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

该项目结合了Sixfold的可编程寡核苷酸传递系统（PODS）， LGC在蛋白质定量生物测量方面的独特而深厚的专业知识，以及NPL在纳米材料的物理化学和生物功能特性分析和测量方面的专业知识，包括先进的成像。pod已被证明在细胞和基因疗法（如用于基因沉默的短干扰RNA (siRNA)）的递送方面具有很高的安全性。现在的目标是通过结合蛋白质冠（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，利用了联盟的跨学科专业知识，并利用了LGC /NPL独特的设施、设备和专业知识。这允许优化PODS，可以无缝集成到先进治疗药物产品（atmp）的开发中，使PODS能够通过临床前开发快速进展，并加快商业化。鉴于sirna对基因沉默的高特异性/选择性，sirna有可能为包括癌症在内的各种遗传驱动疾病提供有效的治疗选择。Alnylam的siRNA疗法于2018年首次获得监管机构批准\[1\]，这证实了此类疗法的临床和商业机会。限制其进一步临床和商业成功的主要因素是缺乏安全有效的sirna系统递送到特定病变细胞的系统。这得到了学术界和工业界的认可\[2\]。目前的方法（主要是基于病毒和脂质）由于其有限的特异性、毒性和复杂/昂贵的制造\[2\]，限制了可解决的疾病适应症的类型和数量。pod可以解决药物递送的挑战，因为它们独特的、基于中心纳米支架的生物相容性设计，可以用多种治疗方法和高度特异性的靶向分子进行功能化，这些分子可以识别癌症（但不是健康细胞）上的生物标志物。临床前数据表明，siRNA-PODS癌症治疗具有高度竞争性的安全性。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时PC在PODS上的招募的限制。PC可以显著影响纳米颗粒的物理性质、药代动力学和药效学\[3\]。通过利用LGC /NPL的专业知识，该项目能够对PC结合的pod进行必要的先进(i)蛋白质组学，（ii）形态学和（iii）功能分析，允许整合优化的PC操作模块，以提高治疗效果。这将加速临床前开发的完成，释放Sixfold从许可协议中获得首批收入的能力，加快临床开发，不仅针对我们的主要适应症，也针对其他疾病，有助于提高英国ATMP行业的竞争力。\[1\] Alnylam＿Press Release＿30.08.18。\[2\] Karim＿ME＿et＿al.＿Pharmaceutic＿2018。\[3\] 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\.