Sidney H. Ingbar Lecture, ATA 10/17/2013: PAX8-PPAR γ Fusion Protein in thyroid carcinoma Ronald J. Koenig, MD, PhD Division of Metabolism, Endocrinology & Diabetes University of Michigan PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Learning Objectives 1. Know what the PAX8-PPAR Fusion Protein is. 2. Know what types of thyroid cancer are associated with the PAX8-PPAR Fusion Protein. 3. Know about a clinical trial specifically designed for patients with PAX8-PPAR Fusion Protein thyroid cancer. 4. Know the normal function of PPAR . 5 Know about the Wnt pathway. (No disclosures) PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Outline of today’s presentation Background PPFP carcinoma patient samples Mouse model of PPFP thyroid carcinoma PPFP cell lines Summary and conclusions PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Gene mutations underlie thyroid carcinoma Vu-Phan D & Koenig RJ. Mol. Cell. Endocrinol., in press (review) PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
PPFP is generated by a chromosomal translocation Paired Box Gene 8 ( PAX8 ): Chr 2q13 1 2 3 4 5 6 7 8 9 10 11 12 Translocation (2;3)(q13;p25) 1 2 3 4 5 6 Peroxisome Proliferator-Activated Receptor Gamma ( PPARG ): Chr 3p25 DBD DBD LBD Fusion junction PPAR PAX8 PAX8-PPAR Fusion Protein (PPFP) PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Functions of PAX8 and PPAR . PAX8 is a transcription factor that is essential for thyroid development and that drives the expression of many thyroid-specific genes. PPAR is the master regulator of adipogenesis , among other functions. PPAR is expressed at very low levels in normal thyroid and has no known function in that organ. PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
PAX8-PPAR has dominant negative activity over PPAR (transfections in U2OS osteosarcoma cells) Kroll TG, et al . Science 289:1357-60, 2000 PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
PPAR , a putative tumor suppressor Several mouse models of cancer are exacerbated by a single allele deletion of Pparg , including a model of thyroid cancer that is unrelated to PPFP (Kato, et al . Oncogene 25:2736, 2006). Hypothesis : PPFP is oncogenic because it inhibits the tumor suppressor activity of PPAR . PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Outline of today’s presentation Background PPFP carcinoma patient samples Mouse model of PPFP thyroid carcinoma PPFP cell lines Summary and conclusions PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Gene expression signature of PPFP follicular thyroid cancers Thomas Giordano, MD, PhD FC(+) FC(-) NT FA OA OC PC AC MC Giordano TG, et al . Clin Cancer Res. 12:1983-93, 2006 PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Gene expression signature of PPFP follicular thyroid cancers. FC(+) FC(-) NT FA OA OC PC AC MC AQP7 ANGPTL4 Giordano TG, et al . Clin Cancer Res. 12:1983-93, 2006. PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
PPFP has PPAR -like activity on the AQP7 promoter. AQP7-Luc -Cz +Cz 20 6 JEG-3 N2a 5 16 4 12 3 8 2 4 1 Fold Activity Luc/Rluc 0 0 6 60 FRTL-5 Dog thyrocytes 5 50 4 40 3 30 2 20 1 10 0 0 PPAR PPAR pCDNA PAX8 PPFP pCDNA PAX8 PPFP Giordano TG, et al . Clin Cancer Res. 12:1983-93, 2006. PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Outline of today’s presentation Background PPFP carcinoma patient samples Mouse model of PPFP thyroid carcinoma PPFP cell lines Summary and conclusions PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Thyroidal expression of PPFP in transgenic mice Melissa Dobson, PhD Ericka Diallo-Krou CAG promoter PPFP eGFP intron Lox P Lox P X TPO promoter Cre Dobson ME, et al . Endocrinology. 152:4455-65, 2011 PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Thyroidal expression of PPFP in transgenic mice CAG promoter PPFP eGFP intron Lox P Lox P X TPO promoter Cre 40 Wild type PPFP;Cre PPFP/PPAR 30 20 10 Hyperplasia, 0 PPFP;Cre No neoplasia mean±SD, n=14 Dobson ME, et al . Endocrinology. 152:4455-65, 2011 PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
FTC pAKT PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
FTC pAKT PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
PPFP carcinomas have increased phosphoAKT pAKT(S473) Immunohistochemistry Normal human thyroid PPFP cancer Diallo-Krou E, et al . Endocrinology. 150:5181-90, 2009 PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
PPFP carcinomas have increased phosphoAKT pAKT(S473) Immunohistochemistry Normal human thyroid PPFP cancer Combined PPFP expression with thyroid- Mouse Thyroid AKT immunoblot specific Pten deletion: PPFP;PtenFF;Cre Control mice Pten deletion: PtenFF;Cre. pAKT(T308) pAKT(S473) Total AKT -actin PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Mouse thyroid glands Wild type PtenFF;Cre PPFP;PtenFF;Cre Dobson ME, et al . Endocrinology. 152:4455-65, 2011 PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
PPFP;PtenFF;Cre mice develop metastatic thyroid carcinoma Tracheal Invasion Vascular Invasion H&E Myc (PPFP) TTF-1 Lung Metastases H&E Myc (PPFP) TTF-1 PAX8 (PtenFF;Cre mice have benign thyroid hyperplasia with no neoplasia). Dobson ME, et al . Endocrinology. 152:4455-65, 2011 PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Pioglitazone therapy of PPFP;PtenFF;Cre thyroid cancer Thyroid size by ultrasound PPFP;PtenFF;Cre PtenFF;Cre Control Diet Pioglitazone 120- 110- Thyroid Area (mm 2 ) 100- 90- 18- 80- 16- 70- 14- 60- 12- 50- 10- 40- 8- 30- 6- 20- 4- 10- 2- 0- Start End 0- Start End WT thyroid area 1.9±0.5 mm 2 Dobson ME, et al . Endocrinology. 152:4455-65, 2011 PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Pioglitazone therapy of PPFP;PtenFF;Cre thyroid cancer. Thyroid size by ultrasound Pioglitazone prevents local invasion and PPFP;PtenFF;Cre PtenFF;Cre metastases in PPFP;PtenFF;Cre mice Control Diet Pioglitazone 120- Local Vascular Distant 110- N invasion invasion mets Thyroid Area (mm 2 ) 100- 90- 18- Control diet 12 11 5 7 80- 16- 70- 14- Pioglitazone 10 1 0 0 60- 12- 50- 10- P 0.0002 0.03 0.005 40- 8- 30- 6- (Neither invasion nor metastases were 20- 4- 10- 2- found in any PtenFF;Cre mice) 0- Start End 0- Start End WT thyroid area 1.9±0.5 mm 2 Dobson ME, et al . Endocrinology. 152:4455-65, 2011. PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Adipogenic PPAR -like response to pioglitazone in PPFP;PtenFF;Cre thyroids Control Diet Pioglitazone PtenFF;Cre PtenFF;Cre PPFP; Lipid droplets 100 Dobson ME, et al . Endocrinology. 152:4455-65, 2011 PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
Adipogenic PPAR -like response to pioglitazone in PPFP;PtenFF;Cre thyroids Control Diet Pioglitazone PPFP;PtenFF;Cre +Pio PtenFF;Cre Oil Red O Perilipin-1 PtenFF;Cre PPFP; Lipid droplets 100 Dobson ME, et al . Endocrinology. 152:4455-65, 2011 PRESENTATION FROM THE 83rd ANNUAL MEETING OF THE AMERICAN THYROID ASSOCIATION, OCTOBER 16-20, 2013 (Ronald J. Koenig)
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