March2 Alleviates Aortic Aneurysm/Dissection by Regulating PKM2 Polymerization

Circ Res. 2025 Apr 11;136(8):e73-e93. doi: 10.1161/CIRCRESAHA.124.325049. Epub 2025 Mar 13.

Abstract

Background: Aortic aneurysm/dissection (AAD) is a life-threatening disease lacking effective pharmacological treatment. Protein ubiquitination plays a pivotal role in cardiovascular diseases. However, the possible contribution of the E3 ubiquitin ligase March2 (membrane-associated RING [really interesting new gene] finger protein 2) to the cause of AAD remains elusive.

Methods: Integrated single-cell RNA sequencing analysis was conducted in human AAD tissues. Based on the screening results, we generated a mouse line of smooth muscle cell-specific March2 knockout. β-Aminopropionitrile monofumarate was used to establish AAD. Cleavage under targets and tagmentation and cleavage under targets and tagmentation-quantitative polymerase chain reaction were performed to identify possible target genes for histone H3K18 lactylation.

Results: March2 expression was downregulated in aorta from patients with AAD or β-aminopropionitrile monofumarate-induced AAD mice. β-Aminopropionitrile monofumarate-induced AAD was significantly accentuated in March2 global (March2-/-) and vascular smooth muscle cell-specific deletion (March2fl/fl; TaglnCre) mice, whereas the AAD pathology was rescued by rAAV9-SM22α (smooth muscle 22α)-March2 (recombinant adeno-associated virus serotype 9 expressing Flag-tagged March2 under SM22α promoter). March2 interacted with PKM2 (pyruvate kinase M2) to promote K33-linked polyubiquitination. Deficiency of March2 lessened PKM2 dimer-to-tetramer conversion in AAD and overtly exacerbated AAD-induced histone H3K18 lactylation in vascular smooth muscle cells by fostering glucose metabolism reprogramming, thereby promoting p53-driven apoptotic transcriptional response-a hallmark of AAD pathogenesis. TEPP-46 (tetraethyl pyrophosphate), a PKM2-specific activator, pronouncedly alleviated March2 deficiency-deteriorated AAD pathology.

Conclusions: Our findings demonstrated that March2 is a novel endogenous defender that prevents AAD by inhibiting vascular smooth muscle cell apoptosis, suggesting that March2 represents a potential therapeutic target for AAD.

Keywords: aminopropionitrile; aortic aneurysm; aortic dissection; cardiovascular diseases; ubiquitin.

MeSH terms

  • Animals
  • Aortic Aneurysm* / chemically induced
  • Aortic Aneurysm* / genetics
  • Aortic Aneurysm* / metabolism
  • Aortic Aneurysm* / pathology
  • Aortic Dissection* / chemically induced
  • Aortic Dissection* / genetics
  • Aortic Dissection* / metabolism
  • Aortic Dissection* / pathology
  • Aortic Dissection* / prevention & control
  • Carrier Proteins* / genetics
  • Carrier Proteins* / metabolism
  • Disease Models, Animal
  • Female
  • Histones / metabolism
  • Humans
  • Male
  • Membrane Proteins* / genetics
  • Membrane Proteins* / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Muscle, Smooth, Vascular / metabolism
  • Muscle, Smooth, Vascular / pathology
  • Myocytes, Smooth Muscle / metabolism
  • Myocytes, Smooth Muscle / pathology
  • Protein Multimerization
  • Pyruvate Kinase* / metabolism
  • Thyroid Hormone-Binding Proteins
  • Thyroid Hormones* / genetics
  • Thyroid Hormones* / metabolism
  • Ubiquitin-Protein Ligases* / deficiency
  • Ubiquitin-Protein Ligases* / genetics
  • Ubiquitin-Protein Ligases* / metabolism
  • Ubiquitination

Substances

  • Ubiquitin-Protein Ligases
  • Membrane Proteins
  • Thyroid Hormone-Binding Proteins
  • Thyroid Hormones
  • Carrier Proteins
  • Pkm protein, mouse
  • MG53 protein, mouse
  • Pyruvate Kinase
  • Histones