Nanoparticles With Surface Antibody Against CD98 and Carrying CD98 Small Interfering RNA Reduce Colitis in Mice

Abstract

BACKGROUND & AIMS Nanoparticles have been explored as carriers of small interfering RNAs (siRNAs), and might developed to treat inflammatory bowel disease (IBD). Overexpression of CD98 on the surface of colonic epithelial cells and macrophages promotes development and progression of IBD. We developed an orally delivered hydrogel that releases nanoparticles with single-chain CD98 antibodies on their surface (scCD98-functionalized) and loaded with CD98 siRNA (siCD98). We tested the ability of the nanoparticles to reduce levels of CD98 in colons of mice with colitis. METHODS scCD98-functionalized siCD98-loaded nanoparticles were fabricated using a complex coacervation technique. We investigated the cellular uptake and lysosome escape profiles of the nanoparticles in Colon-26 cells and RAW 264.7 macrophages using fluorescence microscopy. Colitis was induced by transfer of CD4+CD45RBhigh T cells to Rag−/− mice or administration of dextran sodium sulfate to C57BL/6 mice. Mice were then given hydrogel (chitosan and alginate) containing scCD98-functionalized nanoparticles loaded with siCD98 or scrambled siRNA (control) via gavage. RESULTS The scCD98-functionalized nanoparticles were approximately 200 nm in size and had high affinity for CD98-overexpressing cells. The scCD98-functionalized siCD98-loaded nanoparticles significantly reduced levels of CD98 in Colon-26 cells and RAW 264.7 macrophages, along with production of inflammatory cytokines (TNFα, IL6, and IL12). In mice with colitis, administration of the scCD98-functionalized siCD98-loaded nanoparticles reduced colon expression of CD98. Importantly, the severity of colitis was also reduced, compared with controls (based on loss of body weight, myeloperoxidase activity, inflammatory cytokine production, and histologic analysis). Approximately 24.1% of colonic macrophages (CD11b+CD11c−F4/80+) in the mice had taken up fluorescently labeled siRNA-loaded nanoparticles within 12 hr of administration. CONCLUSIONS Nanoparticles containing surface CD98 antibody and loaded with siCD98 reduce expression of this protein by colonic epithelial cells and macrophages; oral administration decreases the severity of colitis in mice. This nanoparticle in hydrogel (chitosan/alginate) formulation might be developed to treat patients with IBD.

RAG1/2 Knockout Pigs with Severe Combined Immunodeficiency

“…RAG1/2 knockout mice are both immunodeficient and characterized by the absence of mature B and T cells (7,8). Such mice are commonly used for immunological, inflammation, oncology, and stem cell transplantation studies (9)(10)(11)(12). Immunodeficient mice have also been used for the functional reconstitution of the human hematopoietic and immune systems by transplanting human hematopoietic stem cells (13).…”

Use of Single-Chain Antibody Derivatives for Targeted Drug Delivery

“…By shortening the linker between two adjacent VL and VH domains, they fail to assemble to form a regular scFv; however, they can assemble with of hydrogel-encapsulated scFv-loaded nanoparticles reduced the colon expression of CD98 and diminished the severity of colitis (37). These results all indicate that scFv plays a pivotal role in the specificity of the siRNA delivery system and can serve as a suitable targeting ligand on the surface of drug carriers to target CD98-expressing cells.…”

“…As CD98 plays an important role in progression of IBD, it can serve as a potential target for therapy of IBD patients. Xiao and colleagues took advantage of this cell surface antigen to specifically target colonic epithelial cells and macrophages to reduce colitis (37). They applied an anti-CD98 scFv on the surface of anti-CD98 siRNA-loaded nanoparticles.…”

RNA interference-based nanosystems for inflammatory bowel disease therapy

“…Therapeutic RNA molecules can be entrapped in chitosan-based matrices. Xiao et al 41 formulated CD98 siRNA into chitosan polymeric nanoparticles and conjugated it with single-chain CD98 antibodies on its surface ( Figure 2B). The chitosan was obtained from further deacetylation, and the final degree of deacetylation was 95.6%.…”

“…Pathological section of hematoxylin/eosin (H&E)-stained colonic sections showed that the treatment group had considerably less inflammation. 41 Chitosan is not soluble in the physiological environment. 43 This property is primarily because the d-glucosamine residue in chitosan has a pK a value of 6.5.…”