Epigenetic regulation of Amphiregulin and Epiregulin in colorectal cancer.

Expression of the epidermal growth factor ligands amphiregulin (AREG) and epiregulin (EREG) is positively correlated with a response to EGFR-targeted therapies in colorectal cancer. Gene-body methylation sites, which show a strong inverse correlation with AREG and EREG gene expression, were identified in cell lines using targeted 454 FLX-bisulfite sequencing and SIRPH analyses for AREG/EREG promoters and intragenic CpGs. Upon treatment of colorectal cancer cells with 5-aza-2'-desoxycytidine, methylation decreases at specific intragenic CpGs accompanied by upregulation of AREG and EREG gene expression. The same AREG gene-body methylation was also found in human colorectal cancer samples and is independent of KRAS and NRAS mutations. Methylation is specifically decreased in the tumor epithelial compartment as compared to stromal tissue and normal epithelium. Investigation of a promoter/enhancer function of the AREG exon 2 region revealed a potential promoter function in reverse orientation. Retrospective comparison of the predictive power of AREG gene-body methylation versus AREG gene expression using samples from colorectal cancer patients treated with anti-EGFR inhibitors with complete clinical follow-up revealed that AREG expression is superior to AREG gene methylation. AREG and EREG genes undergo a complex regulation involving both intragenic methylation and promoter-dependent control.

The non-human primate kidney transcriptome in fetal development.

BACKGROUND: Little is known about the repertoire of non-human primate kidney genes expressed throughout development. The present work establishes an understanding of the primate renal transcriptome during fetal development in the context of renal maturation. METHODS: The baboon kidney transcriptome was characterized at 60-day gestation (DG), 90 DG, 125 DG, 140 DG, 160 DG and adulthood (6-12 years) using gene arrays and validated by QRT-PCR. Pathway and cluster analyses were used to characterize gene expression in the context of biological pathways. RESULTS: Pathway analysis indicated activation of pathways not previously reported as relevant to kidney development. Cluster analysis also revealed gene splice variants with discordant expression profiles during development. CONCLUSIONS: This study provides the first detailed genetic analysis of the developing primate kidney, and our findings of discordant expression of gene splice variants suggest that gene arrays likely provide a simplified view and demonstrate the need to study the fetal renal proteome.

Long-Term Functional Outcomes in Pediatric Head and Neck Cancer Patients: A Systematic Review.

OBJECTIVE: The multimodal treatments for pediatric head and neck (H&N) malignancies can have significant long-term functional consequences for growing patients. This systematic review aims to analyze the current knowledge of functional outcomes for pediatric H&N cancer survivors. DATA SOURCES: PubMed, Embase, Web of Science. REVIEW METHODS: Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines were followed, and 1356 papers were reviewed by 3 team members with conflict resolution by a senior member. RESULTS: Fourteen studies were included. Nine of 14 (64%) papers reported issues with swallowing, characterized as either dysphagia, odynophagia, oropharyngeal fibrosis, esophageal stenosis, xerostomia, trismus, or general issues with the throat and mouth. Six of 14 papers noted nutritional and feeding deficiencies, and 5 of 14 additionally noted issues with speech and voice changes. Four of 14 (29%) reported hearing impairments and/or loss. A majority of papers (9/14) reported long-term functional characteristics as a secondary outcome. Three of 14 (21%) reported a quality of life (QoL) measure. Heterogeneity in methodology and reporting precluded analysis of any relationship between treatment type and functional outcomes. Recommendations include integration of objective measures of feeding support and swallowing, as well as regular measurements of function and QoL parameters during treatment to better understand the evolution of QoL and function throughout care. CONCLUSION: Relatively few studies focus on functional outcomes following the treatment of pediatric H&N cancer. Swallowing difficulty is the most frequently reported deficit, but objective data is rarely reported. Standardization of functional outcome assessment could improve the quality of evidence for pediatric patients treated for H&N cancer.

Mutation-specific effects of NRAS oncogenes in colorectal cancer cells.

In colorectal cancer (CRC), the prevalence of NRAS mutations (5-9%) is inferior to that of KRAS mutations (40-50%). NRAS mutations feature lately during tumour progression and drive resistance to anti-EGFR therapy in KRAS wild-type tumours. To elucidate specific functions of NRAS mutations in CRC, we expressed doxycycline-inducible G12D and Q61K mutations in the CRC cell line Caco-2. A focused phospho-proteome analysis based on the Bio-Plex platform, which interrogated the activity of MAPK, PI3K, mTOR, STAT, p38, JNK and ATF2, did not reveal significant differences between Caco-2 cells expressing NRASG12D, NRASQ61K and KRASG12V. However, phenotypic read-outs were different. The NRAS Q61K mutation promoted anchorage-independent proliferation and tumorigenicity, similar to features driven by canonical KRAS mutations. In contrast, expression of NRASG12D resulted in reduced proliferation and apoptosis. At the transcriptome level, we saw upregulation of cytokines and chemokines. IL1A, IL11, CXCL8 (IL-8) and CCL20 exhibited enhanced secretion into the culture medium. In addition, RNA sequencing results indicated activation of the IL1-, JAK/STAT-, NFκB- and TNFα signalling pathways. These results form the basis for an NRASG12D-driven inflammatory phenotype in CRC.

Generation after generation: exploring the psychological impact of providing genetic services through a cascading approach.

PURPOSE: The provision of genetic services often occurs in a cascading fashion within families experiencing inherited diseases. This study examines whether previous family experiences with genetic services influences levels of psychological well-being of family members receiving services later. METHODS: Two hundred ninety-seven persons from 38 families with Lynch syndrome completed questionnaires before receiving genetic services. Baseline levels of test-related distress, depressive symptoms, and cancer worries were assessed in relationship to the (1) amount of time elapsed since services were provided to the index case and (2) generation of the family member relative to the index case. RESULTS: Family members in the same generation as the index case experienced significant increases in test-related distress (P = 0.003) and cancer worry (P = 0.001) with increasing time between receipt of genetic test results by the index case and provision of services to family members. Change in the number of depressive symptoms was not significant (P = 0.17). CONCLUSION: The provision of genetic services through a cascading approach significantly increases distress and worry among family members within the same generation as the index case who receive services at increasingly distant time intervals. Additional research is needed to explore social influences after the introduction of genetic services.

Characteristics of health information gatherers, disseminators, and blockers within families at risk of hereditary cancer: implications for family health communication interventions.

OBJECTIVES: Given the importance of the dissemination of accurate family history to assess disease risk, we characterized the gatherers, disseminators, and blockers of health information within families at high genetic risk of cancer. METHODS: A total of 5466 personal network members of 183 female participants of the Breast Imaging Study from 124 families with known mutations in the BRCA1/2 genes (associated with high risk of breast, ovarian, and other types of cancer) were identified by using the Colored Eco-Genetic Relationship Map (CEGRM). Hierarchical nonlinear models were fitted to characterize information gatherers, disseminators, and blockers. RESULTS: Gatherers of information were more often female (P<.001), parents (P<.001), and emotional support providers (P<.001). Disseminators were more likely female first- and second-degree relatives (both P<.001), family members in the older or same generation as the participant (P<.001), those with a cancer history (P<.001), and providers of emotional (P<.001) or tangible support (P<.001). Blockers tended to be spouses or partners (P<.001) and male, first-degree relatives (P<.001). CONCLUSIONS: Our results provide insight into which family members may, within a family-based intervention, effectively gather family risk information, disseminate information, and encourage discussions regarding shared family risk.

Comparative Biocompatibility and Osteogenic Potential of Two Bioceramic Sealers.

INTRODUCTION: Endodontic sealers have traditionally been used to seal dentinal tubules, creating a homogenous interface between the obturation material and the dentinal walls. However, bioceramic sealers have potential added benefits because of their bioactivity. After adequate endodontic therapy, osseous healing is largely dependent on the differentiation and activity of osteoblasts. We hypothesized that EndoSequence BC Sealer (Brasseler, Savannah, GA) and ProRoot ES (Dentsply Tulsa Dental Specialties, Johnson City, TN) have superior biocompatibility and osteogenic potential compared with Roth (Roth International, Chicago, IL) and AH Plus (Dentsply DeTrey Gmbh, Konstanz, Germany) sealers. METHODS: A murine osteoblast precursor cell line (IDG-SW3) was exposed to a wide range of concentrations for each of the sealers for 7 days. The relative cell viability was determined by luminescence assay based on adenosine triphosphate quantification (CellTiter-Glo [Promega, Madison, Wisconsin]). The osteogenic potential was determined by fluorescence microscopy of DMP-1 expression, alizarin red staining, and real-time reverse transcription polymerase chain reaction with primers specific for known markers of osteogenesis such as DMP-1, ALP, and Phex. Data were analyzed with 2-way analysis of variance or 1-way analysis of variance with the Bonferroni post hoc test. RESULTS: Both bioceramic sealers have excellent biocompatibility even at high concentrations. Conversely, cell death was detected when Roth and AH Plus were used at concentrations 100× lower than the bioceramic groups. Importantly, both bioceramic sealers significantly enhanced osteoblastic differentiation although greater responses were noted with EndoSequence BC Sealer. This was evidenced by increased DMP-1 expression, robust up-regulation of osteogenic marker gene expression, and superior mineral deposition. Osteoblastic differentiation and function were significantly impaired when Roth or AH Plus sealer was used. CONCLUSIONS: EndoSequence BC Sealer and ProRoot ES were significantly more biocompatible and promoted osteoblastic differentiation, a bioactivity not found in AH Plus and Roth sealers.

Sisters in hereditary breast and ovarian cancer families: communal coping, social integration, and psychological well-being.

OBJECTIVE: We investigated the association between psychological distress and indices of social integration and communal coping among sisters from hereditary breast and ovarian cancer (HBOC) families. SAMPLE AND METHODS: Sixty-five sisters from 31 HBOC families completed the Brief Symptom Inventory-18 and the Colored Eco-Genetic Relationship Map, which identified members of participants' social support networks. Hierarchical linear models were used for all analyses to account for the clustering of sisters within families. RESULTS: Intra-family correlation coefficients suggested that sisters shared perceptions of breast cancer risk and worry, but not ovarian cancer risk and worry. Further, sisters demonstrated shared levels of anxiety and somatization, but not depressive symptoms. Communal coping indices quantifying shared support resources were negatively related to anxiety and somatization. The number of persons with whom cancer risk information was shared exhibited a positive trend with somatization. Social integration, as measured by the size of participants' emotional support network, was negatively associated with anxiety. Lower depression scores were observed among participants with more persons playing multiple support roles and fewer persons providing tangible assistance. CONCLUSION: Understanding how support relationships impact well-being among persons adjusting to HBOC risk, and the particular role of family in that process, will facilitate developing appropriate management approaches to help cancer-prone families adjust to their cancer risk.