The role of cell-free DNA in fibrinolysis for intraventricular hemorrhage

Fangke Xie MD1, Qiang Tan MD2, Anyong Yu MD, PhD1, Peiwen Guo MD2, Ling Wang MD2, Zongwei Zeng MD2, Liang Liang MD2, Jishu Xian MD2, Hua Feng MD, PhD2, and Zhi Chen MD, PhD2
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  • 1 Department of Emergency, Affiliated Hospital of Zunyi Medical University, Zunyi; and
  • | 2 Department of Neurosurgery, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, China
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OBJECTIVE

Tissue plasminogen activator (tPA) fibrinolysis did not improve functional outcomes of patients with intraventricular hemorrhage (IVH), largely because of the unsatisfactory clot clearance. The presence of neutrophil extracellular traps (NETs) within the clot has been confirmed to impair tPA fibrinolysis, but the mechanism has been unclear. The authors hypothesized that cell-free DNA (cfDNA), the main framework of NETs, might be the important reason for the fibrinolysis resistance, and they validated the hypothesis, hoping to provide a new target to promote intraventricular fibrinolysis.

METHODS

First, cfDNA was detected in IVH clots by immunofluorescence staining in a rat model of IVH. Second, after blood (with or without exogenous cfDNA) intraventricular injection, IVH rats were given intraventricular infusion of 2 μl of saline, tPA, or tPA + DNase1 randomly. Then, the ventricular volume, animal behavior, and reactive astrocyte proliferation were assessed. Third, the IVH clots were collected for fibrinolysis assay in vitro. Finally, the effects of exogenous cfDNA in IVH were evaluated.

RESULTS

The presence of cfDNA in clots was observed as early as 1 hour after IVH. Compared with the whole-blood model, blood + cfDNA caused more severe ventricular dilation (day 7: blood 32.47 ± 2.096 mm3 vs blood + DNA 40.09 ± 2.787 mm3, p < 0.05), increased fibrinolysis resistance to tPA (day 7: tPA + DNA 26.04 ± 1.318 mm3 vs tPA 22.15 ± 1.706 mm3, p < 0.05), and further deteriorated the functional defects in rats (blood vs blood + DNA, p < 0.05). Degradation of cfDNA by DNase1 further enhanced the fibrinolysis effects on relieving the ventricular dilation (day 7: tPA + DNase1 11.67 ± 2.023 mm3 vs tPA, p < 0.05), improving the functional outcome (tPA vs tPA + DNase1, p < 0.05) and reducing periventricular astrocyte proliferation.

CONCLUSIONS

cfDNA impaired tPA fibrinolysis for IVH, and degradation of cfDNA may be a new target to improve this condition.

ABBREVIATIONS

cfDNA = cell-free DNA; DAPI = 4′,6-diamidino-2-phenylindole; EVD = external ventricular drainage; GFAP = glial fibrillary acidic protein; H3Cit = citrullinated histone 3; ICP = intracranial pressure; IVH = intraventricular hemorrhage; mNSS = modified Neurological Severity Score; NET = neutrophil extracellular trap; tPA = tissue plasminogen activator.

Illustration from Kim et al. (pp 1164–1172). Copyright Eui Hyun Kim. Published with permission.

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