Progression Of Endothelial Dysfunction Research Articles

Aging Is Associated with a Circulating Endothelial Cell-Derived Microvesicle Phenotype that Increases Cerebral Oxidative Stress

Aging is associated with increased risk and prevalence of ischemic stroke. Vascular oxidative stress impairs cell function and is considered to be an important mechanism underlying the age-related increase in cerebrovascular accidents (CVA). However, the direct, and indirect, effects of aging on endothelial cell oxidative stress are complex and not completely understood. Increase in reactive oxygen species coupled with reduced anti-oxidative capacity creates a prooxidative endothelial phenotype resulting in impaired cell dysfunction, accelerated unprogrammed cell death and, in turn, CVAs. Several recent seminal studies have demonstrated that circulating endothelial cell- derived microvesicles (EMVs) are etiologically involved in the development and progression of endothelial dysfunction and CVA. We have previously demonstrated that aging is associated with increased circulating levels of EMVs. The aim of this study was to determine, in vitro, the effect of circulating EMVs isolated from young and older adults on brain endothelial cell oxidative stress. We hypothesized that EMVs from older adults would induce a prooxidative endothelial phenotype. As part of an ongoing study, 16, non-obese, normotensive, sedentary adults were studied: 8 young (5M/3F; age: 25±1 yr) and 8 older (4M/4F; 69±2 yr). EMV identification (CD144+) and isolation from peripheral blood was performed by flow cytometry. Human cerebral microvascular endothelial cells (hCMECs) were cultured and separately treated with EMVs from each subject for 24 hours. Intracellular reactive oxygen species (ROS) production was determined using CellROX Deep Red Reagent. Circulating EMVs were ~75% higher (P=0.002) in older (140±13 EMV/μL) vs young (82±8 EMV/μL) adults. Cellular ROS was significantly higher (~40%) in hCMECs treated with EMVs from older (141±14%) compared with young (101±5%) adults. In addition, intracellular expression of key anti-oxidant proteins were significantly lower in cells treated with EMVs from older adults. Cell expression of both superoxide dismutase 1 (SOD-1: 20.6±2.3 vs 36.3±3.1 AU) and catalase (17.1±1.9 vs 27.5±3.0 AU) were ~35% lower in hCMECs treated with EMVs from older adults. Moreover, intracellular ROS was inversely and significantly correlated with SOD-1 (r=-0.54; P=0.03) and catalase (r=-0.62; P=0.01) expression reflective of the cellular oxidative milieu. In conclusion, circulating EMVs may contribute to the increased risk of ischemic stroke with aging by increasing brain endothelial cell oxidative stress and reducing oxidative defense mechanisms. EMVs may represent a viable therapeutic target for mitigating the risk of CVA with aging. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Fuchs endothelial corneal dystrophy: an updated review.

The present review will summarize FECD-associated genes and pathophysiology, diagnosis, current therapeutic approaches, and future treatment perspectives. Literature review. Fuchs' endothelial corneal dystrophy (FECD) is the most common bilateral corneal dystrophy and accounts for one-third of all corneal transplants performed in the US. FECD is caused by a combination of genetic and non-heritable factors, and there are two types: early-onset FECD, which affects individuals from an early age and is usually more severe, and late-onset FECD, which is more common and typically manifests around the age of 40. The hallmark findings of FECD include progressive loss of corneal endothelial cells and the formation of focal excrescences (guttae) on the Descemet membrane. These pathophysiological changes result in progressive endothelial dysfunction, leading to a decrease in visual acuity and blindness in later stages. The present review will summarize FECD-associated genes and pathophysiology, diagnosis, current therapeutic approaches, and future treatment perspectives. With the characterization and understanding of FECD-related genes and ongoing research into regenerative therapies for corneal endothelium, we can hope to see more significant improvements in the future in the management and care of the disease.

The effect of anthracycline-based chemotherapy on endothelial function and arterial stiffness in breast cancer patients

Abstract Introduction Cardiotoxicity restricts anthracycline and trastuzumab treatment of Human Epidermal Growth Factor Receptor 2 positive early breast cancer. Endothelial dysfunction and arteriosclerosis are significant cardiovascular risk factors. Purpose We studied the effect of anthracycline-based chemotherapy, with or without trastuzumab, on endothelium and arteriosclerosis in patients with breast cancer. Methods In this case-control study, 52 women with breast cancer and 104 women without breast cancer were examined longitudinally up to 15 months following (in the breast cancer group) initiation of chemotherapy. Arterial stiffness was evaluated through pulse wave velocity (PWV), while endothelial function via flow-mediated dilatation (FMD) at baseline (T0), 3 (T1), 6 (T2), and 15 (T3) months later. Results There was no difference between subjects with breast cancer and control in PWV and FMD at baseline. Longitudinally, participants with breast cancer exhibited considerable impairment of PWV and FMD compared to the control group (p for interaction < 0.001 for both parameters). In breast cancer patient there was a significant increase from T0 to T3 in PWV (7.43 ± 1.68 m/sec vs. 8.18 ± 2.00 m/sec, p = 0.01) and decrease in FMD (6.95 ± 2.86 % vs. 5.03 ± 2.83 %, p = 0.006). The addition of trastuzumab in the treatment did not have any effect on PWV (p = 0.74) or FMD (p = 0.91). Conclusions In patients with breast cancer, there is progression of endothelial dysfunction and arteriosclerosis up to 15 months following initiation of anthracycline-based chemotherapy. Trastuzumab has no additive effect on endothelial function or arterial stiffness.Figure 1Figure 2

RISK OF DEVELOPMENT CARDIOVASCULAR СOMPLICATIONS IN PATIENTS WITH ARTERIAL HYPERTENSION AND METABOLIC SYNDROME

The combination of arterial hypertension and metabolic syndrome leads to an increased risk of development cardiovascular complications. Early detection of asymptomatic target organ damage to determine additional risk is important, because pressure-dependent subclinical damage to several organs reflects progression along the cardiovascular disease continuum. The SCORE scale is based on data from a number of recent European multicenter studies and takes into account all variants of fatal cardiovascular events over a 10-year period. However, the SCORE scale does not take into account the presence of such subclinical risk factors for the development of cardiovascular complications as insulin resistance, endothelial dysfunction, microalbuminuria
 The aim was to assess the ten-years risk of development of cardiovascular complications in hypertensive's with metabolic syndrome, depending on the type of insulinemia, endothelial dysfunction, and the presence of microalbuminuria.
 Materials and methods. Were examined 96 patients (34 men, 62 women) with arterial hypertension and metabolic syndrome, average age (57±9) years. Office blood pressure measurement, the level of total cholesterol in the blood and microalbuminuria in daily urine were determined for all patients. An oral glucose tolerance test was performed with parallel determination of the level of endogenous insulin, endothelin-1, and vasculoendothelial growth factor in blood serum by the immunoenzymatic method. Patients were divided into three groups. Group 1 included 29 patients with normal basal and postprandial levels of endogenous insulin; in the 2nd group – 39 patients with reactive hyperinsulinemia, in the 3rd group – 28 patients with spontaneous hyperinsulinemia. To estimate the total cardiovascular risk, the version of the SCORE scale for countries with high cardiovascular mortality was used. Moderate risk criteria are defined – SCORE index level 1-4%, high risk – 5-9% and very high risk – ≥ 10%.
 Research results. A moderate cardiovascular risk prevailed in almost 52% of patients of group 1 with a normal level of endogenous insulin, high – in 35% of patients group 2 with reactive hyperinsulinemia, very high – in 48% of patients group 2 and 43% of patients of group 3 with spontaneous hyperinsulinemia. The highest levels of systolic and diastolic blood pressure were characteristic of high-risk patients with spontaneous hyperinsulinemia combined with impaired glucose tolerance, with increasing of total cholesterol levels in 1.7 times under obesity, increasing of endothelin levels by 7 times, vasculoendothelial growth factor in 2.7 times and the presence of microalbuminuria in 75% of cases.
 Conclusions. More than half patients with arterial hypertensive and metabolic syndrome have a high and very high cardiovascular risk according to the SCORE scale, which is associated with hyperinsulinemia and insulin resistance, hypercholesterolemia and obesity, microalbuminuria and disorders of endothelial vasoregulation. The risk of cardiovascular complications based on subclinical signs of target organs damage to increases with the severity of insulin resistance and progression of endothelial dysfunction, which creates prerequisites for the development of atherosclerosis.

Prolonged impact of anti-cancer therapy on endothelial function and arterial stiffness in breast cancer patients

BackgroundCardiotoxicity restricts anthracycline and trastuzumab treatment of Human Epidermal Growth Factor Receptor 2 positive early breast cancer. Endothelial dysfunction and arteriosclerosis are significant cardiovascular risk factors. ObjectivesWe studied the effect of anthracycline-based chemotherapy, with or without trastuzumab, on endothelium and arteriosclerosis in patients with breast cancer. MethodsIn this case-control study, 52 women with breast cancer and 104 women without breast cancer were examined longitudinally up to 15 months following (in the breast cancer group) initiation of chemotherapy. Arterial stiffness was evaluated through pulse wave velocity (PWV), while endothelial function via flow-mediated dilatation (FMD) at baseline (T0), 3 (T1), 6 (T2), and 15 (T3) months later. ResultsThere was no difference between subjects with breast cancer and control in PWV and FMD at baseline. Longitudinally, participants with breast cancer exhibited considerable impairment of PWV and FMD compared to the control group (p for interaction <0.001 for both parameters). In breast cancer patients, there was a significant increase from T0 to T3 in PWV (7.43 ± 1.68 m/s vs. 8.18 ± 2.00 m/s, p = 0.01) and decrease in FMD (6.95 ± 2.86% vs. 5.03 ± 2.83%, p = 0.006). The addition of trastuzumab in the treatment did not have any effect on PWV (p = 0.74) or FMD (p = 0.91). ConclusionsIn patients with breast cancer, there is progression of endothelial dysfunction and arteriosclerosis up to 15 months following initiation of anthracycline-based chemotherapy. Trastuzumab has no additive effect on endothelial function or arterial stiffness.

Chronic anemia is associated with systemic endothelial dysfunction.

In acute myocardial infarction and heart failure, anemia is associated with adverse clinical outcomes. Endothelial dysfunction (ED) is characterized by attenuated nitric oxide (NO)-mediated relaxation responses which is poorly studied in chronic anemia (CA). We hypothesized that CA is associated with ED due to increased oxidative stress in the endothelium. CA was induced by repeated blood withdrawal in male C57BL/6J mice. Flow-Mediated Dilation (FMD) responses were assessed in CA mice using ultrasound-guided femoral transient ischemia model. Tissue organ bath was used to assess vascular responsiveness of aortic rings from CA mice, and in aortic rings incubated with red blood cells (RBCs) from anemic patients. In the aortic rings from anemic mice, the role of arginases was assessed using either an arginase inhibitor (Nor-NOHA) or genetic ablation of arginase 1 in the endothelium. Inflammatory changes in plasma of CA mice were examined by ELISA. Expression of endothelial NO synthase (eNOS), inducible NO synthase (iNOS), myeloperoxidase (MPO), 3-Nitrotyrosine levels, and 4-Hydroxynonenal (4-HNE) were assessed either by Western blotting or immunohistochemistry. The role of reactive oxygen species (ROS) in ED was assessed in the anemic mice either supplemented with N-Acetyl cysteine (NAC) or by in vitro pharmacological inhibition of MPO. The FMD responses were diminished with a correlation to the duration of anemia. Aortic rings from CA mice showed reduced NO-dependent relaxation compared to non-anemic mice. RBCs from anemic patients attenuated NO-dependent relaxation responses in murine aortic rings compared to non-anemic controls. CA results in increased plasma VCAM-1, ICAM-1 levels, and an increased iNOS expression in aortic vascular smooth muscle cells. Arginases inhibition or arginase1 deletion did not improve ED in anemic mice. Increased expression of MPO and 4-HNE observed in endothelial cells of aortic sections from CA mice. NAC supplementation or inhibition of MPO improved relaxation responses in CA mice. Chronic anemia is associated with progressive endothelial dysfunction evidenced by activation of the endothelium mediated by systemic inflammation, increased iNOS activity, and ROS production in the arterial wall. ROS scavenger (NAC) supplementation or MPO inhibition are potential therapeutic options to reverse the devastating endothelial dysfunction in chronic anemia.

Aging Is Associated with a Circulating Endothelial Cell-Derived Microvesicle Phenotype that Impairs Cerebral Fibrinolytic Capacity

Aging is associated with increased risk and prevalence of ischemic stroke. Endothelial cell dysfunction is considered to be a central mechanism underlying the age-related increase in cerebrovascular accidents (CVA). However, the direct, and indirect, effects of aging on endothelial cell biology are diverse and not completely understood. Reduced endogenous fibrinolytic activity is a central etiologic factor underlying ischemic stroke. Endothelial cells are the primary site of synthesis and release of tissue-type plasminogen activator (t-PA), the main plasminogen activator in fibrinolysis and central regulator. Clinical interest in circulating extracellular vesicles, particularly endothelial cell-derived microvesicles (EMVs), has intensified due to their involvement in the development and progression of endothelial dysfunction and CVA. The aim of this study was to determine, in vitro, the effect of circulating EMVs isolated from young and older adults on brain endothelial cell fibrinolytic capacity. We hypothesized that EMVs from older adults would reduce brain endothelial cell t-PA production and release and increase PAI-1 production compared with EMVs from young adults. Twenty-two healthy, non-obese, normotensive, sedentary adults were studied: 10 young (6M/4F; age: 25±1 yr) and 12 older (7M/5F; 64±2 yr). EMV identification (CD144 + ) and isolation from peripheral blood were performed by flow cytometry. Human cerebral microvascular endothelial cells (hCMECs) were cultured and separately treated with EMVs from each subject. Cultured hCMECs were treated with EMVs in the absence and presence of thrombin (1 unit/mL) for 24 hours. Basal expression of t-PA (25.2±1.5 vs 39.8±1.8 AU) and the t-PA response to thrombin (32.5±1.5 vs 57.2±2.2 AU) were significantly lower (~35%) in hCMECs treated with EMVs from older compared with young adults. Concordant with changes in cell protein expression, basal t-PA production (55.2±3.5 vs 70.1±3.7 ng/mL) and thrombin-stimulated t-PA production (61.9±3.7 vs 88.4±3.8 ng/mL) was significantly lower in cells treated with EMVs from older adults. In conclusion, circulating EMVs may contribute to the increased risk of ischemic stroke with aging by impairing brain endothelial cell fibrinolytic capacity. Diminished capacity of the endothelium to release t-PA is a key mechanistic factor underlying cerebral thrombosis. Nothing to Disclose This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Interrelation between the parameters of endogenous vascular regulation, oxidative stress and the markers of inflammatory response in COVID-19 patients while on extracorporeal membrane oxygenation

Introduction. Extracorporeal membrane oxygenation has found wide application in clinical practice during the COVID-19 pandemic. Oxidative stress, endothelial dysfunction, and systemic inflammatory response syndrome play an important role in the pathogenesis of COVID-19. Our research was designed to study correlations in-between those factors and the impact of extracorporeal membrane oxygenation on them.Aim. The study of systemic inflammatory response and endothelial function in patients with COVID-19 during extracorporeal membrane oxygenation.Material and methods. In the course of a prospective study, we examined 100 COVID-19 patients aged 26 to 75 years, median 55 years [47;60], who were treated at the N.V. Sklifosovsky Research Institute for Emergency Medicine, using extracorporeal membrane oxygenation. As a control group (normal), 25 practically healthy people whose median age was 32 years [25;39] were examined. The function of the vascular endothelium was assessed by the content of nitric oxide stable metabolites in the blood serum and the level of angiotensin-converting enzyme. Next, the ratio of nitric oxide metabolite to angiotensin-converting enzyme level was calculated, reflecting the imbalance between endotheliumdependent vasodilation and vasoconstriction. To assess the severity of oxidative stress in blood serum, malondialdehyde was determined as a marker of lipid peroxidation. The state of the antioxidant system was assessed in terms of total antioxidant status of blood serum. The presence of an imbalance in the system of lipid peroxidation and the antioxidant system total antioxidant status was judged by the oxidative stress coefficient, i.e. the ratio of malondialdehyde to the total antioxidant activity.Results. The analysis showed the presence and progression of endothelial dysfunction, impaired vascular regulation, activation of free radical processes, the presence of an imbalance in the prooxidant/antioxidant system, as well as the progression of the inflammatory process with a decrease in the level of markers of the COVID-19 severity.Conclusion. Further studies of the correlation between endothelial damage and the severity of the systemic inflammatory response syndrome may be of fundamental importance for explaining the pathophysiological mechanisms of COVID-19 course and developing new treatments for such patients.

Coronary heart disease in patients with human immunodeficiency virus infection

According to the World Health Organization, more than 38 million people are carriers of the human immunodeficiency virus (HIV). The relative risk of cardiovascular events in patients with HIV infection is 1.52 times higher than those without it. An HIV-positive status is associated with a 50% increased risk of acute myocardial infarction. The mechanisms of CVD development caused by HIV activity are not fully understood, and the contribution of traditional risk factors cannot fully explain such a high potential for coronary heart disease (CHD) in patients with HIV infection. Data suggest that HIV-associated chronic inflammation and immune system activation may lead to progressive endothelial dysfunction and vascular damage. The pathogenesis of atherosclerosis and, consequently, CHD in HIV infection is extremely versatile and poorly studied. The key mechanisms in HIV-associated CHD include the effect of HIV proteins on immunocompetent cells and vascular endothelium, immunodeficiency, translocation of microorganisms from the gut, and chronic inflammation with platelet activation. Despite considerable progress in understanding HIV-associated cardiovascular disease, many unresolved questions remain. Even with effective viral suppression, inflammation and immune dysregulation appear to increase the risk of cardiovascular complications. The lack of large-scale clinical studies on the prevention and treatment of cardiovascular disease in HIV is very important. Nevertheless, some important features should be considered in the management of patients with HIV infection.

Biphasic Neutrophil Recovery after CD19 CART in R/R LBCL Is Associated with Superior PFS/OS, Robust CAR T-Cell Expansion in Relation to Baseline Tumor Volume, and a Decrease of Systemic Inflammation over Time

Background: Hematological toxicity and concomitant infectious complications drive the toxicity burden of CD19 CART. We recently proposed a classification system for CART-related hematotoxicity, defining three unique phenotypes of hematopoietic recovery (Rejeski et al, Blood 2021). The underlying mechanisms of these phenotypes and their relation to survival remain poorly understood. Here, we sought to understand patterns of post-CART neutrophil recovery and their influence on clinical outcomes. Methods: In this multicenter retrospective study, we analyzed 344 pt. receiving standard-of-care axi-cel (n=208) or tisa-cel (n=136) for R/R LBCL. The phenotypes of neutrophil recovery were defined as follows: Quick: sustained neutrophil recovery without a second dip below an ANC<1000/µl.Intermittent: neutrophil recovery (ANC>1500/µl) followed by a second dip with an ANC<1000/µl after day 21.Aplastic: continuous severe neutropenia (ANC<500/µl) ≥14 days. Multivariate analysis of patient-, disease-, and therapy-related factors was performed as binary logistic regression for the aplastic vs. non-aplastic phenotypes. Variables with a p<0.2 on univariate analysis were included. Clinical outcomes were studied via Kaplan-Meier estimates. Quantification of CAR T-cells was performed by qPCR or by flow cytometry in 146 pt. according to institutional protocols. To harmonize CAR T-cell expansion across centers, patients were divided into quartiles by their respective peak expansion. Plasma proteomics were performed in 56 pts. across four time points (day 0, 4, 14, 28) using a 92-protein multiplex proximity extension assay (Olink Proteomics). Key candidate serum markers were validated in 70 pt. using the Ella automated immunoassay system (ProteinSimple). Results: Across all patients, the distribution of the 'aplastic' (A), 'intermittent' (I) and 'quick' (Q) phenotypes was 18%, 42%, and 40%, respectively. The 'aplastic' patients exhibited a longer median duration of severe neutropenia, (A vs. I vs. Q: 26 vs. 10 vs. 5 days, p<0.001), a higher rate of severe infections (A vs. I vs. Q: 41% vs. 23% vs. 18%) and a trend towards higher 1-year non-relapse mortality (A vs. I: 9% vs. 3.5%, p=0.08). On multivariate regression (n=344), the aplastic phenotype was independently associated with the presence of BM infiltration (adjusted OR=3.2, p=0.004), ANC (aOR=0.28, p=0.002), and ferritin (aOR=2.3, p=0.039). Neither high-grade CRS or ICANS nor CAR product were linked to the aplastic phenotype. In the 'intermittent' cohort, we noted a significant decrease of both serum ferritin (median 776 vs. 321 U/L) and CRP levels (median 1.6 vs. 0.2 mg/dL) between CAR infusion and day 90 follow-up. When studying CD3+ CAR T-cell dynamics over time, we found that patients with 'intermittent' recovery displayed both an earlier and more pronounced expansion peak, and longer CART persistence compared to their 'quick' counterparts. While the 'aplastic' cohort displayed comparable peak CART expansion, these patients also exhibited high baseline LDH, indicative of a higher baseline tumor burden, or 'hill to climb' (top right quadrant, Fig. 1A). Conversely, the 'quick' cohort was characterized by both low expansion and low LDH. Serum proteome analysis revealed significant disruptions of endothelial function and immune dysregulation in the 'aplastic' patients at month 1 (Fig. 1B). Importantly, we confirmed higher peak IFNγ and IL-15 levels, and an increased ratio of ANG2 to ANG1 in the 'aplastic' patients in the independent validation cohort. Notably, progression-free (PFS) and overall survival (OS) was poor in the 'aplastic' patients (1-yr PFS 26%, 1-yr OS 46%). On the other hand, 'intermittent' patients exhibited the best survival outcomes (HRPFS I vs. A: 0.45, 95% CI 0.31-0.65). Median PFS was 3 vs. 20 vs. 4 months (A vs. I vs. Q, p<0.001); median OS was 8 months vs. not-reached vs. 23 months (A vs. I vs. Q, p<0.001). Conclusion: The 'aplastic' phenotype is driven by baseline inflammation and hematopoietic function, characterized by progressive endothelial dysfunction, and associated with poor PFS/OS. In contrast, the 'intermittent' phenotype (i.e. biphasic cytopenia) was associated with high levels of CART expansion and persistence, a decrease in systemic inflammation over time, and excellent survival outcomes. Overall, these data suggest potential interactions between host hematopoiesis and CART function and efficacy. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Distinguishing Early Infections from CRS with Routine and Exploratory Serum Proteomics and the HT10 Score Following CD19 CAR-T for Relapsed/Refractory B-NHL

Background: Early fever after CAR-T therapy can reflect both an infection or CRS. Distinguishing both etiologies in these commonly neutropenic patients represents an unresolved clinical challenge. Accordingly, the vast majority of patients receive empiric broad-spectrum antibiotics, which can promote antibiotic resistance and negatively influence the gut microbiome, which has recently been linked to inferior CAR-T outcomes (Smith et al, Nature Medicine 2022). Here, we aimed to characterize infection- vs. CRS-specific differences in routine and exploratory serum proteomics, and to develop a clinical tool that reliably discriminates between both. Methods: Early fever events (day 0-30) were retrospectively characterized as CRS vs. infection in 62 patients treated with standard-of-care CD19 CAR-T for r/r B-NHL (23 Axi-cel, 30 Tisa-cel, 9 Brexu-cel). Severe infections were defined as requiring IV anti-infectives and/or hospitalization in the presence of microbiologic evidence or distinct clinical signs of infection; CRS was graded according to ASTCT criteria. Cumulative incidence curves were calculated as time-to-event-analysis for CRS 1°, ≥2° and severe infections (day of event = index event). Serum inflammatory markers (CRP, interleukin-6 [IL-6], procalcitonin [PCT]) were measured daily from CAR infusion until discharge and on subsequent outpatient visits. Inflammatory markers were tested using binary logistic regression for severe infection vs. CRSonlyindex events. Discriminatory thresholds were determined from receiver operating characteristic (ROC) curves by optimizing the Youden J statistic. Longitudinal plasma proteomics was performed in 52 patients across four time points (day 0, 4, 14, 28) using a 92-protein multiplex proximity extension assay (Olink Proteomics). We applied a linear mixed model (LMM) accounting for both infection vs. CRS status and time effects. Results: The cumulative 30-day incidence of CRS 1°, CRS ≥2° and severe infections was 47%, 43%, and 25%, respectively. Five patients had neither CRS nor an infection, and were subsequently excluded from further analysis. Tocilizumab was applied in the majority of CRS patients (1°: 90%, ≥2°: 100%). We did not find a significant correlation between CRS grade and severe infections (G2 = 0.13, p = n.s.). Median onset of CRS and infection was 3 and 8 days, respectively. Most events (54%) occurred in the setting of coincident severe neutropenia (ANC <500/µl). While event-day CRP was comparable between infection and CRS (median 1.8 vs. 3.4 mg/dL, n.s.), we noted increased serum IL-6 (median 2243 vs. 64 pg/mL, p=0.03) and particularly high PCT levels (median 1.6 vs. 0.3 µg/L, p<0.0001) in the infection cohort. For PCT, an optimal discriminatory threshold of 1.5 µg/L was established (AUCROC 0.78). In previous work, we demonstrated that a CAR-HEMATOTOX (HT) score ≥2 identifies patients with a high infection risk (Rejeski et al, JITC 2022). Indeed, the infection cohort displayed higher baseline HT scores (median 5 vs. 2, p=0.002). By incorporating day-of-fever PCT levels with the HT score, the discriminatory capacity for severe infection events was improved. On ROC analysis, the 'HT10 score' (addition of 10 points to the patient-specific HT score in case event-day PCT ≥1.5 µg/L) exhibited superior discrimination for severe infections (AUC 0.92, p<0.0001, sens. 80%, spec. 91%, Fig. 1A). Additionally, we identified protein-level differences between the infection (n=13) vs. CRSonly (n=39) cohorts from the Olink dataset. On day 0, we noted marked upregulation of the angiogenetic factor TNFRSF12A in the infection cohort (q=4.8E-05). Notably, we observed progressive endothelial dysfunction and immune dysregulation in the patients developing severe infections (day 28, Fig. 1B). This included an increased ANG2/1 ratio and dysregulated CD40/CD40L axis. Consistent with a response to microbial exposure, the innate immune regulators CXC3L1, CCL20, and CXCL13 were upregulated in the infection group. Conclusions: The high discriminatory capacity of the HT10 score for infections highlights the advantage of dynamic risk-assessment and emphasizes the utility of procalcitonin in routine inflammatory panels. If validated prospectively, the score will enable more patient-individual decisions on antibiotic use. Finally, the identified Olink candidate markers may further refine the discriminatory capacity of the HT10 score. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Peculiarities of serum levels of vascular growth factor and its receptors in dialysis patients

Recent research has shown that VEGF may influence the condition and survival of patients with chronic kidney disease (CKD), including those treated with dialysis procedures. Identifying the value of this factor is important to uncover the mechanisms of CKD progression and improve treatment strategies.&#x0D; The present study aimed to determine the mean serum levels of vascular endothelial growth factor (VEGF) and its receptor (VEGF-R) and their correlations with pro-inflammatory cytokines (TNF-α, MCP-1, IL -6, IL -8) in dialysis patients stratified by dialysis modality and Carnivit use.&#x0D; Methods. In this prospective cohort study, serum levels of VEGF, VEGF-R, and proinflammatory cytokines were determined in 41 hemodialysis (HD) patients (group 1) and 14 peritoneal dialysis (PD) patients (group 2), 28 of whom were treated with Carnivit. The assay was performed with the enzyme immunoassay "SunRise TouchScreen" using IBL International (Germany) test systems. The limits of normal values (reference range) were determined based on the results of a study with 20 conditionally healthy individuals.&#x0D; Results. All included patients had significantly higher serum levels of VEGF compared to healthy donors (p=0.006); a significant increase in this mediator was observed in both HD and PD treated subjects (p=0.002 and p=0.007, respectively). Carnivit treatment resulted in a significant decrease (p=0.023) in this mediator to normal levels in all patients. Subgroup analysis showed a significant decrease in VEGF (p=0.016) and VEGF-R (p=0.005) in HD patients after therapy. Mean concentrations of growth factor in HD patients (p=0.098) and PD patients (p=0.160) did not differ from those of healthy subjects.&#x0D; A positive correlation was observed between serum levels of VEGF and its receptors in the studied patients both before and after treatment (p &lt; 0.0001).&#x0D; A direct correlation between IL -8 and VEGF and VEGF-R was observed in HD patients and an inverse correlation between MCP-1 and VEGF-R was observed in PD patients. After Carnivit treatment, the correlation between serum levels of IL -8 and VEGF-R (0.782, p=0.01) and between VEGF and VEGF-R (0.770, p=0.01) remained in group 1 and between VEGF and VEGF-R (0.829, p=0.03) in group 2.&#x0D; Conclusion. Increased concentrations of VEGF and its receptors were detected in the blood of dialysis patients, with a positive correlation between them and the pro-inflammatory IL -8. Treatment with Carnivit resulted in a significant decrease in cytokines with the maintenance of the balance between vascular growth factor and its receptors, which we consider a positive effect for the inhibition of inflammation with the progression of endothelial dysfunction.

Descemet Stripping Endothelial Keratoplasty for Congenital Aniridia: An Interesting and Challenging Story

Congenital aniridia is a rare condition affecting a wide range of ocular structures, from the ocular surface to the retina. We present the case of a 59-year-old woman with PAX6- and WT1-negative congenital aniridia who developed aniridia-associated keratopathy and progressive endothelial dysfunction with corneal decompensation after cataract surgery. The patient underwent successful ultrathin Descemet stripping endothelial keratoplasty. Despite the challenges faced with an unstable iridolenticular diaphragm, we were pleasantly surprised to see improvement not only of corneal edema and endothelial function but also of the whole cornea, including anterior corneal anatomy and appearance. In conclusion, endothelial transplantation in a patient with aniridia resulted in improvement of all the corneal structures from the endothelium to the stroma, epithelium, and possibly even the ocular surface.

P1458: PATTERNS OF LONG-TERM HEMATOPOIETIC RECOVERY IMPACT SURVIVAL OUTCOMES AFTER CD19-DIRECTED CAR T-CELL THERAPY FOR R/R LBCL

Background: Real-world evidence has underlined the role of hematological toxicity and infections in driving the toxicity burden of CD19 CAR-T. We have recently developed a classification system for post-CAR-T hematotoxicity, identifying three unique phenotypes of neutrophil recovery (Rejeski et al Blood 2021). However, their prognostic impact remains poorly understood. Aims: We sought to understand potential pathomechanisms underlying hematopoietic recovery patterns, and characterize their influence on toxicity and clinical outcomes. Methods: In this multicenter retrospective observational study, we analyzed 316 pts. receiving Axicabtagene ciloleucel (n=194) or Tisagenlecleucel (n=122) for R/R LBCL in a real-world setting. The phenotypes of neutrophil recovery were defined as follows: 1) Quick Recovery: sustained neutrophil recovery without a second dip below an ANC<1000/µl. 2) Intermittent Recovery: neutrophil recovery (ANC>1500/µl) followed by a second dip with an ANC<1000/µl after day 21. 3) Aplastic: continuous severe neutropenia (ANC<500/µl) ≥14 days. Multivariate analysis was performed as stepwise binary logistic regression for the aplastic vs. non-aplastic phenotypes. Variables with a p<0.2 on univariate analysis were included. Clinical outcomes were studied via Kaplan-Meier estimates. Serum cytokines were analyzed using the Ella automated immunoassay system (ProteinSimple) in 70 pt. from Moffitt Cancer Center. The plasma proteome was characterized in a further 38 pts. from the LMU Munich across four time points (day 0, 4, 14, 28) using a 92-protein multiplex proximity extension assay (Olink Bioscience). Results: The overall distribution of the ‘quick’ (Q), ‘intermittent’ (I) and ‘aplastic’ (A) phenotypes was 40%, 41%, and 19%, respectively. Prior to lymphodepletion, ‘aplastic’ patients displayed increased systemic inflammation (median ferritin 1168 ng/ml), impaired hematopoietic function (median platelet count 101 G/l) and high tumor burden (median LDH 344 U/l). ‘Aplastic’ patients displayed a longer median duration of severe neutropenia, (A vs. I vs. Q: 28 vs. 10 vs. 5 days, p<0.001), higher rate of severe infections (A vs. I vs. Q: 40% vs. 22% vs. 18%), and a trend towards higher 1-year non-relapse mortality (A vs. I: 10% vs. 4%, p=0.09). Progression-free (PFS) and overall survival (OS) was poor in the ‘aplastic’ patients (Fig. 1, 1-yr PFS 24%, 1-yr OS 48%). On the other hand, ‘intermittent’ patients exhibited the best survival outcomes (HR for PFS, I vs. A: 0.46, 95% CI 0.32-0.69). Median PFS was 3 vs. 4 vs. 20 months (A vs. Q v. I, p<0.001); median OS was 8 vs. 23 months vs. not reached (A vs. Q v. I, p=0.0025). On multivariate regression (n=316), the aplastic phenotype was primarily driven by baseline ECOG (OR=1.6, p=0.05), ferritin (OR=2.5, p=0.01), ANC (OR=0.3, p=0.01), and hemoglobin (OR=0.8, p=0.04). Of note, high-grade CRS/ICANS and CAR product were not associated with the aplastic phenotype. Initial analyses of inflammatory patterns revealed higher peak IL-15 levels and an increased ratio of ANG2 to ANG1 in ‘aplastic’ patients in both cohorts, with ANG-1 levels decreasing over time. Image:Summary/Conclusion: Hematopoietic recovery patterns impact disease outcomes after CD19 CAR-T. Surprisingly, a phenotype associated with recurrent neutrophil dips (i.e. prolonged cytopenia) was associated with improved survival outcomes. The poor-risk ‘aplastic’ phenotype was driven by baseline inflammation and hematopoietic function, and characterized by progressive endothelial dysfunction. Further mechanistic exploration is warranted and ongoing.

GESTATIONAL DIABETES AS A POSSIBLE PREDICTOR OF LATE PREECLAMPSIA

Abstract. Purpose of the study - We assessed the impact of gestational diabetes on the development of preeсlampsia. Material and methods of research: We conducted a retrospective case-control study. Group I included 34 patients with preeclampsia, group II included 66 patients without preeclampsia. We analyzed the course of pregnancy and the development of gestational diabetes. Research results. In the group of patients with late preeclampsia (after 34 weeks of pregnancy), we found a high incidence of gestational diabetes mellitus. The frequency of gestational diabetes was noted in 56% of cases, while in the group of healthy pregnant women, the incidence of this pathology is 4.55% as the average in the population. Conclusions. Gestational diabetes mellitus can be considered one of the predictors of late preeclampsia. The course of diabetes is associated with progressive endothelial dysfunction, which can aggravate the existing compensated placental ischemia, which is one of the important pathogenetic links in the implementation of preeclampsia. We recommend explaining not only the risks of the underlying disease, but also careful self-monitoring of blood pressure at home in patients with diabetes. We also recommend testing for preeclampsia after detection of gestational diabetes mellitus (placental growth factor/fms-like soluble tyrosine kinase-1), which provides information on the likely debut of a hypertensive disorder within 4 weeks ahead.

Personalized management of hypertensive patients: focus on prognostic biomarkers

Aim. To analyze level of circulating biomarkers of plasma vascular endothelial growth factor (VEGF) and tumor necrosis factor-α (TNF-α) during 3-month therapy with a fixed-dose combination of ramipril/indapamide (Konsilar-D24, AO Vertex, Russia), as well as to evaluate the antihypertensive efficacy of a fixed-dose combination of ramipril/indapamide (Konsilar-D24, JSC “Vertex”, Russia) in hypertensive (HTN) patients with type 2 diabetes (T2D).Material and methods. This prospective open-label study included 44 patients (aged 35 to 60 years) of both sexes with essential grade 1-2 HTN and concomitant compensated T2D, who did not reach the target blood pressure (BP) level using single or dual antihypertensive therapy, as well as patients who did not take antihypertensives. All patients included in the study initially underwent a set of standard clinical, laboratory and functional examinations in accordance with the clinical guidelines for the management of patients with HTN and T2D, as well as an assessment of the level of C-reactive protein, VEGF and TNF-α. Patients were monitored and treated with Konsilar-D24 for 3 months.Results. In 93,2% of patients, individual target BP values were achieved during the first 2-4 weeks of therapy with a fixed combination of ramipril/indapamide (Konsilar-D24). In the subsequent 3-month follow-up, the average daily BP level in all patients ranged from 129/79 mm Hg to 110/70 mm Hg. Three-month Konsilar-D24 therapy showed a decrease in microalbuminuria: the median values of microalbuminuria decreased by 2 times, and the decrease in the maximum recorded values reached 40% of the baseline. Decrease in mean TNF-α values after 3-month therapy with Konsilar-D24 was 33% of the baseline values, while the maximum recorded values during the specified period decreased by 17%. Decrease in median VEGF values after 3-month Konsilar-D24 therapy was 28%, while the maximum value decreased by 7%, the minimum — by 8%.Conclusion. Konsilar-D24 improves the prognosis in hypertensive patients not only by reducing BP to target values, but also by reducing the level of VEGF and TNF-α biomarkers that determine the progression of endothelial dysfunction, diabetic retinopathy, and microalbuminuria.

Personalized management of hypertensive patients: focus on prognostic biomarkers

Aim. To analyze level of circulating biomarkers of plasma vascular endothelial growth factor (VEGF) and tumor necrosis factor-α (TNF-α) during 3-month therapy with a fixed-dose combination of ramipril/indapamide (Konsilar-D24, AO Vertex, Russia), as well as to evaluate the antihypertensive efficacy of a fixed-dose combination of ramipril/indapamide (Konsilar-D24, JSC “Vertex”, Russia) in hypertensive (HTN) patients with type 2 diabetes (T2D).Material and methods. This prospective open-label study included 44 patients (aged 35 to 60 years) of both sexes with essential grade 1-2 HTN and concomitant compensated T2D, who did not reach the target blood pressure (BP) level using single or dual antihypertensive therapy, as well as patients who did not take antihypertensives. All patients included in the study initially underwent a set of standard clinical, laboratory and functional examinations in accordance with the clinical guidelines for the management of patients with HTN and T2D, as well as an assessment of the level of C-reactive protein, VEGF and TNF-α. Patients were monitored and treated with Konsilar-D24 for 3 months.Results. In 93,2% of patients, individual target BP values were achieved during the first 2-4 weeks of therapy with a fixed combination of ramipril/indapamide (Konsilar-D24). In the subsequent 3-month follow-up, the average daily BP level in all patients ranged from 129/79 mm Hg to 110/70 mm Hg. Three-month Konsilar-D24 therapy showed a decrease in microalbuminuria: the median values of microalbuminuria decreased by 2 times, and the decrease in the maximum recorded values reached 40% of the baseline. Decrease in mean TNF-α values after 3-month therapy with Konsilar-D24 was 33% of the baseline values, while the maximum recorded values during the specified period decreased by 17%. Decrease in median VEGF values after 3-month Konsilar-D24 therapy was 28%, while the maximum value decreased by 7%, the minimum — by 8%.Conclusion. Konsilar-D24 improves the prognosis in hypertensive patients not only by reducing BP to target values, but also by reducing the level of VEGF and TNF-α biomarkers that determine the progression of endothelial dysfunction, diabetic retinopathy, and microalbuminuria.

The use of microvascular endothelial function indicators to calculate functional age of the microcirculation system

Aim. To find out the possibility of using microvascular endothelial function indicators to determine the biological age (BA) of a person. Materials and methods. 390 patients aged 20–29 years (n = 31), 30–39 (n = 22), 40–49 (n = 45), 50–59 years (n = 72), 60–69 years (n = 129) and 70–79 years (n = 91) were examined. The functional state of the microvascular endothelium was analyzed by laser Doppler flowmetry on a BLF-21D (Transonic Systems Inc, USA) at the internal surface of the middle third of the forearm. Platelet aggregation activity was measured on a two-channel laser platelet aggregation analyzer 23 LA (Biola, Russia) by turbidimetric method, blood viscosity – using a rotational viscometer AKR-2 (Russia) at a shear rate of 10–200 s-1 with the deformation index and erythrocyte aggregation index calculation. Results. There was a statistically significant decrease in the volumetric rate of the skin blood flow (SBF) at rest and at the peak of post-occlusion reactive hyperemia, a reduced recovery time of SBF to baseline values from the age of 50–59 years, indicating endothelial dysfunction with aging. A progression of endothelial dysfunction in persons older than 60 years was accompanied by hemostatic parameters deterioration: increased blood viscosity, increased both spontaneous and induced platelet aggregation activity. Blood viscosity was statistically significantly increased from the age of 40. The formula for calculating BA using indicators of the microvascular endothelium functional state, platelet aggregation capacity and anthropometric indicators was obtained. Conclusions. The formula for calculating human BA using indicators of the microvascular endothelium functional state, platelet aggregation capacity and anthropometric indicators has been developed. Sufficiently high accuracy determination of the microcirculation system functional age based on the developed method has been shown.

Role of Ion Channel Remodeling in Endothelial Dysfunction Induced by Pulmonary Arterial Hypertension.

Endothelial dysfunction is a key player in advancing vascular pathology in pulmonary arterial hypertension (PAH), a disease essentially characterized by intense remodeling of the pulmonary vasculature, vasoconstriction, endothelial dysfunction, inflammation, oxidative stress, and thrombosis in situ. These vascular features culminate in an increase in pulmonary vascular resistance, subsequent right heart failure, and premature death. Over the past years, there has been a great development in our understanding of pulmonary endothelial biology related to the genetic and molecular mechanisms that modulate the endothelial response to direct or indirect injury and how their dysregulation can promote PAH pathogenesis. Ion channels are key regulators of vasoconstriction and proliferative/apoptotic phenotypes; however, they are poorly studied at the endothelial level. The current review will describe and categorize different expression, functions, regulation, and remodeling of endothelial ion channels (K+, Ca2+, Na+, and Cl− channels) in PAH. We will focus on the potential pathogenic role of ion channel deregulation in the onset and progression of endothelial dysfunction during the development of PAH and its potential therapeutic role.

Effects of Tolvaptan on Oxidative Stress in ADPKD: A Molecular Biological Approach.

Autosomal dominant polycystic disease (ADPKD) is the most frequent monogenic kidney disease. It causes progressive renal failure, endothelial dysfunction, and hypertension, all of which are strictly linked to oxidative stress (OxSt). Treatment with tolvaptan is known to slow the renal deterioration rate, but not all the molecular mechanisms involved in this effect are well-established. We evaluated the OxSt state in untreated ADPKD patients compared to that in tolvaptan-treated ADPKD patients and healthy subjects. OxSt was assessed in nine patients for each group in terms of mononuclear cell p22phox protein expression, NADPH oxidase key subunit, MYPT-1 phosphorylation state, marker of Rho kinase activity (Western blot) and heme oxygenase (HO)-1, induced and protective against OxSt (ELISA). p22phox protein expression was higher in untreated ADPKD patients compared to treated patients and controls: 1.42 ± 0.11 vs. 0.86 ± 0.15 d.u., p = 0.015, vs. 0.53 ± 0.11 d.u., p < 0.001, respectively. The same was observed for phosphorylated MYPT-1: 0.96 ± 0.28 vs. 0.68 ± 0.09 d.u., p = 0.013 and vs. 0.47 ± 0.13 d.u., p < 0.001, respectively, while the HO-1 expression of untreated patients was significantly lower compared to that of treated patients and controls: 5.33 ± 3.34 vs. 2.08 ± 0.79 ng/mL, p = 0.012, vs. 1.97 ± 1.22 ng/mL, p = 0.012, respectively. Tolvaptan-treated ADPKD patients have reduced OxSt levels compared to untreated patients. This effect may contribute to the slowing of renal function loss observed with tolvaptan treatment.