key: cord-0322535-id5k4370 authors: Mansell, T.; Barwon Infant Study Investigator Group,; Saffery, R.; Burugupalli, S.; Ponsonby, A.-L.; Tang, M. L.; O'Hely, M.; Bekkering, S.; Smith, A. A. T.; Rowland, R.; Ranganathan, S.; Sly, P. D.; Vuillermin, P.; Collier, F.; Meikle, P.; Burgner, D. title: Early life infection and proinflammatory, atherogenic metabolomic and lipidomic profiles at 12 months of age: a population-based cohort study date: 2021-12-02 journal: nan DOI: 10.1101/2021.12.02.21267173 sha: 07be0b96f87fe6cc0e66e94b2a6442a02632779b doc_id: 322535 cord_uid: id5k4370 Background: The risk of adult onset cardiovascular and metabolic (cardiometabolic) disease accrues from early life. Infection is ubiquitous in infancy and induces inflammation, a key cardiometabolic risk factor, but the relationship between infection, inflammation, and metabolic profiles in early childhood remains unexplored. We investigated relationships between infection and plasma metabolomic and lipidomic profiles at age 12 months, and mediation of theseassociations by inflammation.Methods: Matched infection, metabolomics and lipidomics data were generated from 555 infants in a pre-birth longitudinal cohort. Infection data from birth to 12 months were parent-reported (total infections at age 1, 3, 6, 9, and 12 months), inflammation markers (high-sensitivity C-reactive protein, hsCRP); glycoprotein acetyls GlycA) were quantified at 12 months. Metabolic profiles were 12-month plasma nuclear magnetic resonance metabolomics (228 metabolites) and liquid-chromatography/mass-spectrometry lipidomics (776 lipids). Associations were evaluated with multivariable linear regression models. Results: Frequent infant infections were associated with adverse metabolomic (elevated inflammation markers, triglycerides, phenylalanine, and lower HDL cholesterol, apolipoprotein A1, and omega-3 fatty acids) and lipidomic profiles (elevated phosphatidylethanolamines and lower hexosylceramides, trihexosylceramides, and cholesteryl esters). Similar, more marked, profiles were observed with higher GlycA, but not hsCRP. GlycA, but not hsCRP, mediated a substantial proportion of the relationship between infection and metabolome/lipidome. Conclusions: Infants with a greater infection burden from birth to 12 months had pro-inflammatory and pro-atherogenic plasma metabolomic/lipid profiles, indicative of heightened risk of cardiovascular disease, obesity, and type 2 diabetes in adults. These findings suggest potentially modifiable pathways linking early life infection and inflammation with subsequent cardiometabolic risk.Funding: The establishment work and infrastructure for the BIS was provided by the Murdoch Childrens Research Institute, Deakin University and Barwon Health. Subsequent funding was secured from the National Health and Medical Research Council of Australia, The Jack Brockhoff Foundation, the Scobie Trust, the Shane OBrien Memorial Asthma Foundation, the Our Womens Our Childrens Fund- Raising Committee Barwon Health, The Shepherd Foundation, the Rotary Club of Geelong, the Ilhan Food Allergy Foundation, GMHBA Limited and the Percy Baxter Charitable Trust, Perpetual Trustees. In-kind support was provided by the Cotton On Foundation and CreativeForce. Research at Murdoch Childrens Research Institute is supported by the Victorian Government's Operational Infrastructure Support Program. This work was also supported by NHMRC Senior Research Fellowships (1008396 to ALP; 1064629 to DB; 1045161 to RS) and NHMRC Investigator Grants to ALP (1110200) and DB (1175744). SB is supported by the Dutch Research Council (452173113). (elevated inflammation markers, triglycerides, phenylalanine, and lower HDL 48 cholesterol, apolipoprotein A1, and omega-3 fatty acids) and lipidomic profiles 49 (elevated phosphatidylethanolamines and lower hexosylceramides, 50 trihexosylceramides, and cholesteryl esters). Similar, more marked, profiles were 51 observed with higher GlycA, but not hsCRP. GlycA, but not hsCRP, mediated a 52 . CC-BY 4.0 International license It is made available under a perpetuity. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. ; https://doi.org/10.1101/2021. 12 is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. Infectious diseases are ubiquitous in infancy and childhood, with potential long-term 81 impacts on health across the life course. Infection has been recognised as a 82 potential contributor to atherosclerotic cardiovascular disease (CVD), one of the 83 leading causes of adult morbidity and mortality, since the 19 th century (Nieto, 1998) . is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. ; https://doi.org/10.1101/2021. 12 . 02.21267173 doi: medRxiv preprint metabolic perturbations in later childhood and adulthood (Feingold & is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint At the 4-week, 3-month, 6-month, 9-month and 12-month time points following birth, 137 mothers were asked to report each episode of infant illness or infection since the 138 previous timepoint using standardised on-line questionnaires. The number of parent-139 reported infections from birth to 12 months was defined as the total number of 140 respiratory tract infections, gastroenteritis, conjunctivitis and acute otitis media 141 episodes from birth to the 12-month timepoint. It was not possible to identify the 142 proportion of parent-reported infections that lead to health service 143 utilisation (Rowland et al., 2020) . is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint Breastfeeding duration up to 12 months of age was collected from maternal 153 questionnaire data. As most evidence for the protective effect of breastfeeding on 154 early life infection is from comparisons between any breastfeeding and no 155 breastfeeding (Victora et al., 2016) , and in light of previous evidence in BIS for an 156 association between even a short duration of breastfeeding and lower odds of 157 infection in early infancy (Rowland et al., 2020) , we first looked at breastfeeding as a 158 binary (any/none) measure in models (presented in the main text). As most infants 159 (98.2%) were breastfed to some extent, and it is unknown the degree to which 160 breastfeeding, and the timing of breastfeeding, might affect 12-month metabolomics 161 and lipidomics, we also considered duration of breastfeeding to 12 months of age for 162 sensitivity analyses (see Supplementary Files 1C is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. To complement GlycA as a measure of inflammation, high sensitivity C-reactive 206 protein (hsCRP) was also quantified in 12-month plasma using ELISA assay (R&D 207 is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. ; https://doi.org/10.1101/2021.12.02.21267173 doi: medRxiv preprint The flowchart for the 555 infants included in this study is shown in Figure 1 , and the 268 cohort characteristics for these infants are shown in Table 1 . The median number of 269 total parent-reported infections from birth to 12 months of age was 5 (IQR = [3 to 5]). is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. ; https://doi.org/10.1101/2021.12.02.21267173 doi: medRxiv preprint There was evidence for higher number of infections associating with higher 284 inflammatory markers (GlycA and hsCRP), lower HDL, HDL2, and HDL3 285 cholesterols, smaller HDL particle size, lower ApoA1, docosahexaenoic acid (DHA), 286 lower total omega-3 fatty acids, lower citrate, higher phenylalanine, and to a lesser 287 extent with higher triglycerides and lower sphingomyelins (Figure 2a, Supplementary 288 File 1C). In models with GlycA as the marker of infection burden, metabolomic 289 differences observed for higher GlycA were largely similar to, but more marked than, 290 those for parent-reported infections; including cholesterols (lower HDL, higher LDL 291 and very-large-density lipoprotein (VLDL) cholesterol), apolipoproteins (lower ApoA1, 292 higher apolipoprotein B (ApoB)), higher total fatty acids, higher total triglycerides and 293 cholines, amino acids (higher phenylalanine, isoleucine, and glycine, lower histidine), is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint Higher hsCRP was associated with lower HDL cholesterol, ApoA1 and histidine, and 305 higher phenylalanine, as observed for GlycA, and with lower levels of most other 306 amino acids, total fatty acids, total cholesterols, albumin, phosphoglycerides and 307 VLDL triglycerides, and higher LDL triglycerides (Figure 3a, Supplementary File 1E) . is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. Higher hsCRP was also associated with differences across several lipid classes, 345 particularly lower plasmalogens, lysophosphatidylcholines, and phosphatidylcholine 346 classes (Figure 5a, Supplementary File 1I) . 347 There was stronger correlation for lipidomic differences related to infection and 348 GlycA (r=0.77) than for infection and hsCRP (r=0.33) (Figure 4c, Figure 5b) . is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. We next assessed whether inflammation (i.e., GlycA or hsCRP at 12 months) 361 mediated the effects of infection on specific metabolite and lipid measures (adjusted 362 p-value < 0.1) ( is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint Measures shown in Table 2 are the metabolomic and lipidomic measures that were associated with number of parent-reported infection in 377 linear regression models, as shown in Figure 2 and Figure 4 . Models are adjusted for infant age at sample collection, infant sex, birth weight z-score, maternal household income during pregnancy, 379 maternal smoking during pregnancy, breastfeeding (any/none), and sample time from collection to storage. In this study, cumulative parent-reported infection burden from birth to 12 months 382 was associated with adverse NMR metabolomic and LC/MS lipidomic profiles at 12 383 months of age. Similar but more marked effects on these profiles were evident when 384 considering GlycA, a cumulative inflammation marker, as a surrogate of infection 385 burden. In contrast, differences in metabolomic and lipidomic profiles associated with 386 higher hsCRP were largely distinct from, and less marked than, those for GlycA, 387 suggesting that GlycA may be superior to hsCRP as an early life marker of infection 388 burden. There was evidence that inflammation (GlycA, but not hsCRP) may partly 389 mediate many of the largest metabolomic and lipidomic differences. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. ; https://doi.org/10.1101/2021.12.02.21267173 doi: medRxiv preprint reactant that increases rapidly following acute stimulus and returns to baseline levels 431 within a matter of days and is therefore mostly used as a diagnostic adjunct in 432 children with acute infection or inflammation (Gabay & Kushner, 1999 is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. With the approved ethics for this study, the individual participant data cannot be 503 made freely available online. Interested parties can access the data used in this 504 study upon reasonable request, with approval by the Barwon Infant Study data 505 custodians. As part of this process, researchers will be required to submit a project 506 concept for approval, to ensure the data is being used responsibly, ethically, and for 507 scientifically sound projects. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprint The copyright holder for this this version posted December 2, 2021. ; https://doi.org/10.1101/2021.12.02.21267173 doi: medRxiv preprint SARS-CoV-2 infection dysregulates 609 the metabolomic and lipidomic profiles of serum Do childhood infections 611 contribute to adult cardiometabolic diseases? The Pediatric infectious disease 612 journal Childhood hospitalisation with infection 615 and cardiovascular disease in early-mid adulthood: a longitudinal population-616 based study Early childhood hospitalisation with infection and 619 subclinical atherosclerosis in adulthood: the Cardiovascular Risk in Young 620 n-3 fatty acids, inflammation and immunity: new mechanisms to 622 explain old actions Increased arterial stiffness in HIV-625 infected children: risk factors and antiretroviral therapy Revised birth centiles for weight, 628 length and head circumference in the UK-WHO growth charts on 631 behalf of the Barwon Infant Study Investigator Comparison of the predictive value of GlycA and other biomarkers 659 of inflammation for total death, incident cardiovascular events, 660 noncardiovascular and noncancer inflammatory-related events, and total 661 cancer events Australian children aged 11-12 years and their parents The effect of inflammation and infection on 667 lipids and lipoproteins Inflammageing: chronic inflammation in ageing, 669 cardiovascular disease, and frailty Acute-phase proteins and other systemic responses 672 to inflammation Serum lipids in infection History of respiratory infections in the first 12 yr among children from a birth 678 cohort A novel protein glycan biomarker and LCAT activity in metabolic syndrome GlycA, a pro-inflammatory glycoprotein biomarker, and 684 incident cardiovascular disease: relationship with C-reactive protein and renal 685 function Improved LC/MS methods 687 for the analysis of metal-sensitive analytes using medronic acid as a mobile 688 phase additive Concordant peripheral lipidome signatures in two large 691 clinical studies of Alzheimer's disease Ala-693 Genome-wide study for circulating metabolites identifies 694 62 loci and reveals novel systemic effects of LPA Infection 697 and inflammation-induced proatherogenic changes of lipoproteins. The 698 Journal of infectious diseases Associations of GlycA and high-sensitivity C-701 reactive protein with measures of lipolysis in adults with obesity U-shaped relationship of HDL and risk of infectious disease: two 709 prospective population-based cohort studies Plasma lipid profiling shows similar associations with 713 prediabetes and type 2 diabetes GlycA, a novel marker for low grade inflammation, reflects gut 716 microbiome diversity and is more accurate than high sensitive CRP in 717 reflecting metabolomic profile GlycA and hsCRP are independent and 720 additive predictors of future cardiovascular events among patients undergoing 721 angiography: the intermountain heart collaborative study Early atherosclerosis in humans: 724 role of diffuse intimal thickening and extracellular matrix proteoglycans Infections and atherosclerosis: new clues from an old 727 hypothesis? Physiology of leptin: energy homeostasis, 729 neuroendocrine function and metabolism Altered 731 serum lipid profile after systemic infection in children: risk factor for CHD? 732 Antiinflammatory therapy with 735 canakinumab for atherosclerotic disease The biomarker GlycA is associated with chronic 739 inflammation and predicts long-term risk of severe infection Burden of infection in Australian infants Cutting Edge: Severe SARS-CoV-2 Infection in Humans Is Defined 746 by a Shift in the Serum Lipidome Inflammation, infection and atherosclerosis. Trends in 749 cardiovascular medicine Quantitative serum nuclear magnetic resonance metabolomics in 752 cardiovascular epidemiology and genetics R: A language and environment for statistical computing Mediation: R 757 package for causal mediation analysis Estimates of the global, regional, and national morbidity, mortality, 760 and aetiologies of lower respiratory infections in 195 countries, 1990-2016: a 761 systematic analysis for the Global Burden of Disease Study Age-related prevalence of common upper 765 respiratory pathogens, based on the application of the FilmArray Respiratory 766 panel in a tertiary hospital in Greece Breastfeeding in the 21st century: epidemiology, 769 mechanisms, and lifelong effect Cohort Profile: The Barwon Infant Study Cytomegalovirus infection and relative risk of cardiovascular disease 775 (ischemic heart disease, stroke, and cardiovascular death): a meta-analysis of 776 prospective studies up to 2016 Plasma lipid profiles discriminate bacterial from viral 780 infection in febrile children Plasma lipid profiling in a large population-based cohort Metabolite profiling and cardiovascular event risk: a 786 prospective study of 3 population-based cohorts Quantitative serum nuclear magnetic resonance 789 metabolomics in large-scale epidemiology: a primer on-omic technologies High-throughput quantification of circulating 794 metabolites improves prediction of subclinical atherosclerosis Regression models for count data in 797 The authors thank the BIS participants for the generous contribution they have made 491 is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprintThe copyright holder for this this version posted December 2, 2021. is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprintThe copyright holder for this this version posted December 2, 2021. ; https://doi.org/10.1101/2021.12.02.21267173 doi: medRxiv preprint It is made available under a perpetuity.is the author/funder, who has granted medRxiv a license to display the preprint in (which was not certified by peer review) preprintThe copyright holder for this this version posted December 2, 2021. ; https://doi.org/10.1101/2021.12.02.21267173 doi: medRxiv preprint