Aanvraag Minimaal / Application Minimal Dossier nummer / Dossier number: 40-00506-98-12023 DEFINITIEF

Algemene gegevens / General Information Programma / Programme

:

Investeringen Groot en Middelgroot

Subsidieronde / Subsidy round

:

Investeringen Middelgroot 2011-2012

Projecttitel / Project title

:

Building an innovative infrastructure to support a multidisciplinary approach in (inter)national large-scale research in the field of mood disorders and endocrine functioning

Projecttaal / Project language

:

Engels / English

Geplande startdatum / Planned start date

:

01-04-2012

Geplande duur / Planned duration

:

36 maanden / months

Datum indienen / Date of application

:

05-09-2011

Projecttype / Project type

:

Strategisch onderzoek / Strategic research

Vervolg eerder ZonMw-project / Continuation previously funded project ZonMw

:

Nee / No

Aanvrager / Applicant Dr. E.F.C. van Rossum MD PhD T: +31-6-54217411 | F: | E: [email protected] Erasmus MC Interne Geneeskunde Postbus 2040 3000 CA ROTTERDAM

Projectleden / Project members Dr. E.L.T. van den Akker (Mede aanvrager) Functie / Position: kinderarts-endocrinoloog | Opleiding / Education: Studierichting / Subject: T: +31 10 7036779 | F: | E: [email protected] Erasmus MC Laboratorium Inwendige Geneeskunde Postbus 2040 3000 CA ROTTERDAM

Prof. dr. E. Hoencamp (Mede aanvrager) Functie / Position: hoogleraar psychiatrie | Opleiding / Education: Studierichting / Subject: T: 070 3918190 | F: | E: [email protected] PsyQ Lijnbaan 4 2512 VA DEN HAAG

Prof. dr. F.H. de Jong (Mede aanvrager) Functie / Position: hoogleraar endocrinologie/biochemicus | Opleiding / Education: Studierichting / Subject: T: +(31)10 7030097 | F: | E: [email protected] Erasmus MC Laboratorium Inwendige Geneeskunde Postbus 2040 3000 CA ROTTERDAM

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Aanvraag Minimaal / Application Minimal Dossier nummer / Dossier number: 40-00506-98-12023 DEFINITIEF

Dr. F.J.W. Koper (Mede aanvrager) Functie / Position: biochemicus | Opleiding / Education: Studierichting / Subject: T: +(31)10 7035967 | F: +(31)10 7035430 | E: [email protected] Erasmus MC Laboratorium Inwendige Geneeskunde Postbus 2040 3000 CA ROTTERDAM

Prof. dr. S.W.J. Lamberts (Mede aanvrager) Functie / Position: hoogleraar interne geneeskunde | Opleiding / Education: Studierichting / Subject: T: +(31)10 7035967 | F: | E: [email protected] Erasmus MC Interne Geneeskunde Postbus 2040 3000 CA ROTTERDAM

Prof. dr. B.W.J.H. Penninx (Bestuurlijk verantwoordelijke) Functie / Position: directeur NESDA/prof psychiatrische epidemiologie | Opleiding / Education: Studierichting / Subject: T: +31 20 7885437 | F: | E: [email protected] VU medisch centrum / GGZ inGeest Arent Janszoon Ernststraat 887 1081 HL AMSTERDAM

Dr. Y.B. de Rijke (Mede aanvrager) Functie / Position: hoofd Diagnostisch Laboratorium Endocrinologie | Opleiding / Education: Studierichting / Subject: T: +31 10 7036625 | F: | E: Erasmus MC Klinische Chemie Postbus 2060 3000 CB ROTTERDAM

Dr. E.F.C. van Rossum MD PhD (Projectleider en penvoerder) Functie / Position: internist-endocrinoloog | Opleiding / Education: Studierichting / Subject: T: +31-6-54217411 | F: | E: [email protected] Erasmus MC Interne Geneeskunde Postbus 2040 3000 CA ROTTERDAM

Drs. A.T. Spijker (Mede aanvrager) Functie / Position: psychiater | Opleiding / Education: Studierichting / Subject: T: 070 3918190 | F: | E: [email protected] PsyQ Lijnbaan 4 2512 VA DEN HAAG

Prof. dr. F.G. Zitman (Mede aanvrager) Functie / Position: afdelingshoofd/hoogleraar psychiatrie | Opleiding / Education: Studierichting / Subject: T: 071-5269111 | F: | E: Leiden University Medical Centre Psychiatry Albinusdreef 2 2333 ZA LEIDEN

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Aanvraag Minimaal / Application Minimal Dossier nummer / Dossier number: 40-00506-98-12023 DEFINITIEF

Projectgegevens / Project information Samenvatting / Summary We propose to build a technical infrastructure to measure long-term steroid hormones, in particular the stress hormone cortisol in a unique, novel way to facilitate large-scale research with a focus on stress hormones and mood disorders. Recently we developed a technique to measure cortisol in scalp hair [1]. The advantages of this new cortisol measurement are: reliable, non-invasive and reflecting cortisol exposure of longer periods of time (months to years depending on the length of hair). This is in contrast to the currently used measurements of cortisol in serum, saliva or urine, which reflect only one time point and are dependent on the circadian rhythm and pulsatile secretion of cortisol, as well as the daily variability caused by e.g. infection or acute stress, yielding inconsistent results [2]. We aim to determine the exact relationship between long-term cortisol exposure, as measured in hair, and mood disorders. Evaluating this individual cortisol status may improve diagnosis (e.g. classification of subtypes) and predict course of disease. Ultimately, this may enable novel, tailor-made treatments for individual patients. Until now we used a modified ELISA to measure hair cortisol, but in order to enable measuring large numbers of samples and use the hair extracts for multiple purposes a liquid chromatography tandem mass spectrometre (LC-MS/MS) is of great value and is therefore part of our proposal for an infrastructure. We intend to use the LC-MS/MS to apply our recently developed method to measure steroid hormones for our project which concerns a collaborative initiative of the multi-centric Netherlands Study of Depression and Anxiety (VUMC, LUMC, UMCG), PsyQ (The Hague) and the Department of Internal Medicine, Laboratory of Endocrinology, Erasmus MC, Rotterdam. Importantly, in addition, we aim to set up a national biobank of scalp hair extracts, which will facilitate other independent research groups to obtain data on long-term steroid hormones in scalp hair, measured in our endocrine laboratory. Cortisol plays an important role in numerous pathological conditions, e.g. obesity, diabetes mellitus, hypertension, hypercholesterolemia, cardiovascular disease, infectious disease, auto-immune disease, as well as cognitive functioning and dementia. This makes this new long-term cortisol measurement of interest for a wide range of research areas. In the past months we received numerous international and national requests from various medical disciplines to measure hair cortisol levels in large epidemiological cohort studies or in patients groups, which exceeds our current limited capacity. By setting up this new infrastructure of a biobank of hair extracts and performing steroid hormone measurements in hair samples we can contribute to strengthen the leading position the Netherlands has in this quickly developing endocrine field.

Bijzondere gegevens / Additional information Vergunningen / Permits Verklaring nodig / Statement required? Ja / Yes METC DEC WBO

Nee / No

X

Status verklaring / Statement status Nog niet Verkregen / Aangevraagd / aangevraagd / Not Acquired Applied applied yet X

X X

X

Onderschrijvingen / Assents Ja / Yes

Nee / No

Code biosecurity / Code Biosecurity Code openheid dierproeven / Code Transparency of Animal Testing

N.v.t. / N.A. X X

Andere vergunningen / Other permits

Ondertekening / Signatures Naam projectleider en penvoerder: E.F.C. van Rossum

Naam bestuurlijk verantwoordelijke: B.W.J.H. Penninx

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Aanvraag Minimaal / Application Minimal Dossier nummer / Dossier number: 40-00506-98-12023 DEFINITIEF Plaats en datum:

Plaats en datum:

Handtekening:

Handtekening:

Aangemaakt door ProjectNet / Generated by ProjectNet: 05-09-2011 14:44

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Application Form Investment Subsidy NWO Medium 2011/2012

Please indicate which NWO Division should review your application (several options possible). ALW Ο

CW Ο

EW Ο

1. Applicants(s) a. Principal applicant / contact Name, title(s): University: Research school: E-mail: b.

c

GW Ο

MaGW Ο

ZonMw X

Dr. E.F.C. van Rossum, MD PhD Erasmus Medical Center molecular medicine/dept internal medicine, section endocrinology [email protected]

Female

Co-applicant(s) / contact Name, title(s): Prof. dr. B.W.J.H. Penninx, MD PhD University: VU University Medical Center/GGZ inGeest. Cross-appointment with Leiden University Medical Center and University Medical Center Groningen Research school: EMGO+/ Neuroscience Campus Amsterdam E-mail: [email protected] Composition research team Name and title Specialization

Role

Prof. dr. Frans Zitman, MD PhD Prof. dr. Erik Hoencamp, MD PhD Drs. Annet T. Spijker, MD

Psychiatry

Co-investigator

Psychiatry

Co-investigator

Psychiatry

Co-investigator

Dr. Jan W. Koper, PhD

Biochemistry

Co-investigator and technical expert

Drs. Laura Manenschijn, MD

Medicine

PhD student

Prof. dr. Steven W.J. Lamberts, MD PhD

Internal Medicine /Endocrinology

Co-investigator

NWO Application form for medium sized investments 2011/2012

Female

Employment/instit ute Leiden University Medical Center Leiden University, PsyQ, The Hague Division of mood disorders, PsyQ, The Hague Laboratory of Endocrinology, Erasmus MC, Rotterdam Internal Medicine, Erasmus MC, Rotterdam Erasmus MC, Rotterdam page 1

d

Prof. dr. Frank H. de Jong, PhD Dr. Yolanda B. de Rijke, PhD

Endocrinology

Dr. Erica.L.T.van den Akker, MD PhD

pediatric endocrinology

Clinical Chemistry

Co-investigator, Expert on steroids Co-investigator and technical expert Co-investigator

Erasmus MC, Rotterdam Erasmus MC, Rotterdam Erasmus MCSophia, Rotterdam

Does the local authority support your application? X Yes Ο No (Does your institute/university accept the conditions for support by NWO / ZonMw ?) Relevant authority: Name: Prof.dr. C.P.Verrijzer Position: Chair MRace Apparatuur commissie

2. Project title Building an innovative infrastructure to support a multidisciplinary approach in (inter)national large-scale research in the field of mood disorders and endocrine functioning 3. Project summary (max. half page) We propose to build a technical infrastructure to measure long-term steroid hormones, in particular the stress hormone cortisol in a unique, novel way to facilitate large-scale research with a focus on stress hormones and mood disorders. Recently we developed a technique to measure cortisol in scalp hair [1]. The advantages of this new cortisol measurement are: reliable, non-invasive and reflecting cortisol exposure of longer periods of time (months to years depending on the length of hair). This is in contrast to the currently used measurements of cortisol in serum, saliva or urine, which reflect only one time point and are dependent on the circadian rhythm and pulsatile secretion of cortisol, as well as the daily variability caused by e.g. infection or acute stress, yielding inconsistent results [2]. We aim to determine the exact relationship between long-term cortisol exposure, as measured in hair, and mood disorders. Evaluating this individual cortisol status may improve diagnosis (e.g. classification of subtypes) and predict course of disease. Ultimately, this may enable novel, tailor-made treatments for individual patients. Until now we used a modified ELISA to measure hair cortisol, but in order to enable measuring large numbers of samples and use the hair extracts for multiple purposes a liquid chromatography tandem mass spectrometre (LC-MS/MS) is of great value and is therefore part of our proposal for an infrastructure. We intend to use the LC-MS/MS to apply our recently developed method to measure steroid hormones for our project which concerns a collaborative initiative of the multi-centric Netherlands Study of Depression and Anxiety (VUMC, LUMC, UMCG), PsyQ (The Hague) and the Department of Internal Medicine, Laboratory of Endocrinology, Erasmus MC, Rotterdam. Importantly, in addition, we aim to set up a national biobank of scalp hair extracts, which will facilitate other independent research groups to obtain data on long-term steroid hormones in scalp hair, measured in our endocrine laboratory. Cortisol plays an important role in numerous pathological conditions, e.g. obesity, diabetes mellitus, hypertension, hypercholesterolemia, cardiovascular disease, infectious disease, auto-immune disease, as well as cognitive functioning and dementia. This makes this new long-term cortisol measurement of interest for a wide range of research areas. In the past months we received numerous international and national requests from various medical disciplines to measure hair cortisol levels in large epidemiological cohort studies or in patients groups, which exceeds our current limited capacity. By setting up this new infrastructure of a biobank of hair extracts and performing steroid hormone measurements in hair samples NWO Application form for medium sized investments 2011/2012

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we can contribute to strengthen the leading position the Netherlands has in this quickly developing endocrine field. 4a. Keywords (max. 5) cortisol, steroids, scalp hair, depression, biobank 4b. Text for press release (to be used if your application is granted) Biobank voor hormoonmetingen in hoofdhaar Dr E.F.C. [Liesbeth] van Rossum, V, 15 juli 1975, Erasmus MC, Afdeling Inwendige Geneeskunde/Endocrinologie Depressie is een veel voorkomende ziekte, waarbij het stresshormoon cortisol een belangrijke rol vervult. In dit onderzoek wordt de relatie tussen cortisol en stemmingsstoornissen onderzocht, middels nieuwe testen op hoofdhaar die voor het eerst de blootstelling aan cortisol over langere tijd meten.

5. Is this a revised application? Yes, in 2011, NWO-MaGW filenumber: 480-11-003 (see item 6) 6. Has this application been submitted elsewhere? The application is not currently submitted elsewhere, but it has been submitted with NWOMaGW in May 2011. The project has been evaluated positively (scientific quality and interest, innovation, public interest, potential to lead to new insights), but the scope was found to be too medically oriented for the field of NWO-MaGW. We were advised to submit it to ZonMW–middelgroot.

7. Detailed description of the research and investment plan The prevalence of mood disorders in the Netherlands is high (around 7%) and poses an immense burden on health care costs. The most consistent biological finding in patients with unipolar or bipolar depression is a hyperactivity of the hypothalamic-pituitary-adrenal (HPA)-axis [3]. This axis is part of the physiological stress system. Stress often precedes a major depressive episode and predicts a worse treatment outcome [4]. The main stress hormone in humans is cortisol. In addition to Glucocorticoid Receptor (GR) resistance, also increased GR activation may promote depressive symptoms. In Cushing’s syndrome, characterized by severe hypercortisolism, depressive symptoms occur frequently. Not only glucocorticoid resistance seems to be related to depression, but also enhanced GR effects in limbic brain regions may contribute to the development of depression. The role of the stress hormone cortisol in depressive disorders is not clear yet, since cortisol measurements used in the past years yielded conflicting results [2]. Assessment of long term cortisol levels by a novel biomarker Evidence is mounting that cortisol measurements using serum, salivary or urine, fail to adequately reflect cortisol’s true impact because of the circadian rhythm of cortisol levels (high in the morning, lower during the afternoon, and lowest at night) and the pulsatile way in which cortisol is secreted [2, 5]. Also, large daily variations due to e.g. acute stress or infection are an important limitation of the traditional ways of measuring cortisol levels. A novel and non-invasive parameter is measuring cortisol in scalp hair [1, 6, 7]. Hair grows with an average of 1 cm per month, and it has been shown that cortisol can be reliably measured in hair. The use of hair provides the opportunity to measure long term cortisol levels (reflecting mean levels of the past months) in an easy way without limitations caused by the pulsatility and circadian rhythm of cortisol or acute circumstances [8]. Since the method is simple and non-invasive it is very well suitable for large-scale epidemiological studies. Hair cortisol can be used to obtain a robust parameter reflecting mean free cortisol levels over extended periods of time [9-11]. At present no NWO Application form for medium sized investments 2011/2012

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large-scale studies have been performed using this retrospective index of cortisol secretion over a prolonged period of time. Major field of interest: Long-term cortisol levels and mood disorder. Stress is often a trigger for a new episode of mood disturbances. However, the relation between chronically elevated stress hormone levels and the development of mood disorders is unknown, since until now only cortisol measurements were available which reflect levels during the past minutes to hour(s). Using the novel biomarker of hair cortisol we will be able to detect cortisol levels retrospectively over longer periods of time (months to years, depending on the length of the hair strands). This enables for the first time the determination of the exact relationship between cortisol and mood disorders in large cohorts. We will study associations with diagnostics, course of mood disorders, response to treatment, cognitive impairments, as well as with somatic diseases (in particular metabolic syndrome, an important health problem in psychiatry, which also seems to be related to elevated cortisol levels). This may provide new insights into the role of cortisol in mood disorders, possibly offering new tools to diagnose different subtypes of depression and develop new treatment strategies based on an individual’s cortisol status. Study populations 1. Netherlands Study of Depression and Anxiety (NESDA): an eight-year longitudinal cohort study among 2981 participants aged 18 through 65 years. The sample consists of 1701 persons with a current diagnosis of depression and/or anxiety disorder, 907 persons with life-time diagnoses or at risk because of a family history or sub threshold depressive or anxiety symptoms, and 373 healthy controls. Data are available on psychiatric diagnoses, symptom severity and duration, personality traits, family history, depressive and anxiety vulnerability cognitions, childhood and recent life events and trauma during all conducted assessments, as well as genetic data for a Genome Wide Association Study and information about somatic diseases and metabolic abonormalities (e.g. metabolic syndrome). This will allow for a detailed investigation on various psychiatric, somatic and environmental characteristics in association with hair steroid levels. 2. Bipolar Disorder Study (PsyQ, The Hague): a two-year longitudinal study in 326 outpatients with bipolar disorder, aged 18 years and over. Data are available on psychiatric diagnosis, age of onset of first symptoms, the number of episodes of (hypo)mania and depression. The presence of co-morbid anxiety disorders (according DSM-IV-TR) was assessed with the Mini-International Neuropsychiatric Interview (MINI). Psychiatric co-morbidity was assessed by MINI and included all lifetime and current anxiety disorders, somatoform disorders, eating disorders, substance disorders, suicidality and premenstrual dysphoria. In addition physical data and diseases have been registered. Description of the investment The proposed investment concerns an infrastructure to set up a bio-bank of scalp hair extracts, including a liquid chromatography–tandem mass spectrometre (LC-MS/MS), as well as personnel. 1. Biobank of scalp hair and hair extracts Hair samples consisting of a minimum 10 mg of hair (ca 100 strands) will be collected from the posterior vertex of the patients of whom numerous psychiatric and somatic parameters already have been documented. An overnight methanol extraction of fragmented hair samples will be performed. After incubation, the samples will be further processed. Of each extract a small amount (approximately 10%) will be used to measure cortisol levels using LC-MS/MS. The major part of the hair extract will be kept in storage. Importantly, this enables extensive additional research of other steroid hormones for future research, as well as measurement of steroid hormone metabolites (and thus steroid metabolism) and potentially also future measures of interest e.g. substrates as medications, toxins etc. In addition, when handling many samples it is imperative to have a good tracking system (barcode), which will be part of this infrastructure.

NWO Application form for medium sized investments 2011/2012

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2. Liquid chromatography–tandem mass spectrometre (LC-MS/MS) Currently cortisol extracted from hair is measured using an immunochemical technique, which is limited by its labour-intensive work and usage of large volumes of hair extracts. An important part of this proposal is to invest in an LC-MS/MS for the purpose of determining cortisol in hair. This technique is rapidly being recognized as state of the art. It has the advantages of increased specificity and sensitivity, and offers also the possibility of measuring several steroids, as well as steroid metabolites, simultaneously. Moreover, substantial reduction of sample volumes has previously been demonstrated and validated [12], which is a very important advantage. Reduction of the sample size of hair extracts will result in the possibility to retain a considerable portion of each sample for future projects. 3. Personnel For efficient management of the proposed facility it will be necessary to employ a laboratory technician (daily operation of the facility) and a daily supervisor/data manager for maintaining the required quality level and to keep the technical knowhow up to date. In particular the first years of operating this advanced technique are known to require an investment in time and personnel to develop the assays, maintain the processes and assure continuous quality checks. Reasons for the proposed investment We have recently started to measure cortisol in scalp hair using immunoassays with the aim of finding a parameter that reflects long term cortisol exposure, rather than the acute values that result from determinations in blood or saliva and, to a lesser extent, in urine. Following our recent scientific report [1] as well as in the lay press (see also search terms Google “stresshormoon haar”) on the results of our investigations we found that there was considerable interest in this technique from various research groups around the country. This reflects a high potential for great societal impact in various fields in the Netherlands. Since cortisol is a major factor in numerous pathological conditions, including metabolic disorders, obesity, cardiovascular disease, inflammatory and auto-immune diseases, as well as cognitive decline, this novel long-term cortisol measurement is of great importance for a wide variety of research groups. At present we are the only centre in the Netherlands which has the technique of measuring steroids in scalp hair available. Very few other laboratories in the world have currently developed this technique. In addition, other national and international research groups working in the field of steroids have shown great interest in our measurements of steroids in scalp hair. This also shows an urgent need of a high throughput technique facilitating many (inter)national research projects. Insight into the effects of corticosteroids on mood disorders may lead to the unravelling of underlying biological pathophysiological mechanisms and potentially new treatment strategies. In addition to the currently developed cortisol measurements in hair extracts we aim to measure other adrenal hormones and sex hormones in hair, and study their role in mood disorders in subsequent projects. Furthermore, we are interested in measuring steroid metabolites in hair reflecting long-term status of metabolism of steroids, which will provide further insights in the patho-physiological role of corticosteroids in mood disorders. Future projects may focus on other compounds which can be measured in scalp hair extracts, e.g. toxins and drugs. An LC-MS/MS will allow to measure all the above mentioned substances of interest with very high sensitivity. In addition, other groups involved in other research areas, including metabolic disorders, cardiovascular disease, auto-immune disorders or dementia, could greatly benefit from long-term cortisol measurements in scalp hair extracts. In view of this we formulated the current proposal which aims to offer a facility for the collection, extraction, storage and analysis of steroids in scalp hair. Since the technical and scientific expertise is already present in our laboratory we feel that building the proposed infrastructure is certainly feasible. NWO Application form for medium sized investments 2011/2012

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Future perspectives In the near future, access to this national bio-bank of hair extracts will also be provided to other independent research groups. Usage of an LC-MS/MS will provide possibilities for analyzing multiple steroid hormones simultaneously with superior quality and will allow high throughput testing using small sample volumes. The latter is important since each hair extract sample can be used for multiple purposes allowing additional future research using the same samples of large epidemiological cohort studies (e.g. measuring other steroids as testosterone, androstenedione, estradiol, or toxins, medications or other biochemical substrates which may have clinical relevance and will be of interest for numerous types of medical research). Internationally, the field of hair cortisol is relatively new and has been developed in the past year by only a few groups. In the past years the research on long-term cortisol was initiated by forensic researchers and was primarily focused on hair of monkeys. Only very recently, articles on clinical applications in humans have been published and at international congresses this innovative topic is getting increasing attention[10]. Since our group is one of the few in the world who managed to set up and validate the method, we are in an internationally leading position. Thus, in addition to future projects of our own research groups building on the present research, by means of this infrastructure we will facilitate other research groups. With this investment we build the logistics and expertise that other projects can use. In this way, we will contribute to the strong international position of Dutch medical research. Several concrete examples of current requests to our laboratory for long-term steroid measurements, stressing the necessity for a national bio bank of hair extract samples and the facilities for their analysis: - BiG Consortium N=2000, a national multi-center study on bipolar disorder - Generation R study, Rotterdam. N=5000, a prospective cohort study from fetal life until young adulthood. Dr Jaddoe, Sophia’s Children’s Hospital, Erasmus MC, Rotterdam - Adolescents at risk of anxiety and depression (Ariadne) study, Groningen, N=520. - Longitudinal Aging Study Amsterdam (LASA) study, Amsterdam, N=2000. - MOOD-INFLAME, N=425, an European consortium on inflammation and mood disorders. - Late effects of childhood abuse, N=1000, Prof B.M. Elzinga, Dept of Neuropsychology, Leiden University - Monitoring hydrocortisone therapy in children with adrenogenital syndrome, N=60 Dr. E.L.T.van den Akker, Sophia’s Children’s Hospital, Erasmus MC, Rotterdam - Hydrocortisone replacement therapy in patients with pituitary macroadenomas, N=200 Dr N. Biermasz, Endocrinology, Leiden University - Neurobiological Inheritance of Psychological Trauma, N=300, Dr IL de Castro Vale, University of Porto, Portugal - Health effects of disturbances of the circadian rhythm on cancer, metabolic syndrome and other chronic diseases. National Institute for Public Health and the Environment (RIVM), Dr A. Opperhuizen and Dr W. Roodenburg By means of this proposed infrastructure we will be directly able to meet the above mentioned demands. References 1. Manenschijn, L., Koper J.W., Lamberts S.W.J., van Rossum E.F.C. Evaluation of a method to measure long term cortisol levels. Steroids. 2011 Sep-Oct;76(1011):1032-6. 2. Knorr, U., et al., Salivary cortisol in depressed patients versus control persons: a systematic review and meta-analysis. Psychoneuroendocrinology.2010 35(9): 1275-86. 3. Holsboer, F., The corticosteroid receptor hypothesis of depression. Neuropsychopharmacology, 2000. 23(5): 477-501. NWO Application form for medium sized investments 2011/2012

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4.

5. 6. 7. 8. 9. 10. 11. 12.

Kiosses, D.N., A.C. Leon, and P.A. Arean, Psychosocial Interventions for Late-life Major Depression: Evidence-Based Treatments, Predictors of Treatment Outcomes, and Moderators of Treatment Effects. Psychiatr Clin North Am. 2011 34(2): 377401. Stetler, C. and G.E. Miller, Depression and hypothalamic-pituitary-adrenal activation: a quantitative summary of four decades of research. Psychosom Med. 2011 73(2): 114-26. Van Uum, S.H., et al., Elevated content of cortisol in hair of patients with severe chronic pain: a novel biomarker for stress. Stress, 2008. 11(6): 483-8. Davenport, M.D., et al., Analysis of endogenous cortisol concentrations in the hair of rhesus macaques. Gen Comp Endocrinol, 2006. 147(3): 255-61. D'Anna-Hernandez, K.L., et al., Hair cortisol levels as a retrospective marker of hypothalamic-pituitary axis activity throughout pregnancy: comparison to salivary cortisol. Physiol Behav. 2011 104(2): 348-53. Kirschbaum, C., et al., Hair as a retrospective calendar of cortisol productionIncreased cortisol incorporation into hair in the third trimester of pregnancy. Psychoneuroendocrinology, 2009. 34(1): 32-7. Gow, R., et al., An assessment of cortisol analysis in hair and its clinical applications. Forensic Sci Int. 2010 196(1-3): 32-7. Dettmer, A.M., et al., Physiological and behavioral adaptation to relocation stress in differentially reared rhesus monkeys: Hair cortisol as a biomarker for anxietyrelated responses. Psychoneuroendocrinology. 2011 Jun Epub ahead of print Kushnir, M.M., et al., Liquid chromatography tandem mass spectrometry for analysis of steroids in clinical laboratories. Clin Biochem. 2011 44(1): 77-88.

8. The quality and competence of the research team. In the past four years the multi-centre NESDA study yielded many (>150) international publications, in high impact journals (e.g. Nature Genetics 11/ Nature 2/Biological psychiatry 6/ Arch Gen Psych 3) showing a great significance and quality of this research group. The Bipolar Disorder study (PsyQ The Hague/LUMC) is a novel well phenotyped study cohort, which developed in a short period of time the largest cohort of bipolar patients in the Netherlands with great potential. In addition, our endocrine laboratory at the Erasmus MC has also an outstanding long-term international reputation in the field of steroid hormone research. An example of a researcher of our endocrine group with an impressive track record is Prof SWJ Lamberts with 681 international publications, 17 prestigious (inter)national awards, who is past president of the European Society of Endocrinology. The first applicant (EFC van Rossum, internist-endocrinologist since 2010) has a track record in the field of cortisol research with 50 peer-reviewed international articles, a current H-index of 21, who obtained her PhD (cum laude) in 2005, a NWO-Veni grant in 2008, and a fellowship of the Netherlands Brain Foundation (Hersenstichting) in 2011. She is currently supervising 4 PhD students. The second applicant (Prof BWJH Penninx), is principal investigator of the NESDA study, a NWO VIDI and VICI laureate, and has an excellent reputation in epidemiologic psychological and psychiatric studies, reflected in 296 publications, a current H-index of 52, and a research staff consisting of 21 PhD-students and 3 postdocs. Ten relevant publications (related to this proposal) 1. Effect of depression on cardiac mortality. Results from a community-based longitudinal study. Penninx BWJH, Beekman ATF, Honig A, Deeg DJH, Schoevers RA, van Eijk JThM, van Tilburg W. Arch Gen Psychiatr 2001;58:221-227. 2. Major Depressive Disorder and Hypothalamic-pituitary-adrenal axis activity: Results from a large cohort study.Vreeburg SA, Hoogendijk WJ, van Pelt J, de Rijk RH, Verhagen JCM, van Dyck R, Smit JH, Zitman FG, Penninx BW. Arch Gen Psychiatry 2009;66:617626. NWO Application form for medium sized investments 2011/2012

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3. Shift work at young age is associated with elevated long-term cortisol levels in scalp hair and body mass index. Manenschijn L, van Kruysbergen RG, de Jong FH, Koper JW, van Rossum EFC. J Clin Endocrinology and Metabolism, 2011 in press 4. Dimensions of depression and anxiety and the hypothalamo-pituitary-adrenal axis.Wardenaar KJ, Vreeburg SA, van Veen T, Giltay EJ, Veen G, Penninx BW, Zitman FG. Biol Psychiatry. 2011 Feb 15;69(4):366-73. 5. Evaluation of a method to measure long term cortisol levels. Manenschijn L, Koper JW, Lamberts SW, van Rossum EF. Steroids. 2011 Sep-Oct;76(10-11):1032-6. 6. The Netherlands Study of Depression and Anxiety (NESDA): rationale, objectives and methods. Penninx BW, Beekman AT, Smit JH, Zitman FG, Nolen WA, Spinhoven P, Cuijpers P, De Jong PJ, Van Marwijk HW, Assendelft WJ, Van Der Meer K, Verhaak P, Wensing M, De Graaf R, Hoogendijk WJ, Ormel J, Van Dyck R; NESDA Research Consortium.Int J Methods Psychiatr Res. 2008;17(3):121-40. 7. Glucocorticoid receptor polymorphisms in major depression. Focus on glucocorticoid sensitivity and neurocognitive functioning. Spijker AT, van Rossum EF. Ann N Y Acad Sci. 2009 Oct;1179:199-215. 8. Identifying depressive subtypes in a large cohort study: results from the Netherlands Study of Depression and Anxiety (NESDA). Lamers F, de Jonge P, Nolen WA, Smit JH, Zitman FG, Beekman AT, Penninx BW. J Clin Psychiatry. 2010 Dec;71(12):1582-9. Epub 2010 Jul 13. 9. Functional polymorphism of the glucocorticoid receptor gene associates with mania and hypomania in bipolar disorder. Spijker AT, van Rossum EF, Hoencamp E, DeRijk RH, Haffmans J, Blom M, Manenschijn L, Koper JW, Lamberts SW, Zitman FG. Bipolar Disord. 2009 Feb;11(1):95-101. 10. Polymorphisms of the glucocorticoid receptor gene and major depression. Van Rossum EF, Binder EB, Majer M, Koper JW, Ising M, Modell S, Salyakina D, Lamberts SW, Holsboer F.Biol Psychiatry. 2006 Apr 15;59(8):681-8.

9. Social relevance see also item 7 “Reasons for the proposed investment”. In addition, examples of impact to the lay public: several press releases concerning our research related to stress hormones, depression, and cardiovascular risk factors were initiated by the American Endocrine Society, Erasmus MC, and European Society of Endocrinology. These releases yielded articles in (inter)national newspapers (e.g. Washington Post 2x, Algemeen Dagblad 3x, Telegraaf 2x, NRC 1x, Nederlands Dagblad 2x, Radio interviews (5x), as well as over 100 articles on (inter)national websites.

10. National interest and access Endocrinology in the Netherlands has a very good international position. However, in order to maintain this position, continuous innovation and development is necessary. With respect to recent international developments it is obvious that the application of LC-MS/MS techniques is an absolute must to stay in the forefront. This is also clearly the long term strategy of the department. The innovative part of this project proposal is the combination of the analysis facility with the “hair bio bank” and its (inter-)national accessibility. As NWO Application form for medium sized investments 2011/2012

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outlined in point 7 (Future perspectives) there is already considerable interest from other institutions in the Netherlands, and collaboration with these institutions will be initiated. The arrangements under which these collaborations will take place may include pure collaboration, service against minimal costs, or a combination of both. see also item 7 “Reasons for the proposed investment, Future perspectives”. Data which will be obtained by means of the proposed infrastructure will be available for other researchers. In analogy to the present data from the multi-centric NESDA study, which are being used by multiple research groups inside (including researchers from Radboud University, Erasmus MC, Tilburg University, Maastricht University) and outside (e.g. by researcher from Australia, USA universities and NIH institutes) the Netherlands, we will make the long-term values obtained from hair extracts (steroid hormones and other substances) available for other research groups. Data will be provided after an analysis plan has been submitted, which can be done by any researcher. After finishing the project, data will be archived in the publically accessible Data Archiving and Networked Services (DANS).

11. Description of environment and implementation of the requested equipment Accommodation for the proposed facility includes space for the LC-MS/MS machine to be housed, working space for the operation of the LC-MS/MS and the required pre-treatment of hair samples and space for the archiving of the samples to be bio-banked. These requirements can be met within the Laboratory of Endocrinology of the Erasmus MC. The surrounding infrastructure is the Laboratory of Endocrinology, consisting of five internationally competitive research groups and the Diagnostic Laboratory of Endocrinology. Both in scientific and in technical sense, this environment will be conducive to further evolution and expansion of the proposed facility and the projects for which it will be used. Within the laboratories of the department of Internal Medicine and the department of Clinical Chemistry sufficient know how is available to support this project and the development of additional methodology. In principle the facility will be open for research projects in the field of mood disorders and endocrine functioning, as is described in the title of this proposal. Running costs will initially be carried by the department of Internal Medicine from the funds available for ongoing projects, eventually it may be necessary to ask for (minimal) contributions payable from the operating funds of the projects making use of the facility. It is anticipated that the facility will be used extensively (see future perspectives), with sufficient capacity to leave room for the development of methods for the detection of other substances (e.g. steroid metabolites). The apparatus will be part of the Erasmus MC apparatus replacement schedule, which allows for replacement after the depreciation period has expired (typically 5 – 7 yr for this type of apparatus).

12. Financial/technical aspects The costs consist of the acquisition of an LC-MS/MS machine, the cost of the hair bank facility and the personnel cost for management and operation of the facility. Additionally computer facilities are needed to develop and maintain the extensive databases. For this purpose budget for server capacity, database storage capacity and an automated tracking system for high throughput processing is included in this proposal. Personnel costs have been based on the model supplied at www.nwo.nl/contractvsnu in combination with the salary scales applicable in the Dutch UMCs as of August 1st 2011. The cost of the LCNWO Application form for medium sized investments 2011/2012

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MS/MS machine is based on a quotation from Waters for a Waters Xevo TQ-S machine, which meets the specifications needed for the proposed project.

Matching Item description

To be paid by NWO / ZonMW

whom) 2012

Liquid chromatography–mass spectrometer

(indicate by

2013

2014 100 (Erasmus MC)

380

Facilities for the storage of hair and hair extract samples, and computer facilities (hardware and software) for

20

20

20

40 (Erasmus MC)

44

44

45

50 (Erasmus MC)

the hair sample registration and storage/analysis databases Laboratory technician (HBO, 1.0 fte, 3 yrs= 183 k-euro). Processing hair samples, steroid extraction, operating LC-MS/MS, datamanagement Dr JW Koper (PhD, 0.25 fte, 3 yrs). Daily management, technical /biochemical assistance in the

P.M.

development of new assays (other

(ErasmusMC)

steroid hormones and metabolites, as well as other substances/toxins) Dr EFC van Rossum (UMS, 0.1 fte, 3 yrs) PI, clinical input on steroid

P.M.

analysis, maintaining network with

(ErasmusMC)

the collaborating psychiatrists, performing the proposed studies Subtotal (max. 75% NWO/ZonMw)

444

Total

64 573

12b. Do you expect financial support from additional source(s)? If yes, please specify: Source Hersenstichting, fellowship (The Netherlands Brain Foundation)

65

Amount (k€) 150

190 763

X Yes

O No

Requested/guaranteed Guaranteed

12c. We are aware that this grant proposal round is centred on material investments. For this reason the sum that is included in the proposal for personnel cost has been kept to a minimum. A facility such as the one proposed here is not a one-time issue. Our intention is to collect samples, with the primary aim to extract cortisol from those samples and to NWO Application form for medium sized investments 2011/2012

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determine its concentration and relate that to the (medical) history of the person who donated that sample, especially in relation to issues such as depression. Goals further afield include the possibility to analyse stored extracts for other substances in samples derived from well-defined populations with extensive documentation of their medical history. Additionally, in addition to extracts from hair, we will also store the remaining hair sample, which will be available for future extraction with novel/other methods of extraction, possibly aimed at other substances and other questions. To achieve this we need to hire a technician who collects, handles, extracts and stores the samples. Besides the labour-intensive hair processing, hair extractions and biochemical analyses this technician will also take care of the correct processing and storage of the obtained data in large databases, which is important in a high throughput setting. Therefore, we feel that this application should include personnel as well as the material investment. The proposed facility offers these possibilities and they cannot be obtained elsewhere and certainly in the initial period this necessitates dedicated attention. For this reason a specialized technician will be necessary for its daily operation and adequate input is needed from scientific staff for the daily management and further development (dr Koper, daily supervision, development of new targets, methods and priorities) and for careful clinical input and further propagation and publication of the facility in order to achieve maximal benefit from the investment (dr van Rossum).

NWO Application form for medium sized investments 2011/2012

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