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Molecular Phenotypes of Acute Kidney Injury in Kidney Transplants

† †‡ † † Konrad S. Famulski,* Declan G. de Freitas, Chatchai Kreepala, Jessica Chang, † † † † Joana Sellares, Banu Sis,* Gunilla Einecke,§ Michael Mengel,* Jeff Reeve,* and †| Philip F. Halloran

Departments of *Laboratory Medicine and Pathology and |Medicine, University of Alberta, Edmonton, Alberta, Canada; †Alberta Transplant Applied Genomic Centre, Edmonton, Alberta, Canada; ‡Manchester Royal Infirmary, Manchester, United Kingdom; and §Hannover Medical School, Hannover, Germany

ABSTRACT Little is known regarding the molecular phenotype of kidneys with AKI because biopsies are performed infrequently. However, all kidney transplants experience acute injury, making early kidney transplants an excellent model of acute injury, provided the absence of rejection, because donor kidneys should not have CKD, post-transplant biopsies occur relatively frequently, and follow-up is excellent typically. Here, we used histopathology and microarrays to compare indication biopsies from 26 transplants with acute injury with 11 pristine protocol biopsies of stable transplants. Kidneys with acute injury showed increased ex- pression of 394 transcripts associated with the repair response to injury, including many -like injury molecules tissue, remodeling molecules, and inflammation molecules. Many other also pre- dicted the phenotype, including the acute injury biomarkers HAVCR1 and IL18. Pathway analysis of the injury-repair transcripts revealed similarities to cancer, development, and cell movement. The injury-repair transcript score in kidneys with acute injury correlated with reduced graft function, future renal recovery, brain death, and need for dialysis, but not with future graft loss. In contrast, histologic features of acute tubular injury did not correlate with function or with the molecular changes. Thus, the transcripts associ- ated with repair of injury suggest a massive coordinated response of the kidney parenchyma to acute injury, providing both an objective measure for assessing the severity of injury in kidney biopsies and validation for many biomarkers of AKI.

J Am Soc Nephrol 23: ccc–ccc, 2012. doi: 10.1681/ASN.2011090887

Although AKI is an important problem,1–3 native This prospective study of kidney transplants with kidneys with AKI are rarely biopsied, limiting studies AKI was undertaken to define the transcripts induced of AKI biomarkers to urine and body fluids.4 How- by human AKI and their clinical correlations. We ever, kidney transplants offer a unique opportunity previously analyzed the correlates of estimated GFR to study the injury-repair response to AKI because (eGFR) in transplant biopsies, but those were mainly all transplants experience AKI, CKD is excluded, and numerous indication biopsies are performed to exclude rejection, guided by the international Received September 2, 2011. Accepted January 10, 2012.

Banff histopathology consensus system. Moreover, Published online ahead of print. Publication date available at transplants have detailed function assessments and www.jasn.org. fi are followed inde nitely. AKI induced by transplan- Present address: Joana Sellares, Hospital de la Vall d' Hebron, 5–7 tation can be modeled in rodent kidney isografts, Barcelona, Spain. as recently reviewed.8 The mouse studies showed fl Correspondence: Dr. Philip F. Halloran, Alberta Transplant Ap- that some transcript changes in transplants re ect plied Genomics Centre, University of Alberta, 250 Heritage AKI, but other transcripts reflect the acute phase Medical Research Centre, Edmonton, Alberta T6G 2S2. Email: response to the surgical procedure.5 The picture is [email protected] further complicated in aging kidneys.9,10 Copyright © 2012 by the American Society of Nephrology

J Am Soc Nephrol 23: ccc–ccc, 2012 ISSN : 1046-6673/2305-ccc 1 CLINICAL RESEARCH www.jasn.org late biopsies with atrophy-fibrosis, not AKI.11 We hypothesized AKI cohort versus those with T cell–mediated rejection that indication biopsies from early kidney transplants, selected to (TCMR), borderline rejection, other diseases and pristine pro- exclude kidneys with rejection and diseases, will reveal the molec- tocol biopsies (Table 2). Because of the selection criteria, the ular features of AKI. Our strategy was to neutralize the ubiquitous pure AKI biopsies and pristine protocol biopsies had little in- minor changes inevitably induced by the transplant process by flammation or atrophy fibrosis, and the pristine protocol bi- comparing AKI kidneys to histologically pristine protocol biopsies opsies had no acute tubular injury features. of stable transplants. This study aimed to elucidate the molecular Three of 35 grafts with TCMR failed, but all had later changes that reflect clinically significant functional disturbance biopsies showing ABMR or mixed rejection. The 34 other and their relationship with histopathology changes and prognosis. kidney failures after indication biopsies reflected diseases that emerged late after transplantation such as ABMR or recurrent GN.15,16 RESULTS The mean eGFR at the time of biopsy was low in the AKI cohort (mean 26) and in TCMR and borderline rejection Selection of Biopsies from Kidney Transplants biopsies, compared with other indication biopsies (Table 2). with Pure AKI However, by 6 months after biopsy, these kidneys showed the We identified AKI kidneys from a prospective study of 234 most improvement (DeGFR). As expected, the pristine protocol kidney transplant biopsies for clinical indications from 173 biopsy cohort had higher mean eGFR at the time of biopsy than consenting patients. We excluded kidneys with rejection and the AKI cohort, and showed little improvement over time. kidney disease by histologic criteria, and also excluded those Kidneys with pure AKI in the early post-transplant period with nondiagnostic suspicious histologic lesions. This identified a manifest impaired function at biopsy but good recovery and “pure AKI” cohort of 28 biopsies from 26 patients, with mean prognosis, and are suitable for defining the injury-repair response eGFR at biopsy of 26 ml/min (range, 8–52) (Table 1). Of 28 to clinically significant AKI. biopsies, 21 displayed features that pathologists believe reflect acute tubular injury.12,13 Fifteen were from deceased donor kid- Defining the Injury-Repair Response Associated neys, of which eight manifested delayed graft function (DGF) Transcripts in the Pure AKI Biopsies (i.e., initial dialysis dependency). The median follow-up after bi- We compared microarray results from AKI biopsies versus opsy was 3.9 years. The extent of histologic tubule injury changes pristine protocol biopsies to define the features of clinically can be assessed for its correlation with molecular changes and significant injury. This comparison identified 946 probe sets, function because these kidneys were not chosen on the basis of including 394 nonredundant genes whose expression was histologic criteria for tubular injury; rather, they were chosed increased at least $1.5-fold higher in AKI than in pristine only by time post-transplant and exclusion of diseases. protocol biopsies (Supplemental Table 1), at the false discov- During follow-up, only one kidney from the AKI cohort ery rate (FDR) of 0.05. We named these injury-repair response– progressed to end stage kidney failure at 30 months, attributed associated transcripts (IRRATs). to antibody-mediated rejection (ABMR) caused by nonadher- For detailed study, we selected from the 394 IRRATs the 30 ence, and one patient died with a functioning graft. with the highest fold increase compared with the pristine Avalidation set was generated to test the molecular features protocol biopsies (Table 3). The expression of the individual (Table 1). The results below refer exclusively to the original top 30 IRRATs in AKI biopsies was 2.7- to 7.9-fold higher than AKI cohort, and the validation set is analyzed at the end of the in pristine protocol biopsies. Four of the top 30 IRRATs Results section. (LCN2, AKAP12, FOS, and EGR1) had previously been iden- tified as biomarkers of AKI, on the basis of detection of the Selection of Controls product in urine or blood,17–19 or mRNA expression in hu- Our strategy was to distinguish clinically significant AKI from the man kidney.20,21 The complete set of 394 IRRATs included minor injury that accompanies every transplant. Thus, controls four other known AKI biomarkers (CLU, CXCL1, CYR61, and consisted of protocol kidney transplant biopsies selected from a THSB1) but did not include some known biomarkers such as previously published cohort14 with clinically appropriate stable HAVCR1 and IL18. function at 6 weeks post-transplant, with no indications for bi- Although the IRRATs were defined without reference to opsy. After excluding biopsies with any histologic abnormalities, GFR measurements, they correlated strongly with eGFR. Thus, including acute tubular injury changes (Table 1), we were left expression of 26 of the top 30 individual IRRATs correlated with a set of 11 pristine protocol biopsies similar in age to the with reduced eGFR at biopsy, and 24 of 30 also correlated with AKI kidneys, including both living and deceased donor kidneys. eGFR recovery 6 months after biopsy (Table 3). After excluding the top 30 IRRATs, there were 138 additional IRRATs whose Histopathology, Function, and Survival expression correlated with eGFR, with correlation coefficients of Kidneys with AKI ranging from 20.82 to 20.50 (Table 4). Histology scores for inflammation (i-Banff, i-total) and Mouse orthologs of the 394 IRRATs had extensive overlap atrophy fibrosis (ci, ct) were compared for biopsies from the with transcripts previously shown to be increased in mouse AKI,

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Table 1. Demographics of patients selected for early AKI from biopsies for clinical indications and stable early transplants selected from protocol biopsies P Value (Pure P Value (Validation Pure AKI Pristine Protocol Validation AKI versus Set versus Demographics Cohort Biopsy Cohort Set Pristine Protocol Pristine Protocol Biopsies) Biopsies) Patient characteristics at diagnosis number of patients 26 11 27 mean recipient age (yr) 52 (16–75) 49 (29–70) 49 (21–68) .0.05 .0.05 primary disease GN 9 (32) 2 (18.2) 3 (11) .0.05 .0.05 diabetic nephropathy 4 (14) 1 (9.1) 10 (37) .0.05 .0.05 others 9 (32) 4 (36.4) 14 (52) .0.05 .0.05 unknown etiology 6 (21) 4 (36.4) 13 (48) .0.05 .0.05 mean donor age (yr) 50 (22–69) 53 (20–82) 43 (16–59) .0.05 .0.05 mean donor age (living donor) 46 (37–56) 44 (35–52) 44 (16–59) .0.05 .0.05 mean donor age (deceased donor) 53 (22–69) 55 (20–82) 42 (26–56) .0.05 .0.05 donor type deceased/living (% deceased 15/ 11 (57.6) 9/2 (81.8) 8/19 (30) .0.05 0.004 donor transplants) delayed graft function rate 8/26(31) 2/11 (18) 5/27 (19) .0.05 .0.05 median time from transplant to biopsy (d) 16 (6–42) 42 24 (7–42) ,0.001 ,0.001 median duration follow-up postbiopsy (d) 1414 (98–2155) 2000 (732–2932) 727 (1–2146) 0.006 ,0.001 allograft failure 2/26 (7.7)a 1/11 (9.1)b 2/27 (7.4)c .0.05 .0.05 Clinical characteristics at time of biopsy number of biopsies 28 11 28 indication for biopsy as indicated by clinician concerns about renal function 20 (71) (protocol) 25 (89) other or unknown 8 (29) (protocol) 3 (11) histologic diagnosis acute tubular injury ($5% acute tubular 21 (75) 0 (0) 18 (64) ,0.001 ,0.001 injury score) no major abnormalities 7 (25) 11 (100) 10 (36) ,0.001 ,0.001 mean eGFRd at the time of biopsy 26 (8–52) 49 (23–89) 31 (10–46) ,0.001 ,0.001 immunosuppressive drug regimen MMF, tacrolimus, steroids 11 (39.3) 0 (0) 5 (18) 0.017 .0.05 MMF, tacrolimus 8 (28.6) 0 (0) 7 (25) .0.05 .0.05 MMF, cyclosporine, steroids 3 (10.7) 8 (72.7) 7 (25) ,0.001 0.010 other 6 (21.4) 3 (27.3) 9 (32) .0.05 .0.05 Data are shown as n (%) or median (range) unless otherwise indicated. MMF, mycophenolate mofetil. aThe cause of failure: death with functioning graft, and ABMR in a nonadherent patient 30 months post-transplant. bThe cause of failure: death with functioning graft. cThe cause of failure: death with functioning graft, and membranoproliferative GN at 60 months post-transplant. deGFR obtained using the four-variable MDRD formula (in milliliters per minute per 1.73 m2). either isografts (n=124, P,0.001) or native kidneys with ischemic (see top of Figure 1). Nine of 10 kidneys with DGF had IRRAT ATN (n=254, P,0.0001).5 Of 18 mouse orthologs of the top 30 scores in the top tertile (P,0.05). The top tertile also con- human IRRATs, 17 were increased in mouse AKI models5 (Table tained nine biopsies from deceased donors (P,0.05). Kidneys 5). Six of those belonged to the previously published injury-repair from deceased donors had higher IRRAT scores than kidneys inducedsetsIRIT(I)andIRIT(L).5 Thus, IRRAT changes in AKI from living donors (3.0 versus 1.6, P,0.05). are largely conserved between human and mouse. Whereas high IRRATscores were associated with low eGFR, acute tubular injury features were not; they were present in 6 of Relationship of the IRRAT Score to Histology and 9 in the lowest tertile, 6 of 9 in the middle tertile, and 9 of 10 in Function in Injured Transplants the upper tertile (not significant). Histologic features of acute The 28 AKI biopsies were arranged by increasing IRRATscore tubular injury were also not associated with DGF or donor (geometric mean of the fold increase in the top 30 IRRATs type. However, inflammation in AKI biopsies, albeit mild by versus control nephrectomies) to compare the IRRAT scores definition, was nevertheless highest in the top tertile: the mean with DGF, eGFR, and histopathology features of acute tubular inflammation scores (i-total) were 2% for the lowest tertile, 5% injury (Figure 1). We discretized the IRRATscores into tertiles for the middle tertile, and 13% for the top tertile (P,0.05).

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Table 2. Histology and kidney function in 28 AKI kidney biopsies compared with 206 other transplant biopsies for clinical indications Pure AKI TCMR Borderline Remaining Pristine Protocol Feature (n=28) (n=35) (n=23) Biopsies (n=148)a Biopsies (n=11) Histologic lesions (continuous scores, % of cortex) inflammation (i-Banff) 4 26b 8.9 7.9 3.9 inflammation (i-total) 7 45b 28b 23b 3.9 scarring (ci) 7 24c 27b 22b 0b atrophy (ct) 6 23b 26b 22b 0.5c Grafts failed during follow-up 13 1 34 0 (excluding death with function)d Median follow-up time after first biopsy (yr) 3.9 4.0 4.2 3.8 6.3 Renal function (eGFR, ml/min) eGFR at biopsy 26 33 34 35c 49c eGFR at 6 mo 51e 42e 40 38c 55 DeGFR by 6 mo 25 9f 5b 2b 6b AKI biopsies were compared with the remaining biopsy groups. Kruskal–Wallis test with Dunn’s correction for multiple comparisons. aRemaining biopsies included diagnoses of ABMR, mixed ABMR/TCMR, GN, polyoma virus, and other nonrejecting biopsies. bP,0.001. cP,0.01. dFailed grafts are represented by one biopsy per patient. eIndicates significant difference between eGFR at biopsy and 6-month eGFR, by the paired t test. fP,0.05.

The IRRAT score in the AKI biopsies anticorrelated with (r=0.9 and r=0.7, P,0.0001, respectively). UMOD, CTS3, and the eGFR at biopsy (20.77, P,0.001) and correlated with FABP1 had no positive correlation with the IRRAT score. the increase in eGFR by 6 months (0.62, P,0.001). The acute We re-examined all probe sets passing the interquartile tubular injury score by histology did not correlate with eGFR range filter (n=12,552) to identify transcripts missed by the or the increase in GFR (Figure 2). initial filtering rules, comparing the binary phenotype of AKI In AKI kidneys, the IRRAT score is associated with DGF, versus pristine protocol biopsies. Our goal was to identify all brain death, impairment in eGFR, future recovery of function, transcripts that differ between these phenotypes by their P value and interstitial inflammation. The histologic features that and by their ability to predict these phenotypes. In Figure 3, the pathologists believe represent acute tubular injury were not x-axis gives the uncorrected P values for the comparison of phe- related to DGF,deceased donation, eGFR, recovery of function, notypes, whereas the y-axis gives the area under the curve for or IRRAT scores. prediction (AUC). The transcripts positively associated with the phenotype have an AUC .0.5. If negatively associated, the AUC Cellular Expression and Functional Annotation is ,0.5. An AUC at or near 0.5 signifies no discriminatory power. of IRRATs Apart from the top 30 IRRATs (shown as yellow dots), other Functional annotation of the top 30 IRRATs using Ingenuity redundant transcripts (n=1424) had high predictive values for Pathway Analysis (Table 6) indicated that the pathways over- the AKI (AUC $0.7). HAVCR1 and IL18 (pink dots) now represented included cancer; cellular movement, including showed correlations similar to many top 30 IRRATs. After migration of tumor cells; tissue development, including cell adjustment for the FDR, 930 transcripts remained, including adhesion; and acute phase response. The complete list of 394 all IRRATs. Another 1785 transcripts (after the FDR adjust- nonredundant IRRATs yielded similar results. Most IRRATs ment) were highly negatively associated with the AKI pheno- are expressed in parenchymal cells of kidney or in a broad ex- type (AUC #0.3). pression in many cells, making kidney parenchymal expression As an additional control, we examined these relationships likely. Relatively few were expressed in inflammatory cells. after random shuffling of the phenotype labels of AKI and protocol biopsies. No transcript was significant at FDR=0.05 Many Other Transcripts Correlate with IRRATs and when the labels were randomly shuffled. Predict AKI Using our statistical filtering rules, some reported biomarkers Relationship between IRRATs, Histology, and Function of AKI were not found in the top 30 or the complete 394 IRRAT in the Validation Set lists, including HAVCR1 (KIM1), IL18, UMOD, CTS3, or For validation of the molecular studies, we identified a second, FABP1. HAVCR1 did not meet the fold cutoff criteria, and IL18 independent set of kidneys with AKI, selected only as being the did not pass the statistical filter. Nevertheless, HAVCR1 and IL18 next consecutive kidney biopsies meeting the same criteria in expression correlated with the IRRAT score in the AKI biopsies this ongoing study (Table 1). The validation set of 27 kidneys

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Table 3. Top 30 IRRATs ordered by fold change in AKI biopsies versus pristine protocol biopsies Fold Change Raw Signal Correlation Correlation P Gene Name AKI versus in Protocol with eGFR at with DGFR at Symbol Value Protocol Biopsies Biopsies Biopsy in AKI 6 Mo in AKI ITGB6 b6 7.9 0.001 72 20.69 0.54 SERPINA3 Serine (or cysteine) proteinase inhibitor, clade A 6.1 0.001 252 20.71 0.47 MTND6 NADH dehydrogenase, subunit 6 (complex I) 5.6 0.003 67 20.51 0.43 OLFM4 Olfactomedin 4 5.5 0.001 53 20.74 0.46 PTX3 Pentaxin-related gene, rapidly induced by IL-1b 5.3 0.002 15 20.58 0.52 LCN2a lipocalin 2 (oncogene 24p3) 4.3 0.004 89 20.57 0.50 VCAN 2 () 4.1 0.004 47 20.35b 0.38 LTF Lactotransferrin 4.1 0.032 441 20.63 0.54 SLPI Secretory leukocyte protease inhibitor 4.1 0.013 334 20.70 0.52 (antileukoproteinase) ADAMTS1 A disintegrin-like and metalloprotease 4.0 0.000 110 20.64 0.52 (reprolysin type) with thrombospondin type 1 motif, 1 MEGF11 Multiple EGF-like domains 3.8 0.001 12 20.68 0.59 CDH6 6, type 2, K-cadherin (fetal kidney) 3.7 0.000 105 20.72 0.56 PI15 Peptidase inhibitor 15 3.6 0.008 17 20.40 0.23b FCGR3A Fc fragment of IgG, low affinity IIIa, receptor for (CD16) 3.4 0.002 41 20.30b 0.30b NNMT Nicotinamide N-methyltransferase 3.3 0.005 155 20.56 0.52 S100A8 S100 calcium binding A8 ( A) 3.2 0.004 175 20.11b 0.07b SOD2 SOD2, mitochondrial 3.2 0.001 264 20.61 0.57 ITGB3 Integrin b3 (platelet IIIa, antigen CD61) 3.0 0.006 45 20.74 0.71 NFKBIZ NF of k light polypeptide gene enhancer 3.0 0.004 141 20.67 0.59 in B cells inhibitor, z AKAP12a A kinase (PRKA) anchor protein (gravin) 12 3.0 0.007 57 20.60 0.32b VMP1 Likely ortholog of rat vacuole 1 2.9 0.003 375 20.71 0.59 ADAM9 A disintegrin and metalloproteinase domain 9 (meltrin g) 2.9 0.009 103 20.57 0.41 PTPRC Protein tyrosine phosphatase, receptor type, C 2.9 0.002 56 20.53 0.37b FOSa V-fos FBJ murine osteosarcoma viral oncogene homolog 2.9 0.000 29 20.48 0.49 OSMR Oncostatin M receptor 2.8 0.001 65 20.80 0.73 C9orf71 9 open reading frame 71 2.8 0.005 283 20.41 0.38 EGR1a Early growth response 1 2.8 0.009 184 20.48 0.56 CTSS Cathepsin S 2.7 0.021 190 20.50 0.43 RARRES1 Retinoic acid receptor responder (tazarotene induced) 1 2.7 0.025 76 20.55 0.54 VI2A Ecotropic viral integration site 2A 2.7 0.008 59 20.32b 0.25b Nonredundant and annotated genes are shown. In case of multiple transcripts per gene, the one with highest fold change was selected. Spearman correlations of with eGFR at the time of biopsy and with change in eGFR (DeGFR) at 6 months were considered significant at P,0.05. aPrototypical markers of kidney injury. bP value not significant (P.0.05). were similar to the original set, differing only in longer median kidney function, histology, and IRRAT score expression were time to biopsy (24 versus 16 days), fewer deceased donors confirmed in the validation set biopsies. (30% versus 58%), and fewer grafts with DGF (19% versus The IRRAT score was higher in the original set than in the 31%). Because of these differences, the eGFR was higher in the validation set (2.5 versus 1.6, P=0.02), because the kidneys validation set (31 versus 26 ml/min), reflecting the fact that the from the validation set were less injured (i.e., had better extent of AKI was less than in the original AKI set. In the eGFR and less DGF). Analysis of gene expression revealed validation set, only one graft was lost, due to membranopro- that 19 of the top 30 IRRATs and 212 of the 394 IRRATs were liferative GN, 60 months after the transplant, and one patient significantly increased (FDR=0.05) in the validation set (Sup- died with a functioning graft. plemental Table 1). The correlation of the top 30 IRRATscore in the validation set with the eGFR at biopsy was 20.62 (P,0.001) and with the increase in eGFR by 6 months was 0.45 (P,0.05). The recov- DISCUSSION ery of eGFR was highest in kidneys with high IRRAT scores (Table 7). The acute tubular injury features did not correlate Although all kidney transplantsexperienceAKI, some are more with IRRATscores. Inflammation was again highest in the top injured and require an indication biopsy. When rejection and tertile by the IRRAT score. Thus, the relationships among diseases are excluded, such kidneys constitute a pure AKI

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Table 4. Examples of other IRRATs anticorrelating with eGFR at the time of overlapped the transcripts previously iden- biopsy in AKI biopsies tified in injured mouse kidneys, as well as Correlation many biomarkers previously described in Gene Symbol Gene Title with eGFR body fluids. Thus, the results present a com- TRIM38 Tripartite motif-containing 38 20.82 prehensive picture of molecular events in CLDN1 Claudin 1 20.78 clinically significant human transplant AKI CDC42SE2 CDC42 small effector 2 20.75 that will be useful in supporting diagnostic ICAM1 Intercellular adhesion molecule 1 20.73 applications. IFITM2 IFN induced transmembrane protein 2 (1-8D) 20.73 The elucidation of the transcripts ex- C2CD4A C2 calcium-dependent domain containing 4A 20.73 pressed in human transplant kidneys with 2 KPNA2 Karyopherin a2 (RAG cohort 1, importin a1) 0.72 AKI offers new support to and understand- 2 CASP1 Caspase 1, -related cysteine 0.72 ing of developing biomarkers, refining their peptidase (IL-1, b, convertase) 2 interpretation, and establishing how ex- RASD1 RAS, dexamethasone-induced 1 0.71 fl UGCG UDP-glucose ceramide 20.71 pression of AKI biomarkers in body uids correlates with expression in kidney tissue. TGM2 Transglutaminase 2 (C polypeptide, protein- 20.71 Some biomarkers are more sensitive to glutamine-g-glutamyltransferase) minor injury or more persistent after injury CLIC1 Chloride intracellular channel 1 20.70 resolves, whereas others may be more quan- TFPI Tissue factor pathway inhibitor (lipoprotein- 20.70 titative at distinguishing current functional associated coagulation inhibitor) impairment.22,23 For example, increased CASP8 Caspase 8, apoptosis-related cysteine peptidase 20.69 expression of HAVCR1 is reported in pri- 2 SERPINA1 Serpin peptidase inhibitor, clade A (a-1 0.69 mary diseases24 and in the zero-hour trans- antiproteinase, antitrypsin), member 1 plant biopsies.25 However, in one study, KLF6 Kruppel-like factor 6 20.69 HAVCR1 lacked the ability to distinguish AMACR/// a-methylacyl-CoA racemase///C1q and 20.68 C1QTNF3 TNF-related protein 3 major from minor injury states in kidney 26 TNIK TRAF2 and NCK interacting kinase 20.68 transplants. Moreover, the expression of HN1 Hematologic and neurologic expressed 1 20.67 biomarkers such as HAVCR1 and IL18 in MT1M Metallothionein 1M 20.67 body fluids may also be a sign of systemic PTP4A1 Protein tyrosine phosphatase type IVA, member 1 20.67 trauma.10,27–29 The fact that two AKI, bio- NFKBIZ NF of k light polypeptide gene enhancer 20.67 markers HAVCR1 (KIM1) or IL18 were not in B cells inhibitor, z identified as IRRATs by our initial statistical SEC23IP SEC23 interacting protein 20.67 filtering strategy (fold change and FDR) yet 2 GBP2 Guanylate binding protein 2, IFN-inducible 0.67 were nevertheless associated with AKI phe- 2 MCL1 Myeloid cell leukemia sequence 1 (BCL2-related) 0.66 notype, when re-analyzed, illustrates a ca- MET Met proto-oncogene (hepatocyte growth 20.66 veat about reaching negative conclusions in factor receptor) ABCC4 ATP-binding cassette, subfamily C 20.66 microarray data. Arbitrary selection criteria (CFTR/MRP), member 4 may miss important associations when SFN Stratifin 20.66 many features are highly correlated, and a SOCS3 Suppressor of cytokine signaling 3 20.66 comprehensive examination of the pheno- CPD Carboxypeptidase D 20.66 type associations of all transcripts should be Genes previously identified as top 30 IRRATs by the fold change in AKI biopsies versus pristine PBx included. were excluded because their correlations with eGFR are shown in Table 3. Although IRRAT expression accurately reflected the functional disturbance in cohort with varying degrees of functional impairment. We transplants with AKI, the histologic features believed to reflect compared this cohort to pristine protocol biopsies of similar acute tubular injury did not. Various focal features of acute ages, thus neutralizing the mild injury present even in well tubular necrosis in humans have previously been reported in functioning transplants and identifying the IRRATs as the native kidney AKI,30 the ability of these features to predict signal evoked in human kidneys by recent major parenchymal current GFR impairment or future functional recovery has injury. In the AKI cohort, the IRRATs correlated with a history been poor, indicating that focal abnormalities are not quantita- of DGF and with deceased donors, depression of eGFR at time tively related to overall parenchymal function. The emergence of of biopsy, future eGFR recovery, and interstitial inflammation. the IRRATs as a reliable measurement of AKI in human biopsies Histologic features considered typical of acute tubular injury can be used in place of histopathology, but can also be used to did not correlate with these measurements or with the IRRATs. guide the identification of new histologic features. For exam- An independent validation set confirmed all of the major rela- ple, we recently showed using confocal microscopy that kidney tionships, as well as IRRAT expression. The IRRATs extensively transplants with AKI undetected by conventional features

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Table 5. All 18 identified mouse orthologs of the top 30 The IRRATs show considerable similarity to the transcripts IRRATs show elevated expression in mouse AKI models induced in late kidneys with diseases that predict progression to (isografts and/or ATN) renal failure in such kidneys and that can be used to define a Gene Isografts Isografts ATN IRIT IRIT risk score (e.g., ITGB6, VCAN, NNMT).34 Thus, 11 of the top Symbol and ATN Only Only (I) (L) 30 IRRATs were previously reported by us as frequently used Adamts1 Yes Yes by the molecular risk score classifier for predicting a graft loss Akap12 Yes Yes in late kidney transplant biopsies.34 Although the significance Ctss Yes Yes of this observation is the subject of a separate study, it seems Egr1 Yes likely that the IRRATs are an important signal in a variety of Evi2a Yes renal diseases other than AKI, but their significance will reflect Itgb3 Yes whether that disease is treatable or self-limited or untreatable Fos Yes and relentlessly progressive. In this respect, the close relation- Fcgr4 Yes ship between the top 30 IRRATs and cancer provides insights, Olfm4 Yes Ltf Yes Yes illustrating the ubiquitous nature of the injury-repair/wound- Lcn2 Yes healing response, and recalling the concept of cancers as Nnmt Yes “wounds that do not heal.”35,36 Thus, the IRRATs represent Osmr Yes Yes major general mechanisms such as development37,38 and can- Ptprc Yes cer,37 particularly invasive39 and metastatic cancers,40 as well S100a8 Yes as fibrosis.41,42 Serpina3n Yes Yes Pure AKI in transplants did not lead to late graft failures in Sod2 Yes our analyses, either in the first AKI cohort or the validation set, Vcan Yes a consistent with recent studies indicating that most late kidney Havcr1 Yes Yes transplant failures reflect the onset of diseases such as late Il18a No No ABMR or recurrent disease.16,43 All of the kidneys in the AKI set and the validation set recovered, and the only two kidney aAKI biomarkers that are absent in the human IRRAT set. failures were due to rejection triggered by nonadherence and recurrent GN. Thus, AKI evokes the injury-repair response, like wound healing, and does not lead to progressive deterio- ration in humans. At first glance this may seem to contradict some analyses of DGF in transplant registry data. However, DGF in registry data reflects many different disease states, in- cluding not only AKI but also technical failures and unde- tected ABMR.44,45 Moreover, kidneys with DGF carry the weight of many covariates, such as donor and recipient age, that increase the probability of both DGF and future failure.46 Thus, the impaired long-term survival of kidneys with DGF in registry data are likely not an effect of pure AKI, which is always undesirable but nevertheless inherently reversible. Figure 1. Relationship between IRRAT scores, histopathology, delayed graft function, and eGFR at the time of biopsy for clinical indication in kidneys with AKI. Biopsies (open circles) are ordered CONCISE METHODS by the IRRAT score, and the graph illustrates the relationship of IRRAT scores to the histologic assessment of tubular injury (filled Human Biopsies for Clinical Indications, circles), delayed graft function (*), and eGFR (triangles). Histopathology, and Diagnoses The collection of 234 human kidney biopsies for clinical indications and eight normal nephrectomies was previously described.47 Biopsies have a profound diffuse loss of solute carriers in the brush were obtained under ultrasound guidance by spring-loaded needles border.7 A systematic search for new histologic features that (ASAP Automatic Biopsy; Microvasive, Watertown, MA). This study predict IRRAT expression and GFR is now warranted. More- was approved by the University of Alberta Health Research Ethics over, one histologic feature that correlates with IRRATexpres- Board (Issue #5299), the University of Illinois Chicago Office for sion in kidney transplants with pure AKI, mild interstitial the Protection of Research Subjects (protocol #2006-0544), and the inflammation, should probably be given more weight. Inter- University of Minnesota (protocol HSR#0606 M87646). stitial inflammation is an inherent component of the injury- Biopsies were assessed by a pathologist (M.M.), blinded to the repair response, like the inflammation in a healing wound, and results of molecular studies. The total interstitial inflammation (i total has been described in reperfusion31,32 and DGF.33 score, recorded as a continuous variable48) was scored in both

J Am Soc Nephrol 23: ccc–ccc, 2012 Acute Kidney Injury in Transplants 7 CLINICAL RESEARCH www.jasn.org nonscarred and fibrotic cortical parenchyma. In addition, we assessed to the transplantation process. The mean eGFR in these biopsies was the interstitial fibrosis and tubular atrophy in the parenchyma. We 51.2 compared with a mean of 48.8 in remaining protocol biopsies. previously described the Banff histopathologic diagnoses.47 We thus The histologic features of kidney injury assessed as the percentage analyzed two independent sets of 28 nonrejecting biopsies each with of involved cortical tubules were as follows: loss of brush border, early injury (see Results), 35 biopsies with Banff TCMR, 23 biopsies epithelium-like vacuolization, cytoplasmic lucency, and/or cell ne- with borderline rejection, and 148 remaining biopsies representing crosis. Biopsies with $5% tubules showing such histologic features 46 with ABMR (including 17 C4d-positive and 29 suspicious for were labeled as having acute tubular injury by histology. ABMR, aka C4d-negative ABMR), 10 with mixed (ABMR/TCMR) rejection, 24 with GN, and 68 late nonrejecting biopsies. All diagno- Renal Function ses reflect updated donor-specific antibody status. Renal function was defined by estimated creatinine clearance, using a Protocol biopsies were taken around 6 weeks post-transplant and four-variable Modified Diet in Renal Disease equation at the time of were described previously.14 From those, we selected a subset of biopsies biopsy and 6 months later. We also calculated the change in eGFR at 6 that represented kidneys with a stable future function (at least 2 years of months after the biopsy, and studied the graft losses in the AKI cohort. follow-up), no evidence for AKI and rejection by histology, and no clinical indication for biopsy (clinical or subclinical, before or after RNA Extraction and Microarrays biopsy). Thus, this subset of the pristine protocol biopsies, 11 in total, As previously reported,49 one18-gaugebiopsycorewascollected represents histologically normal kidneys whose only injury pertains for gene expression analysis. RNA extraction, quality control, and HG_U133_Plus_2.0 GeneChip (Affymetrix Santa Clara, CA) processing were described previously.49 Detailed protocols are avail- able in the Affymetrix Technical Manual (www.affymetrix.com).

Microarray Data Analyses Microarray data files from 28 AKI biopsies, 11 protocol biopsies,14 and 8 nephrectomies were preprocessed using robust multichip averaging in Bioconductor and interquartile range filter- ing was used to eliminate probe sets with low variation across the dataset.34 The details of mi- croarray expression data are posted on the Gene Expression Omnibus website (GSE30718). Discovery of transcripts differentially expressed between binary phenotypes (kidneys with AKI and pristine protocol biopsies) of both the test set and the validation set was based on the Welch t test with Benjamini–Hochberg false discovery rate correc- tion(GeneSpring7.3;Agilent,SantaClaraCA).A corrected P value #0.05 was considered significant. Figure 2. Correlation of IRRAT scores (left panel) and AKI scores (right panel) with Summarized expression of transcripts in the eGFR at the time of biopsy (upper panels) or change in eGFR (DeGFR) at 6 months after IRRAT set (set score) was calculated as the biopsy (lower panels) in biopsies for clinical indication with AKI. Values are Spearman geometric mean of the fold change values versus correlation coefficients. AP,0.001; BP.0.05. nephrectomies across all transcripts in a set.

Table 6. Functional annotation of IRRATs in human transplants with AKI by IPA Number of Genes Present Number of Genes Present Annotation Tool Function or Pathway in All 394 IRAATs in the Top 30 IRRATs (% of All IRRATs) (% of Top 30 IRRATs) IPA diseases and disorders Cancer (n=6087) 215 (55) 25 (83) IPA molecular and cellular function Cellular movement (n=2755) 131 (33) 19 (63) IPA development and function Tissue development (n=959) 100 (25) 11 (36) IPA canonical pathway Acute phase response signaling (n=172) 19 (5) 4 (13) All IRRATs $1.5-fold in AKI versus protocol biopsies (394, nonredundant) and 30 top IRRATs were used for annotation. Enrichment in Ingenuity Pathway Analysis (IPA) terms was tested by the Fisher’s exact test. All enrichments were highly significant, P,0.0001, and remained significant after the multiple test correction (P values ranged from 0.023 to ,0.0001).

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Statistical Analyses Statistical significance of differences between the groups of biopsies was tested by Kruskal– Wallis test with Dunn’s correction for multiple comparisons. The Spearman two-tailed correla- tion was used to analyze relationship between features and gene set scores. The chi-squared test was used for contingency tables. P values ,0.05 were considered significant. AUC analy- sis tested the ability of transcripts to predict AKI.

ACKNOWLEDGMENTS

The authors thank Anna Hutton and Vido Ramassar for providing technical support and Zija Jacaj for collecting the clinical data. The authors also thank Dr. Bruce Kaplan and Dr. Arthur Matas for biopsy material. The Alberta Transplant Applied Genomic Centre has been supported by Genome Canada, University of Alberta, University of Alberta Hospital Foundation, Alberta Advanced Edu- Figure 3. Prediction of AKI by individual transcripts. (A) Comparison of AKI biopsies to cation and Technology, Roche Molecular Sys- pristine protocol biopsies. All transcripts that went into the analysis are shown. (B) Only tems, Hoffmann-La Roche Canada Ltd, Alberta the top 30 IRRATs, HAVCR1, and IL18 are shown. (C) Comparison of AKI biopsies to Ministry of Advanced Education and Technol- pristine protocol biopsies. Only transcripts significant at the false discovery rate of 0.05 ogy, Roche Organ Transplant Research Foun- are shown. (D) Vertical line indicates the uncorrected P value of 0.05. The top 30 IR- RATs are represented by yellow dots, and HAVCR1 and IL18 by pink dots. dation (grants to B.S. and M.M.), Kidney Foundation of Canada, Stromedix Inc, and Astellas Canada. P.F.H. also holds a Canada Re- Table 7. Relationship of the IRRAT score to the histology search Chair in Transplant Immunology and the Muttart Chair in and function of kidneys with AKI in the validation set Clinical Immunology. Bottom Middle Top Tertile Tertile (n=9) Tertile (n=9) (n=10) eGFR at biopsya 39.8b 31.3 23.2 DISCLOSURES c c eGFR at 6 mo 47.2 50.5 47.7 P.F.H. holds shares in Transcriptome Sciences Inc, a company with an b DeGFR by 6 mo 7.4 20.1 24.9 interest in molecular diagnostics. The other authors have no competing Features of ATI (yes/no) 4/5 7/2 7/3 financial interests. i-total (%) 6.3b 2.6d 18.8 Delayed/immediate 0/9 2/5 3/6 e graft function REFERENCES Deceased/living donor 2/7 3/5 3/7 Kruskal–Wallis test with Dunn’s correction for multiple comparisons. The top 1. Venkatachalam MA, Griffin KA, Lan RP, Geng H, Saikumar P, Bidani AK: tertile was compared with the remaining tertiles. Graft function and the donor type are represented by one biopsy per patient. Acute kidney injury: A springboard for progression in chronic kidney – aeGFR and DeGFR are in milliliters per minute per 1.73 m2. disease. Am J Physiol Renal Physiol 298: F1078 F1094, 2010 bP,0.01. 2. Sharfuddin AA, Molitoris BA: Pathophysiology of ischemic acute kidney cIndicates significant difference between eGFR at biopsy and 6-month eGFR, injury. Nat Rev Nephrol 7: 189–200, 2011 by the paired t test. 3. Devarajan P: Update on mechanisms of ischemic acute kidney injury. d P,0.001. J Am Soc Nephrol 17: 1503–1520, 2006 e Missing data for two grafts. 4. Siew ED, Ware LB, Ikizler TA: Biological markers of acute kidney injury. J Am Soc Nephrol 22: 810–820, 2011 Functional annotations of transcript sets were done using the 5. Famulski KS, Broderick G, Einecke G, Hay K, Cruz J, Sis B, Mengel M, Halloran PF: Transcriptome analysis reveals heterogeneity in the injury Ingenuity Pathway Analysis Global Molecular Network. Enrichment response of kidney transplants. Am J Transplant 7: 2483–2495, 2007 ’ in functional categories was tested by Fisher s exact test. A P value 6. Edemir B, Kurian SM, Eisenacher M, Lang D, Müller-Tidow C, ,0.05 was considered significant. Gabriëls G, Salomon DR, Schlatter E: Activation of counter-

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J Am Soc Nephrol 23: ccc–ccc, 2012 Acute Kidney Injury in Transplants 11 ID AKI vs control P value Symbol Gene Name *In case of multiple probesets per gene, one with the highest fold change was selected. Fold Change 208083_s_at 7.88 0.000932 ITGB6 integrin, beta 6 202376_at 6.12 0.000518 SERPINA3 serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 3 1553575_at 5.62 0.0033 MT-ND6 NADH dehydrogenase, subunit 6 (complex I) 212768_s_at 5.50 0.000896 OLFM4 olfactomedin 4 206157_at 5.26 0.00177 PTX3 pentraxin 3, long 212531_at 4.26 0.00405 LCN2 lipocalin 2 215646_s_at 4.13 0.00408 VCAN versican 202018_s_at 4.12 0.0318 LTF lactotransferrin 203021_at 4.05 0.0129 SLPI secretory leukocyte peptidase inhibitor 222486_s_at 4.03 0.000329 ADAMTS1 ADAM metallopeptidase with thrombospondin type 1 motif, 1 1552439_s_at 3.82 0.000714 MEGF11 multiple EGF-like-domains 11 210602_s_at 3.74 0.000408 CDH6 cadherin 6, type 2, K-cadherin (fetal kidney) 229947_at 3.62 0.00843 PI15 peptidase inhibitor 15 204006_s_at 3.39 0.00241 FCGR3A Fc fragment of IgG, low affinity IIIa, receptor (CD16a) 202238_s_at 3.29 0.00492 NNMT nicotinamide N-methyltransferase 202917_s_at 3.20 0.00369 S100A8 S100 calcium binding protein A8 215223_s_at 3.17 0.000516 SOD2 superoxide dismutase 2, mitochondrial 204627_s_at 3.04 0.00619 ITGB3 integrin, beta 3 (platelet glycoprotein IIIa, antigen CD61) 223217_s_at 2.99 0.00397 NFKBIZ nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, zeta 231067_s_at 2.97 0.00681 AKAP12 A kinase (PRKA) anchor protein 12 224917_at 2.94 0.00256 VMP1/ mir-21likely ortholog of rat vacuole membrane protein 1/ microRNA 21 1555326_a_at 2.93 0.00891 ADAM9 ADAM metallopeptidase domain 9 207238_s_at 2.90 0.0021 PTPRC protein tyrosine phosphatase, receptor type, C 209189_at 2.89 8.30E-05 FOS FBJ murine osteosarcoma viral oncogene homolog 205729_at 2.82 0.000797 OSMR oncostatin M receptor 1553809_a_at 2.82 0.00485 C9orf71 chromosome 9 open reading frame 71 227404_s_at 2.79 0.00858 EGR1 early growth response 1 202902_s_at 2.70 0.0206 CTSS cathepsin S 206392_s_at 2.68 0.0251 RARRES1 retinoic acid receptor responder (tazarotene induced) 1 204774_at 2.66 0.00821 EVI2A ecotropic viral integration site 2A 232654_s_at 2.63 0.00112 UGT1A6 UDP 1 family, polypeptide A6 216598_s_at 2.61 0.0254 CCL2 chemokine (C-C motif) ligand 2 221577_x_at 2.58 0.000964 GDF15 growth differentiation factor 15 221765_at 2.57 0.00485 UGCG UDP-glucose ceramide glucosyltransferase 226142_at 2.57 0.0105 GLIPR1 GLI pathogenesis-related 1 201720_s_at 2.56 0.0101 LAPTM5 lysosomal protein transmembrane 5 1555728_a_at 2.53 0.00772 MS4A4A membrane-spanning 4-domains, subfamily A, member 4 209774_x_at 2.53 0.0181 CXCL2 chemokine (C-X-C motif) ligand 2 1569003_at 2.52 0.00205 TMEM49 transmembrane protein 49 1555229_a_at 2.51 0.00681 C1S complement component 1, s subcomponent 216450_x_at 2.45 0.00172 HSP90B1 heat shock protein 90kDa beta (Grp94), member 1 1553530_a_at 2.45 0.0129 ITGB1 integrin, beta 1 (fibronectin receptor, beta polypeptide, antigen CD29 includes MDF2, MSK12) 241031_at 2.44 0.00551 C2CD4A C2 calcium-dependent domain containing 4A 201942_s_at 2.39 0.000792 CPD carboxypeptidase D 217546_at 2.38 0.0283 MT1M metallothionein 1M 211742_s_at 2.38 0.0167 EVI2B ecotropic viral integration site 2B 219558_at 2.37 0.000704 ATP13A3 ATPase type 13A3 235281_x_at 2.36 0.00403 AHNAK AHNAK nucleoprotein 204864_s_at 2.36 0.00112 IL6ST interleukin 6 signal transducer (gp130, oncostatin M receptor) 1555564_a_at 2.35 0.00202 CFI complement factor I 225987_at 2.35 0.0225 STEAP4 STEAP family member 4 1558015_s_at 2.35 0.00376 ACTR2 ARP2 actin-related protein 2 homolog (yeast) 1563874_at 2.34 0.00779 WDR72 WD repeat domain 72 204959_at 2.32 0.0025 MNDA myeloid cell nuclear differentiation antigen 1555349_a_at 2.31 0.0325 ITGB2 integrin, beta 2 (complement component 3 receptor 3 and 4 subunit) 228222_at 2.29 0.00407 PPP1CB 1, catalytic subunit, beta isozyme 201666_at 2.29 0.0024 TIMP1 TIMP metallopeptidase inhibitor 1 214336_s_at 2.28 0.0011 COPA coatomer protein complex, subunit alpha 1558214_s_at 2.28 0.000377 CTNNA1 catenin (cadherin-associated protein), alpha 1, 102kDa 223467_at 2.27 0.0066 RASD1 RAS, dexamethasone-induced 1 215049_x_at 2.26 0.0481 CD163 CD163 molecule 206584_at 2.25 0.0101 LY96 lymphocyte antigen 96 200671_s_at 2.23 0.00237 SPTBN1 spectrin, beta, non-erythrocytic 1 215719_x_at 2.23 0.0415 FAS Fas (TNF receptor superfamily, member 6) 208621_s_at 2.23 0.000167 EZR ezrin 206211_at 2.23 0.0155 SELE E 1555996_s_at 2.22 0.0135 EIF4A2 eukaryotic translation initiation factor 4A2 209676_at 2.20 0.0246 TFPI tissue factor pathway inhibitor (lipoprotein-associated coagulation inhibitor) 210432_s_at 2.18 0.0213 SCN3A sodium channel, voltage-gated, type III, alpha subunit 229622_at 2.18 0.0379 FAM132B family with sequence similarity 132, member B 201858_s_at 2.16 0.00204 SRGN serglycin 201211_s_at 2.16 0.00509 DDX3X DEAD (Asp-Glu-Ala-Asp) box polypeptide 3, X-linked 201890_at 2.16 0.00875 RRM2 ribonucleotide reductase M2 201101_s_at 2.16 0.00297 BCLAF1 BCL2-associated transcription factor 1 1555167_s_at 2.15 0.00522 NAMPT nicotinamide phosphoribosyltransferase 200796_s_at 2.15 0.000857 MCL1 myeloid cell leukemia sequence 1 (BCL2-related) 212671_s_at 2.14 0.0275 HLA-DQA1 major histocompatibility complex, class II, DQ alpha 1 210148_at 2.14 0.0475 HIPK3 homeodomain interacting 3 227697_at 2.14 0.0479 SOCS3 suppressor of cytokine signaling 3 217465_at 2.13 0.000797 NCKAP1 NCK-associated protein 1 212105_s_at 2.13 0.000129 DHX9 DEAH (Asp-Glu-Ala-His) box polypeptide 9 202357_s_at 2.13 0.0049 CFB complement factor B 233827_s_at 2.12 0.00132 SUPT16H suppressor of Ty 16 homolog (S. cerevisiae) 222387_s_at 2.12 0.00249 VPS35 vacuolar protein sorting 35 homolog (S. cerevisiae) 1554878_a_at 2.11 0.035 ABCD3 ATP-binding cassette, sub-family D (ALD), member 3 211981_at 2.10 0.0022 COL4A1 collagen, type IV, alpha 1 234989_at 2.10 0.0162 NEAT1 nuclear assembly transcript 1 (non-protein coding) 230318_at 2.10 0.0011 SERPINA1 serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 1 203535_at 2.09 0.000574 S100A9 S100 calcium binding protein A9 201399_s_at 2.09 0.0126 TRAM1 translocation associated membrane protein 1 206552_s_at 2.08 0.0181 TAC1 tachykinin, precursor 1 211599_x_at 2.07 0.000277 MET met proto-oncogene (hepatocyte growth factor receptor) 222549_at 2.06 0.0209 CLDN1 claudin 1 204971_at 2.06 0.0209 CSTA cystatin A (stefin A) 202206_at 2.04 0.0155 ARL4C ADP-ribosylation factor-like 4C 202604_x_at 2.03 0.011 ADAM10 ADAM metallopeptidase domain 10 222608_s_at 2.03 0.00444 ANLN anillin, actin binding protein 200641_s_at 2.03 0.0131 YWHAZ tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta polypeptide 205174_s_at 2.02 0.0234 QPCT glutaminyl-peptide cyclotransferase 210895_s_at 2.02 0.0026 CD86 CD86 molecule 208727_s_at 2.01 0.0405 CDC42 cell division cycle 42 (GTP binding protein, 25kDa) 211395_x_at 2.01 0.000356 FCGR2C Fc fragment of IgG, low affinity IIc, receptor for (CD32) (gene/) 205476_at 2.00 0.0442 CCL20 chemokine (C-C motif) ligand 20 33323_r_at 2.00 0.00631 SFN stratifin 204983_s_at 2.00 0.00669 GPC4 4 212298_at 2.00 0.0284 NRP1 neuropilin 1 1555434_a_at 1.99 0.000126 SLC39A14 solute carrier family 39 (zinc transporter), member 14 224356_x_at 1.98 0.0108 MS4A6A membrane-spanning 4-domains, subfamily A, member 6A 220585_at 1.98 0.000493 HKDC1 hexokinase domain containing 1 232591_s_at 1.98 0.0181 TMEM30A transmembrane protein 30A 206207_at 1.97 0.000516 CLC Charcot-Leyden crystal protein 219508_at 1.97 0.00477 GCNT3 glucosaminyl (N-acetyl) 3, type 202075_s_at 1.97 0.0203 PLTP phospholipid transfer protein 1552613_s_at 1.96 0.000968 CDC42SE2 CDC42 small effector 2 213446_s_at 1.96 3.97E-05 IQGAP1 IQ motif containing GTPase activating protein 1 1567628_at 1.96 0.0121 CD74 CD74 molecule, major histocompatibility complex, class II invariant chain 204787_at 1.95 0.0348 VSIG4 V-set and immunoglobulin domain containing 4 214467_at 1.95 0.02 GPR65 -coupled receptor 65 200841_s_at 1.94 0.000239 EPRS glutamyl-prolyl-tRNA synthetase 1554029_a_at 1.94 0.00365 TTC37 tetratricopeptide repeat domain 37 209955_s_at 1.94 0.0482 FAP fibroblast activation protein, alpha 1552611_a_at 1.94 1.31E-05 JAK1 Janus kinase 1 210559_s_at 1.93 0.00176 CDK1 -dependent kinase 1 1555203_s_at 1.93 0.00484 SLC44A4 solute carrier family 44, member 4 203305_at 1.92 0.0308 F13A1 coagulation factor XIII, A1 polypeptide 213548_s_at 1.92 0.00089 CDV3 CDV3 homolog (mouse) 212806_at 1.92 0.0209 PRUNE2 prune homolog 2 (Drosophila) 215891_s_at 1.92 0.0248 GM2A GM2 ganglioside activator 212797_at 1.91 0.00525 SORT1 sortilin 1 1568574_x_at 1.91 0.0273 SPP1 secreted phosphoprotein 1 204213_at 1.90 0.0078 PIGR polymeric immunoglobulin receptor 213629_x_at 1.90 0.0135 MT1F metallothionein 1F 217523_at 1.89 0.00464 CD44 CD44 molecule (Indian blood group) 214845_s_at 1.89 0.00107 CALU calumenin 202503_s_at 1.89 0.0373 KIAA0101 KIAA0101 227260_at 1.88 0.0485 ANKRD10 ankyrin repeat domain 10 211966_at 1.88 0.00914 COL4A2 collagen, type IV, alpha 2 205650_s_at 1.88 0.00155 FGA fibrinogen alpha chain 210904_s_at 1.88 0.000263 IL13RA1 interleukin 13 receptor, alpha 1 219148_at 1.87 0.0327 PBK PDZ binding kinase 205082_s_at 1.87 0.0339 AOX1 aldehyde oxidase 1 206396_at 1.87 0.00217 SLC1A1 solute carrier family 1 (neuronal/epithelial high affinity glutamate transporter, system Xag), member 1 211681_s_at 1.87 0.00129 PDLIM5 PDZ and LIM domain 5 1553171_x_at 1.86 0.0322 LRRN4 leucine rich repeat neuronal 4 201107_s_at 1.86 1.63E-05 THBS1 thrombospondin 1 202859_x_at 1.86 0.0193 IL8 interleukin 8 210830_s_at 1.86 0.0263 PON2 paraoxonase 2 202628_s_at 1.86 0.0387 SERPINE1 serpin peptidase inhibitor, clade E (nexin, plasminogen activator inhibitor type 1), member 1 208853_s_at 1.85 0.011 CANX 206632_s_at 1.85 0.0121 APOBEC3B apolipoprotein B mRNA editing , catalytic polypeptide-like 3B 210971_s_at 1.85 0.00731 ARNTL aryl hydrocarbon receptor nuclear translocator-like 1553962_s_at 1.85 0.043 RHOB ras homolog gene family, member B 209641_s_at 1.84 0.0157 ABCC3 ATP-binding cassette, sub-family C (CFTR/MRP), member 3 209424_s_at 1.84 0.0027 AMACR alpha-methylacyl-CoA racemase 223344_s_at 1.84 0.0109 MS4A7 membrane-spanning 4-domains, subfamily A, member 7 210935_s_at 1.84 0.000327 WDR1 WD repeat domain 1 228188_at 1.84 0.0292 FOSL2 FOS-like antigen 2 208965_s_at 1.84 0.0437 IFI16 interferon, gamma-inducible protein 16 214701_s_at 1.84 0.0026 FN1 fibronectin 1 206116_s_at 1.84 0.00986 TPM1 tropomyosin 1 (alpha) 206917_at 1.84 0.00475 GNA13 guanine nucleotide binding protein (G protein), alpha 13 210337_s_at 1.84 0.00056 ACLY ATP citrate lyase 203736_s_at 1.82 0.041 PPFIBP1 PTPRF interacting protein, binding protein 1 (liprin beta 1) 222712_s_at 1.82 0.0281 MUC13 mucin 13, cell surface associated 226789_at 1.81 0.0137 EMB embigin 203828_s_at 1.81 0.0258 IL32 interleukin 32 210892_s_at 1.81 0.00132 GTF2I general transcription factor IIi 229296_at 1.81 0.0197 LOC100128501 hypothetical protein LOC100128501 221960_s_at 1.81 0.00111 RAB2A RAB2A, member RAS oncogene family 209257_s_at 1.81 0.00742 SMC3 structural maintenance of 3 202431_s_at 1.80 0.0371 MYC v-myc myelocytomatosis viral oncogene homolog (avian) 1553749_at 1.80 0.00785 FAM76B family with sequence similarity 76, member B 213160_at 1.80 0.0235 DOCK2 dedicator of cytokinesis 2 208885_at 1.79 0.0256 LCP1 lymphocyte cytosolic protein 1 (L-plastin) 1554241_at 1.79 0.00883 COCH coagulation factor C homolog, cochlin (Limulus polyphemus) 208581_x_at 1.79 0.0274 MT1X metallothionein 1X 208309_s_at 1.79 0.0176 MALT1 mucosa associated lymphoid tissue lymphoma translocation gene 1 221220_s_at 1.79 0.0336 SCYL2 SCY1-like 2 (S. cerevisiae) 238419_at 1.77 0.0011 PHLDB2 pleckstrin homology-like domain, family B, member 2 202236_s_at 1.77 0.0286 SLC16A1 solute carrier family 16, member 1 (monocarboxylic acid transporter 1) 209114_at 1.77 0.000859 TSPAN1 tetraspanin 1 214560_at 1.76 0.0137 FPR3 formyl peptide receptor 3 211354_s_at 1.76 0.00376 LEPR leptin receptor 218755_at 1.76 0.00684 KIF20A family member 20A 225647_s_at 1.76 0.00127 CTSC cathepsin C 216915_s_at 1.76 0.0197 PTPN12 protein tyrosine phosphatase, non-receptor type 12 1567213_at 1.75 0.0225 PNN pinin, associated protein 201559_s_at 1.75 0.0234 CLIC4 chloride intracellular channel 4 214040_s_at 1.75 0.00295 GSN 210982_s_at 1.75 0.00945 HLA-DRA major histocompatibility complex, class II, DR alpha 1554508_at 1.75 0.000466 PIK3AP1 phosphoinositide-3-kinase adaptor protein 1 216392_s_at 1.75 0.0106 SEC23IP SEC23 interacting protein 202638_s_at 1.75 0.0235 ICAM1 intercellular adhesion molecule 1 204426_at 1.75 0.0373 TMED2 transmembrane emp24 domain trafficking protein 2 210764_s_at 1.74 0.0454 CYR61 cysteine-rich, angiogenic inducer, 61 220483_s_at 1.74 0.000632 RNF19A ring finger protein 19A 210732_s_at 1.74 0.000481 LGALS8 lectin, galactoside-binding, soluble, 8 215220_s_at 1.74 0.0018 TPR translocated promoter region (to activated MET oncogene) 207029_at 1.74 0.00365 KITLG KIT ligand 238469_at 1.73 0.00855 OGFRL1 opioid growth factor receptor-like 1 209024_s_at 1.73 0.00984 SYNCRIP binding, cytoplasmic RNA interacting protein 1555039_a_at 1.73 0.00217 ABCC4 ATP-binding cassette, sub-family C (CFTR/MRP), member 4 211016_x_at 1.73 0.00489 HSPA4 heat shock 70kDa protein 4 228648_at 1.73 3.97E-05 LRG1 leucine-rich alpha-2-glycoprotein 1 210087_s_at 1.73 0.000558 MPZL1 -like 1 203294_s_at 1.73 0.0228 LMAN1 lectin, mannose-binding, 1 212185_x_at 1.73 0.00954 MT2A metallothionein 2A 208664_s_at 1.73 0.0437 TTC3 tetratricopeptide repeat domain 3 206420_at 1.73 0.00659 IGSF6 immunoglobulin superfamily, member 6 1557910_at 1.73 0.005 HSP90AB1 heat shock protein 90kDa alpha (cytosolic), class B member 1 216336_x_at 1.73 0.0172 MT1E metallothionein 1E 200607_s_at 1.72 0.0131 RAD21 RAD21 homolog (S. pombe) 212257_s_at 1.72 0.00382 SMARCA2 SWI/SNF related, matrix associated, actin dependent regulator of , subfamily a, member 2 206582_s_at 1.72 0.016 GPR56 G protein-coupled receptor 56 201647_s_at 1.72 0.0403 SCARB2 scavenger receptor class B, member 2 209131_s_at 1.72 0.0305 SNAP23 synaptosomal-associated protein, 23kDa 202748_at 1.71 0.00498 GBP2 guanylate binding protein 2, interferon-inducible 1557418_at 1.71 0.00666 ACSL4 acyl-CoA synthetase long-chain family member 4 207549_x_at 1.71 0.0202 CD46 CD46 molecule, complement regulatory protein 212909_at 1.71 0.0202 LYPD1 LY6/PLAUR domain containing 1 204344_s_at 1.71 0.0018 SEC23A Sec23 homolog A (S. cerevisiae) 204192_at 1.70 0.0417 CD37 CD37 molecule 1554747_a_at 1.70 0.00619 SEPT2 septin 2 1552703_s_at 1.70 0.00811 CASP1 caspase 1, apoptosis-related cysteine peptidase (interleukin 1, beta, convertase) 1553565_s_at 1.70 0.00349 DDAH1 dimethylarginine dimethylaminohydrolase 1 1564494_s_at 1.70 0.000695 P4HB prolyl 4-hydroxylase, beta polypeptide 210052_s_at 1.70 0.000224 TPX2 TPX2, microtubule-associated, homolog (Xenopus laevis) 203158_s_at 1.70 0.03 GLS glutaminase 221667_s_at 1.70 0.00801 HSPB8 heat shock 22kDa protein 8 1554375_a_at 1.69 0.0159 NR1H4 nuclear receptor subfamily 1, group H, member 4 201596_x_at 1.69 0.00445 KRT18 keratin 18 1552261_at 1.68 0.000516 WFDC2 WAP four-disulfide core domain 2 203676_at 1.68 0.00383 GNS glucosamine (N-acetyl)-6-sulfatase 206461_x_at 1.68 0.0344 MT1H metallothionein 1H 214456_x_at 1.68 0.0131 SAA1 serum amyloid A1 213418_at 1.68 0.0399 HSPA6 heat shock 70kDa protein 6 (HSP70B') 218368_s_at 1.68 0.00411 TNFRSF12A receptor superfamily, member 12A 235549_at 1.68 0.0223 RNF144B ring finger protein 144B 1555756_a_at 1.67 0.0186 CLEC7A C-type lectin domain family 7, member A 215910_s_at 1.67 0.00804 FNDC3A fibronectin type III domain containing 3A 1556551_s_at 1.67 0.0203 SLC39A6 solute carrier family 39 (zinc transporter), member 6 214511_x_at 1.67 0.0422 FCGR1B Fc fragment of IgG, high affinity Ib, receptor (CD64) 1558733_at 1.67 0.00132 ZBTB38 zinc finger and BTB domain containing 38 204136_at 1.67 0.00623 COL7A1 collagen, type VII, alpha 1 201292_at 1.67 0.0323 TOP2A topoisomerase (DNA) II alpha 170kDa 204007_at 1.67 0.0274 FCGR3B Fc fragment of IgG, low affinity IIIb, receptor (CD16b) 206034_at 1.67 1.42E-05 SERPINB8 serpin peptidase inhibitor, clade B (ovalbumin), member 8 224149_x_at 1.67 0.0202 SLMAP sarcolemma associated protein 219161_s_at 1.67 0.0171 CKLF chemokine-like factor 201008_s_at 1.66 0.0349 TXNIP thioredoxin interacting protein 215936_s_at 1.66 0.0473 KIAA1033 KIAA1033 205237_at 1.66 0.018 FCN1 ficolin (collagen/fibrinogen domain containing) 1 225412_at 1.66 0.0384 TMEM87B transmembrane protein 87B 217755_at 1.66 0.00098 HN1 hematological and neurological expressed 1 222385_x_at 1.66 0.0023 SEC61A1 Sec61 alpha 1 subunit (S. cerevisiae) 205334_at 1.66 0.0227 S100A1 S100 calcium binding protein A1 205017_s_at 1.65 0.00129 MBNL2 muscleblind-like 2 (Drosophila) 226825_s_at 1.65 0.000154 TMEM165 transmembrane protein 165 219918_s_at 1.65 0.00361 ASPM asp (abnormal spindle) homolog, microcephaly associated (Drosophila) 206924_at 1.65 0.0086 IL11 interleukin 11 201946_s_at 1.65 0.00134 CCT2 chaperonin containing TCP1, subunit 2 (beta) 205266_at 1.64 0.00766 LIF leukemia inhibitory factor (cholinergic differentiation factor) 1552978_a_at 1.64 0.0478 SCAMP1 secretory carrier membrane protein 1 216942_s_at 1.64 0.0205 CD58 CD58 molecule 211456_x_at 1.64 0.0165 MT1P2 metallothionein 1 pseudogene 2 217208_s_at 1.64 0.000696 DLG1 discs, large homolog 1 (Drosophila) 221078_s_at 1.64 0.000428 CCDC88A coiled-coil domain containing 88A 200776_s_at 1.64 0.0212 BZW1 basic leucine zipper and W2 domains 1 201538_s_at 1.64 0.00379 DUSP3 dual specificity phosphatase 3 223809_at 1.64 0.000672 RGS18 regulator of G-protein signaling 18 204745_x_at 1.64 0.0332 MT1G metallothionein 1G 214786_at 1.63 0.0414 MAP3K1 mitogen-activated protein kinase kinase kinase 1 203610_s_at 1.63 0.00266 TRIM38 tripartite motif-containing 38 205446_s_at 1.63 0.00989 ATF2 activating transcription factor 2 200604_s_at 1.63 0.00384 PRKAR1A protein kinase, cAMP-dependent, regulatory, type I, alpha (tissue specific extinguisher 1) 208629_s_at 1.63 0.000235 HADHA hydroxyacyl-CoA dehydrogenase/3-ketoacyl-CoA thiolase/enoyl-CoA hydratase (trifunctional protein), alpha subunit 1565483_at 1.62 0.002 EGFR epidermal growth factor receptor 202884_s_at 1.62 0.004 PPP2R1B , regulatory subunit A, beta 217294_s_at 1.62 0.00431 ENO1 enolase 1, (alpha) 1553944_at 1.62 0.0181 MIA2 melanoma inhibitory activity 2 208699_x_at 1.62 0.000493 TKT transketolase 216950_s_at 1.61 0.00675 FCGR1A Fc fragment of IgG, high affinity Ia, receptor (CD64) 208633_s_at 1.61 0.00743 MACF1 microtubule-actin crosslinking factor 1 213674_x_at 1.61 0.0223 IGHD immunoglobulin heavy constant delta 233573_s_at 1.61 0.00373 WDR6 WD repeat domain 6 202444_s_at 1.61 0.00385 ERLIN1 ER lipid raft associated 1 211652_s_at 1.61 0.00374 LBP lipopolysaccharide binding protein 210827_s_at 1.60 0.0135 ELF3 E74-like factor 3 (ets domain transcription factor, epithelial-specific ) 214168_s_at 1.60 0.0428 TJP1 tight junction protein 1 (zona occludens 1) 206111_at 1.60 4.57E-05 RNASE2 ribonuclease, RNase A family, 2 (liver, eosinophil-derived neurotoxin) 201070_x_at 1.60 0.00766 SF3B1 splicing factor 3b, subunit 1, 155kDa 201971_s_at 1.60 0.0408 ATP6V1A ATPase, H+ transporting, lysosomal 70kDa, V1 subunit A 1555419_a_at 1.60 0.0385 ASAH1 N-acylsphingosine amidohydrolase (acid ceramidase) 1 218729_at 1.60 0.04 LXN latexin 1557100_s_at 1.59 0.00801 HECTD1 HECT domain containing 1 219978_s_at 1.59 4.56E-05 NUSAP1 nucleolar and spindle associated protein 1 227214_at 1.59 0.00497 GOPC golgi-associated PDZ and coiled-coil motif containing 221638_s_at 1.59 0.0105 STX16 syntaxin 16 204846_at 1.59 0.00819 CP ceruloplasmin (ferroxidase) 233924_s_at 1.59 0.00295 EXOC6 exocyst complex component 6 204502_at 1.59 0.00288 SAMHD1 SAM domain and HD domain 1 208791_at 1.59 0.0173 CLU 206113_s_at 1.58 0.0346 RAB5A RAB5A, member RAS oncogene family 204470_at 1.58 0.0261 CXCL1 chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha) 203015_s_at 1.58 0.016 SSX2IP synovial sarcoma, X breakpoint 2 interacting protein 222646_s_at 1.58 0.0186 ERO1L ERO1-like (S. cerevisiae) 214875_x_at 1.58 6.49E-05 APLP2 amyloid beta (A4) precursor-like protein 2 202267_at 1.58 0.0326 LAMC2 laminin, gamma 2 1552264_a_at 1.58 0.001 MAPK1 mitogen-activated protein kinase 1 200769_s_at 1.57 0.043 MAT2A methionine adenosyltransferase II, alpha 211762_s_at 1.57 1.63E-05 KPNA2 karyopherin alpha 2 (RAG cohort 1, importin alpha 1) 208983_s_at 1.57 0.00669 PECAM1 platelet/endothelial molecule 1554614_a_at 1.57 0.025 PTBP2 polypyrimidine tract binding protein 2 1555830_s_at 1.57 0.00508 ESYT2 extended synaptotagmin-like protein 2 1555758_a_at 1.57 0.0209 CDKN3 cyclin-dependent kinase inhibitor 3 206788_s_at 1.57 0.00105 CBFB core-binding factor, beta subunit 215236_s_at 1.57 0.0477 PICALM phosphatidylinositol binding clathrin assembly protein 238421_at 1.57 0.0299 RC3H2 ring finger and CCCH-type domains 2 201954_at 1.57 0.0268 ARPC1B actin related protein 2/3 complex, subunit 1B, 41kDa 217881_s_at 1.57 0.0274 CDC27 cell division cycle 27 homolog (S. cerevisiae) 215633_x_at 1.57 0.00143 LST1 leukocyte specific transcript 1 206372_at 1.57 0.0344 MYF6 myogenic factor 6 (herculin) 213906_at 1.56 0.036 MYBL1 v-myb myeloblastosis viral oncogene homolog (avian)-like 1 213226_at 1.56 0.0374 CCNA2 cyclin A2 205193_at 1.56 0.00172 MAFF v-maf musculoaponeurotic fibrosarcoma oncogene homolog F (avian) 1554600_s_at 1.56 0.000854 LMNA lamin A/C 206383_s_at 1.56 0.00756 G3BP2 GTPase activating protein (SH3 domain) binding protein 2 208879_x_at 1.56 0.00801 PRPF6 PRP6 pre-mRNA processing factor 6 homolog (S. cerevisiae) 211656_x_at 1.55 0.0387 HLA-DQB1 major histocompatibility complex, class II, DQ beta 1 1555594_a_at 1.55 0.00231 MBNL1 muscleblind-like (Drosophila) 202241_at 1.55 0.0317 TRIB1 tribbles homolog 1 (Drosophila) 214196_s_at 1.55 0.0352 TPP1 tripeptidyl peptidase I 201044_x_at 1.55 0.000157 DUSP1 dual specificity phosphatase 1 208744_x_at 1.55 0.00619 HSPH1 heat shock 105kDa/110kDa protein 1 200889_s_at 1.55 0.00388 SSR1 signal sequence receptor, alpha 223333_s_at 1.55 0.00317 ANGPTL4 angiopoietin-like 4 1555812_a_at 1.55 0.0101 ARHGDIB Rho GDP dissociation inhibitor (GDI) beta 210815_s_at 1.55 0.0243 CALCRL calcitonin receptor-like 1552660_a_at 1.55 0.0373 C5orf22 open reading frame 22 209653_at 1.55 0.0477 KPNA4 karyopherin alpha 4 (importin alpha 3) 1559954_s_at 1.54 0.0155 DDX42 DEAD (Asp-Glu-Ala-Asp) box polypeptide 42 211488_s_at 1.54 0.00437 ITGB8 integrin, beta 8 236783_at 1.54 0.015 KCNIP4 Kv channel interacting protein 4 211450_s_at 1.54 0.0463 MSH6 mutS homolog 6 (E. coli) 213461_at 1.54 0.0399 NUDT21 nudix (nucleoside diphosphate linked moiety X)-type motif 21 208721_s_at 1.54 0.00147 ANAPC5 anaphase promoting complex subunit 5 1569616_at 1.54 0.000718 GEMC1 geminin coiled-coil domain-containing protein 1 202870_s_at 1.54 0.00772 CDC20 cell division cycle 20 homolog (S. cerevisiae) 1555240_s_at 1.54 0.00112 GNG12 guanine nucleotide binding protein (G protein), gamma 12 212022_s_at 1.54 0.000213 MKI67 antigen identified by monoclonal antibody Ki-67 243358_at 1.53 0.0132 IGF1R insulin-like growth factor 1 receptor 211661_x_at 1.53 0.000671 PTAFR platelet-activating factor receptor 202873_at 1.53 0.0112 ATP6V1C1 ATPase, H+ transporting, lysosomal 42kDa, V1 subunit C1 222393_s_at 1.53 0.0474 NAA50 N(alpha)-acetyltransferase 50, NatE catalytic subunit 1554794_a_at 1.53 0.00035 UBE3C ubiquitin protein ligase E3C 200917_s_at 1.53 0.0168 SRPR signal recognition particle receptor (docking protein) 210785_s_at 1.53 0.0463 C1orf38 open reading frame 38 203052_at 1.53 0.0404 C2 complement component 2 1555355_a_at 1.53 0.0191 ETS1 v-ets erythroblastosis virus E26 oncogene homolog 1 (avian) 210186_s_at 1.53 0.0372 FKBP1A FK506 binding protein 1A, 12kDa 211536_x_at 1.53 0.0453 MAP3K7 mitogen-activated protein kinase kinase kinase 7 1567013_at 1.53 3.81E-05 NFE2L2 nuclear factor (erythroid-derived 2)-like 2 201042_at 1.53 0.0388 TGM2 transglutaminase 2 (C polypeptide, protein-glutamine-gamma-glutamyltransferase) 237215_s_at 1.53 0.00967 TFRC transferrin receptor (p90, CD71) 1555814_a_at 1.52 0.00365 RHOA ras homolog gene family, member A 217309_s_at 1.52 0.00296 DSCR3 Down syndrome critical region gene 3 224039_at 1.52 3.97E-05 FAM186B family with sequence similarity 186, member B 1554333_at 1.52 0.00333 DNAJA4 DnaJ (Hsp40) homolog, subfamily A, member 4 206834_at 1.52 0.037 HBD hemoglobin, delta 200983_x_at 1.52 0.00159 CD59 CD59 molecule, complement regulatory protein 205787_x_at 1.52 0.0118 ZC3H11A zinc finger CCCH-type containing 11A 230397_at 1.51 0.00188 SAR1B SAR1 homolog B (S. cerevisiae) 1552928_s_at 1.51 0.0332 TAB3 TGF-beta activated kinase 1/MAP3K7 binding protein 3 209136_s_at 1.51 0.00455 USP10 ubiquitin specific peptidase 10 220580_at 1.51 0.0112 BICC1 bicaudal C homolog 1 (Drosophila) 201615_x_at 1.51 0.000258 CALD1 caldesmon 1 1555628_a_at 1.51 0.0299 HAVCR2 hepatitis A virus cellular receptor 2 223585_x_at 1.51 0.00808 KBTBD2 kelch repeat and BTB (POZ) domain containing 2 1554986_a_at 1.51 0.00237 SNX19 sorting nexin 19 1558334_a_at 1.51 4.07E-06 C22orf15 chromosome 22 open reading frame 15 230492_s_at 1.51 0.0191 GPCPD1 glycerophosphocholine phosphodiesterase GDE1 homolog (S. cerevisiae) 207334_s_at 1.51 0.00713 TGFBR2 transforming growth factor, beta receptor II (70/80kDa) 1554159_a_at 1.51 0.0136 ZMYND11 zinc finger, MYND domain containing 11 208653_s_at 1.51 0.0452 CD164 CD164 molecule, sialomucin 227621_at 1.51 0.000175 WTAP Wilms tumor 1 associated protein 203472_s_at 1.51 0.0363 SLCO2B1 solute carrier organic anion transporter family, member 2B1 1555786_s_at 1.50 0.000747 C14orf34 chromosome 14 open reading frame 34 200744_s_at 1.50 0.000213 GNB1 guanine nucleotide binding protein (G protein), beta polypeptide 1 1554834_a_at 1.50 0.00327 RASSF5 Ras association (RalGDS/AF-6) domain family member 5 204924_at 1.50 0.0146 TLR2 toll-like receptor 2 200900_s_at 1.50 0.0247 M6PR mannose-6-phosphate receptor (cation dependent) 221602_s_at 1.50 0.0239 FAIM3 Fas apoptotic inhibitory molecule 3 244563_at 1.50 0.0434 QSER1 glutamine and serine rich 1 228359_at 1.50 8.34E-05 UBASH3B ubiquitin associated and SH3 domain containing B