Andrade, Mariza de

1610 Background: Low dose computed tomography (LDCT) scans have reduced lung cancer deaths by 20.3% in high risk populations, although there is an unknown balance between the benefits and harms of LDCT scans as a screening tool. Our purpose was to compare health-related QOL issues among lung cancer patients who were initially detected by LDCT scans; 4 comparison groups included: lung cancer diagnosed by a screening chest X-ray, as an incidental finding from procedures taken for other medical reasons, or based on symptoms indicative for lung cancer and routinely diagnosed, and individuals who were LDCT screened but found no lung cancer (controls who participated in Mayo’s lung cancer CT screening trial). Methods: A total of 1,658 lung cancer patients (cared at Mayo Clinic) in the 4 groups (37, 151, 389, and 1081 respectively) and 488 controls were compared on following patient-reported outcomes (collected via validated tools): overall QOL, four symptoms (cough, pain, dyspnea, fatigue), mental/ physical/ emotional/ social/ spiritual QOL, and other concerns (e.g., family/ friends/ financial/ legal). A clinically significant deficit was defined as at least 10-points in difference (or <50 points) on a 0-100 scale. The rates of deficits were compared via Fisher’s exact tests and average QOL values via Kruskal-Wallis tests. Results: Overall QOL and individual symptoms were significantly worse (p<0.05) in all lung cancer groups than in controls, except for pain. LDCT-screened patients reported the greatest deficit among the 4 lung cancer groups in physical (41%), emotional (24%), social (38%), and spiritual QOL (24%); whereas chest X-ray detected patients had the least deficit in overall QOL (22%) and pain (32%). All 4 lung cancer groups experienced much worse fatigue (52-64%) than the controls (32%). Conclusions: Our preliminary results suggest that LDCT-screening detected lung cancer patients reported a different QOL profile from other lung cancer patients and non-lung cancer controls. The clinical course, smoking behavior, and QOL related health issues associated with LDCT screening for lung cancer warrant thorough investigation.

Introduction Severe obesity is a growing epidemic that causes significant morbidity and mortality, and is particularly difficult to reverse. Efficacious and cost-effective interventions are needed to combat this epidemic. This study hypothesized that obese people (body mass index (BMI) ≥35 kg/m 2 ) using a remote weight-loss program combining a mobile application, wireless scales, and low-calorie meal replacement would experience clinically significant weight loss. Methods This study was a retrospective observational analysis of 8275 individuals with a baseline BMI ≥35 kg/m 2 who used a remote weight-loss program combining mobile applications, frequent self-weighing, and calorie restriction via meal replacement for a minimum of 35 days. Weight changes were evaluated at multiple intervals (42, 60, 90, and 120 days), and weight loss was evaluated for all and for pre-specified subgroups based on demographic features and frequency of self-weighing. Results Mean weight loss at 42 days ( N = 6781) was 8.1 kg (margin of error (MOE) = 0.126 kg) with 73.6% of users experiencing >5% total body weight loss. Both men (9.1 kg; MOE = 0.172 kg; 7.9% from baseline) and women (7.1 kg; MOE = 0.179 kg; 7.2% from baseline) experienced significant weight loss. At the 120-day interval ( N = 2914), mean weight loss was 14 kg (MOE = 0.340 kg), 13% total body weight loss from baseline, and 82.3% of participants had lost >5% of their initial body weight. The decrease in body-fat percent correlated well with weight loss ( R = 0.92; p < 0.001). Conclusions In a large cohort of individuals with class II or III obesity, a remote weight-loss program combining mobile applications, daily self-weighing, and calorie restriction via meal replacement resulted in dramatic weight loss among subjects who were active users when evaluated through a retrospective observational analysis.

Introduction: Metabolic Syndrome represents a constellation of findings known to increase cardiovascular risk and all-cause mortality and is becoming more frequent in China. Smartphone applications may provide a cost-effective method to engage in weight loss and improve metabolic syndrome components. Methods: Prospective study of changes in weight and metabolic syndrome risk factors at 6 weeks and 6 months using a smartphone application utilizing a wireless scale, meal replacement biscuits and e-coaching in China. All participants had or were at high risk for metabolic syndrome and all interaction was online, without face to face contact. Results: 218 individuals ,55% female, participated. Median weight loss at 6 weeks and 6 months was 7.3kg and 8.4 kg (P<0.001) respectively. At baseline, 60% met a diagnosis of metabolic syndrome this dropped to 38% at 6 weeks (P<0.001) and stayed at 38% at 6 months (P<0.001). At 6 week the prevalence of all components of metabolic syndrome were improved (P<0.05) except HDL. Conclusions: A smartphone weight loss platform that utilizes a wireless scale, meal replacement biscuits and e-coaching was associated with significant weight loss and improvement in several metabolic syndrome components at 6 weeks and 6 months. This program may provide a scalable intervention against metabolic syndrome, especially in developing countries such as China.

Abstract Background: Lung cancer kills more people than any other cancer in the United States. In addition to environmental factors, lung cancer has genetic risk factors as well, though the genetic etiology is still not well understood. We have performed whole exome sequencing on 262 individuals from 28 extended families with a family history of lung cancer. Methods: Parametric genetic linkage analysis was performed on these samples using two distinct analyses—the lung cancer only (LCO) analysis, where only patients with lung cancer were coded as affected, and the all aggregated cancers (AAC) analysis, where other cancers seen in the pedigree were coded as affected. Results: The AAC analysis yielded a genome-wide significant result at rs61943670 in POLR3B at 12q23.3. POLR3B has been implicated somatically in lung cancer, but this germline finding is novel and is a significant expression quantitative trait locus in lung tissue. Interesting genome-wide suggestive haplotypes were also found within individual families, particularly near SSPO at 7p36.1 in one family and a large linked haplotype spanning 4q21.3-28.3 in a different family. The 4q haplotype contains potential causal rare variants in DSPP at 4q22.1 and PTPN13 at 4q21.3. Conclusions: Regions on 12q, 7p, and 4q are linked to increased cancer risk in highly aggregated lung cancer families, 12q across families and 7p and 4q within a single family. POLR3B, SSPO, DSPP, and PTPN13 are currently the best candidate genes. Impact: Functional work on these genes is planned for future studies and if confirmed would lead to potential biomarkers for risk in cancer.

Background The incidence of VTE in African-Americans (AAs) is similar to or higher than in Americans of European ancestry. However, the carrier frequencies of inherited thrombophilias common in whites (i.e., Factor V Leiden, Prothrombin G20210A) are very low in AAs, suggesting that other inherited thrombophilias may be associated with VTE in AAs. Objective To identify potentially novel non-hypothesis-driven single nucleotide polymorphisms (SNPs) associated with VTE in AAs. Methods We used the resources of the eMERGE Network to perform a GWAS of VTE in AAs. The eMERGE Network (funded by NHGRI) is comprised of nine sites each with DNA biobanks linked to electronic health records (EHRs). Approximately 39,206 unique DNA samples have been genotyped using either Affymetrix or Illumina genome-wide SNP arrays. Led by the Coordinating Center and eMERGE genomics workgroup, an imputation pipeline was developed for merging genomic data across the different SNP arrays used by the eMERGE sites, to maximize sample size and the power to detect associations. The imputation was performed using the 1000 Genomes Cosmopolitan reference panel which includes 1092 individuals and over 36 million SNPs. From our previously-identified cohort of Olmsted County, MN residents with incident or recurrent VTE, 1996-2005, we derived and validated an EHR-driven phenotype extraction algorithm that leveraged structured data based on ICD-9-CM codes and unstructured data from clinical notes via natural language processing to identify VTE cases and controls with 100% and 94% positive and negative predictive values, respectively. We tested for an association between each SNP and VTE among AAs using unconditional logistic regression, adjusting for age, sex and eMERGE site. Results Among 294 AA VTE cases and 3,661 AA controls (total n=3,955; females, n=2,512), the Factor V Leiden (F5 rs6025) was not analyzed due to a very low minor allele frequency (MAF=0.0036). The prothrombin G20210A (F2 rs1799963) and ABO blood type O (ABO rs8176719) SNPs were not genotyped in any of the arrays and could not be imputed. Among SNPs with an imputation score >0.8, the most significant SNPs associated with VTE were ITPR3 (inositol 1,4,5-triphosphate receptor type 3) rs2229637 (OR=1.65; p=3.61E-07; MAF=0.19) and CLEC7A (C-type lectin domain family 7, member A) rs59819090 (OR=2.16; p=1.06E-06; MAF=0.056). ITPR3 SNPs have been associated with coronary artery aneurysm in Kawasaki disease, type 1 diabetes mellitus and other autoimmune disorders. CLEC7A rs59819090 encodes for a serine82 to leucine nonsynonymous amino acid change in dectin-1. Dectin-1 is a transmembrane innate immune pattern recognition receptor on myeloid cells that upon binding it’s agonist, β-glucans, stimulates cytokine production and the respiratory burst, a prerequisite for formation of neutrophil extracelluar traps (NETs). NETs have been associated with VTE (van Montfoort, et al. Ateriosclero Thromb Vasc Biol 2013;33:147-51). Dectin-1 serine82 is not evolutionarily conserved and the serine82 to leucine amino acid change is predicted to be tolerated, with a moderate effect on protein function. Conclusions ITPR3 rs2229637 and CLAC7A rs59819090 may be associated with VTE in African-Americans. These observations require replication and functional studies toward understanding how they may lie on the causal pathway to VTE. Disclosures: No relevant conflicts of interest to declare.

Abstract Abstract 709 Background: A recent analysis of merged genome-wide and candidate gene genotypes in VTE cases and controls identified multiple tag SNPs that were strongly associated with VTE. Objective: To identify rare and/or novel functional variants by sequencing the implicated genes. Methods: Cases (n=1488) were Mayo Clinic European-American patients of non-Hispanic ancestry with objectively-diagnosed VTE in the absence of active cancer, venous catheter or antiphospholipid antibodies. Controls (n=1439) were Mayo Clinic outpatients without VTE who were frequency-matched on case age, gender, race, MI/stroke status and state of residence. For this analysis, we selected a subset of these cases and controls for sequencing to take advantage of the joint configuration of two ABO SNPs of primary interest, rs8176719 (ABO exon 6 deletion determining type O blood group) and rs2519093 (ABO intron 1 tag SNP), which were previously shown to be strongly associated with VTE (p=5.7E-12 and p=3.0E-16, respectively). We randomly sampled 82 cases and 14 controls within 3 of the 9 potential allele frequency cells (Figure). The rs8176719 alleles are -−/−- (double deletion is the common allele), –/G, and G/G (the rare allele). The rs2519093 alleles are GG (G is the common allele), AG, and AA (A is the rare allele). For each SNP, the genotypes are represented as 0, 1, or 2 copies of the minor allele. We represented the joint allelic configuration of the two SNPs with the number of copies of the rs8176719 given first as 0/0 (both with 0 copies of the minor allele), 0/1, 0/2 (0 copies of the rs8176719 SNP), 1/0 (1 copy of the rare rs8176719 SNP), 1/1/, and 1/2, and 2/0 (2 copies of the rare allele for the rs8176719 SNP and 0 copies of the rare allele of the rs2519093 SNP), 2/1, and 2/2. From the Figure one observes discrepancies between cases and controls at the 0/0, 1/1 and 2/2 combinations. We randomly sampled from these three combinations, taking one third of the case series. For each SNP, we had 28 cases with 0/0 copies of the rare allele, 27 cases with 1/1 copies of the rare allele; and 27 cases with the combination of 2/2 copies of the rare allele. We compared these 82 cases with 14 controls that do not have any of these combinations. Sixteen genes were selected for deep sequencing, including 5 genes harboring SNPs significantly associated with VTE (F5, SLC19A2, ABO, NME7, ATP1B1), 10 genes with SNPs marginally associated with VTE (C1orf114, KLKB1, SELP, F11, SCUBE1, PRKCB1, CD44, ITPR1, GFRA1, BLZF1), and CYP4V2 which reportedly confounds F11 and KLKB1. Agilent SureSelect probes were designed to capture and enrich the ∼2 Mb genomic regions of these 16 genes. Samples were multiplexed (12-plex) and sequenced using Illumina HiSeq 2000. The sequence reads were aligned to the human genome build 36 using Burrows-Wheeler Aligner, and the single nucleotide variants (SNVs) and small INDELs were called using SNVMix and GATK, respectively. For this analysis, novel ABO SNVs were tested for an association with VTE using age-, sex-adjusted logistic regression and Fisher's Exact Test. Results: 98% of the targeted regions were sequenced with > 20X coverage. On average, ∼2500 SNVs and ∼200 INDELs were detected in each sample. Fifteen novel SNVs in intron 6 and 3' of the ABO gene were associated with VTE (p<E-06) and belonged to 3 distinctive LD blocks; none were in LD with the coding or tag ABO SNPs (rs8176719; rs2519093). SNVs inside the middle LD block at the 3' of ABO are located within an enhancer and promoter histone marked with putative transcription factor binding sites. In addition, strong evidence from both ENCODE and dbEST support the middle LD block as lying within a novel transcript, probably an extension of the 3' of ABO. In addition, we discovered a novel, significant, protective, frame-shifting single base (G) deletion at ABO chr9:135120877. Conclusion: Novel ABO functional variants that are associated with VTE were identified by deep sequencing. Disclosures: Heit: Daiichi Sankyo: Consultancy, Honoraria.

Abstract Abstract 1238 Background: SNPs within genes encoding factor XI (F11), fibrinogen genes (FGA, FGG) and other candidate genes within the procoagulant, anticoagulant, fibrinolytic, innate immunity and endocrine pathways have been reported as associated with VTE. However, the independent risk of VTE associated with many of these SNPs after controlling for factor V Leiden, Prothrombin G20210A and ABO blood group non-O carrier status is uncertain. Objective: To replicate candidate gene SNPs previously reported as associated with VTE. Methods: As part of a large replication study, we included 17 SNPs previously reported as associated with VTE in a custom Illumina Golden gate (total n=1093 SNPs) genotyping array. We genotyped 1270 non-Hispanic adults of European ancestry with objectively-diagnosed VTE (cases; no cancer, venous catheter or antiphospholipid antibodies) and 1302 controls (frequency-matched on case age, gender, race, MI/stroke status). Genotyping results from high-quality control DNA (SNP call rate ≥ 95%) was used to generate a cluster algorithm. The primary outcome was VTE status, a binary measure. The covariates were age at interview or blood sample collection, sex, stroke and/or MI status, and state of residence. To adjust for population stratification, we performed the multidimensional scaling (MDS) analysis option in PLINK v 1.07 to identify outliers in our population using the ancestry informative markers. We tested for an association between each SNP and VTE using unconditional logistic regression, adjusting for age, sex, stroke/MI status, state of residence and ABO rs514659 (in high linkage disequilibrium with non-O blood type). The analyses were corrected for multiple comparisons using an extension of false discovery rates. The false discovery rate (reported as a Q-value) is an analogue measure of the p-value that takes into account the number of statistical tests and estimates the expected proportion of false positive tests incurred when a particular SNP is significant. All analyses were performed using PLINK v 1.07. Results: MDS gave no evidence of population stratification. Genotyping was unsuccessful for two of the 17 SNPs. We found significant associations between VTE and SNPs in F11, FGG, TC2D and FGA (Table). However, the false discovery rates for all significant SNPs except F11 rs3756008 were >0.05, suggesting that the observed associations were likely falsely positive due to multiple comparisons. Even at a false discovery rate of Q-value=0.0099, one would expect ∼13 SNPs (0.0099 × 1302 SNPs) to be falsely associated with VTE due to multiple comparisons. Consequently, even our observed association between F11 rs3756008 and VTE remains tentative. Conclusions: We were unable to replicate reported associations between 15 SNPs and VTE. Our results emphasize the necessity of replication studies in different populations to confirm reported associations of SNPs with VTE. Disclosures: Heit: Daiichi Sankyo: Consultancy, Honoraria.

Abstract Abstract 1242 Background: While bivariate interactions between Factor V Leiden, prothrombin G20201A and hereditary deficiency of antithrombin, protein C and protein S compound VTE risk, whether other gene-gene interactions are associated with VTE is largely unknown. Objective: To test novel gene-gene interactions for an association with VTE. Methods: Genome-wide scan (Illumina 660Q [557,112 SNPs]) and candidate gene (12,551 SNPs in 764 genes within the anticoagulant, procoagulant, fibrinolytic and innate immunity pathways) genotypes from 1270 non-Hispanic adults of European ancestry with objectively-diagnosed VTE (cases; no cancer, venous catheter or antiphospholipid antibodies) and 1302 controls (frequency-matched on case age, gender, race, MI/stroke status) were merged and imputed to about 2.5 million SNPs with MACH using HapMap Phase 2 (60 CEU). We tested all pairwise SNP-SNP interactions using a 2-stage procedure (fast epistasis in the first stage and age, gender, MI/stroke status and state of residence adjusted analysis using logistic regression in the second stage). Results: The mean ± standard deviation case and control ages were 54.4 ± 16.2 and 55.6 ± 15.8 years, respectively and 51% were female. Among 2.299×10^12 pairwise interactions tested, 900 million reached a p-value<E-06 and were be tested in the second stage. Of these, 2417, 278 and 22 SNP-SNP interactions reached p-values ≤1E-9, 1E–10, and1E-11 respectively; 121 SNP-SNP interactions reached the Bonferroni statistical significance threshold (≤5.555659E-11). The gene-gene pairwise interactions with p-values≤1E-11 were: (1) chromosome 3 open reading frame 70 (C3orf70) – short-chain dehydrogenase/reductase (SDR family) member 4 (DHRS4), (2) 1-acylglycerol-3-phosphate O-acyltransferase 4 (AGPAT4) - NIMA (never in mitosis gene a)-related kinase 6 (NEK6), and (3) TatD DNase domain containing 2 (TATDN2) – WAS protein family, member 3 (WASF3). No gene function data are available for C3orf70. DHRS4 encodes for a 3-ketosteroid reductase important for active steroid hormone regulation, AGPAT4 is important for signal transduction and lipid biosynthesis, NEK6 encodes for Nek6 which downregulates p53-induced cancer cell senescence, TATDN2 encodes a deoxyribonuclease, WASF3 encodes for a member of the Wiskott-Aldrich syndrome protein family which are key regulators of signal transduction and cytoskeletal reorganization in hematopoietic cells. Conclusion: Pairwise gene-gene interaction analyses suggest potential associations between VTE and novel genes for future testing in replication studies. Disclosures: Heit: Daiichi Sankyo: Consultancy, Honoraria.

Abstract Abstract 2295 Background: While interaction between Factor V Leiden and other VTE risk exposures compound VTE risk, whether other gene-environment interactions are associated with VTE is largely unknown. Objective: To test novel gene-environment interactions for an association with VTE. Methods: Genome-wide scan (Illumina 660Q [557,112 SNPs]) and candidate gene (12,551 SNPs in 764 genes within the anticoagulant, procoagulant, fibrinolytic and innate immunity pathways) genotypes from 1270 non-Hispanic adults of European ancestry with objectively-diagnosed VTE (cases; no cancer, venous catheter or antiphospholipid antibodies) and 1302 controls (frequency-matched on case age, gender, race, MI/stroke status) were merged and imputed to 2.5 million SNPs with MACH using HapMap Phase 2 (60 CEU). We tested pairwise interactions between these SNPS and each of 7 exposures (“stress” [defined as ever hospitalized, ever surgery, ever trauma, ever transfemoral procedure or ever leg paresis], ever surgery, ever neurosurgery, ever orthopedic surgery; and among women, ever pregnant, ≥3 pregnancies and ever oral contraceptives/hormone therapy), adjusted for age, gender, MI/stroke status and state of residence, using logistic regression. Results: The mean ± standard deviation case and control ages were 54.4 ± 16.2 and 55.6 ± 15.8 years, respectively and 51% were female. Among the 7 models, a significant interaction was found between stress and A2BP1 (rs735919, OR=0.2, p=2.8E-07), ever surgery and ABCG8 (rs6709904, OR=0.27, p=2.1E-07), orthopedic surgery and SLC24A3 (rs6081599, OR=0.53, p=2.8E-07), neurosurgery and TBC1D4 (rs17064485, OR=8.5, p=3.5E-07), every pregnant and MICALCL (rs11022321, OR=0.17, p=8.3E-07), >3 pregnancies and KLRB1 (rs2536929, OR=0.31, p=1.1E-07) and ever oral contraceptives/hormone therapy and LINGO2 (rs10969259, OR=2.02, p=4.9E-07). A2BP1 plays a role in RBC shape and size, ABCG8 is associated with severe hypercholesterolemia, SLC24A3 is important for intracellular calcium homeostasis, TBC1D4 participates in cell signaling in CD4+/CD45RO+ memory T-cells, MICALCL is an extracellular signal-regulated kinase 2-binding protein, KLRB1 regulates natural killer cell function, and LINGO2 encodes a leucine-rich repeat signaling protein. Our observed interaction of VTE risk factors with these genes to compound the risk of VTE requires replication in future studies. If confirmed, functional studies will be important to determine the roles of these gene products in the causal pathway to VTE. Conclusion: VTE risk factors and SNPs within several novel genes may interact to compound the risk of VTE. Disclosures: Heit: Daiichi Sankyo: Consultancy, Honoraria.

Abstract Abstract 480 Background: While interaction between Factor V Leiden and other VTE risk exposures (i.e., oral contraceptives, hormone therapy, pregnancy, cancer, minor trauma) compound VTE risk, whether other gene-environment interactions are associated with VTE is largely unknown. Objective: To test gene-environment interactions for an association with VTE. Methods: Cases (n=1488) were Mayo Clinic European-American patients of non-Hispanic ancestry with objectively-diagnosed VTE in the absence of active cancer, venous catheter or antiphospholipid antibodies. Controls (n=1439) were Mayo Clinic outpatients without VTE who were frequency-matched on case age, gender, race, MI/stroke status and state of residence. We selected candidate genes relevant to the anticoagulant, procoagulant, fibrinolytic and innate immunity pathways, focusing on platelet, monocyte, neutrophil and endothelial cell agonists, receptors, ligands, signal transduction and adhesion molecules, granule contents and effectors; plasma proteases and inhibitors; matrix metalloproteases; inflammatory cytokines and receptors; estrogen, progesterone and androgen receptors; co-regulators and enzymes related to estrogen metabolism; important enzymes for catechol, homocysteine, thromboxane A2 and prostacyclin biosynthesis and metabolism; and HMG CoA reductase. For these genes (n=750), we selected all non-synonymous coding single nucleotide polymorphisms (SNPs) with minor allele frequency ≥0.5%; the remaining SNPs were selected using an LD tagging algorithm (Carlson et al. AJHG 2004); 479 ancestry-informative markers were also included (total n=13,241 SNPs). Leukocyte genomic DNA was genotyped using a custom Illumina Infinium iSelect platform, including appropriate controls. We tested all pairwise interactions between 12,483 SNPs (12296 autosomal, 187 chromosome X) that passed quality control and each of seven exposures (ever “stress”[defined as ever hospitalized, ever surgery, ever trauma, ever transfemoral procedure or ever leg paresis], ever surgery, ever neurosurgery, ever orthopedic surgery; and among women, ever pregnant, ≥3 pregnancies and ever oral contraceptives/hormone therapy), adjusted for age, gender, MI/stroke status and state of residence, using logistic regression. False discovery rates (q-value) were calculated to estimate the expected fraction of false positive associations due to multiple statistical tests. Results: The mean ± SD case and control ages were 55.3 ± 16.4 and 56.5 ± 15.9 years, respectively, and 51% were female. Analysis of ancestry-informative markers revealed no evidence of population stratification. Among the seven models, a significant interaction (q≤0.05) was found between ever surgery and two genes located on chromosome 1: USF1 (rs2516840, OR=2.1, p=2.0E-06; q=0.02), and F11R (rs790055 & rs790056; OR∼2.29, p∼4E-06; q=0.02); the two F11R SNPs are in complete linkage dysequilibrium (LD), and the USF1 SNP is in moderate LD with the F11R SNPs (r2∼0.6). USF1 encodes for upstream transcription factor 1 which regulates many genes involved in lipid and glucose homeostasis. F11R encodes for the platelet F11 receptor (JAM [junctional adhesion molecule], a cell adhesion molecule important for platelet adhesion to cytokine-stimulated endothelial cells. Conclusion: Interaction between surgery and SNPs within USF1 and F11R are associated with significantly increased risks for VTE. These potential associations require confirmation in future replication studies. Disclosures: No relevant conflicts of interest to declare.