{"paper_id":"3c85b89b-ea97-4007-bb7c-244f4c658bed","body_text":"Inhibition of Endothelial Lipase by MEDI5884 Normalizes Phosphatidylinositol \nLevels in Coronary Artery Disease Patients  \nAnton I. Rosenbaum1*, Yue Huang1, Ruipeng Mu1, Kristina Kovacina1, ChaoYu \n(Denise) Jin2, B. Timothy Hummer3, Meina Liang1, Joseph S. Grimsby4** \nAffiliation:  \n1.  Integrated Bioanalysis,  \nClinical Pharmacology & Safety Sciences, R&D, AstraZeneca,  \n121 Oyster Point Boulevard, South San Francisco, CA 94080, USA \n2. Clinical Pharmacology and Quantitative Pharmacology \nClinical Pharmacology & Safety Sciences, R&D, AstraZeneca,  \n121 Oyster Point Boulevard, South San Francisco, CA 94080, USA \n3. CVRM Safety, Clinical Pharmacology and Safety Sciences, R&D, AstraZeneca, \nGaithersburg, MD 20878, USA \n4. Research and Early Development, Cardiovascular, Renal, and Metabolism, \nBioPharmaceuticals R&D, AstraZeneca, Gaithersburg, MD 20878, USA \n**Current address: Internal Medicine, Regeneron Pharmaceuticals, Inc., Tarrytown, NY \n10591, USA \ncorresponding author: \nanton.rosenbaum@astrazeneca.com \nT: +1-650-379-3099 \nOrcid: http://orcid.org/0000-0003-1939-951X\n \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \nAbstract: \nBackground. Endothelial lipase (EL) promotes high-density lipoproteins  (HDL) \nphospholipid degradation, increases catabolism of HDL and is an attractive target for \nthe potential treatment for cardiovascular disease. Inhibition of EL using a monoclonal \nneutralizing antibody, MEDI5884, demonstrated increased quantity and function of HDL. \nDeterminants of anti-atherosclerotic function of HDL comprise the interplay of various \ncomponents of HDL structure-activity relationship: size, shape and composition (lipid \nand protein). Previous studies have shown that single doses of MEDI5884 administered \nto healthy nonhuman primates (NHPs) and healthy subjects resulted in a dose-\ndependent increase in plasma phospholipids (PL) and that plasma PI levels in placebo \ntreated healthy subjects are significantly increased relative to CAD subjects participating \nin clinical trials NCT03001297 and NCT03351738, respectively.   \nMethods: Herein, we characterized using LC-MS/MS the plasma lipidome of NHPs, \nheathy subjects and subjects with coronary artery disease (CAD) following MEDI5884 \nadministration.  Results: MEDI5884 treated NHPs resulted in a prominent increase in \nphosphatidylinositols (PI) and cholesteryl esters (CE). Treatment with MEDI5884 \nrestores near-normal levels of PI in CAD patients. PI increases in both healthy subjects \nand CAD patients were dose-dependent, correlated with exposure and saturated at \napproximately 200 mg MEDI5884 subcutaneous (SC) dose in CAD patients. \nComparison of pharmacodynamic (PD) effects of repeat SC 200 mg doses of \nMEDI5884 in CAD patients revealed greater and more rapid increases in PI levels \ncompared to HDL-C and HDL phospholipid (HDL-PL). The increase in PI species was \ninversely correlated with decreases in free EL mass levels. Conclusions:  PI has \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \npreviously been shown to possess anti-atherosclerotic properties and led to increases in \nHDL cholesterol (HDL-C) and reverse cholesterol transport (RCT). The mechanism by \nwhich CE levels increase as the result of MEDI5884 administration can be attributed to \nthe observed increase in both substrates of the lecithin-cholesterol acyltransferase \n(LCAT) reaction: phosphatidylcholine/phosphatidylethanolamine (PC/PE) and \ncholesterol as the consequence of EL inhibition.  Further characterization of the \nunderlying biological mechanisms responsible for the decrease of the PI biomarker in \nCAD patient population relative to healthy subjects as well as in conjunction with \npharmacological intervention by MEDI5884 may reveal more information on this \nclinically-relevant biomarker and potential role in CAD.  \nIntroduction \nHDL removes cholesterol from macrophages and peripheral tissues and delivers \nit to the liver directly via scavenger receptor class B type 1 (SRB1) receptor or via low-\ndensity lipoproteins LDL and the LDL receptor for recycling or elimination via bile [1]. \nEndothelial lipase plays a critical role in maintaining the catabolism and homeostasis of \nHDL by hydrolyzing phospholipids and promoting the degradation of HDL particles and \nsubsequent elimination in urine. MEDI5884 is a neutralizing monoclonal antibody \nagainst endothelial lipase being studied for the treatment of coronary artery disease \n(CAD)[2, 3]. Since the failure of cholesteryl ester transfer protein (CETP) inhibitors to \nbring benefit to cardiovascular disease patients as it relates to augmenting HDL-C \nlevels one has to carefully consider understanding the functionality of HDL that goes \nbeyond cholesterol content ([4-7]). An interplay of structure activity relationship exists \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \nfor HDL where changes in its composition (proteome, lipidome) elicit changes in its \nstructure, shape, size and anti-atherosclerotic properties ([8]). \nPrevious studies have shown that single doses of MEDI5884 administered to \nhealthy monkeys resulted in a dose-dependent increase in HDL, plasma phospholipids \n(PL) and increased efflux. Further studies in HV and CAD patients revealed increases in \nHDL-C, HDL-PL and efflux [2, 9]. To elucidate the phospholipid changes resulting from \nthe administration of MEDI5884 we undertook a mass spectrometry-based lipidomic \nassessment which revealed a dose-dependent increase in phosphatidylinositols upon \nadministration of MEDI5884. \nResults \nEffects in Cynomolgus Monkeys \nLipidomic analyses revealed that PI species were substantially increased \ncompared to other phospholipids (Figure 1) in plasma from cynomolgus monkeys upon \nsingle dose administration of MEDI5884 (30 mg/kg).  Hierarchical clustering diagram \nrevealed that PI species cluster together and increase in their levels much more \ndrastically compared to other lipid species in a time-dependent fashion.  Furthermore, \nthis finding was confirmed using principle component analysis (PCA) which identified \nseveral sets of lipids clusters: PI and cholesterol esters (Figure 2).  \nCholesteryl esters are products of the lecithin–cholesterol acyltransferase (LCAT) \n(EC2.3.1.43) reaction where an sn2 acyl chain derived from phosphatidylcholine (PC) or \nphosphatidylethanolamine (PE) is transesterified to cholesterol [10]. The products of \nLCAT reaction are LysoPC and cholesteryl ester. Our lipidomic data revealed that both \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \nsubstrates of the LCAT reaction: PC/PE and cholesterol increased as the result of \nMEDI5884 treatment resulting in increased cholesteryl ester levels (Supplemental \nFigure 1). Possible explanations include activation of LCAT by a >30% increase in \nApoA1, enrichment of acidic phospholipid such as PI, increased recruitment of LCAT to \nHDL and/or increased synthesis of LCAT[11].  PI and CE lipids correlated well with \nconcomitant HDL-C increases as shown in Supplemental Figures 2 and 3, respectively. \nEffects in HV Subjects \nFollowing a single subcutaneous dose of MEDI5884 in study NCT03001297, \nincreases in the majority of plasma PI species were observed at all dose levels of \nMEDI5884 relative to the placebo (Supplemental Figure 4). The duration of the \nincreases appeared to correlate with MEDI5884 exposure in a dose-dependent manner; \nsubjects who received MEDI5884 600 mg showed the most sustained response post \nDay 28, which lasted until the end of the study (90 days) for some PI species. For the \nmajority of PI species, the maximal increase in plasma PI levels occurred on Day 21 \nand/or Day 28 and appeared to reach a plateau in the MEDI5884 300 mg dose group \nwhere there was a minimal difference in change compared with the MEDI5884 600 mg \ndose group consistent with maximum complete neutralization of circulating EL.  \nEffects in CAD Patients \nAs illustrated in Supplemental Figure 5 following 3 monthly SC doses of MEDI5884 \nin study NCT03351738, most plasma PI species dose-dependently increased relative to \nplacebo. The duration of increases in plasma PI appeared to correlate with MEDI5884 \nexposure. For most PI species, increases in PI levels reached saturation at the \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \nMEDI5884 350 mg dose level, and the MEDI5884 500 mg dose level did not result in \nfurther PI increases relative to baseline. However, on Day 91 at the MEDI5884 200 mg \ndose level, PI increases approached saturation levels observed in higher dose cohorts. \nPrevious studies have shown that PI lipids are substantially lower in CAD patients \ncompared to HV subjects[12]. We have endeavored to examine the effects of \nMEDI5884 on PI species in CAD patients relative to HV subjects. As is shown in \nFigure 3, a 200 mg SC monthly dose of MEDI5884 in CAD patients increased PI to \nlevels observed in healthy subjects thus normalizing their PI levels. \nEffects of MEDI5884 on Free Endothelial Lipase Levels in HV Subjects and CAD \nPatients \nFollowing administration of MEDI5884, dose-dependent suppression of EL levels was \nobserved (Supplemental Figures 6, 7 and 8). Complete suppression of EL levels was \nobserved for up to 45 days post dosing for the MEDI5884 300 and 600 mg dose groups \nfor HV. In CAD subjects, after 3 monthly SC MEDI5884 doses, dose-dependent \nsuppression of EL levelswas observed. EL levelsdecreased >85% from baseline \nthrough Day 91 at MEDI5884 350 and 500 mg doses. EL levels decreased >60% from \nbaseline through Day 91 at MEDI5884 200 mg. Initially, the 100 mg dose suppressed \nfree EL nearly completely, while the 50 mg dose achieved approximately 80% \nsuppression. Subsequently, EL suppression reversed consistently as MEDI5884 \nexposure decreased, in a dose-dependent manner.Greater and more rapid increases in \nPI levels compared to HDL-C and HDL-PL were observed, consistent with exposure to \nMEDI5884 [9] and with concomitant decreases in free EL mass levels (Figure 4). \nDiscussion \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \nEarly studies have shown that injection of PI into rabbits promotes cholesterol \nelimination and affects surface potential/charge of various lipoprotein particles, including \nHDL [13]. Further studies have shown that PI promotes cholesterol efflux from J774 \ncells and yet again injection of PI into rabbits promotes excretion of tritiated cholesterol \ninto bile indicating increased reversed cholesterol transport. Additionally, PI enriched \nHDL increases cholesterol update into HPG2 cells compared to PC-rich HDL [14]. \nPrevious studies have indicated that changes in lipid composition of HDL can affect its \nbioactivity [15]. PI along with other negatively charged phospholipids may impact the net \nsurface charge of HDL thereby modulating charge-dependent interactions with lipases, \nlipid transfer proteins, extracellular matrix, and other protein components (PMID: \n23543772). PI has been shown to alter the physical state of phosphatidylcholine \nsynthetic membranes and promote fluid phase formation and packing disorder [16]. \nAdditional studies have demonstrated an intermolecular interaction of \nphosphatidylinositol with the lipid raft molecules sphingomyelin and cholesterol [17]. \nThese data indicate that PI can affect cholesterol chemical potential thus promoting its \nefflux. Cholesterol chemical potential is an important variable in affecting its transport \nand trafficking [18]. Additionally, administration of PI has been shown to increase HDL-\nC levels in humans [19].  Furthermore, an association between low HDL-PI in subjects \nwith CAD status and high HDL-C have been reported previously [20]. Additionally, PI \nhas been shown to be decreased in acute-phase HDL (APHDL) obtained 34–38 h after \nsurgery from patients who underwent bypass surgical procedures [21, 22]. \nInterestingly, Stamler et al [13] have shown that PI inhibits esterification by LCAT and \nreduces CE levels. This differs from our observation in cynomolgus monkeys where we \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \nhave observed increased CE levels as the result of MEDI5884 administration and \nconcomitant PI level increases. This difference could be due to the fact we also \nobserved PC/PE level increases as the result of MEDI5884 administration. As \nmentioned above PC/PE are substrates for the LCAT reaction, and PI is not. One \npossible explanation is that in experiments conducted by Stamler et al. [13] reduction in \nPC as one of the substrates required for CE formation by LCAT due to administration of \nPI led to the reduced esterification rate.This could be especially true since LCAT activity \nwas measured using radioactive free cholesterol, not radioactive PC/PE. Additional \nexplanations include the potential species differences and pleiotropic effects of \nMEDI5884 administration. \nThe literature taken together indicates that PI is an important phospholipid that can \nmodulate lipoprotein particle charge, chemical potential of membranes and affect \ncholesterol transport in vivo promoting its elimination. Previous studies [2, 9] have \nshown that MEDI5884 treatment resulted in not only increased HDL-C quantity but also \nimproved quality as demonstrated by increased cholesterol efflux capacity. In \nconjunction with the finding that MEDI5884 promotes cholesterol efflux we can \nhypothesize that changes in plasma lipidome, in particular elevation of PI levels, as the \nresult of administration of MEDI5884 alters the structure activity relationship of HDL \nparticles to promote cholesterol efflux from macrophages.  \nIn conclusion, treatment with MEDI5884, an endothelial lipase neutralizing monoclonal \nantibody being developed for the treatment of coronary artery disease by increasing \nHDL quantity and function, restores near-normal levels PI in CAD patients on intensive \nstatin therapy in a dose-dependent manner. Further characterization of the underlying \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \nbiological mechanisms responsible for the decrease of the PI biomarker in CAD patient \npopulation relative to healthy subjects as well as in conjunction with pharmacological \nintervention by MEDI5884 may reveal more information on this clinically-relevant \nbiomarker and elucidate how changes in lipidome can affect cholesterol efflux capacity \nof HDL particles. Moreover, additional studies establishing that increasing PI in CAD \npatients impacts HDL function and clinical outcome would need to be conducted. \nMaterials and Methods \nGroups of 3 male cynomolgus macaques were dosed by single subcutaneous injection \nwith 0.5 mg/kg, 6 mg/kg, and 30 mg/kg of MEDI5884 (S6F1-4P), and 6 mg/kg of S1-\nIgG1. Plasma samples were collected 14 and 7 days prior to dosing as well as shortly \nprior to dosing and approximately 0.5, 1, 2, 3, 7, 14, 21, 28, 35, 42, 49, 56 days after \ndose. Further details on study design have been published previously[2].   \nTargeted Lipidomic Analysis for Monkey Plasma Samples \nAn automated electrospray ionization-tandem mass spectrometry approach was used, \nand data acquisition and analysis were carried out largely as described previously \n(Devaiah et al., 2006; Bartz et al., 2007) with some modifications as described in \nsupplemental information. \nPI quantification for clinical analyses was performed as described previously (). \nQuantification results of the levels of various PI species in healthy volunteers’ samples \nwere based on a total of 214 samples from study NCT03001297 [2] and a total of 978 \nplasma samples from Coronary Artery Disease (CAD) subjects participating in clinical \ntrial NCT03351738 [9].  \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \nFree Endothelial Lipase Assay \nSandwich immunoassay has been developed for the detection human EL utilizing ECL \nreadout.  A MSD plate (MesoScale Discovery) was coated with 5 µg/mL of MEDI5884, \nat 50 µl/well, incubated at 4°C on a flat surface and subsequently washed 3x with 300 \nµl/well of ELISA Wash Buffer. The assay plate(s) were blocked with 150 µl/well of \nI-Block Buffer (IBB) for \n≥ 60min on an orbital plate shaker at room temperature (RT). \nReference standards (RS), quality control (QC) and negative control (NC), prepared in \nIBB, and test samples were added to the plates at 35 µl/well. Samples were tested \nusing previously determined minimally required dilution (MRD, 2 for plasma samples). \nThe assay plate(s) were incubated at RT on a plate shaker with gentle shaking for 60 \nminutes ± 10 minutes. Unbound analyte was removed by washing the plate(s) with \nELISA wash buffer. To detect the captured analyte, 1 µg/ml of biotinylated anti-huEL \nmonoclonal antibody (OriGene) was added at 35 µl/well and incubated for additional 60 \nminutes ± 10 minutes at RT on a plate shaker. Unbound detection antibody was \nremoved by washing the plate(s). Streptavidin Sulfo-TAG was added at 35 µl/well and \nincubated for additional 60 minutes ± 10 minutes at RT on a plate shaker with gentle \nshaking. Plate(s) were washed again and Read Buffer added at 150 µl/well. Signal was \nread on a MSD sector imager instrument within 20 minutes. The ECL values for each \nplate were collected using the MSD Sector Imager. The ECL values for the reference \nstandard curves for each plate were plotted with Softmax Pro GxP v6.4 software \n(Molecular Devices, Sunnyvale, CA) using a 1/y\n2-weighted 4-Parameter Logistic (4-PL) \nmodel of curve fitting. The concentrations of unknown samples were interpolated from \nthe respective standard curves.  The Softmax-derived data was then imported into \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \nMicrosoft Excel Software and Spotfire (TIBCO® Spotfire® Analyst 7.9.2 HF-011 Build \nversion 7.9.2.0.12) to generate data reports and graphs. \nHDL-C and HDL-PL were analyzed as described previously [2]\n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \nFigures \nFigure 1. Hierarchical clustering analysis of cyno plasma phospholipidomic dataset after a single SC dose of MEDI5884 \n(30 mg/kg). \n \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \n \n \nFigure 2. Principal component analysis (PCA) of phospholipidomic data from cynomolgus monkey plasma upon \nadminstration of MEDI5884 at 0.5, 6, and 30 mg/kg (all timpoints) identifies several lipid clusters: PI and CE lipids. \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \n \nFigure 3. 200 mg SC monthly dose of MEDI5884 in CAD patients increased PI to levels observed in healthy subjects. \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \n \nFigure 4. Comparison of pharmacodynamic effects of 3 repeat monthly SC 200 mg doses of MEDI5884 (denoted with \narrows) in CAD patients. Pharmacodynamic biomarkers were HDL-C, HDL-PL, PI (18:0/18:1) and free EL mass, which \nwere evaluated after the first and third MEDI5884 administrations. \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \nAcknowledgment: The authors are or were employees of AstraZeneca at the time this work was conducted and may \nhold stock ownership and/or stock options or interests in the company. This study was funded by AstraZeneca. \nSupplemental Figures \n  \nSupplemental Figure 1. A. LCAT Reaction. B. Increase in CE as well as LCAT substrates Cholesterol, PE and PC after \nsingle SC dose of MEDI5884 (30 mg/kg) in cynomolgus monkeys. \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \nSupplemental Figure 2. Correlation analysis of normalized PI Levels vs. normalized HDL-C Levels from cynomolgus \nmonkeys upon administration of MEDI5884 (all dose levels and all timepoints). Analysis was performed in Spotfire. \n  \n \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \n \nSupplemental Figure 3. Correlation analysis of normalized cholesteryl esters levels vs. normalized HDL-C levels from \ncynomolgus monkeys upon administration of MEDI5884 (all dose levels and all timepoints). Analysis was performed in \nSpotfire. \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \n \nSupplemental Figure 4. Plasma PI Area % Change from Baseline Over Time (US Population, As Treated Population) from\nthe NCT03001297 study in HV following a single SC dose of MEDI5884. Avg = average; SEM = standard error of the \nmean; US = United States. Trelis legend includes a number above the PI species name, which is the average area ratio \nfor each PI species for all timepoints for the placebo group. Error bars represent the SEM. \nm \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \n \n \nSupplemental Figure 5. Plasma PI Area Ratio % Change from Baseline over Time for All MEDI5884 Dose Levels and Placebo (As-treated \nPopulation) from the NCT03351738 study in CAD patients. Avg = average; StdErr = standard error of the mean. Trelis legend includes a number \nabove the PI species name, which is the average area ratio for each PI species at all time points for the placebo group. Error bars represent the \nStdErr. Day 1 was used as baseline. Screening visit was imputed as Day 0.MEDI5884 was administered as 3 monthly SC doses.\n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \n \n \nSupplemental Figure 6. Dose-dependent suppression of EL levels in HV (As Treated Population) participating in study \nNCT03001297 upon administration of a single SC dose of MEDI5884. Avg = average; error bars represent SEM = \nstandard error of the mean. \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \n \nSupplemental Figure 7. Dose-dependent suppression of EL levels (change from baseline) in CAD patients participating \nin study NCT03351738 upon administration of 3 monthly SC doses of MEDI5884. Avg = average; error bars represent \nSEM = standard error of the mean.MEDI5884 was administered as 3 monthly SC doses.  \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \n \nSupplemental Figure 8. Dose-dependent suppression of EL levels (ng/mL) in CAD patients participating in study \nNCT03351738 upon administration of 3 monthly SC doses of MEDI5884. Avg = average; error bars represent SEM = \nstandard error of the mean. \n \n.CC-BY 4.0 International licenseavailable under a \nwas not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint \n\n \n \n \n \n \n \n \n \nReferences \n1. Phillips, M.C., Molecular mechanisms of cellular cholesterol efflux. J Biol Chem, 2014. 289(35): p. 24020-9. \n2. Le Lay, J.E., et al., Blocking endothelial lipase with monoclonal antibody MEDI5884 durably increases high density \nlipoprotein in nonhuman primates and in a phase 1 trial. Sci Transl Med, 2021. 13(590). \n3. Ruff, C.T., et al., LEGACY: Phase 2a Trial to Evaluate the Safety, Pharmacokinetics, and Pharmacodynamic \nEffects of the Anti-EL (Endothelial Lipase) Antibody MEDI5884 in Patients With Stable Coronary Artery Disease. \nArterioscler Thromb Vasc Biol, 2021. 41(12): p. 3005-3014. \n4. Tall, A.R. and D.J. Rader, Trials and Tribulations of CETP Inhibitors.  Circ Res, 2018. 122(1): p. 106-112. \n5. Yamashita, S. and Y. 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It is made \nThe copyright holder for this preprint (whichthis version posted June 3, 2024. ; https://doi.org/10.1101/2024.05.30.596497doi: bioRxiv preprint","source_license":"CC-BY-4.0","license_restricted":false}