Curriculum vitae of Dr. Esther Maina

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CURRICULUM VITAE

1. Family name: MAINA

2. First names: Esther Njoki Mwangi

3. Date of birth: 13th March 1978

4. Nationality: Kenyan

5. Civil status: Married

6. Education:

Institution: University of Birmingham (UK), Division of Paediatrics and Child HealthDate: 2000-04Degree/Diploma: Ph.D. (medical and molecular genetics)Institution: University of Essex (UK), School of Biological SciencesDate: 1997-2000Degree/Diploma: B.Sc. Hons (Biochemistry and Molecular Medicine), 2000

7. Language skills:

Language (1-5, 1 = excellent) Reading Speaking WritingEnglish 1 1 1Swahili 1 1 1Kikuyu (mother tongue) 1 1 1

8. Present position: Lecturer, Department of Biochemistry, Chiromo Campus, University of Nairobi (Kenya)

Years with the organisation: Since October 2009

9. University and departmental appointments

- Course Co-ordinator, Master of Surgery, College of Health Science

- Course Co-ordinator, BSc Medical laboratory science Level II, College of Health Science

- Member, Scientific committee, College of agriculture and veterinary science

- Member, Postgraduate committee, College of agriculture and veterinary science (Oct 2009- 2013)

- Member, Curriculum committee, College of agriculture and veterinary science

- Member, Time-tabling committee, College of agriculture and veterinary science (Oct 2009 – 2013)

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10. Membership of professional bodies:

- International Federation of University Women (IFUW)

- Third World Organization for Women in Science (TWOWS)

- African Women’s Forum on Science and Technology (AWFST)

- African Technology Policy Studies Network

- Biochemical Society of Kenya

- Honorary member of the Nairobi University Biochemistry Student Association

11. Academic/professional honours and awards

2012: Awarded the 2012 NCST PhD grant (KES 1.2 M) towards facilitation of PhD studies to qualified student

2010: Awarded a travel grant (KES 77,000) to attend an Epidemiology Course by the Regional Aids Training Network (RATN), October 2010, Nairobi

2010: Awarded an attendance grant for the Qiagen workshop, October 2010, Nairobi

2004-6: Awarded conference travel grants by the British Society of Human Genetics

2003: Awarded a conference travel grant by the American Association of Cancer Research, June 2003 Washington D.C, USA

2003: Awarded first prize in oral presentations at the Division of Paediatrics and Child Health Research Day, May 2003, University of Birmingham (UK)

2000: Awarded the Pfizer CASE PhD studentship, October 2000, Birmingham (UK)

1997: Awarded the Borough of Ealing, Local education authority grant for undergraduate studies

12. On-going Research work:

a. PhD Project supervision: “Development of an FT- NIR calibration models for estimating /detecting Aflatoxin B1 content in Maize kernels for further maize quality testing “ by Gladys Bosibori Bichanga, Department of Biochemistry

b. Master’s project supervision: “INVESTIGATION OF GENOTOXICITY OF SOLAR DISINFECTED WATER” by Anne Moraa Kerandi, Department of Chemistry. Collaborator: Prof. Sir Shem Wandiga

c. Master’s project supervision: “INVESTIGATION OF DISCORDANT HIV COUPLES” By Virginia Njau, Department of Biochemistry. Collaborator: Prof. Joseph Mukuria

d. Epidemiological study on Prader-willi syndrome (PWS) incidence in Kenya. Collaborator: Dr. Frank Njenga

e. Plants of Medicinal use Established for the 3rd year undergraduate students to mentor them in project inception and management, Department of Biochemistry

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13. Employment record:

Date: Since October 2009Location: Nairobi (Kenya)Company: University of Nairobi, Position: Lecturer, Department of Biochemistry, Chiromo CampusDescription: Teaching biochemistry, genetics and molecular biology to both undergraduate and

postgraduate students; actively involved in proposal writing for research grants; departmental representative on university Committees on Curriculum Development, Postgraduate Studies, and Timetabling; member of departmental committees on Strategic Planning and Higher Degrees.

Date: April-October 2009Location: Kampala (Uganda)Company: Kampala International UniversityPosition: Lecturer, Department of BiochemistryDescription: Teaching genetics and molecular biology to both undergraduate (MBCHB) and

postgraduate (master’s level) students; prepared curriculum for the course and actively involved in proposal writing for research grants.

Date: February-November 2008Location: Birmingham (UK)Company: University of BirminghamPosition: Post-Doctoral Research Fellow CRUK/LecturerDescription: Working to establish the role of the transcription factor Oct-1 in chronic lympho-

blastic lymphoma, and its effect on the integrity of the DNA repair machinery with respect to Ataxia Telangiectasia Mutated (ATM), a protein that is recruited to sites of DNA damage for repair purposes.

Date: 2006-08Location: Birmingham (UK)Company: University of BirminghamPosition: Post-Doctoral Research Fellow CRUK/LecturerDescription: Worked on the role of EBV in Hodgkin’s Lymphoma (HL), including work towards

establishing the relationship between EBV infection of HL precursor cells, Hodgkin-Reed Sternberg (HRS) cells, and the integrity of DNA repair.

Date: 2004-06Location: Birmingham and Cambridge (UK)Company: University of Birmingham and Cambridge UniversityPosition: Post-Doctoral Joint FellowshipDescription: Worked on Prader-Willi syndrome (PWS) , which is a complex human genetic

disease (imprinting disorder) arising from lack of expression of paternally inheritedand imprinted genes on chromosome 15q11-q13, including mutation analyses and gene-expression studies on putative PWS genes.

Date: 2003-04Location: Birmingham (UK)Company: University of BirminghamPosition: Post-Doctoral Research AssociateDescription: Carried out functional studies on novel, pVHL target genes, including siRNA, cloning,

cell culture, transfections, growth suppression and migration assays.Date: 2000-03 Location: Birmingham (UK)Company: Pfizer Genetic TechnologyPosition: Ph.D. industrial placement (part-time), Genetic Technology DepartmentDescription: Conducted experiments in a Good Laboratory Practice (GLP)-compliant research

laboratory; gained experience in teamwork, organisational and communication skills, problem-solving, and meeting tight deadlines.

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14. Key qualifications:

I am a practicing scientist in the Department of Biochemistry at the University of Nairobi. I am actively involved in teaching at the undergraduate and postgraduate level, I manage a programmes of laboratory research (including proposal writing), and play a role in academic management at the departmental and university level as a representative of the department in several faculty committees including time-tabling, postgraduate, curriculum and scientific committees in the Faculty of Veterinary Medicine .

As far as teaching is concerned, I have taught Biochemistry, Genetics and Molecular Biology to medical students (BMS and MBCHB degrees) and Master’s students in the biomedical sciences. I have also been involved in the supervision of Ph.D. students, including guiding them through their projects on a day-to-day basis in the laboratory and providing guidance and advice throughout their projects.

My research interests are in the field of genetics and molecular biology. I earned my Ph.D. with a dissertation on the use of microarray technology to elucidate the cell-signalling pathway in the Von Hippel- Lindau (VHL) tumour suppressor gene, which is a causative gene for the familial cancer syndrome. The objectives of my Ph.D. thesis were:

• To use microarray technology to identify novel, pVHL target genes and genes whose expression is affected by the different VHL mutations, thus causing the different pVHL phenotypes; and

• To carry out functional studies on putative, pVHL target genes.

Previously, for my undergraduate degree, I studied the in vitro effects of isothiocyanates (sulforaphane) on the growth and viability of human leukaemia (HL60) cell line.

I have excellent organisational and communication skills, both acquired during my Ph.D. studies and in the scientific and retail industries. I have a high level of motivational, leadership and transferable skills, gained through social activities and teaching.

15. Other skills:

Laboratory techniques, including: microarray analysis; real-time PCR, DNA and RNA extraction and manipulation, PCR, DNA quantification and sequencing; SNP and mutation detection; epigenetic techniques (Bisulphite modifications, 5-aza treatment); mammalian cell culture, primary cell culture, cloning and bacterial transformation; siRNA mammalian cell transfection; immunofluorescence, immunoprecipitation (CO-IP), Comet assay, FACS sorting, and protein expression analysis.

-Extensive knowledge of Microsoft Office programs, web-based bioinformatics packages, and various packages for microarray data analysis.-Mentorship: Mentoring young people and empowering young women to succeed in science and technology disciplines-Extensive knowledge of Microsoft Office programs, web-based bioinformatics packages, and various packages for microarray data analysis.

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16. Selected scientific publications

2012. Esther N. Maina , Tessa Webb , John R Arrand , Joyce Whittington , Sarita Soni, Harm Boer , David Clarke , Anthony J. Holland. Investigation of Prader-Willi-like Phenotype using a Whole Genome Array. African Journal of Pharmacology and Therapeutics Vol. 1 No. 3 Pages 84-91, 2012

2012. Maina E.N.M. Looking to the study of genes for cancer diagnoses and Treatment. (Invited mini-review) Your Doctor Magazine 2012 Jul; 4 (5-6)

2009. Bose S, Yap LF, Fung M, Starzcynski J, Saleh A, Morgan S, Dawson C, Chukwuma MB, Maina E, Buettner M, Wei W, Arrand J, Lim PV, Young LS, Teo SH, Stankovic T, Woodman CB, Murray PG. The ATM tumour suppressor gene is down-regulated in EBV-associated nasopharyngeal carcinoma. J Pathol. 2009 Feb; 217(3):345-52.

2009. Marston E, Weston V, Jesson J, Maina E, McConville C, Agathanggelou A, Skowronska A, Mapp K, Sameith K, Powell JE, Lawson S, Kearns P, Falciani F, Taylor M, Stankovic T. Stratification of pediatric ALL by in vitro cellular responses to DNA double-strand breaks provides insight into the molecular mechanisms underlying clinical response. Blood. 2009 Jan 1;113 (1):117-26.

2008. Tessa Webb, Esther N. Maina, Sarita Soni, Joyce Whittington, Harm Boer, David Clarke, Anthony Holland. In search of the psychosis gene in people with Prader-Willi syndrome. Am J Med Genet A. 2008 Apr 1; 146 (7):843-53.

2008. S. Soni, J. Whittington, A. J. Holland, T. Webb, E. N. Maina, H. Boer and D. Clarke. The phenomenology and diagnosis of psychiatric illness in people with Prader-Willi syndrome. Psychol. Med. 2008 Jan 4:1-10.

2007. Maina EN, Webb T, Soni S, Whittington J, Boer H, Clarke D, Holland A. Analysis of candidate imprinted genes in PWS subjects with atypical genetics: a possible inactivating mutation in the SNURF/SNRPN minimal promoter. J. Hum. Genet. 2007; 52(4):297-307.

2007. Soni S, Whittington J, Holland AJ, Webb T, Maina E, Boer H, Clarke D. The course and outcome of psychiatric illness in people with Prader-Willi syndrome: implications for management and treatment. J Intellect Disabil Res. 2007 Jan; 51(Pt 1):32-42.

2006. Maina EN, Abdulrahman M, Morris MR, Zatyka M, Raval RR, Banks RE, Wiesener MS, Richards FM, Johnson CM, Latif F, Maher ER. Identification of novel VHL targets that are associated with the development of renal cell carcinoma. Oncogene. 2007 Mar 8; 26(11):1661-72. Epub 2006 Sep 25.

2006. Smith UM, Consugar M, Tee LJ, McKee BM, Maina EN, Whelan S, Morgan NV, Goranson E, Gissen P, Lilliquist S, Aligianis IA, Ward CJ, Pasha S, Punyashthiti R, Malik Sharif S, Batman PA, Bennett CP, Woods CG, McKeown C, Bucourt M, Miller CA, Cox P, Algazali L, Trembath RC, Torres VE, Attie-Bitach T, Kelly DA, Maher ER, Gattone VH 2nd, Harris PC, Johnson CA. The transmembrane protein meckelin (MKS3) is mutated in Meckel-Gruber syndrome and the wpk rat. Nat Genet. 2006 Feb; 38(2):191-6. Epub 2006 Jan 15.

2005. Morris MR, Gentle D, Abdulrahman M, Maina EN, Gupta K, Banks RE, Wiesener MS, Kishida T, Yao M, Teh B, Latif F, Maher ER Tumor suppressor activity and epigenetic inactivation of hepatocyte growth factor activator inhibitor type 2/SPINT2 in papillary and clear cell renal cell carcinoma. Cancer Res. 2005 Jun 1; 65(11):4598-606.

2005. Maina EN, Morris MR, Zatyka M, Raval RR, Banks RE, Richards FM, Johnson CM, Maher ER. Identifi-cation of novel VHL target genes and relationship to hypoxic response pathways. Oncogene. 2005 Jun 30; 24(28):4549-58.

2005. Aligianis IA, Johnson CA, Gissen P, Chen D, Hampshire D, Hoffmann K, Maina EN, Morgan NV, Tee L, Morton J, Ainsworth JR, Horn D, Rosser E, Cole TR, Stolte-Dijkstra I, Fieggen K, Clayton-Smith J, Megarbane A, Shield JP, Newbury-Ecob R, Dobyns WB, Graham JM Jr, Kjaer KW, Warburg M, Bond J,

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Trembath RC, Harris LW, Takai Y, Mundlos S, Tannahill D, Woods CG, Maher ER. Mutations of the catalytic subunit of RAB3GAP cause Warburg Micro syndrome. Nat Genet. 2005 Mar; 37(3):221-3.

2004. Maina E, Morris MR, Morgan NV, Gentle D, Astuti D, Moch H, Kishida T, Yao M, Schraml P, Richards FM, Latif F, Maher ER. Molecular genetic analysis of FIH-1, FH, and SDHB candidate tumour suppressor genes in renal cell carcinoma. J Clin Pathol. 2004 Jul; 57(7):706-11.

17. Referees:

Professor Laila O. AbubakarHead of Marine Biology Research groupDepartment of BiochemistryChiromo Campus,University of NairobiTel: +254 722760070labubakar@uonbi.ac.ke

Professor Paul J. Thornalley,Protein Damage and Systems Biology Research Group,Clinical Sciences Research Institute,Warwick Medical School,University of Warwick,University Hospital,Coventry CV2 2DX, U.KP.J.Thornalley@warwick.ac.ukTel /Fax 024 7696 8594 Mobile +447884331407PA: Esther Carter Tel 02476 968665

Professor Anthony HollandHead of Learning Disabilities Research GroupUniversity of Cambridge,18b Trumpington Road, Cambridge CB2 2AH, U.KTel: +44 1223 336098ajh1008@cam.ac.uk

Mr. Davis Njuguna KanahNational Chairman- NEPHAK BoardP.O Box 75654-00200,Nairobi, Kenya,Tel: +254023875917 / 3862271Mobile: 0726164790 / 0735021241davieskanah@gmail.com

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Published Abstracts

1. African Journal of Pharmacology and Therapeutics Vol. 1 No. 3 Pages 84-91, 2012

Investigation of Prader-Willi-like Phenotype using a Whole Genome Array.

Esther N. Maina , Tessa Webb , John R Arrand , Joyce Whittington , Sarita Soni, Harm Boer , David Clarke , Anthony J. Holland

*Corresponding author: Department of Biochemistry, School of Medicine, University of Nairobi, P.O Box 3019700100, Kenya; Tel: +254-718-733987; Email: njokimaina@uonbi.ac.ke

Background: Prader-Willi syndrome (PWS) is a complex human genetic disease that arises from lack of expression of paternally inherited imprinted genes on chromosome 15q11-q13.Prader-Willi syndrome.

Objective: To use whole genome array to investigate observed Prader-Willi phenotype by assaying differential gene expression patterns in Prader-Willi like subjects.

Methodology: Combined clinical and laboratory study. Three people who participated in a large study of Prader-Willi syndrome (PWS) were found to satisfy the criteria for a firm clinical diagnosis of the syndrome using the accepted consensus scores. A score of 8 is considered to be diagnostic for PWS but despite all three scoring >8, they were genetically negative for PWS. By using Affymetrix Cytogenetics Whole-Genome 2.7M arrays which identify both loss and gain in genomic DNA and also report loss of heterozygosity regions in which all three participants showed the same genomic abnormality were determined. By comparing these regions with the UCSC human genome database, a list of potential candidate genes was compiled in which the participants had all shown the same change. Confirmation of altered gene expression was sought using qPCR to study transcription levels in each of the genes identified.

Results: Three people who participated in the study showed both maternal and paternal bands after methylationspecific PCR and they all expressed SNRPN. Increase in copy number with concomitant elevated transcription levels were found in SGSM2 which had previously been associated with severe obesity and in the protein-folding gene PPIF.

Conclusion: Over-expression caused by duplication may be a contributing factor to the PWS-like phenotype in these people.

Key words: PWS phenotype, microarray, copy-number, transcription

2. Your Doctor Magazine 2012 Jul; 4 (5-6)

Looking to the study of genes for cancer diagnoses and Treatment. (Invited mini-review)

Maina E.N.M.

Determining what causes cancer is complex. Many things are known to increase the risk of cancer, including tobacco use, certain infections, radiation, lack of physical activity, obesity, and environmental pollutants. These can directly damage genes or combine with existing genetic faults within cells to cause the disease. Approximately five to ten percent of cancers are entirely hereditary.

Research about cancer causes focuses on the following issues:•Agents (e.g. viruses) and events (e.g. mutations) which cause or facilitate genetic changes in cells destined

to become cancer.•The precise nature of the genetic damage, and the genes which are affected by it.•The consequences of those genetic changes on the biology of the cell, both in generating the defining

properties of a cancer cell, and in facilitating additional genetic events which lead to further progression of the cancer.

http://www.yourdoctormagazine.co.ke/index.php/medical-news/80-looking-to-the-study-of-genes-for-cancer-diagnoses-and-treatment?showall=1&limitstart

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3. J Pathol. 2009 Feb;217(3):345-52. doi: 10.1002/path.2487.

The ATM tumour suppressor gene is down-regulated in EBV-associated nasopharyngeal carcinoma.

Bose S, Yap LF, Fung M, Starzcynski J, Saleh A, Morgan S, Dawson C, Chukwuma MB, Maina E

, Buettner M, Wei W, Arrand J, Lim PV, Young LS, Teo SH, Stankovic T, Woodman CB, Murray PG.

Source: Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK.

AbstractA micro-array analysis using biopsies from patients with EBV-positive undifferentiated nasopharyngeal

carcinoma (NPC) and from cancer-free controls revealed down-regulation of tumour suppressor genes (TSG) not previously associated with this disease; one such gene was the ataxia telangiectasia mutated (ATM) gene. Q-PCR confirmed down-regulation of ATM mRNA and ATM protein expression in tumour cells was weak or absent in almost all cases. In NPC cell lines, however, ATM was down-regulated only in the EBV-positive line, C666.1, and in none of five EBV-negative lines. In vitro infection of EBV-negative NPC cell lines with a recombinant EBV was followed by the down-regulation of ATM mRNA and protein, and only EBV-positive cells showed a defective DNA damage response following gamma-irradiation. Our data suggest that loss of ATM function could be an important step in the pathogenesis of NPC, and may have implications for the treatment of this disease.

4. Blood. 2009 Jan 1;113(1):117-26. doi: 10.1182/blood-2008-03-142950. Epub 2008 Oct 21.

Stratification of pediatric ALL by in vitro cellular responses to DNA double-strand breaks provides insight into the molecular mechanisms underlying clinical response

Marston E, Weston V, Jesson J, Maina E, McConville C, Agathanggelou A, Skowronska A, Mapp K, Sameith K, Powell JE, Lawson S, Kearns P, Falciani F, Taylor M, Stankovic T.

Source: Cancer Research UK Institute for Cancer Studies, Birmingham University, Vincent Drive, Edgbaston, Birmingham, UK.

Abstract

The molecular basis of different outcomes in pediatric acute lymphoblastic leukemia (ALL) remains poorly understood. We addressed the clinical significance and mechanisms behind in vitro cellular responses to ionizing radiation (IR)-induced DNA double-strand breaks in 74 pediatric patients with ALL. We found an apoptosis-resistant response in 36% of patients characterized by failure to cleave caspase-3, -7, -9, and PARP1 by 24 hours after IR and an apoptosis-sensitive response with the cleavage of the same substrates in the remaining 64% of leukemias. Resistance to IR in vitro was associated with poor early blast clearance at day 7 or 15 and persistent minimal residual disease (MRD) at day 28 of induction treatment. Global gene expression profiling revealed abnormal up-regulation of multiple prosurvival pathways in response to IR in apoptosis-resistant leukemias and differential posttranscriptional activation of the PI3-Akt pathway was observed in representative resistant cases. Importantly, pharmacologic inhibition of selected prosurvival pathways sensitized apoptosis-resistant ALL cells to IR in vitro. We suggest that abnormal prosurvival responses to DNA damage provide one of the mechanisms of primary resistance in ALL, and that they should be considered as therapeutic targets in children with aggressive disease.

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5. Am J Med Genet A. 2008 Apr 1;146(7):843-53. doi: 10.1002/ajmg.a.32212.

In search of the psychosis gene in people with Prader-Willi syndrome.

Webb T, Maina EN, Soni S, Whittington J, Boer H, Clarke D, Holland A.Source: University Department of Medical and Molecular Genetics, Birmingham Women's Hospital

Edgbaston, Birmingham, UK.

AbstractThe two main causes of Prader-Willi syndrome (PWS) are a paternally derived deletion in the maternally

imprinted 15q11-q13 region or UPD(15)mat. Both mechanisms result in a loss of the active paternal contribution to the region. The affective psychosis associated with PWS has been found to be mainly confined to the propositi with UPD(15)mat rather than to those with a deletion. This suggests that the psychosis may be related to the presence of two copies rather than a single copy of a gene or genes located in the distal half of the region which is paternally imprinted, but maternally active, and whose loss results in Angelman syndrome (AS). A large population-based study of PWS allowed the identification of 12 people with a 15q11-q13 deletion who had suffered psychotic episodes and four adults with UPD(15)mat who so far had not. When these people were investigated using microsatellite markers, the 12 with a deletion were found to have two maternally derived copies of a narrow region between D15S975 and D15S661 making them effectively disomic for these loci. Thus all of the people with psychosis had two active copies of any imprinted genes in the region while all non-psychotic people (including controls) had only one. Quantitative RT-PCR studies suggest that a lack of expression of FLJ33332, either as a result of or resulting in gene dysregulation, may be associated with psychosis in PWS.

6. Psychol Med. 2008 Oct;38(10):1505-14. doi: 10.1017/S0033291707002504. Epub 2008 Jan 4.

The phenomenology and diagnosis of psychiatric illness in people with Prader-Willi syndrome.

Soni S, Whittington J, Holland AJ, Webb T, Maina EN, Boer H, Clarke D.Source: Section of Developmental Psychiatry, Department of Psychiatry, University of Cambridge, UK.

Sarita.Soni@ggc.scot.nhs.uk

AbstractBACKGROUND: Psychotic illness is strongly associated with the maternal uniparental disomy (mUPD) genetic subtype of

Prader-Willi syndrome (PWS), but not the deletion subtype (delPWS). This study investigates the clinical features of psychiatric illness associated with PWS. We consider possible genetic and other mechanisms that may be responsible for the development of psychotic illness, predominantly in those with mUPD.

METHOD: The study sample comprised 119 individuals with genetically confirmed PWS, of whom 46 had a history of

psychiatric illness. A detailed clinical and family psychiatric history was obtained from these 46 using the PAS-ADD, OPCRIT, Family History and Life Events Questionnaires.

RESULTS: Individuals with mUPD had a higher rate of psychiatric illness than those with delPWS (22/34 v. 24/85,

p<0.001). The profile of psychiatric illness in both genetic subtypes resembled an atypical affective disorder with or without psychotic symptoms. Those with delPWS were more likely to have developed a non-psychotic depressive illness (p=0.005) and those with mUPD a bipolar disorder with psychotic symptoms (p=0.00005). Individuals with delPWS and psychotic illness had an increased family history of affective disorder. This was confined exclusively to their mothers.

CONCLUSIONS: Psychiatric illness in PWS is predominately affective with atypical features. The prevalence and possibly the

severity of illness are greater in those with mUPD. We present a 'two-hit' hypothesis, involving imprinted genes on chromosome 15, for the development of affective psychosis in people with PWS, regardless of genetic subtype.

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7. J Hum Genet. 2007;52(4):297-307. Epub 2007 Jan 30.

Analysis of candidate imprinted genes in PWS subjects with atypical genetics: a possible inactivating mutation in the SNURF/SNRPN minimal promoter.

Maina EN, Webb T, Soni S, Whittington J, Boer H, Clarke D, Holland A.Source: University Department of Medical and Molecular Genetics, Birmingham Women's Hospital

Edgbaston, Birmingham, UK.

AbstractPrader-Willi syndrome (PWS) is a neurodevelopmental disorder associated with abnormalities of

chromosome 15q11q13. The majority of cases result either from a deletion approximately 4 Mb in size, affecting chromosome 15 of paternal origin or from UPD(15)mat; these account for approximately 70 and approximately 20-25% of PWS cases, respectively. In the remaining 3-5% of PWS cases where neither the deletion nor UPD is detectable, PWS is thought to be caused either by a defect in the imprinting centre resulting in a failure to reset the paternally inherited chromosome 15 derived from the paternal grandmother or, very occasionally, from a balanced translocation involving a breakpoint in 15q11q13. Nine probands with a firm clinical diagnosis of PWS but who had neither a typical deletion in the PWS region nor UPD(15)mat were investigated for inactivating mutations in 11 genes located in the PWS region, including SNURF and SNRPN, which are associated with the imprinting centre. Other genes studied for mutations included MKRN3, NDN, IPW, HBII-85, HBII-13, HBII-436, HBII-438a, PAR1 and PAR5. A possibly inactivating mutation in the SNRPN minimal promoter region was identified. No other inactivating mutations were found in the remainder of our panel of PWS subjects with atypical genetics. Expression levels of several of the candidate genes for PWS were also investigated in this series of probands. The results indicate that PWS may result from a stochastic partial inactivation of important genes.

8. J Intellect Disabil Res. 2007 Jan;51(Pt 1):32-42.

The course and outcome of psychiatric illness in people with Prader-Willi syndrome: implications for management and treatment.

Soni S, Whittington J, Holland AJ, Webb T, Maina E, Boer H, Clarke D.Source: Section of Developmental Psychiatry, Department of Psychiatry, University of Cambridge, Cambridge,

UK. ss507@medschl.cam.ac.uk

AbstractBACKGROUND: This study is part of a larger UK-wide study investigating psychiatric illness in people with Prader-Willi

syndrome (PWS), and describes the longitudinal aspect of psychiatric illness, in particular psychotic illness, and examines the use and role of psychotropic medication.

METHOD: A total of 119 individuals with genetically confirmed PWS were included in the study. An informant-based

questionnaire was administered for each participant to screen for a history of psychopathology. Those who screened positive were visited at their homes to obtain further information. This assessment included a full psychiatric history and mental state examination using the Psychiatric Assessment Schedule for Adults with Developmental Disability and the Operational Criteria Checklist for psychotic and affective illness to collect information regarding phenomenology and course of illness, and a modified life events questionnaire. At the end of the study period, informant-based telephone interviews were again carried out, up to 2.5 years after the initial screening. Information regarding medication usage was collected.

RESULTS: The results confirm previous findings that psychiatric illness in people with PWS resembles an affective

disorder. Individuals with the maternal uniparental disomy genetic subtype had a more severe course of illness than those with the deletion genetic subtype in terms of a greater risk of recurrence, more episodes, higher incidence and a possibly poorer response to medication with more side-effects. Individuals with a recurrent episode during the follow-up period had a poorer course of illness.

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Selective serotonin reuptake inhibitor medication is frequently used, and beneficial effects may reflect fundamental pathological processes in PWS. Mood-stabilizing medication was found to be of little benefit and reasons for this are examined.

CONCLUSION: The longitudinal course of psychiatric illness and response to medication in people with PWS is fully

described. Further research is needed regarding the effect of psychotropic medications, particularly mood-stabilizing medication. These data will enable informed decisions to be made regarding management options and provide information on the possible long-term outcome of illness.

9. Oncogene. 2007 Mar 8;26(11):1661-72. Epub 2006 Sep 25.

Identification of novel VHL targets that are associated with the development of renal cell carcinoma.

Maina EN, Abdulrahman M, Morris MR, Zatyka M, Raval RR, Banks RE, Wiesener MS, Richards FM, Johnson CM, Latif F, Maher ER.

Source: Department of Medical and Molecular Genetics, University of Birmingham, The Medical School, Birmingham, UK.

Abstractvon Hippel-Lindau (VHL) disease is a dominantly inherited family cancer syndrome characterized by the

development of retinal and central nervous system haemangioblastomas, renal cell carcinoma (RCC) and phaeochromocytoma. Specific germline VHL mutations may predispose to haemangioblastomas, RCC and phaeochromocytoma to a varying extent. Although dysregulation of the hypoxia-inducible transcription factor-2 and JunB have been linked to the development of RCC and phaeochromocytoma, respectively, the precise basis for genotype-phenotype correlations in VHL disease have not been defined. To gain insights into the pathogenesis of RCC in VHL disease we compared gene expression microarray profiles in a RCC cell line expressing a Type 1 or Type 2B mutant pVHL (RCC-associated) to those of a Type 2A or 2C mutant (not associated with RCC). We identified 19 differentially expressed novel VHL target genes linked to RCC development. Eight targets were studied in detail by quantitative real-time polymerase chain reaction (three downregulated and five upregulated by wild-type VHL) and for six genes the effect of VHL inactivation was mimicked by hypoxia (but hypoxic-induction of smooth muscle alpha-actin 2 was specific for a RCC cell line). The potential role of four RCC-associated VHL target genes was assessed in vitro. NB thymosin beta (TMSNB) and proteinase-activated receptor 2 (PAR2) (both downregulated by wt pVHL) increased cell growth and motility in a RCC cell line, but aldehyde dehydrogenase (ALDH)1 and ALDH7 had no effect. These findings implicate TMSNB and PAR2 candidate oncogenes in the pathogenesis of VHL-associated RCC.

10. Nat Genet. 2006 Feb;38(2):191-6. Epub 2006 Jan 15.

The transmembrane protein meckelin (MKS3) is mutated in Meckel-Gruber syndrome and the wpk rat.

Smith UM, Consugar M, Tee LJ, McKee BM, Maina EN, Whelan S, Morgan NV, Goranson E, Gissen P, Lilliquist S, Aligianis IA, Ward CJ, Pasha S, Punyashthiti R, Malik Sharif S, Batman PA, Bennett CP, Woods CG, McKeown C, Bucourt M, Miller CA, Cox P, Algazali L, Trembath RC, Torres VE, Attie-Bitach T, Kelly DA, Maher ER, Gattone VH 2nd, Harris PC, Johnson CA.

Source: Section of Medical and Molecular Genetics, Division of Reproductive and Child Health, University of Birmingham Medical School, Birmingham B15 2TT, UK.

AbstractMeckel-Gruber syndrome is a severe autosomal, recessively inherited disorder characterized by bilateral

renal cystic dysplasia, developmental defects of the central nervous system (most commonly occipital encephalocele), hepatic ductal dysplasia and cysts and polydactyly. MKS is genetically heterogeneous, with three loci mapped: MKS1, 17q21-24 (ref. 4); MKS2, 11q13 (ref. 5) and MKS3 (ref. 6). We have refined MKS3 mapping to a 12.67-Mb interval (8q21.13-q22.1) that is syntenic to the Wpk locus in rat,

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which is a model with polycystic kidney disease, agenesis of the corpus callosum and hydrocephalus. Positional cloning of the Wpk gene suggested a MKS3 candidate gene, TMEM67, for which we identified pathogenic mutations for five MKS3-linked consanguineous families. MKS3 is a previously uncharacterized, evolutionarily conserved gene that is expressed at moderate levels in fetal brain, liver and kidney but has widespread, low levels of expression. It encodes a 995-amino acid seven-transmembrane receptor protein of unknown function that we have called meckelin.

11. Cancer Res. 2005 Jun 1;65(11):4598-606.

Tumor suppressor activity and epigenetic inactivation of hepatocyte growth factor activator inhibitor type 2/SPINT2 in papillary and clear cell renal cell carcinoma.

Morris MR, Gentle D, Abdulrahman M, Maina EN, Gupta K, Banks RE, Wiesener MS, Kishida T, Yao M, Teh B, Latif F, Maher ER.

Source: Cancer Research UK Renal Molecular Oncology Group, University of Birmingham, Edgbaston, United Kingdom.

AbstractFollowing treatment with a demethylating agent, 5 of 11 renal cell carcinoma (RCC) cell lines showed

increased expression of hepatocyte growth factor (HGF) activator inhibitor type 2 (HAI-2/SPINT2/Bikunin), a Kunitz-type protease inhibitor that regulates HGF activity. As activating mutations in the MET proto-oncogene (the HGF receptor) cause familial RCC, we investigated whether HAI-2/SPINT2 might act as a RCC tumor suppressor gene. We found that transcriptional silencing of HAI-2 in RCC cell lines was associated with promoter region methylation and HAI-2/SPINT2 protein expression was down-regulated in 30% of sporadic RCC. Furthermore, methylation-specific PCR analysis revealed promoter region methylation in 30% (19 of 64) of clear cell RCC and 40% (15 of 38) of papillary RCC, whereas mutation analysis (in 39 RCC cell lines and primary tumors) revealed a missense substitution (P111S) in one RCC cell line. Restoration of HAI-2/SPINT2 expression in a RCC cell line reduced in vitro colony formation, but the P111S mutant had no significant effect. Increased cell motility associated with HAI-2/SPINT2 inactivation was abrogated by treatment with extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase (MAPK) and phospholipase C-gamma inhibitors, but not by an inhibitor of atypical protein kinase C. These findings are consistent with frequent epigenetic inactivation of HAI-2/SPINT2, causing loss of RCC tumor suppressor activity and implicate abnormalities of the MET pathway in clear cell and papillary sporadic RCC. This information provides opportunities to develop novel targeted approaches to the treatment of RCC.

12. Oncogene. 2005 Jun 30;24(28):4549-58.

Identification of novel VHL target genes and relationship to hypoxic response pathways.

Maina EN, Morris MR, Zatyka M, Raval RR, Banks RE, Richards FM, Johnson CM, Maher ER.Source: Section of Medical & Molecular Genetics, Department of Paediatrics and Child Health, University of

Birmingham, The Medical School, Birmingham B15 2TT, UK.

AbstractUpregulation of hypoxia-inducible factors HIF-1 and HIF-2 is frequent in human cancers and may result from

tissue hypoxia or genetic mechanisms, in particular the inactivation of the von Hippel-Lindau (VHL) tumour suppressor gene (TSG). Tumours with VHL inactivation are highly vascular, but it is unclear to what extent HIF-dependent and HIF-independent mechanisms account for pVHL tumour suppressor activity. As the identification of novel pVHL targets might provide insights into pVHL tumour suppressor activity, we performed gene expression microarray analysis in VHL-wild-type and VHL-null renal cell carcinoma (RCC) cell lines. We identified 30 differentially regulated pVHL targets (26 of which were 'novel') and the results of microarray analysis were confirmed in all 11 novel targets further analysed by

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real-time RT-PCR or Western blotting. Furthermore, nine of 11 targets were dysregulated in the majority of a series of primary clear cell RCC with VHL inactivation. Three of the nine targets had been identified previously as candidate TSGs (DOC-2/DAB2, CDKN1C and SPARC) and all were upregulated by wild-type pVHL. The significance for pVHL function of two further genes upregulated by wild-type pVHL was initially unclear, but re-expression of GNG4 (G protein gamma-4 subunit/guanine nucleotide-binding protein-4) and MLC2 (myosin light chain) in a RCC cell line suppressed tumour cell growth. pVHL regulation of CDKN1C, SPARC and GNG4 was not mimicked by hypoxia, whereas for six of 11 novel targets analysed (including DOC-2/DAB2 and MLC2) the effects of pVHL inactivation and hypoxia were similar. For GPR56 there was evidence of a tissue-specific hypoxia response. Such a phenomenon might, in part, explain organ-specific tumorigenesis in VHL disease. These provide insights into mechanisms of pVHL tumour suppressor function and identify novel hypoxia-responsive targets that might be implicated in tumorigenesis in both VHL disease and in other cancers with HIF upregulation.

13. Nat Genet. 2005 Mar;37(3):221-3.

Mutations of the catalytic subunit of RAB3GAP cause Warburg Micro syndrome.

Aligianis IA, Johnson CA, Gissen P, Chen D, Hampshire D, Hoffmann K, Maina EN, Morgan NV, Tee L, Morton J, Ainsworth JR, Horn D, Rosser E, Cole TR, Stolte-Dijkstra I, Fieggen K, Clayton-Smith J, Mégarbané A, Shield JP, Newbury-Ecob R, Dobyns WB, Graham JM Jr, Kjaer KW, Warburg M, Bond J, Trembath RC, Harris LW, Takai Y, Mundlos S, Tannahill D, Woods CG, Maher ER.

Source: Section of Medical and Molecular Genetics, University of Birmingham, Birmingham, B15 2TT, UK.

AbstractWarburg Micro syndrome (WARBM1) is a severe autosomal recessive disorder characterized by

developmental abnormalities of the eye and central nervous system and by microgenitalia. We identified homozygous inactivating mutations in RAB3GAP, encoding RAB3 GTPase activating protein, a key regulator of the Rab3 pathway implicated in exocytic release of neurotransmitters and hormones, in 12 families with Micro syndrome. We hypothesize that the underlying pathogenesis of Micro syndrome is a failure of exocytic release of ocular and neurodevelopmental trophic factors.

14. J Clin Pathol. 2004 Jul;57(7):706-11.

Molecular genetic analysis of FIH-1, FH, and SDHB candidate tumour suppressor genes in renal cell carcinoma.

Maina E, Morris MR,Morgan NV, Gentle D, Astuti D, Moch H, Kishida T, Yao M, Schraml P, Richards FM, Latif F, Maher ER.

Source: Section of Medical and Molecular Genetics, Department of Paediatrics and Child Health, University of Birmingham, the Medical School, Birmingham B15 2TT, UK.

AbstractBACKGROUND: Overexpression of the hypoxia inducible factor 1 (HIF-1) and HIF-2 transcription factors and the consequent

upregulation of hypoxia inducible mRNAs is a feature of many human cancers and may be unrelated to tissue hypoxia. Thus, the VHL (von Hippel-Lindau) tumour suppressor gene (TSG) regulates HIF-1 and HIF-2 expression in normoxia by targeting the alpha subunits for ubiquitination and proteolysis. Inactivation of the VHL TSG in VHL tumours and in sporadic clear cell renal cell carcinoma (RCC) results in overexpression of HIF-1 and HIF-2. However, RCC without VHL inactivation may demonstrate HIF upregulation, suggesting that VHL independent pathways for HIF activation also exist. In RCC, three candidate HIF activating genes exist-FIH-1 (factor inhibiting HIF), SDHB, and FH-which may be dependent or independent of VHL inactivation.

AIMS:

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To investigate FIH-1, SDHB, and FH for somatic mutations in sporadic RCC.METHODS: Gene mutation was analysed in primary RCCs (clear cell RCCs, papillary RCCs, and oncocytomas) and RCC cell

lines. SDHB mutation analysis was performed by denaturing high performance liquid chromatography followed by direct sequencing of aberrant PCR products. FH and FIH-1 mutation analysis were performed by single stranded conformational polymorphism and direct sequencing of PCR products.

RESULTS: No mutations were identified in the three genes investigated.CONCLUSIONS: There was no evidence to suggest that somatic mutations occur in the FH, FIH-1, or SDHB TSGs in sporadic

RCCs.