Effective analysis and manipulation of the protein corona - increasing the efficacy of Sixfold’s delivery system for Cell and Gene therapeutics

该项目将Six Double的可编程寡核苷酸传递系统(PODS)与LGC在蛋白质定量生物测量方面的独特和深厚的专业知识以及NPL在分析和测量纳米材料的物理化学和生物功能特性(包括先进成像)方面的专业知识结合在一起。豆荚已经证明了一种非常有希望的安全性，用于传递细胞和基因治疗药物，如用于基因沉默的短干扰RNA(SiRNA)。现在的目标是通过整合蛋白质日冕(PC)操作模块来优化siRNA-Pods的功效。为此，该项目采用了迭代的技术方法，利用了该联盟的跨学科专业知识，并利用了LGC/NPL的独特设施、设备和专门知识。这允许优化可无缝集成到高级治疗药物产品(ATMP)开发中的豆荚，使豆荚能够通过临床前开发快速进展，并加快商业化。鉴于其对基因沉默的高度特异性/选择性，siRNAs有可能为包括癌症在内的各种基因驱动的疾病提供有效的治疗选择。Alnylam的siRNA疗法于2018年首次获得监管部门的批准，这证实了此类疗法的临床和商业机会。它们进一步临床和商业成功的主要限制因素是缺乏系统地将siRNA输送到特定疾病细胞的安全和有效的系统。这一点得到了学术界和工业界的认可。目前的方法(主要是基于病毒和脂类的)由于其有限的特异性、毒性和复杂/昂贵的制造，限制了可治疗疾病的类型和数量，因此是次优的。PODS可以解决药物传递的挑战，因为它们基于中心纳米支架的独特的、生物兼容的设计，可以通过多种治疗方法和识别癌症细胞(但不是健康细胞)上的生物标志物的高度特异的靶向分子来实现功能。临床前数据表明，siRNA-Pods癌症治疗具有高度竞争的安全性。然而，基因沉默的效果仍有待优化。SiRNA-Pods的疗效受到静脉给药后PC在Pod上的重新募集的限制。PC可以显著影响纳米粒的物理性质、药代动力学和药效学[3]。通过利用LGC/NPL的专业知识，该项目实现了必要的高级(I)蛋白质组学、(Ii)形态和(Iii)PC结合豆荚的功能图谱，允许纳入优化的PC操作模块(S)，以提高治疗效果。这将加快临床前开发的完成，将从许可协议中产生第一笔收入的能力释放六倍，并加快临床开发，不仅是我们的主要适应症，也是其他疾病的临床开发，有助于提高英国自动药物治疗计划部门的竞争力。[1]Alnylam\_Press Release\_30.08.18.\[2\]Karim\_ME\_et\_al.\_Pharmaceutic\_2018.\[3\]Nguyen\_VH\_&\_Lee\_BJ\_Int\_J\_Nanomedicine\_2017\.

The project combines Sixfold's Programmable Oligonucleotide Delivery System (PODS) with the unique and deep expertise of LGC in quantitative bio-measurement of proteins, and NPL in analysis and measurement of physicochemical and biofunctional properties of nanomaterials, including advanced imaging. PODS have demonstrated a highly promising safety profile for delivery of Cell&Gene therapeutics such as short-interfering RNA (siRNA) for gene-silencing. The aim now is to optimise siRNA-PODS's efficacy by incorporating a protein corona (PC) manipulation module. To that end, the project employs an iterative technical approach, takes advantage of the interdisciplinary expertise of the consortium and leverages LGC's/NPL's unique facilities, equipment and know-how. This allows for optimisation of PODS that can be seamlessly integrated into the development of Advanced Therapy Medicinal Products (ATMPs), enabling PODS to quickly progress through preclinical development, and expediting commercialisation.Given their high specificity/selectivity for gene-silencing, siRNAs have the potential to provide effective treatment options for a variety of genetically-driven diseases including cancer. The first ever regulatory approval of Alnylam's siRNA therapy in 2018 \[1\] has validated the clinical and commercial opportunity for such therapies. The major limiting factor for their further clinical and commercial success is the lack of safe and effective systems for systemic delivery of siRNAs to specific diseased cells. This is recognised by academia and industry \[2\]. Current approaches (primarily viral- and lipid-based) are sub-optimal given their limited specificity, toxicity, and complex/expensive manufacturing \[2\], limiting the type and number of addressable disease indications.PODS can address the drug delivery challenge given their unique, biocompatible design based on a central nanoscaffold, which can be functionalised with multiple therapeutics and highly-specific targeting molecules that recognise biomarkers on cancer-but not healthy-cells.Preclinical data indicates a highly competitive safety profile of siRNA-PODS cancer therapy. However, the gene-silencing efficacy remains to be optimised. siRNA-PODS' efficacy is limited by recruitment of PC onto PODS upon intravenous administration. The PC can dramatically influence the physical properties, pharmacokinetics and pharmacodynamics of nanoparticles \[3\].By utilising LGC's/NPL's expertise, the project enables the necessary advanced (i) proteomic, (ii) morphological and (iii) functional profiling of PC-bound-PODS, allowing for incorporation of optimised PC manipulation module(s) for enhanced therapeutic efficacy. This would accelerate the completion of preclinical development, unlocking Sixfold's ability to generate first revenue from licensing agreements and expediting clinical development not only for our primary indication but also for other diseases, contributing to the competitiveness of the UK's ATMP sector.\[1\]Alnylam\_Press Release\_30.08.18.\[2\]Karim\_ME\_et\_al.\_Pharmaceutic\_2018.\[3\]Nguyen\_VH\_&\_Lee\_BJ\_Int\_J\_Nanomedicine\_2017\.