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Proteomic Evidence for G-CSF Induced Fibrinolysis and Neovascularization

Darwin Eton*, Amelia Bartholomew, Guolin Zhou, Tong-Chuan He

Objective: To verify proteomic evidence for fibrinolysis and Neovascularization (NV) in patients with Chronic Limb Threatening Ischemia (CLTI) treated with a novel cell therapy approach.

Methods: Six CLTI patients were treated for one month with filgrastim (Amgen Inc) 10 mcg/kg SQ every 72 hours, and an external Programmed Infra-Geniculate Compression Pump (PCP) for 3 hours a day. Blood was drawn on day 1 (baseline), and on days 15 and 30 (24 hours after the 5th and 10th Filgrastim doses). On each of these days blood was sampled before and after 2 hours of supervised PCP. Serum ELISA was used to measure the concentration of Plasmin, Fibrin Degradation Products (FDP), VEGF-A, Hepatocyte Growth Factor (HGF) and MMP-9.

Results: Statistically significant elevation of plasmin and FDP (p<0.001) were consistently measured 24 hours after each Filgrastim administration and was not influenced by the pump. No hemorrhagic complications occurred. Pump independent significant elevation in VEGF-A, HGF and MMP-9, each associated with NV, was also observed.

Conclusion: These observations support angiographic, hemodynamic and clinical evidence of fibrinolysis and neovascularization achieved with this novel therapy. These data support further clinical testing of this cell therapy in CLTI, as well as in other ischemic tissue beds.