，但递送仍然是实现治疗的最大障碍。 siRNA在治疗应用中的广泛应用。因此，为了提高细胞内的生物利用度， 在疾病组织中，需要有效的新递送策略。最近，我们证明， 我们设计了一种新的肽载体，称为599，它包括细胞穿透和内体破坏， 特性，可以增强设计用于靶向CIP 2A的siRNA的细胞内递送和生物利用度。 癌基因（siCIP 2A）导入口腔癌细胞，肿瘤内给予599肽-siCIP 2A 复合物诱导CIP 2A沉默并因此抑制体内肿瘤生长。尽管有这些发现， 仍然需要进一步的研究来更好地理解599肽在化学设计中的重要性。 以及确定静脉内施用599肽的治疗效果，因为全身性 递送仍然是用于治疗实体瘤的药物给药的标准方法。因此，委员会认为， 因为599肽不是细胞/组织特异性的，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肽引入到双肽-siCIP 2A复合物中可以介导增加的siCIP 2A向 并且在全身递送后显著增强CIP 2A沉默。因此，根据 根据上述调查结果，当前提案的目标是进一步开发和改进599 基于肽的siRNA递送策略通过确定氨基酸的重要性用于治疗口腔癌 599肽设计/功能中的酸手性以及是否使用双肽系统递送siCIP 2A- 介导的技术可以减少肿瘤负荷。为了实现这一目标，我们将使用生物化学和 分子生物学方法：（1）确定立体化学在599方面的功能重要性 肽介导的生物活性siRNA的细胞内递送;和（2）确定双肽-siRNA的功效。 siCIP 2A复合物介导治疗动物中的肿瘤生长抑制。拟议会议的成果 预期研究将导致599肽在介导药物递送中的改进的化学设计。 生物活性siRNA进入癌细胞/组织，并帮助建立双肽的治疗潜力- 介导的技术用于人类口腔癌的基于RNAi的治疗。