Silencing of proinflammatory genes targeted to peritoneal-residing macrophages using siRNA encapsulated in biodegradable microspheres.
One of the more substantial hurdles to be overcome in realizing the exciting potential of siRNA molecules as therapeutic agents for a wide range of diseases is the intact delivery of the active molecule into its target cell. Here, we present a platform for in vitro and in vivo delivery and intracellular release of siRNA in peritoneal macrophages (Mvarphis). The delivery platform is based on the encapsulation of siRNA in biodegradable poly(d,l-lactide) (PLA) microspheres, which are targeted to Mvarphis by the simple principle of size exclusion. Proof of concept was achieved using siRNAs targeting TNFalpha and CD86 in macrophages. We show that the release of the siRNA in peritoneal-derived macrophages in vitro occurs intracellularly, and is abrogated by cytochalasin B, a phagocytosis inhibitor. Silencing in these cells is potent and lasts for at least one week. In vivo, we prove that siRNA encapsulated in biodegradable PLA microspheres can be delivered to peritoneal-residing Mvarphis and can induce potent silencing of TNFalpha secretion for at least one week. The PLA microspheres hold great potential for in vivo use, due to their biocompatibility and degradability, and can potentially be used for in vivo immunomodulation of Mvarphis for treatment of autoimmune and chronic inflammatory conditions.
Brunner
Tali
T
Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer Sheva, Israel.
Cohen
Smadar
S
Monsonego
Alon
A
D000818:Animals; D051940:B7-2 Antigen / Q000378:metabolism; D001672:Biocompatible Materials / Q000378:metabolism*; D020868:Gene Silencing; D018836:Inflammation Mediators / Q000378:metabolism*; D042541:Intracellular Space / Q000378:metabolism; D019344:Lactic Acid / Q000378:metabolism; D017737:Macrophages, Peritoneal / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D009928:Organ Specificity; D011091:Polyesters; D011094:Polyethyleneimine / Q000378:metabolism; D011108:Polymers / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D051940:B7-2 Antigen; D001672:Biocompatible Materials; D018836:Inflammation Mediators; D011091:Polyesters; D011108:Polymers; D034741:RNA, Small Interfering; D014409:Tumor Necrosis Factor-alpha; D019344:Lactic Acid; C033616:poly(lactide); D011094:Polyethyleneimine