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Updated: Jun 22, 2025
Murine Model of Hindlimb Ischemia
Published on: January 21, 2009
Loss of c-Kit in Endothelial Cells Protects against Hindlimb Ischemia
Gustavo Falero-Diaz1, Catarina de A Barboza2, Roberto I Vazquez-Padron1
1Department of Surgery, Miller School of Medicine, University of Miami, 1600 NW 10th Ave, RMSB, Miami, FL 33136, USA.
Insights
Endothelial SCF/c-Kit signaling impairs blood vessel formation (arteriogenesis) and weakens the endothelial barrier in critical limb ischemia. Reducing this signaling improves limb perfusion and barrier integrity.
Area of Science:
- Vascular Biology
- Molecular Medicine
- Regenerative Medicine
Background:
- Critical limb ischemia (CLI) is a severe stage of peripheral artery disease (PAD) with limited treatment options.
- While c-Kit has shown angiogenic potential, its specific role in endothelial cells during ischemia is unclear.
- Understanding endothelial c-Kit signaling is crucial for developing new CLI therapies.
Purpose of the Study:
- To investigate the role of endothelial c-Kit and its ligand, SCF, in arteriogenesis and endothelial barrier function during hindlimb ischemia.
- To determine the impact of specifically reducing endothelial c-Kit or SCF on blood flow recovery and vascular integrity.
Main Methods:
- Generated conditional knockout mice with reduced endothelial c-Kit or SCF using VE-Cadherin CreERT2 system.
- Induced hindlimb ischemia to assess limb perfusion and arteriogenesis.
- Utilized aortic crush model to evaluate endothelial barrier permeability.
Main Results:
- Confirmed reduced c-Kit and SCF expression in endothelial cells of knockout mice.
- Observed improved limb perfusion and arteriogenesis in mice with reduced endothelial c-Kit or SCF.
- Demonstrated preserved endothelial barrier integrity in knockout mice after aortic injury.
Conclusions:
- Endothelial SCF/c-Kit signaling has detrimental effects on arteriogenesis.
- This signaling pathway also compromises endothelial barrier integrity.
- Targeting endothelial SCF/c-Kit may offer a therapeutic strategy for CLI.
Background:
Critical limb ischemia (CLI) is the end stage of peripheral artery disease (PAD), and around 30% of CLI patients are ineligible for current treatments. The angiogenic benefits of c-Kit have been reported in the ischemia scenario; however, the present study demonstrates the effects of specific endothelial c-Kit signaling in arteriogenesis during hindlimb ischemia.
Methods:
We created conditional knockout mouse models that decrease c-Kit (c-Kit VE-Cadherin CreERT2-c-Kit) or its ligand (SCF VE-Cadherin CreERT2-SCF) specifically in endothelial cells (ECs) after tamoxifen treatment. These mice and a control group (wild-type VE-Cadherin CreERT2-WT) were subjected to hindlimb ischemia or aortic crush to evaluate perfusion/arteriogenesis and endothelial barrier permeability, respectively.
Results:
Our data confirmed the lower gene expression of c-Kit and SCF in the ECs of c-Kit and SCF mice, respectively. In addition, we confirmed the lower percentage of ECs positive for c-Kit in c-Kit mice. Further, we found that c-Kit and SCF mice had better limb perfusion and arteriogenesis compared to WT mice. We also demonstrated that c-Kit and SCF mice had a preserved endothelial barrier after aortic crush compared to WT.
Conclusions:
Our data demonstrate the deleterious effects of endothelial SCF/c-Kit signaling on arteriogenesis and endothelial barrier integrity.
