Tjinta Brinkhuizen

Impact of ankylosing spondylitis on sick leave, presenteeism and unpaid productivity, and estimation of the societal cost.

Aim To describe the influence of ankylosing apondylitis (AS) on sick leave, presenteeism and unpaid work restrictions and to estimate related productivity costs. Methods 142 consecutive and unselected patients with AS under the care of rheumatologists participated in a longitudinal observational study and completed the Health and Labour Questionnaire (HLQ) assessing disease-related sick leave, presenteeism and restrictions in unpaid work over the previous 2 weeks. Logistic regressions explored which explanatory variables were associated with work outcome. Productivity loss was valued in monetary terms. Results Among 72 patients in paid employment, 12% had sick leave over a period of 2 weeks and 53% experienced an adverse influence of AS on work productivity while at work. Over this period they reported on average of 5.8 h sick leave and 2.4 inefficient working hours, for which they estimated an extra 1.9 h were needed to complete unfinished work. Among all patients (n=137), 71% had experienced restrictions in unpaid work during the previous 2 weeks with 42% needing help for these tasks for an average of 8 h. The annual production costs for the total group were €1451 (95% CI 425 to 2742) per patient for sick leave, €967 (95% CI 503 to 1496) to compensate for hours worked inefficiently while at work and €1930 (95% CI 1404 to 2471) to substitute loss of unpaid work production. Conclusion Patients with AS not only have substantial sick leave but also experience restrictions while being at work and when performing unpaid tasks. Limitations in physical functioning are strongly associated with work restrictions. Societal costs of formal and informal care are comparable with the costs of sick leave and presenteeism combined.

Absence of the Birt-Hogg-Dubé gene product is associated with increased hypoxia-inducible factor transcriptional activity and a loss of metabolic flexibility

Under conditions of reduced tissue oxygenation, hypoxia-inducible factor (HIF) controls many processes, including angiogenesis and cellular metabolism, and also influences cell proliferation and survival decisions. HIF is centrally involved in tumour growth in inherited diseases that give rise to renal cell carcinoma (RCC), such as Von Hippel–Lindau syndrome and tuberous sclerosis complex. In this study, we examined whether HIF is involved in tumour formation of RCC in Birt–Hogg–Dubé syndrome. For this, we analysed a Birt–Hogg–Dubé patient-derived renal tumour cell line (UOK257) that is devoid of the Birt–Hogg–Dubé protein (BHD) and observed high levels of HIF activity. Knockdown of BHD expression also caused a threefold activation of HIF, which was not as a consequence of more HIF1α or HIF2α protein. Transcription of HIF target genes VEGF, BNIP3 and CCND1 was also increased. We found nuclear localization of HIF1α and increased expression of VEGF, BNIP3 and GLUT1 in a chromophobe carcinoma from a Birt–Hogg–Dubé patient. Our data also reveal that UOK257 cells have high lactate dehydrogenase, pyruvate kinase and 3-hydroxyacyl-CoA dehydrogenase activity. We observed increased expression of pyruvate dehydrogenase kinase 1 (a HIF gene target), which in turn leads to increased phosphorylation and inhibition of pyruvate dehydrogenase. Together with increased protein levels of GLUT1, our data reveal that UOK257 cells favour glycolytic rather than lipid metabolism (a cancer phenomenon termed the ‘Warburg effect’). UOK257 cells also possessed a higher expression level of the L-lactate influx monocarboxylate transporter 1 and consequently utilized L-lactate as a metabolic fuel. As a result of their higher dependency on glycolysis, we were able to selectively inhibit the growth of these UOK257 cells by treatment with 2-deoxyglucose. This work suggests that targeting glycolytic metabolism may be used therapeutically to treat Birt–Hogg–Dubé-associated renal lesions.

Epigenetic Changes in Basal Cell Carcinoma Affect SHH and WNT Signaling Components

Background The genetic background of Basal Cell Carcinoma (BCC) has been studied extensively, while its epigenetic makeup has received comparatively little attention. Epigenetic alterations such as promoter hypermethylation silence tumor suppressor genes (TSG) in several malignancies. Objective We sought to analyze the promoter methylation status of ten putative (tumor suppressor) genes that are associated with Sonic Hedgehog (SHH), WNT signaling and (hair follicle) tumors in a large series of 112 BCC and 124 healthy control samples by methylation-specific PCR. Results Gene promoters of SHH (P = 0.016), adenomatous polyposis coli (APC) (P = 0.003), secreted frizzled-related protein 5 (SFRP5) (P = 0.004) and Ras association domain family 1A (RASSF1A) (P = 0.023) showed significantly more methylation in BCC versus normal skin. mRNA levels of these four genes were reduced for APC and SFRP5 in BCC (n = 6) vs normal skin (n = 6). Down regulation of SHH, APC and RASSF1A could be confirmed on protein level as well (P<0.001 for all genes) by immunohistochemical staining. Increased canonical WNT activity was visualized by β-catenin staining, showing nuclear β-catenin in only 28/101 (27.7%) of BCC. Absence of nuclear β-catenin in some samples may be due to high levels of membranous E-cadherin (in 94.1% of the samples). Conclusions We provide evidence that promoter hypermethylation of key players within the SHH and WNT pathways is frequent in BCC, consistent with their known constitutive activation in BCC. Epigenetic gene silencing putatively contributes to BCC tumorigenesis, indicating new venues for treatment.

Immunohistochemical Analysis of the Mechanistic Target of Rapamycin and Hypoxia Signalling Pathways in Basal Cell Carcinoma and Trichoepithelioma

Background Basal cell carcinoma (BCC) is the most common cancer in Caucasians. Trichoepithelioma (TE) is a benign neoplasm that strongly resembles BCC. Both are hair follicle (HF) tumours. HFs are hypoxic microenvironments, therefore we hypothesized that hypoxia-induced signalling pathways could be involved in BCC and TE as they are in other human malignancies. Hypoxia-inducible factor 1 (HIF1) and mechanistic/mammalian target of rapamycin (mTOR) are key players in these pathways. Objectives To determine whether HIF1/mTOR signalling is involved in BCC and TE. Methods We used immunohistochemical staining of formalin-fixed paraffin-embedded BCC (n = 45) and TE (n = 35) samples to assess activity of HIF1, mTORC1 and their most important target genes. The percentage positive tumour cells was assessed manually in a semi-quantitative manner and categorized (0%, <30%, 30–80% and >80%). Results Among 45 BCC and 35 TE examined, expression levels were respectively 81% and 57% (BNIP3), 73% and 75% (CAIX), 79% and 86% (GLUT1), 50% and 19% (HIF1α), 89% and 88% (pAKT), 55% and 61% (pS6), 15% and 25% (pMTOR), 44% and 63% (PHD2) and 44% and 49% (VEGF-A). CAIX, Glut1 and PHD2 expression levels were significantly higher in TE when only samples with at least 80% expression were included. Conclusions HIF and mTORC1 signalling seems active in both BCC and TE. There are no appreciable differences between the two with respect to pathway activity. At this moment immunohistochemical analyses of HIF, mTORC1 and their target genes does not provide a reliable diagnostic tool for the discrimination of BCC and TE.

Soft yellowish papules on the neck: a clinicopathological challenge

Soft yellowish papules on the neck: a clinicopathological challenge K. J. A. Frencken, M. N. K. Hacking, T. Brinkhuizen, M. A. Abdul Hamid and H. Martens Departments of Dermatology and Pathology, Maastricht University Medical Centre, Maastricht, the Netherlands; GROW, School for Oncology and Developmental Biology, Maastricht University, Maastricht, the Netherlands; and Department of Dermatology, Catharina Hospital Eindhoven, Eindhoven, the Netherlands

Locally advanced basal cell carcinoma has a distinct methylation and transcriptomic profile

flammatory hyperpigmentation HS scars. D. Comparison of a normal and mutant partial chromatograph of DNA from family UR-252. The insertion of 2-nt (CC) at nucleotide position 687 of human NCSTN is shown. E. The truncated protein is predicted to terminate at codon 261 after the addition of 31 novel amino acids within exon 6 of NCSTN. Figure S2. A. Family UR-253 with acne conglobata (AC) was previously reported by Chandramohan (S17). The pedigree consists of 23 members with seven affecteds. Squamous cell carcinoma in the perianal region was observed in one affected member. Affected individuals are shown with black symbols and normal individuals with clear symbols. Individuals with a slash across their symbol were deceased. B. Clinical manifestations of an affected subject (III-6) with AC lesions demonstrating inflammatory (papules, pustules, nodules and cysts) and non-inflammatory comedones: chest and abdomen, C. back. D. Partial nucleotide sequence from family UR-253 showing deletion of 2-nt (TG) at nucleotide 1799 in NCSTN, resulting in a stop codon (L600X). E. Comparison of translation products of the normal and mutated NCSTN allele. The truncated protein terminates at codon 600. Table S1. Details of primers for NCSTN, PSEN1, PSEN2, and PSENEN genes used in the present mutation analysis. Data S1. Materials and methods. Data S2. Supplementary References.

Abstract 4929: Hypermethylation of tumor suppressor gene promoters in basal cell carcinoma

Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC Background Basal-cell carcinoma (BCC) is the most frequent malignancy in Caucasians, with increasing incidence. Overactivity of the Sonic Hedgehog pathway is implicated in BCC etiology, however recent literature suggests that BCC carcinogenesis may be more complex. Silencing of tumor suppressor genes by promoter hypermethylation, influenced by hypoxia, results in deregulation of cancer-related pathways. Since BCC arises in a hypoxic micro-environment, we assumed that epigenetic modifications may play a major part in BCC tumorigenesis. In this pilot we investigated promoter hypermethylation of tumor suppressor genes selected for their possible involvement in BCC. Eventually, knowledge of epigenetic modifications in BCC may impact diagnosis, classification, and treatment. Methods DNA was isolated from 33 formalin fixed paraffin-embedded BCC of 3 different histological subtypes. Marked tumor tissue was macroscopically scraped from slides and collected. CpG island methylation status of 10 promoter regions was analysed by methylation-specific PCR on sodium bisulfite-treated DNA. Data were compared with 36 normal skin tissue samples. All data were analysed with SPSS version 17.0. We used the Pearson Chi-Square test. Statistical significance was defined as a p-value ≤0.05. Results DNA methylation levels of the examined candidate gene promoters in tumor tissue compared to normal skin tissue were significantly higher for the following genes: APC 100% vs. 47,1% (p<0,001); sFRP5 93,9% vs. 42,4% (p<0,001); SHH 90,9% vs. 50,0% (p< 0,001) and 81,8% vs. 56,3% (p= 0,029) for CylD. In addition, the PTCH1 promoter showed a tendency to hypermethylation. The remaining promoters of sFRP1, sFRP2, sFRP4, TSC1 and RASSF1A showed no hypermethylation. Conclusions From our preliminary data we conclude that in BCC, promoters of several tumor suppressor genes, which may normally inhibit BCC growth, show aberrant methylation resulting in silencing of these genes. As a consequence, signalling pathways implicated in cancerous growth such as the SHH-, WNT- and NFκB- routes may be affected. Our data provide evidence that epigenetic modifications may play a role in BCC tumorigenesis. Genome-wide analyses will be our next step to screen for more relevant genes and to chart the full extent of genomic methylation changes in BCC. This entirely new approach may lead to new insights into development and treatment of BCC. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4929.