City, USA). CA, Redwood QIAGEN (IPA®, Analysis Pathway Ingenuity® QIAGEN’s using analyzed the using controlled was (FDR) rate the of model linear a using performed were groups studied between T-testsmoderated expression raw Differential on [6]. applied intensities was RMA USA). CA, Clara, (Santa microarrays 2.0 ST Human (TableLPL) (wild-type 1). RNAblood Whole on hybridized were samples ( mutations LPL null for heterozygotes CM 20 ( LPLactivity of 5% than less and LPLmutations null for homozygotes LPLD 19 groups: three into divided were and study this in participated subjects 53 of total A to Methods was study this of aim The [5]. severe metabolism investigate profiles inacrossa CM spectrum ofLPL activity. of lipoproteins TG-rich cause of mendelian pathways LPLD rare with lipoproteins a individuals (TG)-rich is triglyceride and of (LPLD) hypertriglyceridemia deficiency key LPL a metabolism. is (LPL) lipase Lipoprotein Background LIPOPROTEINS CATABOLISM IN SEVERE HYPERTRIGLYCERIDEMIA CHYLOMICRONEMIA AND GENETIC FUNCTIONALAND INVESTIGATION OF LPL PATHWAYS INDEPENDENT OF TG-RICH Department of Medicine, Saguenay, of Medicine, Department QC, Canada Université Daniel Table 2: comparison of 2: Figure mediated clearance mechanisms (Figures2and 3). the receptor- of or systems docking pathways, signalling or circadian inflammatory,immune, Most 1). (Fig. by shared are 29 probes, differentially identified were ≥2-fold probes a gene and detected 5% in expressed of 142 FDR level, a significance <0.01, expression p-value change a at that revealed analyses expression than pancreatitis acute of prevalence higher HoLPL Results a =False discoveryFDR rate. HeLPL From HeLPL HoLPL Comparisons Affymetrix = Heterozygous LPL; Heterozygous = subjects presented significantly lower BMI and HDL-C mean values as well as well as values mean HDL-C and BMI lower significantly presented subjects vs vs Wild type LPL vs vs Wild type LPL

Differential expression analysis results Gaudet de Montréal Community Genomic Medicine Centre and ECOGENE-21 ECOGENE-21 and Centre Medicine Genomic Community Montréal de HoLPL Ethanol Degrada=on IV γ Mitochondrial L- LXR/RXR Ac=va=on IL-6 Oxida=ve Ethanol Degrada=on III P38 MAPK FaQy Acid Ac=va=on G- Coupled Receptor PPAR cAMP Role of Osteoblasts, Osteoclasts and Chondrocytes LPS/IL-1 Mediated Inhibi=on of RXR Func=on IL-10 Glycerol Degrada=on I Acetate Conversion to Acetyl-CoA Role of macrophages, Fibroblasts and Endothelial Cells α NAD Biosynthesis III PPARα/RXRα Ac=va=on Salvage Pathways of Pyrimidine Gα Inhibi=on of Matrix Fc Epsilon RI Oncosta=n IL-8 Glycogen Degrada=on III Agranulocyte Complement System Granulocyte Adhesion and IPA - ® Human Gene 2.0ST microarrays (ona total of48,226detected probes).

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Tremblay, PhD and IPAVennfrom of obtained diagram 1: Figure p NS: Bonferroni non-parametric using when significant distribution. the of a function Lipoprotein; High-Density HDL= Lipase; Lipoprotein LPL= stated. otherwise unless (SE), mean are Data controls. Consistently, this was associated with lower free glycerol and higher IL-1 higher plasma concentration, respectively (data not shown). and glycerol free lower with associated was this Consistently, controls. ( activity (IL)-1 LPL ( of limited absence total but in available CM and TG hyper severe 3 Figure Table 1: Geometric means and p-values obtained on log on obtained p-values and means Geometric HDL- Arterial Glucose, LDL- Total Total Ex or Alcohol Index (BMI), Body Mass Men:Women Age, Cardiovascular Diabetes Obesity Pancreatitis HeLPL HoLPL Common Clinical and and Clinical Translational Centre Research ≥ 0.1 HeLPL LPL years cholesterol cholesterol triglycerides β cholesterol current -adjusted Z-tests orExact Fischer Chi-Square. Significantly difference from n I- rcpos ee wr dw-euae in down-regulated were receptors IL-8 and , hypertension, abstinence, gene , n (%) mmol : n (%)

Characteristics ofstudyparticipants Differentially expressed biomarkers among biomarkers expressed Differentially mutations carrier , 48are specific to Reductionist representation of some IPA®some of representation Reductionist differentiatingpossibly pathways canonical , c n (%)

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CLEC4E CDA LINC00694 GAS5 DEFA1 CPA3 CHI3L1 C5AR1 ALPL ADGRG3 ACSL1 CSF2RB CSF2RB CPD CEACAM8 CASP5 BASP1 BACH1 ANXA3 Legend Identified pathways disease

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in in Chylomicron HoLPL -transformed data. -transformed HoLPL HoLPL ad in and ) MMP25 MMP25 MIR4802 MGAM2 MGAM MANSC1 LRG1 (CM) LOC100507006 HIST1H4L CYP4F3 CYP4F3 CXCR2 CXCR1 FPR2 FPR2 FFAR2 FCER1A FAM129A DOCK5 DOCK4 DDX11L2 4

Wild type LPL

® 24.9 (0.9) 52.8 (3.2) 4.9 (0.3) 1.3 (0.1) 2.6 (0.2) 4.5 (0.2) 1.1 (0.1) (n =14) 9 (64.3) 2 (14.3) 2 (14.3) and 7are specific to comparisons analyses: comparisons 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) APOC3 7:7 b normolipidemic

Post hoc Post

METTL12 METTL12 LOC102723340 RPL23A RNASE3 RBMX PI3 MS4A3 MS4A2 LOC729040 LINC00189 KCNJ15 IL1RAP IL1R2 IGKC HIST1H3H HCAR2 GZMK GK adipocytes adipocytes analyses performed using a C a using performed analyses LPL Pancreas Bonferroni 1

APOC3 Normal LPL activity or RXR RXR 30.0 (0.8) 14 (70.0) 0.8 (0.0) 1.4 (0.2) 7.2 (1.2) 8.7 (2.9) 56.6 (1.8) VLDL 8 (40.0) ? 6.1 (0.2) HoLPL (n =20) 7 (38.9) 7 (30.0) 4 (20.0) 9 (45.0) 4 (20.0) ? HeLPL α α FFA adjusted -LXR -FXR 13:7 otos Bod K n interleukins and GK Blood controls. Liver Liver ? ? ? HeLPL White blood cells gene expression Muscle cells cells Muscle IL1RAP/IL1R2 29 biomarkers are shared by shared are biomarkers 29

IL-1 JNK 1 ot hoc post HoLPL and 1 1 1 1 1 SIGLEC5 SIGLEC5

SNORD116-15 SNORD116-1 SNHG6 SNHG16 SNHG1 SERPING1

1

SLED1 SLED1 RPL27 REPS2 NOP56 NAMPT MME LUCAT1 LOC102724190 LOC101927851 LOC100507639

HeLPL HoLPL . Dunnett

2 HeLPL Defective LPL activity. LPL et.Al infcn pvle ( < .5 remain 0.05) < (p p-values significant All tests. 3 (Heterozygous) oprd to compared White blood cells gene expression 19 (100.0) significance level due to the to due level significance CXCR1/CR2 21.6 (2.6) 0.4 (0.0) 22.2 (1.0) , hn P atvt is activity LPL when ), 5.3 (0.2) 1.3 (0.1) 7.5 (0.9) 46.1 (3.2) 2 (10.5) 14 (73.7) NFkB IL-8 (n =19) 6 (31.6) 4 (21.1) 6 (31.6) 4 (21.0) ? HoLPL 2). Adaptation 2). ≥ (|FC| 7:12

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SNRPN SNRPN SNORD71 SNORD59A SNORD44 SNORD116-24

WDFY3 TRPM6 TECPR2 SNORD119 SNORD82 SNORD60 SNORD54 SNORD41 (Homozygous) HeLPL β and IL-8 and p-value <0.001 <0.001 <0.001 <0.001 <0.001 inegal 0.061 0.023 0.017 0.001 0.006 0.088 0.063 NS NS NS HoLPL

variance and loss

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neet o icoe Ti suy a spotd by supported was study This ECOGENE-21, a non-for-profit research organization. disclose. to Center of interest conflict Innovation no have Authors University Genome Canada). QC, (Montréal, the McGill at and performed Quebec were Microarrays staff. ECOGENE-21 and participants all to thankful are We Acknowledgements pancreatitis, functional investigations are ongoing. and (e.g.: epigenetic Genetic, approach. systems sub-phenotypes or cardiovascular CM mechanisms other LPLD of trajectory key risk clinical understand the differentiating further or reveal management. HoLPL lipoproteins between differences functional TG-rich and genetic Exploring of pathways independent LPL in involved mechanisms to key identify contribute may analyses functional with combined profiling, expression gene that highlight results These Conclusion [7] [6] Irizarry et al. 2003. [5] McGraw-Hill: 2789-2816; Molecular Basesof Inherited Disease [4] [3] [2] Maet al. 1991. [1] References CC1C2 ad L1 I12I1A) receptor (IL1R2/IL1RAP) genes. IL-1 and (CXCR1/CR2) IL-8 of down-regulation observed the with consistent to compared cytokines IL-1 pro-inflammatory of concentration Plasma in lower be to tended lipolysis of markers and glycerol free of concentration plasma and controls, and in down-regulated was HeLPL (e.g.: oxydation characteristics particles HDL and chylomicron, in differences significant revealed analyses Functional genes. expressed differentially involving pathways key in performed were analyses functional Complementary – Cont’dResults Ser Benjamini Gaudet Brunzell Monsalve Mattei β n I- wr sgiiaty ihr in higher significantly were IL-8 and

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and ih M Te lcrl iae G) pathway (GK) kinase glycerol The CM. with , 57:289-300. akr or markers et al. 1993. al. et et al. 2014. and [email protected] HeLPL et al. 1990. and Hochberg. 1995 Daniel HeLPL

Contact information:Contact Deeb cardiometabolic NEJM ih M a as cnrbt to contribute also may CM with

Gaudet . 2001. dt nt hw) wih is which shown), not (data Cytogenet NEJM Biostatistics ApoC HoLPL , 324:1671-1766; J

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