Nanoparticle-induced gene-silencing for the suppression of inflammation in mouse models (edema, scleroderma, sepsis)

一种多功能纳米颗粒系统，用于局部和/或全身抑制NF-[kappa] bp65介导的促炎细胞因子/趋化因子在急性和慢性炎症疾病中的转录。如果成功，它将广泛适用于炎症风险高的患者，例如外科手术期间的患者。这种治疗系统还可以预防性地应用于患者，特别是针对令人担忧的抗生素耐药性问题。此外，它也可能对那些与传统疗法不相容的患者有所帮助。NF-[kappa]B是一种细胞内蛋白，常规抗体疗法无法解决，因此该项目将开发siRNA的纳米颗粒载体。为此，将开发多壳sirna负载的磷酸钙纳米颗粒。它们将被适当的细胞靶向分子（即抗体和肽）功能化。经过近红外染料偶联后，它们也可以在体内检测到。作为替代系统，负载siRNA的磷酸钙纳米颗粒将被纳入聚乳酸纳米颗粒中，以获得更高程度的siRNA对核酸酶的保护。多功能纳米颗粒系统的效率将首先在确定的靶细胞中进行研究，如t细胞、b细胞、单核/巨噬细胞和内皮细胞，在受控的培养条件下，然后在活体动物体内进行研究。关于体内研究，纳米颗粒治疗急性局部（如水肿）、急性全身（败血症）和慢性（硬皮病）炎症的能力将被研究。在所有这些病例中，NF-[kappa]B在疾病过程中起重要作用。

A multifunctional nanoparticle-based system for local and/or systemic inhibition of NF-[kappa]B p65-mediated transcription of pro-inflammatory cytokines/chemokines in acute and chronic inflammatory diseases will be synthesized and examined. If it is successful, it will be widely applicable to patients who demonstrate a high risk of inflammation, e.g. during surgical operations. This therapeutic system can also be prophylactically applied in patients, especially with regard to the alarming problem of antibiotic resistance. Moreover, it could be also helpful in patients who are incompatible with conventional therapies. NF-[kappa]B is an intracellular protein that cannot be addressed by conventional antibody therapies, therefore nanoparticulate carriers for siRNA will be developed in this project. For this purpose, multi-shell siRNA-loaded calcium phosphate nanoparticles will be developed. They will be functionalized with appropriate cell-targeting molecules, i.e. antibodies and peptides. After near-infrared dye conjugation, they will also be detectable in vivo. As alternative system, siRNA-loaded calcium phosphate nanoparticles will be incorporated into polylactic acid nanoparticles in order to obtain a higher degree of siRNA protection from nucleases. The efficiency of the multifunctional nanoparticle-based system will be studied first in defined target cells, i.e. T-cells, B-cells, monocytes/macrophages, and endothelial cells, under controlled culture conditions, and second in vivo in living animals. Regarding the in vivo study, the ability of the nanoparticles to treat an acute localized (e.g. edema), an acute systemic (sepsis) and a chronic (scleroderma) inflammation will be studied. In all these cases, NF-[kappa]B plays an important role in the course of the disease.