The effect of human umbilical cord mesenchymal stem cell-derived exosomes on diabetic retinal neurodegeneration in a rat model.

OBJECTIVE: To investigate the effect of human umbilical cord mesenchymal stem cell-derived exosomes (hucMSC-Exs) on diabetic retinal neurodegeneration (DRN). METHODS: Exosomes were isolated from human umbilical cord mesenchymal stem cells (hucMSC) and identified using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and Western blotting (WB). Rats were intraperitoneally injected with Streptozotocin (STZ) to establish a diabetes mellitus model, and blood glucose levels and body weight were assessed. The rats were intravitreally injected with phosphate buffered saline (PBS; diabetic group) or hucMSC-Exs (hucMSC-Exs group). A control group of rats were not treated with STZ and were intravitreally injected with PBS (normal control group). Hematoxylin-eosin (HE) staining was used to observe changes in retinal structure and to count the number of retinal ganglion cells (RGCs) four weeks after intravitreal injection. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay (TUNEL) was used to detect retinal cell apoptosis. The retinal expression of p38 mitogen-activated protein kinase (p38MAPK), phosphorylated p38MAPK (p-p38MAPK), Bcl-2 and Bax was measured using WB to investigate the mechanism by which hucMSC-Exs affects DRN. RESULTS: Using TEM, NTA and WB, hucMSC-Exs were successfully isolated. No significant change was observed after injection in the normal control group. All rats injected with STZ developed hyperglycemia. HE staining revealed that hucMSC-Exs effectively alleviated retinal structure disruption and reduced the apoptosis of RGCs (P < 0.05). Cells positive for TUNEL (TUNEL+) occurred at a higher rate in the diabetic group than in other groups (P < 0.05). Compared with the normal control group, the expression of p-p38MAPK was significantly increased in the diabetic group and decreased in the hucMSC-Exs group (P < 0.01). The expression of Bax was significantly decreased while Bcl-2 expression was significantly increased in hucMSC-Exs group (P < 0.01). CONCLUSION: These findings suggest that intravitreal injection of hucMSC-Exs can reduce DRN and protect retinal structure, and that these effects are mediated through inhibition of the p38MAPK pathway.

The development of epiretinal membrane following rhegmatogenous retinal detachment repair: incidence, risk factors, and outcomes / Desenvolvimento da membrana epirretiniana após reparo do descolamento regmatogênico da retina: incidência, fatores de risco e resultados

ABSTRACT Purpose: To investigate the incidence, risk factors, and visual outcomes of epiretinal membrane development following rhegmatogenous retinal detachment repair. Methods: This was a retrospective study of 309 eyes that underwent initial surgery for primary uncomplicated rhegmatogenous retinal detachment. Examinations were conducted preoperatively and then postoperatively at 1, 3, 6, and 12 months. The study patients were categorized into two groups depending on the presence or absence of the epiretinal membrane. Results: The incidence of postoperative epiretinal membrane was 28.5%; 42.7% of these patients had severe epiretinal membrane development and therefore underwent the epiretinal membrane removal. Logistic regression analyses revealed that giant retinal tears (OR: 2.66; 95% CI: 1.045-6.792, p=0.040) and horseshoe tears (OR: 0.534; 95% CI: 0.295-0.967, p=0.039) were the significant predictors of postoperative epiretinal membrane. Triamcinolone acetonide staining was significantly associated with the prevention of epiretinal membrane (p=0.022). A total of 34 patients showed a better or an equal final best-corrected visual acuity; of which 4 eyes were evaluated at the final follow-up visit and exhibited a reduced best-corrected visual acuity. Conclusion: Our analysis demonstrated that horseshoe tears and giant retinal tears represent the risk factors for the postoperative epiretinal membrane. Triamcinolone acetonide staining had a significant preventive effect on the postoperative epiretinal membrane. Furthermore, a second round of pars plana vitrectomy, including membrane removal, led to a significant improvement in the final best-corrected visual acuity as per the last follow-up examination, albeit the recovery was limited.

RESUMO Objetivos: Investigar a incidência, fatores de risco e desfechos visuais do desenvolvimento da membrana epirretiniana após reparo do descolamento regmatogênico da retina. Métodos: Trata-se de um estudo retrospectivo de 309 olhos submetidos à cirurgia inicial para descolamento regmatogênico da retina primário sem complicações. Os exames foram realizados no pré-operatório aos 1, 3, 6 e 12 meses pós-operatórios. Os pacientes foram divididos em dois grupos, dependendo da presença ou ausência de membrana epirretiniana. Resultados: A incidência de membrana epirretiniana pós-operatória foi de 28,5%; 42,7% desses pacientes apresentaram desenvolvimento grave da membrana epirretiniana e, portanto, foram submetidos à remoção desta membrana. A regressão logística mostrou que as lágrimas retinianas gigantes (RC: 2,66; 95% IC: 1,045 - 6,792, p=0,040) e lágrimas em ferradura (RC: 0,534; 95% IC: 0,295-0,967, p=0,039), foram preditores significativos de membrana epirretiniana pós-operatória. A coloração com acetonida de triancinolona foi significativamente associada à prevenção da membrana epirretiniana (p=0,022). Trinta e quatro pacientes apresentaram acuidade visual melhorada, ou igual, ou acuidade visual final melhor corrigida; 4 olhos foram avaliados na consulta final de acompanhamento e apresentaram redução da acuidade visual melhor corrigida. Conclusão: Nossa análise demonstra que as lágrimas de ferradura e as lágrimas retinianas gigantes representam fatores de risco para a membrana epirretiniana pós-operatória. A coloração com acetonida de triancinolona teve um efeito preventivo significativo na membrana epirretiniana no pós-operatório. Além disso, uma segunda rodada de vitrectomia pars plana, incluindo remoção da membrana, levou a uma melhora significativa da acuidade visual final melhor corrigida na última consulta de acompanhamento, embora a recuperação tenha sido limitada.

Recurrent retinal detachment after pars plana vitrectomy with silicone oil tamponade for rhegmatogenous retinal detachment.

BACKGROUND: The recurrence of retinal detachment following rhegmatogenous retinal detachment (RRD) is a relatively common complication that can lead to reduced visual acuity and requires further surgery. The purpose of this study was to investigate the risk factors and visual outcomes of recurrent RRD following pars plana vitrectomy (PPV) with silicone oil tamponade for primary RRD. METHODS: This was a retrospective follow-up study of 343 eyes that underwent initial PPV surgery with silicone oil tamponade for primary RRD. Patients were divided into a recurrence group and a reattachment group. The main outcome measures included causative factors, visual outcomes related to the recurrence of RRD, and the perioperative factors most affecting the recurrence of RRD. RESULTS: After retinal reattachment, we observed RRD recurrence after PPV for primary RRD in 42 out of 343 eyes (12.2%) during the follow-up period. Most causes of recurrence (69%) occurred within 6 months of surgery. Multivariate logistic regression analysis showed that a PVR ≥ Grade C (odds ratio [OR]: 4.015; 95% confidence interval [CI] 1.721-9.367; P = 0.001) was a significant predictor for the development of recurrent RRD. Compared with the reattachment group, the recurrence group exhibited a significant decline in best-corrected visual acuity (BCVA) at the last follow-up visit (P = 0.000). Eyes with PVR prior to primary surgery, or at the diagnosis of re-detachment, showed a worse final BCVA. CONCLUSIONS: Our analysis shows that the predominant risk factor for the recurrence of RRD is a PVR ≥ Grade C. PVR prior to primary surgery, or at the diagnosis of re-detachment, was also shown to limit the recovery of final visual acuity.

Protective effects of umbilical cord mesenchymal stem cell exosomes in a diabetic rat model through live retinal imaging.

AIM: To assess the protective effect of human umbilical cord mesenchymal stem cell exosomes (hucMSC-Exs) in a diabetic rat model by using a variety of retinal bioassays. METHODS: hucMSCs were subjected to differential ultracentrifugation for the collection of exosomes, and transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) using a NanoSight analysis system and Western blotting (WB) were used to analyze the expression of surface marker proteins such as CD63, CD9 and Calnexin. Streptozotocin (STZ) was injected into the intraperitoneal cavity to establish a diabetic model. Rats were divided into a normal group, diabetic group and hucMSC-Ex group. Fundus fluorescein angiography (FFA), optical coherence tomography (OCT) and other live imaging methods were used to observe the fundus of the rats. Finally, the eyeballs of rats from each group were collected for hematoxylin-eosin (HE) staining to further analyze the retinal structure. RESULTS: Through TEM, NTA and WB, we successfully isolated hucMSC-Exs. Subsequent FFA and OCT confirmed that hucMSC-Exs effectively prevented early retinal vascular damage and thickening of the retina. Finally, HE staining of rat retinal sections revealed that exosomes effectively alleviated retinal structure disruption caused by diabetes. CONCLUSION: hucMSC-Exs have a protective effect on the retina in diabetic rat through FFA, OCT and HE staining.

The development of epiretinal membrane following rhegmatogenous retinal detachment repair: incidence, risk factors, and outcomes.

PURPOSE: To investigate the incidence, risk factors, and visual outcomes of epiretinal membrane development following rhegmatogenous retinal detachment repair. METHODS: This was a retrospective study of 309 eyes that underwent initial surgery for primary uncomplicated rhegmatogenous retinal detachment. Examinations were conducted preoperatively and then postoperatively at 1, 3, 6, and 12 months. The study patients were categorized into two groups depending on the presence or absence of the epiretinal membrane. RESULTS: The incidence of postoperative epiretinal membrane was 28.5%; 42.7% of these patients had severe epiretinal membrane development and therefore underwent the epiretinal membrane removal. Logistic regression analyses revealed that giant retinal tears (OR: 2.66; 95% CI: 1.045-6.792, p=0.040) and horseshoe tears (OR: 0.534; 95% CI: 0.295-0.967, p=0.039) were the significant predictors of postoperative epiretinal membrane. Triamcinolone acetonide staining was significantly associated with the prevention of epiretinal membrane (p=0.022). A total of 34 patients showed a better or an equal final best-corrected visual acuity; of which 4 eyes were evaluated at the final follow-up visit and exhibited a reduced best-corrected visual acuity. CONCLUSION: Our analysis demonstrated that horseshoe tears and giant retinal tears represent the risk factors for the postoperative epiretinal membrane. Triamcinolone acetonide staining had a significant preventive effect on the postoperative epiretinal membrane. Furthermore, a second round of pars plana vitrectomy, including membrane removal, led to a significant improvement in the final best-corrected visual acuity as per the last follow-up examination, albeit the recovery was limited.

Natural history of persistent subretinal fluid following the successful repair of rhegmatogenous retinal detachment.

AIM: To provide a detailed description of the natural history of persistent subretinal fluid (SRF) after successful repair of rhegmatogenous retinal detachment (RRD) and its association with visual outcome. METHODS: This was a prospective long-term follow-up for eyes undergoing scleral buckling (SB) surgery for macula-off RRD. Examinations were carried out preoperatively and postoperatively at 1, 3, 6, 9 and 12mo, until persistent SRF had completely resolved. One month postoperatively, optical coherence tomography (OCT) was used to classify SRF into three patterns: bleb-like loculated (BL), shallow-diffused (SD), and multiple blebs (MB). Serial OCT imaging was used to evaluate morphological changes in SRF until its complete disappearance. Patients were divided into two groups depending on the presence or absence of persistent SRF. RESULTS: A total of 59 patients (59 eyes) were included. There were no statistical differences between two groups at baseline, except for the proportion of patients with high myopia and a younger age. One month after surgery, OCT detected persistent SRF in 49 eyes (83.1%). The 3 morphological patterns of SRF were observed in 27 eyes (55.1%) with BL, 13 eyes (26.5%) with SD, and 9 eyes (18.4%) with MB. The mean time for complete absorption differed significantly across the three SRF patterns (F=8.097, P=0.001), which was 8.8±6.1, 20.1±12.1, and 16.7±10.2mo in BL, SD, and MB, respectively. In 9 of the 13 eyes with SD, the pattern transformed into MB type. In cases involving MB, the size and number of blebs decreased gradually until they had been completely absorbed. Eyes with persistent SRF were more likely to demonstrate disruption of the ellipsoid zone (49.0% vs 10%, P=0.034). The final best-corrected visual acuity of two groups was 0.37±0.11 (with SRF) vs 0.34±0.12 (without SRF) logMAR (P=0.499), respectively. CONCLUSION: High preoperative myopia and younger age are associated with persistent SRF. BL is the most commonly observed pattern with the shortest duration and gradually disappeared. Most cases involving SD SRF transform into MB type during resolution. The size and number of the MBs decrease gradually until they were completely absorbed. The absence of persistent SRF may contribute to slow visual recovery in the short-term but does not influence the final visual outcome.

Multiregion sequencing reveals the intratumor heterogeneity of driver mutations in TP53-driven non-small cell lung cancer.

Intratumor heterogeneity (ITH) in non-small cell lung cancer (NSCLC) may account for resistance after a period of targeted therapies because drugs destroy only a portion of tumor cells. The recognition of ITH helps identify high-risk patients to make effective treatment decisions. However, ITH studies are confounded by interpatient heterogeneity in NSCLC and a large amount of passenger mutations. To address these issues, we recruited NSCLC patients carrying TP53 mutations and selected driver mutations within recurrently mutated genes in NSCLC. A total of 12-paired normal-tumor tissues were subjected to whole-genome/whole-exome sequencing. From these, 367 non-silent mutations were selected as driver mutations and deeply sequenced in 61 intratumoral microdissections. We identified a universal prevalence of heterogeneity in all 12 tumors, indicating branched evolution. Although TP53 mutations were observed in single biopsy of all 12 tumors, most tumors consist of both TP53 mutated and non-mutated cells in separate regions within the same tumor. This suggests the late molecular timing of the acquisition of TP53 mutations; therefore, the detection of TP53 mutations in a single biopsy may simply not reflect the early malignant potential. In addition, we identified regions of loss of heterozygosity surrounding TP53 and CDKN2A mutations in tumor 711, which also exhibited heterogeneity in different regional samples. Because the ITH of driver mutations likely has clinical consequences, further efforts are needed to limit the impact of ITH and to improve therapeutic efficiency, which will benefit NSCLC patients receiving targeted treatments.

Characterization of Somatic Mutations in Air Pollution-Related Lung Cancer.

Air pollution has been classified as Group 1 carcinogenic to humans, but the underlying tumorigenesis remains unclear. In Xuanwei City of Yunnan Province, the lung cancer incidence is among the highest in China attributed to severe air pollution generated by combustion of smoky coal, providing a unique opportunity to dissect lung carcinogenesis of air pollution. Here we analyzed the somatic mutations of 164 non-small cell lung cancers (NSCLCs) from Xuanwei and control regions (CR) where smoky coal was not used. Whole genome sequencing revealed a mean of 289 somatic exonic mutations per tumor and the frequent C:G â†’ A:T nucleotide substitutions in Xuanwei NSCLCs. Exome sequencing of 2010 genes showed that Xuanwei and CR NSCLCs had a mean of 68 and 22 mutated genes per tumor, respectively (p < 0.0001). We found 167 genes (including TP53, RYR2, KRAS, CACNA1E) which had significantly higher mutation frequencies in Xuanwei than CR patients, and mutations in most genes in Xuanwei NSCLCs differed from those in CR cases. The mutation rates of 70 genes (e.g., RYR2, MYH3, GPR144, CACNA1E) were associated with patients' lifetime benzo(a)pyrene exposure. This study uncovers the mutation spectrum of air pollution-related lung cancers, and provides evidence for pollution exposure-genomic mutation relationship at a large scale.

Mutations of Epigenetic Modifier Genes as a Poor Prognostic Factor in Acute Promyelocytic Leukemia Under Treatment With All-Trans Retinoic Acid and Arsenic Trioxide.

BACKGROUND: Acute promyelocytic leukemia (APL) is a model for synergistic target cancer therapy using all-trans retinoic acid (ATRA) and arsenic trioxide (ATO), which yields a very high 5-year overall survival (OS) rate of 85 to 90%. Nevertheless, about 15% of APL patients still get early death or relapse. We performed this study to address the possible impact of additional gene mutations on the outcome of APL. METHODS: We included a consecutive series of 266 cases as training group, and then validated the results in a testing group of 269 patients to investigate the potential prognostic gene mutations, including FLT3-ITD or -TKD, N-RAS, C-KIT, NPM1, CEPBA, WT1, ASXL1, DNMT3A, MLL (fusions and PTD), IDH1, IDH2 and TET2. RESULTS: More high-risk patients (50.4%) carried additional mutations, as compared with intermediate- and low-risk ones. The mutations of epigenetic modifier genes were associated with poor prognosis in terms of disease-free survival in both training (HR = 6.761, 95% CI 2.179-20.984; P = 0.001) and validation (HR = 4.026, 95% CI 1.089-14.878; P = 0.037) groups. Sanz risk stratification was associated with CR induction and OS. CONCLUSION: In an era of ATRA/ATO treatment, both molecular markers and clinical parameter based stratification systems should be used as prognostic factors for APL.

Exome sequencing identifies somatic mutations of DDX3X in natural killer/T-cell lymphoma.

Natural killer/T-cell lymphoma (NKTCL) is a malignant proliferation of CD56(+) and cytoCD3(+) lymphocytes with aggressive clinical course, which is prevalent in Asian and South American populations. The molecular pathogenesis of NKTCL has largely remained elusive. We identified somatic gene mutations in 25 people with NKTCL by whole-exome sequencing and confirmed them in an extended validation group of 80 people by targeted sequencing. Recurrent mutations were most frequently located in the RNA helicase gene DDX3X (21/105 subjects, 20.0%), tumor suppressors (TP53 and MGA), JAK-STAT-pathway molecules (STAT3 and STAT5B) and epigenetic modifiers (MLL2, ARID1A, EP300 and ASXL3). As compared to wild-type protein, DDX3X mutants exhibited decreased RNA-unwinding activity, loss of suppressive effects on cell-cycle progression in NK cells and transcriptional activation of NF-κB and MAPK pathways. Clinically, patients with DDX3X mutations presented a poor prognosis. Our work thus contributes to the understanding of the disease mechanism of NKTCL.

Genome-wide association study identifies a novel susceptibility gene for serum TSH levels in Chinese populations.

Thyroid-stimulating hormone (TSH) is a sensitive indicator of thyroid function. High and low TSH levels reflect hypothyroidism and hyperthyroidism, respectively. Even within the normal range, small differences in TSH levels, on the order of 0.5-1.0 mU/l, are associated with significant differences in blood pressure, BMI, dyslipidemia, risk of atrial fibrillation and atherosclerosis. Most of the variance in TSH levels is thought to be genetically influenced. We conducted a genome-wide association study of TSH levels in 1346 Chinese Han individuals. In the replication study, we genotyped four candidate SNPs with the top association signals in an independent isolated Chinese She cohort (n = 3235). We identified a novel serum TSH susceptibility locus within XKR4 at 8q12.1 (rs2622590, Pcombined = 2.21 × 10(-10)), and we confirmed two previously reported TSH susceptibility loci near FOXE1 at 9q22.33 and near CAPZB at 1p36.13, respectively. The rs2622590_T allele at XKR4 and the rs925489_C allele near FOXE1 were correlated with low TSH levels and were found to be nominally associated to patients with papillary thyroid carcinoma (PTC) (OR = 1.41, P= 0.014 for rs2622590_T, and OR = 1.61, P= 0.030 for rs925489_C). The rs2622590 and rs925489 genotypes were also correlated with the expression levels of FOXE1 and XKR4, respectively, in PTC tissues (P = 2.41 × 10(-4) and P= 0.02). Our findings suggest that the SNPs in XKR4 and near FOXE1 are involved in the regulation of TSH levels.

Genomic landscape of CD34+ hematopoietic cells in myelodysplastic syndrome and gene mutation profiles as prognostic markers.

Myelodysplastic syndrome (MDS) includes a group of diseases characterized by dysplasia of bone marrow myeloid lineages with ineffective hematopoiesis and frequent evolution to acute myeloid leukemia (AML). Whole-genome sequencing was performed in CD34(+) hematopoietic stem/progenitor cells (HSPCs) from eight cases of refractory anemia with excess blasts (RAEB), the high-risk subtype of MDS. The nucleotide substitution patterns were found similar to those reported in AML, and mutations of 96 protein-coding genes were identified. Clonal architecture analysis revealed the presence of subclones in six of eight cases, whereas mutation detection of CD34(+) versus CD34(-) cells revealed heterogeneity of HSPC expansion status. With 39 marker genes belonging to eight functional categories, mutations were analyzed in 196 MDS cases including mostly RAEB (n = 89) and refractory cytopenia with multilineage dysplasia (RCMD) (n = 95). At least one gene mutation was detected in 91.0% of RAEB, contrary to that in RCMD (55.8%), suggesting a higher mutational burden in the former group. Gene abnormality patterns differed between MDS and AML, with mutations of activated signaling molecules and NPM1 being rare, whereas those of spliceosome more common, in MDS. Finally, gene mutation profiles also bore prognostic value in terms of overall survival and progression free survival.

GSTT1 deletion is related to polycyclic aromatic hydrocarbons-induced DNA damage and lymphoma progression.

The interrelationship between genetic susceptibility and carcinogenic exposure is important in cancer development. Polymorphisms in detoxification enzymes of the glutathione-S-transferases (GST) family are associated with an increased incidence of lymphoma. Here we investigated the molecular connection of the genetic polymorphism of GSTT1 to the response of lymphocytes to polycyclic aromatic hydrocarbons (PAH). In neoplastic situation, GSTT1 deletions were more frequently observed in lymphoma patients (54.9%) than in normal controls (42.0%, P = 0.009), resulting in an increased risk for lymphoma in individuals with GSTT1-null genotype (Odds ratio = 1.698, 95% confidence interval = 1.145-2.518). GSTT1 gene and protein expression were accordingly decreased in GSTT1-deleting patients, consistent with activated profile of cell cycle regulation genes. Mimicking environmental exposure using long-term repeat culture with low-dose PAH metabolite Hydroquinone, malignant B- and T-lymphocytes presented increased DNA damage, pCHK1/MYC expression and cell proliferation, which were counteracted by ectopic expression of GSTT1. Moreover, GSTT1 expression retarded xenograft tumor formation of Hydroquinone-treated lymphoma cells in nude mice. In non-neoplastic situation, when zebrafish was exposed to PAH Benzo(a)pyrene, molecular silencing of gstt1 enhanced the proliferation of normal lymphocytes and upregulated myca expression. Collectively, these findings suggested that GSTT1 deletion is related to genetic predisposition to lymphoma, particularly interacting with environmental pollutants containing PAH.

DNMT3A Arg882 mutation drives chronic myelomonocytic leukemia through disturbing gene expression/DNA methylation in hematopoietic cells.

The gene encoding DNA methyltransferase 3A (DNMT3A) is mutated in ∼20% of acute myeloid leukemia cases, with Arg882 (R882) as the hotspot. Here, we addressed the transformation ability of the DNMT3A-Arg882His (R882H) mutant by using a retroviral transduction and bone marrow transplantation (BMT) approach and found that the mutant gene can induce aberrant proliferation of hematopoietic stem/progenitor cells. At 12 mo post-BMT, all mice developed chronic myelomonocytic leukemia with thrombocytosis. RNA microarray analysis revealed abnormal expressions of some hematopoiesis-related genes, and the DNA methylation assay identified corresponding changes in methylation patterns in gene body regions. Moreover, DNMT3A-R882H increased the CDK1 protein level and enhanced cell-cycle activity, thereby contributing to leukemogenesis.

Identification of BACH2 as a susceptibility gene for Graves' disease in the Chinese Han population based on a three-stage genome-wide association study.

The BACH2 gene regulates B cell differentiation and function and has been reported to be a shared susceptibility gene for several autoimmune diseases. Our previous genome-wide association study (GWAS) indicated that several single nucleotide polymorphisms (SNPs) in the BACH2 gene are associated with Graves' disease (GD) in the Chinese Han population; however, the association did not achieve genome-wide significance levels. Recently, this association of BACH2 with GD was confirmed in Caucasians in the UK population, but fine mapping in this region has not yet been reported. Here, we provide a refined analysis of a 331-kb region in the BACH2 gene, which harbors 359 SNPs, using GWAS data from 1,442 GD patients and 1,468 controls. The SNPs rs2474619 and rs9344996 were implied as the independent variants associated with GD by forward and two-locus logistic regression analysis. We genotyped eight out of 10 tagSNPs with P < 1 × 10(-3) in 3,508 GD patients and 3,209 controls, the results also showed that rs2474619 was independently associated with GD in the combined population from GWAS and the second stage (P = 1.81 × 10(-5)). The rs2474619 and rs9344996 were further genotyped in the third stage cohorts, and rs2474619 showed evidence of association with GD at genome-wide significance levels in the combined population (P = 3.28 × 10(-8), odds ratio = 1.13). The association of rs9344996 with GD can be explained by its linkage to rs2474619 in the combined population. Our study clearly demonstrated that BACH2 is a susceptibility gene for GD in the Chinese Han population and further supported rs2474619, in intron 2 of BACH2, is the best association signal with GD. However, the mechanism by which BACH2 confers increased risk of GD requires further study.

Refined association of TSH receptor susceptibility locus to Graves' disease in the Chinese Han population.

BACKGROUND: Convincing evidence has demonstrated the association of TSH receptor (TSHR) with Graves' disease (GD) in the Chinese Han population. OBJECTIVE: The aim of this study was to identify the causal variants for GD in the region encompassing TSHR by a refining association study. DESIGN AND METHODS: GD patients (1536) and 1516 sex-matched controls were recruited in the first stage, and an additional 3832 GD patients and 3426 sex-matched controls were recruited in the replication stage. Genotyping was performed using Illumina Human660-Quad BeadChips or TaqMan single nucleotide polymorphism (SNP) Genotyping Assays and the Fluidigm EP1 platform. RESULTS: When the results of regression analysis for 74 genotyped SNPs and 922 imputed SNPs in the first-stage cohort were combined, rs179243 and rs3783949 were the probable susceptibility SNPs associated with GD in TSHR. Eleven SNPs, including rs179243 and rs3783949, were selected to further refine the association in the replication study. Finally, rs12101261 and rs179243 were confirmed as independent GD susceptibility variants in the replication and combined populations. Further, we also found that the rate of persistent TSHR autoantibody positivity (pTRAb+) was significantly higher in the GD patients with the susceptible genotypes rs12101261 or rs179243 than in the GD patients carrying the protective genotypes, after the GD patients had been treated for more than 1 year. CONCLUSIONS: These findings indicate that rs12101261 and rs179243 are the possible causal SNPs for GD susceptibility in the TSHR gene and could serve as genetic markers to predict the outcome of pTRAb+ in GD patients.

Whole-exome sequencing to identify novel somatic mutations in squamous cell lung cancers.

Squamous cell lung cancer is a major histotype of non-small cell lung cancer (NSCLC) that is distinct from lung adenocarcinoma. We used whole-exome sequencing to identify novel non-synonymous somatic mutations in squamous cell lung cancer. We identified 101 single-nucleotide variants (SNVs) including 77 non-synonymous SNVs (67 missense and 10 nonsense mutations) and 11 INDELs causing frameshifts. We also found four SNVs located within splicing sites. We verified 62 of the SNVs (51 missense, 10 nonsense and 1 splicing-site mutation) and 10 of the INDELs as somatic mutations in lung cancer tissue. Sixteen of the mutated genes were also mutated in at least one patient with a different type of lung cancer in the Catalogue of Somatic Mutation in Cancer (COSMIC) database. Four genes (LPHN2, TP53, MYH2 and TGM2) were mutated in approximately 10% of the samples in the COSMIC database. We identified two missense mutations in C10orf137 and MS4A3 that also occurred in other solid-tumor tissues in the COSMIC database. We found another somatic mutation in EP300 that was mutated in 4.2% of the 2,020 solid-tumor samples in the COSMIC database. Taken together, our results implicate TP53, EP300, LPHN2, C10orf137, MYH2, TGM2 and MS4A3 as potential driver genes of squamous cell lung cancer.

Robust evidence for five new Graves' disease risk loci from a staged genome-wide association analysis.

Graves' disease (GD), characterized by autoantibodies targeting antigens specifically expressed in thyroid tissues causing hyperthyroidism, is triggered by a combination of genetic and environmental factors. However, only a few loci for GD risk were confirmed in the various ethnic groups, and additional genetic determinants have to be detected. In this study, we carried out a three-stage study in 9529 patients with GD and 9984 controls to identify new risk loci for GD and found genome-wide significant associations in the overall populations for five novel susceptibility loci: the GPR174-ITM2A at Xq21.1, C1QTNF6-RAC2 at 22q12.3-13.1, SLAMF6 at 1q23.2, ABO at 9q34.2 and an intergenic region harboring two non-coding RNAs at 14q32.2 and one previous indefinite locus, TG at 8q24.22 (Pcombined < 5 × 10(-8)). The genotypes of corresponding variants at 14q32.2 and 8q24.22 were correlated with the expression levels of C14orf64 and a TG transcript skipping exon 46, respectively. This study increased the number of GD loci with compelling evidence and indicated that non-coding RNAs might be potentially involved in the pathogenesis of GD.