Systematic meta-analyses, field synopsis and global assessment of the evidence of genetic association studies in colorectal cancer.

OBJECTIVE: To provide an understanding of the role of common genetic variations in colorectal cancer (CRC) risk, we report an updated field synopsis and comprehensive assessment of evidence to catalogue all genetic markers for CRC (CRCgene2). DESIGN: We included 869 publications after parallel literature review and extracted data for 1063 polymorphisms in 303 different genes. Meta-analyses were performed for 308 single nucleotide polymorphisms (SNPs) in 158 different genes with at least three independent studies available for analysis. Scottish, Canadian and Spanish data from genome-wide association studies (GWASs) were incorporated for the meta-analyses of 132 SNPs. To assess and classify the credibility of the associations, we applied the Venice criteria and Bayesian False-Discovery Probability (BFDP). Genetic associations classified as 'positive' and 'less-credible positive' were further validated in three large GWAS consortia conducted in populations of European origin. RESULTS: We initially identified 18 independent variants at 16 loci that were classified as 'positive' polymorphisms for their highly credible associations with CRC risk and 59 variants at 49 loci that were classified as 'less-credible positive' SNPs; 72.2% of the 'positive' SNPs were successfully replicated in three large GWASs and the ones that were not replicated were downgraded to 'less-credible' positive (reducing the 'positive' variants to 14 at 11 loci). For the remaining 231 variants, which were previously reported, our meta-analyses found no evidence to support their associations with CRC risk. CONCLUSION: The CRCgene2 database provides an updated list of genetic variants related to CRC risk by using harmonised methods to assess their credibility.

High-Resolution Ecological Niche Modeling of Ixodes scapularis Ticks Based on Passive Surveillance Data at the Northern Frontier of Lyme Disease Emergence in North America.

BACKGROUND: Lyme disease (LD) is a bacterial infection transmitted by the black-legged tick (Ixodes scapularis) in eastern North America. It is an emerging disease in Canada due to the expanding range of its tick vector. Environmental risk maps for LD, based on the distribution of the black-legged tick, have focused on coarse determinants such as climate. However, climatic factors vary little within individual health units, the level at which local public health decision-making takes place. We hypothesize that high-resolution environmental data and routinely collected passive surveillance data can be used to develop valid models for tick occurrence and provide insight into ecological processes affecting tick presence at fine scales. METHODS: We used a maximum entropy algorithm (MaxEnt) to build a habitat suitability model for I. scapularis in Ottawa, Ontario, Canada using georeferenced occurrence points from passive surveillance data collected between 2013 and 2016 and high-resolution land cover and elevation data. We evaluated our model using an independent tick presence/absence dataset collected through active surveillance at 17 field sites during the summer of 2017. RESULTS: Our model showed a good ability to discriminate positive sites from negative sites for tick presence (AUC = 0.878 ± 0.019, classification accuracy = 0.835 ± 0.020). Heavily forested suburban and rural areas in the west and southwest of Ottawa had higher predicted suitability than the more agricultural eastern areas. CONCLUSIONS: This study demonstrates the value of passive surveillance data to model local-scale environmental risk for the tick vector of LD at sites of interest to public health. Given the rising incidence of LD and other emerging vector-borne diseases in Canada, our findings support the ongoing collection of these data and collaboration with researchers to provide a timely and accurate portrait of evolving public health risk.

Waterpipe smoking and cancer: systematic review and meta-analysis.

OBJECTIVE: Although accumulating evidence suggests harmful effects of waterpipe smoking, there is limited information about its direct association with chronic diseases, notably cancer. We provide an up-to-date systematic review and meta-analysis of the association between waterpipe smoking and cancer. DATA SOURCES: Systematic search of articles indexed in main biomedical databases: Pubmed, EmBase, Google Scholar and Web of Science, published between 1962 and September 2014. Search keywords included a combination of waterpipe or hookah, sheesha, nargile, hubble-bubble, goza or gaylan, and cancer. STUDY SELECTION: Focus on observational studies (cohort, case-control, cross-sectional) that evaluated the association between waterpipe smoking and cancer. Studies with mixed exposures excluded. DATA EXTRACTION: Two investigators independently extracted data and reached consensus on all items. DATA SYNTHESIS: 13 case-control studies met the inclusion criteria and were considered for meta-analysis. The methodological quality of included studies was assessed using the Newcastle-Ottawa Scale (NOS). Meta-analysis revealed a positive association between waterpipe smoking and lung cancer (OR=4.58 (2.61 to 8.03); I2=44.67%), and oesophageal cancer (OR=3.63 (1.39 to 9.44); I2 =94.49%). The majority of studies had a NOS score of 5-6 or 7, indicating 'fair' or 'good' quality, respectively. CONCLUSIONS: Our findings support a positive association between waterpipe smoking and cancer risk. However, high-quality studies with standardised exposure measurements are needed to clarify the contribution of waterpipe smoking to chronic diseases. More investments in initiatives for surveillance, intervention and regulatory policy for waterpipe smoking are urgently warranted.

Systematic meta-analyses and field synopsis of genetic association studies in colorectal adenomas.

BACKGROUND: Low penetrance genetic variants, primarily single nucleotide polymorphisms, have substantial influence on colorectal cancer (CRC) susceptibility. Most CRCs develop from colorectal adenomas (CRA). Here we report the first comprehensive field synopsis that catalogues all genetic association studies on CRA, with a parallel online database [http://www.chs.med.ed.ac.uk/CRAgene/]. METHODS: We performed a systematic review, reviewing 9750 titles, and then extracted data from 130 publications reporting on 181 polymorphisms in 74 genes. We conducted meta-analyses to derive summary effect estimates for 37 polymorphisms in 26 genes. We applied the Venice criteria and Bayesian False Discovery Probability (BFDP) to assess the levels of the credibility of associations. RESULTS: We considered the association with the rs6983267 variant at 8q24 as 'highly credible', reaching genome-wide statistical significance in at least one meta-analysis model. We identified 'less credible' associations (higher heterogeneity, lower statistical power, BFDP > 0.02) with a further four variants of four independent genes: MTHFR c.677C>T p.A222V (rs1801133), TP53 c.215C>G p.R72P (rs1042522), NQO1 c.559C>T p.P187S (rs1800566), and NAT1 alleles imputed as fast acetylator genotypes. For the remaining 32 variants of 22 genes for which positive associations with CRA risk have been previously reported, the meta-analyses revealed no credible evidence to support these as true associations. CONCLUSIONS: The limited number of credible associations between low penetrance genetic variants and CRA reflects the lower volume of evidence and associated lack of statistical power to detect associations of the magnitude typically observed for genetic variants and chronic diseases. The CRA gene database provides context for CRA genetic association data and will help inform future research directions.

Hypermethylation of the 5' CpG island of the p14ARF flanking exon 1ß in human colorectal cancer displaying a restricted pattern of p53 overexpression concomitant with increased MDM2 expression.

BACKGROUND: It has been suggested that inactivation of p14ARF, a tumor suppressor central to regulating p53 protein stability through interaction with the MDM2 oncoprotein, abrogates p53 activity in human tumors retaining the wild-type TP53 gene. Differences in expression of tumor suppressor genes are frequently associated with cancer. We previously reported on a pattern of restricted p53 immunohistochemical overexpression significantly associated with microsatellite instability (MSI), low TP53 mutation frequency, and MDM2 overexpression in colorectal cancers (CRCs). In this study, we investigated whether p14ARF alterations could be a mechanism for disabling the p53 pathway in this subgroup of CRCs. RESULTS: Detailed maps of the alterations in the p14ARF gene were determined in a cohort of 98 CRCs to detect both nucleotide and copy-number changes. Methylation-specific PCR combined with bisulfite sequencing was used to evaluate the prevalence and distribution of p14ARF methylation. p14ARF alterations were then correlated with MSI status, TP53 mutations, and immunohistochemical expression of p53 and MDM2. The frequency of p14ARF mutations was extremely low (1/98; 1%), whereas coexistence of methylated and unmethylated alleles in both tumors and normal colon mucosa was common (91/98; 93%). Only seven of ninety-eight tumors (7%) had a distinct pattern of methylation compared with normal colon mucosa. Evaluation of the prevalence and distribution of p14ARF promoter methylation in a region containing 27 CpG sites in 35 patients showed a range of methylated CpG sites in tumors (0 to 25 (95% CI 1 to 13) versus 0 to 17 (95% CI 0 to 2)) in adjacent colon mucosa (P = 0.004). Hypermethylation of the p14ARF promoter was significantly correlated with the restricted p53 overexpression pattern (P = 0.03), and MDM2 overexpression (P = 0.02), independently of MSI phenotype. Although no significant correlation between p14ARF methylation and TP53 mutational status was seen (P = 0.23), methylation involving the proximal CpG sites within the 5' CpG flanking exon 1ß was present more frequently in tumors with restricted p53 overexpression than in those with diffuse p53 overexpression (range of methylated clones 17 to 36% (95% CI 24 to 36%) versus range 0 to 3% (95% CI 0 to 3%), P = 0. 0003). CONCLUSION: p14ARF epigenetic silencing may represent an important deregulating mechanism of the p53-MDM2-p14ARF pathway in CRCs exhibiting a restricted p53 overexpression pattern.

Distinctive patterns of p53 protein expression and microsatellite instability in human colorectal cancer.

Although evidence suggests an inverse relationship between microsatellite instability and p53 alterations in colorectal cancer, no study has thoroughly examined the use of p53 immunohistochemistry in phenotyping colorectal cancers. We investigated the value of p53 immunohistochemistry in microsatellite instability-positive colorectal cancers prescreening and attempted to clarify the relationship between DNA mismatch repair system and p53 pathway. In a series of 104 consecutive colorectal cancers, we performed p53 immunohistochemistry, TP53 mutational analysis, DNA mismatch repair system efficiency evaluation (DNA mismatch repair system immunohistochemistry, microsatellite instability status, MLH1/MSH2 germ line, and BRAF, murine double minute 2, and p21 immunohistochemistry. Microsatellite instability high was observed in 25 of 104 colorectal cancers, with DNA mismatch repair system protein loss (24/25) and germ line (8/25) or BRAF mutations (8/25). p53 immunohistochemistry revealed 3 distinct patterns of expression: complete negative immunostaining associated with truncating TP53 mutations (P < .0001), diffuse overexpression associated with missense TP53 mutations (P < .0001), and restricted overexpression characterized by a limited number of homogenously scattered strongly positive tumor cells in 36.5% of colorectal cancers. This latest pattern was associated with wild-type TP53 and microsatellite instability high colorectal cancers (P < .0001) including all Lynch tumors (8/8), but its presence among 22% of DNA mismatch repair system-competent colorectal cancers decreased its positive predictive value (55.2% [95% confidence interval, 45%-65%]). It was also correlated with murine double minute 2 overexpression (P < .0001) and inversely with p21 loss (P = .0002), independently of microsatellite instability status. In conclusion, a restricted pattern of p53 overexpression is preferentially associated with microsatellite instability high phenotype and could, therefore, be of clinical use as signal for microsatellite instability analysis in a large-scale tumor screening. Its association with concomitant murine double minute 2 overexpression suggests an alternative mechanism of p53 pathway deregulation.

High risk for neoplastic transformation of endometriosis in a carrier of Lynch syndrome.

Lynch syndrome is an autosomal dominant cancer-susceptibility disorder caused by mutations in DNA mismatch repair genes. Women with Lynch syndrome have an increased lifetime risk for endometrial and ovarian cancers. While there is evidence of efficacy for prophylactic surgery, no standard recommendations have been developed to support screening for premalignant endometrial and ovarian epithelial lesions in high-risk women. Here, we report a case of a healthy woman carrying a germline mutation in MLH1 gene with endometrial intra-epithelial neoplasia and ovarian endometriotic lesions exhibiting a loss of MLH1 protein expression. This case report illustrates the malignant potential of endometriosis, and highlights the need for a meticulous management of gynecologic premalignant precursor lesions in reducing cancer risk among related Lynch syndrome women.