PMID- 25560976
OWN - NLM
STAT- MEDLINE
DCOM- 20151106
LR  - 20181202
IS  - 1520-4812 (Electronic)
IS  - 1043-1802 (Linking)
VI  - 26
IP  - 2
DP  - 2015 Feb 18
TI  - DC3-decorated polyplexes for targeted gene delivery into dendritic cells.
PG  - 213-24
LID - 10.1021/bc500529d [doi]
AB  - Dendritic cells (DCs) are a family of specialized antigen presenting cells (APCs) 
      that detect antigens and initiate a wide spectrum of immune responses against 
      them. These characteristics make them promising candidates for immunotherapy 
      manipulations. In this study we designed and synthesized DC-targeted block 
      copolymers composed of linear polyethylenimine (PEI) conjugated to hydrophilic 
      polyethylene glycol (PEG) installed with a DC-targeting peptide (DC3, primary 
      sequence FYPSYHSTPQRP). Two different conjugation procedures (basic and modified) 
      were employed to synthesize the DC3-PEG-b-PEI and the control SCRM-PEG-b-PEI 
      (with a scrambled DC3 peptide sequence) by one-pot synthesis, in two steps. In 
      the first, basic conjugation procedure, PEG with N-hydroxysuccinimide (NHS) ester 
      and maleimide (MAL) groups (NHS-PEG-MAL, 3.5 kDa) was first coupled to linear PEI 
      (25 kDa) via the NHS group to yield the intermediate MAL-PEG-b-PEI, that was then 
      conjugated to N-terminus-cysteine harboring peptides DC3 or SCRM via the MAL 
      double bond to yield the final DC3-PEG-b-PEI or SCRM-PEG-b-PEI copolymers, 
      respectively. In the second, modified conjugation procedure, Fmoc-cysteine 
      harboring DC3 or SCRM peptides were first conjugated to NHS-PEG-MAL via the MAL 
      group followed by coupling to linear PEI via the NHS functional group. Fmoc 
      cleavage yielded the same final product as in the basic procedure. The modified 
      conjugation procedure was capable of yielding block copolymers richer with 
      peptides, as determined by (1)H NMR analysis. Self-assembly of DC3-PEG-b-PEI 
      copolymers and DNA molecules yielded nanosized polyion complexes (polyplexes), 
      with lower surface charge and limited cytotoxicity when compared to the PEI 
      building block. Significant transfection efficiency of the DC-targeted polyplexes 
      by murine dendritic DC2.4 cells was observed only in DC3-PEG-b-PEI/DNA polyplexes 
      synthesized by the modified conjugation procedure. These polyplexes, with higher 
      peptide-load, showed greater transfection capability in DC2.4 cells relative to 
      the control nontargeted SCRM-PEG-b-PEI/DNA polyplexes, but not in endothelial 
      cells. The transfection efficiency was comparable to or higher than that of the 
      PEI/DNA positive control. The results indicate that PEGylated-PEI polyplexes show 
      significant transfection efficiency into DCs when decorated with DC3 peptide, and 
      are attractive candidates for immunotherapy via DCs.
FAU - Golani-Armon, Adi
AU  - Golani-Armon A
AD  - Department of Clinical Biochemistry and Pharmacology, Faculty of Health Sciences, 
      and double daggerIlse Katz Institute for Nanoscale Science and Technology, Ben-Gurion 
      University of the Negev , Beer-Sheva, Israel 84105.
FAU - Golan, Moran
AU  - Golan M
FAU - Shamay, Yosi
AU  - Shamay Y
FAU - Raviv, Lior
AU  - Raviv L
FAU - David, Ayelet
AU  - David A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150209
PL  - United States
TA  - Bioconjug Chem
JT  - Bioconjugate chemistry
JID - 9010319
RN  - 0 (Maleimides)
RN  - 0 (Peptides)
RN  - 0 (Succinimides)
RN  - 2519R1UGP8 (maleimide)
RN  - 3WJQ0SDW1A (Polyethylene Glycols)
RN  - 9002-98-6 (Polyethyleneimine)
RN  - 9007-49-2 (DNA)
RN  - MJE3791M4T (N-hydroxysuccinimide)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Cell Line
MH  - DNA/*administration & dosage/genetics
MH  - Dendritic Cells/*metabolism
MH  - Maleimides/chemistry/metabolism
MH  - Mice
MH  - Peptides/*chemistry/metabolism
MH  - Polyethylene Glycols/*chemistry/metabolism
MH  - Polyethyleneimine/*chemistry/metabolism
MH  - Succinimides/chemistry/metabolism
MH  - *Transfection
EDAT- 2015/01/07 06:00
MHDA- 2015/11/07 06:00
CRDT- 2015/01/07 06:00
PHST- 2015/01/07 06:00 [entrez]
PHST- 2015/01/07 06:00 [pubmed]
PHST- 2015/11/07 06:00 [medline]
AID - 10.1021/bc500529d [doi]
PST - ppublish
SO  - Bioconjug Chem. 2015 Feb 18;26(2):213-24. doi: 10.1021/bc500529d. Epub 2015 Feb 
      9.