1. CFCF Patient Education Michael Y. Choi, MD September 12, 2009 Using gastrin producing G-cell and its essential gene Nkx6.3 as a model to study gastrinoma

2. Mucous CellParietal Cell Gastroendocrine CellChief Cell Gastric Lineages Stomach Stem Cell

3. Nkx6.3 Expression

4. Decreased G-cell Number in the Nkx6.3-null Stomach

5. Abrogated Gastrin mRNA Expression in Nkx6.3 Stomach

6. Physiology Review of G- and D-cells

7. Co-localization of Nkx6.3 and Gastrin in the Stomach

8. Serum GastrinGastric pH Wild type (N = 7) Knockout (N = 7) Wild type (N = 16) Knockout (N = 7) Mean87.1 ng/L37.7 ng/L3.95.5 SEM11.0 ng.L10.8 ng/L0.3 P-value0.00750.0032 Circulating gastrin levels and stomach luminal pH in Nkx6.3-/- and control animals

9. Mutant mice strains that have altered G-cell numbers Gastric Endocrine Cell Type Number in KO Mutant KO StrainRef.GastrinSomatostatinSerotoninGhrelin Pdx1 -/-Larsson et al.DownUnchangedUp Pax6 -/-Larsson et al.Down Unchanged Ngn3 -/-Jenny et al.Down Unchanged Lee et al.Down CCK-B/Gastrin Receptor -/- Langhans et al.UpDown Gastrin -/-Zavros et al.DownUnchanged Nkx6.3 -/-DownUpUnchanged

10. Relative expression levels of Pdx1, Pax6, and Ngn3 mRNA are unchanged in adult Nkx6.3-null stomach

11. Nkx6.3 mRNA levels are unchanged in neonatal Ngn3-null antrum

12. Modified from Hochedlinger, K. et al. Development 2009 The developmental potential and epigenetic states of cells at different stages of development PancreasStomach

13. Generating induced pluripotent stem cells by direct reprogramming Showed that iPS cells can be generated from mouse fibroblasts by retrovirus-mediated introduction of four transcription factors: - Sox2, Oct4, Klf4, and c-Myc

14. Yamanaka: Convert tail fibroblasts into pluripotent cells resembling embryonic stem cells by expressing 4 genes: Oct4 Sox2 Klf4 cMyc “An Astonishing Experiment” – Ian Wilmut, Cloning and Stem Cells

15. Generating insulin secreting  cells by direct reprogramming Expressed 3 genes Ngn3, Pdx1, Mafa to generate  -cells from exocrine cells

16. Cell Reprogamming: what’s the limit?  -Cells Excocrine Cells Ngn3, Pdx1, Mafa ?Cancer Cells ? Stem CellsFibroblasts Sox2, Oct4, Klf4, Myc G-Cells (or any cell type) Non G-Cells Nkx6.3, Pdx1, Ngn3, Pax6, etc.?

17. Nkx6.3 upregulates Gastrin in 293 cells

18. Gastrinoma Endoscopic view and a nest of tumor cells in lamina propria mucosae

19. Acknowledgement Ramesh Shivdasani Maina Lepourcelet Anthony Romer Qiufu Ma Paul Grey Melissa Wu Andrew Leiter Yang Wang Susumu Ito Byeong-Moo Kim Daniel Podolsky