Abramowicz, D

Beirnaert, C

Bosmans, J-L

De Schrijver, J

De Winter, B

Del Favero, J

Dendooven, Amélie

Emonds, M-P

Gielis, E.

Laukens, K

Ledeganck, K

Meysman, P

Van Laecke, Steven

Ghent University Academic Bibliography

CONCLUSIONS:Our data demonstrated that acute and chronic AMR showed clearsigns of accelerated tubular ageing (Inflammaging), with increased number of tubularp16INK4aþ senescent cells. Moreover, soluble serum factors from graft recipients suchas Complement C5amight be responsible for inducing premature tubular aging.FP694 IDENTIFICATION OF TRANSCRIPTOMIC AND MIRNOMICVARIABLES UNDERLYING THE PULMONARY FIBROSISINDUCED BY HIGH DOSES EVEROLIMUS: LOOKING FOR NEWANTI-FIBROTIC THERAPEUTIC TARGETS IN RENALTRANSPLANTATIONGianluigi Zaza3, Simona Granata2, Gloria Santoro2, Valentina Masola2,Pierluigi Carratu1, Nadia Antonucci2, Antonio Lupo21Respiratory diseases, Pulmonary Disease, university of Bari, Italy, 2Medicine, Renal Unit,University of Verona, Italy and 3Renal Unit, Medicine, University of Verona, Verona, ItalyINTRODUCTIONANDAIMS: Several studies have reported that renal transplantpatients treated with mTOR inhibitors may experience pulmonary fibrosis-associatedadverse effects (including BOOP). Although epithelial to mesenchymal transition(EMT) of airway cells has a central role in this process, the complete biological machi-nery involved is not completely clarified.METHODS: In order to improve our knowledge on this process, primary wild-typebronchial epithelial cells (BE63/3) were treated with 5 and 100 nM of EVE for 24 hours.The expression of major EMTmarkers (a-SMA, Vimentin, Fibronectin) were meas-ured by RT-PCR and western blot. Transepithelial resistance was measured by aMillicell-ERS ohmmeter. mRNA andmicroRNA profiling were performed by Illuminaand Agilent kit, respectively.RESULTS:High dosage EVE caused EMT and reduced the transepithelial resistance.Gene expression profiling showed that after treatment with EVE 100 nM, 125 geneswere differentially expressed. Enrichment analysis revealed that these genes belong to44 pathways (including collagen synthesis andmetabolism). Connective tissue growthfactor (CTGF) was a high selected gene. Treatment with 5 nM EVE did not cause anyeffect on pro-fibrotic pathways. Three miRNAs resulted up-regulated after EVE 100nM treatment and able to regulate 31 of the genes selected by the transcriptomic analy-sis (including CTGF). RT-PCR and western blot for MMP12 and CTGF validatedmicroarrays.CONCLUSIONS: These results confirmed our previous published data reporting thepro-fibrotic activity of high dosage EVE in patients and showed an EVE-related multi-factorial biological machinery associated with the above-mentioned process. CTGFseemed to have a central role in this process and a possible new therapeutic candidate.FP695 PLASMA DONOR-DERIVED CELL-FREE DNA KINETICS INSTABLE RENAL TRANSPLANT RECIPIENTS AND RECIPIENTSWITH AN ACUTE REJECTIONE Gielis7, K Ledeganck7, A Dendooven2, P Meysman6, C Beirnaert6, K Laukens6,J De Schrijver5, S Van Laecke4, M-P Emonds3, B De Winter7, J-L Bosmans1,7,J Del Favero5, D Abramowicz1,71Dept. of Nephrology and Hypertension, Antwerp University Hospital, Antwerp, Belgium,2Dept. of Pathology, Antwerp University Hospital, Antwerp, Belgium, 3Histocompatibilityand Immunogenetic Laboratory, Belgian Red Cross Flanders, Mechelen, Belgium, 4RenalDivision, Ghent University Hospital, Ghent, Belgium, 5R&D dept., Multiplicom N.V., partof Agilent Technologies, Niel, Belgium, 6Biomedical Informatics Research CenterAntwerp (Biomina), University of Antwerp, Antwerp, Belgium and 7Lab. of ExperimentalMedicine & Pediatrics, University of Antwerp, Antwerp, BelgiumINTRODUCTIONANDAIMS: After transplantation, cell-free DNA derived from thedonor organ (ddcfDNA) can be detected in the recipient’s circulation.We aimed toinvestigate plasma ddcfDNA levels in renal transplant recipients with a stable graftfunction after transplantation and transplant recipients with an acute rejection (AR)episode.METHODS: Plasma samples were collected longitudinally from day 1 until 3 monthsafter transplantation from 107 renal transplant recipients within a multicenter set-up.Renal surveillance and indication biopsies were histologically evaluated by a singlepathologist according to the Banff classification. Cell-free DNA from the donor wasquantified in plasma by next generation sequencing as a fraction of total cell-free DNAusing a targeted, multiplex PCR basedmethod for the analysis of donor-specific SNPs.For the current analysis, recipients with an AR episode and stable renal transplantrecipients without acute kidney injury on all study follow-up visits and without fluctua-tions in plasma ddcfDNA fractions were selected.RESULTS: Forty-two (39.3%) subjects were categorized as stable renal transplantrecipients. Within this group, median baseline plasma ddcfDNA levels of 0.49%(0.14%-0.96%) were reached approximately three weeks after transplantation.Fourteen (13.1%) recipients developed one or two AR episodes. Plasma ddcfDNAkinetics from 13 recipients with samples available before rejection treatment are shownin Figure 1. Four recipients developed an episode of a treated borderline rejection (Fig.1A) on surveillance or indication biopsy with median ddcfDNA fractions of 0.42%(0.31%-0.56%) at time of rejection. Six Banff IIA rejection episodes were identified infive recipients. Levels of ddcfDNA were increased during two Banff IIA episodes (Fig.1B). In one recipient, plasma ddcfDNA increased up to 1.19%while levels increased to5.17% in another recipient with a Banff IIA rejection episode already four days aftertransplantation. Six episodes of antibody-mediated rejection (ABMR) or mixed cellularAR and ABMRwere identified in four recipients (Fig. 1C). Increased ddcfDNA frac-tions were observed during four out of these six episodes with median ddcfDNA frac-tions of 20.29% (1.43%-53.64%).CONCLUSIONS:Within 3 weeks after transplantation, plasma ddcfDNA fractionsdecrease to a stable baseline level. In recipients with an AR, increased levels of ddcfDNAseem phenotype dependent as higher levels are observed during an acute ABMR or amixed cellular AR and ABMR episode compared to Borderline or Banff IIA rejectionepisodes.FP696 ISOLATED V-LESION EARLY AFTER KIDNEYTRANSPLANTATION MAY NOT TRULY REPRESENT AREJECTIONMariana Wohlfahrtova5, Petra Hruba6, Marek Novotny5, Jiri Klema2,Zdenek Krejcik7, Viktor Stranecky1, Eva Honsova3, Petra Vichova4,Ondrej Viklicky51Inst. of Inherited Metabolic Disorders, 1st Faculty of Medicine Charles University andGeneral Faculty Hospital, Prague, Czech Republic, 2Dept. of Computer Science, CzechTechnical University, Prague, Czech Republic, 3Dept. of Clinical and TransplantPathology, IKEM, Prague, Czech Republic, 4Dept. of Immunogenetics, IKEM, Prague,Czech Republic, 5Dept. of Nephrology, IKEM, Prague, Czech Republic, 6TransplantLaboratory, IKEM, Prague, Czech Republic and 7UHKT, Institute of Haematology andBlood Transfusion, Prague, Czech RepublicINTRODUCTIONANDAIMS:Acute vascular rejection (AVR) is known to be a nega-tive prognostic factor for kidney allograft survival. However, a role of early isolated v-lesion (IV) defined as intimal arteritis with minimal tubulointerstitial inflammation(TI) is unclear. While some authors believe in hidden ischemic/reperfusion injury,others are convinced of its rejection origin. To help resolve if IV truly represents acuterejection, molecular profiling of IV and T-cell mediated vascular rejection (TCMRV)was performed.METHODS:Gene expression profile of early IV (n¼6) and TCMRV (n¼4) within 1stmonth after kidney transplantation was compared usingmicroarrays (IlluminaHumanHT-12 v4 Expression BeadChips).RESULTS:Usingmicroarray-based gene expression profiling in a training set ofpatients, we identified 288 upregulated and 22 downregulated genes in early TCMRVcompared to IV (Fig.1). These dysregulated genes were primarily associated with T andB cell activation, costimulation, differentiation, receptor signalling pathway, antigenprocessing and presentation indicating higher innate and adaptive immune responseand inflammation in early TCMRV biopsies. Hierarchical clustering (Fig 2) and princi-pal component analysis applied to the whole microarray data indicated a clear distinc-tion in kidney allografts with TCMRV and IV. Our findings were verified in anindependent validation set of patients (n¼20) using RT-qPCR and confirmedincreased expression of genes associated with immunity and inflammation in TCMRV.CONCLUSIONS: Early IV has transcriptional profile of immune injury of significantlylower extend compared toTCMRV. According to this genome-wide transcriptomeanalysis early IVmay feature non-rejection phenotype. Our findings provide a resourcefor better understanding the origin of IV and are in agreement with recent literatureevidence which shows that intimal arteritis has probably less importance in diagnosticsof TCMR than TI infiltrate. The study results call for reassessment of current histopa-thology classification which considers intimal arteritis to be at least grade 2 of TCMRirrespective of TI inflammation. Supported byMinistry of Health of the CzechRepublic, grant nr. 15-26519A and 17-29992A.Abstracts Nephrology Dialysis Transplantationi280 | AbstractsDownloaded from https://academic.oup.com/ndt/article-abstract/33/suppl_1/i280/4997798by Ghent University useron 20 July 2018

Plasma donor-derived cell-free DNA kinetics in stable renal transplant recipients and recipients with an acute rejection

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Plasma donor-derived cell-free DNA kinetics in stable renal transplant recipients and recipients with an acute rejection

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