Delivery of CD44 shRNA/nanoparticles within cancer cells: perturbation of hyaluronan/CD44v6 interactions and reduction in adenoma growth in Apc Min/+ MICE.
Our studies have shown that constitutive interactions between hyaluronan and CD44 on tumor cells induces various anti-apoptotic cell survival pathways through the formation of a multimeric signaling complex that contains activated receptor tyrosine kinases. Inhibition of the hyaluronan-CD44 interactions on tumor cells by hyaluronan-CD44 interaction antagonists suppresses these activities by disassembling the complex. Although the anti-tumor activity of hyaluronan-oligosaccharides, a hyaluronan-CD44 interaction antagonist, is effective in sensitizing tumor cells to chemotherapeutic agents and reducing tumor growth in xenografts, hyaluronan-oligosaccharide alone was not effective in reducing tumor progression in Apc Min/+ mice. We now show in vitro and in vivo that targeted inhibition of the expression of CD44v6 depletes the ability of the colon tumor cells to signal through hyaluronan-CD44v6 interactions. First, we cloned oligonucleotides coding CD44v6 shRNA into a conditionally silenced pSico vector. Second, using pSico-CD44v6 shRNA and a colon-specific Fabpl promoter-driven Cre recombinase expression vector packaged into transferrin-coated nanoparticles, we successfully delivered the CD44v6 shRNA within pre-neoplastic and neoplastic colon malignant cells. Third, using the Apc Min/+ mice model, we demonstrated that inhibition of the CD44v6 expression reduces the signaling through a hyaluronan/CD44v6-pErbB2-Cox-2 interaction pathway and reduced adenoma number and growth. Together, these data provide insight into the novel therapeutic strategies of short hairpin RNA/nanoparticle technology and its potential for silencing genes associated with colon tumor cells.
Misra
Suniti
S
Department of Cell Biology and Anatomy, Medical University of South Carolina, Charleston, South Carolina 29425, USA. misra@musc.edu
Hascall
Vincent C
VC
De Giovanni
Carla
C
Markwald
Roger R
RR
Ghatak
Shibnath
S
D000236:Adenoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D000818:Animals; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D051546:Cyclooxygenase 2 / Q000096:biosynthesis / Q000235:genetics; D050556:Fatty Acid-Binding Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D015316:Genetic Therapy; D006801:Humans; D018960:Hyaluronan Receptors / Q000096:biosynthesis* / Q000235:genetics; D006820:Hyaluronic Acid / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D053758:Nanoparticles; D009368:Neoplasm Transplantation; D012313:RNA / Q000494:pharmacology*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000096:biosynthesis / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics; D014168:Transferrin / Q000494:pharmacology*
D051546:Cyclooxygenase 2; D050556:Fatty Acid-Binding Proteins; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid; D012313:RNA; D018719:Erb-b2 Receptor Tyrosine Kinases; D014168:Transferrin; C497483:CD44 protein, human; C476399:Cd44 protein, mouse; C493320:Fabp1 protein, mouse; C077623:Ptgs2 protein, mouse; C496540:PTGS2 protein, human; C508053:ERBB2 protein, human; C524056:Erbb2 protein, mouse