This week we introduce Lindsay Middelton R.N., C.G.C, who is a genetic counsellor with the National Cancer Institute (NCI) of the National Institutes of Health, USA.

Lindsay is part of the Urologic Oncology Branch at the NCI, which treats patients affected with inherited and sporadic forms of genitourinary cancer, and performs clinical trials as well as basic research. Teams at the NCI, in collaboration with researchers worldwide, identified the genes for BHD (Nickerson et al., 2002), Von-Hippel Lindau disease (Latif et al., 1993), and hereditary papillary renal cell cancer (Schmidt et al., 1997); work at the NCI on BHD has been ongoing since the 1990s. The NCI is in fact currently sponsoring a study investigating the genetics of BHD, “Genetic Analysis of Birt Hogg-Dube Syndrome and Characterization of Predisposition to Kidney Cancer”, which is recruiting participants.

As a genetic counsellor, Lindsay meets with individuals diagnosed with a hereditary condition. Genetic counsellors are often a very significant source of resources and support to families, offering guidance on understanding genetics, advice on managing a genetic condition, and time to discuss concerns, such as informing family members about a positive diagnosis. More information on genetic counselling and links for finding a local counsellor are available here.

With over 15 years’ experience in BHD and having met hundreds of patients, Lindsay’s specialist experience and knowledge is invaluable. As a member of the NCI team dedicated to undertaking translational research in kidney cancer disorders, Lindsay has travelled the United States to undertake clinical research into BHD and is an author on a number of papers. Lindsay Middelton’s research interests also include analysing nursing developments in cancer genetics as well as inherited kidney disorders.

The BHD community has also been fortunate to benefit from Lindsay’s expertise at the BHD Symposium, where she has been graciously leading the Patient and Family sessions (see summaries from the Third BHD Symposium and the Fourth BHD Symposium).

For a closer insight into Lindsay’s work, have a look at her Video Interview. There is also an audio-only version available, as well as a written transcript. The following publications also give an overview of Lindsay Middelton’s research:

• Middelton L, Dimond E, Calzone K, Davis J, & Jenkins J (2002). The role of the nurse in cancer genetics. Cancer nursing, 25 (3), 196-206 PMID: 12040228
• Singer EA, Bratslavsky G, Middelton L, Srinivasan R, & Linehan WM (2011). Impact of genetics on the diagnosis and treatment of renal cancer. Current urology reports, 12 (1), 47-55 PMID: 21128028
• Zbar B, Glenn G, Merino M, Middelton L, Peterson J, Toro J, Coleman J, Pinto P, Schmidt LS, Choyke P, & Linehan WM (2007). Familial renal carcinoma: clinical evaluation, clinical subtypes and risk of renal carcinoma development. The Journal of urology, 177 (2) PMID: 17222609

www.bhdsyndrome.org – the primary online resource for anyone interested in BHD Syndrome.

Mutations in genes such as FLCN and VHL are implicated in the development of renal cell carcinoma (RCC). However, DNA methylation and transcriptional silencing at gene promoters can also be involved, as is the case with RASSF1A, which is rarely mutated but often inactivated by promoter methylation in sporadic RCC (see Morris & Maher, 2010 for a review). A study by Ricketts et al. (2012) has now taken this concept further by revealing novel genes that also appear to be methylated in sporadic kidney cancer.

The authors used the Illumina Infinium HumanMethylation27 BeadChip array to directly assay the DNA methylation status of >14,000 genes in 38 samples of sporadic RCC and 9 normal kidney samples. Aberrant hyper- and hypo-methylation was observed in several (but not all) tumours when compared to control samples. Closer analysis identified a number of genes that were both highly methylated and had a functional relevance to cancer. In addition, this technique was verified using bisulphite sequencing and combined bisulfite restriction analysis (CoBRA).

Cluster analysis was used to group these genes into 5 categories depending on their level of methylation, and no association was observed between these groups and VHL mutation status. 6 genes (SLC34A2, OVOL1, DLEC1, TMPRSS2, SST and BMP4) were then investigated further and were shown to be methylated in a variety of RCC cell lines. These genes were also re-expressed in these cells after the addition of 5-Aza-2’-deoxycytidine, a DNA demethylating agent. Furthermore, the 6 genes exhibited both tumour-specific methylation and reduced expression when compared to normal kidney controls.

To investigate the functional impact of these genes, siRNA was used to knockdown OVOL1, SST and DLEC1 in HEK293 cells. Upon knockdown, anchorage-independent growth was significantly increased, especially with the OVOL1 knockdown. Moreover, real-time RT-PCR demonstrated that OVOL1 knockdown led to an increase in the expression of the proto-oncogene MYC (when compared to siRNA controls). OVOL1 is a zinc-finger transcription factor that is a downstream target of TGF-β signalling, and it has not been previously associated with human cancer.

In summary, this study has implicated a number of genes in the development of sporadic RCC, which could be used to develop novel biomarkers and (epigenetic) therapies. Khoo et al. (2003) noted that FLCN could also be inactivated by DNA methylation in sporadic RCC. Consequently, could DNA methylation play a role in BHD syndrome, for example, where no known FLCN mutations have been identified?

• Khoo SK, Kahnoski K, Sugimura J, Petillo D, Chen J, Shockley K, Ludlow J, Knapp R, Giraud S, Richard S, Nordenskjöld M, Teh BT. Inactivation of BHD in sporadic renal tumors. Cancer Res. 2003 Aug 1;63(15):4583-7. PMID: 12907635
• Morris MR, Maher ER. Epigenetics of renal cell carcinoma: the path towards new diagnostics and therapeutics. Genome Med. 2010 Sep 3;2(9):59. PMID: 20815920
• Ricketts CJ, Morris MR, Gentle D, Brown M, Wake N, Woodward ER, Clarke N, Latif F, Maher ER. Genome-wide CpG island methylation analysis implicates novel genes in the pathogenesis of renal cell carcinoma. Epigenetics. 2012 Mar;7(3):278-90. PMID: 22430804

www.bhdsyndrome.org – the primary online resource for anyone interested in BHD Syndrome.

Around 30 patients and family members participated in the Fourth BHD Symposium in Cincinnati, Ohio last month. Such a record turnout had a significant effect on the flow of the Symposium – for example, by providing researchers with many opportunities to meet and engage with families – and is evidence of an active BHD community. As readers of this blog will be aware, people affected by BHD and other rare conditions are widely spread across the globe; we hope this meeting will be an encouraging catalyst for future events as well as patient-led initiatives.

Following the welcoming reception on the 28th, the core patient and family activities took place on the 29th. After the clinical presentations, including an appraisal of screening guidelines for kidney tumours, (of which more details can be found in the previous Symposium live update post and Highlights post), a genetic counsellor from the NIH led a full afternoon of sharing experiences, reviewing research, and brainstorming new directions. We were also fortunate to welcome representatives from The LAM Foundation and the VHL Family Alliance to share strategies. LAM and VHL are conditions which have biological overlaps with BHD syndrome.

The Q&A portion of the patient sessions covered a number of concerns. For skin symptoms, the consensus is that there are a variety of treatments possible, but unfortunately these are not permanent and there are too few reports to evaluate which is the best temporary treatment. There is therefore need for studies into the best available treatment as well as investigations into experimental treatments. This year’s sessions also featured a greater emphasis on BHD lung symptoms, acting on observations from the Third BHD Symposium. Dr Frank McCormack, a pulmonologist from the University of Cincinnati responded to questions focusing on precautions, treatments and current understanding. Major points included the reiteration that BHD lung symptoms are not considered to be progressive and that air travel does not seem to pose a significant risk for pneumothorax, as investigated by Taveira-DaSilva et al., 2009. However, each person is different, so it may be helpful to speak with a doctor individually about air travel. Flu immunisations and smoking cessation were also recommended.

Additionally, following on from suggestions, BHDSyndrome.org will soon feature a private Forum for patients. There is also a private messaging system already in place, where any registered user can message someone who has posted to the Forum and opted into the system. If you have any suggestions or comments for BHDSyndrome.org, please complete the feedback form, which is always available on the For Families menu, the For Researchers menu and the Contact Us page.

All comments on the Symposium were greatly appreciated. We are excited to begin planning for the Fifth BHD Symposium and implementing ideas for improving the programme. Thank you for participating in the Fourth BHD Symposium!

• Taveira-DaSilva, A., Burstein, D., Hathaway, O., Fontana, J., Gochuico, B., Avila, N., & Moss, J. (2009). Pneumothorax After Air Travel in Lymphangioleiomyomatosis, Idiopathic Pulmonary Fibrosis, and Sarcoidosis. Chest, 136 (3), 665-670.

www.bhdsyndrome.org – the primary online resource for anyone interested in BHD Syndrome.

At the end of March, more than 90 BHD researchers, clinicians and patients attended the Fourth BHD Symposium, which was held in Cincinnati, Ohio, USA. The Symposium was the largest BHD meeting to date and it was a huge success. Talks were held in the impressive Vontz Center at the University of Cincinnati, which was an excellent location for the meeting. Representatives from the VHL Family Alliance, the LAM Foundation and the TS Alliance were also present, bringing together related rare disease organisations.

Highlights included invited talks from Dr Marston Linehan of the National Cancer Institute at the NIH, who spoke about the treatment of BHD kidney cancer and recommendations for screening, and Professor Kuniaki Seyama of Juntendo University, Japan, who discussed pulmonary manifestations of BHD. Other invited talks were given by Professor George Thomas of the University of Cincinnati, Professor Arnim Pause of McGill University and Professor Dr Maurice van Steensel of the University Hospital Maastricht.

Several clinicians took part in an open panel discussion, where screening recommendations and surveillance of BHD patients was further discussed. All clinicians agreed that kidney screening should begin around 21 years of age, but the frequency of MRI surveillance varied in different countries from every 1- 3 years.

Another interesting talk was given by Dr Fred Menko of the VU Medical Center in Amsterdam, who described FMDF, a disease with similar skin symptoms to BHD. A paper describing FMDF, summarised in a previous blog post, found that FLCN mutations were not associated with this disease. It is hoped that FMDF can provide insight into the skin symptoms of BHD.

Basic science highlights included an update on the gene therapy work given by Dr Suet-Ping Wong of Imperial College London, and the description of a zebrafish model of BHD presented by Dr Emma Kenyon of the University of Birmingham, UK. Scientists took part in an interactive session where experience and advice was shared regarding lab experiments and reagents. Out of this discussion, we are now planning an update to our Resources Forum.

The Symposium also included a dinner at the Verdin Bell Event Center, a poster session and a separate patient and family meeting. Highlights of this patient meeting will be available soon in a separate blog post. Finally, we would like to say a big thank you to all those who participated in the Symposium and contributed to its success, in particular Professor Maria Czyzyk-Krzeska who did an excellent job as the chair of the Scientific Organising Committee. We hope to see you all again next year!

www.bhdsyndrome.org – the primary online resource for anyone interested in BHD Syndrome.

At the beginning of March, Amsterdam hosted the 10^th International Congress on Targeted Anticancer Therapies, organised by the NDDO Education Foundation. The congress discussed all aspects of targeted therapy, from the identification of targets and the design of drugs, to clinical trials and the observed benefits for patients.

Dr Elizabeth Eisenhauer of Queen’s University, Canada presented the NDDO Honorary Award lecture, summarising the state of targeted therapy today. Targeted therapy has been in development for the past 15 years, and since 1999 there have been almost 15,000 Phase I, II and III clinical trials using targeted agents. Despite this large amount of research, there are still several problems associated with targeted treatments. These include choosing the right target, selecting the right patients to receive the treatment, side effects from the drugs and also drug resistance.

This year’s meeting included a session on epigenetic targets, a class of genes that are frequently found to be mutated in cancers. In renal cell carcinoma, several epigenetic regulators such as PBRM1, SETD2 and BAP1 have been found to be mutated. The identification of these genes has been discussed in previous blogs (here, here and here respectively). Dr Susan Bates from the National Cancer Institute at the NIH discussed the variety of epigenetic regulators that could be targeted in cancer therapy. Although mutations in epigenetic genes have been identified, it is unclear whether these mutations are the initiating event in tumourigenesis, or whether they are involved in maintaining the phenotype.

Sunitinib, a receptor tyrosine kinase inhibitor, is a targeted therapy approved by the FDA for the treatment of RCC. The drug works by inhibiting the formation of new blood vessels (known as angiogenesis), which effectively starves and destroys the tumour. Although Sunitinib has shown promising effects, drug resistance is a common problem. Prof. dr Henk Verheul, from the VU Medical Center in Amsterdam, discussed this resistance and suggested that it may be caused by sequestration of Sunitinib in lysosomes.

The TAT congress was a great overview of targeted therapy today. Although progress is being made, there are still many hurdles to be overcome in the development of targeted therapies. Finally, keep an eye on the Conference and Events page for details of upcoming meetings which are of interest to those involved in BHD syndrome. Over the next few weeks we will also be bringing you conference reports from the Fourth BHD Symposium, which was held recently in Cincinnati, USA.

www.bhdsyndrome.org – the primary online resource for anyone interested in BHD Syndrome.

A blog post from January last year described a paper by Purdue et al. (2011) in which a genome-wide association study (GWAS) identified two renal cell carcinoma (RCC) susceptibility loci within HIF2α (on chromosome 2) and an uncharacterised intergenic region (on chromosome 11). A recent study using VHL-deficient RCC cells has taken this work further by showing that this intergenic region contains a number of HIF2α and HIF1β binding sites within a previously unknown enhancer of cyclin D1 expression (Schödel et al., 2012).

Using chromatin immunoprecipitation (ChIP) with high-throughput sequencing, Schödel et al. (2012) detected HIF2α and HIF1β binding sites on chromosome 11 in the VHL-defective 786-O RCC cell line (but not in the pVHL-positive MCF-7 breast cancer cell line). ChIP with quantitative PCR (ChIP-qPCR) confirmed this binding in human RCC tissue, and hypoxia responsive elements (HREs) were also identified in this region.

This intergenic RCC susceptibility locus lacks the characteristics of a gene promoter. Consequently, a number of experiments were conducted to see if it was a transcriptional enhancer. Formaldehyde-assisted isolation of regulatory elements showed that this region had reduced nucleosome occupancy in 786-O cells (but not in MCF-7 cells). Similarly, ChIP-qPCR demonstrated that this locus had a variety of epigenetic marks that are characteristic of active enhancers (and not promoters) exclusively in the 786-O cells. Additionally, ChIP-qPCR in these cells showed that this locus had low levels of RNA polymerase II, which catalyses DNA transcription at promoters. Lastly, expression of a reporter gene was also enhanced by these sequences, and this effect was abrogated when the HREs were mutated. In total, these results suggest that the chromosome 11 RCC susceptibility locus contains a HIF-dependent transcriptional enhancer.

Further experiments showed that HIF binding and DNA accessibility was absent from a variety of cancerous and non-cancerous cell lines which expressed wild-type VHL, but was present in all VHL-deficient cell lines tested. In addition, genome-wide expression analysis in renal tumours and 786-O cells identified the nearby CCND1 gene (which encodes cyclin D1) as a HIF-regulated gene on chromosome 11. High-resolution fluorescent in situ hybridisation and chromosome conformation capture demonstrated that the HIF-binding enhancer and the CCND1 promoter physically associated in 786-O cells (but not in MCF-7 cells). These findings suggest that the chromosome 11 susceptibility locus is a cell type-specific long-range enhancer of the cyclin D1 gene.

The authors also saw that different polymorphisms in this region affect DNA accessibility and the binding of HIF2α, HIF1β and RNA polymerase II. Accordingly, these sequence variations could lead to the differential expression of cyclin D1, which offers a potential mechanism for the susceptibility effects observed in this region.

Given the connection between HIF signalling and BHD syndrome (Preston et al., 2010), a GWAS could uncover similar RCC susceptibility loci that are specific to FLCN-deficient cells. In particular, the potential links between FLCN, the HIF pathway and cell cycle control could help us to further understand the tumourigenic phenotypes observed in BHD syndrome.

• Preston RS, Philp A, Claessens T, Gijezen L, Dydensborg AB, Dunlop EA, Harper KT, Brinkhuizen T, Menko FH, Davies DM, Land SC, Pause A, Baar K, van Steensel MA, & Tee AR (2011). Absence of the Birt-Hogg-Dubé gene product is associated with increased hypoxia-inducible factor transcriptional activity and a loss of metabolic flexibility. Oncogene, 30 (10), 1159-73 PMID: 21057536
• Purdue MP, Johansson M, Zelenika D, Toro JR, Scelo G, Moore LE, Prokhortchouk E, Wu X, Kiemeney LA, Gaborieau V, Jacobs KB, Chow WH, Zaridze D, Matveev V, Lubinski J, Trubicka J, Szeszenia-Dabrowska N, Lissowska J, Rudnai P, Fabianova E, Bucur A, Bencko V, Foretova L, Janout V, Boffetta P, Colt JS, Davis FG, Schwartz KL, Banks RE, Selby PJ, Harnden P, Berg CD, Hsing AW, Grubb RL 3rd, Boeing H, Vineis P, Clavel-Chapelon F, Palli D, Tumino R, Krogh V, Panico S, Duell EJ, Quirós JR, Sanchez MJ, Navarro C, Ardanaz E, Dorronsoro M, Khaw KT, Allen NE, Bueno-de-Mesquita HB, Peeters PH, Trichopoulos D, Linseisen J, Ljungberg B, Overvad K, Tjønneland A, Romieu I, Riboli E, Mukeria A, Shangina O, Stevens VL, Thun MJ, Diver WR, Gapstur SM, Pharoah PD, Easton DF, Albanes D, Weinstein SJ, Virtamo J, Vatten L, Hveem K, Njølstad I, Tell GS, Stoltenberg C, Kumar R, Koppova K, Cussenot O, Benhamou S, Oosterwijk E, Vermeulen SH, Aben KK, van der Marel SL, Ye Y, Wood CG, Pu X, Mazur AM, Boulygina ES, Chekanov NN, Foglio M, Lechner D, Gut I, Heath S, Blanche H, Hutchinson A, Thomas G, Wang Z, Yeager M, Fraumeni JF Jr, Skryabin KG, McKay JD, Rothman N, Chanock SJ, Lathrop M, & Brennan P (2011). Genome-wide association study of renal cell carcinoma identifies two susceptibility loci on 2p21 and 11q13.3.Nature genetics, 43 (1), 60-5 PMID: 21131975
• Schödel J, Bardella C, Sciesielski LK, Brown JM, Pugh CW, Buckle V, Tomlinson IP, Ratcliffe PJ, & Mole DR (2012). Common genetic variants at the 11q13.3 renal cancer susceptibility locus influence binding of HIF to an enhancer of cyclin D1 expression. Nature genetics, 44 (4), 420-5 PMID: 22406644

www.bhdsyndrome.org – the primary online resource for anyone interested in BHD Syndrome.

We are here live at the Fourth BHD Symposium, at the University of Cincinnati. The University of Cincinnati has not only been called one of the world’s most beautiful campuses (by Forbes magazine), but also has an illustrious history in medical research, including the development of the first antihistamine and the first oral polio vaccine.

The Symposium began on Wednesday evening, when attendees met for a reception in the Kingsgate Conference Centre. The reception gave everyone the chance to meet and catch up before the scientific and family sessions kicked off on Thursday.

After an introduction by the chair of the scientific organising committee, Professor Maria Czyzyk-Krzeska, the Thursday morning session began with an overview of the treatment and management of BHD kidney cancer, given by Dr W. Marston Linehan of the National Cancer Institute at the NIH. The rest of the morning focused on clinical talks given by presenters from around the world. Dr Koga from the Chiba University School of Medicine in Japan spoke about pulmonary cysts in 14 BHD families, and Dr Fred Menko of the VU Medical Center in Amsterdam spoke about familial multiple discoid fibromas (FMDF), a novel syndrome with similar skin symptoms to BHD.

The morning session also included an interesting and dynamic panel discussion, led by researchers representing different health systems and approaches. Topics such as the age at which to begin kidney screening and the management of lung symptoms were discussed.

After the panel discussion, the scientific sessions focused on more basic science, with invited talks from Professor George Thomas of the University of Cincinnati and Professor Arnim Pause of McGill University, Montreal. The families attended a separate session, led by Lindsay Middleton, a genetic counsellor at the NIH.

All attendees came together at the end of the day for the poster session, followed by a dinner at the Verdin Bell Event Centre. The Centre was originally St. Paul’s church, built in 1850 and decommissioned in 1975. Bought in 1981 by The Verdin Company, the oldest bell and clock company in North America, it has now been carefully restored and contains a wonderful collection of historic bells and clocks.

Many more exciting talks are still to come on the final day of the Symposium. Remember to watch out for our full reports from the Symposium over the next few weeks, covering the scientific and family sessions. Don’t forget to keep following the live Symposium news, as well as all other BHD syndrome news on Twitter, hashtag #BHDSyndrome.

www.bhdsyndrome.org – the primary online resource for anyone interested in BHD Syndrome.

The Fourth BHD Symposium is less than a week away, and this year it will be held in Cincinnati, USA, on 28^th to 30^th March. Registration is still open and more information regarding the entire Symposium can be found here.

In particular, the Fourth BHD Symposium agenda and Patient and family sessions schedule are now available. There will also be a social gathering to welcome everyone to the Symposium on Wednesday 28^th March from 6 pm at the Kingsgate Marriott.

From Thursday 29^th March, there will be a wide variety of clinical and basic science talks, including those from Dr Marston Linehan (National Cancer Institute, USA) and Professor Angela Christiano (Columbia University Medical Center, USA). As with previous years, there will be a poster session introducing yet more BHD research. In addition, patients and family members will be able to meet each other and experts in the BHD field during the Patient and family sessions.

Please do follow the meeting on our Twitter feed, hashtag #BHDSyndrome. Next week’s blog post will also be a live report from the Symposium, and there will be further highlights from Cincinnati in the weeks to come. We hope to see you there!

www.bhdsyndrome.org – the primary online resource for anyone interested in BHD Syndrome.

Recent advances in DNA sequencing have led to the discovery of genes and mutations which drive tumourigenesis. In this blog, we have previously described papers by Varela et al. (2011), Dalgliesh et al. (2011) and Guo et al. (2011) which all used exome sequencing to identify genes involved in kidney cancer (described here, here and here respectively). It is hoped that by understanding more about the cancer genome, effective targeted therapies can be developed.

A recent study by Xu et al. (2012) performed sequencing analysis on one ccRCC kidney tumour sample, this time by using single-cell exome sequencing. Exome sequencing was first used on a sample of ccRCC tissue to analyse the mutational status of VHL and PBRM1, the main drivers of ccRCC tumourigenesis. Three low frequency mutations in PBRM1 were identified, but no mutations were found in VHL.

Single-cell exome sequencing was then performed on 20 cells from the tumour sample and 5 cells from noncancerous adjacent tissue, which were used as controls. In total, 260 somatic mutations were identified in the cancer cells, compared with 12 somatic mutations in the healthy control cells. Mutations in 94 genes were identified.

The mutation pattern for each individual cell was compared and the results were found to be significantly different from each other. The authors therefore suggest that the tumour does not contain subpopulations of cells. Additionally, more than 70% of the mutations were classed as cell-specific, meaning they only occurred in a small fraction of the cells. Both of these results demonstrate the high complexity and heterogeneity of this tumour sample.

The results from this study were compared to the recent sequencing study by Guo et al. (2011). Four of the 94 genes identified here were also mutated in the cohort of 98 patients studied by Guo et al. One gene of particular interest is AHNAK, which was mutated in 5 of the 99 patients from both studies. AHNAK activates protein kinase C and it is also involved in chromatin remodelling, a process in which related genes appear to be frequently mutated in RCC. Furthermore, protein-protein prediction methods used in this study suggest an interaction between AHNAK and HIF1A.

With regards to BHD syndrome, no FLCN mutations were identified in the RCC cells studied here. However, it would be interesting to perform a similar study using cells from a BHD tumour to identify other mutations which could contribute to tumourigenesis in BHD syndrome. The authors of this study suggest that ccRCC is more genetically complex than previously thought. Perhaps the same is true for BHD-related tumours and that there are other, as yet, unidentified mutations which promote tumourigenesis.

• Xu X, Hou Y, Yin X, Bao L, Tang A, Song L, Li F, Tsang S, Wu K, Wu H, He W, Zeng L, Xing M, Wu R, Jiang H, Liu X, Cao D, Guo G, Hu X, Gui Y, Li Z, Xie W, Sun X, Shi M, Cai Z, Wang B, Zhong M, Li J, Lu Z, Gu N, Zhang X, Goodman L, Bolund L, Wang J, Yang H, Kristiansen K, Dean M, Li Y, Wang J. Single-cell exome sequencing reveals single-nucleotide mutation characteristics of a kidney tumor. Cell. 2012 Mar 2;148(5):886-95. PMID: 22385958
• Varela I, Tarpey P, Raine K, Huang D, Ong CK, Stephens P, Davies H, Jones D, Lin ML, Teague J, Bignell G, Butler A, Cho J, Dalgliesh GL, Galappaththige D, Greenman C, Hardy C, Jia M, Latimer C, Lau KW, Marshall J, McLaren S, Menzies A, Mudie L, Stebbings L, Largaespada DA, Wessels LF, Richard S, Kahnoski RJ, Anema J, Tuveson DA, Perez-Mancera PA, Mustonen V, Fischer A, Adams DJ, Rust A, Chan-on W, Subimerb C, Dykema K, Furge K, Campbell PJ, Teh BT, Stratton MR, Futreal PA. Exome sequencing identifies frequent mutation of the SWI/SNF complex gene PBRM1 in renal carcinoma. Nature. 2011 Jan 27;469(7331):539-42. PMID: 21248752
• Dalgliesh GL, Furge K, Greenman C, Chen L, Bignell G, Butler A, Davies H, Edkins S, Hardy C, Latimer C, Teague J, Andrews J, Barthorpe S, Beare D, Buck G, Campbell PJ, Forbes S, Jia M, Jones D, Knott H, Kok CY, Lau KW, Leroy C, Lin ML, McBride DJ, Maddison M, Maguire S, McLay K, Menzies A, Mironenko T, Mulderrig L, Mudie L, O’Meara S, Pleasance E, Rajasingham A, Shepherd R, Smith R, Stebbings L, Stephens P, Tang G, Tarpey PS, Turrell K, Dykema KJ, Khoo SK, Petillo D, Wondergem B, Anema J, Kahnoski RJ, Teh BT, Stratton MR, Futreal PA. Systematic sequencing of renal carcinoma reveals inactivation of histone modifying genes. Nature. 2010 Jan 21;463(7279):360-3. PMID: 20054297
• Guo G, Gui Y, Gao S, Tang A, Hu X, Huang Y, Jia W, Li Z, He M, Sun L, Song P, Sun X, Zhao X, Yang S, Liang C, Wan S, Zhou F, Chen C, Zhu J, Li X, Jian M, Zhou L, Ye R, Huang P, Chen J, Jiang T, Liu X, Wang Y, Zou J, Jiang Z, Wu R, Wu S, Fan F, Zhang Z, Liu L, Yang R, Liu X, Wu H, Yin W, Zhao X, Liu Y, Peng H, Jiang B, Feng Q, Li C, Xie J, Lu J, Kristiansen K, Li Y, Zhang X, Li S, Wang J, Yang H, Cai Z, Wang J. Frequent mutations of genes encoding ubiquitin-mediated proteolysis pathway components in clear cell renal cell carcinoma. Nat Genet. 2011 Dec 4;44(1):17-9. PMID: 22138691

www.bhdsyndrome.org – the primary online resource for anyone interested in BHD Syndrome.

Our lab-profile for this month introduces the work of Professor Arnim Pause, an Associate Professor in the Department of Biochemistry and a Canada Research Chair in Molecular Oncology at McGill University, Canada.

Much of Professor Pause’s early work involved researching eukaryotic translation initiation factors, such as eIF-4A and eIF-4E, which play an important role in protein biosynthesis (Dostie et al., 2000). Concurrent work focussed on the function of VHL, which was discovered to be involved in the ubiquitination of proteins and their degradation by the proteasome (Iwai et al., 1999).

Currently, the Pause lab continues to characterise the function of the VHL protein (Kurban et al., 2008), as well as investigating the role of a His domain-containing protein tyrosine phosphatase encoded by the tumour suppressor gene candidate PTPN23 (Gingras et al., 2009). Professor Pause also studies the role of several proteins in the life cycle of the Hepatitis C virus (Tedbury et al., 2011).

Another major research interest of the Pause lab is the functional characterisation of the FLCN protein. Using mouse models and human cell lines, Hudon et al. (2010) noted that a loss of FLCN expression resulted in both elevated and reduced levels of mTOR signalling, which may account for the differing results observed in previous studies (see here for more information). The Pause lab also collaborated with Dr Andrew Tee’s group at Cardiff University to show that FLCN is associated with HIF signalling (Preston et al., 2011), which has been discussed in more detail in a previous blog post and lab-profile. Further studies have also been planned that make use of a transparent nematode called C. elegans. This multicellular model organism facilitates systematic molecular and genetic approaches, while still maintaining relevance to other eukaryotes such as humans.

To hear more about the research of the Pause lab, and why rare disease research is particularly important for medical research in general, please do watch our video interview (with its accompanying transcript and audio file). Alternatively, join us at the 4^th BHD Symposium on 28^th – 30^th March in Cincinnati, USA. Registration is still open and more information about the patient and family sessions can be found here.

• Dostie J, Ferraiuolo M, Pause A, Adam SA, Sonenberg N. A novel shuttling protein, 4E-T, mediates the nuclear import of the mRNA 5′ cap-binding protein, eIF4E. EMBO J. 2000 Jun 15;19(12):3142-56. PMID: 10856257
• Gingras MC, Zhang YL, Kharitidi D, Barr AJ, Knapp S, Tremblay ML, Pause A. HD-PTP is a catalytically inactive tyrosine phosphatase due to a conserved divergence in its phosphatase domain. PLoS One. 2009;4(4):e5105. Epub 2009 Apr 2. PMID: 19340315
• Hudon V, Sabourin S, Dydensborg AB, Kottis V, Ghazi A, Paquet M, Crosby K, Pomerleau V, Uetani N, Pause A. Renal tumour suppressor function of the Birt-Hogg-Dubé syndrome gene product folliculin. J Med Genet. 2010 Mar;47(3):182-9. Epub 2009 Oct 19. PMID: 19843504
• Iwai K, Yamanaka K, Kamura T, Minato N, Conaway RC, Conaway JW, Klausner RD, Pause A. Identification of the von Hippel-lindau tumor-suppressor protein as part of an active E3 ubiquitin ligase complex. Proc Natl Acad Sci U S A. 1999 Oct 26;96(22):12436-41. PMID: 10535940
• Kurban G, Duplan E, Ramlal N, Hudon V, Sado Y, Ninomiya Y, Pause A. Collagen matrix assembly is driven by the interaction of von Hippel-Lindau tumor suppressor protein with hydroxylated collagen IV alpha 2. Oncogene. 2008 Feb 7;27(7):1004-12. Epub 2007 Aug 13. PMID: 17700531
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www.bhdsyndrome.org – the primary online resource for anyone interested in BHD Syndrome.