<h3>Purpose/Objective(s)</h3> Radiation induced toxicity (RIT) has long been a notorious limit of radiotherapy. RIT can only be circumvented by patient selection or dose de-escalation. No effective treatment of RIT is available. Endothelium injury (EnI) has been recognized as the key pathogenesis in radiotoxicity. EnI initiates the vicious cascade and propagates to various RIT. Defibrotide (DF), a mixture of oligonucleotides, is a potent endothelium protector and is the only medication approved for the treatment of post-HSCT severe hepatic sinusoid obstructive syndrome (SOS), with survival benefits and minimal toxicity. The protective effect of DF has been exhibited in chemo-toxicity, hypoxia, physical injury and infection, but not yet in radiation (RT). We hypothesize that DF alleviates RIT through a similar mechanism. Here, we present the first report and primitive results of an in vitro study of DF in RT setting. <h3>Materials/Methods</h3> Primary human hepatic sinusoidal endothelial cells (HHSEC) and human umbilical vein endothelial cells (HUVEC) were cultured and RT for a single 4Gy or 8Gy with or without DF at the final concentration of 300ug/ml. Assays for cell viability, survival and proliferation were evaluated. Molecule expression pattern was analyzed by rt-PCR. All the assays were replicated 3 times. <h3>Results</h3> After RT, HHSECs became sparse and deformed under the microscope. In contrast, HHSECs cultured with DF before RT generally appeared normal. In accordance, a cell counting kit assay showed that viability of HHSECs was preserved after RT at the presence of DF, while it was dramatically reduced by RT alone (p<0.001). Flow cytometry analysis of programed cell death 48h post-RT confirmed that DF significantly reduced both early and late apoptosis (8.9% vs 14.6%, p<0.05). In the proliferation assay, RT alone almost quartered the number of HUVEC clones, while DF co-treatment partially prevented the extermination. DF alone had no deleterious effect compared with control groups, in concordance with published studies. RT-PCR revealed elevated expression of pro-inflammatory and pro-coagulant molecules. In detail, raised mRNA levels of von Willebrand factor (vWF), ICAM-1, VCAM-1, IL-1a, IL-1b, IL-6, TNF-a and eNOS were detected after RT, which were down regulated by DF. DNA break assay by γ-H2AX formation was performed, which showed no drastic difference of γ-H2AX staining after RT with or without DF, implicating other mechanisms. Besides, DF prophylaxis before RT rather than salvage after RT revealed better endothelium protection. We then built a rat model of radiation induced endothelium injury (RIEI) by whole liver exposure. However, due to the COVID-19 outbreak and temporary un-availability of DF, further study is in waiting. <h3>Conclusion</h3> Effective therapy of RIT is a huge unmet medical need. DF, an approved, well-tolerated orphan drug for post-HSCT SOS, exhibited definitive protective effect in RIEI as well. Further studies, not only pre-clinically but also in clinical scenarios are warranted and in eager expectation.
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