Apaf-1 inhibitors protect from unwanted cell death in in vivo models of kidney ischemia and chemotherapy induced ototoxicity

PLoS One. 2014 Oct 20;9(10):e110979. doi: 10.1371/journal.pone.0110979. eCollection 2014.

Abstract

Background: Excessive apoptosis induces unwanted cell death and promotes pathological conditions. Drug discovery efforts aimed at decreasing apoptotic damage initially targeted the inhibition of effector caspases. Although such inhibitors were effective, safety problems led to slow pharmacological development. Therefore, apoptosis inhibition is still considered an unmet medical need.

Methodology and principal findings: The interaction between Apaf-1 and the inhibitors was confirmed by NMR. Target specificity was evaluated in cellular models by siRNa based approaches. Cell recovery was confirmed by MTT, clonogenicity and flow cytometry assays. The efficiency of the compounds as antiapoptotic agents was tested in cellular and in vivo models of protection upon cisplatin induced ototoxicity in a zebrafish model and from hypoxia and reperfusion kidney damage in a rat model of hot ischemia.

Conclusions: Apaf-1 inhibitors decreased Cytc release and apoptosome-mediated activation of procaspase-9 preventing cell and tissue damage in ex vivo experiments and in vivo animal models of apoptotic damage. Our results provide evidence that Apaf-1 pharmacological inhibition has therapeutic potential for the treatment of apoptosis-related diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / adverse effects*
  • Antineoplastic Agents / pharmacology
  • Apoptosis
  • Apoptotic Protease-Activating Factor 1 / antagonists & inhibitors*
  • Apoptotic Protease-Activating Factor 1 / metabolism
  • Cell Death / drug effects
  • Cisplatin / adverse effects*
  • Cisplatin / pharmacology
  • Disease Models, Animal
  • HeLa Cells
  • Hearing Loss* / chemically induced
  • Hearing Loss* / metabolism
  • Hearing Loss* / pathology
  • Heterocyclic Compounds, 4 or More Rings / chemistry
  • Heterocyclic Compounds, 4 or More Rings / pharmacology*
  • Humans
  • Kidney / metabolism
  • Kidney / pathology
  • Kidney Diseases / metabolism*
  • Kidney Diseases / pathology
  • Male
  • Mice
  • Rats
  • Reperfusion Injury / metabolism*
  • Reperfusion Injury / pathology
  • Zebrafish / metabolism*
  • Zebrafish Proteins / antagonists & inhibitors*
  • Zebrafish Proteins / metabolism

Substances

  • APAF1 protein, human
  • Antineoplastic Agents
  • Apaf1 protein, mouse
  • Apaf1 protein, rat
  • Apoptotic Protease-Activating Factor 1
  • Heterocyclic Compounds, 4 or More Rings
  • Zebrafish Proteins
  • Cisplatin

Grants and funding

This work was supported by grants from the Spanish Ministry of Science and Innovation (MICINN - BIO2007-60066, SAF2008-00048, SAF30542-C01-01 and SAF2010-15512), Laboratorios SALVAT, S.A., Fundación Renal Tomás de Osma, Generalitat Valenciana Prometeo 2010/005 (partially funded with ERDF), Consolider-Ingenio 2010 (MICINN - CSD2008-00005C) and by the Generalitat Valenciana through Prometeo 2014/061. Moure was funded by a predoctoral fellowship from JAE-pre CSIC. The funders provided financial support and had a role in the data collection, analysis and manuscript preparation of cellular and in vivo model of ototoxicity. The specific roles of these authors are articulated in the ‘author contributions’ section.