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Canadian Journal of Cardiology

Congenital Absence of Nitric Oxide Synthase 3 Potentiates Cardiac Dysfunction and Reduces Survival in Doxorubicin- and Trastuzumab-Mediated Cardiomyopathy

Published:November 18, 2013DOI:https://doi.org/10.1016/j.cjca.2013.11.013

      Abstract

      Background

      Doxorubicin (DOX) and trastuzumab (TRZ) are highly effective chemotherapeutic agents in the breast cancer setting, limited by their cardiotoxic side effects. Among the potential mechanisms for this drug-induced cardiomyopathy, increased production of oxidative stress (OS) through a nitric oxide synthase 3 (NOS3)-dependent pathway has gained recent attention. The objective of the study was to determine the role of NOS3 and OS in a clinically relevant female murine model of DOX- and TRZ-induced heart failure.

      Methods

      A total of 120 female mice (60 wild-type [WT] and 60 NOS3 knockout [NOS3−/−]) were treated with either 0.9% saline, DOX, TRZ, or DOX with TRZ (DOX+TRZ). Serial echocardiography was performed for a total of 10 days, after which the mice were euthanized for histological and biochemical analyses.

      Results

      In WT female mice receiving DOX+TRZ, left ventricular ejection fraction (LVEF) decreased from 75 ± 3% at baseline to 46 ± 2% at day 10 (P < 0.05). In the NOS3−/− group, LVEF decreased from 72 ± 3% at baseline to 35 ± 2% at day 10 (P < 0.05). LVEF was significantly lower in NOS3−/− female mice receiving DOX+TRZ than WT mice at day 10 (P < 0.05). Compared with WT, NOS3−/− female mice also demonstrated increased mortality after treatment with DOX+TRZ, corroborating the echocardiographic findings. Histological analysis demonstrated increased myofibrillar degradation and loss of cell integrity in NOS3−/− female mice treated with DOX+TRZ. There was increased generation of oxidized phosphatidylcholine, a marker of OS, in NOS3−/− female mice receiving DOX+TRZ compared with control mice.

      Conclusions

      Congenital absence of NOS3 potentiates the cardiotoxic side effects of DOX+TRZ in an acute female murine model of chemotherapy-induced cardiomyopathy.

      Résumé

      Introduction

      La doxorubicine (DOX) et le trastuzumabe (TRZ) sont des agents chimiothérapeutiques très efficaces contre le cancer du sein, qui sont limités par leurs effets secondaires cardiotoxiques. Parmi les mécanismes potentiels de cette cardiomyopathie induite par le médicament, l’augmentation de la production du stress oxydatif (SO) par une voie dépendante du monoxyde d’azote synthase 3 (NOS3) a récemment attiré l’attention. L’objectif de l’étude était de déterminer le rôle du NOS3 et du SO par un modèle murin femelle d’insuffisance cardiaque induite par la DOX et le TRZ.

      Méthodes

      Un total de 120 souris femelles (60 de type sauvage [TS] et 60 invalidées pour le NOS3 ont été traitées soit par le soluté physiologique solution salée à 0,9 %, la DOX ou le TRZ, ou la DOX et le TRZ [DOX+TRZ]. L’échocardiographie sérielle a été réalisée durant 10 jours après lesquels les souris ont été euthanasiées pour procéder aux analyses histologiques et biochimiques.

      Résultats

      Chez les souris femelles de TS traitées par DOX+TRZ, la fraction d’éjection ventriculaire gauche [FEVG] a diminué de 75 ± 3 % au début à 46 ± 2 % au 10e jour [P < 0,05]. Dans le groupe NOS3-/-, la FEVG a diminué de 72 ± 3 % au début à 35 ± 2 % au 10e jour (P < 0,05). La FEVG a été significativement plus basse chez les souris femelles NOS3-/- traitées par DOX+TRZ que chez les souris de TS au 10e jour (P < 0,05). Comparativement aux souris de TS, les souris femelles NOS3-/- ont également démontré une augmentation de la mortalité après le traitement par DOX+TRZ, ce qui corrobore les résultats de l’échocardiographie. Les analyses histologiques ont démontré une augmentation de la dégradation de la myofibrille et la perte de l’intégrité de la cellule chez les souris femelles NOS3-/- traitées par DOX+TRZ. Il y a eu augmentation de la production de phosphatidylcholine oxydée, un marqueur du SO, chez les souris femelles NOS3-/- traitées par DOX+TRZ comparativement aux souris témoins.

      Conclusions

      L’absence congénitale de NOS3 potentialise les effets secondaires cardiotoxiques du traitement par DOX+TRZ dans le modèle murin femelle de cardiomyopathie induite par la chimiothérapie à court terme.
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