Abstract 320: Delivery of Hepatocyte Growth Factor mRNA From Nanofibrillar Scaffolds for Treatment of Peripheral Arterial Disease

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Ngan F Huang ◽  
Luqia Hou ◽  
Cynthia Alcazar ◽  
Zachary Strassberg ◽  
Michael Hopkins ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Growth Factor ◽  
Transfection Efficiency ◽  
Blood Perfusion ◽  
Arterial Disease ◽  
Tissue Culture Plastic ◽  
Gfp Expression ◽  
Peripheral Arterial ◽  
Hepatocyte Growth ◽  
Scaffold Group

Biological approaches to augment angiogenesis are promising for treatment of peripheral arterial disease (PAD). We propose the use of scaffold-based modified mRNA (mmRNA) delivery as a favorable approach for transient, localized gene delivery. We hypothesized that hepatocyte growth factor (HGF) mmRNA-seeded nanofibrillar scaffolds will enable localized and temporally controlled delivery of mmRNA, leading to augmentation of angiogenesis in a murine model of PAD. To establish the efficacy of mmRNA therapy, mmRNA encoding green fluorescence protein (GFP) was used as a fluorescent reporter for quantification of transfection efficiency. Aligned nanofibrillar collagen scaffolds were loaded with mmRNA and lipofectamine transfection agent. The temporal kinetics of mmRNA release into media was measured by ribogreen assay. To determine the transfection efficiency, human fibroblasts were cultured on the aligned nanofibrillar scaffolds, or on tissue culture plastic, and the efficiency of transfection was measured for up to 7 days and assayed for GFP expression. Based on ribogreen assay, the cumulative release of GFP mmRNA over the course of 14 days was 235 ng/cm scaffold. In vitro transfection efficiency on aligned scaffolds (75%) was markedly higher than on tissue culture plastic (45%) after 24h. The persistence of cellular transfection as quantified by western blotting showed GFP expression >5 days post-transfection. Next, to demonstrate therapeutic efficacy for treatment of PAD, scaffolds releasing HGF or GFP mmRNA were transplanted to the site of the murine ischemic hindlimb. At the end of the 14 day experiment, laser Doppler spectroscopy showed that HGF mmRNA scaffold group had a higher mean perfusion ratio (0.32 ±0.10) than the GFP mmRNA scaffold group (0.23±0.14), suggesting that HGF-scaffolds improved blood perfusion. In summary, these data suggest that HGF mmRNA-releasing scaffolds marked improved blood perfusion in a murine model of PAD.

scholarly journals Safety Evaluation of Clinical Gene Therapy Using Hepatocyte Growth Factor to Treat Peripheral Arterial Disease

Hypertension ◽  
2004 ◽  
Vol 44 (2) ◽  
pp. 203-209 ◽  
Author(s):  
Ryuichi Morishita ◽  
Motokuni Aoki ◽  
Naotaka Hashiya ◽  
Hirofumi Makino ◽  
Keita Yamasaki ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Growth Factor ◽  
Peripheral Arterial Disease ◽  
Hepatocyte Growth Factor ◽  
Safety Evaluation ◽  
Arterial Disease ◽  
Clinical Gene Therapy ◽  
Peripheral Arterial ◽  
Hepatocyte Growth

Potential Role of Hepatocyte Growth Factor, a Novel Angiogenic Growth Factor, in Peripheral Arterial Disease

Circulation ◽  
1999 ◽  
Vol 100 (suppl_2) ◽  
Author(s):  
Shin-ichiro Hayashi ◽  
Ryuichi Morishita ◽  
Shigefumi Nakamura ◽  
Kei Yamamoto ◽  
Atsushi Moriguchi ◽  
...  
Keyword(s):  
Growth Factor ◽  
Peripheral Arterial Disease ◽  
Hepatocyte Growth Factor ◽  
Hind Limb ◽  
Arterial Disease ◽  
Angiogenic Growth Factor ◽  
Peripheral Arterial ◽  
Hepatocyte Growth ◽  
Hypoxic Treatment

Background —Although hepatocyte growth factor (HGF), a novel angiogenic growth factor, plays an important role in angiogenesis, regulation of local HGF production under hypoxia has not yet been clarified in vascular smooth muscle cells (VSMC) and endothelial cells (EC). Thus, we have studied the role of HGF in hypoxia-induced endothelial injury and the regulation of local vascular HGF expression in response to hypoxia. Methods and Results —HGF attenuated hypoxia-induced endothelial cell death. Importantly, hypoxic treatment of EC resulted in a significant decrease in local HGF production according to the severity of hypoxia and increased VEGF. Similarly, hypoxia significantly decreased in mRNA and protein of HGF and increased VEGF production in VSMC. In organ culture system, local HGF production was also significantly decreased by hypoxia ( P <0.01). Downregulation of HGF by hypoxia is due to a significant decrease in cAMP, as hypoxic treatment decreased cAMP, a stimulator of HGF. Although active TGF-β, a suppressor of HGF, was increased at 72 hours after hypoxic treatment, treatment of anti-TGF-β antibody did not attenuate decreased HGF production. Finally, rHGF was intra-arterially administered into unilateral hind limb ischemia rabbits, to evaluate in vivo angiogenic activity. Of importance, a single intra-arterial administration of rHGF reduced severe necrosis due to ischemia in rabbit muscle, accompanied by a significant increase in angiographic score ( P <0.01). Conclusions —Overall, these data demonstrated that hypoxic treatment of vascular cells significantly downregulated HGF production due to decreased cAMP, suggesting their potential roles in the pathophysiology of ischemic diseases. Moreover, administration of rHGF induced therapeutic angiogenesis, accompanied by improvement of necrotic changes in the ischemic hind limb model, as cytokine supplement therapy for peripheral arterial disease.

scholarly journals Association of Circulating Hepatocyte Growth Factor and Risk of Incident Peripheral Artery Disease: The Multi-Ethnic Study of Atherosclerosis

Angiology ◽  
2020 ◽  
Vol 71 (6) ◽  
pp. 544-551
Author(s):  
Parveen K. Garg ◽  
Petra Buzkova ◽  
Christina L. Wassell ◽  
Matthew Allison ◽  
Michael Criqui ◽  
...  
Keyword(s):  
Growth Factor ◽  
Hepatocyte Growth Factor ◽  
Ankle Brachial Index ◽  
Arterial Disease ◽  
Ethnic Study ◽  
Increased Risk ◽  
Artery Disease ◽  
Peripheral Arterial ◽  
Hepatocyte Growth

Higher levels of hepatocyte growth factor (HGF) have been associated with the presence of peripheral arterial disease (PAD), but prospective associations are unknown. We examined the association of circulating HGF levels with incident PAD. Between 2000 and 2002, HGF was measured in 6742 Multi-Ethnic Study of Atherosclerosis participants without PAD. Incident clinical PAD, adjudicated on the basis of a positive history for the presence of disease-related symptoms or treatment, was ascertained through 2015. Incident low ankle-brachial index (ABI), defined as an ABI < 0.9 and a decline of ≥ 0.15, was assessed among 5736 individuals who had an ABI > 0.9 at baseline and ≥1 follow-up ABI measurement 3 to 10 years later. There were 116 clinical PAD and 197 low ABI events that occurred over a median follow-up of 14 and 9 years, respectively. After adjustment for demographic and clinical variables, a standard deviation increment of HGF (303 ng/L) was associated with an increased risk of clinical PAD (hazard ratio: 1.21; 95% confidence interval [CI]: 1.05-1.39) but not a low ABI (rate ratio: 1.03; 95% CI: 0.85-1.25). In conclusion, higher HGF levels were modestly associated with an increased risk of developing clinical PAD.

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