PMID- 12865820
OWN - NLM
STAT- MEDLINE
DCOM- 20030904
LR  - 20161124
IS  - 0041-1337 (Print)
IS  - 0041-1337 (Linking)
VI  - 76
IP  - 1
DP  - 2003 Jul 15
TI  - Double genetic modification of adenovirus fiber with RGD polylysine motifs 
      significantly enhances gene transfer to isolated human pancreatic islets.
PG  - 252-61
AB  - BACKGROUND: New strategies for improving durable functional islet mass will be 
      instrumental in facilitating islet transplantation as a cure for type 1 diabetes 
      mellitus. The ability to transfer immunoregulatory or cytoprotective genes into 
      pancreatic islets may enhance survival. Adenoviral vectors (Ad5) have been used 
      widely to deliver therapeutic genes to different tissues. Limitations associated 
      with the use of Ad5 for gene therapy are related to the reliance of the virus on 
      the presence of its primary receptor, the transient nature of the transgene 
      expression, and the immediate inflammatory and immune response elicited by the 
      infection. Because the arginine-glycine-aspartame (RGD) and polylysine (pK7) 
      motifs have been shown to enhance Ad5 infection through an Ad5 
      receptor-independent pathway, we hypothesized that they could act additively to 
      improve infectivity and reduce toxicity to isolated human pancreatic islets 
      (IHPI). METHODS: Hand-picked IHPI were infected with nonmodified Ad5, 
      single-modified Ad5 with RGD (Ad5RGD) or pK7 (ad5pK7), and Ad5RGDpK7. 
      Transfection efficiency was evaluated by green fluorescent protein and luciferase 
      expression. Apoptosis was assessed using a quantitative assay, activation of 
      caspase 3 by a colorimetric assay, nuclear factor (NF)-kappaB nuclear 
      translocation using a promoter-luciferase NF-kappaB responsive construct, 
      regulated on activation normal T-cell expressed and secreted (RANTES) by 
      enzyme-linked immunosorbent assay. In vivo functionality was evaluated after 
      transplantation into diabetic nonobese diabetic severe combined immunodeficiency 
      mice. RESULTS: Compared with unmodified and singly-modified Ad5 vectors, 
      Ad5RGDpK7 demonstrated the highest infectivity. After the infection of IHPI with 
      adenoviral vectors using the minimal dose required to infect greater than 80% of 
      the islet cells (Ad5, 500 viral particles [VP]/cell; Ad5RGD and Ad5pK7, 10 
      VP/cell; Ad5RGDpK7, 0.1 VP/cell), islets infected with Ad5RGDpK7 presented a 
      significant reduction in apoptosis, NF-kappaB nuclear translocation, RANTES 
      expression, and higher glucose disposal rate; reduced Ad5-driven specific Th1 and 
      antibody response were also observed. CONCLUSIONS: Ad5RGDpK7 exhibited higher 
      transfection efficiency, allowing a significant reduction in the viral dose 
      required to infect greater than 80% of the islet cells. The reduction in the 
      viral dose was associated with reduced toxicity, inflammation, and immune 
      responses related to Ad5 infection. This strategy may thus be used to 
      successfully modify isolated pancreatic islets.
FAU - Contreras, Juan L
AU  - Contreras JL
AD  - Department of Surgery, University of Alabama at Birmingham, Birmingham, AL 35294, 
      USA. juan.contreras@ccc.uab.edu.
FAU - Wu, Hongju
AU  - Wu H
FAU - Smyth, Cheryl A
AU  - Smyth CA
FAU - Eckstein, Christopher P
AU  - Eckstein CP
FAU - Young, Carlton J
AU  - Young CJ
FAU - Seki, Toshiro
AU  - Seki T
FAU - Bilbao, Guadalupe
AU  - Bilbao G
FAU - Curiel, David T
AU  - Curiel DT
FAU - Eckhoff, Devin E
AU  - Eckhoff DE
LA  - eng
GR  - P50 CA89019/CA/NCI NIH HHS/United States
GR  - R01 CA86881/CA/NCI NIH HHS/United States
GR  - R01 HL67962/HL/NHLBI NIH HHS/United States
GR  - U19 DK57958/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Transplantation
JT  - Transplantation
JID - 0132144
RN  - 0 (Oligopeptides)
RN  - 25104-18-1 (Polylysine)
RN  - 78VO7F77PN (arginyl-glycyl-aspartic acid)
SB  - IM
MH  - Adenoviridae/genetics/*physiology
MH  - Animals
MH  - Cadaver
MH  - Diabetes Mellitus, Experimental/surgery
MH  - Gene Transfer Techniques
MH  - *Genetic Vectors
MH  - Humans
MH  - Islets of Langerhans/*physiology/virology
MH  - Islets of Langerhans Transplantation/*methods
MH  - Male
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Mice, Inbred NOD
MH  - Mice, SCID
MH  - Oligopeptides/*pharmacology
MH  - Polylysine/*pharmacology
MH  - Tissue Donors
MH  - Transplantation, Heterologous/*methods
EDAT- 2003/07/17 05:00
MHDA- 2003/09/05 05:00
CRDT- 2003/07/17 05:00
PHST- 2003/07/17 05:00 [pubmed]
PHST- 2003/09/05 05:00 [medline]
PHST- 2003/07/17 05:00 [entrez]
AID - 10.1097/01.TP.0000066361.02042.CA [doi]
PST - ppublish
SO  - Transplantation. 2003 Jul 15;76(1):252-61. doi: 
      10.1097/01.TP.0000066361.02042.CA.