carboxyl-ester lipase maturity-onset diabetes of the young...

1

Carboxyl-ester lipase Maturity-Onset Diabetes of the

Young is Associated with Development of Pancreatic

Cysts and Upregulated MAPK Signaling in Secretin-

stimulated duodenal fluid

Helge Ræder 1,2,3, Fiona E. McAllister4*, Erling Tjora2,3*, Shweta Bhatt1, Ingfrid

Haldorsen5,6, Jiang Hu1, Stefan M. Willems4, Mette Vesterhus7, Abdelfattah El Ouaamari1,

Manway Liu8, Maria B. Raeder

3, Heike Immervoll

9,10,11, Dag Hoem

12, Georg Dimcevski

7 ,

Pål R. Njolstad2,3, Anders Molven

9,10, Steven P Gygi

4, Rohit N. Kulkarni

1

From the 1Section of Islet Cell Biology and Regenerative Medicine, Joslin Diabetes Center,

Harvard Medical School, Boston, MA, USA, 2Department of Pediatrics, Haukeland University

Hospital, Bergen, Norway; 3Department of Clinical Science, University of Bergen, Bergen,

Norway, 4Department of Cell Biology, Harvard Medical School, Boston, MA, USA,5Department

of Radiology, Haukeland University Hospital, Bergen, Norway; 6Section for Radiology,

Department of Clinical Medicine, University of Bergen, 7Department of Medicine, Haukeland

University Hospital, Bergen, Norway, 8Department of Biomedical Engineering, Boston

University, Boston, MA,USA , 9The Gade Laboratory for Pathology, Department of Clinical

Medicine, University of Bergen, Norway; 10Department of Pathology, Haukeland University

Hospital, Bergen, Norway, 11Department of Pathology, Ålesund Hospital, Ålesund, Norway;

12Department of Surgery, Haukeland University Hospital, Bergen, Norway, *,These authors

contributed equally.

Corresponding author: Helge Raeder, [email protected]

Page 2 of 61Diabetes

Diabetes Publish Ahead of Print, published online September 23, 2013

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Running title: PANCREATIC CYSTS AND MAPK SIGNALING IN CEL-MODY

Original article: Word count, abstract: 200, Word count, main text: 3161, Number of references:

33, Number of figures: 5, Number of tables: 2.

ABSTRACT

CEL-MODY is a monogenic form of diabetes and pancreatic exocrine dysfunction due to

mutations in the carboxyl-ester lipase gene CEL. The pathogenic mechanism for diabetes

development is unknown. Since CEL is expressed mainly in pancreatic acinar cells, we asked

whether we could find structural pancreatic changes in CEL-MODY subjects during the course

of diabetes development. Furthermore, we hypotesized that the diseased pancreas releases

proteins which are detectable in pancreatic fluid and potentially reflected activation or

inactivation of disease-specific pathways. We therefore investigated non-diabetic and diabetic

CEL mutation carriers by pancreatic imaging studies and secretin-stimulated duodenal juice

sampling. The secretin-stimulated duodenal juice was studied using cytokine assays, triple-stage

mass spectrometry (MS3) and multiplexed mass spectrometry-based measurement of kinase

activities. We identified multiple pancreatic cysts in all the eight diabetic mutation carriers but

not in any of the four non-diabetic mutation carriers or the six healthy controls. Furthermore, we

identified upregulated MAPK target proteins and MAPK-driven cytokines and increased MAPK

activity in the secretin-stimulated duodenal juice. These findings show that subjects with CEL-

MODY develop multiple pancreatic cysts by the time they develop diabetes and that upregulated

MAPK signaling in the pancreatic secretome may reflect the pathophysiological development of

pancreatic cysts and diabetes.

Keywords: MODY; Carboxyl-ester lipase, pancreatic cysts, proteome profiling; MAPK signaling

Page 3 of 61 Diabetes

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INTRODUCTION

Studies of patients with Maturity-onset Diabetes of the Young have provided important insight

into disease mechanisms of pancreatic beta cell failure and diabetes development and also

contributed to novel treatment strategies based on this insight (1; 2). We have previously reported

two families with a human monogenic syndrome of diabetes and pancreatic exocrine dysfunction

(maturity-onset diabetes of the young, type 8, MODY8; CEL-MODY;OMIM #609812) caused

by mutations in the carboxyl-ester lipase (CEL) gene (3). Patients with CEL mutations develop

pancreatic exocrine dysfunction in early childhood (as measured by low fecal elastase levels),

accumulate pancreatic fatty tissue, and develop diabetes and clinical malabsorption in their fourth

decade of life (3; 4). The CEL gene is primarily expressed in pancreatic acinar cells and lactating

mammary tissue, and encodes a digestive enzyme with a role in cholesterol ester digestion (5).

Studies of animal models have not been able to explain the disease mechanism of diabetes

development in CEL-MODY (6; 7), but cellular studies indicate that mutated CEL protein is

misfolded (8). Hence, to gain further insight into the disease mechanism, we further studied

human CEL mutation carriers using radiological methods and secretin-stimulated duodenal juice

to examine the pancreatic proteome (9). We hypotesized that the diseased pancreas releases

proteins which are detectable in pancreatic fluid and potentially reflected activation or

inactivation of disease-specific pathways. To optimize the accuracy of the proteomics studies we

applied triple-stage mass spectrometry, MS3 (10), and complemented this approach with

cytokine assays and MS-based multiplex kinase activity assays (11) to cross-validate our

findings.


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RESEARCH DESIGN AND METHODS

Patients. We have previously reported various clinical characteristics for the CEL mutation

carrying subjects (3). In this study the studied CEL mutation carrying subjects (CEL+) were

denoted with the prefix D if they had manifest diabetes (D1, D2, D3 and D4), with the prefix P if

they had not yet developed diabetes (P1 or P2), or with the prefix C for the patient with

pancreatic ductal adenocarcinoma (C1). The Supplemental Online Appendix contains the

corresponding pedigree information. The non-family controls were denoted with the prefix N

(N1, N2, N3, N4) for controls in the duodenal juice studies. We sampled duodenal juice from the

patients by endoscopy. To enrich for pancreatic factors, we administered intravenous secretin

((Secrelux® Sanofi, Germany. 1 cU/kg, max 70 cU) to the patients and sampled duodenal juice

from 30 to 45 minutes later since it has been shown that there is a peak secretion from pancreas

in this time interval (12; 13). The controls for the secretin-stimulated duodenal juice studies were

recruited from volunteers. The single pancreatic juice specimen from a CEL mutation carrier

with PDAC was collected during a classical Whipple procedure. All studies were approved by

the Norwegian regional committee for research ethics and the IRB at the Joslin Diabetes Center

and performed according to the Helsinki Declaration. We obtained written informed consent

from subjects or their parents. Patients were of North-European descent and recruited from the

Norwegian MODY Registry.

Protocol for pancreatic imaging. MR imaging was performed on a 1.5 T Siemens Avanto

running Syngo MR B17 (Erlagen, Germany) using a 12-channel body coil. For clinical and

anatomical evaluation of the pancreas a standard abdominal protocol was used: Axial TrueFISP

(true Fast Imaging with Steady-state Precision; TR (repetition time) = 2.79 ms, TE (echo time) =

1.17 ms, slice thickness = 4.0 mm, spacing = 0, alfa = 63 degrees, FOV (field of view) = 380 mm

x 380 mm, matrix = 156 x 192, acquisition time = 15 s) and axial fat-saturated T2-weighted


5

imaging (TR = 1000 ms, TE (echo time) = 90 ms, slice thickness = 5 mm, spacing = 0, alfa = 150

degrees, FOV (field of view) = 350 mm x 350 mm, matrix = 256 x 256, acquisition time = 24 s).

For evaluation of the pancreatic and biliary ducts magnetic resonance

cholangiopancreaticography (MRCP) was performed: Coronal T2-weigthed imaging (TR = 2000

ms, TE (echo time) = 691 ms, slice thickness = 1.1 mm, spacing = 0, alfa = 140 degrees, FOV

(field of view) = 280 mm x 280 mm, matrix = 320 x 320, acquisition time = 5.23 min) with

maximum intensity projection (MIP) and multiplanar reformatting (MPR).

Cytokine assay. Forty-two cytokines were measured in secretin-stimulated duodenal juice

samples (sample volume of 25 microliter), using the MILLIPLEX MAP Human

Cytokine/Chemokine - Premixed 42 Plex kit from Millipore (Billerica, MA). This 42-plex kit

was used to measure the concentrations of EGF, Eotaxin, FGF-2, Flt-3 ligand, Fractalkine, G-

CSF, GM-CSF, GRO, IFN-α2, IFN-γ, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17, IL-

1Rα, IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IP-10, MCP-1, MCP-3, MDC

(CCL22), MIP-1α, MIP-1β, PDGF-AA, PDGF-AB/BB, RANTES, TGF-α, TNF-α, TNF-β,

VEGF, sCD40L, sIL-2Rα. All samples were analyzed with a Luminex 200 xMAP system, and

MILLIPLEX Analyst Software (VigeneTech, Carlisle, MA) was used to analyse the results.

Proteolytic degradation assay. A set of 45 synthetic peptide substrates, each at 1 µM, were

incubated with 10 µg pancreatic fluid and reaction buffer containing Tris-Cl (25 mM, pH 7.5),

MgCl2 (7.5 mM), EGTA (0.2 mM), β-glycerophosphate (7.5 mM), Na3VO4 (0.1 mM) and DTT

(0.1 mM). The reaction was incubated at room temperature for 0, 15, 60 min, after which the

reaction was quenched with 100 µL of 1% trifluoroacetic acid (TFA). Twenty five pmol of

internal standard stable-isotope labeled substrate peptides was spiked into the quenched reaction.

The solution was desalted using Sep-Pak C18 50 mg cartridges (Waters) and dried using vacuum


6

centrifugation. The desalted sample was then resuspended in 100 µL of 5% formic acid of which

2 µL was then subjected to analysis by mass spectrometry. Technical duplicates were performed

for each sample at each time point. The peptides from proteolytic degradation assays were

analysed by LC-MS on a high resolution Exactive Orbitrap mass spectrometer (Thermo). Briefly,

a 45 minute gradient was used to separate the peptides from 10-37% solvent B (0.125% formic

acid in acentonitrile) at a flow rate of 300 mL/min. LC-MS data was collected from 350-1500

m/z and the extracted ion chromatograms of the light and heavy peptides were used to quantitate

the absolute kinase activity and the extent of peptide degradation respectively. Data analysis was

performed using Pinpoint (Thermo).

Quantitative proteomics of pancreatic juice using TMT labeling. Pancreatic juice (100 µL)

from 6 subjects was purified using methanol/chloroform precipitation. The purified protein

pellets were each resuspended in 100 µL of 50 mM HEPES, 4M Urea, pH 8.5. The protein

concentration was determined using Bradford protein assay (Pierce). The cysteine residues were

reduced and alkylated with DTT (5 mM) and iodoacetamide (15 mM) respectively prior to

digestion with Lys-C (Wako, Japan) in a 1:100 LysC:protein ratio. Following digestion

overnight, the digest was acidified with formic acid prior to desalting with C18 solid-phase

extraction (Sep-Pak, Waters, Milford, MA). The desalted peptides were then resuspended in 100

µL of 50 mM HEPES, pH 8.5 and each sample was labeled with one of the 6-plex tandem mass

tag (TMT) reagents (Thermo Scientific, Rockford, IL) for one hour at room temperature. The

TMT labeled peptides were purified, fractionated by strong cation exchange chromatography

(SCX) and 14 fractions were analysed by LC-MS/MS on an LTQ-Orbitrap Velos mass

spectrometer. An MS3 method developed recently by the lab was used to overcome the

interference problem in acquisition of TMT data as described (14). Further experimental details

for the proteomics and immunoblotting experiments, kinase activity assay (15) and proteolytic


7

degardation assays are outlined in the Supplemental Online Appendix. Details for the database

searches, filtering and quantification for the proteomics experiments are also outlined in the

Supplemental Online Appendix, but, briefly, two different types of searches (LysC and "no

enzyme") were used when MS/MS spectra were searched against the human ipi database (v. 3.6)

using the Sequent algorithm (v. 5) (16). The data was filtered to a false discovery rate of less than

1% based on the target-decoy database approach (17). Further filtering of the list to include only

'pancreas specific' proteins was based on analyses of nine different mouse tissues (18). To test the

hypothesis that secretin-stimulated duodenal secretory proteins truly reflected pancreatic disease,

an investigator (F.M.) blinded to the sample status, performed unsupervised clustering analysis

based on protein abundance (i.e. algoritmic classification procedure to group samples based on

protein expression similarities and blinded to a knowledge of sample disease status).

Bioinformatics studies using Biobase. We performed a comprehensive investigation of all

proteins in the secretin-stimulated duodenal juice (not only pancreatic specific proteins) to avoid

excluding proteins that are only expressed in the diseased state. We used the Biobase Explain 3.0

tool (19) to examine whether the proteins that were elevated in the secretin-stimulated duodenal

juice from CEL mutation carriers (log2 ratio>1 of protein expression) shared common upstream

key nodes in pathways in physiological and pathophysiological processes, see

http://explain30.biobase-international.com.ezp-prod1.hul.harvard.edu/cgi-

bin/biobase/ExPlain_3.0/bin/start.cgi? for details.

Other statistical analysis. For the analyses of cytokine levels we used two-tailed Student’s t-

tests of independent groups assuming unequal variance. We used linear regression to estimate R2

for the correlation of cyst number and the respective cytokines. We chose a significance level of


8

5% and analysed all data using Stata 11.0 (Stata Statistical Software, Stata Corp., College

Station, Texas, USA).

RESULTS

Subjects with CEL-MODY Develop Multiple Pancreatic Cysts and diabetes in their forties.

We performed pancreatic imaging studies in a CEL-MODY family (Figure 1) and identified

multiple unilocular pancreatic cysts (Figure 2) in all CEL mutation carriers (n=8) who had also

developed diabetes (Table 1). None of the non-diabetic CEL mutation carriers (n=4) or the

healthy controls (n=6) had multiple pancreatic cysts, but two of these subjects (one non-diabetic

CEL mutation carrier and a healthy control) had a solitary cyst. Moreover, the number of cysts

correlated positively with age in the CEL mutation carriers (R2 = 0.46, P = 0.01).

Cytokine analysis of the secretin-stimulated duodenal juice identifies MAPK-driven

cytokines in CEL mutation carriers positively correlated with disease progression. Since

cytokine levels in secretin-stimulated duodenal juice may reflect pancreatic disease (20), we first

assessed a panel of 42 cytokines in the secretin-stimulated duodenal fluid samples of several

family members and controls (Suppl. Table 1). The MAPK-driven CXCR2 receptor targeting

cytokine growth-related oncogene (GRO) was significantly increased in the carriers with a

tendency towards higher levels in subjects who had accumulated more clinical manifestations

(Table 1, and Fig. 3a; P<0.01). We also observed that GRO correlated significantly with the

number of cysts in the subjects (Fig. 3b; R2=0.56, P=0.005). Interestingly, the related MAPK-

driven CXCR1- and 2-targeting cytokine interleukin 8 (IL8), while not showing significantly

increased levels in CEL mutation carriers (Fig. 3c; P=0.15), revealed a significant correlation

with the number of cysts (Fig. 3d; R2=0.72, P<0.001; Suppl. Table 1). Hence, secretin-stimulated


9

duodenal fluid of CEL mutation carriers contains cytokines whose increasing levels may

potentially reflect increasing severity of a progressive pancreatic disease.

Proteomics-based identification of a distinct protein signature in secretin-stimulated

duodenal juice of CEL mutation carriers. Since some protein markers (cytokines) displayed

increasing levels with the progressive accumulation of clinical manifestations, we asked whether

diabetic CEL mutation carriers displayed a unique proteomic signature in secretin-stimulated

duodenal juice. Anticipating some degree of protein degradation in duodenal juice we first

assessed the extent of protein degradation by exploring a multiplexed mass spectrometry based

degradation assay (Fig. 4a) and differences in silver staining for equal quantities of protein

loaded onto gels (Fig. 4b) and found indeed an extensive degradation of secretin-stimulated

duodenal proteins. Subsequently, we applied a multiplexed quantitative proteomics approach

with isobaric labeling (TMT, tandem mass tags) to a training set of secretin-stimulated duodenal

juice from three CEL mutation carriers and three healthy controls. Consistent with the previously

identified protein degradation we found large differences in protein identifications using standard

LysC compared to ‘no-enzyme’ specific searches of proteomics data (Fig 4c). Hence, to identify

both the degraded and non-degraded proteins to account for all the secreted proteins in the

original sample, we further pursued no-enzyme specified database searching, and we identified

757 proteins (1% false discovery rate) in the secretin-stimulated duodenal juice (Suppl. Table 2)

despite the extensive protein degradation. Further filtering of the list to include only proteins

abundantly expressed in mice pancreata (18) resulted in the identification of 115 proteins

(excerpt in Table 2). Intriguingly, an unsupervised clustering analysis clearly separated the

patients and the controls into two separate groups (Fig 5a), supporting the existence of a unique

proteomic signature (Table 2 and Suppl. Table 2) in diabetic CEL mutation carriers. We

identified mostly digestive enzymes (Table 2) among the duodenal proteins showing a relatively


10

higher abundance in control samples (Fig. 5b), including carboxyl-ester lipase (CEL) and

elastase (CELA2A, CELA3B), consistent with our anticipation of a greater abundance of

pancreatic proteases in healthy subjects. Proteins more abundant in CEL-mutation carriers (Table

2) included proteins associated with diabetes (TSPAN8; (21)) and insulin secretion

(GLUD1;(22)). Together these observations provide confidence that secretin-stimulated duodenal

proteins actually reflect pancreatic output and could serve as potential indicators of pancreatic

pathology.

Validation of the training set proteins by immunoblotting and their potential role as

mitogen-activated protein kinase (MAPK) targets. Next, to validate the proteome signature in

CEL mutation carriers, we applied immunoblotting of selected signature proteins in secretin-

stimulated duodenal juice from the training set of three CEL mutation carriers and three controls.

Indeed, band intensities and mass spectrometry abundances correlated well for the proteins

(compare Fig. 5c and Suppl. Table 2), confirming the validity of the mass spectrometry findings.

Furthermore, band intensities were also clearly different between controls and two additional pre-

diabetic CEL mutation carriers (Fig. 5c). Notably, the protein markers LMNA, SKP1 and KRT18

clearly separated CEL mutation carriers from controls, whereas the protein markers PKM2 and

GSN more partially separated these groups. In order to assess whether the proteome signature

proteins in CEL mutation carriers were parts of an activated pathway, we explored the potential

connectivity of the top 30 most abundant proteins in the training set using a curated database

search (Biobase Explain) of reported protein interactions (19). Intriguingly, we found that several

of these validated markers were downstream targets of MAPK (Fig. 5d and Suppl. Table 3),

including several of the proteins also validated by immunoblotting (compare Fig. 5d with Fig.

5c).


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To further assess the strength of connectivity between the most abundant proteins, we used Gene

Network Enrichment Analysis (GNEA; (23; 24)) to test for significant protein interactions based

on yeast two hybrid systems data. We identified the 14-3-3 protein zeta, encoded by YWHAZ,

interacting with Keratin 18 (KRT18) and the subnetwork of interacting proteins around YWHAZ

to be cumulatively significant (Fisher’s p = 0.00028), supporting interactions between 14-3-3

protein zeta and keratin 18 in disease pathogenesis (Suppl. Fig. 1). Both these proteins were also

MAPK targets as defined by the Biobase Explain findings.

Multiplexed kinase studies provide further evidence of altered kinase activity. Since the

above data suggested the involvement of mitogen-activated protein kinase signaling we profiled

multiple kinase activities in both duodenal samples using mass spectrometry ((15); Fig. 5e).

Upon examining a different aliquot of the same secretin-stimulated duodenal juice samples as

that used in the immunoblotting studies, and adopting unsupervised clustering analysis we

observed that the three controls and the pre-diabetic CEL mutation carrier clustered in one group

clearly different from a group with the four diabetic CEL mutation carriers (including a subject

with pancreatic ductal adenocarcinoma). Interestingly, the cell cycle-related kinases, CHK1 and

CHK2, and PKC, and to a lesser extent also MAPK, displayed elevated kinase activity in the

diabetic CEL mutation carriers (Fig. 5f).

DISCUSSION

Our data further support that CEL-MODY is a progressive disease. We have previously reported

the occurence of pancreatic exocrine dysfunction from infancy (3), pancreatic lipomatosis form

early childhood (4) and diabetes developing in the forties (3). Here, we show that CEL mutation

carriers also develop multiple pancreatic cysts as they develop diabetes. Thus, there seems to be a


12

sequence of clinical characteristics over time that may reflect a pathogenic process taking place

in the pancreas and ultimately leading to diabetes. Our data support that this pathogenic process

is associated with upregulated MAPK signaling for the following reasons: First, cytokine

analyses in the pancreatic secretome demonstrated increasing levels of the MAPK-driven (25)

cytokine GRO over the course of the disease. Second, using a sensitive and accurate quantitative

proteomics approach, we identified elevated levels of downstream target proteins of

MAPK/ERK2 signaling in diabetic CEL mutation carriers. Third, the mass spectrometry data

were confirmed by immunoblotting for proteins in the duodenal juice, and immunoblotting also

confirmed our observations in several other CEL mutation carrying subjects. Finally, we

identified elevated kinase activity including that of MAPK in CEL mutation carriers.

Our data also support our hypothesis that secretin-stimulated duodenal secretory proteins truly

reflect pancreatic disease. First, unsupervised clustering analysis of proteomics data correctly

identified the diseased subjects. Second, we also detected increased levels of digestive enzymes

in healthy subjects consistent with our anticipation of a greater abundance of pancreatic proteases

in healthy subjects and consistent with proteomics findings in normal pancreatic juice by ERCP

(26), by enzymatic findings (of elastase) in feces (3) and by enzymatic findings (of amylase,

lipase and chymoptrypsin) in the secretin-stimulated duodenal juice of CEL mutation carriers

(27). Third, the increased levels of digestive enzymes were confirmed by immunoblotting (for

amylase).

We have described a new feature of the CEL-MODY phenotype by demonstrating the presence

of pancreatic cysts in mutation carriers. Such cysts are an established risk factor for pancreatic

cancer (28). Notably, one of the oldest affected members of the family was diagnosed with

pancreatic ductal adenocarcinoma at the age of 78. We therefore conclude that CEL-MODY


13

patients could be at increased risk for developing pancreatic cancer. The cytokine analyses

revealed a linear relationship between the number of cysts and the CXCR1-and CXCR2-targeting

MAPK-driven (25) cytokines (GRO, IL8); a finding consistent with a previous report of elevated

IL8 levels in sera of patients with pancreatic cancer (29). We believe that the low levels of GRO

and IL8 despite multiple cysts in a diabetic CEL mutation carrier (Subject D1) fits with the long

duration of his diabetes and the development of end-stage pancreatic disease with loss of

inflamatory cells as observed in end-stage pancreatitis (30), although there also the possibility of

phenotypic variance within the family. The finding of MAPK-related proteins and cytokines in

duodenal juice of subjects with progressive pancreatic disease is particularly interesting due to

the role of MAPK both in diabetes and cancer (31). In the subject with cancer, the tumor had a

somatic KRAS mutation (G12V, not shown) commonly observed in pancreatic adenocarcinomas

(32). Cancer-associated KRAS mutations generally lead to overactive proteins that stimulate

oncogenic signaling through the MAPK pathway (33).

In conclusion, subjects with CEL-MODY develop multiple pancreatic cysts and diabetes in their

forties. Increased levels of MAPK target proteins may reflect the pathophysiological

development of pancreatic cysts and diabetes in CEL-MODY. These data suggest that the MAPK

pathway should be further explored in subjects with CEL-MODY in order to find drug targets for

the possible prevention of disease development.

ACKNOWLEDGEMENTS

This work was supported by NIH grant RO1 DK 67536 and the Graetz Bridge Funds (to R.N.

Kulkarni), and the Joslin DRC Specialized Assay and Advanced Microscopy Cores (NIH P30


14

DK36836) and NIH grant HG3456 (to S.P.G.) and the Norwegian Research Council (to H.R. and

P.R.N.) and by grants from the Western Norway Regional Health Authority (to H.R., H.I., and

P.R.N.) and the Norwegian Endocrine Society (to H.R.) and the Norwegian Cancer Society (to

A.M.) and the Fulbright Foundation (to H.R.), and the University of Bergen (to P.R.N.), and

Innovest (to P.R.N.).

No potential conflicts of interest relevant to this article were reported.

HR SPG RNK designed the study; HR FEMA ET SB IH JH SMW MV AEO DH GD collected

the data; HR FEMA ET SB IH ML MBR HI PRN AM SPG RNK contributed to data analysis

and interpretation. The manuscript was drafted by HR and RNK with input from all authors. All

authors contributed to the revision and approved the final version of the manuscript.

HR and RNK are the guarantors of this work and, as such, had full access to all data in the study

and take responsibility for the integrity of the data and accuracy of the data analysis.

The authors thank C. Ronald Kahn MD and Chong Wee Liew PhD of Joslin Diabetes Center for

discussions; G. Sankaranarayanan of Joslin Diabetes Center for assistance with cytokine assays;

C. Cahill of Joslin Diabetes Center for assistance with electron microscopy; E. Huttlin of

Harvard Medical School for assistance with data analysis of pancreatic specific proteins.

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C, Aerts J, Ahmad T, Andrews TD, Arbury H, Attwood A, Auton A, Ball SG, Balmforth AJ, Barrett JC, Barroso I, Barton A, Bennett AJ, Bhaskar S, Blaszczyk K, Bowes J, Brand OJ, Braund PS, Bredin F, Breen G, Brown MJ, Bruce IN, Bull J, Burren OS, Burton J, Byrnes J, Caesar S, Clee CM, Coffey AJ, Connell JM, Cooper JD, Dominiczak AF, Downes K, Drummond HE, Dudakia D, Dunham A, Ebbs B, Eccles D, Edkins S, Edwards C, Elliot A, Emery P, Evans DM, Evans G, Eyre S, Farmer A, Ferrier IN, Feuk L, Fitzgerald T, Flynn E, Forbes A, Forty L, Franklyn JA, Freathy RM, Gibbs P, Gilbert P, Gokumen O, Gordon-Smith K, Gray E, Green E, Groves CJ, Grozeva D, Gwilliam R, Hall A, Hammond N, Hardy M, Harrison P, Hassanali N, Hebaishi H, Hines S, Hinks A, Hitman GA, Hocking L, Howard E, Howard P, Howson JM, Hughes D, Hunt S, Isaacs JD, Jain M, Jewell DP, Johnson T, Jolley JD, Jones IR, Jones LA, Kirov G, Langford CF, Lango-Allen H, Lathrop GM, Lee J, Lee KL, Lees C, Lewis K, Lindgren CM, Maisuria-Armer M, Maller J, Mansfield J, Martin P, Massey DC, McArdle WL, McGuffin P, McLay KE, Mentzer A, Mimmack ML, Morgan AE, Morris AP, Mowat C, Myers S, Newman W, Nimmo ER, O'Donovan MC, Onipinla A, Onyiah I, Ovington NR, Owen MJ, Palin K, Parnell K, Pernet D, Perry JR, Phillips A, Pinto D, Prescott NJ, Prokopenko I, Quail MA, Rafelt S, Rayner NW, Redon R, Reid DM, Renwick, Ring SM, Robertson N, Russell E, St Clair D, Sambrook JG, Sanderson JD, Schuilenburg H, Scott CE, Scott R, Seal S, Shaw-Hawkins S, Shields BM, Simmonds MJ, Smyth DJ, Somaskantharajah E, Spanova K, Steer S, Stephens J, Stevens HE, Stone MA, Su Z, Symmons DP, Thompson JR, Thomson W, Travers ME, Turnbull C, Valsesia A, Walker M, Walker NM, Wallace C, Warren-Perry M, Watkins NA, Webster J, Weedon MN, Wilson AG, Woodburn M, Wordsworth BP, Young AH, Zeggini E, Carter NP, Frayling TM, Lee C, McVean G, Munroe PB, Palotie A, Sawcer SJ, Scherer SW, Strachan DP, Tyler-Smith C, Brown MA, Burton PR, Caulfield MJ, Compston A, Farrall M, Gough SC, Hall AS, Hattersley AT, Hill AV, Mathew CG, Pembrey M, Satsangi J, Stratton MR, Worthington J, Deloukas P, Duncanson A, Kwiatkowski DP, McCarthy MI, Ouwehand W, Parkes M, Rahman N, Todd JA, Samani NJ, Donnelly P: Genome-wide association study of CNVs in 16,000 cases of eight common diseases and 3,000 shared controls. Nature 2010;464:713-720 22. Stanley CA, Lieu YK, Hsu BY, Burlina AB, Greenberg CR, Hopwood NJ, Perlman K, Rich BH, Zammarchi E, Poncz M: Hyperinsulinism and hyperammonemia in infants with regulatory mutations of the glutamate dehydrogenase gene. N Engl J Med 1998;338:1352-1357 23. Strnad P, Tao GZ, Zhou Q, Harada M, Toivola DM, Brunt EM, Omary MB: Keratin mutation predisposes to mouse liver fibrosis and unmasks differential effects of the carbon tetrachloride and thioacetamide models. Gastroenterology 2008;134:1169-1179 24. Tao GZ, Li DH, Zhou Q, Toivola DM, Strnad P, Sandesara N, Cheung RC, Hong A, Omary MB: Monitoring of epithelial cell caspase activation via detection of durable keratin fragment formation. J Pathol 2008;215:164-174 25. Ancrile BB, O'Hayer KM, Counter CM: Oncogenic ras-induced expression of cytokines: a new target of anti-cancer therapeutics. Molecular interventions 2008;8:22-27 26. Zhou L, Lu Z, Yang A, Deng R, Mai C, Sang X, Faber KN, Lu X: Comparative proteomic analysis of human pancreatic juice: methodological study. Proteomics 2007;7:1345-1355 27. Tjora E, Wathle G, Engjom T, Erchinger F, Molven A, Aksnes L, Haldorsen IS, Dimcevski G, Njolstad PR, Raeder H: Severe Pancreatic Dysfunction But Compensated Nutritional Status in Monogenic Pancreatic Disease Caused by Carboxyl-Ester Lipase Mutations. Pancreas 2013; 28. Tanaka S, Nakao M, Ioka T, Takakura R, Takano Y, Tsukuma H, Uehara H, Suzuki R, Fukuda J: Slight dilatation of the main pancreatic duct and presence of pancreatic cysts as predictive signs of pancreatic cancer: a prospective study. Radiology 2010;254:965-972 29. Wigmore SJ, Fearon KC, Sangster K, Maingay JP, Garden OJ, Ross JA: Cytokine regulation of constitutive production of interleukin-8 and -6 by human pancreatic cancer cell lines and serum cytokine concentrations in patients with pancreatic cancer. Int J Oncol 2002;21:881-886


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18

TABLES

Status Sex Age

(years)

Fecal

elastase

(ug/g)

Diabetes?

Diabetes

duration

(years)

Number

of cysts

Largest

cyst

diameter

(mm)

Pancreatic

duct>2.5

mm?

GRO

(pg/mL)

IL8

(pg/mL)

Control F 32 426 no 0 0 0 no 2.5 0.3 Control (N2) F 38 561 no 0 0 0 no 7.3 0.0

Control M 40 584 no 0 0 0 no 5.9 1.5 Control F 50 723 no 0 0 0 no 0.0 0.2 Control F 53 486 no 0 1 2 no 2.5 0.1 Control F 64 525 no 0 0 0 no 9.7 2.1

Control (N1) M 63 363 no 0 NA NA NA 4.3 2.6 Control (N3) M 25 299 no 0 NA NA NA 11.3 4.4 CEL carrier M 9 448 no 0 0 0 no 2.5 2.0 CEL carrier M 14 50 no 0 0 0 no 11.1 4.5 CEL carrier F 17 71 no 0 1 6 no 5.1 5.3 CEL carrier

(P2) M 37 10 no 0 0 0

no NA NA CEL carrier

(P1) M 40 12 no 0 0 0

no 7.3 7.7 CEL carrier

(D2) F 43 0 yes 13.1 12 7

no 116.7 21.1 CEL carrier F 45 51 yes 6.2 9 4 no 45.1 16.8 CEL carrier M 56 0 yes 11.8 6 4 no 97.3 4.3 CEL carrier M 57 0 yes 17.5 4 11 yes 15.2 3.1 CEL carrier

(D4) F 57 5 yes 19.1 60 11


(D1) M 65 0 yes 44.7 20 13


(D3) M 69 3 yes 13.6 30 9


(C1) F 78 0 yes 37.5

60 13 no 161.0 167.7

Table 1: Clinical characteristics of CEL mutation carriers and controls. NA, not available.

The alphanumeric codes i Status are explained in Methods.


19

Gene ID Log 2 ratio Log 2 total Gene ID Log 2 ratio Log 2 total

1 CLEC3B -16.0 23.2 87 MUC1 2.4 20.2 2 KRT18 -15.2 22.4 88 CUZD1 2.5 25.8 3 NUCB2 -12.5 19.7 89 CLPS 2.6 22.5 4 DCI -11.5 19.6 90 PNLIPRP2 2.7 27.3 5 MYL1 -11.5 18.7 91 KLK1 2.7 24.1 6 RPLP2 -11.1 23.2 92 RNASE1 3.5 25.7 7 SKP1 -10.8 18.1 93 CPA2 3.6 27.7 8 RPLP0 -10.7 21.7 94 S100A7 3.9 22.4 9 RPLP1 -9.7 21.4 95 CEL 3.9 28.5

10 SFN -9.5 19.1 96 CTRB1 3.9 27.8 11 EPB41L3 -9.4 22.1 97 CTRL 3.9 23.9 12 CHGA -8.9 16.1 98 PRSS1 4.0 28.1 13 PKM2 -7.5 22.3 99 CPA1 4.1 28.9 14 IDH1 -7.1 22.5 100 CELA3B 4.3 25.6 15 MYH14 -6.2 22.3 101 PNLIPRP1 4.3 26.3 16 PURA -6.2 20.7 102 IPI00219910 4.6 23.5 17 LMNA -5.5 21.3 103 PNLIP 4.8 30.0 18 KRT8 -5.1 24.9 104 PRSS2 4.8 26.6 19 EEF1A1 -5.1 21.0 105 CELA3A 4.9 27.8 20 GLUD1 -4.9 22.5 106 CTRC 5.0 26.8 21 PCDH24 -4.7 21.8 107 GP2 5.2 25.8 22 PPA1 -4.1 22.1 108 CPB1 5.3 27.9 23 NPC2 -4.0 20.2 109 AMY1C 5.3 26.8 24 ALDOB -3.8 25.5 110 AMY2B 5.4 29.6 25 TSPAN8 -3.7 20.2 111 SERPINI2 6.7 22.8 26 CLIC1 -3.7 23.0 112 PLA2G1B 6.9 25.8 27 PGD -3.7 19.9 113 CD302 8.2 16.7 28 CST3 -3.3 23.5 114 HPN 8.4 18.4 29 GSN -3.3 24.9 115 CTRB2 11.3 18.5

Table 2: The most extreme differentially expressed proteins in secretin-stimulated duodenal

juice. Log2 ratio, Log2 of the ratio of mass spectrometry abundance for the sum of the three

healthy controls divided by the sum of the three diabetic mutation carriers. Log 2 total, Log 2 of

the total mass spectrometry abundance for a protein as the sum of the abundances of three

healthy controls and the three diabetic mutation carriers.


20

FIGURE LEGENDS

Fig. 1: Overview of the multimodal systems biology approach.

A systems biology approach using secretin-stimulated duodenal juice from subjects in a CEL-

MODY family to discover early markers in pancreatic disease development by proteomics

methods.

Fig. 2: The Magnetic Resonance Imaging of Pancreatic cysts in CEL mutation carriers.

MRCP (a-c) and axial fat-saturated T2-weighted imaging at the level of the pancreatic body (d-f)

in two CEL-mutation carriers (a, d and b, e) and a normal control (c, f). Both mutation carriers

had multiple cystic lesions within the pancreatic body and tail (white thin arrows; a-b, d-e) as

well as in the pancreatic head (arrowheads; a-b). The pancreatic ducts (filled black arrows) and

common bile ducts (filled white arrows) have normal caliper (a-b, d-e) similar to that in the

control (c, f). GB, gallbladder. The alphanumeric codes refer to the subject designation explained

in Research Design and Methods.

Fig. 3: The identification of a patient-specific cytokine signature in pancreatic juice and a

mass spectrometry degradation assay.

Cytokine levels for GRO compared to mutation status (a) and correlated with cyst number (b).

Cytokine levels for IL-8 compared to mutation status (c) and correlated with cyst number (d).

Further, IL-1b was increased in the carriers (P=0.005) and also significantly correlated with the

number of cysts (R2=0.70, P<0.001). None of the remaining cytokines correlated significantly


21

with the number of cysts although significantly elevated in the carriers (FGF-2 (P=0.04), Flt3-

ligand (P=0.03), GM-CSF (P=0.01), IL-1ra (P=0.02), IL-12 (P=0.01), MCP-1(P=0.04), s-CD-

40L (P=0.03); Suppl. Table 1).

Fig. 4: Reduced substrate degradation in CEL-MODY patients.

Greater substrate degradation was observed for control patients compared to CEL-MODY

patients. A set of 45 substrate peptides are incubated with pancreatic juice for 45 minutes and the

amount of degradation is measured by comparison of the substrate with internal standard heavy

peptides at 0, 15, 60 minute time points (a). The differences in protein identifications using silver

staining of secretin-stimulated duodenal juice for equal quantities of protein loaded onto gels (b).

Consistent with this reduced substrate degradation in the secretin-stimulated duodenal juice of

CEL-MODY patients we also identified differences in protein identifications using LysC and

‘no-enzyme’ specific searches of proteomics data (c). Using ‘no-enzyme’ specific searches we

identified 757 proteins in the secretin-stimulated duodenal juice.

Fig. 5: The identification of a patient-specific protein signature in pancreatic juice

Protein quantitation was performed using isobaric labeling with tandem mass tags (TMT) and

LC-MS/MS analysis on an LTQ-Orbitrap Velos mass spectrometer. An unsupervised clustering

analysis identified clearly different expression patterns in CEL mutation carriers compared to

controls with analyses performed by an investigator (FM) blinded to the samples (a). Log2-plot

of secretin-stimulated duodenal proteins protein abundances and ratios of levels in the three CEL

mutation carriers compared to the three controls (b). Log2 ratio, Log2 of the ratio of mass

spectrometry abundance for the sum of the three controls divided by the sum of the three diabetic

mutation carriers. Log 2 total, Log 2 of the total mass spectrometry abundance for a protein as

the sum of the abundances of three controls and the three diabetic mutation carriers. Note the


22

lack of skewed distribution of pancreatic proteins in the log2-plot indicating that a majority of the

proteins had been detected despite degradation. Using ‘no-enzyme’ specific searches we

identified 757 proteins in the secretin-stimulated duodenal juice, of which 115 proteins were

pancreatic specific based on (18). Immunoblot validation examples of proteins identified with

stipled lines indicate gel crops (c). In addition to the three diabetic CEL mutation carriers (D1,

D2, D3) and three non-family controls (N1,N2,N3) we also added secretin-stimulated duodenal

juice samples from two pre-diabetic CEL mutation carriers (subjects P1 and P2) and from the

CEL mutation carrier with the KRAS-mutated pancreatic ductal adenocarcinoma (Subject C1).

Visualization of the MAPK-targeted proteins identified by Biobase, ExPlain 3.0 tool, and

alphanumeric codes refering to pubmed IDs in Suppl. Table 3 (d). Heatmap demonstrating

differential clustering of kinase activities in the secretin-stimulated duodenal juice and pancreatic

tissue of CEL mutation carriers and controls (e). Vertical bar graphs showing significant

differences in kinase activities for PKC, CH1 and CH2 in secretin-stimulated duodenal juice and

for PKC in pancreatic tissue. The multiplexed kinase activity assay allows the absolute

quantitation of kinase activities by measuring the amount of phosphorylation of 60 peptide

substrates with known motifs for many kinases. (f) Vertical bar graphs showing differences in

kinase activities for PKC, CH1, CH2 and MAPK in secretin-stimulated duodenal juice. PDAC,

pancreatic ductal adenocarcinoma.


42x9mm (300 x 300 DPI)


110x60mm (300 x 300 DPI)


146x116mm (300 x 300 DPI)


152x141mm (300 x 300 DPI)


Fig.5

246x235mm (300 x 300 DPI)


1

Supplemental Online Appendix Raeder et al; Carboxyl-ester lipase Maturity-Onset Diabetes of the Young is Associated with Development of Pancreatic Cysts and Upregulated MAPK Signaling in Secretin-stimulated duodenal fluid

Supplementary Material Patients

The subject designation corresponds to the following pedigree status in a published

pedigree (1): D1 = III-9, D2 = IV-11, D3 = III-5, P1 = IV-8, P2 = IV-12.

Sample preparation for tissue samples for kinase activity and proteolytic

degradation assays

Pancreatic tissue was homogenized using a Dounce homogenizer and ice-cold lysis

buffer: 10 mM K2HPO4 pH 7.5, 1 mM EDTA, 10 mM MgCl2, 50 mM β-

glycerophosphate, 5 mM EGTA, 0.5% Nonidet P-40, 0.1% Brij 35, 0.1% deoxycholic

acid, 1mM sodium orthovanadate, 1mM phenylmethyl-sulfonyl fluoride and protease

inhibitors (Complete protease inhibitor tablet, Roche). Following homogenization, the

lysate was centrifuged at 10,000 rpm for 10 min at -4 °C and the supernatant was snap

frozen with liquid nitrogen and stored at -80 °C. Kinase activities were determined on

lysate that had only been frozen once following harvesting since freeze-thaw cycles

decrease kinase activities. Protein concentration was determined by the modified

Bradford assay (Pierce).

Proteomic analysis of pancreatic fluid

The purified peptides following digestion were labeled with TMT. Each tube of 0.8

mg for each TMT label was dissolved in 40 µL of anhydrous acetonitrile, of which 5


2

µL of each was used to label peptides from 40 µg of pancreatic fluid protein. The

labeling reaction was quenched by the addition of 10 µl of 5% hydroxylamine and the

individual samples combined and desalted with C18 solid-phase extraction (Sep-Pak,

Waters, Milford, MA).

The combined sample was fractionated by strong cation exchange chromatography

(SCX), similar to that described previously (2). Briefly, a 2.1 x 200 mm

polySULFOETHYL A HPLC column (particle size, 5 µm; pore size, 200 Å; PolyLC,

Columbia, MD) was used to separate the sample that was resuspended in 300 µL SCX

buffer A (7 mM KH2PO4, 30% ACN, pH 2.6). A two-buffer (SCX A and B) gradient

from 0 to 50% SCX buffer B (7 mM KH2PO4, 350 mM KCl, pH 2.6, 30% ACN) in

47 min was used at a flow rate of 0.2 ml/min, followed by 50 to 100% SCX buffer A

to buffer B in 4.5 min using an Agilent 1100 quaternary pump with a degasser and a

photodiode array detector (PDA) (Thermo Scientific, San Jose, CA). Fractions were

collected every 90 s into a 96-well plate, and dried with a speed-vac. The dried

peptide fractions were then resuspended with 1% FA, and combined into 14 fractions

based on the intensity from the SCX chromatographic UV trace and then desalted by

C18 SPE, and dried using a speed-vac.

Dried peptides were resuspended in 15 µL 4% formic acid, 5% ACN, and 2 µL was

loaded onto a microcapillary column (100 µm I.D, 20 cm, packed with Magic C18AQ

resin: 5 µ m, 100 Å, Michrom Bioresources, Auburn, CA) with Famos autosampler

(LC Packings, Sunnyvale, CA) and an Agilent 1100 binary HPLC pump (Agilent

Technologies, Santa Clara, CA). Reverse phase chromatography was used for peptide

separation with a 150 minute gradient of 9 to 32% ACN in 0.125% FA at a flow rate

of around 300 nL/min. An LTQ Orbitrap Velos (Thermo Fischer Scientific) was


3

used, with XCalibur 2.0.7 acquisition software. Data acquisition was performed in

data-dependent mode with a survey scan in the range of 300-1500 m/z with a 30,000

resolution. The top 10 most intense ions were selected for fragmentation in the LTQ

with a precursor isolation width window of 2 m/z. The AGC target was set at 3x106

for the survey scan and ions were selected for MS2 and their intensity was greater

than 500 accounts. Ions with unassigned or 1+ charge states were excluded. Selected

ions were excluded from further analysis for 30 s. Maximum ion accumulation times

were 1000 ms and 250 ms for survey and MS/MS scans respectively. An MS3 method

developed by the lab was used to overcome the problem of interfering peaks in the

analysis of TMT data. The instrument parameters used were the same as those

described (3). Briefly, after each MS2 analysis, the most intense fragment ion in an

m/z range between 110-160% of the precursor m/z was selected for HCD-MS3. The

fragment ion isolation width was set to 4 m/z, the MS3 AGC was 20 x 104 and the

maximum ion time set to 250 ms. The normalized collision energy was set to 35%

and 60% at an activation time of 20 ms and 50 ms for MS2 and MS3 scans,

respectively.

Database searches and data filtering and quantitation

Following mass spectrometric acquisition, the RAW files were converted into the

mzXML format and individual precursors selected for MS2 fragmentation were

checked for erroneous monoisotopic m/z and the mass measurements refined (4). The

MS/MS spectra were searched against the human ipi database (version 3.6) using the

Sequest algorithm (version 28) (5). A decoy database search approach was employed,

where each protein sequence was listed in both the forward and reversed direction, to

enable the estimation of the peptide and protein identification false discovery rates


4

(FDR). The search parameters were as follows: 50 ppm precursor ion tolerance; 1.0

Da product mass tolerance; variable modifications:oxidation of methionine

(+15.9949); fixed modifications: carbamidomethylation of cysteine (+57.0214), Six-

plex TMT tags on lysine residues and peptide N termini (+ 229.162932 Da). Two

different types of searches were performed. Firstly, LysC specificity was required and

secondly, 'no enzyme specificity' was required. On account of the large numbers of

proteolytic enzymes known to be present in pancreatic fluid, it is likely that during the

clinical sampling of pancreatic fluid degradation by enzymes had occurred in addition

to the desired Lys-C digestion, hence the reason to search with 'no enzyme' specified'.

The data was filtered to a false discovery rate of less than 1% based on the target-

decoy database approach (6). Filtering using linear discriminant analysis was

performed to generate a classifier to distinguish between correct and incorrect

identifications based on the following parameters: XCorr, ∆Cn, peptide ion mass

accuracy, charge state and peptide length, as described (4).

Further filtering of the list to include only 'pancreas specific' proteins was based on

analyses of nine different mouse tissues (4)

(https://gygi.med.harvard.edu/phosphomouse/index.php).

To test the hypothesis that secretin-stimulated duodenal secretory proteins truly

reflected pancreatic disease, an investigator (F.M.) blinded to the samples, performed

unsupervised clustering analysis based on protein abundance (i.e. algoritmic

classification procedure to group samples based on protein expression similarities and

blinded to a knowledge of sample disease status).

Peptide quantification using TMT reporter ion intensity was performed using in-house

software, as described previously (3). Briefly, a 0.06 m/z window around the

theoretical m/z value of each reporter ion (126, 127, 128, 129, 130 and 131) was


5

scanned for ions, and the intensity of the signal nearest to the theoretical m/z value

was recorded. The intensities of the reporter ion intensities were denormalised by

multiplying them with the ion accumulation time for each MS3 spectrum and.

Adjustment based on the overlap of isotopic envelopes for the reporter ions was

performed.

Immunoblotting. Secretions from the pancreas (15 microgram applied to each well;

concentration estimated by BCA assay; Pierce) was applied directly to and separated

by SDS-PAGE, electrophoretically transferred to a nitrocellulose membrane and

stained with anti-keratin 18 antibody (4548S ;1:2000; Cell Signaling Technology),

anti-SKP1 antibody (2156S; 1:500; Cell Signaling Technology), anti-PKM2 antibody

(4053S; 1:1000; Cell Signaling Technology), anti-LMNA (4777S;1:1000; Cell

Signaling Technology), anti-GSN (AB2969; 1:2000; Millipore) or anti-amylase

(3796S;1:500; Cell Signaling Technology). Staining was developed using a

chemoilluminescence method (Pierce ECL Western blotting substrate; Pierce).

Kinase activity assay

The procedures were similar to that described (7) but with a few changes to the

method as described below. A set of 60 synthetic peptide substrates, each at 5 µM,

were incubated with 10 µg lysate and kinase reaction buffer containing Tris-Cl (25

mM, pH 7.5), ATP (5 mM), MgCl2 (7.5 mM), EGTA (0.2 mM), β-glycerophosphate

(7.5 mM), Na3VO4 (0.1 mM) and DTT (0.1 mM). After incubation for 45 minutes at

room temperature, the reaction was quenched with 100 µL of 1% trifluoroacetic acid

(TFA). A known amount (5 pmol) of internal standard phosphorylated stable-isotope

labeled peptides were spiked into the quenched reaction. The solution was desalted


6

using Sep-Pak C18 50 mg cartridges (Waters) and dried using vacuum centrifugation.

Different to the previous method (7), enrichment for phosphoproteins using

immobilized metal affinity chromatrography (IMAC) was not performed. The

desalted sample were resuspended in 100 µL of 5% formic acid of which 2 µL was

then subjected to analysis by reverse phase liquid chromatography- mass

spectrometry. Technical duplicates were performed for each lysate.

Kinase abbreviation

Category Kinase

AA PKA

CC PKC

DD Acidic

MM MAPK

PP Proline directed

RR RSK

KK AKT

XX pY_group_1

YY pY_group_2

ZZ pY_group_3

QQ DNA damage

GG GSK3

CH CHEK

OO Other S/T kinase

WW Y other


7

BB Basic

AM AMPK

Proteolytic degradation assay

A set of 45 synthetic peptide substrates, each at 1 µM, were incubated with 10 µg

pancreatic fluid and reaction buffer containing Tris-Cl (25 mM, pH 7.5), MgCl2 (7.5

mM), EGTA (0.2 mM), β-glycerophosphate (7.5 mM), Na3VO4 (0.1 mM) and DTT

(0.1 mM). The reaction was incubated at room temperature for 0, 15, 60 min, after

which the reaction was quenched with 100 µL of 1% trifluoroacetic acid (TFA).

Twenty five pmol of internal standard stable-isotope labeled substrate peptides was

spiked into the quenched reaction. The solution was desalted using Sep-Pak C18 50

mg cartridges (Waters) and dried using vacuum centrifugation. The desalted sample

was then resuspended in 100 µL of 5% formic acid of which 2 µL was then subjected

to analysis by mass spectrometry. Technical duplicates were performed for each

sample at each time point.

The peptides from the kinase activity and proteolytic degradation assays were

analysed by LC-MS on a high resolution Exactive Orbitrap mass spectrometer

(Thermo). The experimental details are outlined in the Suppl. information. Briefly, a

45 minute gradient was used to separate the peptides from 10-37% solvent B (0.125%

formic acid in acentonitrile) at a flow rate of 300 mL/min. LC-MS data was collected

from 350-1500 m/z and the extracted ion chromatograms of the light and heavy

peptides were used to quantitate the absolute kinase activity and the extent of peptide

degradation respectively. Data analysis was performed using Pinpoint (Thermo).


8

Mass spectrometry and data analysis for kinase and proteolytic degradation

assays

The peptides from the kinase activity and proteolytic degradation assay, dissolved in

5% formic acid, were analysed by LC-MS on a high resolution Exactive Orbitrap

mass spectrometer (Thermo) equipped with a Nanospray II electrospray ionisation

source (Thermo) coupled to a Thermo binary pump and a Famos autosampler (LC

Packings). Peptides were separated on a hand-pulled fused silica microcapillary with

an internal diameter of 175 µm, 15 cm long, packed with a C18 reversed-phase resin

(Magic C18AQ, particle size 5 µm, pore size 200 Å, Michrom Bioresources, Auburn,

CA). The needle tip had an internal diameter of approximately 5 µm. Once loaded,

the peptides were separated across a 45 minute linear gradient of 10-37% solvent B

(0.125 % FA in ACN) at a flow rate of 300 nl/min provided across a flow splitter by

the HPLC pumps. Buffer A comprised 0.125% FA and 3% ACN in water. Data was

collected from 350-1500 m/z full MS with a resolution setting of 60,000.

Analysis was performed using the commercial software package Pinpoint

(Thermo).The amount of substrate phosphorylation was determined by comparing the

area of the light phosphorylated substrate peptide with that of the heavy

phosphorylated internal standard peptide. The phosphorylated substrate peptides are

chemically identical to the heavy internal standard and therefore should have the same

chromatographic characteristics and ionisation efficiency. In the case of the

proteolytic degradation assay, the amount of peptide degradation was determined by

comparing the area of the light substrate (degraded during the reaction) with that of


9

the heavy substrate internal standard peptide that was added following quenching of

the reaction. Extracted ion chromatograms were generated from the MS scan with ±

10 ppm mass accuracy around the monoisotopic peak of the most abundant charge

state. The extracted ion chromatograms were integrated using Pinpoint. Whilst

Pinpoint generates the area of the extracted ion chromatograms semi-automatically,

each chromatogram required manual inspection to ensure that the correct boundaries

had been selected. Where this was not the case, the peak boundaries were manually

altered. It was required that heavy and light peptides co-eluted perfectly for accurate

quantitation. Measurements were excluded where the peak height was less than 10000

or where the peak area was less than 1% of the internal standard (50 fmol). Since a

known amount of internal standard was added to the reaction (5 pmol for the kinase

assay, 25 pmol for the proteolytic degradation assay) prior to purification, the

absolute amount of product formed could be determined. The product formed/kinase

activity is presented with units of fmol/µg/min. The ratio of the light to heavy peptide

area was divided by the amount of lysate (10 µg) and the time of the reaction (45 min

for the kinase activity assay). For the proteolytic digestion assay, the degradation is

normalized to the light peptide/heavy peptide ratio for the zero time point.

References

1. Ræder H, Johansson S, Holm PI, Haldorsen IS, Mas E, Sbarra V, Nermoen I, Eide SA, Grevle L, Bjørkhaug L, Sagen JV, Aksnes L, Søvik O, Lombardo D, Molven A, Njølstad PR: Mutations in the CEL VNTR cause a syndrome of diabetes and pancreatic exocrine dysfunction. Nat Genet 2006;38:54-62 2. Villen J, Gygi SP: The SCX/IMAC enrichment approach for global phosphorylation analysis by mass spectrometry. Nature protocols 2008;3:1630-1638 3. Ting L, Rad R, Gygi SP, Haas W: MS3 eliminates ratio distortion in isobaric multiplexed quantitative proteomics. Nature methods 2011;


10

4. Huttlin EL, Jedrychowski MP, Elias JE, Goswami T, Rad R, Beausoleil SA, Villen J, Haas W, Sowa ME, Gygi SP: A tissue-specific atlas of mouse protein phosphorylation and expression. Cell 2010;143:1174-1189 5. Eng JK, McCormack AL, Yates JR, 3rd: An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database. J Am Soc Mass Spectrom. 1994;5:976-989 6. Elias JE, Gygi SP: Target-decoy search strategy for increased confidence in large-scale protein identifications by mass spectrometry. Nature methods 2007;4:207-214 7. Kubota K, Anjum R, Yu Y, Kunz RC, Andersen JN, Kraus M, Keilhack H, Nagashima K, Krauss S, Paweletz C, Hendrickson RC, Feldman AS, Wu CL, Rush J, Villen J, Gygi SP: Sensitive multiplexed analysis of kinase activities and activity-based kinase identification. Nat Biotechnol 2009;27:933-940

Supplementary Figure Legends and Supplementary Table Legends

Suppl. Fig. 1: Protein-protein interaction modeling of upregulated pancreatic juice

proteins in CEL mutation carriers. Using Cytoscape, the figure shows major protein-

protein interactions among the proteins identified in pancreatic juice based on Gene

Network Enrichment Analysis. Note the direct interaction between YWHAZ and

KRT18, two proteins that were identified in the pancreatic juice protein analyses.

More generally, the immediate set of protein-protein interactors to the YWHAZ-

encoded protein, comprising the proteins encoded by YWHAE, ATP5A1, EPB41L3,

PPP1CC, ENO1, and KRT18 was together highly significant (Fisher’s p-value =

0.00028).

Suppl. Table 1: Clinical, imaging and cytokine characteristics for CEL mutation

carriers and controls.

Suppl. table 2: Protein expression values for proteins identified in the pancreatic

proteome by proteomics analysis. Log2 ratio, Log2 of the ratio of mass spectrometry


11

abundance for the sum of the three healthy controls divided by the sum of the three

diabetic mutation carriers. Log 2 total, Log 2 of the total mass spectrometry

abundance for a protein as the sum of the abundances of three healthy controls and the

three diabetic mutation carriers.

Suppl. table 3: PubMed IDs documenting the literature support of pathways

involved in the MAPK-targeted proteins based on the output from the Biobase

Explain tool.

Supplementary Figure and Tables

Suppl. Fig. 1


12

Suppl Table 1:

Status Control Control

(N2)

Control Control Control Control Control

(N1)

Control

(N3)

CEL

carrier

CEL

carrier

Sex F F M F F F M M M M

Age (yrs) 32 38 40 50 53 64 63 25 9 14

EGF 442.88 110.76 20.13 39.72 110.37 63.48 143.63 128.28 101.22 32.19

EOTAXIN 0 1.55 1.07 0 0 1.58 1.79 7.26 1.18 0.53

FGF-2 0 2.5 0 0 0 8.66 2.50 16.71 2.5 0

Flt-3

ligand

1.03 0 0 0 0 3.85 2.49 7.86 1.77 3.18

fractalkine 0 0 0 0 0 0 <1.28 <1.28 0 0

G-CSF 1.11 0 0 0 0 0 <0.42 0.59 0 1.11

GM-CSF 0 0.72 0.96 0 0 0.82 0.74 1.49 2.17 0.74

GRO 2.46 7.32 5.91 0 2.46 9.73 4.32 11.29 2.46 11.05

IFN-a2 0 0 0 0 0 0 <1.05 <1.05 0 0

IFN-G 0 0 0 0 0 1.68 <1.38 2.92 0 0

IL-1a 2.08 0.56 20.53 2.32 0.68 16.15 510.68 79.36 35.79 5.48

IL-1b 0 0 0 0 0 0.76 8.71 1.97 1.14 3.67

IL-1ra 0.79 1.77 7.37 1.91 0 3.51 67.50 10.07 6.38 13.15

IL-2 0 0 0.45 0 0 0.45 <0.45 2.06 0 0

IL-3 4 16.28 4.94 11.56 0 8.52 6.26 12.45 0 0

IL-4 0 0 0 0 0 0 <0.84 <0.84 0 0

IL-5 0 0 0 0 0 0 0.18 <0.16 0 0.2

IL-6 0 0 0 0 0 0.52 1.20 5.19 0 4.75

IL-7 0 0 0 0 0 0 <0.62 <0.62 0 3.56

IL-8 0.31 0 1.52 0.22 0.14 2.13 2.64 4.40 1.95 4.53

IL-9 0 0 0 0 0 0 <1.12 <1.12 0 0

IL-10 0 0 0 0.8 0 0.39 6.40 2.25 0 9.45

IL-

12(P40)

0 0.9 0 1.04 0 3.21 5.02 10.94 6.13 7.25

IL-

12(P70)

0 0 1.28 0 0 0 <1.23 3.93 0 0

IL-13 0 0 0 0 0 0 2.81 2.02 0 3.47

IL-15 0 0 1.99 0 1.85 2.84 2.70 5.41 0 1.65

IL-17A 0 0 0 0 0 0 <0.36 0.90 0 0

IP-10 0 0 0 0 0 0 <0.41 7.08 0 0

MCP-1 16.4 1.07 1.07 0 0 1.07 <0.38 1.40 104.43 23.22

MCP-3 3.64 3.64 4.49 0 0 8.15 4.49 12.96 0 7.49

MDC 3.25 2.32 7.74 0 0 10.24 <1.98 14.83 0 13.36

MIP-1a 0 1.73 7.1 0 0 15.78 14.93 22.33 0 18.64

MIP-1b 0 0 0 0 0 0 <0.41 4.40 0 0


13

PDGF-AA 75.46 69.6 64.7 73.02 135.13 118.56 149.74 73.18 58.35 109.98

PDGF-

AB/BB

0 0 0 0 0 0 19.27 10.71 0 0

RANTES 17.26 0 0 0 0 0 <1.51 <1.51 1.77 7.38

sCD-40L 1.56 0.99 0.99 0.51 0.99 1.88 0.51 1.41 1.26 2.21

sIL-2Ra 0 1.9 0 0 0 2.12 <0.7 3.44 0 0

TGF-a 1.4 0 0 0.63 0.7 0 <0.82 1.28 0 0

TNF-a 0 0 0 0 0 0 <0.42 <0.42 0 0

TNF-b 0 0 0 0 0 0 <0.6 <0.6 0 1.41

VEGF 5.55 0 5.55 0 0 12.51 <0.88 16.84 2.02 12.51

Status CEL

carrier

(P2)

CEL

carrier

(P1)

CEL

carrier

(D2)

CEL

carrier

CEL

carrier

CEL

carrier

CEL

carrier

(D4)

CEL

carrier

(D1)

CEL

carrier

(D3)

CEL

carrier

(C1)

Sex M M F F M M F M M F

Age (yrs) 37 40 43 45 56 57 57 65 69 78

EGF NA 44.74 80.68 23.95 5.5 13.84 125.19 1.76 6.85 19.4

EOTAXIN NA 1.07 0.92 0.1 68.8 0.41 0.57 3.24 40.57 0.71

FGF-2 NA 3.9 19.62 0 14.3 7.57 10.2 3.9 12.57 13.45

Flt-3

ligand

NA 0 6.26 0 6.81 3.18 1.77 1.03 5.69 1.77

fractalkine NA 0 0 0 26.05 0 0 0 487 0

G-CSF NA 0 0.59 0 26.4 3.86 0 0 0 1.95

GM-CSF NA 0.35 1.73 0.19 1.33 0.77 0.77 1.54 2.98 1.58

GRO NA 7.32 116.7 45.11 97.31 15.2 109.7 2.46 88.81 161.02

IFN-a2 NA 0 7.32 0 2.1 0 0 0 0 3.9

IFN-G NA 0 0 0 0 0 0 0 1.45 6.45

IL-1a NA 33.67 273.25 109.59 14.57 37.1 594.34 10.72 6.5 2.16

IL-1b NA 2.39 2.82 1.5 0.67 0 1.24 0 0.76 0

IL-1ra NA 16.26 22.85 186.32 69.2 150.09 41.18 7.71 43.41 15.74

IL-2 NA 0.5 0.54 0 0.74 0.74 0.54 0.45 1.22 0.74

IL-3 NA 0 0 0 0 0 0 5.64 0 0

IL-4 NA 0 2.1 0 3.3 0 0 0 0 0

IL-5 NA 0 0.28 0 0 0 0 0 0 0

IL-6 NA 0 5.19 0 0 0 0 0 0 15.46

IL-7 NA 0 1.36 0 6.23 0 0 0 0 0

IL-8 NA 7.67 21.1 16.76 4.28 3.08 373.59 1.16 4.4 167.74

IL-9 NA 0 0 0 0 0 0 0 0 0

IL-10 NA 0.2 1.53 0.86 0.5 0.39 0 0 0.2 0

IL-

12(P40)

NA 2.16 18.45 0 4.29 1.83 1.5 7.8 5.02 0

IL-

12(P70)

NA 0 1.44 0 0 0 0 0 0 0


14

IL-13 NA 0 2.33 0 11.32 0 0 0 0 0

IL-15 NA 1.85 5.96 0 3.14 3.28 0 4.49 3.21 0

IL-17A NA 0 0 0 0 0 0 0.39 1.47 0

IP-10 NA 0 50.3 0 333.95 23.96 13.9 3.89 165.19 1.12

MCP-1 NA 0 119.07 10.28 225.49 48.74 34.65 5.89 441.82 61.38

MCP-3 NA 6.79 14.62 3.64 6.79 7.49 5.3 3.64 11.14 6.43

MDC NA 4.18 24.01 5.09 16.26 28.57 6.87 3.25 21.56 3.25

MIP-1a NA 4.31 22.97 0 18.39 0 0 10.42 26.41 3.6

MIP-1b NA 0 14.68 0 19.84 0 0 0 39.28 9.49

PDGF-AA NA 55.77 202.14 53.72 61.89 61.12 51.57 370.6 74.34 23.48

PDGF-

AB/BB

NA 0 0 0 0 0 0 0 0 0

RANTES NA 0 31.26 35.13 202.69 106.27 10.65 0 69.44 4.71

sCD-40L NA 1.88 2.92 0.73 4.47 2.21 0.99 0.73 4.07 2.56

sIL-2Ra NA 0 0.74 0 10.45 2.01 0 0 4.35 1.2

TGF-a NA 0 12.11 0 8.63 0 8.27 0 3.16 4.96

TNF-a NA 0 0 0 0.45 0 0 0 0 0

TNF-b NA 0 0.73 0 0 0 0 0 0 0

VEGF NA 0 50.92 0 176.64 6.68 0 0 21.28 32.91

Suppl Table 2:

Gene D1 D3 D2 N2 N3 N1

CLEC3B 1.0E+06 5.5E+06 3.1E+06 5.0E+01 5.0E+01 5.0E+01

ATP5A1 1.1E+06 2.9E+06 4.4E+06 5.0E+01 5.0E+01 5.0E+01

KRT18 1.5E+06 1.2E+06 3.0E+06 5.0E+01 5.0E+01 5.0E+01

CAPZA1 7.4E+05 1.4E+06 2.8E+06 5.0E+01 5.0E+01 5.0E+01

LOC100133511 7.3E+05 1.3E+06 2.4E+06 5.0E+01 5.0E+01 5.0E+01

FETUB 1.2E+05 1.7E+06 2.5E+06 5.0E+01 5.0E+01 5.0E+01

HSPD1 5.0E+01 7.5E+05 3.5E+06 5.0E+01 5.0E+01 5.0E+01

GPR126 1.5E+05 2.5E+06 5.8E+05 5.0E+01 5.0E+01 5.0E+01

TPM1 7.7E+04 5.7E+05 1.5E+06 2.0E+01 5.0E+01 5.0E+01

SUCLG2 2.7E+05 9.6E+05 1.9E+06 5.0E+01 1.1E+02 5.0E+01

TTBK2 8.8E+04 2.8E+05 1.9E+06 5.0E+01 5.0E+01 5.0E+01

SERPINA10 5.0E+01 1.4E+06 7.0E+05 5.0E+01 5.0E+01 5.0E+01

ATP1A2 5.0E+01 4.1E+05 2.6E+06 5.0E+01 1.5E+02 5.0E+01

LUM 5.0E+01 1.2E+06 6.2E+05 5.0E+01 5.0E+01 5.0E+01

BCAS1 6.9E+04 6.9E+05 1.1E+06 5.0E+01 5.0E+01 5.0E+01

HPGD 5.0E+01 4.0E+05 1.4E+06 5.0E+01 5.0E+01 5.0E+01


15

ATP6V1E1 4.2E+05 4.9E+05 8.8E+05 5.0E+01 5.0E+01 5.0E+01

AKR1D1 2.6E+05 1.1E+06 3.5E+05 5.0E+01 5.0E+01 5.0E+01

MUPCDH 5.0E+01 4.4E+05 1.2E+06 5.0E+01 5.0E+01 5.0E+01

EFNA1 5.0E+01 1.1E+06 5.3E+05 5.0E+01 5.0E+01 5.0E+01

CYB5A 5.5E+05 4.3E+05 6.2E+05 5.0E+01 5.0E+01 5.0E+01

CRNN 3.3E+05 6.1E+05 6.3E+05 5.0E+01 5.0E+01 5.0E+01

TRAP1 7.2E+04 5.6E+05 8.3E+05 5.0E+01 5.0E+01 5.0E+01

IGLC1;IGLV1-

44;IGLV1-40;IGLV3-

21;IGLV2-11;IGLV2-

14;IGL@;IGLC2;IGLC

3

4.2E+06 7.2E+05 5.0E+01 4.5E+02 5.0E+01 5.0E+01

APOL1 5.0E+01 1.2E+06 9.0E+04 5.0E+01 5.0E+01 5.0E+01

HIBADH 2.9E+05 3.2E+05 7.0E+05 5.0E+01 5.0E+01 5.0E+01

COL1A1 2.0E+05 7.7E+05 2.7E+05 5.0E+01 5.0E+01 5.0E+01

IPI00386135 1.7E+06 5.0E+01 4.1E+03 1.3E+02 5.0E+01 5.0E+01

AK3 2.6E+05 3.6E+05 5.1E+05 5.0E+01 5.0E+01 5.0E+01

ARHGDIA 2.3E+05 2.9E+05 5.8E+05 5.0E+01 5.0E+01 5.0E+01

ATP5B 1.0E+06 1.4E+06 1.4E+06 5.0E+01 4.4E+02 5.0E+01

EPS8 1.1E+05 4.0E+05 5.5E+05 5.0E+01 5.0E+01 5.0E+01

COPZ1 1.9E+05 1.9E+05 6.8E+05 5.0E+01 5.0E+01 5.0E+01

PSME1 1.8E+05 4.6E+05 4.1E+05 5.0E+01 5.0E+01 5.0E+01

PTGR1 1.1E+05 4.0E+05 5.1E+05 5.0E+01 5.0E+01 5.0E+01

DSC2 1.7E+05 2.3E+05 1.0E+06 5.0E+01 1.3E+02 5.0E+01

SIRPA 1.2E+05 8.2E+05 6.2E+02 5.0E+01 5.0E+01 5.0E+01

ARPC5 1.6E+05 3.8E+05 3.7E+05 5.0E+01 5.0E+01 5.0E+01

NUCB2 2.2E+05 1.5E+05 4.8E+05 5.0E+01 5.0E+01 5.0E+01

PBLD 5.0E+01 2.3E+05 6.2E+05 5.0E+01 5.0E+01 5.0E+01

SULT1A3;SULT1A4 5.0E+01 2.6E+05 5.7E+05 5.0E+01 5.0E+01 5.0E+01

APOF 5.0E+01 5.7E+05 2.4E+05 5.0E+01 5.0E+01 5.0E+01

HEPH 1.8E+05 1.3E+05 3.7E+05 5.0E+01 5.0E+01 5.0E+01

HSPA9 3.8E+05 2.7E+05 1.9E+03 5.0E+01 5.0E+01 5.0E+01

DDX3X 9.9E+04 1.1E+05 4.2E+05 5.0E+01 5.0E+01 5.0E+01

RPL17;LOC1001339

31

5.0E+01 1.6E+05 4.6E+05 5.0E+01 5.0E+01 5.0E+01

DGAT1 1.3E+05 2.3E+05 2.5E+05 5.0E+01 5.0E+01 5.0E+01

COPB1 5.0E+01 1.9E+05 4.1E+05 5.0E+01 5.0E+01 5.0E+01

PGLYRP2 6.4E+04 3.4E+05 1.9E+05 5.0E+01 5.0E+01 5.0E+01

ARPC2 5.0E+01 1.9E+05 3.6E+05 5.0E+01 5.0E+01 5.0E+01

DBI 1.8E+05 2.1E+05 1.4E+05 5.0E+01 5.0E+01 5.0E+01

IGHG1;IGHG2;IGHV4

-31;IGH@

5.2E+05 5.0E+01 6.6E+04 5.0E+01 7.4E+01 5.0E+01

MYH14 2.9E+05 2.3E+05 6.4E+05 5.0E+01 2.7E+02 5.0E+01

IGLV10-54 1.6E+05 3.1E+05 8.1E+02 5.0E+01 5.0E+01 5.0E+01

DCI 2.4E+05 2.2E+05 3.3E+05 5.0E+01 1.7E+02 5.0E+01


16

MYL1 9.6E+04 1.7E+05 1.7E+05 5.0E+01 5.0E+01 5.0E+01

KIAA1967 5.0E+01 1.5E+05 2.7E+05 5.0E+01 5.0E+01 5.0E+01

IPI00384394 4.2E+05 5.0E+01 2.1E+03 5.0E+01 5.0E+01 5.0E+01

SLC16A1 5.0E+01 1.1E+05 3.1E+05 5.0E+01 5.0E+01 5.0E+01

UGT2B28 4.3E+05 5.0E+01 1.2E+05 5.0E+01 1.4E+02 5.0E+01

ARPC4;TTLL3 5.0E+01 1.0E+05 2.4E+05 5.0E+01 5.0E+01 5.0E+01

RPLP2 7.9E+05 2.2E+06 6.4E+06 4.1E+03 5.0E+01 5.0E+01

SKP1 7.0E+04 1.3E+05 7.4E+04 5.0E+01 5.0E+01 5.0E+01

RPLP0 3.7E+05 1.2E+06 1.9E+06 2.1E+03 5.0E+01 5.0E+01

PSMD2 5.0E+01 1.2E+05 1.2E+05 5.0E+01 5.0E+01 5.0E+01

ACAA1 5.0E+01 8.4E+04 2.8E+05 5.0E+01 1.9E+02 5.0E+01

PHB 5.5E+05 6.8E+05 1.2E+06 5.0E+01 5.0E+01 1.9E+03

IPI00017870 1.7E+06 1.8E+06 5.0E+06 7.1E+03 5.0E+01 5.0E+01

LIPF 1.8E+05 5.0E+01 2.9E+05 5.0E+01 3.4E+02 5.0E+01

RAB14 1.3E+05 5.0E+01 2.8E+05 5.0E+01 3.3E+02 5.0E+01

HNRNPK 1.2E+05 5.0E+01 1.8E+05 5.0E+01 2.1E+02 5.0E+01

RPN1 1.2E+05 5.0E+01 2.0E+05 5.0E+01 2.4E+02 5.0E+01

MARCKSL1 5.0E+01 5.6E+04 8.1E+04 5.0E+01 5.0E+01 5.0E+01

DDAH1 9.9E+04 5.0E+01 1.9E+05 5.0E+01 2.2E+02 5.0E+01

SPTAN1 8.9E+04 5.0E+01 9.4E+04 5.0E+01 1.1E+02 5.0E+01

SPRR2C;SPRR2G 1.3E+05 5.0E+01 6.3E+02 5.0E+01 5.0E+01 5.0E+01

RPLP1 5.0E+01 5.0E+01 2.8E+06 5.0E+01 3.3E+03 5.0E+01

ALPI 7.0E+04 5.0E+01 2.3E+05 5.0E+01 2.7E+02 5.0E+01

AK2 2.0E+06 2.1E+06 1.9E+06 5.0E+01 5.0E+01 7.6E+03

ALDH3A2 6.4E+04 5.0E+01 1.2E+05 5.0E+01 1.4E+02 5.0E+01

IGLV3-25 5.0E+01 5.0E+01 5.7E+05 5.0E+01 6.7E+02 5.0E+01

SFN 5.0E+01 5.0E+01 5.5E+05 5.0E+01 6.5E+02 5.0E+01

ICAM1 5.0E+01 1.0E+05 5.0E+01 5.0E+01 5.0E+01 5.0E+01

ELANE 5.0E+01 5.0E+01 3.0E+05 5.0E+01 3.5E+02 5.0E+01

EPB41L3 8.0E+05 1.4E+06 2.4E+06 5.0E+01 5.0E+01 6.9E+03

LOC723972;ANP32A 5.0E+01 9.6E+04 5.0E+01 5.0E+01 5.0E+01 5.0E+01

TXNDC17 5.0E+01 5.0E+01 2.6E+05 5.0E+01 3.0E+02 5.0E+01

PGA3;PGA4;PGA5 5.0E+01 5.0E+01 2.5E+05 5.0E+01 3.0E+02 5.0E+01

ITIH2 3.1E+06 1.5E+07 2.0E+07 3.4E+03 7.3E+02 5.7E+04

GPD1 5.0E+01 5.0E+01 1.7E+05 5.0E+01 2.0E+02 5.0E+01

UBE2V1;TMEM189-

UBE2V1

5.0E+01 5.0E+01 1.6E+05 5.0E+01 1.8E+02 5.0E+01

SERPINF1 1.4E+06 4.0E+06 3.8E+06 5.0E+01 5.0E+01 1.7E+04

GSTT1 5.0E+01 7.9E+04 5.0E+01 5.0E+01 5.0E+01 5.0E+01

MEP1B 5.0E+01 5.0E+01 1.0E+05 5.0E+01 1.2E+02 5.0E+01

CHGA 5.0E+01 7.0E+04 5.0E+01 5.0E+01 5.0E+01 5.0E+01

RANP1;RAN 5.0E+01 6.6E+04 5.0E+01 5.0E+01 5.0E+01 5.0E+01

YWHAH 5.0E+01 5.0E+01 8.3E+04 5.0E+01 9.8E+01 5.0E+01

YWHAQ 5.0E+01 5.0E+01 7.2E+04 5.0E+01 8.5E+01 5.0E+01

HIST2H2BE 3.6E+05 8.5E+04 1.7E+06 4.8E+03 4.9E+02 5.0E+01


17

C9 1.3E+06 1.2E+07 5.7E+06 1.2E+04 5.0E+01 4.6E+04

HSP90AA1 1.1E+06 1.7E+06 2.2E+06 5.0E+01 5.0E+01 2.0E+04

APOC1 3.9E+05 4.6E+06 2.6E+06 5.0E+01 5.0E+01 3.0E+04

AGR2 8.5E+05 7.5E+05 1.1E+06 1.1E+04 5.0E+01 5.0E+01

AHSG 1.1E+07 2.6E+07 2.0E+07 1.0E+04 1.0E+05 1.2E+05

EIF5A2 2.3E+05 2.7E+06 5.2E+06 5.0E+01 5.0E+01 3.4E+04

FBP1 9.0E+05 1.9E+06 2.1E+06 5.0E+01 5.0E+01 2.5E+04

YWHAZ 6.8E+05 4.1E+05 5.7E+05 6.6E+01 6.6E+02 8.3E+03

PKM2 4.5E+05 2.3E+06 2.4E+06 5.0E+01 2.9E+04 5.0E+01

HIST1H2AB;HIST1H

2AE;HIST1H2AL;HIS

T1H2AG;HIST1H2AJ;

HIST1H2AK;HIST1H

2AI;HIST1H2AM

1.8E+05 4.4E+05 5.2E+06 5.0E+01 5.0E+01 3.5E+04

HIST1H4J;HIST1H4E

;HIST1H4H;HIST1H

4K;HIST2H4A;HIST1

H4D;HIST1H4F;HIST

2H4B;HIST1H4C;HIS

T1H4B;HIST1H4I;HI

ST1H4L;HIST1H4A;

HIST4H4

4.5E+06 2.3E+06 9.7E+06 5.8E+02 2.4E+03 1.1E+05

AFM 1.7E+06 5.3E+06 3.7E+06 5.0E+01 5.0E+01 7.8E+04

IDH1 1.5E+06 2.3E+06 2.2E+06 5.0E+01 5.0E+01 4.5E+04

ETFB 1.1E+06 1.8E+06 2.3E+06 4.1E+03 5.0E+01 4.2E+04

PFN1 3.4E+06 3.1E+06 7.4E+06 5.0E+01 5.0E+01 1.3E+05

MYL6;MYL6B 5.0E+01 2.9E+05 1.1E+06 1.3E+04 5.0E+01 5.0E+01

CFL1 3.2E+05 2.1E+06 3.4E+06 5.0E+01 2.2E+02 5.5E+04

FABP2 4.4E+05 5.0E+01 7.3E+04 5.0E+01 9.0E+01 4.7E+03

APOA1 1.1E+05 7.6E+07 5.1E+07 5.2E+05 3.5E+05 4.0E+05

KRT19 6.0E+05 1.0E+06 3.2E+06 1.4E+04 1.3E+02 4.1E+04

CMAS 2.1E+05 2.1E+05 8.9E+05 5.0E+01 5.0E+01 1.7E+04

MYH14 3.8E+05 6.7E+05 4.0E+06 5.0E+01 9.2E+02 6.6E+04

PURA 9.4E+04 5.0E+01 1.6E+06 5.0E+01 1.9E+03 2.1E+04

HGFAC 6.5E+04 3.0E+06 1.0E+06 5.0E+01 5.6E+04 5.0E+01

ACSL5 2.5E+05 6.5E+05 7.1E+05 5.0E+01 2.6E+02 2.2E+04

IPI00398129 2.2E+05 5.8E+05 1.2E+06 2.7E+03 5.0E+01 2.7E+04

HIST2H3D;HIST2H3

C;HIST2H3A

6.6E+05 9.4E+04 4.9E+06 2.3E+04 1.3E+02 6.0E+04

BTD 1.5E+06 5.9E+06 5.0E+06 5.0E+01 1.0E+04 1.7E+05

UQCRC2 5.7E+06 4.0E+06 9.8E+06 5.0E+01 5.0E+01 2.9E+05

CPS1 8.1E+05 3.7E+06 4.6E+06 1.0E+02 3.8E+04 1.0E+05

ANXA13 2.3E+05 2.1E+06 6.5E+06 5.0E+01 1.2E+05 3.1E+04

HNRNPL 1.3E+06 5.8E+05 1.5E+06 1.3E+04 5.0E+01 4.7E+04

PROC 5.9E+04 1.2E+06 6.3E+05 5.0E+01 5.0E+01 3.4E+04

APOA4 9.0E+05 7.0E+07 3.5E+07 1.2E+06 4.0E+05 4.4E+05


18

TST 2.0E+05 2.5E+05 1.3E+06 5.0E+01 6.2E+02 3.4E+04

HRG 6.1E+06 9.4E+06 6.8E+06 4.5E+04 2.8E+05 1.3E+05

NP 1.7E+06 1.6E+06 3.1E+06 1.1E+05 5.0E+01 2.0E+04

LMNA 7.2E+05 5.1E+05 1.2E+06 9.6E+03 5.0E+01 4.3E+04

DEFA5 3.8E+06 3.2E+06 7.0E+06 1.2E+02 1.4E+05 1.7E+05

CNDP1 2.3E+05 1.7E+06 9.5E+05 5.0E+01 2.9E+04 4.6E+04

MYH9 1.8E+06 3.7E+06 1.6E+07 4.5E+05 7.1E+04 4.3E+04

SLC9A3R1 2.7E+06 4.4E+06 3.4E+06 2.0E+04 1.5E+05 1.2E+05

HYOU1 1.2E+06 1.4E+06 1.6E+06 5.0E+01 5.0E+01 1.2E+05

HBG1;HBG2 2.2E+05 8.1E+06 3.6E+06 5.0E+01 1.0E+05 2.4E+05

PRDX5 9.6E+05 1.2E+06 1.2E+06 2.0E+04 5.0E+01 7.8E+04

KRT8 5.8E+06 5.5E+06 1.9E+07 1.3E+05 3.0E+05 4.4E+05

EEF1A1 5.0E+01 5.0E+05 1.5E+06 5.9E+04 5.0E+01 5.0E+01

ACO2 1.2E+06 1.3E+06 2.3E+06 5.0E+01 3.1E+02 1.4E+05

GKN1 4.7E+04 1.4E+02 4.0E+06 8.6E+04 3.5E+04 5.0E+01

HSPA7 6.3E+05 7.6E+05 1.2E+06 5.0E+01 5.0E+01 7.9E+04

AKR7A3 3.0E+05 1.1E+06 1.1E+06 5.3E+04 5.0E+01 2.3E+04

APOC3 5.0E+01 1.4E+06 6.6E+05 1.0E+04 5.0E+01 5.3E+04

REG4 1.7E+05 7.9E+05 2.8E+06 6.5E+04 5.7E+04 5.0E+01

ACAT1 2.2E+06 2.1E+06 4.5E+06 5.0E+01 7.9E+04 2.0E+05

UQCRB 7.1E+05 6.0E+05 9.7E+05 5.0E+01 5.0E+01 7.5E+04

ITIH3 8.8E+05 2.8E+06 3.2E+06 1.8E+02 5.0E+01 2.3E+05

GLUD1 2.6E+06 1.5E+06 1.7E+06 5.0E+01 5.0E+01 2.0E+05

PGC 1.4E+07 1.5E+06 1.0E+07 2.0E+05 4.0E+05 2.8E+05

AGT 3.0E+06 9.5E+06 5.9E+06 1.5E+04 9.5E+01 6.4E+05

CKMT1B;LOC10013

3623;CKMT1A

5.0E+01 1.2E+06 2.1E+06 6.0E+04 5.0E+01 5.7E+04

APOA2 6.0E+04 6.5E+06 3.6E+06 2.8E+04 5.1E+04 2.9E+05

IPI00382440 6.7E+05 8.4E+05 1.2E+06 5.0E+01 5.0E+01 1.0E+05

PCDH24 2.0E+05 2.8E+06 5.0E+05 5.0E+01 1.3E+05 5.0E+01

PDIA3 6.4E+06 4.0E+06 2.5E+06 2.6E+04 5.0E+01 4.8E+05

C8B 2.7E+05 1.9E+06 6.3E+05 5.0E+01 1.1E+05 5.0E+01

CFI 5.7E+06 1.9E+07 1.0E+07 1.2E+05 7.6E+05 5.2E+05

IGFALS 3.2E+05 5.2E+05 4.1E+05 5.0E+01 5.2E+04 5.0E+01

ARCN1 8.6E+05 1.1E+06 1.0E+06 7.6E+04 5.0E+01 5.9E+04

PRDX3 1.4E+06 1.0E+06 2.2E+06 1.6E+03 6.3E+04 1.5E+05

IPI00382489 2.9E+06 7.8E+05 2.8E+05 5.0E+01 4.9E+04 1.3E+05

VNN1 1.7E+05 9.4E+05 7.6E+05 5.0E+01 8.8E+04 5.0E+01

SLC5A1 2.5E+05 3.8E+05 3.5E+05 5.0E+01 5.0E+01 4.7E+04

ETFA 2.9E+06 2.9E+06 1.0E+06 8.5E+03 6.2E+04 2.6E+05

HSP90B1 2.9E+06 3.2E+06 3.7E+06 5.0E+01 2.5E+05 2.2E+05

CD59 6.8E+06 6.5E+06 3.3E+06 5.3E+03 2.9E+05 5.3E+05

IPI00782983 6.9E+05 1.1E+06 8.6E+04 5.0E+01 5.0E+01 9.2E+04

OAT 1.1E+06 1.5E+06 2.7E+06 5.0E+01 1.8E+05 8.6E+04

PCK2 7.5E+05 7.8E+05 7.9E+05 5.0E+01 6.8E+04 5.2E+04


19

IGJ 3.5E+07 1.0E+07 3.3E+07 2.4E+05 1.9E+06 2.1E+06

MYO1A 2.9E+05 1.9E+05 1.6E+05 5.0E+01 5.0E+01 3.5E+04

LCT 2.8E+05 7.3E+05 2.4E+06 1.5E+04 8.0E+04 9.6E+04

UQCRC1 4.7E+05 7.6E+05 1.8E+06 5.0E+01 1.1E+05 6.4E+04

RAB35 3.7E+05 2.2E+05 8.3E+05 8.1E+04 2.8E+02 5.0E+01

IPI00829624 9.5E+05 3.3E+05 2.3E+05 5.0E+01 5.0E+01 8.6E+04

B2M 6.0E+05 1.4E+05 1.1E+05 3.4E+04 5.0E+01 1.5E+04

KRT4 2.6E+06 1.6E+06 8.9E+06 2.8E+05 3.7E+05 1.3E+05

PPA1 1.5E+06 9.2E+05 1.9E+06 1.4E+05 1.1E+03 1.2E+05

CPN2 1.7E+05 5.0E+05 2.3E+05 5.0E+01 1.3E+02 5.4E+04

IPI00003469 6.8E+05 7.7E+05 5.0E+01 5.0E+01 3.8E+04 5.1E+04

PRB3 4.4E+05 3.1E+02 1.4E+06 8.7E+04 6.1E+02 2.5E+04

NPC2 4.2E+05 2.7E+05 4.8E+05 1.9E+04 5.0E+01 5.5E+04

KNG1 9.8E+06 2.9E+07 2.1E+07 3.2E+05 1.7E+06 1.8E+06

PRSS8 2.3E+05 3.7E+05 4.9E+05 5.1E+02 3.9E+04 3.1E+04

EEF2 1.4E+06 3.0E+06 5.9E+06 3.3E+05 1.9E+05 1.5E+05

ITIH1 9.6E+06 1.6E+07 1.8E+07 5.5E+03 2.6E+05 2.7E+06

ACTN2 7.0E+05 1.1E+02 4.4E+05 2.6E+04 5.0E+01 5.3E+04

PGA3;PGA4;PGA5 1.2E+07 2.5E+06 2.7E+07 1.9E+06 2.3E+05 8.7E+05

ALDOB 1.3E+07 1.4E+07 1.8E+07 7.7E+05 1.2E+06 1.3E+06

TUBA4A 1.9E+05 7.7E+05 1.5E+06 2.7E+02 2.5E+02 1.7E+05

PRDX2 1.9E+06 3.7E+06 6.3E+06 1.9E+05 3.3E+04 6.5E+05

TSPAN8 2.1E+05 9.7E+04 8.3E+05 3.1E+04 6.5E+02 5.2E+04

SERPINC1 5.4E+06 2.8E+07 3.2E+07 4.2E+05 1.8E+06 2.7E+06

CLIC1 2.2E+06 2.0E+06 3.4E+06 4.6E+04 1.1E+05 4.0E+05

PGD 2.3E+05 2.5E+05 4.5E+05 5.0E+01 7.1E+04 5.0E+01

RDX 7.6E+05 2.9E+05 9.7E+05 5.0E+01 5.0E+01 1.5E+05

C6 7.7E+05 2.0E+06 2.3E+06 1.0E+05 1.1E+05 1.9E+05

TXN 6.7E+06 6.2E+06 2.0E+07 7.1E+05 4.1E+05 1.5E+06

MANF 1.9E+05 5.0E+01 2.2E+05 5.0E+01 2.6E+02 3.1E+04

HABP2 4.1E+05 5.0E+01 5.0E+01 1.7E+02 5.0E+01 3.3E+04

FBLN1 4.8E+05 7.8E+05 1.1E+05 5.0E+01 5.0E+01 1.1E+05

NT5E 8.9E+04 1.8E+06 1.2E+06 8.5E+04 1.8E+05 5.0E+01

QSOX1 1.7E+06 6.4E+05 3.3E+06 7.1E+04 1.8E+05 2.3E+05

C7 1.7E+06 4.2E+06 2.5E+06 1.4E+05 3.0E+05 2.9E+05

PON1 1.7E+06 4.6E+06 4.3E+06 1.2E+05 3.1E+05 5.1E+05

IPI00386136 1.2E+06 5.0E+01 6.1E+03 2.9E+02 5.0E+01 1.1E+05

C8A 1.3E+06 2.0E+06 1.7E+05 1.8E+02 6.0E+04 2.6E+05

KRT13 1.4E+06 3.4E+05 2.1E+06 1.0E+05 1.4E+05 1.2E+05

SLC25A5 3.7E+06 6.5E+06 6.6E+06 3.4E+04 4.8E+05 1.1E+06

HSP90AB1 8.3E+05 8.4E+05 1.4E+06 5.0E+01 6.5E+04 2.2E+05

ALPP 1.1E+05 1.0E+05 3.0E+05 4.9E+04 5.0E+01 5.0E+01

F2 5.1E+06 1.2E+07 8.5E+06 8.6E+05 6.0E+05 1.0E+06

IPI00783024 9.0E+05 2.1E+06 1.6E+05 1.4E+02 2.3E+04 2.8E+05

EPS8L3 4.3E+05 3.8E+05 8.9E+05 5.0E+01 3.1E+04 1.3E+05


20

PDCD6IP 2.8E+05 2.0E+05 3.7E+05 5.0E+01 5.0E+01 8.3E+04

CST3 5.9E+06 2.1E+06 2.8E+06 7.1E+05 3.3E+05 5.8E+04

GSN 5.8E+06 1.0E+07 1.1E+07 1.1E+06 4.3E+05 1.4E+06

HNRNPA2B1 8.1E+05 4.3E+05 2.3E+06 9.3E+04 1.9E+05 9.3E+04

FGA 1.4E+07 2.1E+07 2.5E+07 1.5E+05 3.0E+06 3.3E+06

HSPA1A;HSPA1B 6.4E+04 5.0E+01 2.0E+06 5.0E+01 5.1E+04 1.7E+05

HSD17B2 2.2E+06 2.3E+06 1.5E+06 5.0E+01 4.5E+04 6.2E+05

SDHB 4.6E+05 5.0E+01 1.0E+06 1.7E+04 9.7E+04 4.9E+04

RHOC 3.0E+05 3.2E+05 4.0E+05 5.0E+01 5.0E+01 1.2E+05

PPIAP19 1.9E+06 7.2E+05 1.9E+06 5.0E+01 1.0E+05 4.4E+05

IGHG1;IGHG2;IGHV4

-31;IGH@

4.9E+05 7.9E+04 2.8E+05 3.5E+02 5.0E+01 1.0E+05

GSTA3 5.0E+06 3.2E+06 2.9E+06 8.3E+04 3.1E+05 9.6E+05

F11R 4.7E+05 7.5E+05 1.7E+06 5.0E+01 1.1E+05 2.6E+05

IPI00019591 8.3E+06 2.4E+07 1.4E+07 4.0E+02 4.0E+05 5.8E+06

HADHA 2.7E+06 3.2E+06 4.0E+06 4.8E+05 2.0E+05 6.5E+05

MDH1 4.7E+05 5.0E+05 1.1E+05 2.4E+02 5.0E+01 1.5E+05

MUC13 4.1E+06 1.9E+06 5.4E+06 1.5E+05 3.4E+05 1.1E+06

CD36 1.6E+06 9.7E+05 2.5E+06 1.1E+04 3.6E+05 3.4E+05

KRT20 1.6E+06 1.7E+06 6.0E+06 2.1E+02 7.1E+04 1.2E+06

IPI00924820 5.9E+05 2.7E+05 6.4E+05 5.0E+01 3.6E+04 1.7E+05

IPI00164838 2.0E+05 2.5E+05 1.0E+05 5.0E+01 5.0E+01 7.6E+04

GSTA5 8.2E+05 5.0E+01 4.4E+05 5.0E+01 5.5E+02 1.8E+05

SOD1 5.9E+05 8.7E+05 3.2E+05 2.1E+05 4.7E+04 5.0E+01

ANXA2 2.2E+06 2.6E+06 4.2E+06 3.8E+04 5.4E+05 7.3E+05

CTSB 1.4E+05 4.1E+05 1.4E+05 5.0E+01 1.0E+05 5.0E+01

OTC 7.6E+05 5.6E+05 6.7E+05 9.8E+04 6.9E+04 1.3E+05

LRG1 1.5E+06 2.7E+06 1.3E+06 5.0E+01 2.3E+05 6.0E+05

CES2 7.9E+05 1.2E+06 1.2E+06 5.0E+01 2.2E+05 2.6E+05

LIPF 6.6E+07 4.0E+06 5.3E+07 1.4E+07 3.8E+06 1.0E+06

ACTA2 1.2E+07 1.6E+07 1.8E+07 6.5E+05 2.3E+06 4.1E+06

OLFM4 1.1E+07 1.1E+06 3.4E+06 8.4E+04 1.2E+06 1.2E+06

ACE2 1.4E+06 1.2E+06 4.2E+06 1.1E+05 9.0E+05 4.8E+04

APOE 2.4E+05 6.4E+06 2.2E+06 9.0E+05 3.3E+05 1.8E+05

PPIB 9.4E+05 7.5E+05 1.2E+06 3.4E+04 1.1E+02 4.5E+05

ECHS1 5.2E+05 1.0E+06 1.9E+06 5.0E+01 1.9E+05 3.8E+05

HPGD 2.0E+05 3.3E+05 5.8E+05 9.7E+04 3.3E+04 5.3E+04

MSMB 1.3E+05 1.0E+05 2.2E+05 7.5E+04 5.0E+01 5.0E+01

HMGCS2 2.8E+05 7.4E+05 4.0E+05 5.7E+04 5.2E+04 1.3E+05

LYZ 4.4E+07 1.5E+07 2.6E+07 6.3E+06 2.7E+06 5.7E+06

MAOA 3.2E+05 2.6E+05 6.8E+05 5.0E+01 2.1E+02 2.2E+05

GPA33 6.3E+05 4.0E+05 1.6E+06 8.5E+04 2.3E+05 1.5E+05

SUCLG1 1.1E+06 1.2E+06 1.3E+06 5.0E+01 4.8E+05 1.6E+05

PIGR 1.0E+08 4.3E+07 7.3E+07 1.4E+07 1.3E+07 1.2E+07

ATP1B3 3.7E+05 2.4E+05 3.3E+05 5.0E+01 7.4E+04 9.4E+04


21

FABP5L9;FABP5L2;

FABP5;FABP5L7

5.0E+01 7.4E+01 1.3E+05 2.4E+04 5.0E+01 5.0E+01

CRISP3 6.1E+05 1.0E+05 1.9E+05 3.5E+04 1.2E+02 1.3E+05

GOLM1 1.5E+06 3.5E+05 3.6E+06 4.7E+05 1.6E+05 3.7E+05

FAM3C 5.8E+05 1.7E+05 1.1E+06 3.9E+04 1.3E+05 1.7E+05

ALDH2 3.9E+06 2.1E+06 3.0E+06 1.6E+05 7.4E+05 7.7E+05

ACAA2 4.8E+05 2.3E+05 6.9E+05 1.3E+05 9.0E+04 4.1E+04

FCN3 6.5E+05 2.0E+06 1.5E+06 5.0E+01 2.6E+05 5.3E+05

PLG 1.5E+07 2.2E+07 1.3E+07 5.5E+05 3.0E+06 5.7E+06

CLU 3.6E+06 2.0E+07 1.8E+07 4.8E+05 5.0E+06 2.5E+06

IGHM 5.6E+07 1.8E+07 2.4E+07 1.1E+06 7.1E+06 1.1E+07

FABP1 1.2E+07 1.1E+07 1.6E+07 8.1E+05 1.9E+06 5.0E+06

GC 2.1E+07 2.7E+07 2.1E+07 2.3E+05 3.6E+06 9.9E+06

GPLD1 6.4E+05 1.2E+06 1.2E+06 5.0E+01 5.0E+01 6.2E+05

CLTC 1.5E+05 1.1E+06 1.1E+06 1.5E+05 1.8E+05 1.5E+05

TUBB 8.0E+04 5.0E+01 9.7E+04 5.0E+01 1.2E+02 3.6E+04

APOB 2.1E+07 6.4E+07 8.2E+07 4.0E+06 6.1E+06 2.4E+07

PRDX1 7.3E+06 7.0E+06 1.2E+07 7.0E+05 3.1E+06 1.6E+06

YWHAE 1.1E+06 5.0E+01 3.2E+05 5.0E+01 4.3E+02 2.8E+05

MASP2 4.1E+05 1.4E+05 1.0E+05 5.0E+01 5.0E+01 1.4E+05

HEPACAM 5.0E+01 3.9E+05 5.7E+04 5.9E+04 3.5E+04 5.0E+01

KLKB1 1.9E+06 4.5E+06 1.1E+06 6.0E+05 3.4E+05 6.7E+05

IGHA1 1.8E+06 1.3E+06 2.5E+02 1.1E+02 2.2E+05 4.5E+05

SERPINA7 7.7E+05 6.1E+05 6.5E+05 5.0E+01 7.8E+04 3.7E+05

ALB 3.0E+06 3.1E+06 1.2E+06 5.0E+01 4.0E+04 1.6E+06

CFHR2 1.7E+06 4.8E+06 1.7E+06 5.0E+01 4.1E+05 1.4E+06

ANPEP 2.6E+07 1.1E+07 1.3E+07 4.3E+06 5.5E+06 1.7E+06

GIF 5.6E+05 5.0E+01 1.3E+05 1.3E+05 3.0E+04 5.0E+01

UBC;RPS27A;UBB 2.2E+06 1.6E+06 5.0E+06 1.0E+06 4.6E+05 4.9E+05

ATRN 1.2E+06 1.3E+06 7.5E+05 2.5E+04 2.1E+05 5.2E+05

C8G 1.2E+06 1.8E+06 1.1E+06 2.3E+04 1.6E+05 7.6E+05

IPI00827815 2.2E+06 1.2E+06 3.6E+06 5.0E+01 4.1E+05 1.2E+06

SPINK1 4.3E+05 6.1E+05 7.2E+05 5.0E+01 2.0E+05 2.0E+05

SI 9.8E+06 4.5E+06 1.2E+07 7.3E+05 3.2E+06 2.1E+06

IPI00386137 5.0E+01 3.8E+05 4.4E+05 5.0E+01 8.5E+04 1.0E+05

CDH17 3.7E+06 3.4E+06 1.1E+07 9.8E+05 2.0E+06 1.3E+06

MVP 4.4E+05 7.2E+05 2.6E+06 2.6E+05 4.8E+05 2.0E+05

HSP90AA2 1.6E+05 5.0E+01 1.4E+05 2.1E+02 1.6E+02 7.2E+04

APOD 1.9E+06 2.5E+06 1.9E+06 1.1E+05 3.8E+05 1.0E+06

MUC6 1.1E+08 1.6E+07 6.2E+07 6.6E+06 2.0E+07 2.1E+07

HLA-H 1.3E+05 7.5E+04 5.0E+01 1.5E+02 5.0E+01 5.2E+04

S100A9 2.4E+05 6.3E+05 7.6E+05 1.5E+05 7.1E+04 1.9E+05

LOC100126583;IGH

A2

9.0E+05 4.2E+05 4.8E+05 5.0E+01 9.1E+04 3.7E+05

SERPIND1 2.7E+06 7.0E+06 6.7E+06 6.0E+02 1.1E+06 3.2E+06


22

PTGR1 5.5E+05 8.4E+05 7.0E+05 3.3E+05 1.4E+05 7.9E+04

GDI2 7.8E+05 1.7E+06 3.6E+06 2.7E+05 9.6E+05 3.7E+05

TUBA1C 1.5E+06 1.3E+06 3.4E+06 5.2E+04 5.3E+05 1.0E+06

CD5L 8.6E+06 6.0E+06 5.5E+06 7.7E+04 7.5E+05 4.5E+06

SERPINA1 1.1E+07 2.9E+07 2.9E+07 2.1E+05 6.1E+06 1.2E+07

DAK 1.3E+06 2.2E+06 1.3E+06 1.1E+06 4.7E+04 1.2E+05

SERPINA4 6.3E+05 3.4E+06 5.3E+06 9.1E+04 1.1E+06 1.3E+06

PPP1CC 2.0E+05 2.2E+05 2.6E+05 6.3E+04 7.0E+04 5.1E+04

REG3A 2.9E+06 4.5E+06 6.8E+06 9.1E+05 6.9E+05 2.2E+06

F13B 5.5E+06 3.0E+06 3.0E+06 5.0E+01 4.1E+05 2.7E+06

ACTN4 6.6E+06 4.3E+06 8.0E+06 6.3E+05 9.8E+05 3.6E+06

PDIA6 1.2E+05 5.0E+01 7.6E+04 1.7E+02 8.9E+01 5.4E+04

IPI00382439 8.1E+05 9.4E+05 1.4E+06 5.0E+01 1.5E+05 7.3E+05

IPI00384401 4.3E+06 4.0E+06 4.7E+06 4.1E+04 2.3E+05 3.4E+06

ANXA2P2 1.9E+06 1.6E+06 4.1E+06 9.8E+05 5.7E+05 6.3E+05

PARK7 5.8E+05 5.1E+05 7.1E+05 1.8E+05 5.0E+04 2.9E+05

DPP4 2.3E+05 6.7E+05 6.4E+05 3.6E+04 1.6E+05 2.5E+05

CANX 2.2E+06 1.4E+06 7.2E+06 1.4E+06 1.1E+06 6.1E+05

ACTBL2 5.8E+06 6.2E+06 4.8E+06 4.4E+05 1.8E+06 2.7E+06

ACY1 8.7E+05 2.4E+06 1.2E+06 1.3E+05 9.3E+05 2.3E+05

IPI00385264 8.6E+07 1.1E+07 1.4E+07 3.5E+06 1.2E+07 1.7E+07

IPI00830057 1.6E+05 2.3E+05 4.6E+05 5.0E+01 4.8E+04 2.0E+05

SERPING1 3.8E+06 7.6E+06 7.1E+06 1.0E+05 1.8E+06 3.5E+06

S100A8 1.7E+06 1.1E+06 5.0E+06 1.6E+06 3.5E+05 2.9E+05

ETFDH 1.8E+05 5.0E+01 1.9E+05 5.6E+04 5.2E+04 5.0E+01

P4HB 1.6E+07 9.0E+06 1.2E+07 1.6E+06 5.5E+06 3.7E+06

LOC440786 5.1E+05 5.0E+01 2.5E+05 5.0E+02 3.1E+02 2.2E+05

KRT24 1.9E+05 5.0E+01 4.5E+02 1.7E+02 5.0E+01 5.5E+04

CSTB 1.2E+06 2.8E+06 1.4E+06 1.2E+06 2.9E+05 1.6E+05

COX4I1 5.4E+05 8.0E+05 1.2E+05 3.1E+05 1.1E+05 1.8E+04

IPI00827690 3.1E+05 1.3E+05 3.4E+05 5.0E+01 3.7E+04 2.0E+05

C1R 1.7E+06 5.8E+06 4.4E+06 8.1E+04 9.9E+05 2.6E+06

IGKC 5.8E+05 5.0E+01 2.3E+05 5.0E+01 5.3E+04 2.0E+05

C1S 2.0E+06 6.3E+06 3.1E+06 5.0E+05 1.1E+06 1.9E+06

IPI00854709 3.4E+05 2.0E+05 2.7E+05 5.0E+01 5.0E+01 2.5E+05

RAB1A 1.2E+06 1.3E+06 2.3E+06 6.5E+05 3.8E+05 4.4E+05

EEF1A2 2.2E+06 2.7E+06 5.4E+06 8.7E+05 7.0E+05 1.7E+06

IGKV3D-11 1.7E+06 1.7E+06 1.4E+06 1.5E+04 3.9E+05 1.1E+06

LOC100126583;IGH

A2

4.3E+07 1.0E+07 7.6E+06 4.1E+06 8.4E+06 6.8E+06

A1BG 8.0E+06 1.6E+07 1.2E+07 1.2E+05 3.7E+06 7.7E+06

PDIA4 5.5E+05 6.3E+05 7.6E+05 1.8E+05 3.1E+05 1.4E+05

IGKV3-20 1.6E+07 1.9E+07 3.1E+06 4.5E+05 2.8E+06 9.2E+06

IGLV7-43 5.0E+05 2.3E+05 1.6E+05 5.0E+01 1.1E+05 1.8E+05

LOC100126583;IGH 1.7E+06 1.6E+06 5.0E+01 5.0E+01 3.5E+05 7.5E+05


23

A2

PGK1 2.7E+06 1.7E+06 3.2E+06 8.7E+04 7.5E+05 1.7E+06

ACTB 6.8E+06 8.0E+06 1.2E+07 1.7E+05 1.7E+06 7.1E+06

IPI00382494 5.6E+06 3.0E+06 2.2E+06 5.0E+01 9.7E+05 2.7E+06

ORM2 7.2E+06 2.1E+07 8.1E+06 1.8E+05 3.3E+06 8.9E+06

PSMB8 5.5E+05 3.9E+05 2.3E+05 5.0E+01 2.8E+05 1.4E+05

IPI00735451 3.8E+06 3.5E+06 4.7E+06 7.7E+05 8.2E+05 2.7E+06

HADHB 4.1E+06 2.9E+06 3.5E+06 9.9E+05 1.2E+06 1.6E+06

PTGDS 6.1E+05 2.2E+06 7.0E+05 5.0E+01 7.1E+05 5.5E+05

IGHA1 1.1E+07 5.6E+06 5.5E+06 1.1E+06 3.4E+06 3.5E+06

MME 1.7E+06 6.8E+05 1.6E+06 3.1E+05 6.0E+05 5.3E+05

ATP1A1 1.5E+06 9.3E+05 1.5E+06 9.6E+05 1.4E+05 3.1E+05

ACTN1 7.8E+05 1.8E+06 7.7E+05 3.4E+05 3.2E+05 5.8E+05

HSPE1 3.3E+06 1.3E+06 1.9E+06 7.9E+05 9.6E+05 6.8E+05

CAP1 1.5E+06 1.2E+06 1.9E+06 5.0E+01 9.4E+05 7.6E+05

PLS1 6.6E+06 2.9E+06 5.5E+06 2.8E+06 1.2E+06 1.6E+06

IGHG1;IGHG2;IGHV4

-31;IGH@

3.0E+06 2.4E+06 3.1E+06 6.8E+02 5.0E+05 2.7E+06

IPI00783094 3.3E+05 2.0E+05 6.4E+05 5.0E+01 1.2E+05 3.2E+05

HP 3.9E+07 1.1E+08 5.5E+07 2.6E+06 1.5E+07 5.9E+07

AKR1C4 1.9E+06 1.5E+06 1.6E+06 8.3E+05 1.3E+05 9.1E+05

IPI00382500 1.6E+06 1.8E+06 2.3E+06 3.4E+05 8.0E+05 1.0E+06

PROS1 3.9E+06 5.2E+06 3.9E+06 7.1E+04 1.5E+06 3.4E+06

SPRR1A 7.4E+05 3.4E+05 3.3E+06 1.6E+06 5.8E+04 5.0E+01

IPI00387100 2.5E+05 3.9E+05 3.6E+05 6.6E+04 1.3E+05 1.8E+05

DMBT1 4.3E+07 5.2E+06 6.7E+06 1.6E+07 3.2E+06 1.9E+06

IPI00069693 3.6E+05 5.0E+01 4.7E+05 5.0E+01 6.1E+02 3.2E+05

VIL1 5.8E+06 3.6E+06 7.3E+06 4.1E+05 1.5E+06 4.5E+06

ORM1 3.2E+06 2.4E+07 4.1E+06 5.2E+06 2.5E+06 4.2E+06

CD14 4.5E+04 2.7E+06 6.3E+05 2.4E+05 4.6E+05 6.1E+05

IPI00827944 4.5E+05 2.4E+05 1.6E+05 5.0E+01 8.4E+04 2.5E+05

PREP 3.3E+05 1.5E+05 4.2E+05 9.5E+04 1.1E+05 1.4E+05

SERPINA3 1.8E+07 1.3E+07 1.0E+07 4.4E+05 4.0E+06 1.2E+07

LOC100126583;IGH

A2

2.3E+06 8.9E+05 1.3E+05 1.7E+02 5.0E+05 8.3E+05

RBP2 1.6E+06 1.6E+06 1.7E+06 2.6E+05 1.0E+06 7.0E+05

IPI00387026 2.0E+06 6.6E+05 4.4E+05 2.1E+04 4.1E+05 8.1E+05

PPIA 5.0E+01 1.1E+02 1.1E+05 4.4E+04 1.3E+02 5.0E+01

SPRR2C;SPRR2G 4.8E+05 1.2E+02 2.0E+05 2.2E+05 4.6E+04 5.0E+01

PCK2 5.8E+06 3.4E+06 2.4E+06 1.7E+06 1.6E+06 1.3E+06

IPI00382436 4.3E+05 7.3E+05 6.8E+05 5.0E+01 8.1E+04 6.7E+05

CP 2.5E+07 4.4E+07 3.3E+07 3.3E+06 8.8E+06 3.0E+07

RAP1B 1.4E+05 7.1E+04 2.4E+05 4.8E+04 7.5E+04 5.8E+04

CES1 1.4E+05 5.0E+01 2.2E+05 5.0E+01 2.9E+02 1.5E+05

HSPA8 5.6E+06 2.2E+06 6.2E+06 1.8E+06 1.1E+06 2.9E+06


24

CFHR5 2.3E+05 2.5E+05 4.1E+05 5.0E+01 4.1E+02 3.7E+05

KRT3 6.7E+05 3.0E+05 1.5E+06 7.7E+05 2.9E+02 2.6E+05

LDHAL6A 3.2E+06 3.6E+06 6.8E+05 6.8E+03 1.6E+06 1.5E+06

IPI00384409 3.2E+05 5.0E+05 1.4E+05 5.0E+01 7.2E+04 3.2E+05

IGLL1 4.5E+06 1.3E+06 7.5E+05 3.3E+05 1.0E+06 1.4E+06

ITIH4 1.1E+07 2.0E+07 2.0E+07 1.3E+06 5.9E+06 1.4E+07

PYCARD 5.0E+01 5.0E+01 2.6E+02 5.0E+01 5.0E+01 5.0E+01

LOC284422 2.8E+05 1.3E+05 1.4E+05 5.0E+01 5.0E+01 2.3E+05

TFF1 2.0E+06 4.1E+05 1.8E+06 1.5E+06 2.4E+05 2.0E+04

CAPRIN2 1.6E+05 1.4E+05 6.0E+04 5.0E+01 4.5E+04 1.0E+05

IPI00382420 1.2E+06 8.4E+05 8.9E+04 3.8E+02 5.0E+01 9.1E+05

CPB2 2.2E+05 2.2E+05 2.7E+05 5.0E+01 1.3E+05 1.7E+05

GMDS 1.9E+05 2.9E+05 6.5E+05 1.2E+05 2.9E+05 7.3E+04

ITLN2 1.2E+06 2.0E+06 3.5E+06 1.2E+06 1.2E+06 4.9E+05

CBR1 3.8E+06 5.3E+06 9.0E+06 5.2E+06 1.5E+06 1.1E+06

ATP1B1 1.5E+06 5.0E+05 1.9E+06 4.0E+05 4.5E+05 8.6E+05

VTN 2.1E+06 1.1E+07 9.3E+06 7.6E+05 3.7E+06 5.6E+06

TP53I3 1.7E+05 5.0E+01 5.0E+01 2.3E+02 5.0E+01 7.4E+04

PRKCSH 8.9E+04 5.0E+01 1.9E+05 5.0E+01 7.4E+04 5.2E+04

EPCAM 1.6E+06 5.7E+05 1.7E+06 2.8E+04 6.4E+05 1.0E+06

VDAC1 2.8E+06 1.2E+06 8.6E+06 1.3E+06 2.5E+06 1.8E+06

CPT2 6.1E+05 1.8E+05 5.8E+05 5.0E+01 4.8E+05 1.3E+05

IGKV1-5 3.5E+05 1.6E+05 1.5E+05 5.0E+01 7.4E+04 2.2E+05

AKR1C3 9.0E+04 7.2E+05 1.5E+06 9.8E+05 5.0E+01 5.3E+04

TUBB2C 9.8E+05 4.2E+05 1.8E+06 4.0E+04 5.0E+05 9.0E+05

C5 5.1E+06 9.6E+06 8.2E+06 5.1E+05 3.6E+06 6.3E+06

FN1 1.6E+07 2.2E+07 1.4E+07 4.3E+05 4.6E+06 1.8E+07

ALDH1A1 7.1E+06 8.7E+06 1.6E+07 6.8E+06 5.5E+06 2.6E+06

ANP32B 8.8E+04 9.6E+04 3.6E+02 5.0E+01 8.7E+04 5.0E+01

GDA 6.6E+05 8.1E+05 2.5E+06 7.3E+05 5.0E+05 6.7E+05

IPI00829653 1.2E+05 6.7E+04 5.0E+01 2.5E+02 5.0E+01 9.0E+04

UGT2B17 2.5E+06 6.9E+05 3.4E+06 2.6E+06 1.5E+02 5.9E+05

IPI00829701 9.2E+05 2.5E+05 3.2E+05 1.8E+02 1.3E+05 5.8E+05

IGHA1 1.3E+08 5.4E+07 7.0E+07 2.3E+07 5.1E+07 4.7E+07

VCP 7.6E+05 4.2E+05 1.8E+06 1.4E+05 6.3E+05 6.9E+05

CEACAM5 1.4E+05 1.1E+05 1.1E+05 5.2E+04 3.4E+04 8.9E+04

GSTA1 3.2E+06 1.2E+06 2.7E+06 2.7E+06 1.2E+03 8.5E+05

EEF1G 3.1E+05 1.0E+06 2.3E+06 5.9E+05 9.9E+05 2.8E+05

LCP1 6.4E+05 2.0E+05 3.8E+05 1.3E+05 6.4E+04 4.3E+05

IGLC1;IGLV1-

44;IGLV1-40;IGLV3-

21;IGLV2-11;IGLV2-

14;IGL@;IGLC2;IGLC

3

8.6E+06 5.4E+06 5.9E+06 1.0E+05 2.9E+06 7.2E+06

FGG 7.2E+07 4.2E+07 1.8E+07 3.3E+06 1.9E+07 4.5E+07


25

ALDOA 1.7E+05 6.9E+05 4.5E+05 5.0E+01 6.2E+05 5.6E+04

LGALS3BP 6.1E+06 1.4E+06 1.1E+06 8.2E+05 2.3E+06 1.3E+06

IPI00385253 7.1E+05 3.2E+05 6.4E+05 1.2E+02 2.0E+05 6.7E+05

SDHA 9.7E+05 4.4E+05 9.3E+05 1.6E+05 9.8E+05 6.8E+04

MDH2 2.5E+06 1.1E+06 1.4E+06 1.1E+06 1.2E+06 3.5E+05

IPI00829873 2.1E+05 5.0E+01 1.3E+05 5.0E+01 1.8E+02 1.8E+05

ADH5;ADH5P4 5.0E+01 1.9E+05 3.6E+05 4.5E+04 2.4E+05 5.0E+01

CA2 1.2E+06 3.4E+06 7.0E+06 2.9E+06 2.5E+06 8.8E+05

IPI00101961 7.3E+05 1.2E+05 2.4E+05 1.8E+05 2.2E+05 1.9E+05

IGLV2-18 5.9E+05 1.4E+05 7.5E+04 4.5E+04 3.3E+05 6.0E+04

APOH 4.9E+06 2.8E+07 7.3E+06 1.3E+06 8.4E+06 1.2E+07

LOC390956 5.6E+05 1.1E+02 1.6E+06 3.6E+05 2.5E+05 5.7E+05

IPI00003939 5.2E+05 5.6E+05 1.9E+05 5.0E+01 8.0E+04 6.2E+05

MYL12B 5.0E+01 5.0E+05 1.2E+06 7.6E+05 1.7E+05 5.0E+01

CFH 1.6E+07 2.3E+07 2.4E+07 1.2E+06 8.8E+06 2.4E+07

DECR1 4.0E+05 5.0E+01 2.9E+05 7.1E+04 2.2E+05 8.2E+04

LOC401847 1.8E+06 8.2E+05 4.0E+05 5.0E+01 3.4E+05 1.4E+06

PRSS7 5.9E+06 2.8E+06 8.2E+06 5.0E+06 1.6E+06 2.6E+06

NUDT21 2.6E+05 1.0E+05 2.8E+05 2.1E+05 1.4E+05 1.9E+04

LOC652128 1.0E+07 4.3E+06 3.2E+06 3.5E+02 3.6E+06 6.6E+06

IGLC1;IGLV1-

44;IGLV1-40;IGLV3-

21;IGLV2-11;IGLV2-

14;IGL@;IGLC2;IGLC

3

3.6E+07 1.7E+07 2.0E+07 1.1E+06 1.4E+07 2.7E+07

A2M 9.7E+07 1.0E+08 1.1E+08 8.6E+06 6.1E+07 1.1E+08

TFF3 2.0E+06 1.4E+06 4.8E+06 2.0E+06 1.9E+06 8.6E+05

ENO1 4.9E+06 3.9E+06 4.9E+06 1.5E+06 2.8E+06 3.8E+06

TCN1 1.5E+05 3.0E+05 6.8E+05 4.0E+04 1.1E+05 5.1E+05

IGLC1;IGLV1-

44;IGLV1-40;IGLV3-

21;IGLV2-11;IGLV2-

14;IGL@;IGLC2;IGLC

3

5.0E+01 5.0E+01 9.6E+05 2.0E+02 1.2E+03 5.8E+05

C20orf114 2.0E+06 1.5E+05 3.8E+05 7.4E+05 1.9E+05 6.0E+05

IPI00382478 5.0E+06 2.6E+06 1.4E+06 1.4E+02 1.7E+06 3.8E+06

IGHG1;IGHG2;IGHV4

-31;IGH@

5.0E+01 5.5E+05 1.6E+06 5.0E+01 1.3E+06 5.0E+01

IPI00760721 9.6E+05 6.4E+05 1.0E+05 3.0E+04 4.4E+05 5.8E+05

MUC2 2.2E+07 4.1E+06 1.2E+07 4.3E+06 1.1E+07 8.6E+06

FH 8.4E+05 4.8E+05 1.2E+06 7.9E+05 6.3E+05 1.3E+05

AZGP1 7.0E+06 1.2E+07 4.9E+06 4.1E+06 4.3E+06 6.6E+06

HDGF 3.7E+06 1.9E+05 1.9E+05 5.0E+01 5.0E+01 2.6E+06

SELENBP1 6.2E+04 6.5E+05 4.8E+05 5.0E+01 1.0E+05 6.5E+05

IPI00383732 2.8E+06 2.5E+06 1.3E+06 6.2E+02 1.3E+06 3.0E+06

IGHA1 1.0E+06 8.0E+05 1.1E+06 5.0E+01 2.4E+05 1.6E+06


26

BCHE 5.1E+05 6.2E+05 1.0E+06 3.2E+04 5.3E+05 7.9E+05

DLD 6.5E+05 1.7E+05 5.3E+05 2.3E+05 5.4E+05 8.3E+04

IGLV8-61 7.5E+05 1.6E+05 2.0E+05 3.7E+02 1.9E+05 5.2E+05

WDR1 2.0E+06 1.4E+06 1.5E+06 9.4E+05 1.1E+06 1.1E+06

GALNT7 2.9E+05 1.4E+05 5.2E+05 4.2E+05 8.3E+04 1.1E+05

LAP3 9.7E+05 8.3E+05 6.2E+05 1.1E+05 7.6E+05 6.9E+05

HEBP1 2.0E+06 1.2E+06 1.4E+06 2.2E+05 2.1E+06 6.8E+05

HSPA5 6.1E+06 5.9E+06 8.2E+06 6.4E+06 4.1E+06 2.7E+06

NME1;NME2 1.0E+06 5.0E+01 6.3E+05 1.1E+05 4.4E+05 5.4E+05

ADH6 1.4E+05 5.9E+05 1.3E+06 1.7E+05 9.7E+05 2.2E+05

C4BPA 5.2E+06 6.9E+06 4.7E+06 4.0E+05 3.3E+06 7.5E+06

IPI00550731 5.3E+07 5.7E+07 6.6E+07 2.7E+07 3.4E+07 5.6E+07

HBZ 2.0E+06 1.3E+06 1.9E+06 5.0E+01 6.3E+05 2.9E+06

IPI00830122 1.2E+05 9.6E+04 5.0E+01 5.0E+01 4.0E+04 1.0E+05

CNDP2 9.8E+05 1.4E+06 1.8E+06 1.8E+06 5.4E+05 5.5E+05

ACADVL 1.2E+06 8.4E+05 2.3E+05 2.3E+05 1.2E+06 1.2E+05

IGKV4-1 4.0E+06 4.3E+06 6.1E+06 1.5E+06 2.4E+06 5.9E+06

TF 8.8E+07 1.4E+08 8.4E+07 1.2E+07 8.1E+07 1.2E+08

IGHG4 1.0E+07 2.5E+07 1.6E+07 8.1E+05 5.7E+06 2.8E+07

HPR 7.9E+06 1.4E+07 9.0E+06 8.8E+05 5.3E+06 1.5E+07

IPI00382486 3.3E+05 8.6E+04 6.2E+04 5.0E+01 7.7E+04 2.5E+05

ACE 8.3E+05 1.4E+05 7.1E+05 1.5E+05 3.4E+05 6.9E+05

IPI00007899 4.3E+06 8.7E+05 4.5E+05 1.5E+05 1.2E+06 2.5E+06

MYL6B 1.9E+06 6.8E+05 1.2E+06 1.6E+05 1.5E+06 1.0E+06

SERPINA1 4.1E+07 8.3E+07 5.0E+07 1.6E+07 3.6E+07 7.3E+07

AKR1A1 3.6E+05 7.9E+05 8.1E+05 6.1E+05 7.5E+05 5.8E+04

TTR 1.5E+07 1.6E+07 5.1E+06 2.0E+06 6.3E+06 1.8E+07

ALB 5.9E+08 7.8E+08 5.3E+08 1.0E+08 4.0E+08 8.9E+08

HPR;HP 1.5E+07 2.2E+07 1.1E+07 1.3E+06 9.8E+06 2.4E+07

CALR 4.1E+06 2.7E+06 4.5E+06 4.7E+06 2.7E+06 9.8E+05

PGM2 4.3E+05 2.5E+05 4.4E+05 3.5E+05 3.1E+05 1.7E+05

UGT2B15 2.3E+05 2.7E+05 1.5E+06 1.4E+06 3.0E+02 1.1E+05

IPI00029863 1.9E+06 4.0E+06 1.7E+06 1.7E+06 2.5E+06 1.5E+06

HBB 9.4E+06 1.8E+07 3.1E+07 4.0E+05 2.0E+07 2.4E+07

IPI00003470 1.8E+06 1.9E+05 2.1E+05 7.5E+02 1.6E+05 1.5E+06

ADA 1.1E+07 7.1E+06 5.5E+06 3.8E+06 7.6E+06 6.7E+06

CLCA1 3.9E+07 1.5E+07 4.1E+07 3.8E+06 4.5E+07 2.5E+07

SLPI 1.1E+07 1.8E+05 8.1E+06 1.4E+07 2.9E+05 7.2E+05

IGHG1;IGHG2;IGHV4

-31;IGH@

3.6E+05 3.5E+05 2.0E+05 1.4E+05 2.0E+05 3.6E+05

SERPINA6 1.3E+06 2.4E+06 6.6E+05 5.0E+01 1.2E+06 2.2E+06

CS 3.0E+05 3.1E+02 5.0E+01 1.2E+05 5.0E+01 1.1E+05

GOT2 2.3E+05 1.9E+05 3.8E+05 1.9E+05 1.9E+05 2.5E+05

HSPB1 2.3E+05 5.0E+01 6.5E+05 1.2E+05 4.8E+05 8.2E+04

KRT6B 7.7E+05 3.2E+03 2.2E+06 1.2E+06 4.0E+05 7.2E+05


27

IPI00384397 7.1E+06 1.5E+06 1.4E+06 7.3E+04 2.3E+06 5.6E+06

LOC651751 5.0E+01 3.4E+05 3.6E+05 3.6E+05 2.0E+05 5.0E+01

HPX 2.7E+07 4.1E+07 1.2E+07 1.6E+06 2.0E+07 4.2E+07

IPI00386131 2.7E+05 8.5E+04 5.0E+01 8.2E+02 5.0E+01 2.9E+05

IGHG1;IGHG2;IGHV4

-31;IGH@

5.1E+07 3.1E+07 3.8E+07 4.2E+06 2.3E+07 7.2E+07

PEBP1 6.2E+06 6.3E+06 6.9E+06 1.8E+06 2.4E+06 1.2E+07

PCK1 3.6E+05 6.2E+05 1.0E+06 4.5E+05 8.8E+05 3.5E+05

IPI00383887 1.7E+05 9.3E+04 5.0E+01 5.0E+01 5.8E+04 1.6E+05

GSR 8.2E+05 5.3E+05 8.5E+05 5.5E+05 7.5E+05 5.3E+05

ITLN1 3.3E+06 2.5E+06 1.2E+06 4.4E+05 3.0E+06 2.4E+06

C3 6.1E+07 8.0E+07 8.8E+07 9.4E+06 6.8E+07 1.2E+08

LOC653879 2.1E+07 2.7E+07 2.3E+07 5.3E+06 1.9E+07 3.8E+07

IPI00890703 3.0E+05 1.2E+05 2.8E+05 2.1E+02 5.9E+04 5.4E+05

MTTP 6.5E+06 2.1E+06 3.6E+06 3.1E+06 5.9E+06 1.6E+06

AMBP 1.0E+06 2.9E+06 2.8E+06 7.7E+04 3.1E+06 2.7E+06

IPI00382499 1.1E+06 8.5E+05 2.4E+06 1.6E+06 6.3E+05 1.5E+06

LTF 2.3E+06 3.0E+06 3.3E+06 3.8E+06 3.3E+06 5.9E+05

CFHR3 7.3E+05 5.3E+05 1.2E+05 1.8E+05 2.6E+05 8.0E+05

PEPD 2.3E+06 2.7E+05 6.3E+05 1.8E+06 5.0E+01 1.1E+06

UBA1 5.0E+01 5.0E+01 7.0E+04 5.0E+01 9.2E+01 6.3E+04

IPI00829980 7.3E+05 2.7E+05 4.5E+05 5.0E+01 2.3E+05 1.1E+06

KRT14 4.5E+05 2.4E+05 7.8E+05 7.2E+05 4.1E+05 2.3E+05

ITGB1 9.0E+05 2.7E+05 1.7E+05 4.7E+05 5.0E+05 2.7E+05

IPI00007906 6.2E+05 5.8E+05 2.2E+05 1.1E+05 3.9E+05 8.1E+05

GOT1 2.6E+05 2.8E+05 8.5E+04 6.8E+04 3.1E+05 2.0E+05

SERPINA5 2.0E+06 1.3E+06 1.1E+06 2.0E+05 1.3E+06 2.6E+06

LDHA 6.5E+05 3.5E+05 5.4E+05 1.1E+02 7.9E+05 6.6E+05

IPI00384404 1.2E+05 8.4E+04 2.6E+05 5.0E+01 1.1E+05 3.4E+05

IPI00384391 2.3E+06 4.4E+05 5.7E+05 4.2E+04 8.1E+05 2.4E+06

GNB2 5.7E+05 5.0E+01 9.6E+04 1.5E+05 3.2E+05 1.9E+05

PZP 1.1E+07 1.3E+07 1.1E+07 5.0E+06 9.4E+06 2.1E+07

CKMT1B;LOC10013

3623;CKMT1A

6.5E+05 2.4E+05 4.4E+05 6.3E+05 5.6E+05 1.4E+05

C6orf58 3.5E+06 5.9E+05 4.3E+06 2.1E+06 3.4E+06 3.0E+06

GNB1 5.0E+01 1.2E+05 2.6E+05 3.3E+05 5.5E+04 5.0E+01

IGHG1;IGHG2;IGHV4

-31;IGH@

1.4E+05 5.0E+01 5.0E+01 4.1E+02 5.0E+01 1.4E+05

LOC652113 2.1E+05 5.0E+01 1.2E+06 9.8E+05 3.9E+05 8.9E+04

ANXA5 7.5E+05 5.0E+01 1.6E+05 2.0E+05 3.0E+05 4.4E+05

HBD 1.1E+07 1.4E+07 2.9E+07 4.0E+05 1.9E+07 3.6E+07

EFHD2 1.9E+05 5.0E+01 1.5E+05 2.6E+05 1.6E+04 8.2E+04

ZG16B 2.1E+06 2.4E+06 4.5E+06 5.4E+06 1.6E+06 2.5E+06

IGKV1D-8 2.5E+06 1.7E+06 1.3E+06 3.5E+05 1.6E+06 3.8E+06

UGT2B10 6.9E+05 2.1E+05 6.6E+05 1.5E+06 5.0E+01 1.6E+05


28

FGB 6.2E+07 2.6E+07 1.6E+07 6.8E+06 4.9E+07 5.6E+07

ANXA4 5.4E+05 7.0E+05 1.3E+06 1.8E+06 5.6E+05 3.5E+05

GSTP1 2.6E+06 2.6E+06 6.4E+06 9.5E+06 1.7E+06 1.3E+06

ALDOC 5.2E+05 5.2E+05 9.7E+05 1.7E+06 2.8E+05 2.0E+05

FLNB 4.0E+06 1.4E+06 3.0E+06 2.7E+06 3.7E+06 2.9E+06

MASP1 3.5E+05 2.1E+05 1.1E+05 3.0E+04 1.5E+02 7.1E+05

UGCGL1 2.3E+05 1.2E+05 7.2E+04 5.0E+01 1.0E+05 3.5E+05

CNDP2 6.3E+05 7.0E+05 1.1E+06 1.2E+06 4.7E+05 1.0E+06

DEFA1;LOC728358 3.2E+06 4.2E+06 2.2E+06 1.0E+07 1.3E+05 2.0E+05

IGHG3 8.3E+07 7.6E+07 4.8E+07 1.5E+07 5.2E+07 1.6E+08

IGLV3-12 2.3E+05 5.0E+01 5.0E+01 5.0E+01 5.0E+01 2.6E+05

SCARB2 4.6E+05 2.0E+05 8.2E+05 1.1E+06 5.1E+05 1.2E+05

TPI1;RCTPI1 6.8E+05 3.3E+05 4.9E+05 1.6E+05 8.9E+05 6.6E+05

OGDH 3.5E+05 7.1E+05 1.1E+06 1.4E+06 9.4E+05 1.4E+05

C4A 1.9E+07 4.4E+07 2.8E+07 3.4E+06 4.5E+07 5.9E+07

UGT1A1;UGT1A8 4.5E+05 5.5E+05 5.0E+05 6.6E+05 3.6E+05 7.5E+05

PIP 5.0E+01 2.8E+05 8.6E+05 1.2E+06 1.4E+05 5.0E+01

VDAC2 3.0E+05 8.7E+04 6.2E+05 6.1E+05 5.9E+05 5.0E+01

CBR3 1.9E+06 1.9E+06 1.5E+06 4.5E+06 1.2E+06 6.6E+05

MUC5AC 4.1E+07 6.1E+06 5.7E+07 1.0E+08 1.5E+07 7.3E+06

C1QB 1.8E+05 1.6E+05 2.3E+04 3.6E+02 5.0E+01 4.4E+05

UGT2A3 9.1E+05 2.0E+05 9.1E+04 8.1E+05 3.7E+05 2.9E+05

IPI00743194 1.9E+06 3.7E+05 4.1E+05 3.5E+05 6.2E+05 2.4E+06

MAOB 3.1E+05 6.0E+04 3.1E+05 1.9E+05 3.3E+05 3.3E+05

RBP4 4.5E+06 5.9E+06 2.6E+06 2.5E+05 6.1E+06 1.0E+07

CA1 1.1E+06 2.7E+06 4.2E+06 4.8E+05 5.3E+06 4.4E+06

PSMA6 4.6E+05 1.6E+05 1.6E+06 2.2E+05 1.8E+06 7.8E+05

ANO10 5.0E+01 1.1E+05 1.5E+05 1.4E+05 1.9E+05 5.0E+01

HLA-DRB3;HLA-

DRB5;LOC649783;L

OC100133484;LOC1

00133661;HLA-

DRB4;LOC10013381

1;HLA-DRB1

5.0E+01 1.4E+05 8.9E+04 9.1E+04 2.1E+05 5.0E+01

IPI00384407 1.8E+06 4.5E+05 5.5E+05 2.3E+02 8.8E+05 2.7E+06

FKBP1A 1.3E+05 1.7E+05 3.0E+05 4.2E+04 2.2E+05 5.3E+05

ENPP7 1.9E+05 5.0E+01 5.0E+01 4.9E+04 1.7E+05 3.1E+04

ACADS 6.8E+05 3.5E+05 6.2E+05 4.9E+05 1.5E+06 1.8E+05

GKN2 5.0E+01 5.0E+01 1.8E+05 9.1E+04 1.5E+05 5.0E+01

GPI 2.0E+06 1.1E+06 1.1E+06 2.9E+06 1.7E+06 9.9E+05

DDT 6.1E+05 3.9E+05 1.1E+06 1.1E+06 7.6E+05 9.6E+05

IGHG1;IGHG2;IGHV4

-31;IGH@

5.4E+05 6.8E+04 1.1E+05 3.1E+02 1.4E+05 8.4E+05

ESD 9.7E+04 5.0E+01 1.7E+05 6.9E+04 1.5E+05 1.6E+05

TKT 1.5E+06 7.3E+05 1.8E+06 1.8E+06 2.6E+06 1.5E+06


29

IGHG1;IGHG2;IGHV4

-31;IGH@

1.1E+07 1.7E+07 7.9E+06 4.2E+06 1.7E+07 3.2E+07

IGLC1;IGLV1-

44;IGLV1-40;IGLV3-

21;IGLV2-11;IGLV2-

14;IGL@;IGLC2;IGLC

3

1.7E+05 5.0E+01 5.0E+01 5.0E+01 3.7E+04 2.1E+05

TGM3 3.8E+06 2.7E+05 1.3E+06 7.8E+06 1.4E+05 1.4E+05

KRT10 4.9E+06 4.0E+06 1.2E+06 1.0E+06 8.1E+06 6.4E+06

IPI00385252 1.1E+05 5.0E+01 5.0E+01 4.8E+02 5.0E+01 1.7E+05

NDUFAB1 5.5E+04 5.0E+01 1.5E+05 1.8E+05 1.6E+05 5.0E+01

IPI00829827 1.8E+06 2.0E+05 2.2E+05 5.1E+04 8.7E+05 2.6E+06

KRT5 2.4E+06 6.7E+05 1.5E+06 4.7E+06 1.5E+06 1.2E+06

ADH4 1.3E+06 1.0E+06 2.1E+06 1.7E+06 4.7E+06 8.9E+05

IGKV2-40 1.5E+06 4.6E+05 6.7E+05 8.3E+05 9.8E+05 2.5E+06

HSD17B4 1.2E+06 4.0E+05 9.2E+05 2.3E+06 1.2E+06 6.3E+05

CAT 2.0E+06 1.2E+06 1.2E+06 1.3E+06 3.0E+06 3.1E+06

ADH1A 6.7E+06 4.7E+06 9.5E+06 4.1E+06 2.0E+07 1.0E+07

FGL1 8.8E+04 4.5E+05 1.1E+05 5.9E+04 6.3E+05 4.0E+05

CMBL 2.2E+06 1.5E+06 2.4E+06 4.3E+06 3.9E+06 2.0E+06

PSCA 1.1E+05 4.6E+03 1.4E+06 2.0E+06 4.1E+04 5.5E+05

CKMT2 2.0E+05 3.2E+05 1.1E+06 4.4E+05 2.1E+06 2.5E+05

IGHG1;IGHG2;IGHV4

-31;IGH@

2.0E+05 1.8E+05 1.6E+05 1.9E+05 2.6E+05 4.9E+05

REG1A 9.7E+06 3.5E+06 1.0E+07 1.1E+07 2.1E+07 8.7E+06

AKR1C2 1.9E+06 3.3E+06 4.5E+06 1.3E+07 2.3E+06 2.0E+06

LOC652694 3.5E+06 1.5E+06 1.2E+06 1.0E+06 3.4E+06 6.4E+06

KRT16 1.1E+06 8.4E+04 4.8E+04 1.9E+06 1.9E+05 1.4E+05

LOC100133944 6.8E+05 8.5E+05 1.9E+06 3.9E+05 4.6E+06 1.1E+06

LOC652493 9.4E+04 5.0E+01 1.2E+05 1.8E+02 6.5E+04 3.2E+05

CTSD 1.2E+06 5.7E+05 1.3E+06 3.1E+06 1.8E+06 6.4E+05

FCGBP 3.4E+07 1.6E+07 2.4E+07 3.4E+07 6.6E+07 3.4E+07

HBA1;HBA2 2.8E+07 2.3E+07 4.5E+07 2.5E+06 5.6E+07 1.1E+08

GDI1 2.6E+05 1.7E+05 6.7E+05 2.6E+05 3.8E+05 1.4E+06

ADH1C 4.0E+05 8.0E+05 1.4E+06 2.6E+06 1.7E+06 4.1E+05

CD163L1 6.8E+05 5.0E+01 3.2E+03 6.7E+05 5.0E+05 8.0E+04

AKR1B10 5.8E+06 3.4E+06 8.2E+06 1.3E+07 1.3E+07 7.0E+06

LTA4H 4.4E+05 1.5E+05 5.0E+05 1.1E+06 5.3E+05 4.3E+05

GAPDH 7.9E+06 4.1E+06 1.1E+07 2.2E+07 1.3E+07 9.6E+06

CFHR1 1.8E+05 3.9E+05 7.0E+05 5.7E+04 6.9E+05 1.7E+06

IPI00914985 1.7E+05 5.0E+01 5.0E+01 5.0E+01 7.8E+04 2.6E+05

LCN2 2.6E+06 2.0E+05 4.1E+05 6.3E+06 2.1E+05 6.5E+04

PGRMC2 2.2E+05 5.0E+01 5.0E+01 2.9E+05 1.6E+05 5.0E+01

PGLS 5.6E+05 1.4E+05 7.8E+04 7.1E+05 6.4E+05 2.6E+05

SYCN 8.0E+05 1.5E+05 3.8E+03 1.5E+06 2.6E+05 1.8E+05


30

DNASE1 1.8E+06 2.5E+04 5.0E+01 2.2E+06 7.3E+05 9.0E+05

IPI00784773 2.4E+06 3.9E+05 4.1E+05 1.4E+05 2.5E+06 4.2E+06

ACADM 2.9E+05 1.0E+05 1.8E+05 4.8E+04 9.8E+05 2.3E+05

LGALS3 1.2E+06 7.9E+05 1.7E+06 4.7E+06 3.0E+06 5.5E+05

MSRA 1.1E+05 5.0E+01 5.0E+01 8.9E+04 1.0E+05 6.5E+04

IGHD 2.4E+05 7.4E+04 2.4E+05 5.0E+01 9.9E+05 2.9E+05

TFF2 3.3E+06 2.5E+05 8.7E+05 8.5E+06 1.0E+06 6.9E+05

HPR 3.3E+06 1.0E+06 6.9E+06 2.6E+06 1.2E+07 1.2E+07

NAPRT1 3.8E+05 9.1E+04 1.7E+05 1.0E+06 2.7E+05 3.3E+05

RNPEP 9.5E+05 4.1E+05 8.4E+05 4.2E+06 1.2E+06 4.9E+05

IPI00387024 2.4E+06 3.5E+05 1.2E+05 9.4E+04 3.5E+06 4.1E+06

GAPDHS 2.8E+05 5.0E+01 1.7E+05 7.8E+04 6.6E+05 5.0E+05

SERPINB6 1.6E+05 5.0E+01 1.1E+05 3.0E+05 3.1E+05 1.5E+05

ZG16 2.3E+06 1.7E+05 8.7E+05 1.4E+06 5.0E+06 3.1E+06

VSIG1 5.0E+01 5.0E+01 8.0E+05 2.3E+06 9.1E+02 5.0E+01

TPI1;RCTPI1 3.0E+06 5.0E+06 1.6E+06 1.2E+07 1.2E+07 4.3E+06

NPM1 1.1E+05 2.6E+05 1.4E+05 4.8E+05 9.3E+05 6.7E+04

AKR1C1 5.7E+05 7.4E+04 4.0E+05 1.7E+06 6.9E+05 8.6E+05

IPI00387109 5.0E+01 5.0E+01 4.2E+04 5.0E+01 1.3E+05 5.0E+01

EPHX1 6.2E+05 4.6E+05 9.8E+05 5.5E+06 7.7E+05 2.6E+05

ANXA1 2.2E+05 2.8E+05 6.6E+05 3.6E+06 4.7E+04 1.4E+05

FN1 2.3E+05 5.0E+01 3.9E+02 1.3E+03 3.6E+04 6.9E+05

C1RL 1.5E+05 3.7E+05 3.9E+04 1.9E+05 9.8E+05 6.5E+05

KRT2 2.9E+06 1.2E+06 5.8E+05 1.4E+06 8.3E+06 5.8E+06

LGALS4 3.6E+06 9.8E+05 1.6E+06 1.2E+07 7.1E+06 3.0E+06

LOC100130811 5.3E+04 5.0E+01 5.0E+01 5.4E+02 5.0E+01 2.0E+05

C1QC 3.1E+05 1.8E+05 1.4E+05 6.4E+04 4.3E+05 1.9E+06

MUC5B 6.5E+06 2.3E+06 4.5E+06 4.9E+07 1.4E+06 7.3E+05

FTH1 1.3E+05 5.0E+01 5.0E+01 5.0E+01 3.1E+05 1.8E+05

KRT71 5.9E+05 5.0E+01 5.0E+01 5.0E+01 1.3E+06 1.0E+06

IPI00384395 8.1E+04 5.0E+01 5.0E+01 5.0E+01 1.8E+05 1.6E+05

REG1B 5.7E+04 4.7E+04 2.0E+05 3.5E+05 7.3E+05 2.4E+05

GP2 8.6E+05 2.2E+04 5.0E+01 1.9E+05 1.8E+06 1.9E+06

RBKS 5.0E+01 5.0E+01 6.7E+04 1.7E+05 6.5E+04 5.6E+04

CPA4 8.9E+05 1.2E+05 7.1E+04 2.1E+06 1.3E+06 1.5E+06

IGHV1OR15-1 2.8E+06 1.4E+06 3.1E+05 2.5E+04 9.1E+06 1.1E+07

SERPINB1 1.7E+06 9.7E+05 2.0E+06 1.8E+07 2.7E+06 2.8E+06

IPI00383629 7.7E+05 1.5E+05 5.0E+01 9.2E+04 1.5E+06 3.0E+06

PRSS3 1.9E+06 3.3E+05 1.2E+05 7.1E+06 1.7E+06 2.8E+06

KRT1 4.0E+06 2.8E+06 2.1E+06 7.1E+06 2.3E+07 1.5E+07

APCS 5.2E+05 3.5E+05 2.9E+05 1.2E+05 1.5E+06 4.3E+06

MUC1 1.4E+05 1.1E+02 4.7E+04 7.7E+05 1.7E+05 4.4E+04

IPI00829752 1.3E+05 5.0E+01 5.0E+01 5.0E+01 2.2E+05 4.4E+05

IPI00748265 7.8E+04 5.0E+01 4.7E+04 4.7E+02 1.1E+05 5.8E+05

TAGLN2 1.1E+06 1.1E+05 2.5E+05 6.9E+06 6.8E+05 9.7E+05


31

CUZD1 2.6E+06 4.7E+06 1.3E+06 1.6E+07 2.4E+07 9.6E+06

CLPS 2.9E+05 3.7E+05 2.2E+05 1.6E+06 6.1E+05 3.0E+06

PNLIPRP2 1.5E+07 5.9E+06 8.2E+05 4.3E+07 6.1E+07 3.6E+07

IL1RN 8.5E+04 1.2E+03 5.0E+01 4.6E+05 5.0E+01 9.2E+04

KRT6A 7.5E+04 1.3E+03 5.0E+01 4.9E+05 5.0E+01 5.0E+01

KLK1 9.5E+05 6.9E+05 6.8E+05 9.1E+06 5.8E+06 5.0E+05

IPI00382442 4.5E+05 5.0E+01 5.0E+01 1.6E+05 7.9E+05 2.2E+06

KRT9 2.5E+06 4.1E+05 1.5E+05 3.8E+06 8.5E+06 8.9E+06

HBE1 8.5E+05 3.6E+05 3.1E+05 2.2E+04 5.9E+06 5.1E+06

IGK@ 6.8E+04 5.0E+01 5.0E+01 5.0E+01 1.2E+05 4.0E+05

CELA2B 9.5E+05 8.5E+05 5.6E+05 1.4E+07 7.6E+06 1.3E+06

CST2 5.0E+01 1.1E+05 5.0E+01 1.1E+06 5.7E+03 5.0E+01

HEXA 6.7E+04 5.0E+01 5.0E+01 4.2E+05 1.8E+05 8.9E+04

IPI00827637 7.2E+04 5.0E+01 5.0E+01 5.0E+01 3.6E+05 4.4E+05

RNASE1 6.9E+05 1.3E+06 2.5E+06 2.9E+07 2.0E+07 2.9E+06

CPA2 7.9E+06 4.2E+06 3.8E+06 1.6E+08 2.4E+07 1.4E+07

PSMB1 1.6E+05 5.0E+01 5.0E+01 9.1E+05 7.7E+05 3.3E+05

S100A7 4.3E+04 1.4E+05 1.6E+05 4.8E+06 5.5E+04 1.1E+05

LY6D 5.0E+01 6.4E+04 5.0E+01 9.1E+05 4.0E+04 5.0E+01

CEL 1.4E+07 5.8E+06 3.6E+06 1.1E+08 2.1E+08 3.1E+07

CTRB1 1.0E+07 2.7E+06 1.5E+06 4.9E+07 1.4E+08 3.2E+07

CTRL 4.1E+05 4.2E+05 1.3E+05 6.7E+06 6.1E+06 1.9E+06

PRSS1 9.4E+06 4.2E+06 3.5E+06 1.5E+08 9.1E+07 3.3E+07

PSMA1 1.3E+05 5.0E+01 4.4E+04 1.5E+06 9.9E+05 4.6E+05

CTRL 2.0E+05 5.0E+01 5.0E+01 4.6E+05 2.3E+06 6.5E+05

CPA1 1.8E+07 4.5E+06 5.6E+06 2.9E+08 1.5E+08 4.3E+07

IPI00385254 7.4E+04 5.0E+01 5.0E+01 3.7E+04 8.4E+05 4.4E+05

KRT76 2.1E+05 5.0E+01 5.9E+04 1.1E+06 2.5E+06 1.2E+06

NME1 2.2E+05 5.0E+01 5.0E+01 2.6E+06 8.3E+05 5.8E+05

CELA3B 1.6E+06 2.4E+04 7.8E+05 3.1E+07 1.1E+07 4.4E+06

PNLIPRP1 3.1E+06 3.0E+05 4.7E+05 4.9E+07 2.2E+07 6.7E+06

FASN 1.5E+05 7.6E+04 5.5E+04 2.8E+04 1.7E+06 4.7E+06

CELA2A 2.3E+06 9.2E+05 3.9E+05 4.3E+07 3.3E+07 1.3E+07

IPI00219910 2.3E+05 8.1E+04 1.4E+05 5.1E+06 5.7E+06 6.1E+05

IPI00827829 6.4E+04 5.7E+04 3.2E+02 3.3E+06 5.0E+01 5.0E+01

PNLIP 2.8E+07 3.5E+06 7.6E+06 7.0E+08 2.7E+08 9.1E+07

PRSS2 2.6E+06 2.7E+05 6.5E+05 5.3E+07 3.6E+07 1.1E+07

CLC 5.0E+01 4.3E+04 5.0E+01 7.0E+05 3.7E+05 2.1E+05

CELA3A 4.4E+06 1.1E+06 2.2E+06 1.3E+08 7.7E+07 2.7E+07

CTRC 2.0E+06 1.2E+06 4.9E+05 4.4E+07 5.6E+07 1.2E+07

MPO 5.0E+01 7.1E+04 3.5E+04 3.4E+06 2.2E+05 5.0E+01

GP2 8.7E+04 2.3E+05 1.1E+05 1.0E+07 2.6E+06 2.2E+06

GP2 9.5E+05 8.4E+04 4.8E+05 1.4E+07 2.8E+07 1.4E+07

CPB1 4.0E+06 1.5E+06 9.4E+05 1.3E+08 8.8E+07 2.4E+07

AMY1C;AMY2A;AMY 2.1E+06 3.3E+05 5.1E+05 7.7E+07 3.3E+07 4.6E+06


32

1A;AMY1B

AMY2B 1.0E+07 4.9E+06 3.9E+06 4.3E+08 3.2E+08 4.6E+07

SERPINI2 5.0E+01 6.7E+04 5.0E+01 3.6E+06 3.3E+06 2.9E+05

PLA2G1B 3.7E+05 4.9E+04 7.6E+04 4.0E+07 1.5E+07 3.8E+06

HMGCL 5.0E+01 5.0E+01 5.0E+01 5.0E+01 4.1E+04 5.0E+01

CD302;LY75 5.0E+01 2.7E+02 5.0E+01 1.1E+05 5.0E+01 5.0E+01

HPN 5.0E+01 8.8E+02 5.0E+01 3.4E+05 5.0E+01 5.0E+01

SCFV 5.0E+01 5.0E+01 5.0E+01 1.5E+02 5.0E+01 5.4E+04

IPI00884092 5.0E+01 5.0E+01 5.0E+01 5.0E+01 3.0E+04 3.5E+04

AZU1 5.0E+01 2.3E+03 5.0E+01 1.4E+06 3.4E+05 5.0E+01

TPM3 5.0E+01 5.0E+01 5.0E+01 5.0E+01 3.5E+04 7.6E+04

LOC100132635 5.0E+01 4.2E+03 5.0E+01 3.1E+06 6.2E+04 5.0E+01

ERO1L 5.0E+01 5.0E+01 5.0E+01 9.8E+04 6.0E+04 5.0E+01

IPI00827580 5.0E+01 5.0E+01 5.0E+01 5.0E+01 4.5E+04 1.2E+05

TTN 5.0E+01 5.0E+01 5.0E+01 4.4E+04 4.1E+04 8.1E+04

PGAM4;LOC728188 5.0E+01 5.0E+01 5.0E+01 1.3E+05 6.4E+04 5.0E+01

IPI00398220 5.0E+01 5.0E+01 5.0E+01 5.2E+02 5.0E+01 1.9E+05

BPI 5.0E+01 1.4E+03 5.0E+01 2.9E+06 5.0E+01 5.0E+01

CTRB2 5.0E+01 5.0E+01 5.0E+01 2.7E+05 1.2E+05 5.0E+01

C1QA 5.0E+01 5.0E+01 5.0E+01 3.8E+05 8.5E+04 5.0E+01

GLO1 5.0E+01 9.9E+01 5.0E+01 4.5E+05 2.2E+05 5.0E+01

AMY1C;AMY2A;AMY

1A;AMY1B

5.0E+01 9.4E+02 5.0E+01 2.7E+06 1.3E+06 5.0E+01

GALM 5.0E+01 5.0E+01 5.0E+01 3.0E+05 2.9E+05 5.0E+01

GC 5.0E+01 5.0E+01 5.0E+01 5.0E+01 6.7E+05 5.0E+01

CNP 5.0E+01 5.0E+01 5.0E+01 7.9E+05 5.8E+04 5.0E+01

LCN1 5.0E+01 1.2E+02 5.0E+01 1.4E+06 5.0E+01 5.0E+01

MBL2 5.0E+01 5.0E+01 5.0E+01 1.1E+06 2.6E+04 5.0E+01

CDH1 5.0E+01 5.0E+01 5.0E+01 7.4E+05 4.2E+05 4.2E+04

IGLC1;IGLV1-

44;IGLV1-40;IGLV3-

21;IGLV2-11;IGLV2-

14;IGL@;IGLC2;IGLC

3

5.0E+01 5.0E+01 5.0E+01 5.0E+01 9.6E+05 6.5E+05

PRSS1 5.0E+01 5.0E+01 5.0E+01 2.9E+06 3.0E+05 4.2E+05

GFPT1 5.0E+01 5.0E+01 5.0E+01 4.6E+06 7.8E+05 1.0E+05

Suppl Table 3:

Map PMID


33

A1 15542607

A2 11274345

A3 11606045

A4 12205041

B1 10791986

B2 16611744

B3 16122376

B4 10880430

B5 11950845

B6 16614236

B7 11368003

C1 17915332

C2 10339425

C3 12049736

C4 9164868

C5 11389839

C6 11505407

C7 Gerhard Michal: Biochemical Pathways: An Atlas of Biochemistry and

Molecular Biology

D1 15071501

D2 15716280

D3 9054461

D4 7925482

D5 Gerhard Michal: Biochemical Pathways: An Atlas of Biochemistry and


34

Molecular Biology

E1 12235123


carboxyl-ester lipase maturity-onset diabetes of the young...

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