ASN's Mission

To create a world without kidney diseases, the ASN Alliance for Kidney Health elevates care by educating and informing, driving breakthroughs and innovation, and advocating for policies that create transformative changes in kidney medicine throughout the world.

learn more

Contact ASN

1401 H St, NW, Ste 900, Washington, DC 20005

email@asn-online.org

202-640-4660

The Latest on Twitter

Kidney Week

Abstract: FR-PO139

Higher Post-COVID-19 Urinary Mitochondrial DNA Level: Serves a Biomarker of Mitochondrial Distress and Inducer of Inflammatory Cytokines Secretion in Peripheral Blood Mononuclear Cells (PBMCs)

Session Information

  • AKI: Mechanisms - II
    November 04, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
    Abstract Time: 10:00 AM - 12:00 PM

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Prasad, Narayan, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, Uttar Pradesh, India
  • Yadav, Brijesh, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, Uttar Pradesh, India
Background

Severe acute respiratory corona virus-2(SARS-CoV-2) affected multiple organs including Kidney. SARS-CoV-2 open reading frame protein 3a induces necroptosis in infected cell leading release of mt-DNA, which binds to TLR9 and trigger innate immunity, which may lead to acute allograft injury.

Methods

Sixty-six live related renal allograft recipient previously hospitalized with SARS-CoV-2 infection were recruited after 2-3week of discharge. Patients were categorized either in non-AKI(n=47)orAKI(n=19) group, if hospitalization serum creatinine level was >30% of preCovid serum creatinine. A 50ml urine sample was collected for the umt-DNA gene NADH-ubiquinone oxidoreductase chain1(ND-1) and nuclear 36B4 gene quantification by RT-PCR and urine N-GAL measurement by ELISA. A 10ml blood sample from 10healthy volunteers was collected for PBMCs isolation. A 1x106 PBMCs were stimulated for 24hrs. with 1µg/ml of urinary DNA or CpG oligodeoxynucleotide(5’-tcgtcgttttcggcgc:gcgccg-3’) in duplicate.Unstimulated PBMCs served as control. The gene expression of IL-10,IL-6,MYD88 was analyzed by the RT-PCR and IL-6,IL-10 level in supernatants by the ELISA.

Results

The precovid creatinine in non-AKI vs AKI patient was 1.06±0.20vs0.97±0.27, p=.14; at hospitalization 1.27±0.18vs1.84 ±0.37,p<.001; at discharge 1.09±0.20vs1.11±0.32mg/dl, p=0.73. The mean ND-1 gene Ct in non-AKI vs AKI was 21.77±3.60vs19.44±2.58a.u, p=.013. The normalized ND-1 Ct in non-AKI vs AKI was 0.89+0.14vs0.79±0.11a.u, P=0.007. The median urinary N-GAL level in non-AKI vs AKI group was 212.78 (range,219.8-383.06) vs 453.5 (range,320.2-725.02; p=0.015)ng/ml. The area under curve of ND-1 Ct gene was 0.73, normalized ND-1 Ct was 0.71, uNGAL was 0.66 and normalized uNGAL was 0.68 for detecting the AKI. The IL-10 gene expression was downregulated in umt-DNA treated PBMCs compared to control (-3.5±0.40vs1.02±0.02, p<0.001).IL-6 and Myd88 gene expression was upregulated. The culture supernatant IL-10 and IL-6 level in umt-DNA treatment PBMCs vs control was 10.65±2.02 vs 30.3±5.47, p=0.001; and 200.2±33.67 vs 47.6±12.83, p=0.001 respectively.

Conclusion

Quantification of umt-DNA can detect the post covid19 mitochondrial distress with higher sensitivity compare to uNGAL. umt-DNA induces robust inflammatory response in PBMCs may exacerbate the post-Covid19 allograft injury.

Funding

  • Government Support – Non-U.S.