Research Paper Volume 11, Issue 13 pp 4382—4406

Complement component C5a induces aberrant epigenetic modifications in renal tubular epithelial cells accelerating senescence by Wnt4/βcatenin signaling after ischemia/reperfusion injury

Giuseppe Castellano 1, *, , Rossana Franzin 1, *, , Fabio Sallustio 1, 2, , Alessandra Stasi 1, , Barbara Banelli 3, , Massimo Romani 3, , Giuseppe De Palma 1, , Giuseppe Lucarelli 4, , Chiara Divella 1, , Michele Battaglia 4, , Antonio Crovace 5, , Francesco Staffieri 5, , Giuseppe Grandaliano 6, , Giovanni Stallone 6, , Pasquale Ditonno 4, , Paolo Cravedi 7, , Vincenzo Cantaluppi 8, , Loreto Gesualdo 1, ,

  • 1 Nephrology, Dialysis and Transplantation Unit, University of Bari “Aldo Moro”, Bari, Italy
  • 2 Department of Basic Medical Sciences, Neuroscience and Sense Organs, University of Bari “Aldo Moro”, Bari, Italy
  • 3 IRCCS Ospedale Policlinico San Martino, Genova, Italy
  • 4 Urology, Andrology and Renal Transplantation Unit, University of Bari, Bari, Italy
  • 5 Veterinary Surgery Unit, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy
  • 6 Nephrology, Dialysis and Transplantation Unit, Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy
  • 7 Department of Medicine, Translational Transplant Research Center, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
  • 8 Nephrology and Kidney Transplantation Unit, Department of Translational Medicine, University of Eastern Piedmont “A. Avogadro” (UPO), Novara, Italy
* Equal contribution

received: March 8, 2019 ; accepted: June 24, 2019 ; published: July 8, 2019 ;
How to Cite

Copyright: Castellano et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Epigenetic mechanisms, such as DNA methylation, affect tubular maladaptive response after Acute Kidney Injury (AKI) and accelerate renal aging. Upon ischemia/reperfusion (I/R) injury, Complement activation leads to C5a release that mediates damage; however, little is known about the effect of C5a-C5a Receptor (C5aR) interaction in Renal Tubular Epithelial Cells (RTEC).

Through a whole-genome DNA methylation analysis in cultured RTEC, we found that C5a induced aberrant methylation, particularly in regions involved in cell cycle control, DNA damage and Wnt signaling. The most represented genes were BCL9, CYP1B1 and CDK6. C5a stimulation of RTEC led to up-regulation of SA-β Gal and cell cycle arrest markers such as p53 and p21. C5a increased also IL-6, MCP-1 and CTGF gene expression, consistent with SASP development. In accordance, in a swine model of renal I/R injury, we found the increased expression of Wnt4 and βcatenin correlating with SA-β Gal, p21, p16 and IL-6 positivity. Administration of Complement Inhibitor (C1-Inh), antagonized SASP by reducing SA-β Gal, p21, p16, IL-6 and abrogating Wnt4/βcatenin activation.

Thus, C5a affects the DNA methylation of genes involved in tubular senescence. Targeting epigenetic programs and Complement may offer novels strategies to protect tubular cells from accelerated aging and to counteract progression to Chronic Kidney Disease


AhR: Aryl Hydrocarbon Receptor Signaling; AKI: Acute Kidney Injury; Bas: Basal condition; BCL9: B-Cell CLL/Lymphoma 9; C1-INH: C1-Inhibitor; CCDC6: Coiled-Coil Domain Containing 6; CKD: Chronic Kidney Disease; CTRL: Control group; CYP1B1: Cytochrome P450 Family 1 Subfamily B Member 1; DNMT: DNA methyl transferase; FDZ6: Frizzled Class Receptor 6; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; I/R: Ischemia/Reperfusion; RTEC: Renal Tubular Epithelial Cells; SASP: Senescence-Associated Secretory Phenotype; SA-βGAL: Senescence-Associated-βgalactosidase; SD: Standard Deviation; SERPINE1 (PAI-1): Serpin Family E Member 1 (Plasminogen Activator Inhibitor, Type I); TP53: Tumor Protein P53; qMSP: Quantitative Methylation Specific PCR; 5-aza: 5-aza-2’-deoxycytidine or Decitabine.