Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Effects of intracardiac delivery of aldehyde dehydrogenase 2 gene in myocardial salvage

Abstract

Intrinsic activity of aldehyde dehydrogenase (ALDH)2, a cardiac mitochondrial enzyme, is vital in detoxifying 4-hydroxy-2-nonenal (4HNE) like cellular reactive carbonyl species (RCS) and thereby conferring cardiac protection against pathological stress. It was also known that a single point mutation (E487K) in ALDH2 (prevalent in East Asians) known as ALDH2*2 reduces its activity intrinsically and was associated with increased cardiovascular diseases. We and others have shown that ALDH2 activity is reduced in several pathologies in WT animals as well. Thus, exogenous augmentation of ALDH2 activity is a good strategy to protect the myocardium from pathologies. In this study, we will test the efficacy of intracardiac injections of the ALDH2 gene in mice. We injected both wild type (WT) and ALDH2*2 knock-in mutant mice with ALDH2 constructs, AAv9-cTNT-hALDH2-HA tag-P2A-eGFP or their control constructs, AAv9-cTNT-eGFP. We found that intracardiac ALDH2 gene transfer increased myocardial levels of ALDH2 compared to GFP alone after 1 and 3 weeks. When we subjected the hearts of these mice to 30 min global ischemia and 90 min reperfusion (I-R) using the Langendorff perfusion system, we found reduced infarct size in the hearts of mice with ALDH2 gene vs GFP alone. A single time injection has shown increased myocardial ALDH2 activity for at least 3 weeks and reduced myocardial 4HNE adducts and infarct size along with increased contractile function of the hearts while subjected to I-R. Thus, ALDH2 overexpression protected the myocardium from I-R injury by reducing 4HNE protein adducts implicating increased 4HNE detoxification by ALDH2. In conclusion, intracardiac ALDH2 gene transfer is an effective strategy to protect the myocardium from pathological insults.

This is a preview of subscription content

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Fig. 1: Map of ALDH2 construct.
Fig. 2: Immunofluorescence imaging of Hoechst dye from the injection, GFP from the construct and ALDH2 from staining show effective transduction.
Fig. 3: Cardiac ALDH2 levels after 1 and 3 weeks of intracardiac transfection.
Fig. 4: Cardiac ALDH2 levels, ALDH2 activity and 4HNE protein adducts in the WT and ALDH2*2 mutant mouse hearts subjected to global ischemia-reperfusion injury after AAV9-ALDH2-GFP and AAV9-GFP transfections.
Fig. 5: Myocardial infarct size measurements of WT and ALDH2*2 mutant mouse hearts subjected to global ischemia-reperfusion injury after AAV9-ALDH2-GFP and AAV9-GFP transfections.
Fig. 6: Changes in cardiac functional indices of WT and ALDH2*2 mutant mouse hearts subjected to global ischemia-reperfusion injury after AAV9-ALDH2-GFP and AAV9-GFP transfections.

Data availability

All the data are within the manuscript. All materials were obtained from commercial vendors.

References

  1. Yoshida A, Rzhetsky A, Hsu LC, Chang C. Human aldehyde dehydrogenase gene family. Eur J Biochem. 1998;251:549–57.

    CAS  Article  Google Scholar 

  2. Raghunathan L, Hsu LC, Klisak I, Sparkes RS, Yoshida A, Mohandas T. Regional localization of the human genes for aldehyde dehydrogenase-1 and aldehyde dehydrogenase-2. Genomics. 1988;2:267–9.

    CAS  Article  Google Scholar 

  3. Braun T, Bober E, Singh S, Agarwal DP, Goedde HW. Evidence for a signal peptide at the amino-terminal end of human mitochondrial aldehyde dehydrogenase. FEBS Lett. 1987;215:233–6.

    CAS  Article  Google Scholar 

  4. Eriksson CJ, Marselos M, Koivula T. Role of cytosolic rat liver aldehyde dehydrogenase in the oxidation of acetaldehyde during ethanol metabolism in vivo. Biochem J. 1975;152:709–12.

    CAS  Article  Google Scholar 

  5. Vasiliou V, Pappa A, Petersen DR. Role of aldehyde dehydrogenases in endogenous and xenobiotic metabolism. Chem Biol Interact. 2000;129:1–19.

    CAS  Article  Google Scholar 

  6. Roede JR, Jones DP. Reactive species and mitochondrial dysfunction: mechanistic significance of 4-hydroxynonenal. Environ Mol Mutagen. 2010;51:380–90.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Esterbauer H, Schaur RJ, Zollner H. Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic Biol Med. 1991;11:81–128.

    CAS  Article  Google Scholar 

  8. Bulteau AL, Lundberg KC, Humphries KM, Sadek HA, Szweda PA, Friguet B, et al. Oxidative modification and inactivation of the proteasome during coronary occlusion/reperfusion. J Biol Chem. 2001;276:30057–63.

    CAS  Article  Google Scholar 

  9. Ferrington DA, Kapphahn RJ. Catalytic site-specific inhibition of the 20S proteasome by 4-hydroxynonenal. FEBS letters. 2004;578:217–23.

    CAS  Article  Google Scholar 

  10. Farout L, Mary J, Vinh J, Szweda LI, Friguet B. Inactivation of the proteasome by 4-hydroxy-2-nonenal is site specific and dependant on 20S proteasome subtypes. Arch Biochem Biophys. 2006;453:135–42.

    CAS  Article  Google Scholar 

  11. Akude E, Zherebitskaya E, Roy Chowdhury SK, Girling K, Fernyhough P. 4-Hydroxy-2-nonenal induces mitochondrial dysfunction and aberrant axonal outgrowth in adult sensory neurons that mimics features of diabetic neuropathy. Neurotox Res. 2010;17:28–38.

    CAS  Article  Google Scholar 

  12. Kaplan P, Tatarkova Z, Racay P, Lehotsky J, Pavlikova M, Dobrota D. Oxidative modifications of cardiac mitochondria and inhibition of cytochrome c oxidase activity by 4-hydroxynonenal. Redox Rep. 2007;12:211–8.

    CAS  Article  Google Scholar 

  13. Keller JN, Mark RJ, Bruce AJ, Blanc E, Rothstein JD, Uchida K, et al. 4-Hydroxynonenal, an aldehydic product of membrane lipid peroxidation, impairs glutamate transport and mitochondrial function in synaptosomes. Neuroscience. 1997;80:685–96.

    CAS  Article  Google Scholar 

  14. Keith RJ, Haberzettl P, Vladykovskaya E, Hill BG, Kaiserova K, Srivastava S, et al. Aldose reductase decreases endoplasmic reticulum stress in ischemic hearts. Chem Biol Interact. 2009;178:242–9.

    CAS  Article  Google Scholar 

  15. Mali VR, Ning R, Chen J, Yang XP, Xu J, Palaniyandi SS. Impairment of aldehyde dehydrogenase-2 by 4-hydroxy-2-nonenal adduct formation and cardiomyocyte hypertrophy in mice fed a high-fat diet and injected with low-dose streptozotocin. Exp Biol Med. 2014;239:610–8.

    Article  Google Scholar 

  16. Mali VR, Pan G, Deshpande M, Thandavarayan RA, Xu J, Yang XP, et al. Cardiac mitochondrial respiratory dysfunction and tissue damage in chronic hyperglycemia correlate with reduced aldehyde dehydrogenase-2 activity. PLoS One. 2016;11:e0163158.

    Article  Google Scholar 

  17. Eaton P, Li JM, Hearse DJ, Shattock MJ. Formation of 4-hydroxy-2-nonenal-modified proteins in ischemic rat heart. Am J Physiol. 1999;276:H935–43.

    CAS  PubMed  Google Scholar 

  18. Lucas DT, Szweda LI. Declines in mitochondrial respiration during cardiac reperfusion: age-dependent inactivation of alpha-ketoglutarate dehydrogenase. Proc Natl Acad Sci U S A. 1999;96:6689–93.

    CAS  Article  Google Scholar 

  19. Chen CH, Budas GR, Churchill EN, Disatnik MH, Hurley TD, Mochly-Rosen D. Activation of aldehyde dehydrogenase-2 reduces ischemic damage to the heart. Science. 2008;321:1493–5.

    CAS  Article  Google Scholar 

  20. Goedde HW, Agarwal DP, Fritze G, Meier-Tackmann D, Singh S, Beckmann G, et al. Distribution of ADH2 and ALDH2 genotypes in different populations. Hum Genet. 1992;88:344–6.

    CAS  Article  Google Scholar 

  21. Luo HR, Wu GS, Pakstis AJ, Tong L, Oota H, Kidd KK, et al. Origin and dispersal of atypical aldehyde dehydrogenase ALDH2487Lys. Gene. 2009;435:96–103.

    CAS  Article  Google Scholar 

  22. Harada S, Agarwal DP, Goedde HW, Ishikawa B. Aldehyde dehydrogenase isozyme variation and alcoholism in Japan. Pharmacol Biochem Behav. 1983;18:151–3.

    Article  Google Scholar 

  23. Crabb DW, Edenberg HJ, Bosron WF, Li TK. Genotypes for aldehyde dehydrogenase deficiency and alcohol sensitivity. The inactive ALDH2(2) allele is dominant. J Clin Invest. 1989;83:314–6.

    CAS  Article  Google Scholar 

  24. Zhong Z, Hou J, Li B, Zhang Q, Li C, Liu Z, et al. Genetic polymorphisms of the mitochondrial aldehyde dehydrogenase ALDH2 gene in a large ethnic hakka population in Southern China. Med Sci Monit. 2018;24:2038–44.

    CAS  Article  Google Scholar 

  25. Jo SA, Kim EK, Park MH, Han C, Park HY, Jang Y, et al. A Glu487Lys polymorphism in the gene for mitochondrial aldehyde dehydrogenase 2 is associated with myocardial infarction in elderly Korean men. Clin Chim Acta. 2007;382:43–7.

    CAS  Article  Google Scholar 

  26. Takagi S, Iwai N, Yamauchi R, Kojima S, Yasuno S, Baba T, et al. Aldehyde dehydrogenase 2 gene is a risk factor for myocardial infarction in Japanese men. Hypertens Res. 2002;25:677–81.

    CAS  Article  Google Scholar 

  27. Furfaro AL, Menini S, Patriarca S, Pesce C, Odetti P, Cottalasso D, et al. HNE-dependent molecular damage in diabetic nephropathy and its possible prevention by N-acetyl-cysteine and oxerutin. Biofactors. 2005;24:291–8.

    CAS  Article  Google Scholar 

  28. Zambelli VO, Gross ER, Chen CH, Gutierrez VP, Cury Y, Mochly-Rosen D. Aldehyde dehydrogenase-2 regulates nociception in rodent models of acute inflammatory pain. Sci Transl Med. 2014;6:251ra118.

    Article  Google Scholar 

  29. Pan G, Roy B, Palaniyandi SS. Diabetic aldehyde dehydrogenase 2 mutant (ALDH2*2) mice are more susceptible to cardiac ischemic-reperfusion injury due to 4-hydroxy-2-nonenal induced coronary endothelial cell damage. J Am Heart Assoc. 2021;10:e021140.

    CAS  Article  Google Scholar 

  30. Ma H, Guo R, Yu L, Zhang Y, Ren J. Aldehyde dehydrogenase 2 (ALDH2) rescues myocardial ischaemia/reperfusion injury: role of autophagy paradox and toxic aldehyde. Eur Heart J. 2011;32:1025–38.

    CAS  Article  Google Scholar 

  31. Chen CH, Ferreira JCB, Mochly-Rosen D. ALDH2 and cardiovascular disease. Adv Exp Med Biol. 2019;1193:53–67.

    CAS  Article  Google Scholar 

  32. Zincarelli C, Soltys S, Rengo G, Rabinowitz JE. Analysis of AAV serotypes 1-9 mediated gene expression and tropism in mice after systemic injection. Mol Ther. 2008;16:1073–80.

    CAS  Article  Google Scholar 

  33. Dipple KM, Crabb DW. The mitochondrial aldehyde dehydrogenase gene resides in an HTF island but is expressed in a tissue-specific manner. Biochem Biophys Res Commun. 1993;193:420–7.

    CAS  Article  Google Scholar 

  34. Anstee QM, Mantovani A, Tilg H, Targher G. Risk of cardiomyopathy and cardiac arrhythmias in patients with nonalcoholic fatty liver disease. Nat Rev Gastroenterol Hepatol. 2018;15:425–39.

    Article  Google Scholar 

  35. Bryson TD, Gu X, Khalil RM, Khan S, Zhu L, Xu J, et al. Overexpression of prostaglandin E2 EP4 receptor improves cardiac function after myocardial infarction. J Mol Cell Cardiol. 2018;118:1–12.

    CAS  Article  Google Scholar 

  36. Weber C, Neacsu I, Krautz B, Schlegel P, Sauer S, Raake P, et al. Therapeutic safety of high myocardial expression levels of the molecular inotrope S100A1 in a preclinical heart failure model. Gene Ther. 2014;21:131–8.

    CAS  Article  Google Scholar 

  37. Woitek F, Zentilin L, Hoffman NE, Powers JC, Ottiger I, Parikh S, et al. Intracoronary cytoprotective gene therapy: a study of VEGF-B167 in a pre-clinical animal model of dilated cardiomyopathy. J Am Coll Cardiol. 2015;66:139–53.

    CAS  Article  Google Scholar 

  38. DiMattia MA, Nam HJ, Van Vliet K, Mitchell M, Bennett A, Gurda BL, et al. Structural insight into the unique properties of adeno-associated virus serotype 9. J Virol. 2012;86:6947–58.

    CAS  Article  Google Scholar 

  39. Ma H, Li J, Gao F, Ren J. Aldehyde dehydrogenase 2 ameliorates acute cardiac toxicity of ethanol: role of protein phosphatase and forkhead transcription factor. J Am Coll Cardiol. 2009;54:2187–96.

    CAS  Article  Google Scholar 

  40. Gomes KM, Campos JC, Bechara LR, Queliconi B, Lima VM, Disatnik MH, et al. Aldehyde dehydrogenase 2 activation in heart failure restores mitochondrial function and improves ventricular function and remodelling. Cardiovasc Res. 2014;103:498–508.

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank Durga Dham and Saswat Saravanan for their English editing.

Funding

SSP was supported by a grant from the National Heart, Lung, and Blood Institute 1R56HL131891-01A1, 1R01HL139877-01A1, and an internal grant from Henry Ford Health System A10249.

Author information

Authors and Affiliations

Authors

Contributions

GP conducted the study, collected, analyzed, and organized the data. BR contributed to the data collection. TL and RH contributed to construct design and production and quality control. RAT contributed to manuscript editing. SSP contributed to overall project conception, experimental design, data analysis and organization and then, manuscript writing/editing and finalization. All authors reviewed the manuscript.

Corresponding author

Correspondence to Suresh Selvaraj Palaniyandi.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval

All applicable national, and/or institutional guidelines for the care and use of animals were followed.

Consent for publication

All authors have declared their consent for this publication.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Pan, G., Roy, B., Harding, P. et al. Effects of intracardiac delivery of aldehyde dehydrogenase 2 gene in myocardial salvage. Gene Ther (2022). https://doi.org/10.1038/s41434-022-00345-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1038/s41434-022-00345-2

Search

Quick links