Date: Tuesday, June 14, 2016
Session Time: 4:30pm-6:00pm
Presentation Time: 4:42pm-4:54pm
Location: Ballroom A
Background: Reducing use of tissue biopsy by efficiently using non-invasive samples for continuous monitoring and evaluation of graft function would be ideal. Reports indicate continuous systemic and local molecular communication via cell free miRNA (cf-miRNA) and RNA in organisms. We aimed at identifying cf-miRNA biomarkers in plasma (systemic) and urine supernatant (local) samples for prediction of chronic renal graft dysfunction (CRAD).
Methods: Plasma (n=16) and paired urine samples (n=6) from deceased donor kidney transplant recipients (KTR) (n=16) collected 12 months post- kidney transplantation (KT) were used for the pilot study. KTRs were part of a prospective cohort with protocol biopsies and follow-up >4 years post-KT. Patients were classified as Progressors (P) vs. Non-Progressors (NP) to CRAD, based on the eGFR trend until 48- month post-KT and histological findings. Also, a set of biopsies for cause with histological diagnosis of calcineurin inhibitor toxicity (CNIT) was included for testing specificity of markers. MicroRNA PCR arrays with panel of 84 miRNA involved in fibrosis pathway were tested in the samples. Data was normalized and was analyzed using [Delta][Delta] CT method and online PCR data analysis software from SABiosciences.
Results: 18 differentially expressed plasma cf miRNA between P and NP were identified (p value < 0.05) which include miRNA like 199b-5p, 10a-5P, 29b-3p with reported functions in pro-fibrosis, ECM remodeling, and EMT, respectively. While most of these miRNA (61%) overlap with the differentially regulated miRNA in CNIT, upregulation of miR-328-3p (pvalue= 0.012; FC=3.74) was unique to CNIT group. Urine supernatants of P and NP when analyzed showed 5 differentially expressed miRNA which partially overlapped with the differentially expressed miRNA in plasma samples between these two groups. The overall expression trend of miRNA in both body fluids across the two patient groups showed both specific and shared miRNA indicative of local tissue specific expression pattern and cf miRNA mediated transfer of information across tissue.
Conclusions: Early diagnosis of CRAD is possible by detecting cf-miRNA signature in low volumes of urine and plasma samples, before graft function is affected. The identified molecular signature needs to be validated in expanded sample size.
CITATION INFORMATION: Bontha S, Gehrau R, Rhone E, King A, Gallon L, Brayman K, Maluf D, Mas V. Non-Invasive – Minimalistic Approach Using Cell-Free MicroRNAs in Paired Plasma and Urine Samples for Early Diagnosis of Chronic Renal Allograft Dysfunction. Am J Transplant. 2016;16 (suppl 3).
To cite this abstract in AMA style:Bontha S, Gehrau R, Rhone E, King A, Gallon L, Brayman K, Maluf D, Mas V. Non-Invasive – Minimalistic Approach Using Cell-Free MicroRNAs in Paired Plasma and Urine Samples for Early Diagnosis of Chronic Renal Allograft Dysfunction. [abstract]. Am J Transplant. 2016; 16 (suppl 3). https://atcmeetingabstracts.com/abstract/non-invasive-minimalistic-approach-using-cell-free-micrornas-in-paired-plasma-and-urine-samples-for-early-diagnosis-of-chronic-renal-allograft-dysfunction/. Accessed November 26, 2020.
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