1. High expression of Pyruvate Dehydrogenase Kinase in
Cholangiocarcinoma patients
Miss Surangkana Sanmai
Faculty of Associated Medical Sciences, Khon Kaen University

2. Cholangiocarcinoma Major risk factor Cholangiocarcinoma (CCA)
Solid tumor which arise of biliary epithelial cell.
Arise from chronic inflammation
intrahepatic cholangiocarcinoma
perihilar cholangiocarcinoma
distal cholangiocarcinoma
Major risk factor
Sripa, 2012

3. Diagnosis Diagnosis The low specificity Computed tomography scan
Magnetic resonance imaging
Biomarkers : CEA, CA19-9, SBG, MUC5AC etc.
The low specificity

4. Drug and drug combination
Therapy of CCA
Surgical resection (15-35%)
Radiation
Chemotherapy; Gemcitabine, 5-FU, Gemcitabine plus cisplatin, etc.
No.
Drug and drug combination
mechanisms
Overall response
(%)
References 1
5-FU alone &
combinations
nucleoside analog
10-40%
Falkson et al.1984, Choi et al. 2000,
Gebbia et al. 1996, Patt et al. 1996,
Raderer et al. 1999, Taieb et al. 2002,
Ellis et al. 1995, Lee et al. 2004,
Ducreux et al. 1998, 2
Gemcitabine alone & combinations
thymidylate synthase inhibitor
14-50%
Arroyo et al. 2001, Gebbia et al. 2001,
Raderer et al. 1999, Penz et al. 2001,
Gebbia et al. 2001, Carraro et al. 2001,
Andre et al. 2004, Bhargava et al. 2003,
Thongprasert et al. 2005, Knox et al. 2003
Sookprasert, 2012
Discovering new targeted drugs
or new strategy to treat CCA
Chemotherapy resistance & their side effects

5. Mitochondria and cancer
The mitochondria roles;
Redox balancing
The vital regulation of several cell
survival/death pathways.
Bioenergetics
Animal_mitochondrion_diagram_en_(edit).svg/400px-Animal_mito
chondrion_diagram_en_(edit).svg.png

6. Mitochondria and cancer
The regulation of glucose metabolism in cancer cells. When glucose enters the cell through a glucose transporter, it is phosphorylated by HK to glucose-6-phosphate, which is further metabolized by glycolysis to pyruvate in the cytosol. Under aerobic conditions, normal cells use pyruvate dehydrogenase (PDH) to convert most pyruvate to acetyl-CoA. The acetyl-CoA is then oxidized via the TCA cycle, providing sources of ATP synthesis. In contrast, the metabolic pathways of glucose utilization in cancer are changed from ATP generation by oxidative phosphorylation to ATP generation through glycolysis. Also, for cell proliferation to occur, cancer cells require the synthesis of new macromolecules (for example, nucleic acids, lipids, proteins). Key enzymes that may be promising targets for cancer therapy are highlighted in red. TCA enzymes that are known to be mutated in cancer are shown in purple: IDH2, SDH, and FH
The-regulation-of-glucose-metabolism-in-cancer-cells-When-glucose-enters-the-cell.png

7. Glucose metabolism-involved protein selection
The glucose metabolism-involved proteins in mitochondria of CCA
Table 1. The glucose metabolism-involved proteins in mitochondria of CCA (Chua-On et al., 2016)
No.
Protein name
Fold change
Papillary CCA
Non-papillary CCA 1
Pyruvate dehydrogenase kinase 3 13 14 2
Cytochrome c oxidase assembly protein 3 homolog 7 4 3
Cytochrome c oxidase subunit 5A
ATP synthase subunit epsilon 6 5
Cytochrome c oxidase subunit Vb -2
Complex I intermediate-associated protein 30
Succinate dehydrogenase 8
Pyruvate dehydrogenase kinase 2 9
Acetyl-CoA carboxylase 2 10
NADH dehydrogenase 11
ATP synthase subunit alpha, mitochondrial isoform -1 12
Cytochrome b-c1 complex subunit 1 -3
ATP synthase mitochondrial F1 complex
Isocitrate dehydrogenase 15
Cytochrome oxidase subunit I -5
and 16 proteins were included. Among 16 proteins, pyruvate
dehydrogenase kinase 3 (PDK) shows the highest fold change. Therefore, the inhibition of PDK may contribute a new strategy to CCA treatment.

8. Heat map

9. Pyruvate dehydrogenase kinases
The PDK is a Ser/Thr kinase
that inactivates mitochondrial
pyruvate dehydrogenase (PDH)
by site-specific phosphorylation.
PDKs
PDK1
PDK2
PDK3
PDK4
Size (kDa) 47 46
Tissue
Heart, colon cancer
All tissues
Testis, colon cancer
Skeletal muscle
PDH
Objective: To study the expression of PDKs expression in
CCA patients tissues
Suebwai, 2015

10. Results 1. The screening the expression of all PDK isoforms in CCA patient
tissues using immunohistochemistry
PDK1
Adjacent
CCA
Adjacent
CCA
Adjacent
CCA

11. PDK2
Adjacent
CCA
Adjacent
CCA
Adjacent
CCA

12. PDK3
Adjacent
CCA
Adjacent
CCA
Adjacent
CCA

13. PDK4
Adjacent
CCA
Adjacent
CCA
Adjacent
CCA

14. PDK4
Adjacent
CCA
Adjacent
CCA
Adjacent
CCA

15. Results: Comparison of PDKs levels in adjacent and cancerous tissues
Figure 2 Mann-Whitney U test showed that the overexpression of PDK1, PDK2 and PDK3
was significantly found in CCA tissues when compared to its adjacent non-cancerous tissues.
(p < ). (NS=Not seen)

16. Conclusions
The specification of PDKs pattern in CCA may provide a good tumor marker for CCA. Futhermore, PDK1 and PDK3 may be considered as potential prognostic markers for CCA patients due to their tumor progression involvement.
There are many glucose metabolism-involved proteins out of PDK that can be studied their roles and considered as a novel tumor markers.
Despite of tumor marker, PDK could be study in other roles such as drug target or prognostic marker or in other cancers.

17. Acknowledgement Dr. Sittiruk Roytrakul
Assoc. Prof. Dr. Siriporn Proungvitaya
Asst. Prof. Dr. Tanakorn Proungvitaya
Prof. Dr. Temduang Limpaiboon
Asst. Prof. Dr. Wunchana Seubwai
Doungdean Tummanatsakun
Daraporn Chua-on
The Research Fund for Supporting Lecturer to Admit High Potential Student to Study and Research on His Expert Program Year 2014, Graduate School, Khon Kaen University