Biobank Netværksmøde. 17. Marts 2016

Transkript

1 Biobank Netværksmøde 17. Marts 2016

2 PRAKTISKE INFORMATIONER Toiletter Frokost serveres her i lokalet Brug muligheden for et besøg på messen i frokostpausen eller efter mødet Alle foredragsholdere bedes aflevere en version vi kan lægge på DCB hjemmeside ( Efter frokost overtager Erik styringen af mødet og sørger for en god afrunding

3 Program - formiddag 10:00-10:10 Velkomst og praktiske informationer (Estrid Høgdall, Regionernes Biobank, Patologiafdelingen, Herlev Gentofte Hospital) 10:10-10:20 Indledning til dagens to emner præ-analytiske faktorer og SOPs (Estrid Høgdall, Regionernes Biobank, Patologiafdelingen, Herlev Gentofte Hospital) 10:20-10:50 Preanalytical considerations when collecting blood samples for biobanking (Emil Daniel Bartels, Afdelingslæge, ph.d., Klinisk Biokemisk Afdeling, Rigshospitalet) 10:50-11:20 The influence of different fixatives and preparation methods on morphology, immunohistochemistry and molecular analyses (Lone Bojesen, kvalitetskoordinator/udviklingsbioanalytiker, Patologiafdelingen, Herlev Gentofte Hospital) 11:20 11:50 Investigating potential microrna biomarkers as indicators of metastatic disease in early CRC (Rikke Karlin Jepsen, læge, ph.dstuderende, Patologiafdelingen, Herlev Gentofte Hospital) 11:50 13:00 Frokost og netværk/messebesøg

4 Program - eftermiddag 13:00 13:30 Diskussion af foredrag hvad skal vi være opmærksomme på? Hvad bør vi undersøge? Er der noget vi skal ensrette/ændre? Emner til næste netværksmøde 13:30-14:15 SOPs set med en GCP-indgangsvinkel (Charlotte Calov, Funktionsleder, cand.med.vet, GCP-enheden, Afdeling for Klinisk Biokemi og Farmakologi Odense Universitetshospital) 14:15 14:30 Afrunding på dagen de bedste take home messages arrangør for næste møde (Erik Sørensen, Molekylær Biolog. Klinisk Immunologisk Afdeling. Blodbanken og Vævstypelaboratoriet, Diagnostisk Center Rigshospitalet) God tur hjem måske efter besøg på messen

5 Dagens emner præ-analytiske faktorer og SOPs Hvorfor overhovedet???? Et par eksempler

6 Så konklusioner skal være korrekte og ikke afspejle præ-analytiske faktorers indflydelse på et analyseresultat

7 Præ-analytiske faktorer: Uregerlige kilder Kontrollerbare kilder Kilder der kan komme ind imellem Dagen skal give omtanke for hvordan man undersøger, tager højde for og vurderer effekten af præanalytiske kilder til variation

8 OVX1 Radioimmunoassay Results Are Dependent on the Method of Sample Collection and Storage, Estrid V.S. Høgdall,1* Claus K. Høgdall,2 Susanne K. Kjaer,3 Fengji Xu,4 Yinhua Yu,4 Robert C. Bast,4 Jan Blaakaer,5 and Ian J. Jacobs6 OVX1 is a tumor marker that may be of value in the management of ovarian cancer and appears to be complementary to CA125 measurement (1 3 ). The results of a study by Woolas et al. (2 ) suggested that OVX1 is increased in the serum of a major proportion of patients with stage I ovarian cancer who have serum CA125 concentrations within the health-related reference interval. We, therefore, had planned to include OVX1 measurement in a prospective study of screening for ovarian cancer. Because the screening study involved sending blood samples by post to a central laboratory, we undertook this preliminary study to assess the stability of OVX1in the 2- to 3-day time period required for postal delivery. Clinical Chemistry 45, No. 5, 1999

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10 RESULTS: The results indicate that the OVX1 radioimmunoassay is highly dependent on sample handling. First, OVX1concentrations rose significantly if blood samples were allowed to clot at room temperature, but this effect was not seen if samples were kept at 4 C. Second, there were significant differences between serum and plasma samples. Serum samples once separated were stable on repetitive thawing and freezing.

11 Is tissue CA125 expression in epithelial ovarian adenocarcinoma heterogenic? MORTEN H. SPARHOLT, CLAUS K. HØGDALL, LOTTE NEDERGAARD, MEL C. J. HEERAN,IB J. CHRISTENSEN and ESTRID V. S. HØGDALL Purpose: To evaluate if heterogeneity of tissue cancer antigen 125 (CA125) expression is present in epithelial serous adenocarcinomas. Furthermore, to investigate whether there is a correlation between levels of CA125 tissue expression, serum level of CA125, stage, and grade. APMIS 2012, 121: 38 44

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13 RESULTS AND CONCLUSION: CA125 was expressed in varying degrees in tumor tissues from all patients. Mean tissue CA125 expression for each patient ranged from 36% to 98%. Intrapatient variations in tissue expression ranged from 10% to 90% point. No significant correlations between levels of CA125 tissue expression, serum level of CA125, stage, and grade were found. We found that the tissue expression of CA125 is heterogenic. Although most patients had a high mean expression, it covers a large intrapatient variation in expression. This suggests that if using CA125 as a tissue marker and anti-ca125 (oregovomab) as immunotherapy treatment in future studies, it will be necessary to take heterogeneity into consideration and examine a larger number of biopsies. Therefore, the study demonstrates the need for heterogeneity studies in future translational research.

14 Sample handling for mass spectrometric proteomic investigations of human urine Anette Lykke Petri1, 2, Claus Høgdall2, Ib Jarle Christensen3, Anja Hviid Simonsen4, Davy T Jampens4, Marja-Leena Hellmann4, Susanne Krüger Kjaer2, 5, Eric T. Fung4 and Estrid Høgdall5 Proteomics Clin. Appl. 2008, 2,

15 BAGGRUND OG FORMÅL: Because of its non-invasive sample collection method, human urine is an attractive biological material both for discovering biomarkers and for use in future screening trials for different diseases. Before urine can be used for these applications, standardized protocols for sample handling that optimize protein stability are required. In this explorative study, we examine the influence of different urine collection methods, storage temperatures, storage times, and repetitive freeze-thaw procedures on the protein profiles obtained by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS).

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18 CONCLUSION: suggestions for an optimal urine collection protocol for future urine analyses are: Urine samples should be collected using a single and consistently performed protocol. Urine samples should be collected at the same time of day. Urine should be stored in a refrigerator or on ice for a very limited period of time (max 6 h in the present study) until freezing. For consistency and clinical practice, handling of urine samples should be finalized with freezing at -80 C within same workday. Further freeze/thawing is possible up until eight rounds without any effect on the proteomic profiles.

19 CASE fra det rigtige liv GIST tumor 1988 gram præparatet fremfindes. To situationer..

20 Hvis der ikke selekteres. Billede af sekvens Analyseresultatet: Ingen mutationer i hverken c-kit eller PDGFRa

21 Hvis der selekteres. Analyseresultatet: c-kit exon 9 insertion c.1509_1510insgcctat (p.y503_y504insay (insertions - in frame) )

22 Tværfagligt samarbejde er forudsætningen for gode og pålidelige svar. Både præ-analytiske faktorer og hvordan vi behandler og opbevarer materiale er af stor betydning. Og nu til dagens første taler!