The Synthetic Antimicrobial Peptide 19-2.5 Interacts with Heparanase and Heparan Sulfate in Murine and Human Sepsis

Abstract

Heparanase is an endo-β-glucuronidase that cleaves heparan sulfate side chains from their proteoglycans. Thereby, heparanase liberates highly potent circulating heparan sulfate-fragments (HS-fragments) and triggers the fatal and excessive inflammatory response in sepsis. As a potential anti-inflammatory agent for sepsis therapy, peptide 19–2.5 belongs to the class of synthetic anti-lipopolysaccharide peptides; however, its activity is not restricted to Gram-negative bacterial infection. We hypothesized that peptide 19–2.5 interacts with heparanase and/or HS, thereby reducing the levels of circulating HS-fragments in murine and human sepsis. Our data indicate that the treatment of septic mice with peptide 19–2.5 compared to untreated control animals lowers levels of plasma heparanase and circulating HS-fragments and reduces heparanase activity. Additionally, mRNA levels of heparanase in heart, liver, lung, kidney and spleen are downregulated in septic mice treated with peptide 19–2.5 compared to untreated control animals. In humans, plasma heparanase level and activity are elevated in septic shock. The ex vivo addition of peptide 19–2.5 to plasma of septic shock patients decreases heparanase activity but not heparanase level. Isothermal titration calorimetry revealed a strong exothermic reaction between peptide 19–2.5 and heparanase and HS-fragments. However, a saturation character has been identified only in the peptide 19–2.5 and HS interaction. In conclusion, the findings of our current study indicate that peptide 19–2.5 interacts with heparanase, which is elevated in murine and human sepsis and consecutively attenuates the generation of circulating HS-fragments in systemic inflammation. Thus, peptide 19–2.5 seems to be a potential anti-inflammatory agent in sepsis.

Immunomodulatory effects of anti-microbial peptides

“…The peptide also blocks a pro-inflammatory response induced by soluble heparan sulfate in serum from patients with gram-negative or gram-positive septic shock. Peptide Pep19-2.5 also interacts with heparanase, an enzyme that is elevated in mammalian sepsis and consecutively decreases the levels of circulating heparan sulfate fragments in systemic inflammation [95]. Due to these activities, peptide Pep19-2.5 was suggested to have a potential for pharmaceutical development as a broadspectrum anti-inflammatory agent in sepsis-induced myocardial inflammation and dysfunction.…”

The synthetic antimicrobial peptide 19-2.5 attenuates septic cardiomyopathy and prevents down-regulation of SERCA2 in polymicrobial sepsis

“…The murine sepsis model was divided into three steps1112: In a first step, a central vein catheter (PE-tube, self-made) was implanted in the jugular vein. Mice were transferred back into the cage to rest for 48 h. The i.v.-line was connected to the syringe pump, and animals received saline (of 100 μl/h) as fluid resuscitation.…”

“…Mice were transferred back into the cage to rest for 48 h. The i.v.-line was connected to the syringe pump, and animals received saline (of 100 μl/h) as fluid resuscitation. In a second step, mice underwent CLP (18-G needle, double puncture) or sham operation1112. Sham mice were not subjected to CLP, but were otherwise treated in the same way.…”

Rol del glicocálix en la Sepsis: Revisión de La literatura y enfoque traslacional

“…En este contexto, estudios sugieren una asociación entre niveles altos de endocano y falla respiratoria aguda [30], sin embargo, en un trabajo reciente realizado por Diego Orbegozo, Lokmane Rahmania [31] (2017), en 96 pacientes encontraron que el endocano en SDRA tiene una baja sensibilidad y moderada especificidad medidos a las 24 horas con un corte de 6 ng/mL y de 14 ng/ml, para predicción de pobre evolución con una curva ROC de 0.66 (0.55-0.74), y no evidenciaron correlación con mortalidad. Todos estos biomarcadores son prometedores, aún es necesario definir en un contexto clínico adecuado, el nivel de corte, y utilidad ya sea como medida pronostica, diagnostica u objetivo de reanimación.…”

“…Endocan curva ROC para predicción de mala evolución; C, Diagrama de cajas, Endocan en sobrevivientes y no sobrevivientes [31]. …”