ABSTRACT
OBJECTIVE: To quantitatively assess the concentration and distribution of body iron(BI) in children aged 6 to 17 years in Beijing. METHODS: A total of 1392 children aged 6 to 17 years in Beijing were randomly selected for questionnaire survey and physical examination in 2016-2017. Fasting venous blood was collected, serum ferritin(SF) and serum soluble transferrin receptor(sTfR) were measured by immunoturbidimetric assay. BI were calculated, and the concentration and distribution of BI in children aged 6 to 17 years was assessed. RESULTS: The average BI level of children aged 6 to 17 in Beijing was(5.49±2.94) mg/kg. The average BI level of boys was higher than that of girls, and the average BI level of children with abdominal obesity was higher than that of children without abdominal obesity, and the differences were statistically significant(P<0.05). The concentration of BI in children aged 6 to 17 years in Beijing was 4-7.99 mg/kg, accounting for the highest proportion(57.3%). The concentration of BI<0 mg/kg accounted for the lowest proportion(4.3%). There were significant differences in the distribution of BI concentration in different gender, age and abdominal obesity(P<0.05). The anemia rate of children aged 6 to 17 in Beijing was 2.7%(95%CI 1.9-3.6), the low ferritin rate(SF<25 µg/L) was 22.5%(95%CI 20.0-24.8), and the iron deficiency rate(SF<15 µg/L) was 7.8%(95%CI 6.4-9.3), the negative iron stores(BI<0 mg/kg) rate was 4.3%(95%CI 3.2-5.3). The anemia rate, low ferritin rate, iron deficiency rate and negative iron stores rate were higher in girls than boys, and higher in children aged 12 to 17 years than in children aged 6 to 11 years, and the differences were statistically significant(P<0.01). CONCLUSION: Iron deficiency was still present in children aged 6 to 17 in Beijing from 2016 to 2017. The proportion of low BI level in girls and children aged 12 to 17 is higher, respectively.
Subject(s)
Anemia, Iron-Deficiency , Anemia , Iron Deficiencies , Male , Child , Female , Humans , Ferritins , Anemia, Iron-Deficiency/epidemiology , Beijing , Obesity, Abdominal , Iron , Receptors, Transferrin , ObesityABSTRACT
OBJECTIVE: To evaluate the nutritional status of vitamin D in school-age children in Beijing. METHODS: The data was part of the monitoring data of Beijing in "Chinese Nutrition and Health Monitoring of Children and Nursing Mothers 2016-2017". A total of 1385 students were recruited from 10 primary schools, 10 junior middle schools and 5 senior high schools. The concentrations of serum vitamin D were determined using electrochemiluminescence immunoassay. The distribution of serum vitamin D in school-age children was further described for different regions, age, body mass index(BMI), waistline, outdoor activity time and myopia. The prevalence of the insufficiency and deficiency of vitamin D was compared in different subgroups. RESULTS: The median concentration of serum 25(OH)D_3 [M(P25, P75)] of students aged 6-11 and 12-17 were 20.86(16.48, 25.31) ng/mL and 14.12(11.04, 19.17) ng/mL. The serum 25(OH)D_3 of male school-age children was higher than that of female. The serum 25(OH)D_3 of students aged 12-17 was lower than that of students aged 6-11. The serum 25(OH)D_3 of urban students was lower than that of rural students, and the serum 25(OH)D_3 of myopia students was lower than that of non-myopia students(P<0.01). There was no significant difference in serum 25(OH)D_3 between students with outdoor activity time greater than or equal to 2 hours and students with less than 2 hours, normal weight and overweight and obese students, normal waist and abdominal obesity students. The vitamin D deficiency rate, insufficiency rate, insufficiency and deficiency rate of school-age children were 18.8%(261), 40.1%(556) and 59.0%(817). The vitamin D deficiency in girls was more serious than that in boys, and the vitamin D insufficiency and deficiency of students aged 12-17 were more serious than that of students aged 6-11. The vitamin D deficiency of students in urban areas was more serious than that in rural areas, and the vitamin D deficiency of myopia students was more serious than that of non-myopia(P<0.01). There was no significant difference in vitamin D deficiency between students with outdoor activity time greater than or equal to 2 hours and students with less than 2 hours, normal weight and overweight and obese students, normal waist and abdominal obesity students. The statistically significant variables(gender, age, region, outdoor activity time, overweight, obesity, abdominal obesity and myopia) were included in the multivariate Logistic regression model for analysis. The result showed that girls(OR = 2.005, P<0.001), 12-17 years old(OR=4.310, P<0.001), rural(OR=0.586, P<0.001) with vitamin D deficiency and insufficiency were more likely. CONCLUSION: The vitamin D status of students in Beijing is not ideal, and the deficiency and insufficiency of vitamin D in girls is more serious than that in boys; The deficiency and insufficiency of vitamin D in 12-17 years old is more serious than that in 6-11 years old; The vitamin D deficiency of urban students is more serious than that of rural students.
Subject(s)
Overweight , Vitamin D Deficiency , Humans , Male , Child , Female , Adolescent , Overweight/epidemiology , Vitamin D , Beijing/epidemiology , Obesity, Abdominal , Urban Population , Obesity/epidemiology , Body Mass Index , Vitamin D Deficiency/epidemiology , Vitamins , PrevalenceABSTRACT
Small cell lung cancer (SCLC) is an aggressive disease with poor survival. A few sequencing studies performed on limited number of samples have revealed potential disease-driving genes in SCLC, however, much still remains unknown, particularly in the Asian patient population. Here we conducted whole exome sequencing (WES) and transcriptomic sequencing of primary tumors from 99 Chinese SCLC patients. Dysregulation of tumor suppressor genes TP53 and RB1 was observed in 82% and 62% of SCLC patients, respectively, and more than half of the SCLC patients (62%) harbored TP53 and RB1 mutation and/or copy number loss. Additionally, Serine/Arginine Splicing Factor 1 (SRSF1) DNA copy number gain and mRNA over-expression was strongly associated with poor survival using both discovery and validation patient cohorts. Functional studies in vitro and in vivo demonstrate that SRSF1 is important for tumorigenicity of SCLC and may play a key role in DNA repair and chemo-sensitivity. These results strongly support SRSF1 as a prognostic biomarker in SCLC and provide a rationale for personalized therapy in SCLC.
Subject(s)
Carcinoma, Small Cell/genetics , Lung Neoplasms/genetics , Oncogene Proteins/genetics , Serine-Arginine Splicing Factors/genetics , Adult , Aged , DNA Copy Number Variations , DNA Damage , Female , Gene Silencing , Humans , Male , Middle Aged , MutationABSTRACT
Genome-wide association studies have successfully identified a subset of common variants associated with lung cancer risk. However, these variants explain only a fraction of lung cancer heritability. It has been proposed that low-frequency or rare variants might have strong effects and contribute to the missing heritability. To assess the role of low-frequency or rare variants in lung cancer development, we analyzed exome chips representing 1,348 lung cancer subjects and 1,998 control subjects during the discovery stage and subsequently evaluated promising associations in an additional 4,699 affected subjects and 4,915 control subjects during the replication stages. Single-variant and gene-based analyses were carried out for coding variants with a minor allele frequency less than 0.05. We identified three low-frequency missense variants in BAT2 (rs9469031, c.1544C>T [p.Pro515Leu]; odds ratio [OR] = 0.55, p = 1.28 × 10(-10)), FKBPL (rs200847762, c.410C>T [p.Pro137Leu]; OR = 0.25, p = 9.79 × 10(-12)), and BPIFB1 (rs6141383, c.850G>A [p.Val284Met]; OR = 1.72, p = 1.79 × 10(-7)); these variants were associated with lung cancer risk. rs9469031 in BAT2 and rs6141383 in BPIFB1 were also associated with the age of onset of lung cancer (p = 0.001 and 0.006, respectively). BAT2 and FKBPL at 6p21.33 and BPIFB1 at 20q11.21 were differentially expressed in lung tumors and paired normal tissues. Gene-based analysis revealed that FKBPL, in which two independent variants were identified, might account for the association with lung cancer risk at 6p21.33. Our results highlight the important role low-frequency variants play in lung cancer susceptibility and indicate that candidate genes at 6p21.33 and 20q11.21 are potentially biologically relevant to lung carcinogenesis.
Subject(s)
Genetic Predisposition to Disease , Genome-Wide Association Study , Lung Neoplasms/genetics , Asian People , Autoantigens/genetics , Chromosomes, Human, Pair 20/genetics , Chromosomes, Human, Pair 6/genetics , Fatty Acid-Binding Proteins , Female , Gene Frequency , Genotype , Humans , Immunophilins/genetics , Lung Neoplasms/pathology , Male , Proteins/genetics , Risk Factors , Tacrolimus Binding ProteinsABSTRACT
Type I IFNs play a critical role in the immune response to viral infection and may also drive autoimmunity through modulation of monocyte maturation and promotion of autoreactive lymphocyte survival. Recent demonstrations of type I IFN gene signatures in autoimmune diseases, including scleroderma, led us to investigate the pathological role of IFNs in a preclinical model of sclerodermatous graft-versus-host disease. Using a neutralizing Ab against the type I IFN receptor IFNAR1, we observed a marked reduction in dermal inflammation, vasculopathy, and fibrosis compared with that seen in the presence of intact IFNAR1 signaling. The ameliorative effects of IFNAR1 blockade were restricted to the skin and were highly associated with inhibition of chronic vascular injury responses and not due to the inhibition of the T or B cell alloresponse. Inhibition of IFNAR1 normalized the overexpression of IFN-inducible genes in graft-versus-host disease skin and markedly reduced dermal IFN-α levels. Depletion of plasmacytoid dendritic cells, a major cellular source of type I IFNs, did not reduce the severity of fibrosis or type I IFN gene signature in the skin. Taken together, these studies demonstrate an important role for type I IFN in skin fibrosis, and they provide a rationale for IFNAR1 inhibition in scleroderma.
Subject(s)
Dendritic Cells/immunology , Graft vs Host Disease/immunology , Inflammation/immunology , Interferon-alpha/metabolism , Scleroderma, Systemic/immunology , Skin/pathology , Vascular Diseases/immunology , Animals , Antibodies, Blocking/administration & dosage , Autoantibodies/blood , Disease Models, Animal , Female , Fibrosis , Gene Expression Regulation , Humans , Interferon-alpha/genetics , Mice , Mice, Inbred C57BL , Mice, Transgenic , Receptor, Interferon alpha-beta/immunology , Signal TransductionABSTRACT
Tumors are characterized by properties of genetic instability, heterogeneity, and significant oligoclonality. Elucidating this intratumoral heterogeneity is challenging but important. In this study, we propose a framework, BubbleTree, to characterize the tumor clonality using next generation sequencing (NGS) data. BubbleTree simultaneously elucidates the complexity of a tumor biopsy, estimating cancerous cell purity, tumor ploidy, allele-specific copy number, and clonality and represents this in an intuitive graph. We further developed a three-step heuristic method to automate the interpretation of the BubbleTree graph, using a divide-and-conquer strategy. In this study, we demonstrated the performance of BubbleTree with comparisons to similar commonly used tools such as THetA2, ABSOLUTE, AbsCN-seq and ASCAT, using both simulated and patient-derived data. BubbleTree outperformed these tools, particularly in identifying tumor subclonal populations and polyploidy. We further demonstrated BubbleTree's utility in tracking clonality changes from patients' primary to metastatic tumor and dating somatic single nucleotide and copy number variants along the tumor clonal evolution. Overall, the BubbleTree graph and corresponding model is a powerful approach to provide a comprehensive spectrum of the heterogeneous tumor karyotype in human tumors. BubbleTree is R-based and freely available to the research community (https://www.bioconductor.org/packages/release/bioc/html/BubbleTree.html).
Subject(s)
Aneuploidy , High-Throughput Nucleotide Sequencing/methods , Neoplasms/genetics , Software , Algorithms , DNA Copy Number Variations , Humans , Sequence Analysis, DNA/methodsABSTRACT
Preferential pathways can be significant vapor intrusion (VI) contributors, causing potentially higher inhalation risk to residents of affected buildings than that arising through traditional intrusion pathways. To assess land drains as a preferential pathway, a three-dimensional model, validated using data from a 4-yr field study, was used to study the roles of subfoundation soil permeability on soil gas flow and indoor depressurization. Results indicated that it is almost impossible for an indirect preferential pathway like a land drain ending in subfoundation soils with a permeability <10 m to affect indoor air quality if the land drain connects to a source with the same vapor concentration as that of the groundwater source beneath the building. An equation was developed to estimate the threshold permeability. We also found that even after the preferential pathway was identified using indoor depressurization (also known as controlled pressure method [CPM]) and then turned off, the influence of the preferential pathway and indoor depressurization on indoor concentration might last for months, although it may not be significant (i.e., may not exceed one order of magnitude, in this study). In the absence of such a preferential VI pathway, CPM may actually reduce indoor air concentrations of contaminants below those present under natural indoor pressure conditions, due to the emission rate limit determined by the upward diffusion rate from the vapor source. Our study highlights the role of measuring subfoundation soil permeability to soil gas flow in site investigations and warns practitioners about the possible mischaracterization of indoor air concentration after applying CPM in the absence of a preferential pathway.
Subject(s)
Air Pollution, Indoor/analysis , Gases/analysis , Groundwater , Soil/chemistryABSTRACT
Long noncoding RNAs (lncRNAs) have recently been identified to be tightly linked to diverse human diseases. However, our knowledge of Systemic Lupus Erythematosus (SLE)-related lncRNAs remains limited. In the present study we investigated the contribution of the lncRNA NEAT1 to the pathogenesis of SLE. Here, we found NEAT1 expression was abnormally increased in SLE patients and predominantly expressed in human monocytes. Additionally, NEAT1 expression was induced by LPS via p38 activation. Silencing NEAT1 significantly reduced the expression of a group of chemokines and cytokines, including IL-6, CXCL10, etc., which were induced by LPS continuously and in late stages. Furthermore, it was identified the involvement of NEAT1 in TLR4-mediated inflammatory process was through affecting the activation of the late MAPK signaling pathway. Importantly, there was a positive correlation between NEAT1 and clinical disease activity in SLE patients. In conclusion, the increased NEAT1 expression may be a potential contributor to the elevated production of a number of cytokines and chemokines in SLE patients. Our findings suggest lncRNA contributes to the pathogenesis of lupus and provides potentially novel target for therapeutic intervention.
Subject(s)
Cytokines/immunology , Inflammation Mediators/immunology , Lupus Erythematosus, Systemic/immunology , MAP Kinase Signaling System/immunology , RNA, Long Noncoding/immunology , Blotting, Western , Cell Line, Tumor , Cluster Analysis , Cytokines/genetics , Cytokines/metabolism , Gene Expression Profiling/methods , Gene Expression Regulation/drug effects , Gene Expression Regulation/immunology , Gene Ontology , Humans , Inflammation Mediators/metabolism , Lipopolysaccharides/pharmacology , Lupus Erythematosus, Systemic/genetics , Lupus Erythematosus, Systemic/metabolism , MAP Kinase Signaling System/genetics , Monocytes/immunology , Monocytes/metabolism , Oligonucleotide Array Sequence Analysis , RNA Interference , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolismABSTRACT
BACKGROUND AND AIMS: Orthotopic liver transplantation (OLT) can be an effective treatment option for certain patients with early stage hepatocellular carcinoma (HCC) meeting Milan, UCSF, or Hangzhou criteria. However, HCC recurrence rates post-OLT range from 20 to 40 %, with limited follow-up options. Elucidating genetic drivers common to primary and post-OLT recurrent tumors may further our understanding and help identify predictive biomarkers of recurrence-both to ultimately help manage clinical decisions for patients undergoing OLT. METHODS: Whole exome and RNA sequencing in matched primary and recurrent tumors, normal adjacent tissues, and blood from four Chinese HCC patients was conducted. SiRNA knockdown and both qRT-PCR and Western assays were performed on PLCPRF5, SNU449 and HEPG2 cell lines; immunohistochemistry and RNA Sequencing were conducted on the primary tumors of Chinese HCC patients who experienced tumor recurrence post-OLT (n = 9) or did not experience tumor recurrence (n = 12). RESULTS: In three independent HCC studies of patients undergoing transplantation (n = 21) or surgical resection (n = 242, n = 44) of primary tumors (total n = 307), HERC5 mRNA under-expression correlated with shorter: time to tumor recurrence (p = 0.007 and 0.02) and overall survival (p = 0.0063 and 0.023), even after adjustment for relevant clinical variables. HERC5 loss drives CCL20 mRNA and protein over-expression and associates with regulatory T cell infiltration as measured by FOXP3 expression. Further, matched primary and recurrent tumors from the 4 HCC patients indicated clonal selection advantage of Wnt signaling activation and CDKN2A inactivation. CONCLUSIONS: HERC5 plays a crucial role in HCC immune evasion and has clinical relevance as a reproducible prognostic marker for risk of tumor recurrence and survival in patients.
Subject(s)
Biomarkers, Tumor/metabolism , Carcinoma, Hepatocellular/surgery , Intracellular Signaling Peptides and Proteins/genetics , Liver Neoplasms/surgery , Liver Transplantation , DNA Copy Number Variations , Humans , Prognosis , RecurrenceABSTRACT
OBJECTIVE: To assess the pharmacodynamic effects of sifalimumab, an investigational anti-IFN-α monoclonal antibody, in the blood and muscle of adult dermatomyositis and polymyositis patients by measuring neutralisation of a type I IFN gene signature (IFNGS) following drug exposure. METHODS: A phase 1b randomised, double-blinded, placebo controlled, dose-escalation, multicentre clinical trial was conducted to evaluate sifalimumab in dermatomyositis or polymyositis patients. Blood and muscle biopsies were procured before and after sifalimumab administration. Selected proteins were measured in patient serum with a multiplex assay, in the muscle using immunohistochemistry, and transcripts were profiled with microarray and quantitative reverse transcriptase PCR assays. A 13-gene IFNGS was used to measure the pharmacological effect of sifalimumab. RESULTS: The IFNGS was suppressed by a median of 53-66% across three time points (days 28, 56 and 98) in blood (p=0.019) and 47% at day 98 in muscle specimens post-sifalimumab administration. Both IFN-inducible transcripts and proteins were prevalently suppressed following sifalimumab administration. Patients with 15% or greater improvement from baseline manual muscle testing scores showed greater neutralisation of the IFNGS than patients with less than 15% improvement in both blood and muscle. Pathway/functional analysis of transcripts suppressed by sifalimumab showed that leucocyte infiltration, antigen presentation and immunoglobulin categories were most suppressed by sifalimumab and highly correlated with IFNGS neutralisation in muscle. CONCLUSIONS: Sifalimumab suppressed the IFNGS in blood and muscle tissue in myositis patients, consistent with this molecule's mechanism of action with a positive correlative trend between target neutralisation and clinical improvement. These observations will require confirmation in a larger trial powered to evaluate efficacy.
Subject(s)
Antibodies, Monoclonal/administration & dosage , Dermatomyositis/drug therapy , Dermatomyositis/immunology , Immunosuppressive Agents/administration & dosage , Polymyositis/drug therapy , Polymyositis/immunology , Adult , Aged , Antibodies, Monoclonal/adverse effects , Antibodies, Monoclonal, Humanized , Double-Blind Method , Female , Gene Expression/drug effects , Gene Expression/immunology , Humans , Immunosuppressive Agents/adverse effects , Interferon Type I/blood , Interferon Type I/genetics , Interferon Type I/immunology , Interferon-alpha/blood , Interferon-alpha/genetics , Interferon-alpha/immunology , Leukocytes/drug effects , Leukocytes/immunology , Male , Middle Aged , Muscle, Skeletal/immunology , Placebos , Treatment Outcome , Young AdultABSTRACT
OBJECTIVE: The aim of this study was to identify serum markers that are modulated by an investigational anti-IFN-α mAb, sifalimumab, in adult DM or PM patients. METHODS: In a phase 1b clinical trial, sera were collected from a total of 48 DM or PM adult patients receiving either placebo for 3 months or sifalimumab for 6 months. Samples were tested for 128 selected proteins using a multiplex luminex immunoassay. Muscle biopsies from selected patients were stained for T cell infiltration using an anti-CD3 antibody. RESULTS: A robust overexpression of multiple serum proteins in DM or PM patients was observed, particularly in patients with an elevated baseline type I IFN gene signature in the blood or muscle. Neutralization of the type I IFN gene signature by sifalimumab resulted in coordinated suppression of T cell-related proteins such as soluble IL-2RA, TNF receptor 2 (TNFR2) and IL-18. Muscle biopsies from two patients with the highest serum protein suppression were selected and found to have a pronounced reduction of muscle T cell infiltration. Down-regulation of IL-2RA correlated with favourable manual muscle test 8 (MMT-8) alterations in sifalimumab-dosed patients. CONCLUSION: A reduced level of multiple T cell-associated proteins after sifalimumab but not placebo administration suggests a suppressive effect of blocking type I IFN signalling on T cell activation and chemoattraction that may lead to a reduction of T cell infiltration in the muscle of myositis patients. Further, soluble IL-2RA changes from baseline may serve as a responsive and/or predictive marker for type I IFN-targeted therapy in adult DM or PM patients.
Subject(s)
Antibodies, Monoclonal/pharmacology , Dermatomyositis/immunology , Interferon-alpha/antagonists & inhibitors , Polymyositis/immunology , T-Lymphocytes/immunology , Angiopoietin-2/immunology , Antibodies, Monoclonal/therapeutic use , Antibodies, Monoclonal, Humanized , Dermatomyositis/drug therapy , Double-Blind Method , Down-Regulation , Female , Humans , Interferon-alpha/genetics , Interleukin-18/immunology , Interleukin-2 Receptor alpha Subunit/drug effects , Interleukin-2 Receptor alpha Subunit/immunology , Male , Middle Aged , Muscle, Skeletal/drug effects , Muscle, Skeletal/immunology , Polymyositis/drug therapy , Receptors, Tumor Necrosis Factor, Type II/drug effects , Receptors, Tumor Necrosis Factor, Type II/immunology , Severity of Illness Index , T-Lymphocytes/drug effects , Treatment OutcomeABSTRACT
BACKGROUND: To evaluate the insulin receptor isoform mRNA expression status in non-small cell lung cancer (NSCLC) patients. METHODS: RNA-seq data from 614 NSCLC [355 adenocarcinomas (LUAD) and 259 squamous cell carcinomas (LUSC)] and 92 normal lung specimens were obtained from The Cancer Genome Atlas (TCGA) to evaluate the mRNA expression of insulin receptor isoform A (IR-A) and insulin receptor isoform B (IR-B). The differential expression status of the insulin receptor isoforms in NSCLC patients was confirmed using qRT-PCR assays with lung cancer cDNA arrays and primary tumor samples. RESULTS: The mRNA expression levels of IR-B were significantly lower in some NSCLC samples compared to normal lung specimens, including both LUAD and LUSC. Notably, no IR-B transcripts were detected - only the IR-A isoform was expressed in 11% of NSCLC patients. This decrease in IR-B expression contributed to an elevated IR-A/IR-B ratio, which was also associated with lower epithelial-mesenchymal transition gene signatures in NSCLC and longer patient survival under standard of care in LUSC. In addition to NSCLC, RNA-seq data from TCGA revealed a similar increase in IR-A/IR-B ratio in many other cancer types, with high prevalence in acute myeloid leukemia, glioblastoma multiforme, and brain lower grade glioma. CONCLUSIONS: Our results indicate a common reduction of the mRNA expression level of IR-B and an increased IR-A/IR-B mRNA ratio in NSCLC and other tumor types. The relationship of altered IR-A/IR-B ratios with cancer progression and patient survival should be prospectively explored in future studies.
Subject(s)
Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/pathology , Epithelial-Mesenchymal Transition/genetics , Lung Neoplasms/genetics , Lung Neoplasms/pathology , RNA Isoforms , Receptor, Insulin/genetics , Alternative Splicing , Carcinoma, Non-Small-Cell Lung/mortality , Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/mortality , Carcinoma, Squamous Cell/pathology , Computational Biology/methods , Databases, Nucleic Acid , Gene Expression Regulation, Neoplastic , Humans , Lung Neoplasms/mortality , Outcome Assessment, Health Care , Prognosis , RNA, Messenger/geneticsABSTRACT
OBJECTIVE: To evaluate the safety and tolerability of multiple intravenous (IV) doses of sifalimumab in adults with moderate-to-severe systemic lupus erythematosus (SLE). METHODS: In this multicenter, double-blind, placebo-controlled, sequential dose-escalation study, patients were randomized 3:1 to receive IV sifalimumab (0.3, 1.0, 3.0, or 10.0 mg/kg) or placebo every 2 weeks to week 26, then followed up for 24 weeks. Safety assessment included recording of treatment-emergent adverse events (AEs) and serious AEs. Pharmacokinetics, immunogenicity, and pharmacodynamics were evaluated, and disease activity was assessed. RESULTS: Of 161 patients, 121 received sifalimumab (26 received 0.3 mg/kg; 25, 1.0 mg/kg; 27, 3.0 mg/kg; and 43, 10 mg/kg) and 40 received placebo. Patients were predominantly female (95.7%). At baseline, patients had moderate-to-severe disease activity (mean SLE Disease Activity Index score 11.0), and most (75.2%) had a high type I interferon (IFN) gene signature. In the sifalimumab group versus the placebo group, the incidence of ≥1 treatment-emergent AE was 92.6% versus 95.0%, ≥1 serious AE was 22.3% versus 27.5%, and ≥1 infection was 67.8% versus 62.5%; discontinuations due to AEs occurred in 9.1% versus 7.5%, and death occurred in 3.3% (n=4) versus 2.5% (n=1). Serum sifalimumab concentrations increased in a linear and dose-proportional manner. Inhibition of the type I IFN gene signature was sustained during treatment in patients with a high baseline signature. No statistically significant differences in clinical activity (SLEDAI and British Isles Lupus Assessment Group score) between sifalimumab and placebo were observed. However, when adjusted for excess burst steroids, SLEDAI change from baseline showed a positive trend over time. A trend toward normal complement C3 or C4 level at week 26 was seen in the sifalimumab groups compared with baseline. CONCLUSION: The observed safety/tolerability and clinical activity profile of sifalimumab support its continued clinical development for SLE.
Subject(s)
Antibodies, Monoclonal/administration & dosage , Immunologic Factors/administration & dosage , Lupus Erythematosus, Systemic/drug therapy , Adult , Antibodies, Monoclonal/adverse effects , Antibodies, Monoclonal/pharmacokinetics , Antibodies, Monoclonal, Humanized , Dose-Response Relationship, Drug , Female , Humans , Immunologic Factors/adverse effects , Immunologic Factors/pharmacokinetics , Interferon-alpha/antagonists & inhibitors , Lupus Erythematosus, Systemic/metabolism , Male , Middle Aged , Severity of Illness Index , Treatment OutcomeABSTRACT
OBJECTIVE: MicroRNAs (miRNAs) function to fine-tune the control of immune cell signaling. It is well established that there are abnormalities in the interleukin-2 (IL-2)-related signaling pathways in systemic lupus erythematosus (SLE). The miR-31 microRNA has been found to be markedly underexpressed in patients with SLE, and thus the present study was undertaken to investigate the role of miR-31 in IL-2 defects in lupus T cells. METHODS: Expression levels of miR-31 were quantitated using TaqMan miRNA assays. Transfection and stimulation of cultured cells followed by TaqMan quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, and reporter gene assays were conducted to determine the biologic function of miR-31. NF-AT nuclear translocation and expression were quantitatively measured using an ImageStream cytometer. Bioinformatics analysis, small interfering RNA (siRNA) knockdown, and Western blotting were performed to validate miR-31 targets and effects. RESULTS: The expression of miR-31 was significantly decreased in lupus T cells, and this was positively correlated with the expression of IL-2. Overexpression of miR-31 in T cells increased the production of IL-2 by altering NF-AT nuclear expression and IL2 promoter activity, while knockdown of endogenous miR-31 reduced IL-2 production. RhoA expression was directly repressed by miR-31 in T cells. Of note, siRNA-mediated knockdown of RhoA enhanced IL2 promoter activity and, consequently, up-regulated IL-2 production. RhoA expression was consistently up-regulated and negatively correlated with the levels of miR-31 in lupus T cells. Manipulation of miR-31 expression in lupus T cells restored the expression of IL-2 at both the messenger RNA and protein levels. CONCLUSION: MicroRNA-31 is a novel enhancer of IL-2 production during T cell activation. Dysregulation of miR-31 and its target, RhoA, could be a novel molecular mechanism underlying the IL-2 deficiency in patients with SLE.
Subject(s)
Interleukin-2/genetics , Lupus Erythematosus, Systemic/genetics , Lupus Erythematosus, Systemic/immunology , MicroRNAs/immunology , T-Lymphocytes/immunology , Gene Expression Regulation/genetics , Gene Expression Regulation/immunology , Humans , Interleukin-2/deficiency , Interleukin-2/immunology , Jurkat Cells , Lupus Erythematosus, Systemic/metabolism , NFATC Transcription Factors/genetics , NFATC Transcription Factors/immunology , Primary Cell Culture , Promoter Regions, Genetic/genetics , Promoter Regions, Genetic/immunology , RNA, Small Interfering/genetics , Signal Transduction/genetics , Signal Transduction/immunology , T-Lymphocytes/metabolism , rhoA GTP-Binding Protein/genetics , rhoA GTP-Binding Protein/immunologyABSTRACT
BACKGROUND: Anti-B-cell maturation antigen (BCMA) chimeric antigen receptor T-cell (CAR T) therapy showed remarkable efficacy in patients with relapsed or refractory multiple myeloma (RRMM). This phase 1 dose-escalation and expansion study developed C-CAR088, a novel second-generation humanized anti-BCMA CAR T-cell therapy, and assessed the safety and efficacy of three dosages of C-CAR088 in patients with RRMM. METHODS: Patients received lymphodepletion with three doses of cyclophosphamide (300 mg/m2) and three doses of fludarabine (30 mg/m2) on days -5, -4, and -3, followed by an infusion of C-CAR088 on day 0. Doses of 1.0×106, 3.0×106, and 6.0×106 CAR T cells/kg (±20%) were tested in the dose-escalation cohorts and expansion cohorts. The primary endpoint was treatment safety, including the rate of treatment-emergent adverse events after cell infusion. Secondary endpoints were the overall response rate and progression-free survival. The exploratory endpoints were the quantification of C-CAR088 CAR T cells, selection of cytokines and chemokines in blood, and measurement of tumor BCMA expression. RESULTS: As of July 2, 2021, 31 patients had been infused with C-CAR088. Any grade cytokine release syndrome (CRS) occurred in 29 patients (93.5%), and grade 3 CRS occurred in 3 patients (9.7%). One patient from the high-dose group (4.5-6.0×106 CAR T cells/kg) developed grade 1 neurotoxicity. No dose-limiting toxicities were observed in any dose group, and all adverse events were reversible after proper management. The overall response, stringent complete response, complete response (CR), and very good partial response rates were 96.4%, 46.4%, 10.7%, and 32.1%, respectively. The CR rate in the medium-dose (3.0×106 CAR T cells/kg) and high-dose (4.5-6.0×106 CAR T cells/kg) groups was 54.5% and 71.4%, respectively. In the CR group, 15 (93.7%) patients achieved minimal residual disease (MRD) negativity (test sensitivity >1/10-5). All seven patients with double-hit or triple-hit multiple myeloma achieved MRD-negative CR. CONCLUSIONS: The present study demonstrated that C-CAR088 had a good safety profile and high antitumor activity in patients with RRMM, constituting a promising treatment option for RRMM. TRIAL REGISTRATION NUMBER: NCT03815383, NCT03751293, NCT04295018, and NCT04322292.
Subject(s)
Multiple Myeloma , Receptors, Chimeric Antigen , Cyclophosphamide , Humans , Immunotherapy, Adoptive/adverse effects , T-LymphocytesABSTRACT
BACKGROUND: Type I interferons (IFNs) appear to play a central role in disease pathogenesis in systemic lupus erythematosus (SLE), making them potential therapeutic targets. METHODS: Safety profile, pharmacokinetics, immunogenicity, pharmacodynamics and clinical activity of sifalimumab, an anti-IFNα monoclonal antibody, were assessed in a phase I, multicentre, randomised, double-blind, dose-escalation study with an open-label extension in adults with moderately active SLE. SUBJECTS: received one intravenous dose of sifalimumab (n=33 blinded phase, 0.3, 1, 3, 10 or 30 mg/kg; n=17 open-label, 1, 3, 10 or 30 mg/kg) or placebo (n=17). Each phase lasted 84 days. RESULTS: Adverse events (AEs) were similar between groups; about 97% of AEs were grade 1 or 2. All grade 3 and 4 AEs and all serious AEs (2 placebo, 1 sifalimumab) were deemed unrelated to the study drug. No increase in viral infections or reactivation was observed. Sifalimumab caused dose-dependent inhibition of type I IFN-induced mRNAs (type I IFN signature) in whole blood and corresponding changes in related proteins in affected skin. Exploratory analyses showed consistent trends toward improvement in disease activity in sifalimumab-treated versus placebo-treated subjects. A lower proportion of sifalimumab-treated subjects required new or increased immunosuppressive treatments (12% vs 41%; p=0.03) and had fewer Systemic Lupus Erythematosus Disease Activity Index flares (3% vs 29%; p=0.014). CONCLUSIONS: Sifalimumab had a safety profile that supports further clinical development. This trial demonstrated that overexpression of type I IFN signature in SLE is at least partly driven by IFNα, and exploratory analyses suggest that IFNα inhibition may be associated with clinical benefit in SLE. Trial registration number NCT00299819.
Subject(s)
Antibodies, Monoclonal/adverse effects , Immunosuppressive Agents/adverse effects , Interferon-alpha/antagonists & inhibitors , Lupus Erythematosus, Systemic/drug therapy , Adult , Aged , Aged, 80 and over , Antibodies, Monoclonal/administration & dosage , Antibodies, Monoclonal/blood , Antibodies, Monoclonal/therapeutic use , Antibodies, Monoclonal, Humanized , Dose-Response Relationship, Drug , Double-Blind Method , Female , Gene Expression Regulation/drug effects , Humans , Immunosuppressive Agents/administration & dosage , Immunosuppressive Agents/blood , Immunosuppressive Agents/therapeutic use , Injections, Intravenous , Interferon Type I/biosynthesis , Interferon Type I/genetics , Interferon-alpha/immunology , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/immunology , Male , Middle Aged , RNA, Messenger/genetics , Severity of Illness Index , Treatment Outcome , Young AdultABSTRACT
OBJECTIVE: To characterise activation of the type I interferon (IFN) pathway in patients with systemic lupus erythematosus (SLE), dermatomyositis (DM), polymyositis (PM), rheumatoid arthritis (RA) and systemic scleroderma (SSc) and to evaluate the potential to develop a molecular diagnostic tool from the peripheral blood that reflects this activation in disease-affected tissues. METHODS: Overexpressed transcripts were identified in the whole blood (WB) of 262 patients with SLE, 44 with DM, 33 with PM, 28 with SSc and 89 with RA and compared with 24 healthy subjects using Affymetrix microarrays. A five gene type I IFN signature was assessed in these subjects to identify subpopulations showing both activation and concordance of the type I IFN pathway in the peripheral blood and disease-affected tissues of each disease and to correlate activation of this pathway in the WB with clinical measurements. RESULTS: A common set of 36 type I IFN inducible transcripts were identified among the most overexpressed in the WB of all subjects. Significant activation of the type I IFN pathway in subgroups of each of the five diseases studied was observed. Baseline disease activity measurements correlated with a type I IFN gene signature in the WB of subjects with SLE, PM and SSc, as did various serum autoantibody levels in subjects with SLE and DM. This signature was also well correlated between disease-affected tissue and WB in subjects with SLE, DM, PM and SSc. CONCLUSIONS: The results indicate that the type I IFN pathway is activated in patient subsets of five rheumatic diseases and suggest that these subsets may benefit from anti-IFN therapy.
Subject(s)
Interferon Type I/biosynthesis , Rheumatic Diseases/immunology , Adult , Arthritis, Rheumatoid/immunology , Biomarkers/blood , Blood Proteins/metabolism , Female , Gene Expression Profiling/methods , Humans , Interferon Type I/genetics , Interferon-alpha/antagonists & inhibitors , Lupus Erythematosus, Systemic/immunology , Male , Middle Aged , Myositis/immunology , Oligonucleotide Array Sequence Analysis/methods , Scleroderma, Systemic/immunology , Severity of Illness Index , Signal Transduction/immunologyABSTRACT
OBJECTIVE: We investigated interferon-stimulated gene 15 (ISG15), a poorly understood ubiquitin-like modifier, and its enzymatic pathway in dermatomyositis (DM), an autoimmune disease primarily involving muscle and skin. METHODS: We generated microarray data measuring transcript abundance for approximately 18,000 genes in each of 113 human muscle biopsy specimens, and studied biopsy specimens and cultured skeletal muscle using immunohistochemistry, immunoblotting proteomics, real-time quantitative polymerase chain reaction, and laser-capture microdissection. RESULTS: Transcripts encoding ISG15-conjugation pathway proteins were markedly upregulated in DM with perifascicular atrophy (DM-PFA) muscle (ISG15 339-fold, HERC5 62-fold, and USP18 68-fold) compared with 99 non-DM samples. Combined analysis with publicly available microarray datasets showed that >50-fold ISG15 transcript elevation had 100% sensitivity and specificity for 28 biopsies from adult DM-PFA and juvenile DM patients compared with 199 muscle samples from other muscle diseases. Free ISG15 and ISG15-conjugated proteins were only found on immunoblots from DM-PFA muscle. Cultured human skeletal muscle exposed to type 1 interferons produced similar transcripts and ISG15 protein and conjugates. Laser-capture microdissection followed by proteomic analysis showed deficiency of titin in DM perifascicular atrophic myofibers. INTERPRETATION: A large-scale microarray study of muscle samples demonstrated that among a diverse group of muscle diseases DM was uniquely associated with upregulation of the ISG15 conjugation pathway. Exposure of human skeletal muscle cell culture to type 1 interferons produced a molecular picture highly similar to that seen in human DM muscle. Perifascicular atrophic myofibers in DM were deficient in a number of skeletal muscle proteins including titin.
Subject(s)
Cytokines/metabolism , Dermatomyositis/metabolism , Muscle, Skeletal/metabolism , Ubiquitins/metabolism , Cells, Cultured , Connectin , Cytokines/genetics , Databases, Genetic , Dermatomyositis/diagnosis , Dermatomyositis/genetics , Humans , Immunoblotting , Immunohistochemistry , Interferon Type I/metabolism , Lasers , Microdissection/methods , Muscle Proteins/deficiency , Muscular Diseases/diagnosis , Muscular Diseases/genetics , Muscular Diseases/metabolism , Oligonucleotide Array Sequence Analysis , Polymerase Chain Reaction , Protein Kinases/deficiency , Proteomics/methods , Sensitivity and Specificity , Ubiquitins/genetics , Up-RegulationABSTRACT
To determine the potential consequences of plasmacytoid dendritic cell (pDC) accumulation in tissue sites observed in several autoimmune diseases, we measured type 1 interferon production from circulating human pDCs as a function of pDC concentration. The effects of interferon-alpha and blockade of the type 1 interferon receptor (IFNAR) on human pDC type 1 interferon and interferon-inducible transcription and protein production were measured. Human pDCs became far more efficient producers of interferon-alpha at concentrations beyond those normally present in blood, through an IFNAR-dependent mechanism. Extracellular interferon-alpha increased pDC production of type 1 interferons. The accumulation of pDCs in diseased tissue sites allows marked non-linear amplification of type 1 interferon production locally. The role of the IFNAR-dependent mechanism of interferon production by human pDCs is greater than previously suggested. IFNAR blockade has potential for diminishing type 1 interferon production by all human cells.
Subject(s)
Dendritic Cells/cytology , Dendritic Cells/metabolism , Interferon Type I/metabolism , Adaptor Proteins, Signal Transducing , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/pharmacology , Autocrine Communication/drug effects , Autocrine Communication/immunology , Carrier Proteins/genetics , Cell Count , Cytokines/genetics , Cytokines/metabolism , Dendritic Cells/drug effects , Endopeptidases/genetics , GTP-Binding Proteins/genetics , GTP-Binding Proteins/metabolism , Gene Expression/drug effects , Gene Expression/genetics , Humans , Interferon Type I/genetics , Interferon alpha-2 , Interferon-alpha/genetics , Interferon-alpha/metabolism , Interferon-alpha/pharmacology , Interferon-beta/genetics , Intracellular Signaling Peptides and Proteins/genetics , Kinetics , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/metabolism , Myxovirus Resistance Proteins , Oligodeoxyribonucleotides/pharmacology , Oxidoreductases Acting on CH-CH Group Donors , Protein Isoforms/genetics , Proteins/genetics , RNA-Binding Proteins , Receptor, Interferon alpha-beta/antagonists & inhibitors , Receptor, Interferon alpha-beta/immunology , Recombinant Proteins , Toll-Like Receptor 9/agonists , Transcription, Genetic/drug effects , Transcription, Genetic/genetics , Ubiquitin Thiolesterase , Ubiquitins/genetics , Ubiquitins/metabolismABSTRACT
Myositis muscle contains antigen-matured B-cells and plasma cells. Myositis muscle biopsy specimens were examined for nodular collections of T-cells, B-cells, myeloid dendritic cells, plasma cells, and follicular dendritic cells. Immunoglobulin and B-cell-activating factor (BAFF) transcripts were quantitated. Laser-capture microdissection was used to isolate single plasma cells, and their immunoglobulin transcripts were sequenced. Dense inflammatory infiltrates contained histological elements of ectopic lymphoid tissue but not B-cell follicles. Immunoglobulin transcript sequence analysis demonstrated spatially distributed, clonally related B-cells and plasma cells, suggesting local maturation of B-cells into plasma cells in myositis muscle. Regions of dense cellular infiltrates in myositis muscle are sometimes areas of B-cell maturation into antibody-producing plasma cells. An atypical lymphoid histology, lacking concentrated collections of germinal-center-like B-cell follicles, is capable of antigen-stimulated clonal maturation of antibody-producing plasma cells.