Plasma Levels Research Articles

Beneficial effects of CHF6467, a modified human nerve growth factor, in experimental neonatal hypoxic-ischaemic encephalopathy.

Therapeutic hypothermia (TH) has become the standard care to reduce morbidity and mortality in neonates affected by moderate-to-severe hypoxic-ischaemic encephalopathy (HIE). Despite the use of TH for HIE, the incidence of mortality and disabilities remains high. Nerve growth factor (NGF) is a potent neurotrophin, but clinical use is limited by its pain eliciting effects. CHF6467 is a recombinant modified form of human NGF devoid of algogenic activity (painless NGF). In rodent hippocampal slices exposed to oxygen and glucose deprivation, CHF6467 protected neurons from death and reverted neurotransmission impairment when combined with hypothermia. In a model of rat neonatal HIE, intranasal CHF6467 (20 μg kg-1) significantly reduced brain infarct volume versus vehicle when delivered 10min or 3h after the insult. CHF6467 (20 and 40 μg kg-1, i.n.), significantly decreased brain infarct volume to a similar extent to TH and when combined, showed a synergistic neuroprotective effect. CHF6467 (20 μg kg-1, i.n.) per se and in combination with hypothermia reversed locomotor coordination impairment (Rotarod test) and memory deficits (Y-maze and novel object recognition test) in the neonatal HIE rat model. Intranasal administration of CHF6467 resulted in meaningful concentrations in the brain, blunted HIE-induced mRNA elevation of brain neuroinflammatory markers and, when combined to TH, significantly counteracted the increase in plasma levels of neurofilament light chain, a peripheral marker of neuroaxonal damage. CHF6467 administered intranasally is a promising therapy, in combination with TH, for the treatment of HIE.

Using genetics to explore complement C5 as a druggable protein in periodontitis.

An excessively activated or dysregulated complement system has been proven to be a vital contributor to the pathogenesis of periodontitis. It has been previously hypothesized that inhibiting the activity of complement component C5 by targeting the C5a receptor is a powerful candidate for treating periodontitis. Here, we apply the drug target instrumental variable (IV) approach to investigate the therapeutic effect of genetically proxied inhibition of C5 on periodontitis. In our primary analysis, we used 26 independent 'cis' single nucleotide polymorphisms as IVs from the vicinity of the encoding locus of C5 that are associated with plasma C5 levels. In a secondary analysis, we assess the validity of our primary findings, exploring the involvement of alternative downstream biomarkers, interleukin 17 (IL-17), interleukin 1β (IL-1β), and tumor necrosis factor (TNF). Summary statistics of plasma levels (C5, IL-17, IL-1β, and TNF) were obtained from a genome-wide association study (GWAS) of 35,559 European descent individuals. We extracted association statistics from a GWAS of 17,353 clinical periodontitis cases and 28,210 European controls. Wald ratios were combined using inverse-variance weighted meta-analysis. In our primary approach, inhibiting C5 reduced the risk of periodontitis (Odds ratio 0.89 per 1 standard deviation reduction in C5; 95% confidence Interval 0.80-0.98, p value=0.022). Our secondary analysis suggests an involvement of IL-17 within the potential causal pathway, but was inconclusive for other biomarkers. The findings from our study suggest that C5 inhibition may reduce the risk of periodontitis, prioritizing C5 inhibitors as a potential adjunctive therapeutic intervention in this disease.

Assessment of basic pharmacokinetic parameters of dapagliflozin in TTS formulations in male minipigs

Given the extended time over which diabetes treatment is administered, the transdermal delivery system is anticipated to be a more suitable option for older individuals who may experience difficulty swallowing. The continuous delivery of dapagliflozin and more stable plasma levels are anticipated to reduce the incidence of side effects and the frequency of dosing. The objectives of the study were to determine the safety and plasma pharmacokinetics of dapagliflozin in male minipigs following application of the ointment and skin patch. In the initial phase of the study, the potential for transdermal permeation of dapagliflozin from ointment and transdermal patch to blood plasma of 15 male Göttingen minipigs was investigated. In the subsequent phase, the efficacy of utilising patches of varying strengths and sizes was assessed. The LC/MS method was employed to quantify the concentration of the active substance. The transportation of the studied API to the general circulation and accumulation in tissues were confirmed. The maximum drug concentration (122.99 ng/mL) in plasma was observed on the fourth day of application. The highest calculated Cmax was 131.91 ng/mL with a mean AUC0-last of 6620.7 ng h/mL. Following transdermal administration, dapagliflozin is excreted in the urine. The trend between urinary dapagliflozin 3-O-glucuronide levels and urinary glucose excretion was also observed. The transdermal patch has been demonstrated to be an effective drug delivery system for dapagliflozin.

SB7-H4 is a diagnostic biomarker in epithelial ovarian cancer and correlates to platinum resistance.

Ovarian cancer, with its high mortality rate among gynecological cancers, is often diagnosed late due to the lack of early diagnostic symptoms and biomarkers. The tumor immune microenvironment has become a focal point in cancer diagnostic and therapeutic research. Among these, B7-H4, a checkpoint protein, plays a crucial role in immune regulation and tumor suppression, contributing to immune evasion within the tumor microenvironment. This study aims to identify the concentration of soluble B7-H4(sB7-H4) in the plasma of patients with ovarian cancer and to evaluate its clinical significance. Through a comprehensive analysis involving enzyme-linked immunosorbent assay, immunohistochemistry, and multicolor immunofluorescence, we quantified sB7-H4 levels in patient plasma and ascites, correlating these findings with tissue expression and clinical outcomes. Results indicated a strong association between high sB7-H4 levels and advanced disease, surgical outcomes, lymphatic metastasis, and platinum resistance. When compared with traditional biomarkers CA125 and HE4, sB7-H4, especially in conjunction with these markers, enhances the diagnostic accuracy for epithelial ovarian cancer, offering insights into disease progression and therapeutic efficacy. This comprehensive analysis suggests that sB7-H4 is a promising biomarker for EOC, providing valuable insights into diagnosis, stage differentiation, treatment effectiveness, and prognosis.

SARS-CoV-2 RNAemia as a reliable predictor of long-term mortality among older adults hospitalized in pulmonary intermediate care units: a prospective cohort study

BackgroundSARS-CoV-2 viremia is associated with disease severity and high risk for in-hospital mortality. However, the impact of SARS-CoV-2 viremia on long-term outcomes in hospitalized patients with COVID-19 is poorly understood.MethodsWe conducted a prospective cohort study and recruited a group of older adult patients with COVID-19 admitted to pulmonary intermediate care units of Peking University Third Hospital during December 2022 and January 2023. The plasma level of SARS-CoV-2 RNA was determined by a standardized RT-PCR technique, and SARS-CoV-2 RNAemia was defined as a plasma viral load ≥ 50 copies/ml. In-hospital and follow-up (180-day) outcome data were collected.ResultsA total of 101 patients with an average of 80.4 years were recruited, and 63.4% of them were severe or very severe cases. Twenty-eight patients (27.7%) had SARS-CoV-2 RNAemia, with a median viral RNA load of 422.1 [261.3, 1085.6] copies/ml. Patients with SARS-CoV-2 RNAemia were more likely to develop critical cases and had a higher incidence of sepsis. Accordingly, they had a higher 180-day mortality (57.1% vs. 19.7%, P < 0.001), as well as in-hospital mortality (50.0% vs. 13.7%, P < 0.001), independent of age, disease severity, sepsis, lymphocyte count and C-Reactive protein. In addition, the risk for 180-day mortality increased with the SARS-CoV-2 RNA load in plasma. Plasma cytokines, including IL-6, IL-8 and IL-10, were higher in patients with SARS-CoV-2 RNAemia.ConclusionsOur study indicates that SARS-CoV-2 RNAemia serves as a useful biomarker for predicting mortality, especially long-term mortality, in older adult patients hospitalized in pulmonary intermediate care units.Trial registrationChinese Clinical Trial Registry website (No. ChiCTR2300067434).

Plasma glucose, HbA1c, insulin and lipid profile in Sudanese type 2 diabetic patients with cardiovascular disease: a case control study

Background: Type 2 diabetes mellitus (T2DM) and its consequences are a serious global public health issue. By 2030, the number of people with type 2 diabetes is predicted to reach 439 million. The purpose of this study is to evaluate the plasma levels of glucose, HbA1c, insulin, and lipid profile in Sudanese T2DM patients. Methods: This case control study included 165 Sudanese patients with diabetic type 2 and a cardiovascular condition as cases and 165 diabetic type 2 volunteers without a cardiovascular disorder as controls. The concentrations of plasma glucose, HbA1c, and lipid profile were assessed using a Mindray BS-480 auto-chemistry analyzer, and insulin was analyzed using a Cobase 411 auto analyzer. The collected data were analyzed using statistical tools for social science computer programs (SPSS version 21). Results: According to the findings, (59.4%) of patients between the ages of (50-69). Females made up 50.9%. (38.2%) of patients had an illness duration of between (8-15 years). (41.8%) of individuals did not have hypertension. There was a substantial rise in BMI, FBG, HbA1c, HDL-C, and insulin among diabetics with cardiovascular disease compared to diabetics without cardiovascular disease (p-value = 0.001, 0.000, 0.018, and 0.000). Females had significantly higher blood TC, LDL-C, HDL-C, and BMI than males (p-values = 0.000, 0,001, and 0.000, respectively). There were significant positive correlation between FBS, HBA1c, insulin and duration of disease (r=0.155, p, value=0.005) (r=0.160, p, value=0.004)(r=0.103, p. value=0.061)respectively, while there were significant negative correlation between TC, TG,LDL-C, HDL-C and duration of disease (r=-0.152, p, value= 0.006)(r=-0.023, p, value=0.678)(r=-0.113, p, value= 0.040)(r=-0.145, p, value=0.008)respectively. Conclusion: When comparing diabetics with cardiovascular disease to diabetics without cardiovascular disease, there was a substantial rise in BMI, FBG, HbA1c, HDL-C, and insulin. FBS, HBA1c, insulin, and illness duration all had a strong positive connection.

Vitamin D deficiency may increase the risk of acute kidney injury in patients with diabetes and predict a poorer outcome in patients with acute kidney injury

BackgoundPeople with diabetes are much more likely to develop acute kidney injury (AKI) than people without diabetes. Low 25-hydroxy-vitamin D [25(OH)D] concentrations increased the risk of AKI in specific populations. Few studies have explored the relationship between the 25(OH)D level and AKI in patients with diabetes. We conducted this study to investigate the relationship between the plasma level of 25(OH)D and the risk of AKI in patients with diabetes, and to evaluate whether the 25(OH)D level could be a good prognostic marker for AKI progression.MethodsA total of 347 patients with diabetes were retrospectively reviewed. The primary endpoint was the first event of AKI. The secondary endpoint is need-of-dialysis. AKI patients were further followed up for 6 months with the composite endpoint of end-stage renal disease (ESRD) or all-cause death. Kaplan–Meier survival analysis and Cox proportional hazards models were used.ResultsDuring a median follow-up of 12 weeks (12.3 ± 6.7), 105 incident AKI were identified. The middle and high tertiles of baseline 25(OH)D levels were associated with a significantly decreased risk of AKI and dialysis compared to the low tertile group (HR = 0.25, 95% CI 0.14–0.46; HR = 0.24, 95% CI 0.13–0.44, respectively, for AKI; HR = 0.15; 95% CI 0.05–0.46; HR = 0.12; 95% CI 0.03–0.42, respectively, for dialysis). Sensitivity analysis revealed similar trends after excluding participants without history of CKD. Furthermore, AKI patients with 25(OH)D deficiency were associated with a higher risk for ESRD or all-cause death (HR, 4.24; 95% CI, 1.80 to 9.97, P < 0.001).ConclusionA low 25 (OH) vitamin D is associated with a higher risk of AKI and dialysis in patients with diabetes. AKI patients with 25(OH)D deficiency were associated with a higher risk for ESRD or all-cause death.

Improving nutrient digestibility and health in rabbits: effect of fermented rapeseed meal supplementation on haematological and lipid parameters of blood

BackgroundMany studies conducted on livestock point to fermented rapeseed meal (FRSM) as a component that provides adequate quality and quantity of protein. Additionally, it is a very good source of probiotics, prebiotics, enzymes, and antioxidants. A study was undertaken to assess the impact of a feed supplemented with FRSM fermented with Bacillus subtilis strain 87Y on production parameters, nutrient digestibility, and haematological and lipid indicators of the blood in growing rabbits. Forty New Zealand White rabbits (body weight 816,25 ± 24,98 g) aged 35 days were used in this study. The animals were divided into four groups, a control group (C) was fed a standard diet, while in the diet of three experimental groups, soybean meal (SBM) was replaced with FRSM at 4% (FR4 group), 8% (FR8 group), or 12% (FR12 group).ResultsDetailed analysis revealed that the contribution of FRSM in the rabbit feed, regardless of the amount, had a significant effect on body weight gain (BWG), as well as average daily gain (ADG), compared to the control group (C), (P = 0.017). All groups receiving FRSM had a significantly lower (P = 0.05) feed conversion ratio (FCR) compared to the control group. In rabbits fed a diet containing 8% FRSM, the blood haematological parameters, such as red blood cells (RBC), haematocrit (HCT), haemoglobin (HGB), and mean corpuscular volume (MCV), were significantly higher compared to the control group (P = 0.037). In addition, a significant reduction in the plasma levels of the low-density lipoprotein fraction (LDL-chol), the ratio of total cholesterol to high-density lipoprotein (CHOL/HDL), (P = 0.001), and triacylglycerols (TG), (P = 0.004) were observed in the experimental groups compared to the control group.ConclusionsBased on the encouraging outcomes, it is possible to recommend domestically produced FRSM as a viable substitute for genetically-modified (GM) SBM in rabbit feed.

Plasma and CSF neurofilament light chain distinguish neurodegenerative from primary psychiatric conditions in a clinical setting.

People with neurodegenerative disorders (ND) frequently face diagnostic delay and misdiagnosis. We investigated blood and cerebrospinal fluid (CSF) neurofilament light chain (NfL) to distinguish ND from primary psychiatric disorders (PPD), a common challenge in clinical settings. Plasma and CSF NfL levels were measured and compared between groups, adjusting for age, sex, and weight. A total of 337 participants were included: 136 ND, 77 PPD, and 124 Controls. Plasma NfL was 2.5-fold elevated in ND compared to PPD and had strong diagnostic performance (area under the curve, [AUC]: 0.86, 81%/85% specificity/sensitivity) that was comparable to CSF NfL (2-fold elevated, AUC: 0.89, 95%/71% specificity/sensitivity). Diagnostic performance was especially strong in younger people (40-<60 years). Additional findings were cutoffs optimized for sensitivity and specificity, and issues important for future clinical translation. This study adds important evidence for a simple blood-based biomarker to assist as a screening test for neurodegeneration and distinction from PPD, in clinical settings. NfL levels were significantly higher in ND versus PPD. Plasma NfL showed strong diagnostic performance, comparable to CSF NfL, to distinguish ND from PPD. Diagnostic performance was higher in younger people, where diagnostic challenges are greater. Further research is needed on analytical and reference range factors, for clinical translation. These findings support a simple screening blood test for neurodegeneration.

Role of Plasma Angiopoietin-1 and VEGF Levels as Potential Biomarkers in Chronic Central Serous Chorioretinopathy with Macular Neovascularization.

To evaluate the plasma levels of angiopoietin-1 and vascular endothelial growth factor (VEGF) and their association with macular neovascularization (MNV) in patients with chronic central serous chorioretinopathy (cCSC). Correlations between plasma cytokine levels, CSC duration, and mean choroidal thickness (CT) were also investigated. Of the 59 patients with cCSC, 10 patients with MNV secondary to cCSC and 10 patients with cCSC without MNV were enrolled in the study. The control group included 15 healthy volunteers matched for age, sex, smoking status, and comorbidities. Chronic CSC was diagnosed based on typical findings on swept-source optical coherence tomography (OCT), fundus fluorescein angiography, and indocyanine green angiography. Additionally, all patients underwent OCT angiography to help detect MNV. Plasma angiopoietin-1 and VEGF levels were assessed using multiplex immunoassay. The plasma angiopoietin-1 levels differed between the 3 groups (p = 0.005). The angiopoietin-1 levels were lower in patients with cCSC with MNV than in controls (p = 0.006). There were no differences in the plasma VEGF levels between all the 3 groups (p = 0.329). The VEGF levels were negatively correlated with mean CT in cCSC patients with MNV (rho = -0.683, p = 0.042) but correlated positively with disease duration in patients with cCSC without MNV (rho = 0.886, p = 0.003). Our study confirms that MNV is a common complication of cCSC and provides new insights into the role of angiopoietin-1 in cCSC and MNV. Reduced angiopoietin-1 levels in cCSC patients, regardless of MNV status, highlight the importance of the Ang-Tie2 pathway in disease pathogenesis and may point to new therapeutic targets and future novel treatments to improve the management of these patients.

Current summer heat waves impair rainbow trout (Oncorhynchus mykiss) spermatogenesis: Implications for future fish farming management practices in South Europe

Freshwater environments provide different ecosystem services for human well-being, and are one of the most important natural sources of healthy animal protein. Climate change and extreme temperature events, among other factors, represent a growing concern for their sustainable use. In contrast to the research effort to elucidate how global warming affects these aquatic environments, few studies pay attention to the potential effects of heat waves, a current phenomenon with direct effects on contemporaneous organisms. Here, the potential effects of a single heat wave event on rainbow trout (Oncorhynchus mykiss) males, mimicking a recorded environmental event, have been evaluated by means of biometric, redox and stress response, plasma testosterone, histopathology and transcriptomic analyses. Environmental monitoring showed how, during the last 4 years, mean water temperature increased (2–4 °C; from April to October), heat wave duration progressively increased (from 15 to 100 days), and mean intensities ranged 1.7–3.8 °C with a maximum of 8.0 °C. An experimentally recreated heat wave lasting only 16 days, with mean and maximum intensities of 2.7 and 6.7 °C, induced a decrease in body weight and total antioxidant status, but increased blood plasma cortisol levels and redox enzyme activity. The induced heat wave decreased testosterone plasma levels and sperm quality, arrested spermatocyte type I and II differentiation and increased spermatozoa apoptosis. These effects were correlated with a differential expression of 1156 genes (116 up- and 1040 down-regulated genes) in exposed fish when compared to Control fish, reflecting an alteration in cell apoptosis and spermatozoa maturation processes. These results not only increase concern about current heat wave events, recreating a real scenario, but also unveil for the first time the particular molecular mechanisms of subsequent spermatogenesis disruption, opening new avenues for designing urgent and necessary strategies to mitigate the effects of extreme climate change conditions on freshwater aquaculture.

Plasma levels of polyunsaturated fatty acids and multiple sclerosis susceptibility in a US case-control study

BackgroundThere are plausible mechanisms, yet mixed evidence, that higher polyunsaturated fatty acids (PUFAs) levels reduces the risk of multiple sclerosis (MS). Prior studies relied on dietary surveys to estimate levels. ObjectiveWe tested associations between plasma levels of n-3 and n-6 PUFAs and likelihood of MS onset or clinically isolated syndrome (CIS) using data from the MS Sunshine Study, a case-control study conducted in the United States. MethodsCase participants (n = 589) aged ≥ 18 years and matched control participants (n = 630) were recruited between 2011 and 2015. Plasma phospholipid fatty acid profiling was conducted by gas-liquid chromatography. We used logistic regression to report odds ratios, testing for interactions, adjusting for covariates and correcting for multiple comparisons. ResultsThere was a 6 % lower probability of MS/CIS per unit increase in total n-6 PUFA level, expressed as a percentage of total plasma phospholipid fatty acids (odds ratio = 0.94; 95 % confidence interval = 0.90,0.98; p = 0.012). We found no statistically significant association between individual or total plasma levels of n-3 PUFAs and probability of MS/CIS; however, plasma levels of n-3 PUFAs were low across the cohort. No other individual or aggregate PUFA levels were significantly associated with MS/CIS. ConclusionA higher total n-6 PUFA level may be beneficial in terms of MS susceptibility.Further research is needed to determine whether n-3 PUFAs may be beneficial only above a threshold that is achievable by supplementation.

A New Plant Active Polysaccharide from Nicotiana Improves the Lead-Led Impairment of Spatial Memory in Mice by Modulating the Gut Microbiota and IL-6.

Active polysaccharides from plants are broadly applied in the food and health industry. The purpose of this study is to identify a new plant active polysaccharide and to investigate its role in modulating spatial memory. Ultrasonics and DEAE-52 chromatography were used to separate and purify the plant active polysaccharide (PAP). Mice were exposed to 100 ppm of lead acetate from birth to 7 weeks old to establish the memory impairment model. PAPs with concentrations of 200 or 400 ppm were fed to the subject mice each day after weaning in a spatiotemporally separated fashion. At the end of the intervention, mice were examined using the Morris water maze test, microbiome sequencing, cytokine profiling and protein analysis. The derived active polysaccharide was constituted by β-anomeric carbon, indicating a new form of PAP. The PAP significantly ameliorates the memory impairment caused by postnatal lead exposure, as evidenced by the preferred coverage of the test mouse in the hidden platform, demonstrating salient neuroregulatory activity. In terms of the gut microbiome in response to PAP treatment, it was found that the 400 ppm PAP reversed the gut dysbiosis, producing a comparable structure to the intact animals, represented by the relative abundance of Firmicutes and Muribaculum, Desulfovibrio, etc. For cytokines, the PAP reversed the plasma levels of IL-6, suggesting an anti-inflammatory trend in the context of proinflammation caused by lead invasion. By injecting an IL-6 antagonist, Tocilizumab, into the deficient mice, the spatial memory was significantly repaired, which demonstrates the central roles of IL-6 in mediating the positive effect of the PAP. Finally, a histone modification mark, H3K27me3, was found to be potent in responding to the signals conveyed by the PAP. The PAP could improve the memory deficits by remodeling the gut-brain axis centered at the microbiota and IL-6, which is regarded as an important cytokine-modulating brain activity. This is an intriguing instance linking neuromodulation with the active polysaccharide, shedding light on the innovative applications of plant polysaccharides due to the scarcity of similar phenotypic connections.

8536 Unusual Finding of Familial Adenomatous Polyposis in a Patient with X Linked Adrenoleukodystrophy

Abstract Disclosure: A. Deswal: None. A. Dwarakanathan: None. Introduction: X Linked Adrenal Leukodystrophy (X-ALD) is a rare hereditary genetic disorder of impaired peroxisomal metabolism due to a defective transmembrane protein, ADLP. Defective ADLP impairs transport of VLCFA-CoA esters across cytosol into peroxisomes leading to build-up of toxic VLCFA in the cells. X-ALD is caused by mutation in ABCD1 gene, located on X chromosome, which codes for ADLP. It has a broad clinical spectrum including Cerebral ALD, Adrenomyeloneuropathy and Adrenocortical insufficiency. X-ALD has a general prevalence of 1 in 17,000 newborns. Approximately 80% of males with X-ALD develop primary adrenal insufficiency in their lifetime. It can be diagnosed by measuring VLCFA levels in plasma , checking for ABCD1 gene mutation or by doing Brain MRI (depending upon phenotypical presentation ). Case presentation: We report a case of a 23 year old man who had primary adrenocortical insufficiency subtype of X-ALD. His mother was a carrier of ABCD1 gene mutation. He himself tested hemizygous for ABCD1 gene. He came to our office for a follow up visit after an episode of adrenal crisis for which he was hospitalized. He was started on replacement therapy with prednisone and fludrocortisone. Over the next couple of years, he was diagnosed with diabetes mellitus with initial HbA1C of 8.8%. GAD antibody test for type I DM came out negative. His diabetes was managed with metformin and pioglitazone( later discontinued) .By the age of 25,he was found to have polyposis of colon. He tested positive for APC gene mutation . He underwent a J pouch surgery . He also developed PICA aneurysm. Discussion: X-ALD is a hereditary progressive disease with no association between genetic and phenotypic presentations. Adrenal insufficiency type is usually the first to present. Cerebral ALD is typically diagnosed between the ages of 3-18 presenting as psychiatric illness. Myelopathy typically starts by middle age and involves the spinal cord, causing the affected individuals to develop gait disturbances , bladder and bowel dysfunction, with severe mobility dysfunction by 5th-6th decade of life. We present a unique case of a young man with adrenal insufficiency type of X-ALD who also developed gastrointestinal pathology. Per our literature review, X-ALD has not been seen associated with APC gene mutations or colon carcinomas . Pathology behind XLD revolves around ABCD1 gene mutaion causing VLCFA accumulation in plasma, fibroblasts, adrenal glands, oligodendrocytes , astrocytes etc. Whereas FAP involves APC gene mutation which affects the intestinal epithelial cells leading to their apoptosis and polyp formation. X-ALD and FAP are disorders of two different gene mutations with varied clinical presentations. But the question arises regarding finding of APC gene mutation in a patient with ABCD1 gene mutation that if it is a mere co-incidence or common mutation pathway linking these two disorders exists. Presentation: 6/2/2024

8500 Fvb/Ant Mice Carrying the Thr92Ala-Dio2 Polymorphism Have a Goiter

Abstract Disclosure: A. Batistuzzo: None. B. Bocco: None. E. McAninch: None. M.O. Ribeiro: None. A.C. Bianco: Consulting Fee; Self; Abbvie. Fvb/Ant Mice Carrying the Thr92Ala-Dio2 Polymorphism Have a Goiter Introduction: The Thr92Ala-DIO2 polymorphism is prevalent worldwide, with about 50% of the population carrying at least one polymorphic allele. Ala-D2 is ectopically located in the Golgi apparatus and causes endoplasmic reticulum (ER) stress. It is also about 40 % less active than Thr92-D2, thus T3 production could be compromised in carriers of the polymorphism. Objective: C57BL/6J (B6) mice carrying the Ala92-DIO2 polymorphism exhibit no significant thyroid phenotype as compared to control Thr92-Dio2 mice. Here we wished to test whether changing the genetic background of mice can influence the thyroidal impact of the Thr92Ala-DIO2 polymorphism. Therefore, we backcrossed B6-Thr92Ala-Dio2 mice to the FVB/Ant background for 8 generations. Methods: Mice were killed by asphyxiation in a CO2 chamber and blood collected from 7-18 weeks old female and male FVB/Ant Ala-Dio2 mice to measure plasma levels of TSH, T4, T3, and rT3. The thyroid gland was dissected, weighed and processed for histological analysis. The weight of the thyroid gland was normalized by femoral length. The QuPath Software was used for measuring follicular area, cell height, and the number of internalized follicular cells. Thr92-Dio2 mice of the same age and sex were used as controls. Data were analyzed using PRISM software and expressed as mean±SE (TH plasma levels and thyroid weight) or mean±SD (thyroid histology). A Student t-test was used to compare 2 groups. Results: Male and female Ala mice had significantly higher levels of TSH (males: 0.36±0.04 vs 1.1±0.21 ng/ml; p&amp;lt;0.01 and females: 0.017±0.006 vs 0.12±0.06 ng/ml; p&amp;lt;0.01), lower levels of plasmatic T4 (males: 2.9±0.2 vs 1.3±0.25 µg/dl; p&amp;lt;0.01 and females: 2.1±0.16 vs 1.15±0.16 µg/dl; p&amp;lt;0.001) and lower levels of rT3 (males: 23.8±2 vs 14.2±1.3 ng/dl; p&amp;lt;0.001 and females: 16.45±1.4 vs 10.4±1.17 ng/dl; p&amp;lt;0.01). Additionally, the thyroid gland of female polymorphic mice was heavier (4.3±0.39 vs 1.8±0.17 mg/cm; p&amp;lt;0.001) and both sexes presented enlarged thyroid follicle area (males: 1464±1464 vs 2055±2443 µm2; p&amp;lt;0.0001 and females: 1519±1364 vs 2597±2624 µm2; p&amp;lt;0.0001) and a higher number of internalized follicular cells (females: 0.06±0.007 vs 0.17±0.008 internalized cells/follicle; p&amp;lt;0.0001). No differences in the follicular cell height were observed. Discussion: In contrast to B6, FVB mice carrying the Thr92Ala-D2 polymorphism exhibit goiter, increased follicular area, slightly reduced serum T4 levels, and elevated serum TSH levels. These findings suggest that the impact of the Thr92Ala polymorphism varies according to the genetic background. They may explain why clinical studies of patients carrying the Thr92Ala-DIO2 polymorphism have yielded inconsistent results when different populations are analyzed. Presentation: 6/3/2024

CD151 identifies an NK cell subset that is enriched in COVID-19 patients and correlates with disease severity

Severe coronavirus disease 2019 (COVID-19) often leads to acute respiratory distress syndrome and multi-organ dysfunction, driven by a dysregulated immune response, including a cytokine storm with elevated proinflammatory cytokine levels. Natural killer (NK) cells are part of the innate immune system with a fundamental role in the defense against viral infections. However, during COVID-19 acute infection, they exhibit an altered phenotype and impaired functionality contributing to the immunopathogenesis of the disease. In this work, we have studied a cohort of patients with COVID-19 (ranging from mild to severe) by analyzing IL-15, TGF-β, PlGF and GDF-15 plasma levels and performing multiparametric flow cytometry studies. Our results revealed that severe COVID-19 patients exhibited high levels of IL-15, PlGF and GDF-15, along with an enrichment of anNK cell subset expressing the CD151 tetraspanin, which correlated with IL-15 plasma levels and disease severity. In patients, these CD151+ NK cells displayed a more activated phenotype characterized by an increased expression of HLA-DR, CD38 and granzyme B, a distinct receptor repertoire, with lower levels of CD160 and CD31 and higher levels of CD55 and, remarkably, a higher expression of tissue-resident markers CD103 and the NK cell decidual marker CD9. Last of all, in individuals with severe disease, we identified an expansion of a CD151brightCD9+ NK cell subset, suggesting that these cells play a specific role in COVID-19. Altogether, our findings suggest that CD151+ NK cells may have a relevant role in COVID-19 immunopathogenesis.

8691 Recovery Of Adrenal Function After Stopping Mitotane In Patients With Adrenocortical Carcinoma

Abstract Disclosure: B. Altieri: None. O. Kimpel: None. F. Megerle: None. M. Detomas: None. I.O. Chifu: None. C.T. Fuss: None. M. Quinkler: None. M. Kroiss: None. M. Fassnacht: None. Background: Mitotane is the standard therapy of adrenocortical carcinoma (ACC), both adjuvantly in patients with high risk of recurrence and palliative setting, due to its relative selectivity of its cytotoxic effects towards adrenocortical cells. Therefore, it virtually always leads to adrenal insufficiency. However, frequency and characteristics of hypothalamic-pituitary-adrenal (HPA) axis recovery after discontinuation are not well defined. Methods: Retrospective study of patients with ACC adjuvantly treated with mitotane for ≥12 months who were disease-free at mitotane stop and had a minimum follow-up ≥1 year. Data on patients and tumor characteristics, mitotane treatment, and information on HPA axis were analyzed. Primary endpoint was the adrenal recovery. Explorative analysis of predictive factors (e.g. sex, age, follow-up in reference center, cumulative mitotane dose and plasma levels, duration of treatment, and dose of hydrocortisone-equivalent replacement) was performed using Cox regression. Mitotane plasma elimination rate and hormonal changes after mitotane stop were also investigated. Results: 56 patients (36 women) treated with mitotane for a median time of 25 months and an average daily dose of 2.8 g (interquartile range 1.8-3.4) were included. The average hydrocortisone-equivalent replacement daily dose during mitotane treatment was in median 49.4 mg (41.1-53.5). Mitotane plasma levels decreased slowly after discontinuation, but with a very high variability between individual patients. Median time until mitotane levels dropped below 5 mg/L, 2 mg/L, and the detection limit was 152 days (114-202), 280 days (192- 37 370), and 395 days (227-546), respectively. Full adrenal recovery was documented in 32 (57%) patients after a median time of 26 months (95%CI=19.6-32.4). Among these, 22 (69%) achieved HPA recovery within 24 months. To note, a complete recovery after more than 67 months did not occur. Partial and insufficient recovery were observed in 10 (18 %) and 14 (25%) patients, respectively. In four patients (7.1%) adrenal insufficiency persisted &amp;gt;5 years after discontinuation. Mitotane peak ≥27mg/L significantly correlated with longer time to adrenal recovery (HR=0.2, 95%CI=0.1-0.8, p=0.03). 27/38 patients (71%) followed in reference centers achieved adrenal recovery compared to only 5/18 (28%) followed-up in non-reference centers (HR=4.51, 95%CI=1.71-11.89, p=0.002). Other investigated factors were not associated with adrenal function after discontinuation. Conclusions: Our study demonstrates that adrenal recovery occurs in most patients after stopping mitotane, particularly when followed-up in specialized centers, but not in all. Elimination time of mitotane after treatment discontinuation is very long, but individually quite variable. Presentation: 6/2/2024

8577 Gonadal function throughout life in XX and XY patients with a germline WT1 variant - lessons from a cohort of 80 patients

Abstract Disclosure: M. Carré Lecoindre: None. D. Mallet: None. C. Dossier: None. M. Glenisson: None. M. Grapin: None. A. Brac de la Perriere: None. Z. Chakhtoura: None. C. Bouvattier: None. M. Houang: None. C. Pienkowski: None. N. Zaegel: None. R. Rayneau,MD and PhD: None. T. Blanc: None. L. Martinerie: None. The WT1 gene is involved in tissue homeostasis, tumorigenesis, as well as embryonic development of kidneys, bipotential gonads and ovarian determinism. Germline WT1 variants are known to generate kidney and gonadal developmental diseases associated with differences in genital development (DSD). However, the state of gonadal function, its evolution over time, and the impact of WT1 variants on pubertal development and fertility potential in patients have never been studied. A French national, retrospective, multi-center, observational study evaluated gonadal function over time in patients with a germline variant of the WT1 gene, according to XX or XY karyotype and genotype. Among the 80 patients included - the largest cohort to date - 30% had a XX karyotype, 95% of whom were assigned female at birth. Importantly, 40% of them had uterine malformation, which was not known to be associated with WT1 anomalies. Patients with a XY karyotype presented with DSD (59%), typical female genitalia (15%) or typical male genitalia with or without bilateral cryptorchidism (27%). Thirty percent of XX and 87% of XY patients had gonadal dysgenesis defined as gonads with a dysplastic and/or dysmorphic appearance on histology, surgery and/or imaging or biological hormonal markers compatible with gonadal insufficiency. However only 3 patients (4%) developed gonadal tumors to date. Spontaneous puberty was observed in 94% of XX and 60% of XY patients. However, 67% had impaired gonadal function - defined as AMH and/or inhibin B below the threshold for age and/or elevated FSH and/or LH above 10 UI/L - at the age of 12.6 years and 83% at the age of 16.2 years, considering all karyotypes and genotypes. Hormonal treatment had to be initiated in 57% of XY patients with a median age at 14.4 years old. No difference was seen between patients with or without renal failure (p-value = 0,37), nor regarding the timing of renal insufficiency. In addition, increased FSH, LH and decreased AMH or inhibin B plasma levels were observed independently from gonadotoxic treatment exposure in most patients. None of the patient has had children to date.In conclusion, most XX and XY patients with a germline variant of WT1 gene are affected by early gonadal insufficiency of variable severity with potential impact on puberty and fertility. Even though dialysis, immunosuppressors, gonadotoxic chemotherapies, local radiotherapy and gonadal surgery are known to impair gonadal function, the gonadal disease linked to WT1 variants is the principal factor of gonadal impairment. For these reasons, regular follow-up by an endocrinologist appears essential for both XY and XX patients, providing clear information about gonadal function and fertility. Presentation: 6/1/2024

7881 Using Selective Estrogen Receptor Modulators to Reveal Therapeutic Vulnerabilities in ESR1 Mutant Breast Cancer

Abstract Disclosure: S.W. Fanning: Grant Recipient; Self; Olema Oncology. Hotspot activating somatic mutations to ESR1, the gene for estrogen receptor alpha (ER), enable hormone therapy resistance and drive breast cancer metastasis. The Y537S missense mutation within the ER ligand binding domain is the most activating, therapy resistant, and increases metastatic potential. Next generation antiestrogens including lasofoxifene (laso) and elacestrant (RAD1901) show improved progression-free survival over fulvestrant (fulv), but further progress is needed to achieve durable response. To fully address Y537S ESR1, it is critically important to understand how the mutation boosts metastasis. We have developed a small molecule antiestrogen, T6I-29, that uniquely impacts transcriptional pathways related to cell-cell adhesions and morphology. Two allele-specific phenotypes identified by the Oesterreich Laboratory enhance the metastasis of Y537S ESR1 breast cancer cells. T6I-29-specific pathway effects were driven by a significant downregulation of dickkopf-1 (DKK1), which was also observed but to a lesser extent with fulv, laso, and RAD1901. DKK1 is an immunosuppressive, tumor-secreted glycoprotein and its overexpression correlates with poor outcomes in multiple cancers. However, DKK1 acts as a tumor-suppressor in colorectal cancer, which does not rely on ER. Our preliminary studies suggest that DKK1 plays a role in ER+ breast cancer progression. We measured significantly elevated DKK1 protein levels in the blood plasma of 108 diverse ER+ breast cancer patients compared to 100 matched healthy female donors and levels increase coincident with tumor stage. Unfortunately, we lack ESR1 mutation status and other critical details to make further insights on these patients. DKK1 gene expression and protein secretion is stimulated in WT cells by the pathogenic hormone 17β-estradiol (E2), while it is constitutively expressed in Y537S ESR1 cells. DKK1 is characterized as a WNT antagonist but can also induce PI3K/Akt signaling. Interestingly, DKK1 is elevated and ERα responsive in WT not mutant PI3KCA cell lines. DKK1 knockdown also reduced proliferation of Y537S ESR1 breast cancer cells where elevated DKK1 was observed. Presentation: 6/3/2024

6824 Bile Acid Elevation in Nonalcoholic Steatohepatitis Correlates With Fibrogenesis and Mitochondrial Dysfunction In Vitro

Abstract Disclosure: N. Patel: None. Y. Li: None. M. Raul: None. C. Sottas: None. V. Papadopoulos: None. Nonalcoholic steatohepatitis (NASH) is the most severe form of nonalcoholic fatty liver disease. It is estimated that over 115 million adults are affected by NASH worldwide. The causes of NASH are unclear, and no treatment is yet available. Bile acids (BAs) are steroids synthesized in the liver, promoting lipid absorption. The levels of BAs are tightly regulated in the body. Patients diagnosed with NASH frequently exhibit increased levels of BAs in both liver tissue and plasma, indicating a potential association between elevated BA concentrations and the pathogenesis of NASH. In an in vivo study using a high-fat diet rat model of NASH, we measured plasma BA composition and levels. The results obtained indicated substantial increases in the concentrations of glycocholic acid (GCA), glycochenodeoxycholic acid (GCDCA), glycodeoxycholic acid (GDCA), and taurodeoxycholic acid (TDCA) in NASH compared to controls. These findings suggested their potential implication in the development of NASH. To elucidate the mechanistic link between changes in BA synthesis and fibrogenesis, in vitro experiments were conducted using the LX-2 humane hepatic stellate and Huh7 human hepatocellular carcinoma cells. First, the concentrations of the selected bile acids (GCA, GCDCA, GDCA, and TDCA) used in our studies were found to be non-toxic to the cells using the MTT assay. Subsequent qPCR analyses demonstrated the upregulation of key fibrotic genes (TGFβ, COL1A1, ACTA2) in LX-2 cells following exposure to individual bile acids GCA, GCDCA, GDCA, and TDCA. Immunocytochemistry (ICC) studies further demonstrated fibrogenesis by revealing increased expression of COL1A1 following treatment with each of the four bile acids. These data suggest that changes in BA formation are linked to fibrogenesis. We subsequently explored the impact of BAs on mitochondrial function using the Seahorse XF Cell Mito Stress Test in Huh7 cells. The results obtained revealed a reduction in the oxygen consumption rate and ATP production in response to BA treatment. This observation suggests a potential role for modified BAs in disrupting mitochondrial homeostasis, a recognized contributor to the progression of NASH. In summary, the data presented provide valuable insights into the complex effects of modified BAs on NASH pathogenesis, establishing connections between changes in BA composition and fibrogenesis, mitochondrial dysfunction. Presentation: 6/2/2024