title: Targeted delivery of siRNA to cell death proteins in sepsis.
abstract: Immune suppression is a major cause of morbidity and mortality in the patients with sepsis. Apoptotic loss of immune effector cells such as CD4 T and B cells is a key component in the loss of immune competence in sepsis. Inhibition of lymphocyte apoptosis has led to improved survival in animal models of sepsis. Using quantitative real-time polymerase chain reaction of isolated splenic CD4 T and B cells, we determined that Bim and PUMA, two key cell death proteins, are markedly upregulated during sepsis. Lymphocytes have been notoriously difficult to transfect with small interfering RNA (siRNA). Consequently a novel, cyclodextrin polymer-based, transferrin receptor-targeted, delivery vehicle was used to coadminister siRNA to Bim and PUMA to mice immediately after cecal ligation and puncture. Antiapoptotic siRNA-based therapy markedly decreased lymphocyte apoptosis and prevented the loss of splenic CD4 T and B cells. Flow cytometry confirmed in vivo delivery of siRNA to CD4 T and B cells and also demonstrated decreases in intracellular Bim and PUMA protein. In conclusion, Bim and PUMA are two critical mediators of immune cell death in sepsis. Use of a novel cyclodextrin polymer-based, transferrin receptor-targeted siRNA delivery vehicle enables effective administration of antiapoptotic siRNAs to lymphocytes and reverses the immune cell depletion that is a hallmark of this highly lethal disorder.
authors.affiliation: Department of Surgery, Washington University in St Louis, School of Medicine, St Louis, Missouri 63110, USA.
authors.forename: Pavan
authors.initials: P
authors.lastname: Brahmamdam
authors.forename: Eizo
authors.initials: E
authors.lastname: Watanabe
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Unsinger
authors.forename: Katherine C
authors.initials: KC
authors.lastname: Chang
authors.forename: William
authors.initials: W
authors.lastname: Schierding
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Hoekzema
authors.forename: Tony T
authors.initials: TT
authors.lastname: Zhou
authors.forename: Jacquelyn S
authors.initials: JS
authors.lastname: McDonough
authors.forename: Heather
authors.initials: H
authors.lastname: Holemon
authors.forename: Jeremy D
authors.initials: JD
authors.lastname: Heidel
authors.forename: Craig M
authors.initials: CM
authors.lastname: Coopersmith
authors.forename: Jonathan E
authors.initials: JE
authors.lastname: McDunn
authors.forename: Richard S
authors.initials: RS
authors.lastname: Hotchkiss
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics / Q000276:immunology*; D051017:Apoptosis Regulatory Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology; D000072224:Bcl-2-Like Protein 11; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002482:Cellulose / Q000494:pharmacology; D003505:Cyclodextrins / Q000494:pharmacology; D006801:Humans; D008212:Lymphocyte Depletion; D008297:Male; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D051379:Mice; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D034741:RNA, Small Interfering / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D011990:Receptors, Transferrin / Q000819:agonists / Q000235:genetics / Q000276:immunology; D018805:Sepsis / Q000235:genetics / Q000276:immunology*; D014162:Transfection / Q000379:methods*; D025521:Tumor Suppressor Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology
chemical_list: D051017:Apoptosis Regulatory Proteins; C000606775:BCL2L11 protein, human; D000072224:Bcl-2-Like Protein 11; C000606776:Bcl2l11 protein, mouse; D003505:Cyclodextrins; D008565:Membrane Proteins; C488588:PUMA protein, mouse; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; D011990:Receptors, Transferrin; D025521:Tumor Suppressor Proteins; C041953:cyclodextrin polymer; D002482:Cellulose