Mitochondrial DNA: A Biomarker of Disease Severity in Necrotizing Enterocolitis

Edoardo Bindi; Bo Li; Haitao Zhou; Maarten Janssen Lok; Mashriq Alganabi; Rossella Angotti; Agostino Pierro

doi:10.1055/s-0039-1697910

European Journal of Pediatric Surgery, Table of Contents

Eur J Pediatr Surg 2020; 30(01): 085-089
DOI: 10.1055/s-0039-1697910

Original Article

Georg Thieme Verlag KG Stuttgart · New York

Mitochondrial DNA: A Biomarker of Disease Severity in Necrotizing Enterocolitis

Edoardo Bindi

¹Division of General and Thoracic Surgery, Translational Medicine Program, The Hospital for Sick Children, Toronto, Ontario, Canada

²Division of Pediatric Surgery, Department of Medical Sciences, Surgical Sciences and Neurosciences, Hospital of “Santa Maria alle Scotte,” Siena, Italy

,

Bo Li

¹Division of General and Thoracic Surgery, Translational Medicine Program, The Hospital for Sick Children, Toronto, Ontario, Canada

,

Haitao Zhou

¹Division of General and Thoracic Surgery, Translational Medicine Program, The Hospital for Sick Children, Toronto, Ontario, Canada

,

Maarten Janssen Lok

¹Division of General and Thoracic Surgery, Translational Medicine Program, The Hospital for Sick Children, Toronto, Ontario, Canada

,

Mashriq Alganabi

¹Division of General and Thoracic Surgery, Translational Medicine Program, The Hospital for Sick Children, Toronto, Ontario, Canada

,

Rossella Angotti

²Division of Pediatric Surgery, Department of Medical Sciences, Surgical Sciences and Neurosciences, Hospital of “Santa Maria alle Scotte,” Siena, Italy

,

Agostino Pierro

¹Division of General and Thoracic Surgery, Translational Medicine Program, The Hospital for Sick Children, Toronto, Ontario, Canada

› Author Affiliations

Abstract

Introduction There is a need to develop sensitive markers to diagnose or monitor the severity of intestinal damage in necrotizing enterocolitis (NEC). Mitochondrial deoxyribonucleic acid (mtDNA) is increased in the intestine and blood of adults in response to intestinal ischemia and can trigger secondary organ damage. We hypothesize that mtDNA is increased during experimental NEC and that mtDNA levels are correlated to the degree of intestinal injury.

Materials and Methods NEC was induced in C57BL/6 mice (n = 18) (approval: 44032) by gavage feeding with hyperosmolar formula, hypoxia, and lipopolysaccharide administration from postnatal day (P) 5 to 9. Breastfed pups served as control (n = 15). Blood was collected by cardiac puncture and terminal ileum was harvested on P9. Reverse transcription quatitative polymerase chain reaction was used to measure mtDNA (markers COX3, CYTB, ND1) and inflammatory cytokines (interleukin 6 [IL-6] and tumor necrosis factor-α[TNF-α]) in blood and ileum. Intestinal injury was scored blindly by four investigators and classified as no/minor injury (score 0 or 1) or NEC (score ≥2).

Results mtDNA is significantly increased in gut and blood of NEC mice (p < 0.05). Furthermore, mtDNA increases in intestine and blood proportionally to the degree of intestinal injury as indicated by a positive correlation with histological scoring and inflammation (r = 0.6; p < 0.05) (expression of IL-6 and TNF-α).

Conclusion Following NEC intestinal injury, mtDNA is released from the intestine into circulation. The blood level of mtDNA is related to the degree of intestinal injury. mtDNA can be a novel marker of intestinal injury and can be useful for monitoring the progression of NEC.

Keywords

necrotizing enterocolitis - mitochondrial DNA - disease severity - biomarker

Full Text

References

References
1 Horbar JD, Edwards EM, Greenberg LT. , et al. Variation in performance of neonatal intensive care units in the United States. JAMA Pediatr 2017; 171 (03) e164396
2 Goldstein GP, Sylvester KG. Biomarker discovery and utility in necrotizing enterocolitis. Clin Perinatol 2019; 46 (01) 1-17
3 Hauser CJ, Sursal T, Rodriguez EK, Appleton PT, Zhang Q, Itagaki K. Mitochondrial damage associated molecular patterns from femoral reamings activate neutrophils through formyl peptide receptors and P44/42 MAP kinase. J Orthop Trauma 2010; 24 (09) 534-538
4 Zhang Q, Itagaki K, Hauser CJ. Mitochondrial DNA is released by shock and activates neutrophils via p38 map kinase. Shock 2010; 34 (01) 55-59
5 Hu Q, Ren H, Ren J. , et al. Released mitochondrial DNA following intestinal ischemia reperfusion induces the inflammatory response and gut barrier dysfunction. Sci Rep 2018; 8 (01) 7350
6 Gu X, Yao Y, Wu G, Lv T, Luo L, Song Y. The plasma mitochondrial DNA is an independent predictor for post-traumatic systemic inflammatory response syndrome. PLoS One 2013; 8 (08) e72834
7 Barlow B, Santulli TV, Heird WC, Pitt J, Blanc WA, Schullinger JN. An experimental study of acute neonatal enterocolitis--the importance of breast milk. J Pediatr Surg 1974; 9 (05) 587-595
8 Zani A, Cordischi L, Cananzi M. , et al. Assessment of a neonatal rat model of necrotizing enterocolitis. Eur J Pediatr Surg 2008; 18 (06) 423-426
9 Miyake H, Li B, Lee C. , et al. Liver damage, proliferation, and progenitor cell markers in experimental necrotizing enterocolitis. J Pediatr Surg 2018; 53 (05) 909-913
10 Sylvester KG, Ling XB, Liu GY. , et al. A novel urine peptide biomarker-based algorithm for the prognosis of necrotising enterocolitis in human infants. Gut 2014; 63 (08) 1284-1292
11 Ji J, Ling XB, Zhao Y. , et al. A data-driven algorithm integrating clinical and laboratory features for the diagnosis and prognosis of necrotizing enterocolitis. PLoS One 2014; 9 (02) e89860
12 Gephart SM, Gordon PV, Penn AH. , et al. Changing the paradigm of defining, detecting, and diagnosing NEC: perspectives on Bell's stages and biomarkers for NEC. Semin Pediatr Surg 2018; 27 (01) 3-10
13 Evennett NJ, Hall NJ, Pierro A, Eaton S. Urinary intestinal fatty acid-binding protein concentration predicts extent of disease in necrotizing enterocolitis. J Pediatr Surg 2010; 45 (04) 735-740
14 Pergialiotis V, Konstantopoulos P, Karampetsou N. , et al. Calprotectin levels in necrotizing enterocolitis: a systematic review of the literature. Inflamm Res 2016; 65 (11) 847-852
15 Skulachev VP. Mitochondrial physiology and pathology; concepts of programmed death of organelles, cells and organisms. Mol Aspects Med 1999; 20 (03) 139-184
16 Thurairajah K, Briggs GD, Balogh ZJ. The source of cell-free mitochondrial DNA in trauma and potential therapeutic strategies. Eur J Trauma Emerg Surg 2018; 44 (03) 325-334
17 Gu X, Wu G, Yao Y. , et al. Intratracheal administration of mitochondrial DNA directly provokes lung inflammation through the TLR9-p38 MAPK pathway. Free Radic Biol Med 2015; 83: 149-158
18 Xie L, Liu S, Cheng J, Wang L, Liu J, Gong J. Exogenous administration of mitochondrial DNA promotes ischemia reperfusion injury via TLR9-p38 MAPK pathway. Regul Toxicol Pharmacol 2017; 89: 148-154
19 Zhang J, Chen X, Liu Z. , et al. Association between plasma mitochondrial DNA and sterile systemic inflammatory response syndrome in patients with acute blunt traumatic injury. Int J Clin Exp Med 2017; 10 (02) 3254-3262
20 Alganabi M, Lee C, Bindi E, Li B, Pierro A. Recent advances in understanding necrotizing enterocolitis. F1000 Res 2019; 8: 8