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Abnormal miR-148b Expression Promotes Aberrant Glycosylation of IgA1 in IgA Nephropathy

† † Grazia Serino,* Fabio Sallustio,* Sharon N. Cox,* Francesco Pesce,* and † Francesco P. Schena*

*Nephrology, Dialysis and Transplantation Unit, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy; and †Centro Addestramento Ricerca Scientifica in Oncologia (C.A.R.S.O.) Consortium, Valenzano, Italy

ABSTRACT Aberrant O-glycosylation in the hinge region of IgA1 characterizes IgA nephropathy. The mechanisms underlying this abnormal glycosylation are not well understood, but reduced expression of the enzyme core 1, b1,3-galactosyltransferase 1 (C1GALT1) may contribute. In this study, high-throughput microRNA (miRNA) profiling identified 37 miRNAs differentially expressed in PBMCs of patients with IgA nephrop- athy compared with healthy persons. Among them, we observed upregulation of miR-148b, which poten- tially targets C1GALT1. Patients with IgA nephropathy exhibited lower C1GALT1 expression, which negatively correlated with miR-148b expression. Transfection of PBMCs from healthy persons with a miR-148b mimic reduced endogenous C1GALT1 mRNA levels threefold. Conversely, loss of miR-148b function in PBMCs of patients with IgA nephropathy increased C1GALT1 mRNA and protein levels to those observed in healthy persons. Moreover, we found that upregulation of miR-148b directly correlated with levels of galactose-deficient IgA1. In vitro, we used an IgA1-producing cell line to confirm that miR-148b modulates IgA1 O-glycosylation and the levels of secreted galactose-deficient IgA1. Taken together, these data suggest a role for miRNAs in the pathogenesis of IgA nephropathy. Abnormal expression of miR-148b may explain the aberrant glycosylation of IgA1, providing a potential pharmacologic target for IgA nephropathy.

J Am Soc Nephrol 23: ccc–ccc, 2012. doi: 10.1681/ASN.2011060567

IgA nephropathy (IgAN) is considered the most IgAN is a complex multifactorial disease whose common form of primary GN throughout the pathogenic mechanism is still unknown. Several world, and about 40% of patients develop ESRD.1,2 investigators have sought to identify specific genetic The initial event in the pathogenesis of IgAN is markers associated with the development and pro- the mesangial deposition of IgA1, aberrantly gly- gression of this disease;15 however, few studies have cosylated because the hinge-region O-linked gly- specifically described intracellular mechanisms as- cans lack galactose.3–6 Different studies suggest that sociated with disease development.16 Recently, our this alteration could lead to IgA1 self-aggregation,7 group identified new mechanisms associated with IgA1-IgG immune complex formation,8,9 and com- plex defective clearance, with sequential deposition in the glomeruli.10 Received June 13, 2011. Accepted December 19, 2011. In humans, each IgA1 heavy-chain hinge region G.S. and F.S. contributed equally to this work. fi has three to ve 5 O-linked glycans that are built Published online ahead of print. Publication date available at by stepwise addition of monosaccharides, begin- www.jasn.org. ning with the addition of N-acetylgalactosamine Correspondence: Dr. Francesco P. Schena, Nephrology Dialysis by the enzyme N-acetylgalactosaminyltransferase and Transplantation Unit, Department of Emergency and Organ 2 and continuing with the addition of galactose Transplantation, University of Bari, Policlinico, Piazza G. Cesare bytheenzymecore1,b1,3-galactosyltransferase 1 no. 11, 70124, Bari, Italy. Email: [email protected] – (C1GALT1).11 14 Copyright © 2012 by the American Society of Nephrology

J Am Soc Nephrol 23: ccc–ccc, 2012 ISSN : 1046-6673/2305-ccc 1 BASIC RESEARCH www.jasn.org the pathogenesis of IgAN and showed that WNT-b-catenin and PI3K/Akt pathways were highly activated in patients with IgAN.17 The basis for the abnormal glycosylation in IgAN is still unknown, but some studies suggest that C1GALT1 could be involved because of its altered expression.18 On the other hand, the potential role of microRNAs (miRNAs) in the IgAN pathogenesis has been poorly in- vestigated. After the discovery of miRNAs, great effort has focused on determining their biologic functions and their relevance to diseases. In fact, deregulation of miRNAs has been associated with several disease states, including kidney diseases.19 To our knowledge, this study is the first to evaluate the global miRNA expression profile of IgAN patients9 PBMCs, which are directly involved in the disease.20 We defined the miRNA signature in patients with IgAN and showed that miR-148b regulating C1GALT1 explains the abnormal glycosylation pro- cess in IgAN. These results support an important and unre- ported role of this miRNA in the pathogenesis of IgAN and suggest a pharmacologic rationale for the potential use of syn- thetic miRNA inhibitors to attenuate IgA1 deglycosylation in the disease.

RESULTS Figure 1. Unsupervised hierarchical clustering of miRNA ex- pression profile. miRNA expression pattern of PBMCs of seven Identification of Differentially Expressed miRNAs in patients with IgAN and seven healthy subjects (HSs) were ex- Patients with IgAN amined using Agilent array composed of 723 human and 76 The role of miRNA expression in the pathogenesis of IgAN human viral miRNAs. A total of 147 miRNAs were expressed in all samples, discriminating patients with IgAN from HSs (P,0.0001; has not been well explored. To identify miRNAs differentially , fi fi FDR 0.01). Two principal clusters were identi ed on the basis of expressed in IgAN, we analyzed their global expression pro le differential miRNA expression. in PBMCs of seven patients with IgAN and seven healthy participants. Among 723 human miRNAs represented on the microarrays, 147 were expressed in each sample. Unsupervised hierarchical clustering analysis generated a tree with the IgAN target genes of the 35 upregulated miRNAs. To reduce the and healthy participants clearly separated into two groups number of false-positive results, we used four different (Figure 1). This separation was further confirmed by display- algorithms and listed only putative target genes predicted by ing the relationships among miRNA expression patterns using at least two of them. On the basis of the results of bioinfor- principal component analysis (Supplemental Figure 1). After matics analysis, we found that one of the potential targets of we applied a fold change threshold . 2 (false discovery rate miR-148b was the gene C1GALT1, which plays an important [FDR], 0.01), 35 miRNAs were found to be significantly up- role in the pathogenesis of IgAN. Of note, other miR-148b regulated and 2 were significantly downregulated in IgAN (Sup- putative target genes were inversin (INVS) and phosphatase plemental Table 1). To validate microarray results, we and tensin homologue (PTEN) (Supplemental Table 2), two performed quantitative real-time PCR (qRT-PCR) for miR- genes that we found downregulated in patients with IgAN.17 148b, miR-188-5p, miR-361-3p, miR-886-3p, let-7b, and let-7d Ingenuity Pathway Analysis (IPA) software was then used to on miRNAs isolated from PBMCs of an independent set of 10 evaluate the biologic interaction among miRNAs. Weuploaded patients with IgAN and 10 healthy persons with the same clini- the 35 upregulated miRNAs in IPA, and two networks were cal and demographic characteristics as those in the population identified. When we merged each of them with the network used for microarray experiments (Table 1). The expression of all resulting from the gene expression profile published in our analyzed miRNAs was significantly higher in patients with IgAN, previous work,17 we found that miRNAs were strongly inter- thereby confirming microarray results (Figure 2, A and B). connected with the mRNA network (Supplemental Figure 2). In particular, let-7d directly regulated PTEN, miR-361 regu- In silico Analysis of miRNA Targets lated INVS, miR-148b regulated both INVS and PTEN,and To study the molecular mechanisms in which the miRNAs are three miRNAs (let-7d, let-7a, miR-98) indirectly regulate AKT involved, we performed a bioinformatic analysis to predict through RET.

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Table 1. Demographic and clinical features of patients and healthy participants Initial Sample Cohort Disease Controls Validation Sample Cohort Variable Healthy Healthy IgAN MPGN-I FSGS HSPN IgAN Participants Participants Participants (n) 25 25 3 5 10 50 50 Men/women (n/n)16∕915∕10 2/1 3/2 5/5 38/12 40/10 Age (yr) 37.2610.2 36.1610.6 3569.6 42612.3 8.56336612.3 43610.6 Serum creatinine (mg/dl) 0.960.2 0.860.3 0.760.05 0.860.4 0.560.1 1.0060.5 0.960.3 Estimate GFR (ml/min per 1.73 m2)118622.3 106613.1 122610 116.8611 ND 120611 114612.3 Proteinuria (g/24 h) 0.360.1 0.160.2 2.761.2 2.0560.9 ND 0.860.2 0.0660.02 Systolic BP (mmHg) 121620.4 120610 133611.5 126615 118616 124613.3 11969 Diastolic BP (mmHg) 726974688060.5 7966.5 6768 77.269.7 7469 Total IgA (ng/ml) 0.1060.004 0.1260.01 ND ND 0.1560.01 0.1460.002 0.1360.001 Unless otherwise noted, values are expressed as mean 6 SD. IgAN, IgA nephropathy; MPGN-I, membranoproliferative GN type I; FSGS, focal segmental glomerulosclerosis; HSPN, Henoch-Schönlein purpura nephritis; ND, not determined.

miR-148b levels in patients with IgAN and the healthy participants. A negative corre- lation was observed (R2=0.4; P,0.01; Fig- ure 4C): samples with higher miR-148b expression levels showed lower levels of C1GALT1 mRNA. The inverse correlation observed be- tween the levelsofmiR-148bandC1GALT1 mRNA suggests that this gene is probably a target of miR-148b. Moreover, we performed a bioinformatic Figure 2. Differential expression validation. Validation of differential expression of miR-148b, miR-188-5p, miR-886-3p, let-7b, and let-7d (A) and miR-361-3p (B) in an analysis to estimate the effect of some single- independent set of PBMCs from 10 patients with IgAN and 10 healthy subjects (HSs). nucleotide polymorphisms (SNPs) on puta- Expression levels were quantified using qRT-PCR. The miRNA relative expressions were tive miRNA targets interrogating the 39-UTR normalized to the expression of U6. Expression levels of miR-148b, miR-188-5p, miR- and predicting whether a SNP within the 886-3p, let-7b, let-7d, and miR-361-3p were found to be significantly higher in pa- target site will disrupt/eliminate or enhance/ tients with IgAN than in HSs. The histograms represent the mean 6 SEM. *P,0.03; create a miRNA binding site. Our results re- **P,0.01. vealed that the 1365G/A polymorphism (rs1047763) in the 39-UTR of C1GALT1, which is associated with IgAN,21,22 affects Identification of miR-148b as a Regulator of C1GALT1 miR-148b binding sites in this gene. In fact, the biocomputa- Expression tional analysis showed that the 1365G allele enhances miR-148b C1GALT1 is known as a directly involved in IgAN; however, binding (Figure 4D). the basis for its decreased function in the disease is unknown. To study whether the upregulated expression of miR-148b Therefore, we evaluated the C1GALT1 mRNAexpression levels was specific to IgAN, we checked the miR-148b expression in by qRT-PCR in the same set of RNA samples used in the PBMCs from controls with three additional diseases: 3 patients microarray validation. We found that C1GALT1 levels were with membranoproliferative GN type I , 5 with focal segmental significantly lower in patients with IgAN than in the healthy glomerulosclerosis, and 10 with Henoch–Schönlein purpura participants (P,0.0001; Figure 3). nephritis. miR-148b expression in these diseases has been The in silico analysis showed that the miR-148b increase compared with that of IgAN and controls. We found that the could be the cause of the C1GALT1 reduction. In fact, se- miR-148b was again higher in patients with IgAN (P,0.0001) quence alignment of human miR-148b with 39-untranslated than in those with the other three diseases (Figure 5), con- region (UTR) C1GALT1 identified a binding site (nucleotides firming that higher miR-148b levels are typical of IgAN. 1355–1361 in human C1GALT1, chr 12q13; Figure 4A) that is well conserved among different species (Figure 4B). miR-148b Downregulates C1GALT1 mRNA Expression To study whether the downregulation of C1GALT1 in The nature of target of miR-148b and its mode of action in patients with IgAN was attributable to increased miR-148b PBMCs have not been functionally identified; thus, we tested levels, we tested the correlation between C1GALT1 mRNA and whether miR-148b was able to modulate the expression of

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C1GALT1. For this purpose, we performed some transient mimic, showed a three-fold reduction in endogenous C1GALT1 transfection experiments ex vivo using PBMCs from an inde- mRNA levels (P,0.03; Figure 6A). pendent group of four patients with IgAN and four healthy To provide evidence for a direct effect of miR-148b on persons. C1GALT1 expression, we transfected PBMCs isolated from five We increased the amount of the endogenous miR-148b other healthy persons with both a miR-148b mimic and a within PBMCs isolated from four healthy persons, transfecting mutated miR-148b mimic (the D20-21 miR-148b, with a short RNA sequences that mimic the action of the miRNA, to substitution from UG to AC in position 20-21 [59→39]). simulate the situation found in patients with IgAN. PBMCs of We found that the C1GALT1 mRNA expression decreased the four healthy persons, transfected with 25nM miR-148b only after transfection with the wild-type miR-148b mimic (Figure 6B). To further confirm that C1GALT1 mRNA was targeted by miR-148b, we performed the reverse experiment, in which we used synthetic hairpin miRNA inhibitors to silence the activity of miR-148b in PBMCs isolated from four patients with IgAN. The miRNA inhibitors were chemically modified acids de- signed to bind and to inhibit endogenous miR-148b. The transient transfection of IgAN PBMCs with 250 nM of miR- 148b inhibitor led to a three-fold increase in endogenous C1GALT1 mRNA levels (P,0.01, Figure 6C). In line with our computational analysis, we showed that miR-148b did not affect Cosmc chaperone, which is involved in stability and activity of C1GALT123 (Supplemental Figure 3).

miR-148b Downregulates C1GALT1 Protein Expression Figure 3. C1GALT1 gene expression levels evaluated by real- time PCR in 10 patients with IgAN and 10 healthy subjects (HSs). We used the same strategy to functionally inhibit or enhance C1GALT1 expression levels were significantly lower in patients the mature form of miR-148b in PBMCs of patients with IgAN with IgAN than in HSs. C1GALT1 expression levels were nor- and healthy persons, respectively, with the aim of establish- malized on the housekeeping gene b-actin. The scatter plots ing whether C1GALT1 protein expression was also effec- represent the mean 6 SEM. #P,0.0001. tively controlled by miR-148b. PBMCs of five healthy persons

Figure 4. miR-148b targets C1GALT1. (A) Genomic localization of miR-148b in chromosome 12q13. (B) Sequence alignment of the miR- 148b base-pairing sites in the 39-UTR of C1GALT1 mRNA showing that the regions complementary to miR-148b are highly conserved among human, chimp, mouse, rat, dog, and chicken. The “seed” sequences of miR-148b complementary to C1GALT1 are shown in gray. (C) Linear correlation between the expression of C1GALT1 and the expression of miR-148b in PBMCs of 10 patients with IgAN and 10 healthy subjects (HSs). C1GALT1 mRNA levels inversely correlated with miR-148b expression levels (R2=0.4; P,0.01). (D) Output of bioinformatic analysis performed by MicroSNiPer algorithms showing that the 1365G/A polymorphism (rs1047763) affects miR-148b binding sites in the 39-UTR of C1GALT1. The 1365G allele (arrow) enhances miR-148b binding, whereas the 1365A allele does not appear in the output as a possible binding site for miR-148b.

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transiently transfected with 25 nM of miR-148b mimic resulted in a remarkable reduction in C1GALT1protein expression (2.2- fold; P,0.002) as revealed by Western blot. Of note, the in- crease in the miR-148b level in PBMCs of healthy persons resulted in the same C1GALT1 protein expression levels as in PBMCs of patients with IgAN (Figure 7A). On the contrary, in the reverse experiment, PBMCs of five patients with IgAN transfected with 250 nM of miR-148b inhibitor showed a sig- nificant increase in C1GALT1 protein production (1.7-fold in- crease; P,0.01). Surprisingly, C1GALT1 protein levels in patients with IgAN increased to the same levels as in healthy persons (Figure 7B). Altogether our results demonstrate that Figure 5. miR-148b expression levels evaluated by real-time PCR miR-148b targets C1GALT1, which is responsible for a de- in PBMCs from 3 patients with membranoproliferative GN type I creased activity of the gene in patients with IgAN. (MPGN-I), 5 with focal segmental glomerulosclerosis (FSGS), and 10 with Henoch-Schönlein purpura nephritis (HSPN). miR-148b miR-148b Expression and Altered IgA1 Glycosylation expression in these diseases and in healthy subjects (HSs) has Relationship been compared with that in IgAN. We found that miR-148b levels To further validate the relationship between miR-148b ex- were higher in patients with IgAN than in those with MPGN-I, pression and altered IgA1 glycosylation, we enrolled an in- FSGS, and HSPN and in HSs. The miRNA relative expressions dependent cohort of 50 patients with IgAN and 50 healthy were normalized to the expression of U6. The histograms repre- persons. Again, miR-148b expression levels, evaluated by 6 , # , sent the mean SEM. **P 0.01; P 0.0001. qRT-PCR, were significantly higher in PBMCs of patients with IgAN than in those of healthy persons (P,0.001; Figure 8A). We next evaluated the galactose-deficient (Gal-deficient) IgA1 in serum of the same patients with IgAN and healthy persons using helix aspersa agglutinin (HAA) lec- tin binding assay. We found that the Gal- deficient IgA1 was significantly higher in patients with IgAN (mean OD 6 SD, 0.9860.1) than in healthy persons (mean OD, 0.5360.02; P,0.0001) (Figure 8B), whereas total IgA levels were similar in the two groups (0.13760.002 and 0.1326 0.002 mg/ml, respectively) (Figure 8C). Pear- son correlation analysis between miR-148b expression level and Gal-deficient IgA1 level showed a significant positive correla- tion (r =0.4;P,0.0001; Figure 8D), sup- porting our data on C1GALT1 regulation Figure 6. miR-148b regulates C1GALT1 mRNA expression. (A) C1GALT1 expression by miR-148b and explaining the abnormal levels were analyzed by real-time PCR in PBMCs of healthy subjects (HSs) after increase in Gal-deficient IgA1 consequent transfection with miR-148b mimic. Increasing the amount of miR-148b within the to higher miR-148b levels in patients with PBMCs of HSs resulted in a three-fold reduction in endogenous C1GALT1 mRNA IgAN. To confirm the direct cause-and- levels. (B) C1GALT1 expression levels were analyzed by real-time PCR in PBMCs of five effect relationship between miR-148b ex- other HSs after transfection with the mutated miR-148b mimic, D20-21, and miR-148b pression and Gal-deficient IgA production, mimic wild-type. C1GALT1 mRNA expression decreased only after transfection with we used as a model the human B lymphoma the wild-type miR-148b mimic. (C) C1GALT1 expression levels were analyzed by real- DAKIKI cells producing IgA1.24 We trans- time PCR in PBMCs of patients with IgAN after transfection with miR-148b inhibitor. fected DAKIKI cells with miR-148b mimic Silencing the activity of miR-148b within IgAN PBMCs led to a three-fold increase in endogenous C1GALT1 mRNA levels. “Mock” indicates mock-transfected cells going and inhibitor and evaluated changes in the fi through the transfection processes without addition of mimic/inhibitor miRNA. Expres- Gal-de cient IgA1 levels by means of HAA sion data were normalized on the housekeeping gene b-actin. Data are representative lectin binding assay. DAKIKI cells transfec- of four independent experiments for A and C and five independent experiments for B ted with 50 nM of miR-148b mimic resulted (mean 6 SEM). **P,0.01. in a remarkable increase in Gal-deficient

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C1GALT1 are again unknown. Accumulat- ing evidence suggests the involvement of miRNAs in the pathogenesis of some kid- ney diseases, such as acute kidney rejection, lupus nephritis, and renal ischemia reper- fusion injury.28–30 However, miRNAs po- tentially involved in the pathogenesis of IgAN have been partially studied in renal tissue.31 Starting from a microarray miRNA ex- pression profile in PBMCs of patients with IgAN and using several bioinformatic ap- proaches, we identified 37 miRNAs that are differentially regulated in IgAN patients compared with healthy persons, along with Figure 7. miR-148b regulates C1GALT1 expression at protein level. (A) Transfection of their candidate target genes. PBMCs of healthy participant with 25 nM miR-148b mimic resulted in a 2.2-fold re- Furthermore, we overlapped modulated duction in C1GALT1 protein expression. Increasing the miR-148b levels within PBMCs miRNAs with previously identified gene of healthy participants resulted in the same C1GALT1 protein levels as in IgAN PBMCs, pathways involved in IgAN17 to identify as shown by Western blot. (B) Western blot representing protein levels of C1GALT1 in candidate target genes involved in the path- fi IgAN PBMCs after transfection with 250 nM miR-148b inhibitor. A signi cant increase in ogenesis. This analysis revealed that the C1GALT1 protein production was shown in IgAN PBMCs transfected with miR-148b miRNAs and genes differentially expressed inhibitor (1.7-fold increase). miR-148b loss of function led to the same C1GALT1 in IgAN were strongly interconnected, par- protein levels as in PBMCs of healthy participants. “Mock” indicates mock-transfected cells going through the transfection processes without addition of mimic/inhibitor ticularly the six miRNAs whose expression miRNA. In both experiments, b-actin was used as endogenous control. Data are we validated by qRT-PCR. representative of five independent experiments (mean 6 SEM). **P,0.01, §P# 0.001. Among 37 miRNAs significantly dereg- ulated in patients with IgAN, miR-148b IgA1 levels (1.5-fold increase; P,0.002; Figure 8E). In con- aroused our interest because its putative target genes were trast, in the reverse experiment, DAKIKI cells transfected with C1GALT1, INVS,andPTEN, three genes downregulated in 500 nM of miR-148b inhibitor showed a significant reduction these patients.11,17 Wefoundareducedexpressionof in Gal-deficient IgA1 levels (1.8-fold; P,0.01; Figure 8E). C1GALT1 in patients with IgAN, and we showed that miR- These data further confirmed the role of miR-148b in IgA1 148b expression negatively correlated with the C1GALT1 ex- O-glycosylation regulation. pression levels in the same patients. In the last years, two large-scale case-control association studies pointed out the role of C1GALT1 in the pathogenesis DISCUSSION of IgAN, reporting some C1GALT1 genetic variants associated with the genetic susceptibility to IgAN in the Italian and IgAN is the most common primary form of GN worldwide,25 Chinese populations.21,22 Both studies identified a positive leading to ESRD in 40% of patients. There is a consensus association between the susceptibility to IgAN and a opinion that aberrant glycosylation of IgA1 is directly involved C1GALT1 SNP at the position 1365 G/A (rs1047763) in the in the pathogenesis of the disease.3–5,10,26 This abnormality is 39-UTR regulatory region. Furthermore, a correlation be- manifested by a deficiency of galactose in the hinge-region O- tween this SNP and a reduced C1GALT1 expression in homo- linked glycans of IgA1. Biosynthesis of these glycans begins zygous 1365G/G individuals was shown.22 with the addition of N-acetylgalactosamine by the enzyme N- Our bioinformatic analysis revealed that rs1047763 SNP acetylgalactosaminyltransferase 2 and continues with the ad- is located within the miR-148b binding site on the C1GALT1 dition of galactose by the enzyme C1GALT1.11–14 39-UTR region, and the binding affinity of miR-148b is greater The mechanisms leading to this aberrant glycosylation have when the 1365G-variant allele is present. These results could been studied extensively; however, to date, the nature of explain the lower C1GALT1 expression levels in patients with abnormally O-glycosylated IgA1 is still obscure. Kudo et al. IgAN carrying the 1365G/G genotype.22 demonstrated an important role for C1GALT1 in the molec- Supporting our bioinformatic analysis, we found that ular basis for the aberrant IgA1 glycosylation in IgAN.18 How- C1GALT1 mRNA levels were significantly lower in patients ever, it is controversial whether IgA1 is undergalactosylated with IgAN. Moreover, using the strategy to functionally en- as a consequence of functional changes in C1GALT1 activity or hance or inhibit the mature form of miR-148b, we confirmed as a consequence of its reduced expression.14,27 Moreover, the biologically, in an ex vivo experiment, that miR-148b targets molecular mechanisms behind the impaired functioning of C1GALT1 and can modulate its mRNA and protein levels. Of

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persons were associated with low miR-148b levels. This finding supports our data on C1GALT1 regulation by miR-148b and explains the abnormal increase in Gal- deficient IgA1 due to high miR-148b levels in patients with IgAN. The role of miR-148b in IgA1 O-glycosylation regulation was also confirmed by transfection experiments in DAKIKI human B lymphoma cells showing that miR-148b modulated levels of secreted Gal-deficient IgA1. On the basis of our results, we can hypothesizethatinIgAN,lowerlevelsof C1GALT1 due to the inhibiting effect of overexpressed miR-148b may lead to an increase in circulating deglycosylated IgA1.8,9,32–34 These galactose-deficient IgA1 molecules are predisposed to aggre- gation and formation of immune com- plexes;8 they are recognized by naturally occurring circulating antibodies.9 Conse- quently, these complexes are less effectively cleared from the circulation35 and effi- ciently bind to mesangial cells, inducing cellular proliferation, overproduction of Figure 8. miR-148b regulates Gal-deficient IgA1 levels. (A) The miR-148b expression extracellular matrix, and synthesis of in- levels evaluated by real-time PCR in 50 patients with IgAN and 50 healthy subjects flammatory cytokines that can initiate or (HSs). miR-148b expression levels were significantly higher in patients with IgAN than in perpetuate the course of GN.36–38 HSs. miR-148b expression levels were normalized to the expression of U6. The his- Our study has one important limitation. tograms represent the mean 6 SEM. §P,0.001. (B) shows the serum levels of Gal- We did not study the effect of miR-148b deficient IgA1 in patients with IgAN and HSs. The Gal-deficient IgA1 was significantly on the process of IgA1 glycosylation in an higher in sera obtained from the patients with IgAN than in sera from HSs. The relative experimental model of IgAN because IgA1 lectin binding per unit IgA1 was calculated as the OD value of lectin over the OD value is present exclusively in humans and hom- of total IgA. (C) The serum level of total IgA in patients with IgAN and HSs determined inoid primates.39,40 However, our study by ELISA. IgA levels in sera obtained from 50 patients with IgAN and 50 HSs were similar. The histograms represent the mean 6SEM. #P,0.0001. (D) Linear correlation points overall to a new regulatory mecha- between the expression of miR-148b and the Gal-deficient IgA1 levels in 50 patients nism of IgAN that can explain the aberrant with IgAN and 50 HSs. miR-148b levels directly correlated with Gal-deficient IgA1 glycosylation of IgA1 responsible for the levels (r=0.4,; P,0.0001). (E) The Gal-deficient IgA1 levels in supernatants obtained pathogenesis of the disease. Moreover, the from DAKIKI cells after transfection with miR-148b mimic and inhibitor. A significant miR-148b expression levels may be used to increase in Gal-deficient IgA1 production was shown in DAKIKI cells transfected with develop a method to diagnose the disease 50 nM miR-148b mimic (1.5-fold increase). On the contrary, the transfection with 500 through a noninvasive technique, which nM miR-148b inhibitor led to a significant reduction in Gal-deficient IgA1 levels (1.8- may replace, in the future, the renal biopsy fold). The relative lectin binding per unit IgA1 was calculated as the OD value of lectin as a diagnostic approach. 6 § , over the OD value of total IgA. The histograms represent the mean SEM. P 0.001. In conclusion, our study shows, for the first time, a global miRNA expression pro- file in PBMCs of patients with IgAN and a note, the loss of function of miR-148b in PBMCs isolated from miRNA signature. We also provide evidence for a previously patients with IgAN led to C1GALT1 protein levels similar to unknown key regulatory mechanism of IgAN by discover- those observed generally in healthy persons. In contrast, miR- ing the deregulation of miR-148b, which could explain the 148b overexpression in PBMCs isolated from healthy persons aberrant glycosylation of IgA1 in the disease. These findings led to a reduction in C1GALT1 protein to levels similar to those could also have therapeutic implications because the inhibition expressed in patients with IgAN. of miR-148b expression reverses the lower levels of C1GALT1 Moreover, patients with IgAN who had higher serum levels typical of IgAN. Therefore, miR-148b levels may be manip- of Gal-deficient IgA1 also showed higher levels of miR-148b. In ulated to provide useful new therapeutic approaches to the contrast, lower levels of serum Gal-deficient IgA1 in healthy disease.

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CONCISE METHODS Total RNA, including small RNA fractions, was then eluted in ribonuclease-free water. The RNAconcentrationwas determined with Sample Collection NanoDrop Spectrophotometer (Nanodrop Technologies). The A total of 75 patients with IgAN, 75 healthy participants, 10 patients miRNA quality was assessed using Agilent 2100 Bioanalyzer (Agilent with Henoch-Schönlein purpura nephritis, 3 patients with mem- Technologies), and only samples with RNA integrity number . 8.5 branoproliferative GN type I, and 5 patients with focal segmental were used. glomerulosclerosis were included in this study. Written informed consent was obtained from each patient and healthy participant. miRNA Microarray The study was initially conducted on a cohort of 25 patients with miRNA microarray analysis was performed using the Agilent Human IgAN and 25 healthy participants. Seven in each group were included miRNA Microarrays V2, which were based on Sanger miRBase release in the microarray experiment, 10 participants from each group were 10.1, according to the manufacturer’s protocol. Briefly, 600 ng of used for microarray validation, and 8 participants from each group total RNA isolated from PBMCs of seven patients with IgAN and were used for the in vitro transfections. Patients with IgAN had nor- seven healthy participants were first dephosphorylated with a calf in- mal renal function; patients with severe renal damage were excluded testine alkaline phosphatase treatment for 30 minutes at 37°C before from the study. Each patient was selected according to the following labeling. Samples were diluted with DMSO, denatured for 10 minutes – demographic and clinical features: age (18 50 years), moderate histo- at 100°C, and labeled using pCp-Cy3 in T4 RNA ligation buffer. fi 41 logic lesions (G2 according to the Schena classi cation ), creatinine The labeled RNA was hybridized, washed, stained, and scanned . 2 clearance 90 ml/min per 1.73 m (evaluated by the Cockcroft-Gault with an Agilent microarray scanner (G2565BA, Agilent). Microarray formula), and clinical follow-up (at least 7 years). The healthy partic- data analysis was performed using Agilent Feature Extraction Software ipants were selected on the basis of their demographic characteristics 9.5.1.1. and overlapped completely with IgAN group. All healthy participants Microarray data are available under accession number GSE25590 had a negative result on a urine test for blood and proteins. All patients at the Gene Expression Omnibus (http://www.ncbi.nlm.nih.gov/geo/). with diabetes, chronic lung diseases, neoplasm, or inflammatory dis- eases and patients receiving antibiotics, corticosteroids, and nonste- Statistical Analyses and Bioinformatics roidal anti-inflammatory agents were excluded from the study. No For microarray analysis, the raw expression signals were log- patients had symptomatic coronary artery diseases or a family his- transformed, normalized, and filtered according to the median cor- tory of premature cardiovascular diseases. rected signal of all the miRNAs with an intensity .100 (which is To validate some of the results obtained, we collected blood considered as expressed), which resulted in the selection of 147 samples from an independent cohort of 50 patients with IgAN and 50 miRNAs out of the original set of 723 miRNAs. The preprocessed healthy persons matched in their demographic and clinical features microarray data were imported into the R language for statistical with the participants included in the first group. analysis computing (http://www.r-project.org). miRNAs displaying Ten patients with Henoch-Schönlein purpura nephritis, three differential expression between patients with IgAN and healthy par- with membranoproliferative GN type I, and five with focal segmental ticipants were detected using a two-sample t test. Probe sets were glomerulosclerosis were selected as disease controls. sorted after a P value showed a statistically significant difference All participants were from the Italian population. The study was and were adjusted to account for multiple testing using the FDR carried out according to the principles of the Declaration of Helsinki method of Storey and Tibshirani.42 Only genes that were significantly and was approved by our institutional ethics review board. (FDR,0.01) modulated in patients with IgAN compared with healthy The main demographic and clinical features of our patients and participants were considered for further analysis. Significance of differ- controls included in the study are summarized in Table 1. There were ential expression, as determined by the enrichment analysis, was per- no statistically significant differences between the IgAN and healthy mutated 1000 times. To determine the features that are most closely groups for any measure considered. correlated with IgAN phenotype, we applied a most stringent filter with PBMCs were isolated by density separation over a Ficoll-Hypaque FDR,0.01 and a fold change . 2. Two-dimensional hierarchical clus- (Ficoll-Paque Plus, GE Healthcare) gradient (460 g for 30 minutes). tering was performed using Spotfire decision site 8.0 (http://spotfire. PBMCs were washed three times with PBS (pH 7.4)/1 mM EDTA tibco.com). To assess biologic relationships among genes, we used In- (Sigma). Cells were then counted, and their viability was determined genuity Pathway Analysis software (IPA, Ingenuity System, Redwood by trypan blue exclusion. City, CA). IPA computes a score for each network according to the fitof Serum was collected at the same clinical time point, and whole the set of supplied focus miRNAs/genes. These scores indicate the likeli- blood was centrifuged at 3000 rpm for 10 minutes at room tem- hood that focus genes would belong to a network versus those ob- perature. tained by chance. miRNA targets were predicted by means of miR- Base 13.0 (http://microrna.sanger.ac.uk),43 TargetScan 5.1 (http:// miRNA Isolation www.targetscan.org),44 PicTar (http://pictar.org),45 and miRWalk Isolated PBMCs were used for total RNA extraction by means of (http://www.ma.uni-heidelberg.de) algorithms. Potential targets were miRNeasy Mini kit (Qiagen) according to the manufacturer’s pro- chosen overlapping results from the four algorithms and selecting tar- tocol. Deoxyribonuclease treatment was carried out to remove any gets of genes predicted by at least two of the algorithms; the targets were contaminating DNA (RNase-Free DNase Set, Qiagen). selected by a score cutoff computed by a weighted sum of a number of

8 Journal of the American Society of Nephrology J Am Soc Nephrol 23: ccc–ccc,2012 www.jasn.org BASIC RESEARCH sequence and context features of the predicted miRNA:mRNA duplex. validated nonsilencing siRNA sequence AllStars Negative Control Bioinformatic analysis to estimate the effect of SNPs on putative siRNA (50 nM, Qiagen) was used as negative control and Syn-hsa- miRNA targets was carried out by microSNiPer software (Clinical miR-1 miScript miRNA mimic (25 nM, Qiagen) and anti–hsa-miR-1 Brain Disorders Branch, National Institutes of Health, Bethesda, miScript miRNA inhibitor (250 nM, Qiagen) were used as positive MD; http://cbdb.nimh.nih.gov/microsniper)46 interrogating the controls for miR-148b mimic and miR-148b inhibitor transfection, 39-UTR and predicting whether a SNP within the target site will respectively. Each transfection experiment was done in triplicate. disrupt/eliminate or enhance/create a microRNA binding site. A After transfection, cells were incubated for 24 hours or 48 hours at banded Smith-Waterman dynamic program was used along the 37°C in 5% CO2 and used, respectively, for total RNA extraction and best-matched regions. The algorithm output only those miRNAs/ protein extraction. seeds in which the alignment was shifted because of a change in For analysis of IgA1 glycosylation, the surface IgA1-positive the alleles. human B lymphoma cell line, DAKIKI, was purchased from A two-tailed t test was used to assess differences in biologic fea- ATCC.24,47 Cells were cultured at 1 3 105 cells per well in a 12-well tures between patients with IgAN and healthy participants. A Pearson plate with RPMI 1640 medium containing 10% heat-inactivated FBS, correlation test was used to study continuous variables. All values 1 mM sodium pyruvate, and 2 mM glutamine. The transfection of were expressed as the mean 6 SEM of data obtained from at least miRNAs mimic and inhibitor was carried out using TransIT-TKO three independent experiments. Results were considered statistically Transfection Reagent (Mirus) in accordance with the manufacturer’s significant at P,0.05. procedure. For each transfection, 50 nM of miR-148b mimic (Qiagen) and 500 nM of miR-148b inhibitor (Qiagen) were used. Each trans- qRT-PCR fection experiment was done in triplicate. After transfection, cells Total RNA, including small RNA fractions, was reverse transcribed were incubated for 72 hours at 37°C in 5% CO2. with miScript Reverse Transcription Kit (Qiagen) following the manufacturer’s instructions. Western Blot Analysis Real-time RT-PCR for the quantification of a subset of miRNAs The amount of C1GALT1, after transfection, was determined by (miR-148b, miR-188-5p, let-7d, let-7b, miR-361-3p, and miR-886-3p, Western blotting analysis. Total protein extract were prepared plus an endogenous control) was carried out with miScript Primer withlysisbuffercontaining150mMNaCl,50mMTris-HCl(pH 8), Assays and miScript SYBR Green PCR Kit from Qiagen. Real-time PCR 1% NonidetP-40, 0.1% sodium deoxycholate, and 0.1% SDS, plus amplification reactions were performed in triplicate in 25 mloffinal proteinase inhibitors. The protein concentration was determined volume via SYBR Green chemistry on iCycler (Bio-Rad). by the Bradford assay (BioRad). Eighty micrograms of each protein Normalization was performed with a small nucleolar RNA U6 lysate was separated on a 10% SDS-PAGE and transferred to endogenous control. Comparative real-time PCR was performed in polyvinylidene difluoride membrane (Millipore). The membranes triplicate, including no-template controls. Relative expression was were incubated in 5% nonfat milk powder diluted in PBS calculated using the 2-DCt method. containing 0.1% Tween-20 for 2 hours at room temperature and For C1GALT1 and Cosmc expression analysis, total RNA was probed with a mouse monoclonal anti-C1GALT1 antibody (Santa reverse-transcribed with QuantiTect Reverse Transcription Kit Cruz Biotechnology) in blocking buffer overnight at 4°C. Finally, (Qiagen) following the manufacturer’s instructions. qRT-PCR membranes were incubated with secondary antibody of horserad- amplification reactions were performed in triplicate in 25-ml final ish peroxidase conjugated goat anti-mouse IgG (BioRad). Immu- volumes using SYBR Green chemistry on an iCycler. qRT-PCR was nocomplexes were detected with enhanced chemiluminescence performed using the QuantiTect Primer Assay and the QuantiFast method (GE Healthcare). The same membranes were stripped SYBR Green PCR mix (Qiagen). Genes were amplified according to andreprobedwithanti–b-actin monoclonal antibody (Sigma). the manufacturer’sdirections.Theb-actin gene amplification was Images of autoradiography were acquired using a scanner EPSON used as a reference standard to normalize the target signal. Perfection 2580 Photo (EPSON) and quantified by Image J 1.34 Software (http://rsb.info.nih.gov/ij). The ratio between intensities of miR-148b Mimic and Inhibitor Transfection C1GALT1 and b-actin bands was used to normalize C1GALT1 ex- 6 Isolated PBMCs were cultured at 2 3 10 cells per well in a 12-well pression in each sample. plate with RPMI-1640 supplemented with 2 mM L-glutamine, 1 mM sodium pyruvate, 1 mM nonessential amino acids, 25 mM HEPES Determination of Serum IgA buffer, and 10% heat-inactivated FBS. The transfection of miRNAs IgAcontent in serum from each participant was measured in duplicate mimic and inhibitor was carried out using TransIT-TKO Transfection using ELISA. Briefly, high-adsorption polystyrene 96-microwell

Reagent (Mirus) in accordance with manufacturer’s procedure. For plates (Corning Inc.) were coated overnight with 5 mgofF(ab9)2 each transfection, 25 nM of miR-148b mimic (Qiagen), 250 nM of fragment goat anti-human IgA antibody per ml (Jackson Immuno- miR-148b inhibitor (Qiagen), and 25 nM of a mutated miR-148b Research Laboratories) in PBS at 4°C. Plates were blocked with 1% mimic (D20-21 miR-148b, with a substitution from UG to AC in BSA in PBS containing 0.05% Tween-20 for 90 minutes at room position 20-21 [59→39]; Qiagen) were used. In transfection temperature. Samples diluted in blocking buffer and standard human experiments, a mock-transfection control was performed by putting IgA (Calbiochem) were added to each well and then incubated for 90 cells through the transfection procedure without adding miRNA. The minutes at room temperature. The captured IgA was then detected

J Am Soc Nephrol 23: ccc–ccc, 2012 miR-148b in IgA Nephropathy 9 BASIC RESEARCH www.jasn.org

with biotin-labeled F(ab9)2 fragment of goat IgG anti-human IgA 2. Schena FP, Coppo R: IgA nephropathies. In: Oxford Textbook of (Biosource). The binding was measured after addition of avidin– Clinical Nephrology, 3rd ed., edited by Davison AM, Cameron JS, horseradish peroxidase conjugate (ExtrAvidin; Sigma-Aldrich), and Grünfeld JP, Ponticelli C, Ritz E, Winearls CG, van T Persele C, Oxford, Oxford University Press, 2005, pp 469–501 the reaction was developed with the peroxidase chromogenic sub- 3. Conley ME, Cooper MD, Michael AF: Selective deposition of immu- – strate o-phenylenediamine H2O2 (Sigma-Aldrich). The color re- noglobulin A1 in immunoglobulin A nephropathy, anaphylactoid pur- action was stopped with 2 N H2SO4, and the OD at 490 nm was pura nephritis, and systemic lupus erythematosus. J Clin Invest 66: determined in a microplate reader (GDV, Italy; model DV 990 B/V6). 1432–1436, 1980 – IgA1 concentration in unknown samples was determined by interpo- 4. Barratt J, Feehally J: IgA nephropathy. J Am Soc Nephrol 16: 2088 2097, 2005 lation of the respective OD into the appropriate standard curve. 5. Coppo R, Amore A: Aberrant glycosylation in IgA nephropathy (IgAN). Kidney Int 65: 1544–1547, 2004 Detection of Gal-deficient IgA1 6. Novak J, Julian BA, Tomana M, Mesteck J: Progress in molecular and Gal-deficient IgA1 was detected by binding of the lectin, HAA (Sigma- genetic studies of IgA nephropathy. JClinImmunol21: 310–327, 2001 Aldrich), which is specific for terminal GalNAc, as reported else- 7. Kokubo T, Hiki Y, Iwase H, Tanaka A, Toma K, Hotta K, Kobayashi Y: where.48,49 High-adsorption polystyrene 96-microwell plates (Corning Protective role of IgA1 glycans against IgA1 self-aggregation and adhe- sion to extracellular matrix proteins. J Am Soc Nephrol 9: 2048–2054, Inc.) were coated overnight with 3 mgofF(ab9)2 fragment goat anti- 1998 human IgA antibody per ml (Jackson ImmunoResearch Laboratories) 8. Tomana M, Matousovic K, Julian BA, Radl J, Konecny K, Mestecky J: in PBS at 4°C. Plates were blocked for 3 hours at room temperature with Galactose-deficient IgA1 in sera of IgA nephropathy patients is present 1% BSA/PBS containing 0.1% Tween-20. Samples diluted in blocking in complexes with IgG. Kidney Int 52: 509–516, 1997 buffer were added to each well and incubated overnight at 4°C. The 9. Tomana M, Novak J, Julian BA, Matousovic K, Konecny K, Mestecky J: captured IgA was subsequently desialylated by treatment for 3 hours at Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies. JClinIn- 37°C with 20 mU/ml neuraminidase from Vibrio cholerae (Sigma- vest 104: 73–81, 1999 7 Aldrich) in 10 mM sodium acetate buffer (pH 5). Samples were then 10. 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J Am Soc Nephrol 23: ccc–ccc, 2012 miR-148b in IgA Nephropathy 11 SUPPLEMENTARY MATERIAL

SUPPLEMENTARY FIGURES

Supplementary Figure 1. Principal component analysis 3D diagram showing a spatial representation of the differential miRNAs expression. IgAN patients (I) and healthy subjects (HS) were two distinct populations regarding miRNA expression levels.

Supplementary Figure 2. Functional analysis of the top modulated miRNAs identified by microarray.

The networks were algorithmically constructed by Ingenuity Pathway Analysis (IPA) software on the basis of the functional and biological connectivity of miRNAs and genes. The network is graphically represented as nodes (miRNAs or genes) and edges (the biological relationship between miRNAs/genes). Analyzing the 37 miRNAs modulated in IgAN and merging each of them with the the WNT–βcatenin and PI3K/Akt gene expression pathways previously identified, two principal networks were built. miRNAs were strongly interconnected with the mRNA networks. In particular, (A) let7d regulated PTEN, miR148b regulated both inversin (INVS) and PTEN, and let7d, let7a, miR98 indirectly regulate AKT through RET gene. (B) miR361 regulated INVS and mir17 regulated casein kinase 1alpha 1 (CSNK1A1) through karyopherin alpha 2 (KPNA2). Red and green shaded nodes represent up and downregulated genes, respectively; empty nodes represent modulated miRNAs or genes that IPA automatically includes because they are biologically linked to our genes based on the evidence in the literature.

Supplementary Figure 3. miR148b didn’t regulate Cosmc mRNA Expression. Cosmc expression levels were analyzed by realtime PCR in PBMCs of 4 healthy subjects (HS) following transfection with miR148b mimic (A) and in PBMCs of 4 IgAN patients following transfection with miR148b inhibitor (B). In both experiments, Cosmc expression remain unvaried after transfection. Mock indicates mocktransfected cells going through the transfection processes without addition of mimic/inhibitor miRNA. Expression data were normalized on the housekeeping gene βactin. Data are representative of four independent experiments (means ± SEM).

SUPPLEMENTARY TABLES

microRNA Location FDR Fold-change p-value hsamiR148b 12q13.13 0,0037 2,2913 0.0020 hsamiR1885p Xp11.23 0,0065 3,9169 0.0040 hsamiR3613p Xq21.1 0.0037 2.6396 0.0020 hsamiR8863p 5q31.2 0.0037 3.3224 0.0020 hsalet7a 9q22.32 0.0037 2.1588 0.0020 hsalet7b 22q13.31 0.0037 2.7664 0.0020 hsalet7c 21q21.1 0.0037 2.8403 0.0020 hsalet7d 9q22.32 0.0037 2.8125 0.0020 hsalet7f 9q22.32 0.0037 2.3714 0.0020 hsalet7i 12q14.1 0.0037 2.5724 0.0020 hsamiR18a 13q31.3 0.0037 2.3013 0.0020 hsamiR18b Xq26.2 0.0037 2.2687 0.0020 hsamiR98 Xp11.22 0.0037 2.5817 0.0020 hsamiR374b Xq13.2 0.0037 2.5670 0.0020 hsamiR4235p 17q11.2 0.0037 2.6512 0.0020 hsamiR128 2q21.3 0.0037 2.6402 0.0020 hsamiR134 14q32.31 0.0037 2.8265 0.0020 hsamiR7685p 16q22.3 0.0089 2.7953 0.0060 hsamiR185 22q11.21 0.0037 2.1345 0.0020 hsamiR660 Xp11.23 0.0037 2.0519 0.0020 hsamiR107 10q23.31 0.0037 2.2203 0.0020 hsamiR7 9q21.32 0.0037 2.2692 0.0020 hsamiR71* 9q21.32 0.0037 2.0477 0.0020 hsamiR425 3p21.31 0.0037 2.0149 0.0020 hsamiR500* Xp11.23 0.0037 2.2459 0.0020 hsamiR17 13q31.3 0.0037 2.1187 0.0020 hsamiR103 5q34 0.0037 2.1755 0.0020 hsamiR5023p Xp11.23 0.0037 2.2209 0.0020 hsamiR93 7q22.1 0.0037 2.3202 0.0020 hsamiR221 Xp11.3 0.0037 2.0627 0.0020 hsamiR20b Xq26.2 0.0037 2.0355 0.0020 hsamiR923 17q22 0.0037 2.9392 0.0020 hsamiR340 5q35.3 0.0065 2.1176 0.0040 hsamiR199b3p 9q34.11 0.0089 2.1679 0.0060 hsamiR3425p 14q32.2 0.0065 2.2112 0.0040 hsamiR575 4q21.22 0.0089 () 2.0807 0.0080 hsamiR451 17q11.2 0.0047 () 2.6562 0.0080

Supplementary Table 1. 37 miRNAs differently expressed in 7 IgAN patients compared to 7 healthy subjects (FDR ≤ 0.01) using microarray technology.

Suppl Table 2. miR-148b predicted target

MicroRNA Gene RefseqID Start Sequence End p-value hsa-miR-148b ZDHHC17 NM_015336 3219 UCAGUGCAUCACA 3207 0.0000 hsa-miR-148b TMED7 NM_181836 2016 UCAGUGCAUCACA 2004 0.0000 hsa-miR-148b CTSA NM_001127695 2182 UCAGUGCAUCA 2172 0.0001 hsa-miR-148b CTSL1 NM_001912 1672 UCAGUGCAUCA 1662 0.0001 hsa-miR-148b PRKCZ NM_002744 2262 UCAGUGCAUC 2253 0.0004 hsa-miR-148b DMXL1 NM_005509 9412 UCAGUGCAUCA 9402 0.0005 hsa-miR-148b KCNN3 NM_002249 2845 UCAGUGCAUC 2836 0.0005 hsa-miR-148b LAMB2 NM_002292 5760 UCAGUGCAU 5752 0.0005 hsa-miR-148b ULBP1 NM_025218 1232 UCAGUGCAUCA 1222 0.0006 hsa-miR-148b C9orf61 NM_004816 1980 UCAGUGCAUC 1971 0.0006 hsa-miR-148b SCAP NM_012235 4155 UCAGUGCAU 4147 0.0006 hsa-miR-148b PFTK1 NM_012395 4727 UCAGUGCAUCA 4717 0.0008 hsa-miR-148b DMPK NM_001081563 3144 UCAGUGCAUC 3135 0.0008 hsa-miR-148b CD177 NM_020406 1832 UCAGUGCAUC 1823 0.0010 hsa-miR-148b HPS5 NM_181507 4873 UCAGUGCAUC 4864 0.0012 hsa-miR-148b C20orf118 NM_080628 1218 UCAGUGCAUC 1209 0.0013 hsa-miR-148b DNMT1 NM_001130823 5134 UCAGUGCAU 5126 0.0013 hsa-miR-148b KIAA1147 NM_001080392 5121 UCAGUGCAUCA 5111 0.0014 hsa-miR-148b TMEM187 NM_003492 1429 UCAGUGCAU 1421 0.0014 hsa-miR-148b IDH3G NM_174869 1346 UCAGUGCAU 1338 0.0015 hsa-miR-148b PNPLA2 NM_020376 1854 UCAGUGCAU 1846 0.0015 hsa-miR-148b NCLN NM_020170 3657 UCAGUGCAUC 3648 0.0018 hsa-miR-148b USP7 NM_003470 4138 UCAGUGCAUC 4129 0.0018 hsa-miR-148b ABCB7 NM_004299 2340 UCAGUGCA 2333 0.0018 hsa-miR-148b TRAF3IP1 NM_015650 3098 UCAGUGCAUC 3089 0.0019 hsa-miR-148b KIAA1787 NM_032442 4954 UCAGUGCAU 4946 0.0020 hsa-miR-148b STAP1 NM_012108 1219 UCAGUGCAU 1211 0.0021 hsa-miR-148b MRPL37 NM_016491 1482 UCAGUGCA 1475 0.0021 hsa-miR-148b ATP4B NM_000705 1319 UCAGUGCAU 1311 0.0022 hsa-miR-148b CCKBR NM_176875 1586 UCAGUGCAU 1578 0.0022 hsa-miR-148b ELAVL2 NM_004432 3138 UCAGUGCAUC 3129 0.0023 hsa-miR-148b AGBL2 NM_024783 2940 UCAGUGCAU 2932 0.0023 hsa-miR-148b TUBE1 NM_016262 1751 UCAGUGCAU 1743 0.0025 hsa-miR-148b SF3A2 NM_007165 1642 UCAGUGCA 1635 0.0025 hsa-miR-148b ST6GALNAC2 NM_006456 1394 UCAGUGCAU 1386 0.0028 hsa-miR-148b AHDC1 NM_001029882 6167 UCAGUGCAU 6159 0.0028 hsa-miR-148b EMG1 NM_006331 974 UCAGUGCA 967 0.0029 hsa-miR-148b LDB3 NM_007078 3415 UCAGUGCAUC 3406 0.0030 hsa-miR-148b PNPLA6 NM_006702 4421 UCAGUGCA 4414 0.0031 hsa-miR-148b SIRT4 NM_012240 1084 UCAGUGCA 1077 0.0032 hsa-miR-148b ATP6AP2 NM_005765 1439 UCAGUGCAU 1431 0.0034 hsa-miR-148b DOCK6 NM_020812 6345 UCAGUGCA 6338 0.0034 hsa-miR-148b ETS1 NM_005238 2078 UCAGUGCAUC 2069 0.0035 hsa-miR-148b PSMD1 NM_002807 3114 UCAGUGCA 3107 0.0036 hsa-miR-148b NLRC5 NM_032206 6413 UCAGUGCAU 6405 0.0038 hsa-miR-148b TSNARE1 NM_145003 1840 UCAGUGCA 1833 0.0039 hsa-miR-148b CRISP1 NM_001131 1736 UCAGUGCAU 1728 0.0040 hsa-miR-148b SOS1 NM_005633 4108 UCAGUGCAUC 4099 0.0041 hsa-miR-148b SNRPD1 NM_006938 1173 UCAGUGCAU 1165 0.0043 hsa-miR-148b CIITA NM_000246 4369 UCAGUGCAU 4361 0.0043 hsa-miR-148b TMEM54 NM_033504 949 UCAGUGCA 942 0.0043 hsa-miR-148b MMP10 NM_002425 1642 UCAGUGCA 1635 0.0044 hsa-miR-148b DEDD NM_032998 2147 UCAGUGCAU 2139 0.0045 hsa-miR-148b GOLT1A NM_198447 673 UCAGUGCA 666 0.0045 hsa-miR-148b YIF1A NM_020470 1088 UCAGUGC 1082 0.0045 hsa-miR-148b SFRS11 NM_004768 1975 UCAGUGCAU 1967 0.0046 hsa-miR-148b THSD3 NM_199265 2809 UCAGUGCAU 2801 0.0046 hsa-miR-148b EXOC4 NM_021807 4025 UCAGUGCAU 4017 0.0047 hsa-miR-148b MRPL28 NM_006428 883 UCAGUGCA 876 0.0047 hsa-miR-148b TAC4 NM_170685 566 UCAGUGCA 559 0.0051 hsa-miR-148b CD72 NM_001782 1222 UCAGUGCA 1215 0.0052 hsa-miR-148b SNAP91 NM_014841 3742 UCAGUGCAU 3734 0.0053 hsa-miR-148b ITGA11 NM_001004439 4743 UCAGUGCAU 4735 0.0053 hsa-miR-148b COL4A1 NM_001845 5405 UCAGUGCAU 5397 0.0054 hsa-miR-148b RAB34 NM_031934 1473 UCAGUGCA 1466 0.0054 hsa-miR-148b DYRK1B NM_004714 2301 UCAGUGCA 2294 0.0056 hsa-miR-148b MRGPRX3 NM_054031 1576 UCAGUGCA 1569 0.0056 hsa-miR-148b ERRFI1 NM_018948 3015 UCAGUGCAU 3007 0.0057 hsa-miR-148b ABCD3 NM_002858 2986 UCAGUGCAU 2978 0.0058 hsa-miR-148b MRPS10 NM_018141 1215 UCAGUGCAU 1207 0.0058 hsa-miR-148b LRRC29 NM_012163 1408 UCAGUGCA 1401 0.0059 hsa-miR-148b DUSP28 NM_001033575 1206 UCAGUGCA 1199 0.0060 hsa-miR-148b RGMA NM_020211 1986 UCAGUGCAU 1978 0.0061 hsa-miR-148b WDFY4 NM_020945 9936 UCAGUGCA 9929 0.0061 hsa-miR-148b PHEX NM_000444 2514 UCAGUGCA 2507 0.0062 hsa-miR-148b KLF5 NM_001730 2455 UCAGUGCAU 2447 0.0063 hsa-miR-148b QARS NM_005051 2347 UCAGUGC 2341 0.0063 hsa-miR-148b C12orf59 NM_153022 1547 UCAGUGCAU 1539 0.0064 hsa-miR-148b OSBPL11 NM_022776 4565 UCAGUGCAU 4557 0.0064 hsa-miR-148b ZNF274 NM_133502 2749 UCAGUGCA 2742 0.0064 hsa-miR-148b ADRBK2 NM_005160 5757 UCAGUGCAUC 5748 0.0065 hsa-miR-148b CNTN4 NM_175607 3517 UCAGUGCAU 3509 0.0066 hsa-miR-148b RBMS2 NM_002898 8023 UCAGUGCAUC 8014 0.0067 hsa-miR-148b TRPM5 NM_014555 3573 UCAGUGCA 3566 0.0067 hsa-miR-148b COL2A1 NM_001844 4734 UCAGUGCA 4727 0.0067 hsa-miR-148b RASAL1 NM_004658 2909 UCAGUGCA 2902 0.0067 hsa-miR-148b VSIG1 NM_182607 2881 UCAGUGCAU 2873 0.0069 hsa-miR-148b YTHDC2 NM_022828 4537 UCAGUGCAU 4529 0.0069 hsa-miR-148b CAND1 NM_018448 4179 UCAGUGCAU 4171 0.0069 hsa-miR-148b INVS NM_014425 3624 UCAGUGCA 3617 0.0071 hsa-miR-148b GUCY2C NM_004963 3319 UCAGUGCA 3312 0.0072 hsa-miR-148b LOC389073 NM_001099334 881 UCAGUGCA 874 0.0072 hsa-miR-148b USP32 NM_032582 6967 UCAGUGCAU 6959 0.0073 hsa-miR-148b LRRC41 NM_006369 2626 UCAGUGCA 2619 0.0073 hsa-miR-148b NDUFS4 NM_002495 567 UCAGUGC 561 0.0073 hsa-miR-148b SYNC1 NM_030786 1854 UCAGUGCAU 1846 0.0074 hsa-miR-148b TSLP NM_033035 1811 UCAGUGCAU 1803 0.0074 hsa-miR-148b MTMR14 NM_001077525 2440 UCAGUGCA 2433 0.0074 hsa-miR-148b SLC12A7 NM_006598 4202 UCAGUGCAU 4194 0.0076 hsa-miR-148b CDC14A NM_003672 2470 UCAGUGCAU 2462 0.0077 hsa-miR-148b C8A NM_000562 1946 UCAGUGCA 1939 0.0077 hsa-miR-148b HOMER1 NM_004272 3617 UCAGUGCAU 3609 0.0078 hsa-miR-148b USP6 NM_004505 7774 UCAGUGCAU 7766 0.0078 hsa-miR-148b YWHAB NM_003404 2728 UCAGUGCAU 2720 0.0079 hsa-miR-148b EMP1 NM_001423 1064 UCAGUGCAU 1056 0.0079 hsa-miR-148b PKD2 NM_000297 3880 UCAGUGCAU 3872 0.0080 hsa-miR-148b NR2C2AP NM_176880 1069 UCAGUGCA 1062 0.0080 hsa-miR-148b ABI3 NM_016428 1637 UCAGUGCA 1630 0.0081 hsa-miR-148b MAP3K4 NM_005922 5360 UCAGUGCA 5353 0.0081 hsa-miR-148b KRR1 NM_007043 2226 UCAGUGCAU 2218 0.0082 hsa-miR-148b A4GNT NM_016161 1557 UCAGUGCA 1550 0.0083 hsa-miR-148b RNUXA NM_032177 1295 UCAGUGCA 1288 0.0083 hsa-miR-148b TMEM170 NM_145254 837 UCAGUGCA 830 0.0083 hsa-miR-148b LEPROTL1 NM_015344 2638 UCAGUGCAU 2630 0.0084 hsa-miR-148b IRX4 NM_016358 2103 UCAGUGCA 2096 0.0084 hsa-miR-148b MAF1 NM_032272 1723 UCAGUGCA 1716 0.0084 hsa-miR-148b NPEPL1 NM_024663 1720 UCAGUGCA 1713 0.0084 hsa-miR-148b VPS37A NM_152415 1675 UCAGUGCA 1668 0.0084 hsa-miR-148b ZPLD1 NM_175056 2598 UCAGUGCAU 2590 0.0085 hsa-miR-148b GADD45A NM_001924 832 UCAGUGCA 825 0.0085 hsa-miR-148b KRT76 NM_015848 2499 UCAGUGCA 2492 0.0086 hsa-miR-148b CABP7 NM_182527 2850 UCAGUGCAU 2842 0.0087 hsa-miR-148b SPON2 NM_012445 1457 UCAGUGCA 1450 0.0088 hsa-miR-148b LYSMD2 NM_153374 1298 UCAGUGCA 1291 0.0088 hsa-miR-148b LZTFL1 NM_020347 1283 UCAGUGCAU 1275 0.0090 hsa-miR-148b NUP133 NM_018230 3965 UCAGUGCA 3958 0.0092 hsa-miR-148b GLRX5 NM_016417 692 UCAGUGCA 685 0.0092 hsa-miR-148b TBP NM_003194 1443 UCAGUGCA 1436 0.0093 hsa-miR-148b TNIP2 NM_024309 1832 UCAGUGCA 1825 0.0094 hsa-miR-148b IL15 NM_172174 1650 UCAGUGCA 1643 0.0095 hsa-miR-148b IGSF21 NM_032880 1821 UCAGUGC 1815 0.0095 hsa-miR-148b NAT14 NM_020378 994 UCAGUGCA 987 0.0096 hsa-miR-148b BAI3 NM_001704 5063 UCAGUGCA 5056 0.0096 hsa-miR-148b C1GALT1 NM_020156 1362 UCAGUGCA 1355 0.0097 hsa-miR-148b ABHD12 NM_001042472 1647 UCAGUGCA 1640 0.0098 hsa-miR-148b GPBAR1 NM_001077191 1979 UCAGUGC 1973 0.0098 hsa-miR-148b C2orf15 NM_144706 1351 UCAGUGCA 1344 0.0099 hsa-miR-148b BRPF1 NM_001003694 4153 UCAGUGCA 4146 0.0101 hsa-miR-148b C15orf27 NM_152335 2397 UCAGUGCA 2390 0.0101 hsa-miR-148b PPP2R2C NM_020416 3062 UCAGUGCAU 3054 0.0103 hsa-miR-148b CDK5R1 NM_003885 3847 UCAGUGCAU 3839 0.0103 hsa-miR-148b TAGLN2 NM_003564 1177 UCAGUGCA 1170 0.0104 hsa-miR-148b CPT1A NM_001876 3779 UCAGUGCAU 3771 0.0105 hsa-miR-148b ASPH NM_004318 3928 UCAGUGCAU 3920 0.0106 hsa-miR-148b USF1 NM_007122 1324 UCAGUGCA 1317 0.0107 hsa-miR-148b TMEM49 NM_030938 1647 UCAGUGCA 1640 0.0107 hsa-miR-148b SNN NM_003498 593 UCAGUGCAU 585 0.0108 hsa-miR-148b ZNF223 NM_013361 2323 UCAGUGCA 2316 0.0108 hsa-miR-148b ZDHHC3 NM_016598 3374 UCAGUGCAUC 3365 0.0109 hsa-miR-148b RARRES2 NM_002889 563 UCAGUGC 557 0.0110 hsa-miR-148b MAML3 NM_018717 4659 UCAGUGCAU 4651 0.0111 hsa-miR-148b CLCN6 NM_001286 5341 UCAGUGCAU 5333 0.0111 hsa-miR-148b MARCH3 NM_178450 1291 UCAGUGCAU 1283 0.0112 hsa-miR-148b DRG2 NM_001388 1432 UCAGUGCA 1425 0.0112 hsa-miR-148b UGT1A8 NM_019076 2189 UCAGUGCA 2182 0.0112 hsa-miR-148b EPHA5 NM_004439 3221 UCAGUGC 3215 0.0113 hsa-miR-148b DNAJB12 NM_001002762 2612 UCAGUGCAU 2604 0.0114 hsa-miR-148b UGT1A6 NM_001072 2271 UCAGUGCA 2264 0.0115 hsa-miR-148b NCKIPSD NM_016453 2346 UCAGUGCA 2339 0.0115 hsa-miR-148b ITSN2 NM_006277 5845 UCAGUGCA 5838 0.0115 hsa-miR-148b FLJ20674 NM_019086 4697 UCAGUGCAU 4689 0.0116 hsa-miR-148b MLL NM_005933 14523 UCAGUGCAU 14515 0.0116 hsa-miR-148b C6orf97 NM_025059 4095 UCAGUGCAU 4087 0.0116 hsa-miR-148b SAPS1 NM_014931 3073 UCAGUGCA 3066 0.0116 hsa-miR-148b DEFB118 NM_054112 468 UCAGUGC 462 0.0116 hsa-miR-148b CECR6 NM_031890 2680 UCAGUGCAU 2672 0.0118 hsa-miR-148b MGLL NM_007283 3425 UCAGUGCAU 3417 0.0119 hsa-miR-148b SLC5A7 NM_021815 2407 UCAGUGCAU 2399 0.0119 hsa-miR-148b FAM105A NM_019018 1344 UCAGUGCA 1337 0.0119 hsa-miR-148b DUOXA2 NM_207581 1343 UCAGUGC 1337 0.0120 hsa-miR-148b ARG2 NM_001172 1198 UCAGUGCA 1191 0.0121 hsa-miR-148b WDR20 NM_181291 2183 UCAGUGCA 2176 0.0122 hsa-miR-148b CHCHD3 NM_017812 1126 UCAGUGCA 1119 0.0122 hsa-miR-148b PPP1R7 NM_002712 1229 UCAGUGC 1223 0.0122 hsa-miR-148b PVALB NM_002854 393 UCAGUGC 387 0.0123 hsa-miR-148b TET2 NM_001127208 8636 UCAGUGCAU 8628 0.0124 hsa-miR-148b KCTD16 NM_020768 5096 UCAGUGCAU 5088 0.0124 hsa-miR-148b TGFB2 NM_003238 4205 UCAGUGCAU 4197 0.0124 hsa-miR-148b PTEN NM_000314 5401 UCAGUGCAU 5393 0.0126 hsa-miR-148b TRAK2 NM_015049 3251 UCAGUGCAU 3243 0.0126 hsa-miR-148b RAG1 NM_000448 5887 UCAGUGCAU 5879 0.0126 hsa-miR-148b GNS NM_002076 4153 UCAGUGCAU 4145 0.0126 hsa-miR-148b PIGS NM_033198 2380 UCAGUGCA 2373 0.0126 hsa-miR-148b B4GALT6 NM_004775 4710 UCAGUGCAU 4702 0.0128 hsa-miR-148b KRT85 NM_002283 1703 UCAGUGCA 1696 0.0128 hsa-miR-148b PRKAA1 NM_206907 3807 UCAGUGCAU 3799 0.0129 hsa-miR-148b CUL5 NM_003478 3410 UCAGUGCAU 3402 0.0129 hsa-miR-148b IL23R NM_144701 2756 UCAGUGCA 2749 0.0129 hsa-miR-148b C7orf49 NM_024033 760 UCAGUGCA 753 0.0129 hsa-miR-148b ARF4 NM_001660 1390 UCAGUGCA 1383 0.0131 hsa-miR-148b TMEM44 NM_138399 1571 UCAGUGCA 1564 0.0132 hsa-miR-148b ZNF544 NM_014480 3455 UCAGUGCA 3448 0.0135 hsa-miR-148b CSF1 NM_000757 2666 UCAGUGCA 2659 0.0136 hsa-miR-148b ZNF689 NM_138447 2642 UCAGUGCA 2635 0.0137 hsa-miR-148b KIAA0226 NM_014687 5793 UCAGUGCAU 5785 0.0138 hsa-miR-148b MOGAT3 NM_178176 1238 UCAGUGC 1232 0.0138 hsa-miR-148b PRRG1 NM_000950 2143 UCAGUGCAU 2135 0.0139 hsa-miR-148b FEZ2 NM_001042548 1535 UCAGUGCA 1528 0.0139 hsa-miR-148b CCT6A NM_001762 1948 UCAGUGCA 1941 0.0140 hsa-miR-148b TAX1BP1 NM_006024 2842 UCAGUGCA 2835 0.0142 hsa-miR-148b EDG8 NM_030760 1912 UCAGUGCA 1905 0.0142 hsa-miR-148b OTUD4 NM_001102653 4166 UCAGUGCAU 4158 0.0143 hsa-miR-148b LOC51035 NM_015853 1253 UCAGUGC 1247 0.0143 hsa-miR-148b ATP7A NM_000052 4706 UCAGUGCAU 4698 0.0145 hsa-miR-148b PBXIP1 NM_020524 2403 UCAGUGCA 2396 0.0145 hsa-miR-148b MATK NM_139355 1991 UCAGUGC 1985 0.0145 hsa-miR-148b HTT NM_002111 12350 UCAGUGCAU 12342 0.0148 hsa-miR-148b BBC3 NM_014417 1046 UCAGUGCA 1039 0.0148 hsa-miR-148b KCND3 NM_004980 2581 UCAGUGC 2575 0.0148 hsa-miR-148b ALX4 NM_021926 1549 UCAGUGC 1543 0.0149 hsa-miR-148b C18orf62 NM_001037331 1357 UCAGUGCA 1350 0.0150 hsa-miR-148b ENOPH1 NM_021204 1691 UCAGUGCA 1684 0.0150 hsa-miR-148b TEK NM_000459 4718 UCAGUGCA 4711 0.0151 hsa-miR-148b TTC32 NM_001008237 705 UCAGUGC 699 0.0151 hsa-miR-148b BIRC7 NM_139317 1189 UCAGUGC 1183 0.0153 hsa-miR-148b SLC15A4 NM_145648 1850 UCAGUGCA 1843 0.0154 hsa-miR-148b TMEM9B NM_020644 1279 UCAGUGCA 1272 0.0155 hsa-miR-148b NDP NM_000266 1652 UCAGUGCA 1645 0.0156 hsa-miR-148b ESCO1 NM_052911 4337 UCAGUGCA 4330 0.0158 hsa-miR-148b OTX2 NM_021728 1938 UCAGUGCA 1931 0.0158 hsa-miR-148b C1orf190 NM_001013615 919 UCAGUGCA 912 0.0159 hsa-miR-148b ITGA5 NM_002205 4127 UCAGUGCA 4120 0.0159 hsa-miR-148b WNT1 NM_005430 2006 UCAGUGCA 1999 0.0160 hsa-miR-148b XPNPEP2 NM_003399 2344 UCAGUGCA 2337 0.0161 hsa-miR-148b NPTN NM_012428 2132 UCAGUGCA 2125 0.0161 hsa-miR-148b ZDHHC24 NM_207340 1311 UCAGUGC 1305 0.0162 hsa-miR-148b MYLK4 NM_001012418 3843 UCAGUGCAU 3835 0.0163 hsa-miR-148b TMEM132B NM_052907 7356 UCAGUGCAU 7348 0.0164 hsa-miR-148b ZNF490 NM_020714 4008 UCAGUGCAU 4000 0.0166 hsa-miR-148b IKZF1 NM_006060 3929 UCAGUGCAU 3921 0.0169 hsa-miR-148b USP48 NM_032236 3528 UCAGUGCA 3521 0.0169 hsa-miR-148b ITFG3 NM_032039 1912 UCAGUGCA 1905 0.0169 hsa-miR-148b DPCR1 NM_080870 1178 UCAGUGCA 1171 0.0170 hsa-miR-148b AKTIP NM_001012398 1555 UCAGUGCA 1548 0.0171 hsa-miR-148b CLEC4D NM_080387 1130 UCAGUGCA 1123 0.0171 hsa-miR-148b C20orf112 NM_080616 6011 UCAGUGCAU 6003 0.0172 hsa-miR-148b C2orf50 NM_182500 1068 UCAGUGC 1062 0.0172 hsa-miR-148b USP4 NM_003363 3445 UCAGUGCA 3438 0.0174 hsa-miR-148b PPP1R10 NM_002714 3870 UCAGUGCA 3863 0.0176 hsa-miR-148b EIF2C1 NM_012199 7083 UCAGUGCAU 7075 0.0177 hsa-miR-148b HCG27 NM_181717 1072 UCAGUGCA 1065 0.0177 hsa-miR-148b GTF2H1 NM_005316 2809 UCAGUGCA 2802 0.0178 hsa-miR-148b MEOX2 NM_005924 1484 UCAGUGCA 1477 0.0179 hsa-miR-148b CDH4 NM_001794 2981 UCAGUGC 2975 0.0180 hsa-miR-148b MFSD9 NM_032718 2150 UCAGUGCA 2143 0.0183 hsa-miR-148b C20orf82 NM_080826 1463 UCAGUGCA 1456 0.0183 hsa-miR-148b MED12L NM_053002 6518 UCAGUGCA 6511 0.0184 hsa-miR-148b MLF1IP NM_024629 2264 UCAGUGCA 2257 0.0184 hsa-miR-148b SOS2 NM_006939 4404 UCAGUGCA 4397 0.0184 hsa-miR-148b C6orf136 NM_145029 1332 UCAGUGC 1326 0.0185 hsa-miR-148b MAF NM_001031804 3160 UCAGUGCAU 3152 0.0187 hsa-miR-148b CHUK NM_001278 2809 UCAGUGCA 2802 0.0188 hsa-miR-148b PFAS NM_012393 4668 UCAGUGCA 4661 0.0188 hsa-miR-148b LIPA NM_001127605 2068 UCAGUGCA 2061 0.0192 hsa-miR-148b MMP13 NM_002427 1952 UCAGUGCA 1945 0.0193 hsa-miR-148b DOCK9 NM_001130048 6861 UCAGUGCA 6854 0.0193 hsa-miR-148b NPTX2 NM_002523 2695 UCAGUGCA 2688 0.0194 hsa-miR-148b ABLIM1 NM_001003408 4457 UCAGUGCAU 4449 0.0195 hsa-miR-148b HAVCR2 NM_032782 1213 UCAGUGCA 1206 0.0195 hsa-miR-148b ERBB3 NM_001982 5306 UCAGUGCA 5299 0.0195 hsa-miR-148b CXorf23 NM_198279 2643 UCAGUGCA 2636 0.0197 hsa-miR-148b NR1I2 NM_003889 3386 UCAGUGCA 3379 0.0197 hsa-miR-148b RNF219 NM_024546 3445 UCAGUGCA 3438 0.0198 hsa-miR-148b SACS NM_014363 14929 UCAGUGCA 14922 0.0198 hsa-miR-148b SERPINA4 NM_006215 1510 UCAGUGC 1504 0.0198 hsa-miR-148b WDR85 NM_138778 1633 UCAGUGC 1627 0.0199 hsa-miR-148b SLC7A2 NM_003046 7130 UCAGUGCAU 7122 0.0205 hsa-miR-148b NR2E1 NM_003269 2093 UCAGUGCA 2086 0.0205 hsa-miR-148b SLC22A5 NM_003060 2536 UCAGUGCA 2529 0.0205 hsa-miR-148b TTLL12 NM_015140 2243 UCAGUGCA 2236 0.0209 hsa-miR-148b EDG1 NM_001400 1930 UCAGUGCA 1923 0.0209 hsa-miR-148b TREML4 NM_198153 976 UCAGUGCA 969 0.0209 hsa-miR-148b HOXC8 NM_022658 2083 UCAGUGCA 2076 0.0209 hsa-miR-148b GPR52 NM_005684 1380 UCAGUGC 1374 0.0210 hsa-miR-148b SIGLEC8 NM_014442 2531 UCAGUGCA 2524 0.0211 hsa-miR-148b JMY NM_152405 3893 UCAGUGCAU 3885 0.0212 hsa-miR-148b ZNF30 NM_001099438 2570 UCAGUGC 2564 0.0212 hsa-miR-148b DEGS2 NM_206918 1128 UCAGUGC 1122 0.0212 hsa-miR-148b SLC25A43 NM_145305 1397 UCAGUGCA 1390 0.0214 hsa-miR-148b ELF5 NM_198381 2187 UCAGUGCA 2180 0.0217 hsa-miR-148b SLC25A28 NM_031212 1235 UCAGUGC 1229 0.0217 hsa-miR-148b PPAP2C NM_177543 1118 UCAGUGC 1112 0.0218 hsa-miR-148b MMP15 NM_002428 4417 UCAGUGCA 4410 0.0222 hsa-miR-148b PLEKHA7 NM_175058 4127 UCAGUGCA 4120 0.0222 hsa-miR-148b BCCIP NM_078469 1013 UCAGUGCA 1006 0.0222 hsa-miR-148b SETD4 NM_017438 2340 UCAGUGCA 2333 0.0222 hsa-miR-148b IKBKB NM_001556 2602 UCAGUGCA 2595 0.0222 hsa-miR-148b EDC4 NM_014329 4411 UCAGUGC 4405 0.0222 hsa-miR-148b LTK NM_002344 2811 UCAGUGC 2805 0.0223 hsa-miR-148b CKAP2 NM_001098525 2849 UCAGUGCA 2842 0.0224 hsa-miR-148b SIM1 NM_005068 3203 UCAGUGCA 3196 0.0224 hsa-miR-148b FLJ10769 NM_018210 1665 UCAGUGCA 1658 0.0225 hsa-miR-148b PIK3IP1 NM_052880 1647 UCAGUGCA 1640 0.0226 hsa-miR-148b RHD NM_016124 2532 UCAGUGCA 2525 0.0229 hsa-miR-148b MAP2K5 NM_145160 2019 UCAGUGC 2013 0.0230 hsa-miR-148b CTDSP1 NM_021198 1301 UCAGUGCA 1294 0.0231 hsa-miR-148b SLC25A39 NM_016016 1262 UCAGUGC 1256 0.0231 hsa-miR-148b C3orf21 NM_152531 2349 UCAGUGCA 2342 0.0232 hsa-miR-148b PQLC1 NM_025078 2134 UCAGUGCA 2127 0.0232 hsa-miR-148b TMPRSS12 NM_182559 1267 UCAGUGC 1261 0.0232 hsa-miR-148b ATP2A3 NM_174953 3730 UCAGUGCA 3723 0.0233 hsa-miR-148b SLC26A3 NM_000111 2766 UCAGUGC 2760 0.0234 hsa-miR-148b FAM55C NM_001134456 3124 UCAGUGCAU 3116 0.0237 hsa-miR-148b PRR18 NM_175922 2513 UCAGUGCA 2506 0.0237 hsa-miR-148b PSMD9 NM_002813 1264 UCAGUGCA 1257 0.0237 hsa-miR-148b FBXL19 NM_001099784 2420 UCAGUGCA 2413 0.0237 hsa-miR-148b RASA3 NM_007368 4091 UCAGUGCA 4084 0.0239 hsa-miR-148b ANKS3 NM_133450 2464 UCAGUGC 2458 0.0239 hsa-miR-148b C1orf212 NM_138428 1150 UCAGUGCA 1143 0.0240 hsa-miR-148b ARL6IP1 NM_015161 1970 UCAGUGCA 1963 0.0241 hsa-miR-148b RTKN NM_033046 2532 UCAGUGC 2526 0.0241 hsa-miR-148b FLJ13305 NM_032180 2949 UCAGUGCA 2942 0.0242 hsa-miR-148b PPP1R9B NM_032595 3082 UCAGUGCA 3075 0.0243 hsa-miR-148b GPR132 NM_013345 2111 UCAGUGCA 2104 0.0243 hsa-miR-148b PRNP NM_000311 2573 UCAGUGCA 2566 0.0244 hsa-miR-148b ATP6V1G3 NM_133326 657 UCAGUGC 651 0.0244 hsa-miR-148b ATAD3C NM_001039211 2932 UCAGUGCA 2925 0.0245 hsa-miR-148b MAFF NM_012323 1186 UCAGUGCA 1179 0.0246 hsa-miR-148b FAM73B NM_032809 3559 UCAGUGCA 3552 0.0246 hsa-miR-148b ZNF695 NM_020394 2619 UCAGUGCA 2612 0.0250 hsa-miR-148b IGF1 NM_001111283 6648 UCAGUGCAU 6640 0.0251 hsa-miR-148b SLC2A1 NM_006516 2653 UCAGUGCA 2646 0.0254 hsa-miR-148b ARFIP1 NM_001025595 1698 UCAGUGCA 1691 0.0255 hsa-miR-148b MPL NM_005373 2104 UCAGUGCA 2097 0.0255 hsa-miR-148b IKIP NM_153687 2869 UCAGUGCA 2862 0.0255 hsa-miR-148b LAMA3 NM_198129 10134 UCAGUGC 10128 0.0255 hsa-miR-148b CCRL2 NM_003965 1462 UCAGUGC 1456 0.0256 hsa-miR-148b ZNF445 NM_181489 10119 UCAGUGCAU 10111 0.0257 hsa-miR-148b SESN2 NM_031459 1979 UCAGUGCA 1972 0.0257 hsa-miR-148b MLLT10 NM_004641 3665 UCAGUGCA 3658 0.0257 hsa-miR-148b MMD NM_012329 1108 UCAGUGCA 1101 0.0259 hsa-miR-148b KIAA1274 NM_014431 3799 UCAGUGCA 3792 0.0263 hsa-miR-148b ACSL3 NM_004457 3635 UCAGUGCA 3628 0.0264 hsa-miR-148b SCAMP4 NM_079834 2393 UCAGUGCA 2386 0.0264 hsa-miR-148b NOL4 NM_003787 3345 UCAGUGCA 3338 0.0264 hsa-miR-148b PAPSS2 NM_001015880 3322 UCAGUGCA 3315 0.0264 hsa-miR-148b PADI1 NM_013358 3594 UCAGUGCA 3587 0.0265 hsa-miR-148b SBF2 NM_030962 6506 UCAGUGCA 6499 0.0265 hsa-miR-148b RABL5 NM_001130821 1897 UCAGUGCA 1890 0.0266 hsa-miR-148b JARID2 NM_004973 5427 UCAGUGCA 5420 0.0267 hsa-miR-148b RNASEL NM_021133 3074 UCAGUGCA 3067 0.0267 hsa-miR-148b GAS2L1 NM_152237 2506 UCAGUGCA 2499 0.0268 hsa-miR-148b TANC1 NM_033394 6938 UCAGUGCA 6931 0.0269 hsa-miR-148b LBR NM_002296 2086 UCAGUGCA 2079 0.0273 hsa-miR-148b FOXRED1 NM_017547 1810 UCAGUGC 1804 0.0276 hsa-miR-148b DHX15 NM_001358 2980 UCAGUGC 2974 0.0279 hsa-miR-148b SEMA4F NM_004263 3629 UCAGUGCA 3622 0.0281 hsa-miR-148b PPM1L NM_139245 2935 UCAGUGCA 2928 0.0282 hsa-miR-148b ANKRD13A NM_033121 3360 UCAGUGCA 3353 0.0284 hsa-miR-148b ZDHHC7 NM_017740 1794 UCAGUGCA 1787 0.0285 hsa-miR-148b TMEM97 NM_014573 2358 UCAGUGCA 2351 0.0287 hsa-miR-148b SPTLC3 NM_018327 3345 UCAGUGCA 3338 0.0287 hsa-miR-148b STX3 NM_004177 1087 UCAGUGCA 1080 0.0288 hsa-miR-148b ZFYVE26 NM_015346 9411 UCAGUGCA 9404 0.0288 hsa-miR-148b UBE3B NM_130466 4813 UCAGUGCA 4806 0.0292 hsa-miR-148b PHF21B NM_138415 3118 UCAGUGCA 3111 0.0293 hsa-miR-148b INHBB NM_002193 2736 UCAGUGCA 2729 0.0293 hsa-miR-148b TRPM1 NM_002420 5260 UCAGUGC 5254 0.0293 hsa-miR-148b STXBP1 NM_003165 3499 UCAGUGCA 3492 0.0296 hsa-miR-148b SERINC2 NM_178865 1886 UCAGUGC 1880 0.0296 hsa-miR-148b UFD1L NM_005659 1117 UCAGUGC 1111 0.0296 hsa-miR-148b GPM6A NM_201591 3034 UCAGUGCA 3027 0.0297 hsa-miR-148b EIF2C4 NM_017629 2910 UCAGUGCA 2903 0.0298 hsa-miR-148b ETV7 NM_016135 1592 UCAGUGC 1586 0.0298 hsa-miR-148b PKNOX2 NM_022062 2093 UCAGUGCA 2086 0.0300 hsa-miR-148b SF3B3 NM_012426 4044 UCAGUGC 4038 0.0302 hsa-miR-148b NUDT12 NM_031438 2266 UCAGUGCA 2259 0.0304 hsa-miR-148b HECW2 NM_020760 5876 UCAGUGCA 5869 0.0304 hsa-miR-148b ZCCHC2 NM_017742 4793 UCAGUGCA 4786 0.0304 hsa-miR-148b MAFB NM_005461 2733 UCAGUGCA 2726 0.0304 hsa-miR-148b PNOC NM_006228 872 UCAGUGC 866 0.0304 hsa-miR-148b KLK15 NM_017509 1309 UCAGUGC 1303 0.0305 hsa-miR-148b CECR5 NM_033070 1672 UCAGUGC 1666 0.0306 hsa-miR-148b HSPA4L NM_014278 3209 UCAGUGC 3203 0.0307 hsa-miR-148b MYO19 NM_025109 2943 UCAGUGC 2937 0.0308 hsa-miR-148b NID1 NM_002508 4754 UCAGUGCA 4747 0.0309 hsa-miR-148b CAST NM_001750 3673 UCAGUGCA 3666 0.0310 hsa-miR-148b PIGA NM_002641 2935 UCAGUGCA 2928 0.0310 hsa-miR-148b NOTUM NM_178493 1794 UCAGUGC 1788 0.0311 hsa-miR-148b EPAS1 NM_001430 4254 UCAGUGCA 4247 0.0313 hsa-miR-148b ADPRH NM_001125 2191 UCAGUGCA 2184 0.0314 hsa-miR-148b EPM2A NM_005670 1713 UCAGUGCA 1706 0.0318 hsa-miR-148b TOMM70A NM_014820 4286 UCAGUGCA 4279 0.0318 hsa-miR-148b SMOC1 NM_001034852 2559 UCAGUGCA 2552 0.0319 hsa-miR-148b VPS45 NM_007259 2774 UCAGUGC 2768 0.0319 hsa-miR-148b GOLPH3L NM_018178 2562 UCAGUGCA 2555 0.0320 hsa-miR-148b C5orf30 NM_033211 1415 UCAGUGCA 1408 0.0321 hsa-miR-148b S100Z NM_130772 729 UCAGUGC 723 0.0321 hsa-miR-148b EXOSC1 NM_016046 1017 UCAGUGC 1011 0.0322 hsa-miR-148b PDK4 NM_002612 2323 UCAGUGCA 2316 0.0323 hsa-miR-148b ATG9B NM_173681 2846 UCAGUGCA 2839 0.0323 hsa-miR-148b CNN1 NM_001299 1314 UCAGUGC 1308 0.0325 hsa-miR-148b SFMBT1 NM_001005159 3607 UCAGUGC 3601 0.0325 hsa-miR-148b PAG1 NM_018440 4514 UCAGUGCAU 4506 0.0328 hsa-miR-148b RPGRIP1L NM_015272 6024 UCAGUGCA 6017 0.0328 hsa-miR-148b FAM104A NM_001098832 1189 UCAGUGCA 1182 0.0329 hsa-miR-148b SNF1LK NM_173354 2645 UCAGUGCA 2638 0.0334 hsa-miR-148b JUB NM_032876 3015 UCAGUGCA 3008 0.0336 hsa-miR-148b MAP3K9 NM_033141 3473 UCAGUGCA 3466 0.0337 hsa-miR-148b GALNTL2 NM_054110 3592 UCAGUGCA 3585 0.0337 hsa-miR-148b DSC2 NM_004949 3802 UCAGUGCA 3795 0.0339 hsa-miR-148b MREG NM_018000 2406 UCAGUGCA 2399 0.0341 hsa-miR-148b FMR1 NM_002024 2851 UCAGUGCA 2844 0.0342 hsa-miR-148b GAPVD1 NM_015635 5058 UCAGUGCA 5051 0.0342 hsa-miR-148b HTRA2 NM_013247 2108 UCAGUGC 2102 0.0343 hsa-miR-148b C5orf25 NM_198567 1534 UCAGUGC 1528 0.0343 hsa-miR-148b GPATCH8 NM_001002909 5383 UCAGUGCA 5376 0.0345 hsa-miR-148b STT3A NM_152713 2262 UCAGUGC 2256 0.0345 hsa-miR-148b CDC2L5 NM_003718 5237 UCAGUGC 5231 0.0345 hsa-miR-148b MYO3A NM_017433 5309 UCAGUGC 5303 0.0352 hsa-miR-148b AMOTL2 NM_016201 4489 UCAGUGCA 4482 0.0353 hsa-miR-148b SLC26A4 NM_000441 2980 UCAGUGCA 2973 0.0354 hsa-miR-148b ST18 NM_014682 6113 UCAGUGCA 6106 0.0355 hsa-miR-148b VPS24 NM_016079 1839 UCAGUGCA 1832 0.0356 hsa-miR-148b TUBA8 NM_018943 1937 UCAGUGC 1931 0.0357 hsa-miR-148b CALR NM_004343 1757 UCAGUGC 1751 0.0357 hsa-miR-148b KCNAB1 NM_172159 4128 UCAGUGCA 4121 0.0358 hsa-miR-148b LRPPRC NM_133259 6019 UCAGUGCA 6012 0.0359 hsa-miR-148b ZNF498 NM_145115 3752 UCAGUGCA 3745 0.0359 hsa-miR-148b NT5C3 NM_001002009 1713 UCAGUGC 1707 0.0360 hsa-miR-148b ZDHHC22 NM_174976 2939 UCAGUGCA 2932 0.0361 hsa-miR-148b KIF2C NM_006845 2723 UCAGUGC 2717 0.0361 hsa-miR-148b HMOX1 NM_002133 1049 UCAGUGC 1043 0.0361 hsa-miR-148b TNRC6A NM_014494 6403 UCAGUGCA 6396 0.0365 hsa-miR-148b ZNF354C NM_014594 1943 UCAGUGC 1937 0.0365 hsa-miR-148b H2AFY NM_138610 1519 UCAGUGC 1513 0.0366 hsa-miR-148b NSMAF NM_003580 3285 UCAGUGC 3279 0.0368 hsa-miR-148b STARD13 NM_178006 4486 UCAGUGCA 4479 0.0369 hsa-miR-148b EPN2 NM_014964 4648 UCAGUGCA 4641 0.0369 hsa-miR-148b CD68 NM_001251 1284 UCAGUGC 1278 0.0369 hsa-miR-148b ALCAM NM_001627 2571 UCAGUGCA 2564 0.0370 hsa-miR-148b CFL2 NM_138638 2494 UCAGUGCA 2487 0.0371 hsa-miR-148b TRIM59 NM_173084 3349 UCAGUGCA 3342 0.0371 hsa-miR-148b RASGRP1 NM_005739 4139 UCAGUGCA 4132 0.0371 hsa-miR-148b VGLL4 NM_001128219 3489 UCAGUGCA 3482 0.0372 hsa-miR-148b LDLR NM_000527 4729 UCAGUGCA 4722 0.0376 hsa-miR-148b GPR180 NM_180989 2708 UCAGUGCA 2701 0.0377 hsa-miR-148b TGIF2 NM_021809 3182 UCAGUGCA 3175 0.0377 hsa-miR-148b ATP10A NM_024490 4927 UCAGUGC 4921 0.0377 hsa-miR-148b C3orf58 NM_173552 1953 UCAGUGCA 1946 0.0380 hsa-miR-148b ZBTB7A NM_015898 3985 UCAGUGCA 3978 0.0381 hsa-miR-148b CAT NM_001752 2230 UCAGUGC 2224 0.0382 hsa-miR-148b AMZ1 NM_133463 2745 UCAGUGCA 2738 0.0383 hsa-miR-148b PITPNM2 NM_020845 4661 UCAGUGCA 4654 0.0384 hsa-miR-148b SLC25A18 NM_031481 1875 UCAGUGC 1869 0.0386 hsa-miR-148b NRP1 NM_003873 5437 UCAGUGCA 5430 0.0389 hsa-miR-148b OXSR1 NM_005109 2920 UCAGUGCA 2913 0.0389 hsa-miR-148b DGCR2 NM_005137 2439 UCAGUGCA 2432 0.0389 hsa-miR-148b SNRK NM_017719 3171 UCAGUGCA 3164 0.0390 hsa-miR-148b GPR137C NM_001099652 3523 UCAGUGCA 3516 0.0392 hsa-miR-148b SNX21 NM_001042633 2114 UCAGUGCA 2107 0.0392 hsa-miR-148b FLJ31818 NM_152556 2664 UCAGUGCA 2657 0.0392 hsa-miR-148b EMID1 NM_133455 1718 UCAGUGC 1712 0.0392 hsa-miR-148b OS9 NM_006812 2446 UCAGUGC 2440 0.0394 hsa-miR-148b WNT10A NM_025216 2276 UCAGUGC 2270 0.0394 hsa-miR-148b DUSP1 NM_004417 1857 UCAGUGC 1851 0.0397 hsa-miR-148b ADAMTSL2 NM_014694 3194 UCAGUGC 3188 0.0400 hsa-miR-148b PLB1 NM_153021 4884 UCAGUGC 4878 0.0410 hsa-miR-148b CACNA1E NM_000721 8769 UCAGUGCA 8762 0.0412 hsa-miR-148b COL9A2 NM_001852 2320 UCAGUGC 2314 0.0413 hsa-miR-148b MTMR12 NM_001040446 4226 UCAGUGCA 4219 0.0414 hsa-miR-148b ZFAT NM_001029939 4516 UCAGUGCA 4509 0.0417 hsa-miR-148b RSU1 NM_012425 3387 UCAGUGCA 3380 0.0417 hsa-miR-148b UBE2L6 NM_198183 949 UCAGUGC 943 0.0419 hsa-miR-148b CD40 NM_001250 1014 UCAGUGC 1008 0.0421 hsa-miR-148b CIAO1 NM_004804 2770 UCAGUGCA 2763 0.0422 hsa-miR-148b DYRK1A NM_130436 4496 UCAGUGCA 4489 0.0425 hsa-miR-148b AK2 NM_013411 2560 UCAGUGCA 2553 0.0425 hsa-miR-148b C1D NM_006333 674 UCAGUGC 668 0.0425 hsa-miR-148b KCTD2 NM_015353 1618 UCAGUGCA 1611 0.0426 hsa-miR-148b MNT NM_020310 4822 UCAGUGCA 4815 0.0428 hsa-miR-148b HEPH NM_001130860 4333 UCAGUGC 4327 0.0428 hsa-miR-148b PDZD11 NM_016484 1221 UCAGUGC 1215 0.0431 hsa-miR-148b ZADH2 NM_175907 2801 UCAGUGCA 2794 0.0432 hsa-miR-148b GTPBP1 NM_004286 3458 UCAGUGCA 3451 0.0434 hsa-miR-148b ZBED4 NM_014838 4833 UCAGUGCA 4826 0.0434 hsa-miR-148b MFGE8 NM_005928 1521 UCAGUGC 1515 0.0435 hsa-miR-148b BTBD3 NM_014962 4616 UCAGUGCA 4609 0.0437 hsa-miR-148b LMBRD2 NM_001007527 2631 UCAGUGC 2625 0.0438 hsa-miR-148b ATP2B3 NM_021949 4916 UCAGUGCA 4909 0.0439 hsa-miR-148b ACO1 NM_002197 2954 UCAGUGC 2948 0.0439 hsa-miR-148b SLC7A5 NM_003486 2049 UCAGUGCA 2042 0.0440 hsa-miR-148b PPP2R2A NM_002717 2315 UCAGUGC 2309 0.0441 hsa-miR-148b GLCE NM_015554 3617 UCAGUGCA 3610 0.0442 hsa-miR-148b TMEM165 NM_018475 1353 UCAGUGC 1347 0.0442 hsa-miR-148b STEAP4 NM_024636 3436 UCAGUGCA 3429 0.0443 hsa-miR-148b DDAH1 NM_001134445 3944 UCAGUGCA 3937 0.0443 hsa-miR-148b GLIPR1 NM_006851 3506 UCAGUGCA 3499 0.0444 hsa-miR-148b C18orf10 NM_015476 1630 UCAGUGC 1624 0.0446 hsa-miR-148b RPS6KA4 NM_003942 2729 UCAGUGC 2723 0.0446 hsa-miR-148b CANX NM_001746 3507 UCAGUGCA 3500 0.0447 hsa-miR-148b CYP26B1 NM_019885 2216 UCAGUGCA 2209 0.0447 hsa-miR-148b ERC2 NM_015576 5043 UCAGUGCA 5036 0.0449 hsa-miR-148b UGT8 NM_001128174 2374 UCAGUGC 2368 0.0452 hsa-miR-148b SYNJ1 NM_203446 4913 UCAGUGCA 4906 0.0454 hsa-miR-148b C6orf47 NM_021184 2345 UCAGUGC 2339 0.0456 hsa-miR-148b ATXN7L1 NM_020725 4195 UCAGUGCA 4188 0.0459 hsa-miR-148b C14orf79 NM_174891 1536 UCAGUGC 1530 0.0463 hsa-miR-148b ROBO4 NM_019055 3612 UCAGUGC 3606 0.0464 hsa-miR-148b CABLES1 NM_001100619 3462 UCAGUGCA 3455 0.0465 hsa-miR-148b SPIRE1 NM_001128626 2532 UCAGUGCA 2525 0.0466 hsa-miR-148b ADCY9 NM_001116 5264 UCAGUGCA 5257 0.0467 hsa-miR-148b ZNF434 NM_017810 2545 UCAGUGC 2539 0.0467 hsa-miR-148b KIAA1430 NM_020827 2351 UCAGUGCA 2344 0.0469 hsa-miR-148b CDH22 NM_021248 3468 UCAGUGC 3462 0.0469 hsa-miR-148b MMP11 NM_005940 1881 UCAGUGC 1875 0.0469 hsa-miR-148b PTGS1 NM_000962 2508 UCAGUGCA 2501 0.0470 hsa-miR-148b RAB40B NM_006822 894 UCAGUGC 888 0.0471 hsa-miR-148b LASS5 NM_147190 1310 UCAGUGC 1304 0.0471 hsa-miR-148b FAM100B NM_182565 1235 UCAGUGC 1229 0.0472 hsa-miR-148b PIK3C2A NM_002645 7847 UCAGUGCA 7840 0.0473 hsa-miR-148b PSCD3 NM_004227 1953 UCAGUGCA 1946 0.0473 hsa-miR-148b LYSMD3 NM_198273 3975 UCAGUGCA 3968 0.0475 hsa-miR-148b CENPF NM_016343 9707 UCAGUGC 9701 0.0475 hsa-miR-148b PLA2G4F NM_213600 2995 UCAGUGC 2989 0.0477 hsa-miR-148b ITGA9 NM_002207 3569 UCAGUGC 3563 0.0477 hsa-miR-148b KIAA0232 NM_014743 6458 UCAGUGCA 6451 0.0480 hsa-miR-148b ORMDL2 NM_014182 599 UCAGUGC 593 0.0481 hsa-miR-148b AP4E1 NM_007347 6518 UCAGUGCA 6511 0.0482 hsa-miR-148b UBA6 NM_018227 3362 UCAGUGCA 3355 0.0484 hsa-miR-148b HCFC2 NM_013320 5069 UCAGUGCA 5062 0.0485 hsa-miR-148b MRAS NM_012219 1262 UCAGUGCA 1255 0.0486 hsa-miR-148b SGCB NM_000232 1937 UCAGUGCA 1930 0.0488 hsa-miR-148b C2orf56 NM_144736 1992 UCAGUGC 1986 0.0488 hsa-miR-148b ZNRF1 NM_032268 1469 UCAGUGCA 1462 0.0490 hsa-miR-148b GRPEL1 NM_025196 1256 UCAGUGC 1250 0.0490 hsa-miR-148b DMRTB1 NM_033067 1809 UCAGUGC 1803 0.0490 hsa-miR-148b ARSD NM_001669 3458 UCAGUGCA 3451 0.0491 hsa-miR-148b GRM6 NM_000843 4854 UCAGUGCA 4847 0.0495 hsa-miR-148b SMTNL2 NM_001114974 1656 UCAGUGC 1650 0.0495 hsa-miR-148b LSM8 NM_016200 981 UCAGUGC 975 0.0496 hsa-miR-148b EPHA4 NM_004438 4077 UCAGUGCA 4070 0.0500 hsa-miR-148b CCDC40 NM_017950 4076 UCAGUGC 4070 0.0500 Supplementary Table 2. Predicted target of miR148b with pvalue ≤ 0.05.