Targeted delivery of siRNA to cell death proteins in sepsis.
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.
Brahmamdam
Pavan
P
Department of Surgery, Washington University in St Louis, School of Medicine, St Louis, Missouri 63110, USA.
Watanabe
Eizo
E
Unsinger
Jacqueline
J
Chang
Katherine C
KC
Schierding
William
W
Hoekzema
Andrew S
AS
Zhou
Tony T
TT
McDonough
Jacquelyn S
JS
Holemon
Heather
H
Heidel
Jeremy D
JD
Coopersmith
Craig M
CM
McDunn
Jonathan E
JE
Hotchkiss
Richard S
RS
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
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