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Altered DNA methylation associated with nervosa anorexia in males

Artem Kim

Institute of Genetics and Development of Rennes (Igdr), Rennes, France

E-mail : bhuvaneswari.bibleraaj@uhsm.nhs.uk

Brigitte Izac

Cochin Institute, Genomics Platform, Inserm U1016, Paris, France

Nicolas Lebrun

Institute of Psychiatry and Neuroscience of Paris (IPNP), Inserm U1266, Paris, France

Nicolas Ramoz

Institute of Psychiatry and Neuroscience of Paris (IPNP), Inserm U1266, Paris, France

Corinne Blanchet

Teenagers' House, Cochin Hospital, Paris, France

Franck Letourneur

Cochin Institute, Genomics Platform, Inserm U1016, Paris, France

Marie Rose Moro

Teenagers' House, Cochin Hospital, Paris, France

Philip Gorwood

Institute of Psychiatry and Neuroscience of Paris (IPNP), Inserm U1266, Paris, France

CMME, St. Anne's Hospital, University Paris-Descartes, France.

Marie De Tayrac

Institute of Genetics and Development of Rennes (Igdr), Rennes, France

Thierry Bienvenu

Institute of Psychiatry and Neuroscience of Paris (IPNP), Inserm U1266, Paris, France

Laboratory of Molecular Genetics and Biology, Hôpital Cochin, HUPC, AP-HP, Paris, France

DOI: 10.15761/IMM.1000376

Article
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Abstract

Purpose: Anorexia nervosa (AN) is a serious psychiatric disorder characterized by abnormal eating behaviors, resulting in weight loss and increased mortality. Although more common in females, an estimated 5 to 10% of affected patients are males. Up to now, the exact cause of male AN is unknown. As with many psychiatric diseases, it's probably a combination of genetic, biological, psychological and environmental factors. Here, we used whole-genome bisulfite sequencing to determine the methylome of male individuals with AN.

Methods: We analyzed by bisulfite sequencing 3,340,894 biologically relevant CpG sites (Illumina TruSeqMethyl Capture EPIC kit) of 6 male patients affected with AN restrictive type. To reduce the environment effect, 4 related unaffected individuals were selected as controls.

Results: Comparisons between male patients affected with AN restrictive type and unaffected controls showed 153 differentially methylated regions and 1812 differentially methylated CpGs that corresponded to genes relevant to metabolic and nutritional status, psychiatric status and immune function. Moreover, the String network analysis software identified a subnetwork, related to MAPK signaling pathway, PI3K-Akt signaling pathway and neurotrophin signaling pathway.

Conclusions: Our findings replicate several results concerning several target genes such as PRKAG2, RPTOR, and ICAM5 previously identified in female AN, and identified novel signaling pathways involving PI3K-Akt and neurotrophin signaling pathway disturbed in AN.

Key words

anorexia nervosa, restrictive type, anorexia, whole-genome bisulfite sequencing

Introduction

Anorexia nervosa (AN) is a complex neuropsychiatric disorder characterized by weight loss, and an intense fear of gaining weight. Family and twin studies of AN have shown that genetic and environmental factors play important roles in the pathogenesis of AN. Twin studies have estimated the heritability to be~56% [1]. However, while several genes were identified by candidate gene studies and genome-wide association studies (GWAS), they often failed to replicate in other studies [2-5]. The same observation was found for major depression sharing common genetic and environmental risk factors with AN [6]. Indeed, young people who suffered from AN are at high risk of depression [7]. Epigenetics and environmental factors might play a crucial role in the development of AN as well as in major depressive disorder [8]. Recently, a genome-wide DNA methylation study was conducted in women with active AN, AN in remission and in non-AN controls. Global analysis of the 3 groups revealed 295 differentially methylated sites (DMS) representing 277 genes. Some of the identified genes were related to nutrition, to bone and tissue heath, immune function/inflammation, as well as general or transcriptional processes, and glia-neuron interaction [9].

As AN is nine times more often in females than in males, focusing on gender specificities might help to shed light on its overall nature. We therefore propose an analysis of several male AN patients, supporting in part the recent findings and identifying a novel disease-relevant pathway.

Patients and methods

We analyzed more than 3.3 M biologically relevant CpGs (Illumina TruSeqMethyl Capture EPIC kit) of 6 male patients affected with AN restrictive type. This approach captures 3,340,894 CpG sites including 26,981 CpG islands (107 Mb), including American College of Medical Genetics (ACMG) genes, coding genes known to be involved in cancer, coding exons from Ensemble 70, and 100 promoters defined as being of high interest and difficult to sequence. To reduce the environment effect, 4 related unaffected individuals- parents of two AN individuals - were selected as controls. Briefly, libraries were prepared according to manufacturer protocol (using the TrueSeq DNA Methylation Kit (cat. EGMK81312, Illumina Inc., San Diego, CA), the NEXTflex Bisulfite Sequencing Kit (cat. 5119–02) and the 24 NEXTflex Bisulfite Sequencing Barcodes (cat. 511913)). After PCR-amplification, the bisulfite-treated Libraries were clustered on a V3 paired-end read flow cell and sequenced for 100 cycles on an Illumina NextSeq500 System (Illumina). Fastq files were generated using Illumina’s software CASAVA v1.8.2. Raw fastq reads were processed by a custom pipeline that consists of: (i) filtering raw fastq reads for pass filter reads, (ii) trimming adapter sequence by Trim Galore, (iii) genomic alignments performed using Bismark to reference human genome hg19, and (iv) methylation calling by a Bismark script (https://github.com/nf-core/methylseq). The R package methyKit was used for analysis and annotation of DNA [10]. After adjustment for potential confounders and exclusion of age-related CpG, differences were shown between AN individuals and controls. We initially investigated the differentially methylated regions (DMRs) in AN using a computational algorithm MethylKit with a stringent statistical cutoff of q-value (FDR adjusted p-value<0.01) and a minimum 25% change in methylation between AN patients and controls.

Results

This approach allowed us to identify a total of 153 DMRs in AN (Table 1). Of these, 53 DMRs were hypomethylated and 100 were hypermethylated, indicating that more genic regions tended to be methylated in AN individuals, similar to previous observations [9]. We further examined the genomic distribution of differentially methylated CpGs (DMCs). A total of 1812 DMC sites were identified in patients with AN compared to controls (Supplementary Table 1). Among the 1812 DMCs, 748 were hypomethylated, while 1065 were hypermethylated in the AN samples. Among these 1812 DMCs, 216functional genic regions included at least 2 DMCs. Interestingly, when we compared our DMCs list with the short list of Steiger and colleagues, 32 genes were found in both lists (Supplementary Table 2) [9]. Moreover, among the 100 top-ranked differentially methylated positions identified in patients with major depressive disorder, six were also found in our study (GNG4, SLC39A12, CRTAC1, MYO7A, NTM and RMST), four of them playing a role in neurite extension and neurogenesis (GNG4, SLC39A12, NTM and RMST) [8].

Table 1. Identification of differentially methylated regions (DMR) was performed using the predefined regions of Illumina TruSeq Methylcapture EPIC kit presenting at least 25 % methylation difference between ANcases and controls (column Change) with a q-value (FDR corrected p-value) threshold of 0.01

DMR

Chr

Nb Genes

Genes

p value

q value

Change

chrX_114502896_114503051

chrX

0

-

2.22451E-15

3.71584E-14

59.78484933

chr14_54375093_54375328

chr14

1

AL138479.

2.65207E-24

1.18277E-22

48.00682879

chr8_80036078_80036321

chr8

0

-

2.44128E-36

2.55548E-34

46.08190008

chr11_79289117_79289197

chr11

0

-

2.09678E-48

3.89352E-46

44.01889299

chr2_165342265_165342368

chr2

1

GRB14

5.17941E-11

4.44995E-10

43.62139918

chr5_118811413_118811668

chr5

2

HSD17B4|snoU1

4.63731E-11

4.01928E-10

43.59922516

chr19_45113900_45114110

chr19

2

CEACAM22P|IGSF23

9.32156E-32

7.39337E-30

40.63822929

chr12_39885082_39885307

chr12

0

-

1.48466E-54

3.57503E-52

40.49521423

chr11_99564881_99564954

chr11

1

CNTN5

9.37129E-15

1.42916E-13

39.62334329

chr20_46228572_46228783

chr20

1

NCOA3

1.11712E-13

1.47266E-12

39.56217898

chr12_9507242_9507307

chr12

0

-

1.05726E-24

4.90641E-23

38.46786372

chr2_238213149_238213414

chr2

0

-

1.58339E-06

5.42033E-06

37.58090232

chr9_88891303_88891317

chr9

1

ISCA1

1.08415E-06

3.8617E-06

37.31409719

chr7_134279040_134279370

chr7

0

-

7.42127E-19

1.90865E-17

36.8404498

chr11_125477842_125477944

chr11

1

STT3A

1.37342E-52

3.02928E-50

36.14363165

chr4_85873367_85873551

chr4

1

WDFY3

8.771E-29

5.54172E-27

35.86458464

chr1_228243499_228243750

chr1

1

WNT3A

7.15126E-21

2.28614E-19

35.584194

chr2_131046231_131046636

chr2

5

MTND4P2|MTND5P2|MTND6P|AC068137.1|AC068137.1

1.32673E-40

1.75348E-38

35.46103678

chr7_39723763_39723940

chr7

1

RALA

2.01448E-11

1.85339E-10

34.0085943

chr6_132297421_132297621

chr6

1

RP11-69I8.

0.001384883

0.002169711

33.37310303

chr3_108880309_108880502

chr3

0

-

7.64309E-11

6.36725E-10

33.06601128

chr1_66753890_66754101

chr1

1

PDE4B

0.00182074

0.00275054

32.96104032

chr2_66654487_66654810

chr2

2

MEIS1-AS3|MEIS1

2.10022E-69

7.72486E-67

32.72071839

chr5_97764439_97764614

chr5

0

-

0.000291866

0.000557559

32.43193392

chr9_115957266_115957508

chr9

1

FKBP15

1.69011E-15

2.87555E-14

32.29050602

chr15_50209373_50209588

chr15

1

ATP8B4

3.01371E-25

1.45926E-23

31.98261372

chr17_11608628_11608839

chr17

1

DNAH9

6.97091E-30

4.8207E-28

31.79815222

chr4_3040168_3040507

chr4

2

GRK4|RNU6-204

7.3598E-14

9.96918E-13

31.65690685

chr6_88956453_88956584

chr6

0

-

9.99041E-12

9.68676E-11

31.54201058

chr21_15436054_15437354

chr21

3

AP001347.|ANKRD20A18|RNA5SP48

0

0

31.16688815

chr3_49712243_49712522

chr3

4

BSN|APEH|MST1|AC099668.

8.07314E-25

3.79115E-23

31.03180163

chr12_25264152_25264371

chr12

2

LRMP|CASC1

8.78331E-27

4.8313E-25

30.29058703

chrX_9121569_9121778

chrX

1

FAM9B

0.000426217

0.000777831

30.08088109

chr3_84932517_84932772

chr3

2

LINC00971|LINC02025

2.07283E-18

5.07715E-17

29.94169954

chr7_123928236_123928461

chr7

2

RP5-921G16.|RP11-264K23.

3.518E-31

2.67311E-29

29.91216787

chr9_106087270_106087465

chr9

1

LINC01492

8.36825E-19

2.13729E-17

29.78985686

chr5_120388958_120389191

chr5

2

AC008565.|CTD-2613O8.

7.61105E-44

1.17667E-41

29.58350371

chr11_8927411_8927650

chr11

2

ST5|AKIP1

4.49366E-10

3.25802E-09

29.52623011

chr6_159572123_159572413

chr6

0

-

7.42108E-39

8.88692E-37

29.31934038

chr22_49625975_49626187

chr22

0

-

2.92003E-11

2.61571E-10

29.27879223

chr12_9776968_9777167

chr12

2

LOC374443|RNU6-700

1.12651E-09

7.58502E-09

29.08923252

chr3_170332310_170332590

chr3

1

SLC7A14-AS1

1.8717E-08

9.81278E-08

29.01098901

chr4_77986307_77986536

chr4

1

CCNI

3.91084E-14

5.48555E-13

28.97085991

chr22_42176013_42176129

chr22

1

MEI1

0.000206038

0.000410706

28.81962113

chr10_25240859_25240994

chr10

2

PRTFDC1|RP11-165A20.

6.26124E-28

3.71906E-26

28.60587752

chr3_79966821_79966915

chr3

0

-

1.15638E-10

9.34284E-10

28.60217066

chr3_45649141_45649463

chr3

1

LIMD1

6.28165E-62

1.8899E-59

28.50314686

chr6_25166613_25166879

chr6

1

CMAHP

2.17351E-96

1.46968E-93

28.37986243

chr3_152213629_152213846

chr3

1

RP11-362A9.

6.8342E-05

0.000155335

28.26746276

chr11_59836664_59836958

chr11

2

MS4A3|RP11-736I10.

3.63343E-09

2.21257E-08

28.0516934

chr20_14124320_14124383

chr20

1

MACROD2

1.40341E-20

4.35847E-19

28.02413196

chr22_32728242_32728474

chr22

1

RP1-149A16.1

8.85421E-11

7.29047E-10

27.91646442

chr2_183106034_183106226

chr2

1

PDE1A

9.65506E-09

5.3791E-08

27.85254866

chr6_118401762_118401938

chr6

2

SLC35F1|LOC105377967

4.64459E-19

1.22408E-17

27.79487179

chr10_91369814_91369992

chr10

1

PANK1

1.57036E-21

5.3785E-20

27.77056277

chr12_49121182_49121257

chr12

1

TEX49

8.24722E-13

9.54004E-12

27.60425962

chr5_95362159_95362388

chr5

1

LOC101929710

4.61645E-19

1.21738E-17

27.51574796

chr17_16935456_16935522

chr17

0

-

4.58885E-09

2.74027E-08

27.46293683

chr3_42093299_42093629

chr3

2

TRAK1|RP11-193I22.

6.20321E-37

6.69223E-35

27.41943731

chr3_149867507_149867632

chr3

1

LOC105374313

3.07268E-11

2.74327E-10

27.29693742

chrX_43503885_43504133

chrX

0

-

4.40393E-09

2.64009E-08

27.18926273

chr14_77035752_77035960

chr14

1

RP11-187O7.

6.2288E-12

6.23127E-11

27.15821169

chr10_52771069_52771259

chr10

1

PRKG1

3.87096E-16

7.14936E-15

27.04932754

chr2_38055293_38055466

chr2

1

LINC00211

7.48568E-28

4.4228E-26

27.03032591

chr1_108245511_108245769

chr1

1

VAV3

3.17027E-12

3.3165E-11

26.94594044

chr11_132891631_132891819

chr11

1

OPCML

4.6077E-05

0.000109674

26.80024478

chr8_8538073_8538307

chr8

0

-

6.60262E-14

8.99085E-13

26.64249158

chr8_107757803_107757980

chr8

1

OXR1

0.000716855

0.001224528

26.62186488

chr21_36250641_36250940

chr21

2

LOC100506403|RUNX1

3.11176E-25

1.50565E-23

26.55673748

chr9_14910433_14910528

chr9

1

FREM1

8.33159E-21

2.64892E-19

26.53151371

chr3_10851357_10851586

chr3

1

SLC6A11

0.004410223

0.005890646

26.40737551

chr9_73216788_73217020

chr9

1

TRPM3

3.0921E-15

5.06212E-14

26.40133464

chr17_77906666_77906803

chr17

3

LINC01979|LINC01978|TBC1D16

3.90756E-08

1.9197E-07

26.38634157

chr15_101507155_101507328

chr15

1

LRRK1

5.16927E-09

3.05155E-08

26.37814064

chr10_87988149_87988359

chr10

1

GRID1

0.003497861

0.004828383

26.35618749

chr13_46978725_46978934

chr13

2

RUBCNL|RNU6-68

2.29746E-12

2.46329E-11

26.04182226

chrX_149197068_149197285

chrX

1

LINC00894

2.44211E-07

1.01031E-06

26.00779648

chr16_66453623_66453908

chr16

2

LINC00920|BEAN1

3.68957E-06

1.15579E-05

25.85630262

chr1_165186330_165186523

chr1

3

LMX1A|RP11-38C18.|RP11-38C18.

0.000275984

0.000530947

25.72822521

chr12_6662473_6662654

chr12

3

IFFO1|RP5-940J5.|NOP2

9.5204E-29

6.00387E-27

25.71991419

chr4_129308317_129308548

chr4

1

LINC02615

2.02201E-09

1.29361E-08

25.68972744

chr4_102756899_102757268

chr4

1

BANK1

8.27941E-46

1.39081E-43

25.66642434

chr6_101852800_101852969

chr6

1

GRIK2

1.14283E-19

3.23959E-18

25.50742035

chr7_147500652_147500831

chr7

1

CNTNAP2

4.69478E-06

1.43535E-05

25.47112462

chr2_130683180_130683508

chr2

2

PLAC9P1|LINC01856

2.6716E-16

5.03565E-15

25.46823547

chr3_69157750_69157970

chr3

2

ARL6IP5|LMOD3

6.39788E-06

1.89017E-05

25.46542143

chr12_104676677_104676883

chr12

2

TXNRD1|RP11-818F20.

4.32249E-15

6.92567E-14

25.4273867

chr9_4375797_4376098

chr9

1

AL162419.

3.07166E-06

9.81114E-06

25.41113806

chr6_55377549_55377764

chr6

1

HMGCLL1

4.38355E-45

7.21874E-43

25.36445879

chr8_80816076_80816210

chr8

0

-

2.50289E-06

8.16528E-06

25.34385329

chr5_171810643_171810847

chr5

1

SH3PXD2B

2.88223E-10

2.16508E-09

25.2488391

chr2_66648614_66648933

chr2

1

MEIS1-AS3

1.28929E-22

4.90242E-21

25.21383356

chr14_96583490_96583604

chr14

0

-

0.003078364

0.004325989

25.19935377

chr22_16864741_16864893

chr22

1

ABCD1P

0.002259786

0.003313591

25.19298246

chr10_90483503_90483709

chr10

2

LIPK|KRT8P3

3.47526E-44

5.45461E-42

25.15294597

chr6_7699982_7700169

chr6

0

-

1.47903E-12

1.64125E-11

25.12462285

chr3_195501977_195502276

chr3

1

MUC4

1.31243E-47

2.3649E-45

25.08813062

chr18_29303346_29303538

chr18

3

RN7SKP4|LRRC37A7|RP11-549B18.

1.62011E-27

9.3655E-26

25.03570419

chr3_95424692_95425326

chr3

0

-

3.3968E-14

4.80683E-13

25.03516155

chr3_180102299_180102384

chr3

0

-

9.13491E-07

3.3066E-06

25.01885857

chr22_35587422_35587713

chr22

2

LINC01399|COX7BP

1.08406E-10

8.79565E-10

-25.01400968

chr20_36132841_36133069

chr20

1

BLCAP

1.55091E-18

3.84306E-17

-25.14666861

chr19_47082930_47083060

chr19

2

PPP5D1|AC011551.

6.29188E-21

2.02349E-19

-25.24712874

chr19_49572906_49573157

chr19

4

NTF4|CTB-60B18.1|CTB-60B18.1|KCNA7

5.89203E-07

2.23171E-06

-25.34019384

chr3_11489044_11489192

chr3

1

ATG7

7.90562E-10

5.47842E-09

-25.40198715

chr17_75631717_75631979

chr17

0

-

1.84137E-05

4.85349E-05

-25.53803605

chr1_19176711_19177105

chr1

2

TAS1R2|RP13-279N23.

0.000375134

0.000695164

-25.58525242

chr11_5840963_5841191

chr11

2

TRIM5|OR52N2

2.88714E-66

9.68772E-64

-25.69185678

chr2_55378165_55378300

chr2

0

-

9.89458E-19

2.51065E-17

-25.78431373

chr13_19315890_19316198

chr13

3

ZNF965|LINC00417|CYP4F34

8.90044E-14

1.19079E-12

-25.87364383

chr18_61669993_61670289

chr18

1

SERPINB8

8.41432E-63

2.5838E-60

-26.04262437

chr20_32885922_32886673

chr20

1

AHCY

2.8332E-44

4.47838E-42

-26.45197851

chr6_106252261_106252558

chr6

0

-

6.2211E-107

4.9875E-104

-26.54101932

chr2_25383849_25384399

chr2

3

EFR3B|RP11-509E16|POMC

4.25503E-85

2.26062E-82

-26.70759651

chr4_131633369_131633569

chr4

0

-

1.21778E-09

8.14225E-09

-26.8218303

chr4_26789871_26790007

chr4

0

-

9.76527E-06

2.75795E-05

-27.15229384

chr1_55370080_55370265

chr1

0

-

1.33489E-60

3.89366E-58

-27.2302102

chr19_45619267_45619497

chr19

2

MARK4|PPP1R37

3.61011E-48

6.62101E-46

-27.29034787

chr16_33588844_33589010

chr16

1

ENPP7P13

8.03476E-26

4.0933E-24

-27.38040136

chr4_15907926_15908145

chr4

0

-

2.8442E-50

5.76083E-48

-27.44818039

chr10_67155111_67155236

chr10

0

-

4.83108E-11

4.17264E-10

-27.80333564

chr17_4278292_4278786

chr17

1

UBE2G1

9.00939E-24

3.82074E-22

-27.82640422

chr11_116344108_116344147

chr11

0

-

0.000326633

0.000615342

-27.8388828

chr19_34245878_34246069

chr19

1

CHST8

6.69284E-06

1.96741E-05

-27.92649973

chr17_16864290_16864524

chr17

1

TNFRSF13B

1.26001E-12

1.41214E-11

-28.2842299

chr11_82354426_82354705

chr11

2

MIR4300HG|RP11-179A16.

6.09835E-07

2.30172E-06

-29.18358119

chr5_153676389_153676633

chr5

1

GALNT10

1.43406E-68

5.14737E-66

-29.70725527

chr10_134778418_134779164

chr10

3

LINC01166|LINC01167|LINC01168

5.1864E-178

1.2399E-174

-29.73911375

chr8_36995835_36995986

chr8

1

MIR1268A

3.40507E-09

2.0846E-08

-29.75107115

chr2_213316451_213316721

chr2

1

ERBB4

3.25312E-05

8.05895E-05

-30.06050559

chr3_105601223_105601548

chr3

0

-

3.99011E-26

2.08106E-24

-30.45696192

chr2_130693963_130694229

chr2

3

PLAC9P1|LINC01856|AC079776.

1.8989E-114

1.6726E-111

-30.54653298

chr11_110986763_110986975

chr11

1

RP11-89C3.

0

0

-30.61868687

chr4_184296182_184296485

chr4

2

RP11-451F20.|AC093844.

3.2812E-82

1.64531E-79

-30.6235367

chr6_21452696_21452935

chr6

0

-

4.66098E-09

2.77827E-08

-30.7652865

chr14_56669107_56669307

chr14

1

PELI2

0.005344129

0.006946025

-31.10639369

chr13_25141770_25142091

chr13

3

TPTE2P6|PSPC1P|RP11-556N21.

2.11697E-25

1.04092E-23

-31.10709988

chr2_238796929_238797268

chr2

1

RAMP1

9.04001E-24

3.83251E-22

-31.35072812

chr17_6901611_6901751

chr17

4

ALOX12-AS1|RP11-589P10.|RP11-589P10.|ALOX12

0.00014541

0.000302279

-31.60823695

chr15_79299672_79299707

chr15

1

RASGRF1

0.004476538

0.005967375

-32.72230999

chr1_223353214_223353363

chr1

1

RP11-239E10

7.13359E-18

1.64468E-16

-33.00889209

chr1_113221602_113221746

chr1

2

CAPZA1|MOV10

8.44636E-34

7.62526E-32

-33.03827068

chr19_21860552_21861267

chr19

1

RP11-420K14.

1.3469E-284

1.127E-280

-33.08420719

chr2_145505959_145506291

chr2

2

TEX41|AC023128.

2.36911E-23

9.63501E-22

-33.13077409

chr21_29565178_29565285

chr21

1

LINC01695

0.000196292

0.000393653

-33.4915241

chr22_47442504_47442671

chr22

1

TBC1D22A

2.44869E-31

1.87658E-29

-34.87001855

chr8_119449323_119449547

chr8

1

SAMD12

2.1869E-06

7.23312E-06

-36.3138769

chr3_6706652_6706860

chr3

2

AC069277.|GRM7-AS3

0.00028562

0.000547251

-39.63810451

chr8_129286309_129286584

chr8

0

-

3.30084E-09

2.02608E-08

-39.75752508

chr12_64067778_64067878

chr12

3

MIR548Z|DPY19L2|RP11-415I12.

0.000389652

0.000718873

-40.8170893

chr15_71187717_71187868

chr15

2

LRRC49|THAP10

0.004522921

0.006020138

-42.45762712

chr3_115099771_115100023

chr3

0

-

0.002555418

0.003683188

-42.60471914

chr17_57187586_57187924

chr17

3

TRIM37|AC099850.|SKA2

2.39899E-60

6.93704E-58

-46.21784352

To identify the molecular pathways and functions potentially influenced by methylation changes in AN, we performed GO term and KEGG pathway enrichment analyses of the genes closest to the identified DMRs (within the gene body or within +/- 10kb of gene start/end sites) using the DAVID bioinformatics resources 6.8 (https://david.ncifcrf.gov/). When "Disease" was used for categorization, there were 5 charts categories (with a significant p value with Benjamini correction) of DMRs, the more significant charts being waist-hip ratio (n=9 counts, Benjamini 5.2e-3), tobacco use disorder (37 counts, Benjamini 2.8e-3), chemdependency (n=40 counts, Benjamini 1.2e-3), metabolic (n=49, Benjamini 3.1e-3) and neurological (32 counts, Benjamini 3.8e-3).

We next focused on the 153 DMRs, the String network analysis software identified a subnetwork, related to MAPK signaling pathway (hs04010, FDR 0.0026), PI3K-Akt signaling pathway (hsa04151, FDR 0.0232) and neurotrophin signaling pathway (hsa04550, FDR 0.0251)(https://string-db.org/cgi/network.pl?taskId=07ri9Goe1SzI) (Figure 1).

Figure 1. Protein subnetwork of the identified proteins constructed with the STRING software for enrichment analysis of proteins (PPI enrichment value 7.04e-08) showing a dysmethylation status (15 DMRs among the 119 identified DMRs including at least one coding or non-coding genes). Each node represents a protein entity. Gene Ontology identified functional enrichments for several biological process including neuron differentiation (GO:0030182, FDR 8.15e-06) and behavior (GO:0007610, FDR 0.00025)(in black), and KEGG pathways identified enrichments for several pathways including MAPK signaling pathway  (hs04010, FDR 0.0026)(grey square), PI3K-Akt signaling pathway (hsa04151, FDR 0.0232)(white square) and Neurotrophin signaling pathway (hsa04550, FDR 0.0251) (grey triangle)(https://string-db.org/cgi/network.pl?taskId=07ri9Goe1SzI)

Discussion

To the best of our knowledge, the present study presents the first genome-wide DNA methylation profiling of 6 male AN patients, using a high-throughput DNA methylation sequencing covering a large number of CpG sites on the human genome. Patients affected with AN restrictive type showed many differentially methylated sites, with significant between group differences corresponding to genes implicated in metabolic and nutritional status, psychiatric status and immune function. When we compared our DMCs list with the short list of Steiger and colleagues obtained in female AN patients, 32 genes were found in both lists [9]. Moreover, because the interaction between depression and anorexia nervosa was significant, we also identified six genes previously found in patients with major depressive disorder [8]. Taking into account that our approach was based on genome-wide methylation sequencing, we also identified additional candidate genes involved in psychiatric disorders (GRID1, NAALADL2-AS3, PDZD, CEP85L, GRIK3, SLC7A14), in metabolism regulation (lipids, ZFP36L1, CERK, ACSF3, EEPD1; glucose, KCNA7), and finally, in addiction (GDE1, CCKAR, PDYN). Moreover, we identified few genes previously associated with anorexia nervosa in genome-wide association studies (VGLL4, GRID1, WWOX, CAMK1D, SORCS2) (https://www.ebi.ac.uk/gwas/search?query=anorexia%20nervosa). Interestingly, the String network analysis software revealed a subnetwork related to neurotrophin signaling pathway (Figure 1). The reported data, in addition to the previous reported findings for BDNF, NTRK2 and NTRK3, point again neurotrophin signaling genes as key regulators of eating behavior.

To conclude, our data replicates several results concerning several target genes such as PRKAG2, RPTOR, and ICAM5 previously discussed [9], and identified novel signaling pathways involving PI3K-Akt and neurotrophin signaling pathway disturbed in anorexia nervosa. Future replication of findings in male AN patients will be a determinant.

Compliance with ethical standards

Funding

This study was supported by Institut National de la Recherche Médicale (INSERM) and by Fondation Maladies Rares (program High throughput sequencing and rare disease, number #11632).

Conflicts of interest

The authors declare that they have no conflicts of interest.

Ethical approval

All procédures performed in this study were in accordance with the ethical standards of our national research committee and with the 1964 Helsinki declaration and its later amendments. Informed written consent was obtained from all patients and parents included in the study.

Acknowledgements

We thank the families for their enthusiastic participation and all physicians from the different medical and psychiatric centers.

View supplementary data

References

  1. Bulik CM, Sullivan PF, Tozzi F, Furberg H, Lichtenstein P, et al. (2006) Prevalence, heritability, and prospective risk factors for anorexia nervosa. Arch Gen Psychiatry 63: 305-312. [Crossref]
  2. Wang K, Zhang H, Bloss CS, Duvvuri V, Kaye W, Schork NJ, et al. (2011) A genome-wide association study on common SNPs and rare CNVs in anorexia nervosa. Mol Psychiatry 16: 949-959. [Crossref]
  3. Boraska V, Franklin CS, Floyd JA, Thornton LM, Huckins LM, et al. (2014) A genome-wide association study of anorexia nervosa. Mol Psychiatry 19: 1085-1094. [Crossref]
  4. Duncan L, Yilmaz Z, Gaspar H, Walters R, GoldsteinJ, Anttila V, et al. (2017) Significant locus and metabolic genetic correlations revealed in genome-wide association study of anorexia nervosa. Am J Psychiatry 174: 850-858. [Crossref]
  5. Huckins LM, Hatzikotoulas K, Southam L, Thornton LM, Steinberg J, Aguilera-McKay F, et al. (2018) Revealing the complex genetic architecture of obsessive-compulsive disorder using meta-analysis. Mol Psychiatry 23: 1181-1188. [Crossref]
  6. Thornton LM, Welch E, Munn-Chernoff MA, Lichtenstein P, Bulik CM (2016) Anorexia nervosa, major depression, and suicide attempts: shared genetic factors. Suicide Life Threat Behav 46: 525-534. [Crossref]
  7. Wade TD, Bulik CM, Neale M, Kendler KS (2000) Anorexia nervosa and major depression: shared genetic and environmental risk factors. Am J Psychiatry 157: 469-471. [Crossref]
  8. Shimada M, Otowa T, Miyagawa T, Umekage T, Kawamura Y, Bundo M, et al. (2018) Anepigenome-wide methylation study of healthy individuals with or without depressive symptoms. J Hum Genet 63: 319-326. [Crossref]
  9. Steiger H, Booij L, Kahan `, McGregor K, Thaler L, Fletcher E, et al. (2019) A longitudinal,epigenome-wide study of DNA methylation in anorexia nervosa: results in actively ill, partially weight-restored, long-term remitted and non-eating-disorderedwomen. J Psychiatry Neurosci 44: 1-9. [Crossref]
  10. Akalin A, Kormaksson M, Li S, Garrett-Bakelman FE, Figueroa ME, Melnick A, et al. (2012) MethylKit: a comprehensive R package for the analysis of genome-wideDNA methylation profiles. Genome Biol 13: 80-87. [Crossref]

Editorial Information

Editor-in-Chief

Article Type

Research Article

Publication history

Received date: July 12, 2019
Accepted date: July 22, 2019
Published date: July 30, 2019

Copyright

© 2019 Kim A. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

Citation

Kim A (2019) Altered DNA methylation associated with nervosa anorexia in males. Integr Mol Med 6 DOI: 10.15761/IMM.1000376

Corresponding author

Thierry Bienvenu

Institute of Psychiatry and Neuroscience of Paris, 102 rue de la Santé, 75014 Paris, France

E-mail : bhuvaneswari.bibleraaj@uhsm.nhs.uk

Figure 1. Protein subnetwork of the identified proteins constructed with the STRING software for enrichment analysis of proteins (PPI enrichment value 7.04e-08) showing a dysmethylation status (15 DMRs among the 119 identified DMRs including at least one coding or non-coding genes). Each node represents a protein entity. Gene Ontology identified functional enrichments for several biological process including neuron differentiation (GO:0030182, FDR 8.15e-06) and behavior (GO:0007610, FDR 0.00025)(in black), and KEGG pathways identified enrichments for several pathways including MAPK signaling pathway  (hs04010, FDR 0.0026)(grey square), PI3K-Akt signaling pathway (hsa04151, FDR 0.0232)(white square) and Neurotrophin signaling pathway (hsa04550, FDR 0.0251) (grey triangle)(https://string-db.org/cgi/network.pl?taskId=07ri9Goe1SzI)

Table 1. Identification of differentially methylated regions (DMR) was performed using the predefined regions of Illumina TruSeq Methylcapture EPIC kit presenting at least 25 % methylation difference between ANcases and controls (column Change) with a q-value (FDR corrected p-value) threshold of 0.01

DMR

Chr

Nb Genes

Genes

p value

q value

Change

chrX_114502896_114503051

chrX

0

-

2.22451E-15

3.71584E-14

59.78484933

chr14_54375093_54375328

chr14

1

AL138479.

2.65207E-24

1.18277E-22

48.00682879

chr8_80036078_80036321

chr8

0

-

2.44128E-36

2.55548E-34

46.08190008

chr11_79289117_79289197

chr11

0

-

2.09678E-48

3.89352E-46

44.01889299

chr2_165342265_165342368

chr2

1

GRB14

5.17941E-11

4.44995E-10

43.62139918

chr5_118811413_118811668

chr5

2

HSD17B4|snoU1

4.63731E-11

4.01928E-10

43.59922516

chr19_45113900_45114110

chr19

2

CEACAM22P|IGSF23

9.32156E-32

7.39337E-30

40.63822929

chr12_39885082_39885307

chr12

0

-

1.48466E-54

3.57503E-52

40.49521423

chr11_99564881_99564954

chr11

1

CNTN5

9.37129E-15

1.42916E-13

39.62334329

chr20_46228572_46228783

chr20

1

NCOA3

1.11712E-13

1.47266E-12

39.56217898

chr12_9507242_9507307

chr12

0

-

1.05726E-24

4.90641E-23

38.46786372

chr2_238213149_238213414

chr2

0

-

1.58339E-06

5.42033E-06

37.58090232

chr9_88891303_88891317

chr9

1

ISCA1

1.08415E-06

3.8617E-06

37.31409719

chr7_134279040_134279370

chr7

0

-

7.42127E-19

1.90865E-17

36.8404498

chr11_125477842_125477944

chr11

1

STT3A

1.37342E-52

3.02928E-50

36.14363165

chr4_85873367_85873551

chr4

1

WDFY3

8.771E-29

5.54172E-27

35.86458464

chr1_228243499_228243750

chr1

1

WNT3A

7.15126E-21

2.28614E-19

35.584194

chr2_131046231_131046636

chr2

5

MTND4P2|MTND5P2|MTND6P|AC068137.1|AC068137.1

1.32673E-40

1.75348E-38

35.46103678

chr7_39723763_39723940

chr7

1

RALA

2.01448E-11

1.85339E-10

34.0085943

chr6_132297421_132297621

chr6

1

RP11-69I8.

0.001384883

0.002169711

33.37310303

chr3_108880309_108880502

chr3

0

-

7.64309E-11

6.36725E-10

33.06601128

chr1_66753890_66754101

chr1

1

PDE4B

0.00182074

0.00275054

32.96104032

chr2_66654487_66654810

chr2

2

MEIS1-AS3|MEIS1

2.10022E-69

7.72486E-67

32.72071839

chr5_97764439_97764614

chr5

0

-

0.000291866

0.000557559

32.43193392

chr9_115957266_115957508

chr9

1

FKBP15

1.69011E-15

2.87555E-14

32.29050602

chr15_50209373_50209588

chr15

1

ATP8B4

3.01371E-25

1.45926E-23

31.98261372

chr17_11608628_11608839

chr17

1

DNAH9

6.97091E-30

4.8207E-28

31.79815222

chr4_3040168_3040507

chr4

2

GRK4|RNU6-204

7.3598E-14

9.96918E-13

31.65690685

chr6_88956453_88956584

chr6

0

-

9.99041E-12

9.68676E-11

31.54201058

chr21_15436054_15437354

chr21

3

AP001347.|ANKRD20A18|RNA5SP48

0

0

31.16688815

chr3_49712243_49712522

chr3

4

BSN|APEH|MST1|AC099668.

8.07314E-25

3.79115E-23

31.03180163

chr12_25264152_25264371

chr12

2

LRMP|CASC1

8.78331E-27

4.8313E-25

30.29058703

chrX_9121569_9121778

chrX

1

FAM9B

0.000426217

0.000777831

30.08088109

chr3_84932517_84932772

chr3

2

LINC00971|LINC02025

2.07283E-18

5.07715E-17

29.94169954

chr7_123928236_123928461

chr7

2

RP5-921G16.|RP11-264K23.

3.518E-31

2.67311E-29

29.91216787

chr9_106087270_106087465

chr9

1

LINC01492

8.36825E-19

2.13729E-17

29.78985686

chr5_120388958_120389191

chr5

2

AC008565.|CTD-2613O8.

7.61105E-44

1.17667E-41

29.58350371

chr11_8927411_8927650

chr11

2

ST5|AKIP1

4.49366E-10

3.25802E-09

29.52623011

chr6_159572123_159572413

chr6

0

-

7.42108E-39

8.88692E-37

29.31934038

chr22_49625975_49626187

chr22

0

-

2.92003E-11

2.61571E-10

29.27879223

chr12_9776968_9777167

chr12

2

LOC374443|RNU6-700

1.12651E-09

7.58502E-09

29.08923252

chr3_170332310_170332590

chr3

1

SLC7A14-AS1

1.8717E-08

9.81278E-08

29.01098901

chr4_77986307_77986536

chr4

1

CCNI

3.91084E-14

5.48555E-13

28.97085991

chr22_42176013_42176129

chr22

1

MEI1

0.000206038

0.000410706

28.81962113

chr10_25240859_25240994

chr10

2

PRTFDC1|RP11-165A20.

6.26124E-28

3.71906E-26

28.60587752

chr3_79966821_79966915

chr3

0

-

1.15638E-10

9.34284E-10

28.60217066

chr3_45649141_45649463

chr3

1

LIMD1

6.28165E-62

1.8899E-59

28.50314686

chr6_25166613_25166879

chr6

1

CMAHP

2.17351E-96

1.46968E-93

28.37986243

chr3_152213629_152213846

chr3

1

RP11-362A9.

6.8342E-05

0.000155335

28.26746276

chr11_59836664_59836958

chr11

2

MS4A3|RP11-736I10.

3.63343E-09

2.21257E-08

28.0516934

chr20_14124320_14124383

chr20

1

MACROD2

1.40341E-20

4.35847E-19

28.02413196

chr22_32728242_32728474

chr22

1

RP1-149A16.1

8.85421E-11

7.29047E-10

27.91646442

chr2_183106034_183106226

chr2

1

PDE1A

9.65506E-09

5.3791E-08

27.85254866

chr6_118401762_118401938

chr6

2

SLC35F1|LOC105377967

4.64459E-19

1.22408E-17

27.79487179

chr10_91369814_91369992

chr10

1

PANK1

1.57036E-21

5.3785E-20

27.77056277

chr12_49121182_49121257

chr12

1

TEX49

8.24722E-13

9.54004E-12

27.60425962

chr5_95362159_95362388

chr5

1

LOC101929710

4.61645E-19

1.21738E-17

27.51574796

chr17_16935456_16935522

chr17

0

-

4.58885E-09

2.74027E-08

27.46293683

chr3_42093299_42093629

chr3

2

TRAK1|RP11-193I22.

6.20321E-37

6.69223E-35

27.41943731

chr3_149867507_149867632

chr3

1

LOC105374313

3.07268E-11

2.74327E-10

27.29693742

chrX_43503885_43504133

chrX

0

-

4.40393E-09

2.64009E-08

27.18926273

chr14_77035752_77035960

chr14

1

RP11-187O7.

6.2288E-12

6.23127E-11

27.15821169

chr10_52771069_52771259

chr10

1

PRKG1

3.87096E-16

7.14936E-15

27.04932754

chr2_38055293_38055466

chr2

1

LINC00211

7.48568E-28

4.4228E-26

27.03032591

chr1_108245511_108245769

chr1

1

VAV3

3.17027E-12

3.3165E-11

26.94594044

chr11_132891631_132891819

chr11

1

OPCML

4.6077E-05

0.000109674

26.80024478

chr8_8538073_8538307

chr8

0

-

6.60262E-14

8.99085E-13

26.64249158

chr8_107757803_107757980

chr8

1

OXR1

0.000716855

0.001224528

26.62186488

chr21_36250641_36250940

chr21

2

LOC100506403|RUNX1

3.11176E-25

1.50565E-23

26.55673748

chr9_14910433_14910528

chr9

1

FREM1

8.33159E-21

2.64892E-19

26.53151371

chr3_10851357_10851586

chr3

1

SLC6A11

0.004410223

0.005890646

26.40737551

chr9_73216788_73217020

chr9

1

TRPM3

3.0921E-15

5.06212E-14

26.40133464

chr17_77906666_77906803

chr17

3

LINC01979|LINC01978|TBC1D16

3.90756E-08

1.9197E-07

26.38634157

chr15_101507155_101507328

chr15

1

LRRK1

5.16927E-09

3.05155E-08

26.37814064

chr10_87988149_87988359

chr10

1

GRID1

0.003497861

0.004828383

26.35618749

chr13_46978725_46978934

chr13

2

RUBCNL|RNU6-68

2.29746E-12

2.46329E-11

26.04182226

chrX_149197068_149197285

chrX

1

LINC00894

2.44211E-07

1.01031E-06

26.00779648

chr16_66453623_66453908

chr16

2

LINC00920|BEAN1

3.68957E-06

1.15579E-05

25.85630262

chr1_165186330_165186523

chr1

3

LMX1A|RP11-38C18.|RP11-38C18.

0.000275984

0.000530947

25.72822521

chr12_6662473_6662654

chr12

3

IFFO1|RP5-940J5.|NOP2

9.5204E-29

6.00387E-27

25.71991419

chr4_129308317_129308548

chr4

1

LINC02615

2.02201E-09

1.29361E-08

25.68972744

chr4_102756899_102757268

chr4

1

BANK1

8.27941E-46

1.39081E-43

25.66642434

chr6_101852800_101852969

chr6

1

GRIK2

1.14283E-19

3.23959E-18

25.50742035

chr7_147500652_147500831

chr7

1

CNTNAP2

4.69478E-06

1.43535E-05

25.47112462

chr2_130683180_130683508

chr2

2

PLAC9P1|LINC01856

2.6716E-16

5.03565E-15

25.46823547

chr3_69157750_69157970

chr3

2

ARL6IP5|LMOD3

6.39788E-06

1.89017E-05

25.46542143

chr12_104676677_104676883

chr12

2

TXNRD1|RP11-818F20.

4.32249E-15

6.92567E-14

25.4273867

chr9_4375797_4376098

chr9

1

AL162419.

3.07166E-06

9.81114E-06

25.41113806

chr6_55377549_55377764

chr6

1

HMGCLL1

4.38355E-45

7.21874E-43

25.36445879

chr8_80816076_80816210

chr8

0

-

2.50289E-06

8.16528E-06

25.34385329

chr5_171810643_171810847

chr5

1

SH3PXD2B

2.88223E-10

2.16508E-09

25.2488391

chr2_66648614_66648933

chr2

1

MEIS1-AS3

1.28929E-22

4.90242E-21

25.21383356

chr14_96583490_96583604

chr14

0

-

0.003078364

0.004325989

25.19935377

chr22_16864741_16864893

chr22

1

ABCD1P

0.002259786

0.003313591

25.19298246

chr10_90483503_90483709

chr10

2

LIPK|KRT8P3

3.47526E-44

5.45461E-42

25.15294597

chr6_7699982_7700169

chr6

0

-

1.47903E-12

1.64125E-11

25.12462285

chr3_195501977_195502276

chr3

1

MUC4

1.31243E-47

2.3649E-45

25.08813062

chr18_29303346_29303538

chr18

3

RN7SKP4|LRRC37A7|RP11-549B18.

1.62011E-27

9.3655E-26

25.03570419

chr3_95424692_95425326

chr3

0

-

3.3968E-14

4.80683E-13

25.03516155

chr3_180102299_180102384

chr3

0

-

9.13491E-07

3.3066E-06

25.01885857

chr22_35587422_35587713

chr22

2

LINC01399|COX7BP

1.08406E-10

8.79565E-10

-25.01400968

chr20_36132841_36133069

chr20

1

BLCAP

1.55091E-18

3.84306E-17

-25.14666861

chr19_47082930_47083060

chr19

2

PPP5D1|AC011551.

6.29188E-21

2.02349E-19

-25.24712874

chr19_49572906_49573157

chr19

4

NTF4|CTB-60B18.1|CTB-60B18.1|KCNA7

5.89203E-07

2.23171E-06

-25.34019384

chr3_11489044_11489192

chr3

1

ATG7

7.90562E-10

5.47842E-09

-25.40198715

chr17_75631717_75631979

chr17

0

-

1.84137E-05

4.85349E-05

-25.53803605

chr1_19176711_19177105

chr1

2

TAS1R2|RP13-279N23.

0.000375134

0.000695164

-25.58525242

chr11_5840963_5841191

chr11

2

TRIM5|OR52N2

2.88714E-66

9.68772E-64

-25.69185678

chr2_55378165_55378300

chr2

0

-

9.89458E-19

2.51065E-17

-25.78431373

chr13_19315890_19316198

chr13

3

ZNF965|LINC00417|CYP4F34

8.90044E-14

1.19079E-12

-25.87364383

chr18_61669993_61670289

chr18

1

SERPINB8

8.41432E-63

2.5838E-60

-26.04262437

chr20_32885922_32886673

chr20

1

AHCY

2.8332E-44

4.47838E-42

-26.45197851

chr6_106252261_106252558

chr6

0

-

6.2211E-107

4.9875E-104

-26.54101932

chr2_25383849_25384399

chr2

3

EFR3B|RP11-509E16|POMC

4.25503E-85

2.26062E-82

-26.70759651

chr4_131633369_131633569

chr4

0

-

1.21778E-09

8.14225E-09

-26.8218303

chr4_26789871_26790007

chr4

0

-

9.76527E-06

2.75795E-05

-27.15229384

chr1_55370080_55370265

chr1

0

-

1.33489E-60

3.89366E-58

-27.2302102

chr19_45619267_45619497

chr19

2

MARK4|PPP1R37

3.61011E-48

6.62101E-46

-27.29034787

chr16_33588844_33589010

chr16

1

ENPP7P13

8.03476E-26

4.0933E-24

-27.38040136

chr4_15907926_15908145

chr4

0

-

2.8442E-50

5.76083E-48

-27.44818039

chr10_67155111_67155236

chr10

0

-

4.83108E-11

4.17264E-10

-27.80333564

chr17_4278292_4278786

chr17

1

UBE2G1

9.00939E-24

3.82074E-22

-27.82640422

chr11_116344108_116344147

chr11

0

-

0.000326633

0.000615342

-27.8388828

chr19_34245878_34246069

chr19

1

CHST8

6.69284E-06

1.96741E-05

-27.92649973

chr17_16864290_16864524

chr17

1

TNFRSF13B

1.26001E-12

1.41214E-11

-28.2842299

chr11_82354426_82354705

chr11

2

MIR4300HG|RP11-179A16.

6.09835E-07

2.30172E-06

-29.18358119

chr5_153676389_153676633

chr5

1

GALNT10

1.43406E-68

5.14737E-66

-29.70725527

chr10_134778418_134779164

chr10

3

LINC01166|LINC01167|LINC01168

5.1864E-178

1.2399E-174

-29.73911375

chr8_36995835_36995986

chr8

1

MIR1268A

3.40507E-09

2.0846E-08

-29.75107115

chr2_213316451_213316721

chr2

1

ERBB4

3.25312E-05

8.05895E-05

-30.06050559

chr3_105601223_105601548

chr3

0

-

3.99011E-26

2.08106E-24

-30.45696192

chr2_130693963_130694229

chr2

3

PLAC9P1|LINC01856|AC079776.

1.8989E-114

1.6726E-111

-30.54653298

chr11_110986763_110986975

chr11

1

RP11-89C3.

0

0

-30.61868687

chr4_184296182_184296485

chr4

2

RP11-451F20.|AC093844.

3.2812E-82

1.64531E-79

-30.6235367

chr6_21452696_21452935

chr6

0

-

4.66098E-09

2.77827E-08

-30.7652865

chr14_56669107_56669307

chr14

1

PELI2

0.005344129

0.006946025

-31.10639369

chr13_25141770_25142091

chr13

3

TPTE2P6|PSPC1P|RP11-556N21.

2.11697E-25

1.04092E-23

-31.10709988

chr2_238796929_238797268

chr2

1

RAMP1

9.04001E-24

3.83251E-22

-31.35072812

chr17_6901611_6901751

chr17

4

ALOX12-AS1|RP11-589P10.|RP11-589P10.|ALOX12

0.00014541

0.000302279

-31.60823695

chr15_79299672_79299707

chr15

1

RASGRF1

0.004476538

0.005967375

-32.72230999

chr1_223353214_223353363

chr1

1

RP11-239E10

7.13359E-18

1.64468E-16

-33.00889209

chr1_113221602_113221746

chr1

2

CAPZA1|MOV10

8.44636E-34

7.62526E-32

-33.03827068

chr19_21860552_21861267

chr19

1

RP11-420K14.

1.3469E-284

1.127E-280

-33.08420719

chr2_145505959_145506291

chr2

2

TEX41|AC023128.

2.36911E-23

9.63501E-22

-33.13077409

chr21_29565178_29565285

chr21

1

LINC01695

0.000196292

0.000393653

-33.4915241

chr22_47442504_47442671

chr22

1

TBC1D22A

2.44869E-31

1.87658E-29

-34.87001855

chr8_119449323_119449547

chr8

1

SAMD12

2.1869E-06

7.23312E-06

-36.3138769

chr3_6706652_6706860

chr3

2

AC069277.|GRM7-AS3

0.00028562

0.000547251

-39.63810451

chr8_129286309_129286584

chr8

0

-

3.30084E-09

2.02608E-08

-39.75752508

chr12_64067778_64067878

chr12

3

MIR548Z|DPY19L2|RP11-415I12.

0.000389652

0.000718873

-40.8170893

chr15_71187717_71187868

chr15

2

LRRC49|THAP10

0.004522921

0.006020138

-42.45762712

chr3_115099771_115100023

chr3

0

-

0.002555418

0.003683188

-42.60471914

chr17_57187586_57187924

chr17

3

TRIM37|AC099850.|SKA2

2.39899E-60

6.93704E-58

-46.21784352