Accelerating early-phase development of novel therapeutic interventions: 'intelligent' anti-cancer agents against oncogenic microRNAs.

项目大纲。这项研究旨在开发新的治疗方法，通过受控的分子自组装和独特RNA序列的精确序列识别，靶向并不可逆转地破坏致病的、与癌症相关的microRNA分子。这些高度选择性的生物偶联物的催化性质将使我们能够提高未来治疗的效率，并将不良的偏离目标的影响降至最低。意义和潜在影响。一个主要的生物医学挑战是针对疾病状态(如癌症、炎症)中异常基因表达的高度选择性治疗，在这些疾病状态下，联合治疗，包括相对有毒的药物鸡尾酒，是另一种适应症。通过靶向上游细胞成分(如信使RNA、小的非蛋白质编码RNA、长的非编码RNA)实现对致病蛋白的可控翻译抑制，从而触发所需的治疗反应，可以促进选择性治疗疾病状态的新的治疗策略。事实上，短的功能非编码的microRNAs与许多类型的癌症有关，因此可以用作开发更具选择性和更强大的抗癌治疗的生物靶点。因此，RNA介导的基因沉默被认为是传统方法的一种有前途的替代方法，这些方法传统上基于在表达蛋白质水平上治疗生理异常，并且经常受到药物不良反应和毒性的影响。该项目的目标。该项目的重点是开发合成肽-寡核苷酸杂交物，用于选择性靶向具有扭曲表达谱的致病RNA序列(例如与癌症相关的microRNAs)。这些化学工程的RNA靶向分子将通过将催化不活跃的短肽与DNA识别基序结合而产生，以产生能够识别和切割与疾病相关的RNA的新型生物活性分子。这些分子最显著的特征是，多肽和寡核苷酸构建基团的结合协同结合了这两个组分的各自性质，产生了一种新的、具有特殊催化作用的杂化分子，能够在生理条件下切割RNA分子。这项研究的关键挑战将是开发新一代连接物，将RNA切割的精确序列特异性与增强的催化周转率相结合，以实现高效并减少脱靶效应。这一协调的跨学科项目将在化学生物学、生物物理学、结构生物学和药物输送的界面上进行。这种新型疗法的设计将基于详细的3D结构数据(以前通过核磁共振光谱和分子建模获得)和用于定点连接的新化学策略的协同组合。为了在这一早期开发阶段证明原则，我们将与Marina Zenkova教授(俄罗斯新西伯利亚化学生物学和基础医学研究所)和Michela Garofallo博士(英国曼彻斯特癌症研究所)合作，评估我们的构建物针对已建立的癌症相关microRNA序列小组的杂交和切割能力。该项目的主要成果是实现高水平的反应催化周转(‘分裂和离开’)，同时保持有效的生物专一性。多肽结构也将发生变化，以提供未来选择性靶向疾病组织的平台，并促进跨越生物屏障的运输。这项研究的成功将提供更好的治疗干预措施，提高效力，降低剂量，减少毒性。我们预计，这项研究的关键成果将为提高未来抗癌治疗的有效性和降低患者的风险提供基础。

Project Outline. This research aims to develop novel therapeutics to target and irreversibly destroy pathogenic, cancer-relevant microRNA molecules via controlled molecular self-assembly and precise sequence recognition of unique RNA sequences. The catalytic nature of these highly-selective bioconjugates will allow us to enhance the efficiency of future therapeutic treatments and minimise undesirable off-target effects. Significance and potential impact. A major biomedical challenge is highly-selective therapy against abnormal gene expression in disease states (e.g. cancer, inflammation) where combination therapies, including comparatively toxic drug cocktails, are otherwise indicated. Novel therapeutic strategies for selective treatment of disease states can be facilitated by targeting of upstream cellular components (e.g. messenger RNA, small non-protein-coding RNAs, long non-coding RNAs) to achieve controlled translational arrest of pathogenic proteins and thus trigger a desired therapeutic response. Indeed, short functional non-coding microRNAs are implicated in many types of cancer, and thus can be used as biological targets for development of more selective and powerful anticancer therapies. RNA-mediated gene silencing is recognised therefore as a promising alternative to the conventional approaches that are traditionally based on treatment of physiological abnormalities at the level of expressed proteins and which often suffer from adverse drug reaction and toxicity. Aims of the project. This project focuses on the development of synthetic peptidyl-oligonucleotide hybrids for selective targeting of pathogenic RNA sequences (e.g. cancer-related microRNAs) with distorted expression profiles. These chemically-engineered RNA-targeting molecules will be generated by conjugation of short, catalytically inactive peptides with DNA recognition motifs to produce novel biologically-active molecules capable of recognising and cleaving disease-relevant RNAs. The most remarkable feature of these molecules is that conjugation of peptide and oligonucleotide building blocks synergistically combines the individual properties of the two components, and yields a new, hybrid molecule with unusual catalysis, capable of cleaving RNA molecules under physiological conditions. The key challenge of this research will be to develop a new generation of conjugates that combine precise sequence-specificity of RNA cleavage with enhanced catalytic turnover to achieve high efficiency and reduce off-target effects.Proposed research plan. This coordinated cross-disciplinary project will be carried out at the interface of chemical biology, biophysics, structural biology and drug delivery. The design of this type of novel therapeutics will be based on a synergetic combination of the detailed 3D structural data (previously obtained from NMR spectroscopy and molecular modelling) and novel chemical strategies for site-directed conjugation. To demonstrate a proof-of-principle at this early-phase development, we shall evaluate hybridisation and cleavage capabilities of our constructs against established panel of cancer-relevant microRNA sequences in collaboration with Prof. Marina Zenkova (Institute of Chemical Biology & Fundamental Medicine, Novosibirsk, Russia) and Dr. Michela Garofallo (Manchester Cancer Research Institute, UK). The main deliverable output of this project is to achieve a high level of reaction catalytic turnover ('cleave and leave') while retaining effective bio-specificity. The peptide structure will also be varied to provide a future platform for selective targeting diseased tissue and facilitate transport across biological barriers. The success of this research will offer superior therapeutic interventions with improved potency, lower dosage and reduced toxicity. We anticipate that the key outputs of this research will provide a basis for enhanced efficacy of future anticancer therapeutics and reduced risk to patients.