Efficient gene delivery to pancreatic islets with ultrasonic microbubble destruction technology.
This study describes a method of gene delivery to pancreatic islets of adult, living animals by ultrasound targeted microbubble destruction (UTMD). The technique involves incorporation of plasmids into the phospholipid shell of gas-filled microbubbles, which are then infused into rats and destroyed within the pancreatic microcirculation with ultrasound. Specific delivery of genes to islet beta cells by UTMD was achieved by using a plasmid containing a rat insulin 1 promoter (RIP), and reporter gene expression was regulated appropriately by glucose in animals that received a RIP-luciferase plasmid. To demonstrate biological efficacy, we used UTMD to deliver RIP-human insulin and RIP-hexokinase I plasmids to islets of adult rats. Delivery of the former plasmid resulted in clear increases in circulating human C-peptide and decreased blood glucose levels, whereas delivery of the latter plasmid resulted in a clear increase in hexokinase I protein expression in islets, increased insulin levels in blood, and decreased circulating glucose levels. We conclude that UTMD allows relatively noninvasive delivery of genes to pancreatic islets with an efficiency sufficient to modulate beta cell function in adult animals.
Chen
Shuyuan
S
Department of Internal Medicine, Cardiology Section, Baylor University Medical Center, Baylor Heart and Vascular Institute, 621 North Hall Street, Suite H030, Dallas, TX 75226, USA.
Ding
Jia-huan
JH
Bekeredjian
Raffi
R
Yang
Bing-zhi
BZ
Shohet
Ralph V
RV
Johnston
Stephen A
SA
Hohmeier
Hans E
HE
Newgard
Christopher B
CB
Grayburn
Paul A
PA
D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D002096:C-Peptide / Q000097:blood; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D006593:Hexokinase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000235:genetics / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D045423:Microbubbles; D018613:Microscopy, Confocal; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014465:Ultrasonics; D000097573:Red Fluorescent Protein
D001786:Blood Glucose; D002096:C-Peptide; D006593:Hexokinase; D007328:Insulin; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein