Differential Peripheral Blood Research Articles

Tissue-specific sex differences in pediatric and adult immune cell composition and function.

Sex-based differences in immune cell composition and function can contribute to distinct adaptive immune responses. Prior work has quantified these differences in peripheral blood, but little is known about sex differences within human lymphoid tissues. Here, we characterized the composition and phenotypes of adaptive immune cells from male and female ex vivo tonsils and evaluated their responses to influenza antigens using an immune organoid approach. In a pediatric cohort, female tonsils had more memory B cells compared to male tonsils direct ex vivo and after stimulation with live-attenuated but not inactivated vaccine, produced higher influenza-specific antibody responses. Sex biases were also observed in adult tonsils but were different from those measured in children. Analysis of peripheral blood immune cells from in vivo vaccinated adults also showed higher frequencies of tissue homing CD4 T cells in female participants. Together, our data demonstrate that distinct memory B and T cell profiles are present in male vs. female lymphoid tissues and peripheral blood respectively and suggest that these differences may in part explain sex biases in response to vaccines and viruses.

Peripheral T lymphocyte and immunocyte subset dynamics: markers of neoadjuvant therapy outcomes in esophageal squamous cell carcinoma.

In patients with resectable esophageal squamous cell carcinoma (ESCC), neoadjuvant therapy increased the curative resection rate, disease-free survival, and overall survival for patients with resectable ESCC. However, the efficacy of neoadjuvant therapy varies among different patients. We aim to compare the differences in the characteristics of peripheral blood T lymphocyte subsets before and after neoadjuvant therapy in patients with different curative efficacy. This study enrolled 266 ESCC patients who received neoadjuvant therapy and esophagectomy from August 2018 to August 2022. The postoperative pathological results divided patients into the major pathological response (MPR) and non-MPR groups. Compare the differences in peripheral blood T lymphocyte subsets and analyze the trend of changes in T lymphocyte subsets at different phases of treatment. Propensity score matching was used to reduce the influence of potential confounding factors. Prior to the neoadjuvant therapy, particularly before the second cycle, the MPR group exhibited significantly higher ratios of CD4/CD8 (P=0.009) and helper T cells (TH ratio, P=0.030) compared to the non-MPR group. In contrast, the suppressor T cell ratio (TS ratio) was lower (P=0.016) in the MPR group. The difference in peripheral blood lymphocyte subsets between the two groups of patients who underwent neoadjuvant chemoradiotherapy is significant. In peripheral blood, T lymphocyte subsets varied significantly based on the effectiveness of neoadjuvant treatment. Prior to the second cycle of neoadjuvant therapy, a higher CD4/CD8 and TH ratio, coupled with a decreased TS ratio, might suggest enhanced treatment outcomes.

343. T LYMPHOCYTE SUBSETS CHARACTERISTICS IN DIFFERENT CURATIVE EFFICACY OF NEOADJUVANT THERAPY FOR PATIENTS WITH ESOPHAGEAL SQUAMOUS CELL CARCINOMA

Abstract Background In patients with resectable esophageal squamous cell carcinoma(ESCC), neoadjuvant therapy increased the curative resection rate, disease-free survival, and overall survival. Up to 32% of patients with esophageal cancer show a pathological complete response after neoadjuvant therapy. However, the efficacy of neoadjuvant therapy varies among different patients. We aim to compare the differences in the characteristics of peripheral blood T lymphocyte subsets before and after neoadjuvant therapy in patients with different curative efficacy. Methods This study enrolled 269 ESCC patients who received neoadjuvant treatment followed by esophagectomy planned 4–6 weeks after preoperative therapy. The postoperative pathological results divided patients into the main pathological response (MPR) and non-MPR groups. Compare the differences in peripheral blood T lymphocyte subsets before neoadjuvant therapy, and analyze the trend of changes in T lymphocyte subsets before neoadjuvant therapy, before surgery, in the short term after surgery, and at the initial follow-up after surgery. Results Before receiving neoadjuvant therapy, the peripheral blood CD4/CD8(p = 0.001) and helper T cell rate (THrate)(p = 0.006) in the MPR group were significantly higher than those in the non-MPR group. In contrast, the suppressor T cell rate(TSrate) was lower(p = 0.029) in the MPR group. After neoadjuvant therapy, the CD4/CD8 in both groups reached similar classes, but the THrate decreased dramatically and TSrate increased more in the MPR group. At the first follow-up after surgery, the peripheral blood CD4/CD8, THrate, and TSrate of both groups of patients tended to be at the same level. Conclusion T lymphocyte subsets in the peripheral blood significantly differed among patients with different neoadjuvant treatment efficacy. High levels of CD4/CD8, THrate, and low levels of TSrate before receiving neoadjuvant therapy may have better curative efficacy.

Non-targeted Metabolomics Profiling of Plasma Samples From Patients With Major Depressive Disorder.

BackgroundMajor depressive disorder (MDD) is a neuropsychiatric disorder caused by multiple factors. Although there are clear guidelines for the diagnosis of MDD, the direct and objective diagnostic methods remain inadequate thus far.MethodsThis study aims to discover peripheral biomarkers in patients with MDD and promote the diagnosis of MDD. Plasma samples of healthy controls (HCs, n = 52) and patients with MDD (n = 38) were collected, and then, metabolism analysis was performed using ultrahigh-performance liquid chromatography–tandem mass spectrometry (LC–MS/MS). Heatmap analysis was performed to identify the different metabolites. Meanwhile, receiver operating characteristic (ROC) curves of these differential metabolites were generated.ResultsSix differential metabolites were found by LC–MS/MS analysis. Three of these were increased, including L-aspartic acid (Asp), diethanolamine, and alanine. Three were decreased, including O-acetyl-L-carnitine (LAC), cystine, and fumarate. In addition, LAC, Asp, fumarate, and alanine showed large areas under the curve (AUCs) by ROC analysis.ConclusionThe study explored differences in peripheral blood between depressed patients and HCs. These results indicated that differential metabolites with large AUCs may have the potential to be promising biomarkers for the diagnosis of MDD.

Comparison of the effects between MPL and JAK2V617F on thrombosis and peripheral blood cell counts in patients with essential thrombocythemia: a meta-analysis

To assess the effects between MPL and JAK2V617F on the thrombosis risk and peripheral blood cell counts in patients with essential thrombocythemia (ET), we identified eligible studies from PubMed, Embase, and the Cochrane Library. Seven studies were ultimately included in this meta-analysis. All studies reported the peripheral blood cell counts of ET patients, and three of them reported the eligible thrombotic events. In comparing the effect of MPL versus JAK2V617F on thrombosis, 1257 ET patients (73 MPL + and 1184 JAK2V617F +) were included. MPL-positive (MPL +) ET patients had a higher risk of thrombosis than JAK2V617F-positive (JAK2V617F +) ET patients [RR = 1.80 (1.08–3.01), P = 0.025]. And 3453 ET patients (138 MPL + and 3315 JAK2V617F +) were included in the comparison of peripheral blood cell counts. Platelet counts of MPL + ET patients were higher than that of JAK2V617F + ET patients [WMD = 81.18 (31.77–130.60), P = 0.001]. MPL + ET patients had lower hemoglobin [WMD = − 11.66 (− 14.32 to − 9.00), P = 0.000] and white blood cell counts [WMD = − 1.01 (− 1.47 to − 0.56), P = 0.000] than JAK2V617F + ET patients. These findings indicate that the MPL mutation is a high-risk factor for thrombosis in ET patients, and it may be rational to include MPL mutation in the revised IPSET as a criterion for thrombosis prediction scores. And given the differences in peripheral blood, it is necessary to further study whether MPL + ET patients differ from JAK2V617F + ET patients in bleeding and survival.

Myelofibrosis (MF) Leukemic Transformation (LT) in the Era of Ruxolitinib and Genomics: Characteristics and Outcomes

Introduction: LT of myeloproliferative neoplasms (MPN), including primary or post-polycythemia vera (PV)/essential thrombocythemia (ET) MF, is associated with dismal prognosis, largely owing to the low rates and short duration of responses to currently available therapies. Ruxolitinib is the only approved therapy for MF. No data are available on the characteristics and outcomes of MF LT in pts who received ruxolitinib during the course of MF.Methods: We reviewed our database of consecutive pts with MF followed at our institution from 1993 to 2016 and sorted those who experienced LT. We then divided pts into those who received ruxolitinib during the MF course and those who never did, and compared these two groups in terms of clinical characteristics, biology of LT, response to therapy, and outcome.Results: 135 pts were analyzed: 39 had received ruxolitinib (RUXO), and 96 had not (NO-RUXO). Time to LT was similar between the two groups (45.7 and 45.6 months, P = 0.36), as were median age, gender distribution, proportion of post-PV/ET cases, distribution of driver mutations, or number of treatments for MF (besides ruxolitinib) (all P-values = ns). Similarly, there were no significant differences in peripheral blood and bone marrow parameters of the LT between the two groups, with the possible exception of a lower median Hgb in RUXO pts (8.5 g% vs 9.1 g%, P = 0.05). Complex karyotype was found in 40% and 36% of RUXO and NO-RUXO pts, respectively, and a monosomal karyotype in 20% and 23%, respectively (all P-values = ns). The distribution of relevant mutations, as detected by a previously described next-generation sequencing 28-gene panel (Patel et al. Blood 2015), presented some differences, mostly not statistically significant, likely due to small number of tested pts (Table 1). Notably, however, ASXL1 mutations appeared to be less frequent in RUXO than NO-RUXO pts (12% vs. 63%, respectively P = 0.002). For the purposes of this analysis, induction therapies were grouped into high-dose ara-C (HiDAC)-based, low-dose (Lo)DAC-based, hypomethylating agent (HMA)-based, and others. Similar proportions of patients in the RUXO and NO-RUXO groups received each of these treatment types. Finally, 20% and 25% of the RUXO and NO-RUXO pts, respectively, underwent SCT. The overall response rates (ORR) and complete RR (CRR) were similar between the two groups (40% and 23% for RUXO pts, respectively, and 42% and 38% for NO-RUXO pts, respectively). Similar CRR were achieved in pts who received JAK inhibitors as part of their LT induction regimen and those who did not (20% vs. 37%, P = 0.21). As of this writing, 17 pts are alive. The cause of death was leukemia in the overwhelming majority of pts. The median OS for the entire population, RUXO and NO-RUXO pts is 6.1, 7.9, and 6.1 months, respectively (P = ns). The median PFS is 2.8, 3.1, and 2.6 months, respectively (P = ns). In each group, pts who underwent SCT fared better than those who did not (OS 4.2 vs 18.2 months P < 0.001 and 3.7 vs 18.2 months, respectively, P = 0.004) The presence of ≥3 mutations was independently associated with shorter OS in MF patients treated with ruxolitinib (Patel et al. Blood 2015). We found such association to be of borderline significance in the overall MF LT population (11.7 vs. 1.9 months, P = 0.07) and not statistically significant in the RUXO population (OS 10.1 vs 1.7 months, P = 0.29), in either case likely due to small sample size.Conclusions: RUXO and NO-RUXO pts appeared to have similar responses to induction therapy for MF LT. We did not observe a statistically significantly longer PFS or OS in RUXO pts compared to NO-RUXO pts. Among MF pts who experience LT, prior therapy with ruxolitinib might be associated with a distinct pattern of mutations in the leukemic clone, including lower incidence of ASXL1 mutation. Analysis of a larger sample will be necessary to solidify these findings. [Display omitted] DisclosuresBose:Incyte Corporation: Honoraria. Kantarjian:Novartis: Research Funding; Bristol-Meyers Squibb: Research Funding; Amgen: Research Funding; Delta-Fly Pharma: Research Funding; Pfizer: Research Funding; ARIAD: Research Funding. Verstovsek:Promedior: Research Funding; CTI BioPharma Corp: Research Funding; Pfizer: Research Funding; Gilead: Research Funding; Pfizer: Research Funding; Seattle Genetics: Research Funding; Bristol Myers Squibb: Research Funding; Lilly Oncology: Research Funding; Celgene: Research Funding; Astrazeneca: Research Funding; Celgene: Research Funding; NS Pharma: Research Funding; Roche: Research Funding; Genentech: Research Funding; Gilead: Research Funding; Promedior: Research Funding; Blueprint Medicines Corp: Research Funding; Bristol Myers Squibb: Research Funding; Astrazeneca: Research Funding; Galena BioPharma: Research Funding; Genentech: Research Funding; Blueprint Medicines Corp: Research Funding; Galena BioPharma: Research Funding; CTI BioPharma Corp: Research Funding; Incyte: Research Funding; Incyte: Research Funding; Seattle Genetics: Research Funding; Lilly Oncology: Research Funding; Roche: Research Funding; NS Pharma: Research Funding.

DNA Methylation Differences in Peripheral Blood of Patients with Anaphylaxis.

Objective To study the DNA methylation of nucleated cells in peripheral blood of patients died from anaphylactic shock caused by cephalosporin drugs and to provide a new research direction and basis for the forensic diagnosis of shock caused by drug hypersensitiveness. Methods Methylation microarray was used to detect DNA methylation of nucleated cells in peripheral blood of patients died from anaphylactic shock caused by cephalosporin drugs and normal subjects. Sequencing data and chip data were analyzed for differences in DNA methylation using R language methylkit, ChAMP package. Random forest algorithm was used to evaluate the importance of the DNA methylation differential sites. Results Differential sites of DNA methylation highly associated with anaphylaxis caused by cephalosporin drugs were obtained at loci such as ETS1, PRR23B and GNAS. Conclusion Cephalosporin allergy is associated with DNA methylation, and DNA methylation may be a new strategy for forensic identification of anaphylactic shock and death.

Cell Ratio Differences in Peripheral Blood between Early- and Late-Onset Parkinson's Disease: A Case-Control Study.

Objectives To explore the differences of immune disorders in peripheral blood between patients with early-onset Parkinson's disease (EOPD) and late-onset Parkinson's disease (LOPD). Methods We retrospectively reviewed medical records of Parkinson's disease (PD) patients and healthy controls between June 2002 and July 2017. At last, we included 117 PD patients who were divided into EOPD and LOPD according to whether onset age of PD was after 50 and 99 controls divided into E-Control (match for EOPD) and L-Control (match for LOPD) according to whether their age was after 53 which was onset age plus median of disease duration. We compared the ratios of cells between multiple groups and performed the multinominal logistic regression analysis to explore the relationship between ratios and subtypes of PD. We also carried out the receiver operating characteristic (ROC) curve analysis to estimate the diagnostic value of the variable. Results Lymphocyte-red blood cell ratio (LRR) was lower in LOPD compared with that in EOPD or L-Control. LRR was also negatively associated with LOPD (OR: 0.623; 95% CI: 0.397–0.980; P=0.040). The ROC curve analysis showed the optimal cutoff value of 4.53 (×10−4) of LRR for discrimination of LOPD versus L-Control (sensitivity: 0.596, specificity: 0.764). The area under curve (AUC) was 0.721. As for LOPD versus EOPD, the optimal threshold of LRR was 4.10 (×10−4) (sensitivity: 0.516, specificity: 0.745). AUC was 0.641. Conclusions Peripheral immune disorders might play an important part in the pathological progression of LOPD. Also, LRR has potential diagnostic value.

Symptom-based stratification of patients with primary Sjögren's syndrome: multi-dimensional characterisation of international observational cohorts and reanalyses of randomised clinical trials.

SummaryBackgroundHeterogeneity is a major obstacle to developing effective treatments for patients with primary Sjögren's syndrome. We aimed to develop a robust method for stratification, exploiting heterogeneity in patient-reported symptoms, and to relate these differences to pathobiology and therapeutic response.MethodsWe did hierarchical cluster analysis using five common symptoms associated with primary Sjögren's syndrome (pain, fatigue, dryness, anxiety, and depression), followed by multinomial logistic regression to identify subgroups in the UK Primary Sjögren's Syndrome Registry (UKPSSR). We assessed clinical and biological differences between these subgroups, including transcriptional differences in peripheral blood. Patients from two independent validation cohorts in Norway and France were used to confirm patient stratification. Data from two phase 3 clinical trials were similarly stratified to assess the differences between subgroups in treatment response to hydroxychloroquine and rituximab.FindingsIn the UKPSSR cohort (n=608), we identified four subgroups: Low symptom burden (LSB), high symptom burden (HSB), dryness dominant with fatigue (DDF), and pain dominant with fatigue (PDF). Significant differences in peripheral blood lymphocyte counts, anti-SSA and anti-SSB antibody positivity, as well as serum IgG, κ-free light chain, β2-microglobulin, and CXCL13 concentrations were observed between these subgroups, along with differentially expressed transcriptomic modules in peripheral blood. Similar findings were observed in the independent validation cohorts (n=396). Reanalysis of trial data stratifying patients into these subgroups suggested a treatment effect with hydroxychloroquine in the HSB subgroup and with rituximab in the DDF subgroup compared with placebo.InterpretationStratification on the basis of patient-reported symptoms of patients with primary Sjögren's syndrome revealed distinct pathobiological endotypes with distinct responses to immunomodulatory treatments. Our data have important implications for clinical management, trial design, and therapeutic development. Similar stratification approaches might be useful for patients with other chronic immune-mediated diseases.FundingUK Medical Research Council, British Sjogren's Syndrome Association, French Ministry of Health, Arthritis Research UK, Foundation for Research in Rheumatology.Video Symptom-based stratification of patients with Sjögren's syndrome

Distribution and functions of \u03b3\u03b4 T cells infiltrated in the ovarian cancer microenvironment

BackgroundThe role of γδ T cells, innate-like lymphocytes with unrestrained MHC, in various malignancies has recently been extensively studied. However, there is limited research regarding γδ T cells in ovarian cancer (OC) patients.MethodsHere, we investigated the distribution patterns of γδ T cells and their main subsets in peripheral blood and tumor tissues among OC patients, benign ovarian tumor (BOT) patients, and age-matched healthy controls (HC) by flow cytometry, as well as the expression levels of IFN-γ and IL-17A secreted from γδ T cells. Immunohistochemical staining was utilized to detect the numbers of γδ T cells and their main subsets in different types of ovarian tumor tissues. Additionally, we also investigated chemotaxis effects on γδ T cells, as well as their cytotoxic activity and proliferation.ResultsWe found that the percentages of γδ T cells and Vδ1 T cells were significantly higher in OC tissues than BOT tissues and normal (N) ovarian tissues, while there were no obvious differences in peripheral blood. Meanwhile, higher numbers of γδ T cells and Vδ1 T cells were observed in OC tissues, and were positively related to advanced clinicopathological features of OC patients. Further, the levels of IFN-γ secreted by γδ T cells were relatively lower, while IL-17A was expressed at a high level in both the peripheral blood and tissues of OC patients. Chemotaxis assay revealed that supernatants derived from OC tissues possessed a stronger capacity to attract and recruit γδ T cells. However, γδ T cells sorted from OC tissues showed weakened cytotoxic activity against ovarian cancer cells, and γδ T cells cocultured with OC tissue supernatants could effectively inhibit the proliferative activity of naïve CD4+ T cells.ConclusionsThese data suggested that γδ T cells might have critical roles in OC progression and potential utilization in treatment approaches or prognosis prediction.

Exposure to polybrominated biphenyl (PBB) associates with genome-wide DNA methylation differences in peripheral blood

ABSTRACTIn 1973, Michigan residents were exposed to polybrominated biphenyl (PBB) when it was accidentally added to farm animal feed. Highly exposed individuals and their children have experienced endocrine-related health problems, though the underlying mechanism behind these remains unknown. We investigated whether PBB exposure is associated with variation in DNA methylation in peripheral blood samples from 658 participants of the Michigan PBB registry using the MethylationEPIC BeadChip, as well as investigated what the potential function of the affected regions are and whether these epigenetic marks are known to associate with endocrine system pathways. After multiple test correction (FDR <0.05), 1890 CpG sites associated with total PBB levels. These CpGs were not enriched in any particular biological pathway, but were enriched in enhancer and insulator regions, and depleted in regions near the transcription start site or in CpG islands (p < 0.05). They were also more likely to be in ARNT and ESR2 transcription factor binding sites (p = 3.27e-23 and p = 1.62e-6, respectively), and there was significant overlap between CpGs associated with PBB and CpGs associated with estrogen (p < 2.2e-16). PBB-associated CpGs were also enriched for CpGs known to be associated with gene expression in blood (eQTMs) (p < 0.05). These eQTMs were enriched for pathways related to immune function and endocrine-related autoimmune disease (FDR <0.05). These results indicate that exposure to PBB is associated with differences in epigenetic marks that suggest that it is acting similarly to estrogen and is associated with dysregulated immune system pathways.

Diagnostic Value of Peripheral and Bronchoalveolar Leukocyte Profile in Lung Transplant Recipients After Alemtuzumab Induction Therapy. A Single Center Experience

Early diagnosis of lower respiratory tract infection (LRTI) and acute cellular rejection (ACR) is crucial for adequate treatment in lung transplant recipients. A recently described algorithm (Speck 2016) for LRTI and ACR prediction which includes differences in peripheral blood (PBIC) and bronchoalveolar lavage immune cells (BALIC) was tested in LuTX patients treated with alemtuzumab induction therapy.

\u201cDNA Methylation signatures in panic disorder\u201d

Panic disorder (PD) affects about four million Europeans, with women affected twice as likely as men, causing substantial suffering and high economic costs. The etiopathogenesis of PD remains largely unknown, but both genetic and environmental factors contribute to risk. An epigenome-wide association study (EWAS) was conducted to compare medication-free PD patients (n = 89) with healthy controls (n = 76) stratified by gender. Replication was sought in an independent sample (131 cases, 169 controls) and functional analyses were conducted in a third sample (N = 71). DNA methylation was assessed in whole blood using the Infinium HumanMethylation450 BeadChip. One genome-wide association surviving FDR of 5% (cg07308824, P = 1.094 × 10-7, P-adj = 0.046) was identified in female PD patients (N = 49) compared to controls (N = 48). The same locus, located in an enhancer region of the HECA gene, was also hypermethylated in female PD patients in the replication sample (P = 0.035) and the significance of the association improved in the meta-analysis (P-adj = 0.004). Methylation at this CpG site was associated with HECA mRNA expression in another independent female sample (N = 71) both at baseline (P = 0.046) and after induction by dexamethasone (P = 0.029). Of 15 candidates, 5 previously reported as associated with PD or anxiety traits also showed differences in DNA methylation after gene-wise correction and included SGK1, FHIT, ADCYAP1, HTR1A, HTR2A. Our study examines epigenome-wide differences in peripheral blood for PD patients. Our results point to possible sex-specific methylation changes in the HECA gene for PD but overall highlight that this disorder is not associated with extensive changes in DNA methylation in peripheral blood.

T24 - Sex-Specific Dna Methylation Signatures In Panic Disorder

Background Panic disorder (PD) is characterized by sudden episodes of acute anxiety occurring without any apparent reason. PD is the most disabling anxiety disorder and it affects twice as many women as men. The heritability of PD is estimated to be up to 48% and epidemiological studies show that both cumulative and specific life events are risk factors for the development of PD. Therefore, the investigation of genetic factors and gene-environment interaction is of high importance for understanding the pathophysiology of PD. As such, examining epigenetic differences in PD patients is of great interest given that environmental factors, in combination with genetic variation, can influence DNA methylation. Methods We conducted an Epigenome-Wide Association Study (EWAS) comparing non-medicated PD patients (49 females, 40 males) with healthy controls (48 females, 28 males). Replication was sought in an independent sample (131 cases, 190 controls) and further confirmed with a meta-analysis across both samples (220 cases, 266 controls). DNA methylation levels were assessed in whole blood using the Infinium HumanMethylation450 BeadChip. Failed probes were excluded based on a detection p-value larger than 0.01 in >50% of the samples. X chromosome, Y chromosome and non-specific binding probes were removed. The data were normalized with functional normalization in Minfi and batch-corrected using ComBat. Regression analyses accounting for cellular heterogeneity, sex and age were performed to test for associations between PD status and DNA methylation. To further assess functionality of the significant CpG, gene expression profiles (Illumina HumanHT-12 v3.0 array) in peripheral blood before and after exposure to the glucocorticoid receptor agonist dexamethasone were tested for association with DNA methylation in another independent female sample (N=71). Results No genome-wide associations were observed in PD patients compared to controls in the whole sample. Interestingly when stratified by sex, only the comparison of female PD patients with controls yielded one genome-wide association surviving FDR of 5% (P= 1.094 x 10-7 , P-adj=0.046). Specifically, cg07308824, located in the promoter of the HECA gene, was hypermethylated in female PD patients (N=49) compared to controls (N=48). The same CpG was also hypermethylated in female PD patients in the replication sample (P=0.035) and the genome-wide significant association was confirmed in the meta-analysis (P-adj=0.007). Methylation at this CpG site was associated with mRNA expression of HECA both at baseline (P= 0.046) and after induction by dexamethasone (P= 0.029). Pathway analyses using Web Gestalt investigating the top 50 associated CpG sites showed enrichment in female-specific pathways, e.g. female infertility pathways. Discussion Our study is the first to examine epigenome wide differences in peripheral blood for PD patients. Interestingly, our results point to possible sex-specific and functional methylation changes in PD. Further studies are needed to examine a possible contribution of the HECA gene methylation to female specific factors in PD.

Gene Expression Differences in Peripheral Blood of Parkinson's Disease Patients with Distinct Progression Profiles.

The prognosis of neurodegenerative disorders is clinically challenging due to the inexistence of established biomarkers for predicting disease progression. Here, we performed an exploratory cross-sectional, case-control study aimed at determining whether gene expression differences in peripheral blood may be used as a signature of Parkinson’s disease (PD) progression, thereby shedding light into potential molecular mechanisms underlying disease development. We compared transcriptional profiles in the blood from 34 PD patients who developed postural instability within ten years with those of 33 patients who did not develop postural instability within this time frame. Our study identified >200 differentially expressed genes between the two groups. The expression of several of the genes identified was previously found deregulated in animal models of PD and in PD patients. Relevant genes were selected for validation by real-time PCR in a subset of patients. The genes validated were linked to nucleic acid metabolism, mitochondria, immune response and intracellular-transport. Interestingly, we also found deregulation of these genes in a dopaminergic cell model of PD, a simple paradigm that can now be used to further dissect the role of these molecular players on dopaminergic cell loss. Altogether, our study provides preliminary evidence that expression changes in specific groups of genes and pathways, detected in peripheral blood samples, may be correlated with differential PD progression. Our exploratory study suggests that peripheral gene expression profiling may prove valuable for assisting in prediction of PD prognosis, and identifies novel culprits possibly involved in dopaminergic cell death. Given the exploratory nature of our study, further investigations using independent, well-characterized cohorts will be essential in order to validate our candidates as predictors of PD prognosis and to definitively confirm the value of gene expression analysis in aiding patient stratification and therapeutic intervention.

Patients With a History of Chemotherapy and Isolated del(20q) With Minimal Myelodysplasia Have an Indolent Course

Isolated deletion (20q) is relatively common in myeloid neoplasms and has been rarely reported in cases of therapy-related myelodysplastic syndrome (MDS). Our aim was to characterize cases of isolated del(20q) in bone marrow biopsy specimens from patients with a history of chemotherapy with morphologic findings insufficient for a diagnosis of MDS. In this retrospective study from one institution, we identified 22 patients with isolated del(20q) and no or minimal dysplasia and evaluated clinical and pathologic characteristics. Eleven of the patients had a history of chemotherapy for mostly lymphoproliferative disorders. There were no statistically significant differences in peripheral blood or bone marrow features between patients with a history of chemotherapy and those without. Three patients with a history of chemotherapy had died at last follow-up; cause of death was recurrent nonmyeloid neoplasm. None of the patients with a history of chemotherapy subsequently developed a high-grade myeloid neoplasm, whereas one of the patients who had not received prior chemotherapy developed refractory anemia with excess blasts 2. The presence of del(20q) as an isolated bone marrow cytogenetic abnormality in the absence of morphologic findings sufficient for a diagnosis of acute myeloid leukemia, myeloproliferative neoplasm, or MDS portends an indolent clinical course, regardless of previous exposure to chemotherapy.

LOCAL AND SYSTEMIC IMMUNE PARAMETERS IN SEVERE ATOPIC DERMATITIS AND CUTANEOUS T-CELL LYMPHOMA PATIENTS

Background.. To investigate expression of cytokines genes parameters in skin and blood in severe atopic dermatitis (AD) and cutaneous T-cell lymphoma (CTCL) patients comparing with healthy donors. Materials and methods. 20 severe AD patients, 20 CTCL patients and 20 healthy donors were included in the study. Skin samples and peripheral blood were used as material for immunological study. Interleukins — (IL)1B, IL2, IL2r, IL4, IL5, IL6, IL7, IL8, IL10, IL12A, IL12B, IL15 (total), IL15 , IL17A, IL18, IL23, IL28, IL29, Interferon γ (IFNγ), tumor necrosis factor α (TNFα), transforming growth factor beta 1 (TGFB1), forkhead box P3 (FOXP3) gene expression was defined in the skin and peripheral blood of severe AD patients, CTCL patients and healthy donors by real-time reverse transcription polymerase chain reaction (RT-PCR). Results. Statistically significant increase of cytokines genes IL4,IL12B,IL17A, TNFα in peripheral blood of severe AD patients compared with CTCL patients was marked. Studying of skin samples from CTCL patients has shown statistically significant increase of cytokines IL8, IL10, IL15, IFNγ genes expression and decrease of IL18 gene expression in comparison with skin samples from severe AD patients. Conclusion. Obtained cytokines genes expression cytokines genes parameters in peripheral blood of severe AD and CTCL patients had a certain similarities consisting of increased IL8, IFNγ and decreased IL6, IL23 genes expression in comparison with healthy donors. Substantial differences in peripheral blood of patients in comparison with healthy donors: increased IL-5, IL-12B genes expression in AD patients and decreased IL-8, IL-12A genes expression in CTCL patients, at the same time increased expression of IL-6, IL-8, IL-10, IFNγ genes in severe AD and CTCL patients were shown.

Gene-expression differences in peripheral blood between lithium responders and non-responders in the Lithium Treatment-Moderate dose Use Study (LiTMUS)

This study was designed to identify genes whose expression in peripheral blood may serve as early markers for treatment response to lithium (Li) in patients with bipolar disorder. Although changes in peripheral blood gene-expression may not relate directly to mood symptoms, differences in treatment response at the biochemical level may underlie some of the heterogeneity in clinical response to Li. Subjects were randomized to treatment with (n=28) or without (n=32) Li. Peripheral blood gene-expression was measured before and 1 month after treatment initiation, and treatment response was assessed after 6 months. In subjects treated with Li, 62 genes were differentially regulated in treatment responders and non-responders. Of these, BCL2L1 showed the greatest difference between Li responders and non-responders. These changes were specific to Li responders (n=9), and were not seen in Li non-responders or patients treated without Li, suggesting that they may have specific roles in treatment response to Li.

Hormonal Differences in Peripheral Blood and Gene Profiling in the Liver and Lymphocytes in Japanese Black Cattle with Growth Retardation

Japanese Black cattle occasionally demonstrate growth retardation despite sufficient nutrient intake. To clarify hormonal and transcriptional characteristics, we investigated differences in blood components, including hormones, and differences in exhaustive gene expressions in the liver and peripheral lymphocytes of six cattle with growth retardation (GR cattle) and eight control cattle of the same age and pedigree with normal growth. Hematocrit values and concentrations of hemoglobin, serum albumin, total cholesterol, insulin-like growth factor 1 (IGF-1), thyroxine and insulin in GR cattle were significantly lower than those in controls. GR cattle also excreted higher levels of GH. We used three GR and three control cattle for a microarray analysis in the liver and found that 279 gene expressions were significantly different. However, gene expressions related to the GH-IGF-1 axis, such as the GH receptor and IGF-1, were not significantly different from those of controls. Immune-related gene expressions were significantly lower. To clarify these gene expression levels, peripheral lymphocytes were used for real-time RT-PCR. The expression rates of genes that were significantly lower in the liver, such as chemokine ligand 8, interferon gamma receptor 1 and immunoglobulin light chain VJ region were also significantly lower in three GR cattle than those in the three control cattle. These results suggest that the cause of growth retardation in the present study was due to other factors, not abnormal gene expressions of factors related to the GH-IGF-1 axis in the liver, and that GR cattle were susceptible to infectious disease.

Nrf2 Deficiency Leads to Altered Hematopoietic Stem Cell Function and Increased Sensitivity to Alkylating Agent Induced Myeloid Dysplasia,

Abstract 3828Therapy-related myelodysplasia or acute myeloid leukemia (t-MDS/AML) is a lethal complication of cancer treatment. In a previous study we conducted gene expression analysis comparing peripheral blood stem cell (PBSC) CD34+ cells from patients who subsequently developed t-MDS/AML after autologous hematopoietic cell transplantation (aHCT) for Hodgkin lymphoma (HL) or non-Hodgkin lymphoma (NHL) and matched controls that did not develop t-MDS/AML after aHCT. This study showed that the Nrf2-mediated oxidative stress response was one of the top ranked canonical pathways downregulated in t-MDS/AML cases compared to controls. Nrf2 is a basic region-leucine-zipper transcription factor that regulates expression of numerous detoxifying and antioxidant genes as well as ubiquitination and proteasomal degradation proteins. Since the Nrf2 pathway is a major cellular defense mechanism against oxidative and xenobiotic stresses, we hypothesized that downregulated Nrf2 response may cause increased toxicity after genotoxic therapeutic exposures and contribute to development of t-MDS/AML. To test this hypothesis, we studied Nrf2 knockout mice to determine the effects of Nrf2 deficiency on hematopoiesis under physiological conditions and after exposure to genotoxic stress. We did not observe significant differences in peripheral blood (PB) counts between Nrf2 knockout (KO) mice and wild-type (WT) mice. Analyses of hematopoietic cells from PB, bone marrow (BM) or spleen using multi-color flow cytometry demonstrated that the compositions of granulocyte, B cell and T cell lineages were similar between KO and WT mice. No differences in BM long-term hematopoietic stem cell (LT-HSC, Lin-c-Kit+Sca-1+ Flt3- CD48-CD150+), multi-potent progenitor (MPP, Lin-c-Kit+Sca-1+ Flt3- CD48+), common-myeloid progenitor (CMP, Lin-c-Kit+Sca-1-CD34+CD16/32-), granulocyte/monocyte progenitor (GMP, Lin-c-Kit+Sca-1-CD34+CD16/32+), or megakaryocyte/erythroid progenitor (MEP, Lin-c-Kit+Sca-1-CD34-CD16/32-) populations were seen between KO and WT mice. However, analysis of LT-HSC function using competitive repopulation assays demonstrated significantly reduced chimerism of donor derived cells in PB of recipient mice at 12 weeks and 16 weeks in KO group compared to WT control (42.49±3.09% vs. 54.75±2.13% at 12 weeks and 42.5±2.8% vs. 57.2±3.3% at 16 weeks for KO and WT respectively). Although we did not see significant differences in chimerism of donor-derived hematopoietic stem/progenitor cells in the BM of KO and WT groups at 16 weeks, we did observe significantly reduced contribution of donor-derived cells from the KO group compared to WT control in PB in secondary recipients as early as 8 weeks after secondary transplantation of BM cells (16.5±1.7% vs 61.9±1.5% for KO and WT respectively). These data indicate that Nrf2 deficiency causes cell-intrinsic functional defects in HSC self-renewal. In order to test if Nrf2 deficient hematopoietic cells are more sensitive to genotoxic stress compared to WT cells, we treated KO and WT mice with the alkylating agent ENU and monitored for hematological changes in PB. We did not observe significant differences in hematopoietic recovery after exposure to acute genotoxic stress between KO and WT mice. However, KO mice developed severe anemia at a significantly increased rate and with shorter latency compared to WT mice after ENU treatment (88% in KO vs. 28% in WT at 4 months, p=0.003). Anemic mice developed massive splenomegaly, and histological analysis showed loss of follicular structure and dramatic increase in erythropoiesis with areas of myeloid elements. Pathological examination of BM from anemic mice revealed normal cellularity but reduced erythropoiesis with expansion of myeloid cells. FACS analyses revealed expansion of myeloid (CD11b+, Gr-1+) and immature erythroid population (CD71+Ter119+/dim) in the spleen and BM, and increased CD11b+, Gr-1+ cells in PB. These findings are consistent with development of myeloid dysplasia in these mice. In conclusion, Nrf2 deficiency results in alterations in HSC self-renewal capacity under physiological condition, and leads to increased sensitivity to alkylator-induced myeloid dysplasia in mice. Disclosures:No relevant conflicts of interest to declare.