PDK4 Deficiency Induces Intrinsic Apoptosis in Response to Starvation in Fibroblasts from Doberman Pinschers with Dilated Cardiomyopathy

Kathryn Taggart, Amara Estrada, Patrick Thompson, Francisco Lourenco, Sara Kirmani, Silveli Suzuki-Hatano, Christina A. Pacak

Research output: Contribution to journalArticlepeer-review

Abstract

The Doberman pinscher (DP) canine breed displays a high incidence of idiopathic, nonischemic dilated cardiomyopathy (DCM) with increased mortality. A common mutation in DPs is a splice site deletion in the pyruvate dehydrogenase kinase 4 (PDK4) gene that shows a positive correlation with DCM development. PDK4, a vital mitochondrial protein, controls the switch between glycolysis and oxidative phosphorylation based upon nutrient availability. It is likely, although unproven, that DPs with the PDK4 mutation are unable to switch to oxidative phosphorylation during periods of low nutrient availability, and thus are highly susceptible to mitochondrial-mediated apoptosis. This study investigated cell viability, mitochondrial stress, and activation of the intrinsic (mitochondrial mediated) apoptotic pathway in dermal fibroblasts from DPs that were healthy (PDK4wt/wt), heterozygous (PDK4wt/del), and homozygous (PDK4del/del) for the PDK4 mutation under conditions of high (unstarved) and low (starved) nutrient availability in vitro. As hypothesized, PDK4wt/del and PDK4del/del cells showed evidence of mitochondrial stress and activation of the intrinsic apoptotic pathway following starvation, while the PDK4wt/wt cells remained healthy and viable under these conditions. Adeno-associated virus (AAV) PDK4-mediated gene replacement experiments confirmed cause-effect relationships between PDK4 deficiency and apoptosis activation. The restoration of function observed following administration of AAV-PDK4 provides strong support for the translation of this gene therapy approach into the clinical realm for PDK4-affected Dobermans.

Original languageEnglish (US)
Pages (from-to)182-191
Number of pages10
JournalBioResearch Open Access
Volume6
Issue number1
DOIs
StatePublished - Dec 2017
Externally publishedYes

Bibliographical note

Funding Information:
The authors thank The Doberman Pinscher Club of America, Cooper’s Fund, University of Florida Vector Core, and Dr. Arun Srivastava for kindly sharing the AAV2 quad-mutant capsid; and the Boehringer Ingelheim Veterinary Scholars Fund supported this study.

Publisher Copyright:
© 2017, Mary Ann Liebert, Inc.

Keywords

  • AAV
  • cardiomyopathy
  • dilated Doberman Pinschers
  • mitochondria
  • PDK4
  • pyruvate dehydrogenasekinase

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