Topically-delivered Targeted Gene Suppression of Immune Activation in Psoriasis

描述（由申请人提供）：AuraSense Therapeutics（AST）与西北大学沿着正在应用最近开发的技术，该技术使用谷胱甘肽功能化纳米颗粒（称为球形核酸或SNA）来控制蛋白质表达。这些容易合成的纳米颗粒可以具有DNA或RNA壳，并且流体动力学直径小于50 nm。SNA构建体是调节基因表达的一种强有力的新方式。传统药物开发是一个越来越昂贵的命题。近年来，制药商一直在积极努力开发基因调控的潜力，以产生癌症的治疗线索。靶基因的鉴定进展迅速，但制药行业作为一个整体面临着重大挑战，涉及安全和有效地使用基于阿托伐他汀的基因调控技术。主要障碍包括核酸稳定性、免疫相关毒性和递送挑战。寡核苷酸在血流中循环时和进入细胞后会发生降解。保持它们的稳定性对于任何基因调控策略的治疗应用都是至关重要的。不幸的是，传统基因调控中使用的“裸”基因序列不仅不稳定，而且会引发免疫反应，导致半衰期极短，疗效差。对于治疗皮肤病，局部递送是理想的，不仅通过将干预集中在疾病部位，而且通过使全身毒性的风险最小化;表皮屏障通常阻止核酸进入皮肤。产品SNA构建体由高度定向的寡核苷酸的密集球形阵列组成，以纳米颗粒为模板。这些材料已经被证明是显著的，因为它们容易进入细胞，抵抗核酸酶降解，具有低毒性并导致最小的免疫刺激。至关重要的是，SNA还能够穿透角质层并击倒表皮中的靶标，使其成为治疗皮肤病的潜在非常强大的工具。利用最近关于IL-17和IL-22细胞因子在银屑病发展中的关键作用的前沿发现，AST和Northwestern将设计同时靶向这些细胞因子（IL 17 RA和IL 22 RA 1）的皮肤受体的SNA作为银屑病的局部治疗。将在体外和疾病的小鼠模型中评价这些SNA的基因敲除效力、毒性和治疗效果。长期目标经过一段时间的初步开发，AST的目标是通过与大型制药公司，如Abbvie，诺华或默克合作，将SNA技术商业化。目的1：产生有效抑制小鼠和人角质形成细胞中的基因靶标的IL 17 RA/IL 22 RA 1 SNA。AST将设计和测试含有靶向小鼠和人IL 17 RA和IL 22 RA 1（银屑病炎症途径中的关键受体）的SNA的SiRNA。SNA的功效将通过用靶向SNA相对于用非靶向siRNA双链体功能化的SNA处理的角质形成细胞中mRNA和蛋白质水平的降低来测量。目的2：评价IL 17 RA、IL 22 RA 1和IL 17 RA/IL 22 RA 1杂合SNA对银屑病的重建人表皮（RHE）模型的作用。使用在组织学、生物化学和遗传学上类似于银屑病的精氨酸诱导的人3-D模型，将进一步评估目标1中鉴定的最有效的个体和杂交人SNA。SNA疗效将根据靶向受体和细胞因子活化下游产物的mRNA和蛋白质表达的减少以及银屑病中观察到的异常分化的纠正进行评价。目的3：在小鼠模型中评估IL 17 RA/IL 22 RA 1 SNA抑制银屑病表型的能力。我们将使用咪喹莫特诱导的模型测试如目的1和2中鉴定的最有效的IL 17 RA和IL 22 RA 1 SNA预防和治疗银肩病的能力。与第二阶段工作的关系：如果这些研究显示出希望，我们将进一步测试SNA在另外两种小鼠模型中的疗效（通过IL-23诱导小鼠耳中的银屑病和具有人表皮移植的人源化小鼠系统），启动临床前毒性研究，理想情况下，为这些SNA申请ID。

DESCRIPTION (provided by applicant): AuraSense Therapeutics (AST), along with Northwestern University, is applying a recently developed technology using oligonucleotide-functionalized nanoparticles (called spherical nucleic acids or SNAs) to control protein expression. These easily synthesized nanoparticles can have either DNA or RNA shells, and are less than 50 nm in hydrodynamic diameter. SNA constructs are a powerful new way of regulating gene expression. Significance Conventional drug development is an increasingly expensive proposition. In recent years, pharmaceutical manufacturers have pursued gene regulation's potential with aggressive efforts to generate therapeutic leads for cancer. Identification of target genes has proceeded quickly, but the pharmaceutical industry as a whole faces major challenges involving the safe and effective use of oligonucleotide-based gene regulation technology. The major barriers include nucleic acid stability, immune-related toxicity, and delivery challenges. Oligonucleotides are subject to degradation while circulating in the bloodstream and upon entry into cells. Preserving their stability is critical for therapeutic application of any gene regulation strategy. Unfortunately, the "naked" genetic sequences used in traditional gene regulation are not only unstable, but elicit an immune response, leading to extremely short half-lives and poor efficacy. For treating skin disorders, topical delivery is idea, not only by concentrating the intervention at the disease site, but also by minimizing the risk of systemic toxicity; the epidermal barrier has generally precluded the entry of nucleic acids into skin. The Product SNA constructs consist of a dense spherical array of highly oriented oligonucleotides, templated on a nanoparticle. These materials have already proven to be remarkable in that they readily enter cells, resist nuclease degradation, have low toxicity and lead to minimal immunostimulation. Critically, SNAs are also capable of penetrating the stratum corneum and knocking down targets in the epidermis, making them potentially very powerful tools to treat skin diseases. Leveraging recent cutting-edge discoveries about the key roles of IL-17 and IL-22 cytokines in the development of psoriasis, AST and Northwestern will design SNAs that concurrently target the skin-based receptors for these cytokines (IL17RA and IL22RA1) as topical treatment of psoriasis. These SNAs will be evaluated for gene knockdown potency, toxicity and therapeutic effect in vitro and in a mouse model of the disease. Long Term Goal After a period of initial development, AST aims to commercialize the SNA technology by partnering with a large pharmaceutical company such as Abbvie, Novartis or Merck. The Specific Aims of this proposal are the following: Aim 1: Generate IL17RA/IL22RA1 SNAs that efficiently suppress gene targets in mouse and human keratinocytes. AST will design and test SiRNA containing SNAs that target mouse and human IL17RA and IL22RA1, key receptors in the psoriasis inflammation pathway. The efficacy of the SNAs will be measured by the reduction in mRNA and protein levels in keratinocytes treated with targeted SNAs vs. SNAs functionalized with non-targeting siRNA duplexes. Aim 2: Evaluate the effect of IL17RA, IL22RA1 and IL17RA/IL22RA1 hybrid SNAs on a reconstituted human epidermis (RHE) model of psoriasis. Using a cytokine-induced human 3-D model that histologically, biochemically, and genetically resembles psoriasis, the most potent individual and hybrid human SNAs identified in Aim 1 will be further assessed. SNA efficacy will be evaluated based on the reduction in mRNA and protein expression of targeted receptors and the downstream products of cytokine activation, as well as by correction of the abnormal differentiation seen in psoriasis. Aim 3: Evaluate the abilit of IL17RA/IL22RA1 SNAs to suppress the psoriatic phenotype in a mouse model. We will test the ability of the most efficacious IL17RA and IL22RA1 SNAs, as identified in Aims 1 and 2, to prevent and treat psoriasis using an imiquimod-induced model. Relationship to Phase II Work: Should these studies show promise, we will further test the efficacy of SNAs in two additional mouse models (induction of psoriasis in mouse ears by IL-23 and a humanized mouse system with human epidermal grafting), initiate pre-clinical toxicity studies and, ideally, apply for an ID for these SNAs.