Caffeine Impairs Red Blood Cell Storage Quality by Dual Inhibition of ADORA2b Signaling and G6PD Activity

Caffeine Impairs Red Blood Cell Storage Quality by Dual Inhibition of ADORA2b Signaling and G6PD Activity | bioRxiv /* */ /* */ <!-- <!-- /*! * yepnope1.5.4 * (c) WTFPL, GPLv2 */ (function(a,b,c){function d(a){return"[object Function]"==o.call(a)}function e(a){return"string"==typeof a}function f(){}function g(a){return!a||"loaded"==a||"complete"==a||"uninitialized"==a}function h(){var a=p.shift();q=1,a?a.t?m(function(){("c"==a.t?B.injectCss:B.injectJs)(a.s,0,a.a,a.x,a.e,1)},0):(a(),h()):q=0}function i(a,c,d,e,f,i,j){function k(b){if(!o&&g(l.readyState)&&(u.r=o=1,!q&&h(),l.onload=l.onreadystatechange=null,b)){"img"!=a&&m(function(){t.removeChild(l)},50);for(var d in y[c])y[c].hasOwnProperty(d)&&y[c][d].onload()}}var j=j||B.errorTimeout,l=b.createElement(a),o=0,r=0,u={t:d,s:c,e:f,a:i,x:j};1===y[c]&&(r=1,y[c]=[]),"object"==a?l.data=c:(l.src=c,l.type=a),l.width=l.height="0",l.onerror=l.onload=l.onreadystatechange=function(){k.call(this,r)},p.splice(e,0,u),"img"!=a&&(r||2===y[c]?(t.insertBefore(l,s?null:n),m(k,j)):y[c].push(l))}function j(a,b,c,d,f){return q=0,b=b||"j",e(a)?i("c"==b?v:u,a,b,this.i++,c,d,f):(p.splice(this.i++,0,a),1==p.length&&h()),this}function k(){var a=B;return a.loader={load:j,i:0},a}var l=b.documentElement,m=a.setTimeout,n=b.getElementsByTagName("script")[0],o={}.toString,p=[],q=0,r="MozAppearance"in l.style,s=r&&!!b.createRange().compareNode,t=s?l:n.parentNode,l=a.opera&&"[object Opera]"==o.call(a.opera),l=!!b.attachEvent&&!l,u=r?"object":l?"script":"img",v=l?"script":u,w=Array.isArray||function(a){return"[object Array]"==o.call(a)},x=[],y={},z={timeout:function(a,b){return b.length&&(a.timeout=b[0]),a}},A,B;B=function(a){function b(a){var a=a.split("!"),b=x.length,c=a.pop(),d=a.length,c={url:c,origUrl:c,prefixes:a},e,f,g;for(f=0;f<d;f++)g=a[f].split("="),(e=z[g.shift()])&&(c=e(c,g));for(f=0;f<b;f++)c=x[f](c);return c}function g(a,e,f,g,h){var i=b(a),j=i.autoCallback;i.url.split(".").pop().split("?").shift(),i.bypass||(e&&(e=d(e)?e:e[a]||e[g]||e[a.split("/").pop().split("?")[0]]),i.instead?i.instead(a,e,f,g,h):(y[i.url]?i.noexec=!0:y[i.url]=1,f.load(i.url,i.forceCSS||!i.forceJS&&"css"==i.url.split(".").pop().split("?").shift()?"c":c,i.noexec,i.attrs,i.timeout),(d(e)||d(j))&&f.load(function(){k(),e&&e(i.origUrl,h,g),j&&j(i.origUrl,h,g),y[i.url]=2})))}function h(a,b){function c(a,c){if(a){if(e(a))c||(j=function(){var a=[].slice.call(arguments);k.apply(this,a),l()}),g(a,j,b,0,h);else if(Object(a)===a)for(n in m=function(){var b=0,c;for(c in a)a.hasOwnProperty(c)&&b++;return b}(),a)a.hasOwnProperty(n)&&(!c&&!--m&&(d(j)?j=function(){var a=[].slice.call(arguments);k.apply(this,a),l()}:j[n]=function(a){return function(){var b=[].slice.call(arguments);a&&a.apply(this,b),l()}}(k[n])),g(a[n],j,b,n,h))}else!c&&l()}var h=!!a.test,i=a.load||a.both,j=a.callback||f,k=j,l=a.complete||f,m,n;c(h?a.yep:a.nope,!!i),i&&c(i)}var i,j,l=this.yepnope.loader;if(e(a))g(a,0,l,0);else if(w(a))for(i=0;i (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0];var j=d.createElement(s);var dl=l!='dataLayer'?'&l='+l:'';j.src='//www.googletagmanager.com/gtm.js?id='+i+dl;j.type='text/javascript';j.async=true;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-M677548'); Skip to main content Home About Submit ALERTS / RSS Search for this keyword Advanced Search New Results Caffeine Impairs Red Blood Cell Storage Quality by Dual Inhibition of ADORA2b Signaling and G6PD Activity Monika Dzieciatkowska , Ariel Hay , Aaron Issaian , Gregory R. Keele , Shaun Bevers , View ORCID Profile Travis Nemkov , Julie A Reisz , Mark Maslanka , Daniel Stephenson , View ORCID Profile Amy L Moore , Xutao Deng , Mars Stone , Kirk C. Hansen , Steve Kleinman , View ORCID Profile Philip J Norris , Michael P. Busch , Grier P Page , Nareg Roubinian , Yang Xia , James C Zimring , View ORCID Profile Angelo D’Alessandro doi: https://doi.org/10.1101/2025.05.27.656446 Monika Dzieciatkowska 1 Department of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus , Aurora, CO, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Ariel Hay 2 University of Virginia , Charlottesville, VA, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Aaron Issaian 1 Department of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus , Aurora, CO, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Gregory R. Keele 3 RTI International, Research Triangle Park , NC, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Shaun Bevers 1 Department of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus , Aurora, CO, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Travis Nemkov 1 Department of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus , Aurora, CO, USA 4 Omix Technologies Inc , Aurora, CO, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for Travis Nemkov Julie A Reisz 1 Department of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus , Aurora, CO, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Mark Maslanka 1 Department of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus , Aurora, CO, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Daniel Stephenson 1 Department of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus , Aurora, CO, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Amy L Moore 3 RTI International, Research Triangle Park , NC, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for Amy L Moore Xutao Deng 5 Vitalant Research Institute , San Francisco CA, USA 6 Department of Laboratory Medicine, University of California San Francisco , CA, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Mars Stone Find this author on Google Scholar Find this author on PubMed Search for this author on this site Kirk C. Hansen 5 Vitalant Research Institute , San Francisco CA, USA 6 Department of Laboratory Medicine, University of California San Francisco , CA, USA 1 Department of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus , Aurora, CO, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Steve Kleinman 7 University of British Columbia , Victoria, British Columbia, Canada Find this author on Google Scholar Find this author on PubMed Search for this author on this site Philip J Norris 5 Vitalant Research Institute , San Francisco CA, USA 6 Department of Laboratory Medicine, University of California San Francisco , CA, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for Philip J Norris Michael P. Busch 5 Vitalant Research Institute , San Francisco CA, USA 6 Department of Laboratory Medicine, University of California San Francisco , CA, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Grier P Page 3 RTI International, Research Triangle Park , NC, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Nareg Roubinian 5 Vitalant Research Institute , San Francisco CA, USA 6 Department of Laboratory Medicine, University of California San Francisco , CA, USA 8 Kaiser Permanente Northern California Division of Research , Oakland, CA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Yang Xia 9 Southern Medical University of Changsha , China Find this author on Google Scholar Find this author on PubMed Search for this author on this site James C Zimring 2 University of Virginia , Charlottesville, VA, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Angelo D’Alessandro 1 Department of Biochemistry and Molecular Genetics, University of Colorado Denver – Anschutz Medical Campus , Aurora, CO, USA 4 Omix Technologies Inc , Aurora, CO, USA Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for Angelo D’Alessandro For correspondence: angelo.dalessandro{at}cuanschutz.edu Abstract Full Text Info/History Metrics Supplementary material Preview PDF Abstract Caffeine is the most widely consumed psychoactive substance globally, yet its peripheral physiological effects remain incompletely understood. Leveraging comprehensive data from 13,091 blood donors in the REDS RBC-Omics study, we identify caffeine as a significant modulator of red blood cell (RBC) storage quality and transfusion outcomes. Elevated caffeine levels were reproducible across multiple donations from 643 recalled donors, selected based on their extremes in hemolytic propensity. Both in the screening and recalled cohorts, higher caffeine levels were associated with disrupted RBC metabolism, characterized by reduced glycolysis, depletion of adenylate pools or 2,3-bisphosphoglycerate, and increased markers of oxidative stress and osmotic fragility, including kynurenine accumulation. These observations were recapitulated in plasma and RBCs of eight volunteers upon consumption of a cup of coffee independently of brewing method (Chemex vs espresso). Clinically, elevated caffeine correlated with increased hemolysis and lower post-transfusion hemoglobin increments, especially pronounced in recipients transfused with RBCs from donors carrying common polymorphisms in the ADORA2b gene, a key regulator of RBC metabolism in hypoxia. These human findings were mechanistically validated using a murine model deficient in ADORA2b, which demonstrated impaired glycolytic flux, compromised antioxidant defenses – including caffeine-dependent direct inhibition of recombinantly-expressed glucose 6-phosphate dehydrogenase, and decreased transfusion efficacy (lower hemoglobin increments, higher bilirubin post-transfusion), effects further exacerbated by caffeine exposure during storage. Our study positions caffeine consumption as a modifiable factor in blood transfusion practice, advocating for precision strategies that integrate genetic and exposome factors, and identifies metabolic interventions to enhance blood quality and clinical outcomes. One sentence summary Caffeine consumption and genetic variants in the ADORA2b receptor synergistically impair red blood cell metabolism and transfusion efficacy, revealing a modifiable exposome–gene interaction for precision transfusion medicine. INTRODUCTION Caffeine is one of the most widely consumed psychoactive compounds worldwide, primarily sourced from coffee, tea, energy drinks, and sodas ( 1 ). In the United States, over 67% of adults report daily coffee consumption, averaging more than 1.5 cups per day (∼135 mg), with 36% of adults reporting an average daily consumption of 3-5 cups. Globally, coffee consumption exceeds 10 billion kilograms annually and continues to grow at an estimated 5% per year ( 2 ). An 8-ounce cup of brewed coffee (∼240 ml) contains approximately 95 mg of caffeine, while a 12-ounce can of cola (∼355 ml) contains around 35 mg, and a standard energy drink may contain up to 160 mg. These widespread dietary exposures make caffeine one of the most pervasive modulators of systemic and cellular physiology. As a non-selective antagonist of adenosine receptors, caffeine – a purine alkaloid - exerts its primary effects through blockade of A1 and A2A receptors in the central nervous system, enhancing alertness and psychostimulation ( 3 ). However, peripheral adenosine receptors — notably the A2B subtype (ADORA2B) — are also affected. Mature red blood cells (RBCs), though anucleate, express functional ADORA2B ( 4 ), a G protein-coupled receptor that activates cAMP production and downstream kinases such as protein kinase A (PKA) and AMP-activated protein kinase (AMPK) ( 5 ). In hypoxic conditions, adenosine levels increase, activating ADORA2B to stimulate glycolysis and 2,3-bisphosphoglycerate (2,3-BPG) synthesis ( 4 ). This facilitates oxygen unloading at the tissue level ( 6 ) and supports redox balance by favoring fluxes through glycolysis at the expense of the pentose phosphate pathway (PPP) ( 4 ), thereby liming NADPH generation and thus NADPH-dependent glutathione recycling. The clinical relevance of these pathways becomes evident in the context of blood storage, when oxidant stress to RBCs is high ( 7 ). Transfusion of packed RBCs is the most common inpatient procedure after vaccination in the U.S., with over 12 million units transfused annually. RBCs are stored for up to 42 days at 1–6°C, during which they undergo progressive biochemical deterioration ( 7 ). This “storage lesion” includes depletion of ATP and 2,3-BPG, oxidative damage to proteins and lipids, vesiculation, and membrane loss ( 7 ). Importantly, the capacity of stored RBCs to regenerate 2,3-BPG and maintain ATP upon rewarming is tightly linked to their capacity to circulate in vivo after transfusion, a gold standard parameter that assesses the quality of transfused blood, also referred to as post-transfusion recovery (PTR) ( 8 ). Stored RBCs progressively lose the capacity to activate the PPP ( 9 ), resulting in decreased NADPH production and loss of antioxidant buffering capacity. This makes RBCs vulnerable to oxidative hemolysis ( 10 ), particularly in recipients with inflammatory conditions. Accumulation of purine catabolites such as hypoxanthine, a marker of oxidant stress and ATP degradation, further correlates with reduced RBC lifespan after transfusion ( 8 ). Therefore, pathways that preserve purine homeostasis and redox balance are critical targets for improving stored RBC quality. High-altitude physiology offers a natural model for understanding the role of purine metabolism in RBC energy and redox physiology ( 11 ). At elevations above 3,000 meters, plasma adenosine levels rise by up to 10-fold, activating ADORA2B in RBCs to increase 2,3-BPG and enhance tissue oxygen delivery ( 4 ). In humans and mice, this adaptation is blunted when adenosine is enzymatically degraded ( 12 ) or ADORA2B is genetically deleted. In murine models, pharmacologic activation of AMPK improves post-storage 2,3-BPG and ATP levels( 13 ). These mechanisms are beneficial to acclimatization to high altitude, to the extent that proteasomal degradation of the adenosine equilibrative nucleoside transporter ENT1 favors faster acclimatization upon reascent ( 14 ). Caffeine, by antagonizing ADORA2B, has the potential to impair this protective signaling. Additionally, caffeine has been reported to directly and competitively bind to G6PD— the rate-limiting enzyme of the PPP—inhibiting its activity and reducing NADPH production ( 15 ). Xu and colleagues report that, in vitro, caffeine - at concentrations as low as 50 μM (equivalent to 1–2 cups of coffee) - reduces G6PD activity by up to 40%. Thus, habitual caffeine consumption could plausibly affect the redox capacity and glycolytic balance of donor RBCs, particularly during storage. Beyond biological (age, BMI, sex) or genetic factors ( 16 ), the quality of stored blood is impacted by the so-called exposome, i.e., donors’ dietary, recreational, professional or medical exposures that are not grounds for blood donor deferral ( 17 ). Two otherwise healthy blood donors may produce RBC units of vastly different storage quality due to recent dietary intake, medications, or occupational exposures. Encouraged by interesting preliminary data ( 18 ), here we investigated the impact of caffeine on RBC storage quality, here we stratified 13,000 donors enrolled in the Recipient Epidemiology and Donor Evaluation Study (REDS) RBC Omics, with detailed genomic, metabolomic, and storage phenotype data. Leveraging this resource, we conducted a two-part investigation to determine how caffeine and ADORA2B signaling interact to shape RBC storage outcomes, including in vitro and post-transfusion hemolysis. Materials and Methods In the interest of space, extensive details for this section are provided in Supplementary Materials. REDS RBC Omics: Index and Recalled donors Metabolomics analyses were performed on day 42 packed RBCs from 13,091 “index” donors enrolled across four blood centers as part of the Recipient Epidemiology and Donor evaluation Study (REDS RBC Omics). Omics results were analyzed as a function of single nucleotide polymorphisms (SNPs) mapping on the region coding for ADORA2b, as extrapolated from genomics data on 879,000 single nucleotide polymorphisms (SNPs). Donors ranking in the 5 th and 95 th percentile for end-of-storage hemolysis (n=643) were invited to donate a second (“recalled”) unit, which was tested at storage days 10, 23 and 42 (1,929 samples) for hemolytic parameters, high-throughput metabolomics ( 19 – 21 ), proteomics ( 22 ), lipidomics ( 23 ). Results were analyzed as a function of caffeine levels and ADORA2b SNPs. Caffeine consumption in healthy volunteers To interrogate the metabolic effect of caffeine consumption we randomized eight volunteer to consume a coffee cup either brewed as chemex or espresso (n=4 per group). Blood samples were collected at baseline and after 45 min and 5h from consuming the beverage, and plasma and RBCs were separated via gentle centrifugation (10 min at 4°C at 2,000 g ) prior to metabolomics analyses. G6PD activity assay in presence of caffeine We recombinantly expressed and purified a human canonical G6PD or the deficient African variant (V68M; N126D – class III variant, 10-60% residual activity) – most prevalent in the REDS RBC Omics cohort (∼13% of donors of African descent and 2% of donors of Hispanic descent ( 24 )). Activity assays were performed as described ( 15 ), in presence or absence of 50 μM caffeine ( 15 ). Storage and Post-transfusion recovery (PTR) studies in ADORA2b mice Mouse post-transfusion recovery (PTR) studies were performed as previously described ( 25 ). Storage of ADORA2b KO mice (n=3) for 7 or 12 days was followed by transfusion into Ubi-GFP mice, which were used as recipients to allow visualization of the test cells in the non-fluorescent gate. To control for differences in transfusion and phlebotomy, mCherry (red fluorescent-labeled) RBCs were used as a tracer RBC population (never stored). These RBCs were added to stored RBCs immediately prior to transfusion. PTR was calculated by dividing the post-transfusion by the pre-transfusion ratio (Test/Tracer), with the maximum value set as 1 (or 100% PTR). In follow up experiments, PTR studies were repeated after supplementation of caffeine (100 μM – no. C0750 Sigma Aldrich) to citrate-phosphate-dextrose-adenine CPDA1 standard storage solutions ( 25 ). This concentration is consistent with physiological plasma caffeine levels in high-consumption donors (low to high range: 20–100 μM ( 18 )) and in vitro studies mimicking real-life exposures (range 50–100 μM ( 15 )). Determination of hemoglobin and bilirubin increment via the vein-to-vein database Association of ADORA2b alleles with hemoglobin increments was performed by interrogating the vein-to-vein database, as described in Roubinian et al.( 26 ). Analyses were performed on the whole REDS population and further stratified for caffeine measurements, with a focus on the upper 50 th percentile of caffeine measurements in REDS donors. RESULTS High levels of caffeine in donor red blood cells are reproducible across time To assess the extent and physiological impact of caffeine exposure on stored red blood cell (RBC) quality, we first quantified caffeine levels in packed RBC units from 13,091 donors enrolled in the REDS RBC Omics study. Caffeine levels were either below detection threshold or negligible in ∼5,000 donors, spanning a broad and continuous distribution ( Figure 1A ). In the full cohort, caffeine levels correlated to metabolic changes in metabolism, including a negative association with branched chain amino acids (Valine, Isoleucine, Threonine) and the related carnitine-conjugated ketoacid (AC 5-OH), with positive association with the bacterial metabolites 3-sulfocatechol and Hippurate, kynurenine metabolites (8-methoxykynurenate), and lipid oxidation products (prostaglanding A2/B2 isomers, several acyl-carnitines (AC – Figure 1B ). Download figure Open in new tab Fig. 1 – Caffeine, donor biology and red blood cell (RBC) hemolysis in 13,091 REDS RBC Omics donors Caffeine distribution in 13,091 packed RBC units from donors enrolled in the REDS RBC Omics study ( A ). In B , metabolic correlates to caffeine levels in the same blood units. In C , caffeine levels were reproducible across multiple independent donations 6-12 months apart from 643 Index donors who were also enrolled in the Recalled phase of the study, owing to their extreme hemolytic propensity (95 th percentile). In D , stratification of REDS Index donors based on caffeine levels (top and bottom 1,000 donors by high or low caffeine levels) as a function of donor sex, age, body mass index (BMI), self-reported ethnicity and blood center in which the donors enrolled. In E , the volcano plot highlight the significant metabolic changes between caffeine low and high donors (n=1,000 per group – ATP is highlighted in the violin plot in the bottom right corner) in REDS Index donors at storage day 42. In the recalled donor cohort (n=643), caffeine levels did not change in packed RBCs when dosed at storage day 10, 23 and 42 (indent line plot). In F , blood units with high caffeine levels were associated with significantly higher storage ad osmotic hemolysis (** p<0.01; **** p<0.0001). For a subset of donors (extreme hemolyzers) enrolled both in the Index and Recalled phase of the REDS RBC Omics study (n=643), caffeine levels in packed RBCs were reproducible across multiple donations 6–12 months apart ( Figure 1C ). Stratification of donors based on high vs low caffeine levels (n=1,000 per group) revealed interesting demographic trends, with higher levels dosed in older individuals, females under 51, and donors with lower BMI, especially donors of Caucasian descent enrolled by the American Red Cross ( Figure 1D ). Similar trends were observed for the primary caffeine metabolite, paraxanthine (80% of caffeine is metabolized to paraxanthine), which was higher in older donors or younger females of Caucasian descent, showing a strong positive correlation (0.7 Spearman) and stable ratios across donor ages and sex ( Supplementary Figure 1A-C ). However, paraxanthine to caffeine ratios, a proxy for caffeine metabolism, were tendentially higher in younger donors, especially females of African American, Asian and Hispanic descent ( Supplementary Figure 1A-D ). Metabolite quantitative trait loci (mQTL) analysis identified a region on chromosome 19 coding for cytochrome p450 2A6 (CYP2A6) associated with paraxanthine and paraxanthine/caffeine levels in the REDS Index cohort ( Supplementary Figure 2A-B ). Top SNPs included rs7251570, rs76112798 and rs11667314, which were most prevalent in donors of Asian or European descent and significantly influenced paraxanthine metabolism ( Supplementary Figure 2C-D ).). Similar findings were recapitulated for the second most abundant caffeine metabolite (12% of total), theobromine, also mapping on the CYP2A6 region ( Supplementary Figure 3A-B ). Caffeine levels in REDS donors or coffee consumption in volunteers is associated with altered metabolism and increased hemolysis Comparative metabolomics between donors with the highest and lowest RBC caffeine levels identified significant alterations in central metabolism, including reduced glycolytic intermediates, increased purine catabolites, and changes in amino acid and redox pathways ( Figure 1.E ). To isolate the impact of caffeine on the circulating metabolome, we performed metabolomics analyses on plasma and RBCs from eight volunteers after consuming a cup of coffee (chemex or espresso – Supplementary Figure 4 ). Results showed transient elevation of circulating caffeine within 45 min, and rapid metabolism by 300 min. Elevation in sulfocatechol and other bacterial metabolites was observed in plasma and RBCs, consistent with caffeine’s impact on the gut microbiome. Caffeine consumption was associated with a drop in RBC 2,3-BPG and an increase in markers of membrane lipid oxidation and remodeling, including free fatty acids, (hydroxy)acyl-carnitines and lipid hydroperoxides (20:4 HPETE – Supplementary Figure 4 ). Elevation of plasma 2,3-BPG over time suggested release from RBCs, possibly via minor hemolysis. In the Recalled cohort (n=643), for which samples were collected at storage days 10, 23, and 42, caffeine levels remained stable ( Figure 1E -inset ), indicating that associated metabolic changes were not due to caffeine degradation. High caffeine levels correlated with significantly lower ATP and pentose phosphate pathway intermediates, suggesting impaired energy/redox metabolism and increased hemolytic propensity. Indeed, higher spontaneous and osmotic hemolysis at day 42 were observed in the high caffeine group (p<0.01 and p<0.0001, respectively – Figure 1F ). RBCs with high caffeine show redox imbalance, poor metabolic fitness, and impaired post-transfusion efficacy Metabolomics, proteomics, and lipidomics analyses of RBCs from Recalled donors (n=100) revealed subtle but consistent separation by caffeine levels in 3D uMAP projections ( Figure 2A–B ). High caffeine levels tracked with increased donor age and were enriched in women under 51 ( Figure 2C ). Longitudinal metabolomics revealed sustained increases in hypoxanthine and kynurenine ( Figure 2D–E ), markers of oxidative stress previously associated with poor PTR. Higher osmotic fragility was confirmed ( Figure 2F ). Hive plot correlation analyses revealed altered metabolic networks in high caffeine units, with weaker associations between ATP or 2,3-BPG and hemolysis ( Figure 2G ), metabolites previously linked to better PTR and oxygen off-loading ( 8 , 27 – 30 ). Download figure Open in new tab Fig. 2 – Higher caffeine levels are associated with elevated osmotic fragility of 643 recalled donors packed RBC and lower hemoglobin increments in transfusion recipients The 3D uMAP in A shows substantial overlap between packed RBCs with high or low caffeine (n=100 per group – distributions in B ) as a function of storage duration (z axis). Caffeine was higher in donors older than 30, while higher caffeine levels were observed in female donors below the age of 51 compared to males ( C ). In D-E , blood units with elevated caffeine show significantly higher levels of metabolic markers of poor post-transfusion recovery, hypoxanthine (p<0.05 at storage day 42) and kynurenine (p<0.01 through the whole storage period). Recalled donor blood units with high caffeine showed significantly higher levels of osmotic hemolysis ( F ). In G , hive plot summarizes changes in correlation networks of low and high caffeine blood units to hemolysis parameters (spontaneous storage hemolysis, or after osmotic or oxidant stress), and to functional metabolic parameters, including adenosine triphosphate (ATP) and 2,3-bisphosphoglycerate (2,3-BPG). Upomn interrogation of the REDS vein-to-vein database ( H ), transfusion of packed RBCs with high caffeine (or from donors repeatedly identified as heavy caffeine consumers – highest quartile) results in lower hemoglobin increments after transfusion, especially when the donors are also carriers of one or two alternative alleles for the single nucleotide polymorphism (SNP) for the adenosine receptor ADORA2b AX-13163261 ( I ). Similarly, transfusion of units from donors carrying 1 or 2 alleles for the ADORA2b rs758857 SNP was associated with higher delta-bilirubin increments, independently of caffeine levels ( J ). In vitro incubation of recombinantly expressed canonical or African G6PD variant (V68M; N126D) with 50 µM caffeine inhibited canonical but not deficient G6PD activity ( K ). In L , heat map of the top 50 significant differences between high and low caffeine level blood units in REDS RBC omics Index donors identifies donor age, markers of osmotic fragility (kynurenine), markers of lipid peroxidation (hydroxyacyl-carnitines and prostaglandin F2α) and alternative alleles for ADORA2b SNP rs758857 as the most significant differences between the two groups (high and low caffeine – n=1,000). Transfusion data from the REDS vein-to-vein database ( Figure 2H ) showed significantly lower hemoglobin increments in recipients of high-caffeine RBC units, particularly among donors carrying the ADORA2B SNP AX-13163261 ( Figure 2I ). The ADORA2B SNP rs758857 was also associated with higher increases in bilirubin post-transfusion, a hemolysis marker, independent of donor age or pre-transfusion bilirubin levels (n=1,598 - Figure 2J , Supplementary Figure 5 ). In vitro, 50 µM caffeine inhibited recombinant G6PD activity but did not further decrease the activity of the African variant (V68M; N126D), suggesting partial inhibition may explain metabolic defects ( Figure 2K ). Integrative omics and genomic analyses highlighted differences in membrane stability and ADORA2B variant prevalence among the top 50 discriminants between high and low caffeine donors ( Figure 2L ). ADORA2b genetic variants are prevalent and associate with increased RBC hemolysis We analyzed 879,000 SNPs in the REDS cohort focusing on ADORA2B. Several variants were common, including AX-13163261 (46.9% heterozygous, 19.9% homozygous – Figure 3A–B ). Variants rs758857 and AX-13163261 were strongly associated with osmotic (p<e-50) and oxidative hemolysis (p<e-9 - Figure 3C ), though not storage hemolysis. These alleles were enriched in older donors (rs72821748), those with higher BMI (rs758857), and African Americans ( Figure 3D–F , Supplementary Figure 6 ). Metabolomics linked these SNPs to altered redox metabolism and elevated caffeine levels ( Figure 3G–I ). Download figure Open in new tab Fig. 3 – Prevalence of ADORA2b SNPs in REDS Index donors and association with hemolytic propensity A list of the ADORA2b SNPs assayed in the 13,091 REDS Index donors is summarized in A , and the prevalence of heterozygosity or homozygosity for the alternative alleles is show in B . In C , SNP association to osmotic (blue), oxidative (red) or storage hemolysis (green). In D-F , stratification of the ADORA2b SNPs most strongly associated with hemolysis (especially osmotic and oxidative) by donor demographics (sex and ethnicity; age and demographics are shown in Supplementary Figure 1 ). In G-I , volcano plots of Spearman correlation of allele copies to metabolites and hemolysis parameters in the same SNPs. ADORA2b knockout impairs glycolysis and redox homeostasis in stored murine RBCs To model mechanisms, we stored RBCs from WT and ADORA2B KO mice for 0, 7, or 12 days (equivalent to human 0, 21, 42 days – Figure 4A-B ). Untargeted metabolomics showed separation by genotype and storage duration ( Figure 4C ). Combined with tracing data from 1,2,3- 13 C 3 -glucose experiments, our results confirmed impaired glycolytic flux in KO RBCs ( Figure 4D ), with reductions in intermediates of glycolysis, PPP, and glutathione metabolism ( Figure 4E , Figure 5 ). KO RBCs had lower adenylate pools (calculated as ATP+0.5 ADP/(ATP+ADP+AMP)) and altered GSH/GSSG ratios, consistent with oxidative stress seen in high-caffeine RBCs. Download figure Open in new tab Fig. 4 – ADORA2b regulates glycolytic fluxes in stored RBCs A literature-based summary model of the role of ADORA2b in regulating RBC metabolic fluxes through glycolysis based on Liu et al. Circulation 2016 ( A ). In B , packed RBCs from WT C57BL6/J mice (n=3) and ADORA2b KO mice (n=3) were stored for 0, 7, 12 days (equivalent to human RBC storage for 42 days) under refrigerated conditions in standard storage solutions, prior to metabolomics analysis. In C , principal component analysis of metabolomics data from this study shows a significant effect of storage duration and genotype. In D , the experiment was repeated by supplementing CPDA1 with 5 mM 1,2,3- 13 C 3 -glucose, which affords the direct measurements of metabolic fluxes through glycolysis by measuring the 13 C 3 -lactate isotopologue. Results indicate a significant decrease in 13 C 3 -lactate in ADORA2b KO mice. In E , heat map of the top 50 metabolites by time series two-way ANOVA (by genotype and storage) in WT vs ADORA2b KO mice at storage day 0, 7 and 12. Download figure Open in new tab Fig. 5 – Overview of glycolysis, the pentose phosphate pathway, glutathione and purine metabolism in stored WT and ADORA2b KO mice Red lines: ADORA2b; Blue lines: WT; n=3 per group, median + ranges (solid line and faded band) are shown for each metabolite. Abbreviations: GLUC: glucose; HexP: hexose phosphate (isomers); FBP: fructose bisphosphate; G3P: glyceraldehyde 3-phosphate; PGLY: phosphoglycerate (isomers); PEP: phosphoenol-pyruvate; PYR: pyruvate; LAC: lactate; PGLUC: 6-phosphogluconage; PentP: pentose phosphate (isomers); GSH and GSSG: reduced and oxidized glutathione; AXP: adenylate pool (ATP + 0.5 ADP / (ATP+ADP+AMP). Loss of ADORA2b alters redox-sensitive proteins and proteostasis in stored RBCs We profiled the RBC proteome of WT and KO mice across storage (days 0, 7, 12 – Figure 6A ). PCA and LDA confirmed genotype-dependent differences ( Figure 6B–C ). Time-series ANOVA identified changes in redox regulators (Psmd5, Ube2l3), glycolysis enzymes (Pkm), cytoskeletal proteins (Tubb4a, Dmtn, Flna), and vesiculation markers (Cltc – Figure 6D ). Redox proteomics revealed increasing oxidation over time in KO RBCs ( Figure 6E ). Pathway analysis implicated dysregulated proteostasis and antioxidant defenses ( Figure 6F ). Download figure Open in new tab Fig. 6 – Proteomics of stored RBCs from WT and ADORA2b knockout In A , overview of the experimental design. Principal component analysis ( B ) and linear discriminant analysis ( C ) of proteomics results based on genotype and storage duration (0, 7 or 12 days). In D , heat map of the top 50 significant proteins (official gene symbols) by two-way time series ANOVA as a function of ADORA2b KO and storage duration. In E , top 25 sulfenic, sulfonic or cysteine to dehydroalanine redox modifications between WT and ADORA2b KO RBCs during storage. In F , pathway analysis based on proteomics results. ADORA2b deletion impairs PTR in stored RBCs, an effect exacerbated by caffeine PTR studies in GFP+ recipient mice transfused with WT or KO RBCs stored 7 or 12 days showed reduced recovery in KO RBCs ( Figure 7A–B ). PTR positively correlated with ATP and glycolytic intermediates ( Figure 7C–D ). Storage with 100 µM caffeine further reduced PTR in KO but not WT RBCs ( Figure 7F ), demonstrating a gene–environment interaction. Clustering of top caffeine- and storage-sensitive metabolites emphasized disruptions in redox and nucleotide metabolism in KO RBCs ( Figure 7E ). A mechanistic model is presented in Figure 7G . Download figure Open in new tab Fig. 7 – ADORA2b KO negatively impacts post-transfusion recovery (PTR) of stored RBCs and is aggravated by caffeine supplementation PTR studies were performed on stored (day 7 and 12) RBCs from WT and ADORA2b KO mice through transfusion into Ubi-GFP mice and co-transfusion with test mCherry fresh WT RBCs ( A ). Results indicate a significant drop in PTR in KO mice ( B ). In C , metabolic correlates to PTR. Supplementation of 100 µM caffeine to stored RBCs from WT and ADORA2b KO mice significantly impacted RBC metabolism (top 50 metabolites by two-way ANOVA are shown in the heat map in D ). Highlights include cytidine monophosphate (CMP) and ATP ( E ). Caffeine supplementation further decreased PTR in ADORA2b KO mice ( F ). A working model is proposed in G , based on experimental data and existing literature on the role of caffeine as an ADORA2b antagonist and G6PD inhibitor. Adenosine supplementation only partially restored metabolic defects in human stored RBCs Given the role of adenosine signaling, we tested adenosine supplementation (1, 5, 10 µM) in stored human RBCs ( Supplementary Figure 7 ). Adenosine increased intracellular levels early in storage but had modest metabolic impact. Lactate was reduced; ATP, ADP, and 2,3-BPG were higher in the first week. After three weeks, AMP, IMP, and inosine increased in supplemented units, consistent with adenosine catabolism and limited long-term efficacy ( Supplementary Figure 7D–E ). DISCUSSION In the present study we report that caffeine, a ubiquitous dietary component, is a previously underappreciated factor influencing the quality of stored RBCs. By leveraging a large-scale cohort of more than 13,000 blood donors from the REDS RBC-Omics study, we demonstrate that caffeine levels – detected in blood products at biologically relevant thresholds in the micromolar to hundreds of micromolar range ( 18 ) - correlate strongly with impaired RBC metabolic integrity and increased storage hemolysis, translating into decreased transfusion efficacy. Stratification of donors by circulating caffeine levels revealed demographic patterns consistent with established population-level consumption trends. Epidemiological surveys in the United States and Europe report higher average caffeine intake among older adults, largely attributable to habitual coffee consumption, which increases with age and plateaus in the 60s ( 31 ). Although men typically consume slightly more caffeine overall, studies suggest that women—particularly those under 51—may exhibit higher circulating caffeine concentrations due to slower metabolism influenced by hormonal factors, including estrogen and oral contraceptive use ( 31 ). Our measurements of paraxanthine/caffeine ratios are consistent with this model. Caffeine levels also tend to be higher in individuals with lower BMI, likely reflecting a smaller volume of distribution and possibly differences in metabolic clearance. Ethnic variation in both consumption habits and metabolism contributes further: non-Hispanic White individuals report the highest coffee intake in national surveys, a pattern reflected in our cohort, while inter-individual variation in CYP1A2 genotype and expression may additionally shape plasma caffeine levels ( 32 ). Collectively, these findings support the interpretation that lifestyle, demographic, and pharmacogenetic factors converge to influence measurable caffeine exposure in blood donors. While caffeine effects on the central nervous system, cardiovascular disease and cancer ( 33 ) are well documented, peripheral actions—especially on mature erythrocytes— have remained underexplored. Here, we show that elevated caffeine in RBC units is consistently associated with a metabolic phenotype indicative of compromised glycolytic activity, reduced ATP, total adenylate pools and 2,3-BPG, as well as with heightened oxidative stress, ferroptosis-like processes ( 34 ) and osmotic fragility markers, including oxidized glutathione ( 35 ), lipid peroxidation products ( 34 ) and hydroxyacyl-carnitines ( 36 ), hypoxanthine ( 8 ) and kynurenine ( 28 ). These findings were validated in volunteers within 45 min to 5h after consuming coffee, independently from brewing methods (espresso vs chemex). These biochemical alterations culminate in increased spontaneous and stress-induced hemolysis during storage, consistent with the increased susceptibility to extravascular hemolysis via splenic sequestration and erythrophagocytosis ( 37 ) of storage-damaged RBCs that become energetically effete ( 38 ) and accumulate oxidant damage, leading to elevated proteasomal degradation ( 39 , 40 ). Clinically relevant, recipients of RBC units from donors with high caffeine consumption experienced significantly lower post-transfusion hemoglobin increments. Importantly, these effects were more pronounced in donors harboring specific polymorphisms in the ADORA2b gene, a receptor expressed on RBC membranes for which caffeine is a competitive antagonist, illustrating a critical gene-exposome interaction ( 17 ). Observations in human RBCs are mechanistically supported by our parallel studies in a murine model deficient in ADORA2b ( 41 ). While adenosine signaling via ADORA2b signaling has been previously linked to acclimatization to high altitude hypoxia by favoring energy metabolism to boost oxygen off-loading ( 4 ), our findings here underscore the critical role of purinergic signaling in maintaining RBC metabolism during refrigerated storage. Inhibition or genetic deletion of ADORA2b disrupts this metabolic adaptation, mirroring the metabolic dysregulation observed with caffeine exposure. In line with this, ADORA2b knockout mice exhibited markedly diminished glycolytic flux, reduced ATP levels, and impaired glutathione recycling during RBC storage, phenocopying the human high-caffeine RBC phenotype. The subsequent decline in RBC viability post-transfusion in knockout mice was further exacerbated by caffeine supplementation during storage, confirming the synergistic detriment of ADORA2b inhibition and caffeine exposure. Notably, our results confirms and expands upon previous hypotheses implicating caffeine as a direct inhibitor of G6PD ( 15 ). Our direct enzymatic assays reveal inhibitory effects of caffeine on canonical G6PD activity at physiologically relevant concentrations. However, the effect did not further compound with G6PD deficiency in the African variant. Therefore, the adverse effects of caffeine observed in stored RBC units are potentially also direct, mediated both through disrupted purinergic signaling (ADORA2b) and direct enzymatic inhibition (G6PD), in part mimicking the increased susceptibility to hemolysis and impaired post-transfusion survival of RBCs from G6PD deficient donors( 24 , 26 , 35 , 42 ). The translational implications of our findings are significant. First, donor caffeine consumption—a common dietary exposure for up to 75% of Americans—emerges as a modifiable behavioral factor potentially influencing RBC storage quality and transfusion outcomes. Given caffeine’s short biological half-life, transient dietary modifications around the time of blood donation might mitigate its negative impact, aligning with blood donation guidelines in several European countries where donors are advised to limit caffeine intake prior to donation. Conversely, in other regions, such as the United States, caffeine consumption before blood donation is not actively discouraged and may even be implicitly encouraged due to its known beneficial acute effects on blood pressure, potentially expediting the donation process and reducing vasovagal reactions. Indeed, moderate caffeine intake can transiently increase donor blood pressure and vascular tone, facilitating venous access and blood withdrawal efficiency. However, this advantage must be balanced against caffeine’s mild diuretic properties, which may predispose donors to dehydration—an established risk-factor for adverse donation-related events and poorer blood flow during collection. Beyond caffeine, our findings advocate for incorporating donor exposome data—dietary habits, medication usage—along with genetic markers into precision transfusion medicine strategies aimed at informing blood inventory management. For example, RBC units from high-caffeine-consuming donors or individuals with high-risk ADORA2b polymorphisms might be preferentially allocated to scenarios where slight variations in RBC quality are clinically less impactful. Conversely, units from donors with low caffeine exposure and favorable genetic profiles could be prioritized for high-risk transfusion recipients, such as neonates or patients with severe anemia. Moreover, our mechanistic insights point to possible pharmacological interventions aimed at bolstering ADORA2b signaling or downstream metabolic pathways during RBC storage. Adenosine supplementation to standard storage additives transiently boosted energy metabolism, leading to early increases in intracellular adenosine and total adenylate pools, followed by elevation of purine deamination products inosine and IMP after storage week 3. Altogether, these results are consistent with adenosine uptake via ENT1, rather than activation of ADORA2b, followed by deamination of adenosine itself or its metabolite AMP into inosine and IMP by adenosine deaminase ( 27 ) or RBC-specific AMP deaminase 3 ( 43 ) Degradation of adenosine supplements in storage mediate would thus only favor transient stimulatory effects of ADORA2b, while the absence in stored RBC units of hepatic CYP2As that can degrade caffeine would not similarly constraint the antagonistic effects on the same receptors. In light of these results, alternative strategies to stimulate ADORA2b might include non-degradable adenosine analogs or allosteric modulators specifically enhancing glycolysis and redox balance, thereby improving stored RBC resilience and transfusion efficacy. Preliminary data from models utilizing AMPK activators ( 13 ) or hypoxic storage ( 25 , 44 ) conditions provide proof-of-concept for these approaches, highlighting the feasibility of metabolic modulation as a storage enhancement strategy. Corollary to these considerations, cytochrome p450 reductases are NADPH dependent enzymes.( 45 ) As such, direct G6PD inhibition by caffeine may constraint the pools of the very NADPH necessary for its direct catabolism or that of its metabolites like paraxanthine. Indeed, our mQTL analysis here identified an association between donor genetics (CYP2A6 SNPs, especially in donors of Asian and European descent) and paraxanthine metabolism, consistent with the literature ( 46 , 47 ). While our study benefits from robust epidemiological evidence combined with detailed mechanistic validation, we acknowledge certain limitations. The modest effect sizes observed in clinical transfusion outcomes raise questions about the absolute clinical relevance. Nevertheless, given the substantial global volume of transfusions, even small improvements in RBC quality can translate into meaningful clinical benefits at the population level. Additionally, our animal model, though highly informative, utilized defined genetic modifications; thus, translating these findings to routine clinical settings demands further validation. Collectively, our study positions caffeine consumptionas a significant, modifiable factor influencing RBC metabolic responses to oxidant stress, here in the form of a common iatrogenic intervention such as storage under blood bank conditions. Our data thus suggests that caffeine might sensitize RBCs to hemolysis by osmotic or oxidant insults, common stressors for example in response to moderate ( 48 ) or strenuous exercise ( 49 ). It also underscores the importance of the gene-exposome interface in determining transfusion outcomes, advocating for precision approaches in transfusion medicine. Future investigations should further explore dietary guidance for donors, pharmacological enhancements of RBC metabolic pathways, and more comprehensive integration of donor metadata into clinical transfusion practices. Funding AD and JCZ were supported by funds by the National Heart, Lung, and Blood Institute (NHLBI) (R01HL146442, R01HL149714). The REDS RBC Omics and REDS-IV-P CTLS programs are sponsored by the NHLBI contract 75N2019D00033, and from the NHLBI Recipient Epidemiology and Donor Evaluation Study-III (REDS-III) RBC Omics project, which was supported by NHLBI contracts HHSN2682011-00001I, −00002I, −00003I, −00004I, −00005I, −00006I, −00007I, −00008I, and −00009I. G.R.K was supported by grants from the National Institute of General Medical Sciences (NIGMS), F32GM124599. NR received funding from NHLBI (R01HL126130). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. The authors would like to thank all the donor volunteers who participated in this study and all the global blood donor communities for their life-saving altruistic gifts. Author Contribution Animal studies: AH, JCZ. Proteomics: MD, KCH. Metabolomics and lipidomics analyses: DS, TN, AD. Biostatistics and Bioinformatics: GRK, XD, AD. REDS RBC Omics: MS, SK, SLS, PJN, MPB. Vein-to-vein database: NR; Human genetics: GRK, ALM, GPP (human). Figure preparation: AD. Conceptualization: YX, JCZ, AD. Writing and finalization: first draft by AD and all co-authors reviewed and approved the final version. Competing Interest The authors declare that AD, KCH, TN are founders of Omix Technologies Inc. AD and TN are Scientific Advisory Board (SAB) members for Hemanext Inc. AD is SAB member for Macopharma Inc and SynthMed Bio. All the other authors have no conflicts to disclose in relation to this study. Data and Materials availability All raw data and elaborations are included in Supplementary Table 1.xlsx. ADORA2b KO mice are available upon reasonable request, finalization of material transfer agreement and after institutional ACUC approval through Dr James C Zimring Lab at the University of Virginia ( jcz2k{at}virginia.edu ). Further information and requests for resources and reagents should be directed to and will be fulfilled by the Lead Contact, Angelo D’Alessandro ( angelo.dalessandro{at}cuanschutz.edu ). Data analysis and Statistical analyses – including hierarchical clustering analysis (HCA), linear discriminant analysis (LDA), uniform Manifold Approximation and Projection (uMAP), correlation analyses and Lasso regression were performed using both MetaboAnalyst 5.0 and in-house developed code in RStudio (2024.12.1 Build 563). LIST of SUPPLEMENTARY MATERIALS Supplementary Materials.pdf includes: SUPPLEMENTARY MATERIALS AND METHODS EXTENDED SUPPLEMENTARY FIGURES SUPPLEMENTARY FIGURE 1 SUPPLEMENTARY FIGURE 2 SUPPLEMENTARY FIGURE 3 SUPPLEMENTARY FIGURE 4 SUPPLEMENTARY FIGURE 5 SUPPLEMENTARY FIGURE 6 Supplementary table 1.xlsx – including all raw data and elaborations. Acknowledgements Funder Information Declared National Heart Lung and Blood Institute, https://ror.org/012pb6c26 , R01HL146442 , R01HL149714 , 75N2019D00033 , HHSN2682011 , R01HL126130 National Institute of General Medical Sciences, https://ror.org/04q48ey07 , F32GM124599 REFERENCES 1. ↵ Temple JL , Bernard C , Lipshultz SE , Czachor JD , Westphal JA , Mestre MA . The Safety of Ingested Caffeine: A Comprehensive Review . Front Psychiatry . 2017 ; 8 : 80 . eng. Epub 20170526. doi: 10.3389/fpsyt.2017.00080 . Cited in: Pubmed; PMID 28603504 . OpenUrl CrossRef PubMed 2. ↵ Cappelletti S , Piacentino D , Sani G , Aromatario M . Caffeine: cognitive and physical performance enhancer or psychoactive drug? Curr Neuropharmacol . 2015 Jan ; 13 ( 1 ): 71 – 88 . eng. doi: 10.2174/1570159×13666141210215655 . Cited in: Pubmed; PMID 26074744 . OpenUrl CrossRef PubMed 3. ↵ Ribeiro JA , Sebastião AM. Caffeine and adenosine . J Alzheimers Dis . 2010 ; 20 Suppl 1 : S3 - 15 . eng. doi: 10.3233/jad-2010-1379 . Cited in: Pubmed; PMID 20164566 . OpenUrl CrossRef PubMed 4. ↵ Liu H , Zhang Y , Wu H , D’Alessandro A , Yegutkin GG , Song A , Sun K , Li J , Cheng NY , Huang A , Edward Wen Y , Weng TT , Luo F , Nemkov T , Sun H , Kellems RE , Karmouty-Quintana H , Hansen KC , Zhao B , Subudhi AW , Jameson-Van Houten S , Julian CG , Lovering AT , Eltzschig HK , Blackburn MR , Roach RC , Xia Y . Beneficial Role of Erythrocyte Adenosine A2B Receptor-Mediated AMP-Activated Protein Kinase Activation in High-Altitude Hypoxia . Circulation . 2016 Aug 2 ; 134 ( 5 ): 405 – 21 . doi: 10.1161/CIRCULATIONAHA.116.021311 . Cited in: Pubmed; PMID 27482003 . OpenUrl Abstract / FREE Full Text 5. ↵ Sun K , D’Alessandro A , Xia Y . Purinergic control of red blood cell metabolism: novel strategies to improve red cell storage quality . Blood Transfus . 2017 Oct ; 15 ( 6 ): 535 – 542 . Epub 20170412. doi: 10.2450/2017.0366-16 . Cited in: Pubmed; PMID 28488967 . OpenUrl CrossRef PubMed 6. ↵ Donovan K , Meli A , Cendali F , Park KC , Cardigan R , Stanworth S , McKechnie S , D’Alessandro A , Smethurst PA , Swietach P . Stored blood has compromised oxygen unloading kinetics that can be normalized with rejuvenation and predicted from corpuscular side-scatter . Haematologica . 2022 Jan 1 ; 107 ( 1 ): 298 – 302 . Epub 2021/09/10. doi: 10.3324/haematol.2021.279296 . Cited in: Pubmed; PMID 34498445 . OpenUrl CrossRef PubMed 7. ↵ Yoshida T , Prudent M , D’Alessandro A . Red blood cell storage lesion: causes and potential clinical consequences . Blood Transfus . 2019 Jan ; 17 ( 1 ): 27 – 52 . Epub 2019/01/18. doi: 10.2450/2019.0217-18 . Cited in: Pubmed; PMID 30653459 . OpenUrl CrossRef PubMed 8. ↵ Nemkov T , Stephenson D , Earley EJ , Keele GR , Hay A , Key A , Haiman ZB , Erickson C , Dzieciatkowska M , Reisz JA , Moore A , Stone M , Deng X , Kleinman S , Spitalnik SL , Hod EA , Hudson KE , Hansen KC , Palsson BO , Churchill GA , Roubinian N , Norris PJ , Busch MP , Zimring JC , Page GP , D’Alessandro A . Biological and genetic determinants of glycolysis: Phosphofructokinase isoforms boost energy status of stored red blood cells and transfusion outcomes . Cell Metabolism . 2024 . doi: 10.1016/j.cmet.2024.06.007 . OpenUrl CrossRef 9. ↵ Rogers SC , Ge X , Brummet M , Lin X , Timm DD , d’Avignon A , Garbow JR , Kao J , Prakash J , Issaian A , Eisenmesser EZ , Reisz JA , D’Alessandro A , Doctor A . Quantifying dynamic range in red blood cell energetics: Evidence of progressive energy failure during storage . Transfusion . 2021 ; 61 ( 5 ): 1586 – 1599 . doi: 10.1111/trf.16395 . OpenUrl CrossRef 10. ↵ Peltier S , Marin M , Dzieciatkowska M , Dussiot M , Roy MK , Bruce J , Leblanc L , Hadjou Y , Georgeault S , Fricot A , Roussel C , Stephenson D , Casimir M , Sissoko A , Paye F , Dokmak S , Ndour PA , Roingeard P , Gautier EF , Spitalnik SL , Hermine O , Buffet PA , D’Alessandro A , Amireault P. Proteostasis and metabolic dysfunction in a distinct subset of storage-induced senescent erythrocytes targeted for clearance . bioRxiv . 2024 Sep 12 . eng. Epub 20240912. doi: 10.1101/2024.09.11.612195 . Cited in: Pubmed; PMID 39314353 . OpenUrl Abstract / FREE Full Text 11. ↵ D’Alessandro A , Nemkov T , Sun K , Liu H , Song A , Monte AA , Subudhi AW , Lovering AT , Dvorkin D , Julian CG , Kevil CG , Kolluru GK , Shiva S , Gladwin MT , Xia Y , Hansen KC , Roach RC . AltitudeOmics: Red Blood Cell Metabolic Adaptation to High Altitude Hypoxia . J Proteome Res . 2016 Oct 7 ; 15 ( 10 ): 3883 – 3895 . doi: 10.1021/acs.jproteome.6b00733 . Cited in: Pubmed; PMID 27646145 . OpenUrl CrossRef PubMed 12. ↵ Zhang Y , Dai Y , Wen J , Zhang W , Grenz A , Sun H , Tao L , Lu G , Alexander DC , Milburn MV , Carter-Dawson L , Lewis DE , Zhang W , Eltzschig HK , Kellems RE , Blackburn MR , Juneja HS , Xia Y . Detrimental effects of adenosine signaling in sickle cell disease . Nat Med . 2011 Jan ; 17 ( 1 ): 79 – 86 . eng. Epub 20101219. doi: 10.1038/nm.2280 . Cited in: Pubmed; PMID 21170046 . OpenUrl CrossRef PubMed 13. ↵ Foretz M , Guihard S , Leclerc J , Fauveau V , Couty J-P , Andris F , Gaudry M , Andreelli F , Vaulont S , Viollet B . Maintenance of red blood cell integrity by AMP-activated protein kinase α1 catalytic subunit . FEBS Letters . 2010 2010/08/20/; 584 ( 16 ): 3667 - 3671 . doi: 10.1016/j.febslet.2010.07.041 . OpenUrl CrossRef PubMed 14. ↵ Song A , Zhang Y , Han L , Yegutkin GG , Liu H , Sun K , D’Alessandro A , Li J , Karmouty-Quintana H , Iriyama T , Weng T , Zhao S , Wang W , Wu H , Nemkov T , Subudhi AW , Jameson-Van Houten S , Julian CG , Lovering AT , Hansen KC , Zhang H , Bogdanov M , Dowhan W , Jin J , Kellems RE , Eltzschig HK , Blackburn M , Roach RC , Xia Y . Erythrocytes retain hypoxic adenosine response for faster acclimatization upon re-ascent . Nat Commun . 2017 Feb 7 ; 8 : 14108 . eng. Epub 2017/02/09. doi: 10.1038/ncomms14108 . Cited in: Pubmed; PMID 28169986 . OpenUrl CrossRef PubMed 15. ↵ Xu H , Hu L , Liu T , Chen F , Li J , Xu J , Jiang L , Xiang Z , Wang X , Sheng J . Caffeine Targets G6PDH to Disrupt Redox Homeostasis and Inhibit Renal Cell Carcinoma Proliferation . Front Cell Dev Biol . 2020 ; 8 : 556162 . eng. Epub 20201006. doi: 10.3389/fcell.2020.556162 . Cited in: Pubmed; PMID 33123534 . OpenUrl CrossRef PubMed 16. ↵ D’Alessandro A . Red blood cell metabolism: a window on systems health towards clinical metabolomics . Curr Opin Hematol . 2025 May 1 ; 32 ( 3 ): 111 – 119 . eng. Epub 20250313. doi: 10.1097/moh0000000000000863 . Cited in: Pubmed; PMID 40085132 . OpenUrl CrossRef PubMed 17. ↵ Nemkov T , Stefanoni D , Bordbar A , Issaian A , Palsson BO , Dumont LJ , Hay A , Song A , Xia Y , Redzic JS , Eisenmesser EZ , Zimring JC , Kleinman S , Hansen KC , Busch MP , D’Alessandro A . Blood donor exposome and impact of common drugs on red blood cell metabolism . JCI Insight . 2021 Feb 8 ; 6 ( 3 ). Epub 20210208. doi: 10.1172/jci.insight146175 . Cited in: Pubmed; PMID 33351786 . OpenUrl CrossRef PubMed 18. ↵ D’Alessandro A , Fu X , Reisz JA , Kanias T , Page GP , Stone M , Kleinman S , Zimring JC , Busch M , Recipient E , Donor Evaluation S , III. Stored RBC metabolism as a function of caffeine levels . Transfusion . 2020 Jun ; 60 ( 6 ): 1197 – 1211 . Epub 2020/05/13. doi: 10.1111/trf15813 . Cited in: Pubmed; PMID 32394461 . OpenUrl CrossRef PubMed 19. ↵ Nemkov T , Yoshida T , Nikulina M , D’Alessandro A . High-Throughput Metabolomics Platform for the Rapid Data-Driven Development of Novel Additive Solutions for Blood Storage [Original Research] . Frontiers in Physiology . 2022 2022- March - 14 ; 13 . English. doi: 10.3389/fphys.2022.833242 . OpenUrl CrossRef 20. Nemkov T , Yoshida T , Nikulina M , D’Alessandro A . High-Throughput Metabolomics Platform for the Rapid Data-Driven Development of Novel Additive Solutions for Blood Storage . Front Physiol . 2022 ; 13 : 833242 . eng. Epub 2022/04/02. doi: 10.3389/fphys.2022833242 . Cited in: Pubmed; PMID 35360223 . OpenUrl CrossRef PubMed 21. ↵ Nemkov T , Reisz JA , Gehrke S , Hansen KC , D’Alessandro A . High-Throughput Metabolomics: Isocratic and Gradient Mass Spectrometry-Based Methods . Methods Mol Biol . 2019 ; 1978 : 13 – 26 . Epub 2019/05/24. doi: 10.1007/978-1-4939-9236-2_2 . Cited in: Pubmed; PMID 31119654 . OpenUrl CrossRef PubMed 22. ↵ Thomas T , Stefanoni D , Dzieciatkowska M , Issaian A , Nemkov T , Hill RC , Francis RO , Hudson KE , Buehler PW , Zimring JC , Hod EA , Hansen KC , Spitalnik SL , D’Alessandro A . Evidence for structural protein damage and membrane lipid remodeling in red blood cells from COVID-19 patients . medRxiv . 2020 Jun 30 . Epub 2020/07/09. doi: 10.1101/2020.06.2920142703 . Cited in: Pubmed; PMID 32637980 . OpenUrl Abstract / FREE Full Text 23. ↵ Reisz JA , Zheng C , D’Alessandro A , Nemkov T . Untargeted and Semi-targeted Lipid Analysis of Biological Samples Using Mass Spectrometry-Based Metabolomics . Methods Mol Biol . 2019 ; 1978 : 121 – 135 . Epub 2019/05/24. doi: 10.1007/978-1-4939-9236-2_8 . Cited in: Pubmed; PMID 31119660 . OpenUrl CrossRef PubMed 24. ↵ Page GP , Kanias T , Guo YJ , Lanteri MC , Zhang X , Mast AE , Cable RG , Spencer BR , Kiss JE , Fang F , Endres-Dighe SM , Brambilla D , Nouraie M , Gordeuk VR , Kleinman S , Busch MP , Gladwin MT . Multiple-ancestry genome-wide association study identifies 27 loci associated with measures of hemolysis following blood storage . J Clin Invest . 2021 Jul 1 ; 131 ( 13 ). eng. Epub 2021/05/21. doi: 10.1172/jci146077 . Cited in: Pubmed; PMID 34014839 . OpenUrl CrossRef PubMed 25. ↵ Hay A , Dziewulska K , Gamboni F , Nerguizian D , Dzieciatkowska M , Zimring JC , D’Alessandro A . Hypoxic storage of murine red blood cells improves energy metabolism and post-transfusion recoveries . Blood Transfus . 2022 Oct 21 . eng. Epub 2022/11/09. doi: 10.2450/2022.0172-22 . Cited in: Pubmed; PMID 36346885 . OpenUrl CrossRef PubMed 26. ↵ Roubinian NH , Reese SE , Qiao H , Plimier C , Fang F , Page GP , Cable RG , Custer B , Gladwin MT , Goel R , Harris B , Hendrickson JE , Kanias T , Kleinman S , Mast AE , Sloan SR , Spencer BR , Spitalnik SL , Busch MP , Hod EA . Donor genetic and nongenetic factors affecting red blood cell transfusion effectiveness . JCI Insight . 2022 Jan 11 ; 7 ( 1 ). eng. Epub 2021/11/19. doi: 10.1172/jci.insight.152598 . Cited in: Pubmed; PMID 34793330 . OpenUrl CrossRef PubMed 27. ↵ Nemkov T , Sun K , Reisz JA , Song A , Yoshida T , Dunham A , Wither MJ , Francis RO , Roach RC , Dzieciatkowska M , Rogers SC , Doctor A , Kriebardis A , Antonelou M , Papassideri I , Young CT , Thomas TA , Hansen KC , Spitalnik SL , Xia Y , Zimring JC , Hod EA , D’Alessandro A . Hypoxia modulates the purine salvage pathway and decreases red blood cell and supernatant levels of hypoxanthine during refrigerated storage . Haematologica . 2018 Feb ; 103 ( 2 ): 361 – 372 . eng. Epub 2017/10/29. doi: 10.3324/haematol.2017.178608 . Cited in: Pubmed; PMID 29079593 . OpenUrl Abstract / FREE Full Text 28. ↵ Nemkov T , Stephenson D , Erickson C , Dzieciatkowska M , Key A , Moore A , Earley EJ , Page GP , Lacroix IS , Stone M , Deng X , Raife T , Kleinman S , Zimring JC , Roubinian N , Hansen KC , Busch MP , Norris PJ , D’Alessandro A . Regulation of kynurenine metabolism by blood donor genetics and biology impacts red cell hemolysis in vitro and in vivo . Blood . 2024 Feb 1 ; 143 ( 5 ): 456 – 472 . doi: 10.1182/blood.2023022052 . Cited in: Pubmed; PMID 37976448 . OpenUrl CrossRef PubMed 29. Dumbill R , Rabcuka J , Fallon J , Knight S , Hunter J , Voyce D , Barrett J , Ellen M , Weissenbacher A , Kurniawan T , Blonski S , Korczyk PM , Ploeg R , Coussios C , Friend P , Swietach P . Impaired O2 unloading from stored blood results in diffusion-limited O2 release at tissues: evidence from human kidneys . Blood . 2024 Feb 22 ; 143 ( 8 ): 721 – 733 . doi: 10.1182/blood.2023022385 . Cited in: Pubmed; PMID 38048591 . OpenUrl CrossRef PubMed 30. ↵ D’Alessandro A , Earley EJ , Nemkov T , Stephenson D , Dzieciatkowska M , Hansen KC , Minetti G , Champigneulle B , Stauffer E , Pichon A , Furian M , Verges S , Kleinman S , Norris PJ , Busch MP , Page GP , Kaestner L . Genetic polymorphisms and expression of Rhesus blood group RHCE are associated with 2,3-bisphosphoglycerate in humans at high altitude . Proc Natl Acad Sci U S A . 2024 Jan 2 ; 121 ( 1 ): e2315930120 . eng. Epub 2023/12/26. doi: 10.1073/pnas.2315930120 . Cited in: Pubmed; PMID 38147558 . OpenUrl CrossRef PubMed 31. ↵ Mitchell DC , Knight CA , Hockenberry J , Teplansky R , Hartman TJ . Beverage caffeine intakes in the U.S . Food and Chemical Toxicology . 2014 2014/01/01/; 63 : 136 – 142 . doi: 10.1016/j.fct.2013.10.042 . OpenUrl CrossRef PubMed 32. ↵ Yang A , Palmer AA , de Wit H . Genetics of caffeine consumption and responses to caffeine . Psychopharmacology . 2010 2010/08/01; 211 ( 3 ): 245 – 257 . doi: 10.1007/s00213-010-1900-1 . OpenUrl CrossRef PubMed Web of Science 33. ↵ Dam RMv , Hu FB , Willett WC . Coffee, Caffeine, and Health . New England Journal of Medicine . 2020 ; 383 ( 4 ): 369 – 378 . doi : doi: 10.1056/NEJMra1816604 . OpenUrl CrossRef PubMed 34. ↵ D’Alessandro A , Keele GR , Hay A , Nemkov T , Earley EJ , Stephenson D , Vincent M , Deng X , Stone M , Dzieciatkowska M , Hansen KC , Kleinman S , Spitalnik SL , Roubinian N , Norris PJ , Busch MP , Page GP , Stockwell BR , Churchill GA , Zimring JC . Ferroptosis regulates hemolysis in stored murine and human red blood cells . Blood . 2025 Feb 13 ; 145 ( 7 ): 765 – 783 . eng. doi: 10.1182/blood.2024026109 . Cited in: Pubmed; PMID 39541586 . OpenUrl CrossRef PubMed 35. ↵ D’Alessandro A , Fu X , Kanias T , Reisz JA , Culp-Hill R , Guo Y , Gladwin MT , Page G , Kleinman S , Lanteri M , Stone M , Busch MP , Zimring JC , Recipient E , Donor Evaluation S , III. Donor sex, age and ethnicity impact stored red blood cell antioxidant metabolism through mechanisms in part explained by glucose 6-phosphate dehydrogenase levels and activity . Haematologica . 2021 May 1 ; 106 ( 5 ): 1290 – 1302 . Epub 2020/04/04. doi: 10.3324/haematol.2020.246603 . Cited in: Pubmed; PMID 32241843 . OpenUrl CrossRef PubMed 36. ↵ Nemkov T , Key A , Stephenson D , Earley EJ , Keele GR , Hay AM , Amireault P , Casimir M , Dussiot M , Dzieciatkowska M , Reisz JA , Deng X , Stone M , Kleinman SH , Spitalnik SL , Hansen KC , Norris PJ , Churchill GA , Busch MP , Roubinian NH , Page GP , Zimring JC , Arduini A , D’Alessandro A . Genetic regulation of carnitine metabolism controls lipid damage repair and aging RBC hemolysis in vivo and in vitro . Blood . 2024 Mar 21 . eng. Epub 20240321. doi: 10.1182/blood.2024023983 . Cited in: Pubmed; PMID 38513237 . OpenUrl CrossRef PubMed 37. ↵ Roussel C , Morel A , Dussiot M , Marin M , Colard M , Fricot-Monsinjon A , Martinez A , Chambrion C , Henry B , Casimir M , Volle G , Dépond M , Dokmak S , Paye F , Sauvanet A , Le Van Kim C , Colin Y , Georgeault S , Roingeard P , Spitalnik SL , Ndour PA , Hermine O , Hod EA , Buffet PA , Amireault P . Rapid clearance of storage-induced microerythrocytes alters transfusion recovery . Blood . 2021 Apr 29 ; 137 ( 17 ): 2285 – 2298 . eng. Epub 2021/03/04. doi: 10.1182/blood.2020008563 . Cited in: Pubmed; PMID 33657208 . OpenUrl CrossRef PubMed 38. ↵ van Wijk R , van Solinge WW . The energy-less red blood cell is lost: erythrocyte enzyme abnormalities of glycolysis . Blood . 2005 Dec 15 ; 106 ( 13 ): 4034 – 42 . eng. Epub 20050728. doi: 10.1182/blood-2005-04-1622 . Cited in: Pubmed; PMID 16051738 . OpenUrl Abstract / FREE Full Text 39. ↵ Tzounakas VL , Dzieciatkowska M , Anastasiadi AT , Karadimas DG , Vergaki A , Siourounis P , Stamoulis K , Papassideri IS , Kriebardis AG , D’Alessandro A , Antonelou MH . Red cell proteasome modulation by storage, redox metabolism and transfusion . Blood Transfus . 2022 Jan ; 20 ( 1 ): 27 – 39 . eng. Epub 2020/12/03. doi: 10.2450/2020.0179-20 . Cited in: Pubmed; PMID 33263521 . OpenUrl CrossRef PubMed 40. ↵ Peltier S , Marin M , Dzieciatkowska M , Dussiot M , Roy MK , Bruce J , Leblanc L , Hadjou Y , Georgeault S , Fricot A , Roussel C , Stephenson D , Casimir M , Sissoko A , Paye F , Dokmak S , Ndour PA , Roingeard P , Gautier EF , Spitalnik SL , Hermine O , Buffet PA , D’Alessandro A , Amireault P . Proteostasis and metabolic dysfunction characterize a subset of storage-induced senescent erythrocytes targeted for post-transfusion clearance . J Clin Invest . 2025 Mar 11 . eng. Epub 20250311. doi: 10.1172/jci183099 . Cited in: Pubmed; PMID 40067362 . OpenUrl CrossRef PubMed 41. ↵ Erblang M , Drogou C , Gomez-Merino D , Metlaine A , Boland A , Deleuze JF , Thomas C , Sauvet F , Chennaoui M . The Impact of Genetic Variations in ADORA2A in the Association between Caffeine Consumption and Sleep . Genes (Basel) . 2019 Dec 6 ; 10 ( 12 ). eng. The authors declare no conflict of interest. Epub 20191206. doi: 10.3390/genes10121021 . Cited in: Pubmed; PMID 31817803 . OpenUrl CrossRef PubMed 42. ↵ Francis RO , D’Alessandro A , Eisenberger A , Soffing M , Yeh R , Coronel E , Sheikh A , Rapido F , La Carpia F , Reisz JA , Gehrke S , Nemkov T , Thomas T , Schwartz J , Divgi C , Kessler D , Shaz BH , Ginzburg Y , Zimring JC , Spitalnik SL , Hod EA . Donor glucose-6-phosphate dehydrogenase deficiency decreases blood quality for transfusion . J Clin Invest . 2020 May 1 ; 130 ( 5 ): 2270 – 2285 . eng. Epub 2020/01/22. doi: 10.1172/jci133530 . Cited in: Pubmed; PMID 31961822 . OpenUrl CrossRef PubMed 43. ↵ Chen C , Xie T , Zhang Y , Wang Y , Yu F , Lin L , Zhang W , Brown BC , Zhang X , Kellems RE , D’Alessandro A , Xia Y. Erythrocyte ENT1-AMPD3 Axis is an Essential Purinergic Hypoxia Sensor and Energy Regulator Combating CKD in a Mouse Model . J Am Soc Nephrol . 2023 Oct 1 ; 34 ( 10 ): 1647 - 1671 . eng. A. D’Alessandro reports Consultancy: Hemanext Inc., Macopharma Inc.; and Ownership Interest: Omix Technologies Inc. All remaining authors have nothing to disclose. Epub 20230807. doi: 10.1681/asn.0000000000000195 . Cited in: Pubmed; PMID 37725437 . OpenUrl CrossRef PubMed 44. ↵ D’Alessandro A , Yoshida T , Nestheide S , Nemkov T , Stocker S , Stefanoni D , Mohmoud F , Rugg N , Dunham A , Cancelas JA . Hypoxic storage of red blood cells improves metabolism and post-transfusion recovery . Transfusion . 2020 Apr ; 60 ( 4 ): 786 – 798 . Epub 2020/02/28. doi: 10.1111/trf.15730 . Cited in: Pubmed; PMID 32104927 . OpenUrl CrossRef PubMed 45. ↵ Riddick DS , Ding X , Wolf CR , Porter TD , Pandey AV , Zhang QY , Gu J , Finn RD , Ronseaux S , McLaughlin LA , Henderson CJ , Zou L , Flück CE . NADPH-cytochrome P450 oxidoreductase: roles in physiology, pharmacology, and toxicology . Drug Metab Dispos . 2013 Jan ; 41 ( 1 ): 12 – 23 . eng. Epub 20121019. doi: 10.1124/dmd.112.048991 . Cited in: Pubmed; PMID 23086197 . OpenUrl Abstract / FREE Full Text 46. ↵ Low JJ , Tan BJ , Yi LX , Zhou ZD , Tan EK . Genetic susceptibility to caffeine intake and metabolism: a systematic review . J Transl Med . 2024 Oct 22 ; 22 ( 1 ): 961 . eng. The authors declare that they have no competing interests. Epub 20241022. doi: 10.1186/s12967-024-05737-z . Cited in: Pubmed; PMID 39438936 . OpenUrl CrossRef PubMed 47. ↵ Cornelis MC , Kacprowski T , Menni C , Gustafsson S , Pivin E , Adamski J , Artati A , Eap CB , Ehret G , Friedrich N , Ganna A , Guessous I , Homuth G , Lind L , Magnusson PK , Mangino M , Pedersen NL , Pietzner M , Suhre K , Völzke H , team SKPoGiH , Bochud M , Spector TD , Grabe HJ , Ingelsson E . Genome-wide association study of caffeine metabolites provides new insights to caffeine metabolism and dietary caffeine-consumption behavior . Human Molecular Genetics . 2016 ; 25 ( 24 ): 5472 – 5482 . doi: 10.1093/hmg/ddw334 . OpenUrl CrossRef PubMed 48. ↵ Nemkov T , Skinner SC , Nader E , Stefanoni D , Robert M , Cendali F , Stauffer E , Cibiel A , Boisson C , Connes P , D’Alessandro A . Acute Cycling Exercise Induces Changes in Red Blood Cell Deformability and Membrane Lipid Remodeling . Int J Mol Sci . 2021 Jan 18 ; 22 (2). eng. Epub 2021/01/23. doi: 10.3390/ijms22020896 . Cited in: Pubmed; PMID 33477427 . OpenUrl CrossRef PubMed 49. ↵ Nemkov T , Stauffer E , Cendali F , Stephenson D , Nader E , Robert M , Skinner S , Dzieciatkowska M , Hansen KC , Robach P , Millet G , Connes P , D’Alessandro A . Long-Distance Trail Running Induces Inflammatory-Associated Protein, Lipid, and Purine Oxidation in Red Blood Cells . bioRxiv . 2025 :2025.04.09.648006. doi: 10.1101/2025.04.09.648006 . OpenUrl Abstract / FREE Full Text View the discussion thread. Back to top Previous Next Posted May 30, 2025. Download PDF Supplementary Material Email Thank you for your interest in spreading the word about bioRxiv. NOTE: Your email address is requested solely to identify you as the sender of this article. Your Email * Your Name * Send To * Enter multiple addresses on separate lines or separate them with commas. You are going to email the following Caffeine Impairs Red Blood Cell Storage Quality by Dual Inhibition of ADORA2b Signaling and G6PD Activity Message Subject (Your Name) has forwarded a page to you from bioRxiv Message Body (Your Name) thought you would like to see this page from the bioRxiv website. Your Personal Message CAPTCHA This question is for testing whether or not you are a human visitor and to prevent automated spam submissions. Share Caffeine Impairs Red Blood Cell Storage Quality by Dual Inhibition of ADORA2b Signaling and G6PD Activity Monika Dzieciatkowska , Ariel Hay , Aaron Issaian , Gregory R. Keele , Shaun Bevers , Travis Nemkov , Julie A Reisz , Mark Maslanka , Daniel Stephenson , Amy L Moore , Xutao Deng , Mars Stone , Kirk C. Hansen , Steve Kleinman , Philip J Norris , Michael P. Busch , Grier P Page , Nareg Roubinian , Yang Xia , James C Zimring , Angelo D’Alessandro bioRxiv 2025.05.27.656446; doi: https://doi.org/10.1101/2025.05.27.656446 Share This Article: Copy Citation Tools Caffeine Impairs Red Blood Cell Storage Quality by Dual Inhibition of ADORA2b Signaling and G6PD Activity Monika Dzieciatkowska , Ariel Hay , Aaron Issaian , Gregory R. Keele , Shaun Bevers , Travis Nemkov , Julie A Reisz , Mark Maslanka , Daniel Stephenson , Amy L Moore , Xutao Deng , Mars Stone , Kirk C. Hansen , Steve Kleinman , Philip J Norris , Michael P. Busch , Grier P Page , Nareg Roubinian , Yang Xia , James C Zimring , Angelo D’Alessandro bioRxiv 2025.05.27.656446; doi: https://doi.org/10.1101/2025.05.27.656446 Citation Manager Formats BibTeX Bookends EasyBib EndNote (tagged) EndNote 8 (xml) Medlars Mendeley Papers RefWorks Tagged Ref Manager RIS Zotero Tweet Widget Facebook Like Google Plus One Subject Area Biochemistry Subject Areas All Articles Animal Behavior and Cognition (7636) Biochemistry (17697) Bioengineering (13896) Bioinformatics (41957) Biophysics (21457) Cancer Biology (18595) Cell Biology (25522) Clinical Trials (138) Developmental Biology (13381) Ecology (19903) Epidemiology (2067) Evolutionary Biology (24323) Genetics (15612) Genomics (22511) Immunology (17738) Microbiology (40401) Molecular Biology (17184) Neuroscience (88624) Paleontology (667) Pathology (2833) Pharmacology and Toxicology (4825) Physiology (7644) Plant Biology (15158) Scientific Communication and Education (2046) Synthetic Biology (4296) Systems Biology (9825) Zoology (2271)