1 / 15

Barbara DEWAELE, PhD Department of Human Genetics KU Leuven - UZ Leuven Leuven, Belgium

Identification of a novel , recurrent MBTD1-CXorf67 fusion in low grade endometrial stromal sarcoma. Barbara DEWAELE, PhD Department of Human Genetics KU Leuven - UZ Leuven Leuven, Belgium. November 16 th 2012 |CTOS 17th Annual Meeting | Hilton Prague in Prague, Czech Republic.

kalea
Download Presentation

Barbara DEWAELE, PhD Department of Human Genetics KU Leuven - UZ Leuven Leuven, Belgium

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  1. Identification of a novel, recurrentMBTD1-CXorf67fusion in low gradeendometrial stromal sarcoma Barbara DEWAELE, PhD Department of Human Genetics KU Leuven - UZ Leuven Leuven, Belgium November 16th 2012 |CTOS 17th Annual Meeting | Hilton Prague in Prague, CzechRepublic

  2. Endometrial stromal sarcomas (ESS) are rare mesenchymal tumors Non-pleomorphic Pleomorphic • Endometrialstromal tumors (EST) tissue architecture: • proliferativephaseendometrialstroma • EST represent a broad spectrum of tumors withvaryingdegrees of malignancy: • CurrentWHO classification: • BENIGN LOW-MALIGNANT MALIGNANT • ESN ESS UES ESS UES High-grade Low-grade JAZF1-SUZ12 JAZF1-PHF1 EPC1-PHF1 … YWHAE- FAM22 Complex karyotype Nuclearsize/irregularity in nuclear contour Mitoticactivity Tumor necrosis High mitoticactivity Tumor necrosis GoodprognosisIntermediateprognosis Poorprognosis AdaptedfromLee et al., Am. J. Surg. Pathol 2012: 36: 641-653.

  3. Aim: identification of novelfusions in ESS • Selection of two cases with t(X;17) byconventionalkaryotyping • Clinicallyandmorphologically: low grade ESS • Lackknownpathognomonic gene fusions of low grade ESS

  4. Clinicohistologicaland Immunohistochemical features of two t(X;17) ESS cases CD10 ER CD99

  5. Conventionalkaryotypingreveals the t(X;17) in two ESS cases dicentricchr. Case 2 Case 1 2 3 4 5 1 2 3 4 5 1 X X 6 7 8 9 10 11 12 6 7 8 9 10 11 12 13 14 15 16 17 18 13 14 15 16 17 18 19 20 21 22 19 20 21 22 45,X,t(X;17)(p11.2;q23),dic(4;22)(p15;q22) 46,X,t(X;17)(p11.2;q23)

  6. Conventionalkaryotypingreveals the t(X;17) in two ESS cases dicentricchr. Case 1 Case 2 1 2 3 4 5 1 2 3 4 5 X X 6 7 8 9 10 11 12 6 7 8 9 10 11 12 13 14 15 16 17 18 13 14 15 16 17 18 19 20 21 22 19 20 21 22 X derX 17 der17 X derX 17 der17

  7. Conventionalkaryotypingreveals the t(X;17) in two ESS cases andarray CGH indicatesthat the t(X;17) is balanced dicentricchr. Case 1 Case 2 2 3 4 5 1 2 3 4 5 X X 6 7 8 9 10 11 12 6 7 8 9 10 11 12 13 14 15 16 17 18 13 14 15 16 17 18 19 20 21 22 19 20 21 22 45,X,t(X;17)(p11.2;q23),dic(4;22)(p15;q22) 46,X,t(X;17)(p11.2;q23) Agilent 180K Case 1 Case 2 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 X Y

  8. Using RNA-seq, the MBTD1-CXorf67fusion was identified in the two ESS cases with the t(X;17) 17q21.33 MBTD1 (-) Chr. 17 • 2 ESS cases with t(X;17) • Paired-end RNA seq on Illumina platform • Case 1: 1,21 x 108 paired-end readsmappedto hg19 • Case 2: 6,29 x 107 • Analysis withArrayStudio and deFuse: • Case 1: 25 distinct and high-quality paired-end reads • Case 2: 2 link exon 16 of MBTD1 to exon 1 of CXorf67 q25.3 p13.3 Xp11.22 CXorf67 (+) Chr. X p22.33 q28 CXorf67 MBTD1 cDNA 1 16 1 Case 1 Reads 0 CXorf67 MBTD1 60 Case 2 0 16 1 3 5 7 13 17 1 84 kb 2,3 kb 40 Centromere Telomere

  9. Using RNA-seq, the MBTD1-CXorf67fusion was identified in the two ESS cases with the t(X;17) 17q21.33 MBTD1 (-) Chr. 17 • 2 ESS cases with t(X;17) • Paired-end RNA seq on Illumina platform • Case 1: 1,21 x 108 paired-end readsmappedto hg19 • Case 2: 6,29 x 107 • Analysis withArrayStudio and deFuse: • Case 1: 25 distinct and high-quality paired-end reads • Case 2: 2 link exon 16 of MBTD1 to exon 1 of CXorf67 q25.3 p13.3 Xp11.22 CXorf67 (+) Chr. X p22.33 q28 CXorf67 MBTD1 cDNA 1 16 1 Case 1 Reads CXorf67 MBTD1 60 Case 2 16 1 3 5 7 13 17 1 0 84 kb 2,3 kb 40 Centromere Telomere 0

  10. The MBTD1-CXorf67fusionupregulates the expression of the 3’ end of CXorf67 exon 1 control control control control MBTD1_exon 17 CXorf67 3’ end YWHAE-FAM22 YWHAE-FAM22 MBTD1_exon 15_16 CXorf67 5’ end JAZF1-SUZ12 JAZF1-SUZ12 MBTD1-CXorf67 MBTD1-CXorf67 Case 1 Case 1 MBTD1-CXorf67 MBTD1-CXorf67 Case 2 Case 2 Case 1 Reads CXorf67 MBTD1 60 Case 2 16 1 3 5 7 13 17 1 1 0 84 kb 2,3 kb 40 Centromere Telomere 0

  11. Using nested RT-PCR followedbySangersequencing, the MBTD1-CXorf67fusion was validated 141 245 351 456 1 FCS MBT1 MBT2 MBT3 MBT4 45 80 253 350 464 560 RNA Seq: control case 1 control case 2 200bp CXorf67 MBTD1 cDNA Case 2 503 MBTD1 exon 16 CXorf67 exon 1 Ser-rich ladder Case 1 K K S Q Q G N P A R L S P E Y H K 1 16 1 protein

  12. Expressionprofilingwithhierarchical clustering demonstrates clustering of JAZF1-SUZ12andMBTD1-CXorf67 low grade ESS 70 genes YWHAE-FAM22A JAZF1-SUZ12 JAZF1-SUZ12 YWHAE-FAM22A MBTD1-CXorf67 JAZF1-SUZ12 JAZF1-SUZ12 YWHAE-FAM22A MBTD1-CXorf67 ESS high ESS low UES

  13. One additional low grade ESS case positive for the MBTD1-CXorf67 fusion was identified using a specific FISH assay • Specific FISH assay todetect the t(X;17) translocation • 14 ESS (4 low grade ESS, 5 high grade ESS and 5 UES) • negative for JAZF1andYWHAErearrangements • paraffine tissues • identification of 1 additional case positive for MBTD1-CXorf67fusion

  14. Conclusions: • In thisstudy: • new gene fusion: MBTD1-CXorf67 • yet another cytogenetically distinct subgroup of ESS • based on clinico-histopathologic features and expression profiling: cluster together with • JAZF1-SUZ12 as low grade ESS • offers the opportunity to shed light on the functions of two poorly characterized genes • MBTD1 belongs to the Polycomb group (PcG) of proteins • CXorf67 : a yet uncharacterized protein • Future: • decipher downstream consequences of the fusion • evaluate the respective contributions of the MBTD1 and CXorf67 portions of the fusion • underlying oncogenic processes

  15. Acknowledgements: Department of Gynaecology: Frederic Amant InstitutBergonie, Bordeaux, France: Sabrina Croce The MSCMC and Institute of Oncology, Warsaw, Poland: Joanna Przybyl • Department of Human Genetics: Barbara Dewaele Maria Debiec-Rychter Jan Cools Peter Vandenberghe Vanessa Vanspauwen Quattrone Anna Julio Finalet Ferreiro Geerdens Ellen GianfeliciValentina KalenderZeynep • Departmentof Pathology: Raf Sciot Philippe Moerman • Department of Oncology: AgnieszkaWozniak

More Related