Induced Pluripotent Stem Cell (iPSC) has opened up new approaches in disease modeling, personalized medicine, cell therapy, regenerative medicine and so on.

1. Modeling Human Disease
with iPSC
Tahsin Hasan Rahit
MDSC 641.04 Genomics (Fall 2018)
University of Calgary

2. 12 years of iPSC
Dr. Shinya Yamanaka Dr. Kazutoshi Takahashi
Adapted from Bob Argiropoulos Slide

3. Objectives
Understand the advantages of
using iPSC for disease modelling
Know how iPS cells are used in
disease modeling
Realize the possibilities of hiPSC
in future disease modeling

4. Outline
Why iPSC
Disease Modeling Example (Neural)
Challenges
Future Aspect

5. Why iPSC

6. Access to Critical Cells
Image: modified from bio-rad.com

7. Models Species
• Different pathophysiology and
genetic factors
iPSC: “Disease in vitro”
Having individual’s unique genotype
precisely (almost)
Image: Vera and Lorenz Development 142 18 (2015)

8. Disease Modelling
(Alzheimer’s Disease)

9. Alzheimer’s Disease
Image: Genetics Home Reference

10. Alzheimer’s Disease in-vitro
• Are they showing secretion of Aβ Peptite, Oligomers?
• Can we simulate toxicity and cell death?
Do they contain disease genotype & phenotype?

11. iPSC
Kondo, T. et al. Cell Stem Cell (2013).

12. iPSC
Kondo, T. et al. Cell Stem Cell (2013).
• Control: 3 healthy individual
• APP-E693Δ: 3 familial AD individual
• APP-V717L: 2 familial AD individual
• Sporadic AD: 2 unrelated AD individual

13. Cell Culture for Neural Differentiation
Kondo, T. et al. Cell Stem Cell (2013).
DMEM/F 12
KSR 5% N2 1x
Neurobasal
B27 (Vitamin A minus) 1x
Dorsomorphin 2μM
SB431542 10μM
GDNF/BDNF/NT3 10
day24 day65 day72
passage
day08
EB Floating Culture EB adherent Culture on Matrigel iPSC derived neuron adherent culture on Matrigel
(Culture neural cell from iPSC)

14. Neuron Cell
Kondo, T. et al. Cell Stem Cell (2013).
cortical neuron (subtype SATB2 & TBR1)

15. Aβ Oligomers
Kondo, T. et al. Cell Stem Cell (2013).

16. Stress & Cell Death
Kondo, T. et al. Cell Stem Cell (2013).

17. Alzheimer’s Disease in-vitro
• Are they showing secretion of Aβ Peptite, Oligomers?
• Can we induce toxicity and simulate cell death?
Do they contain disease genotype & phenotype?

18. Challenges
• Variations in differentiation efficiency [Miura, K. et al. (2009), Koyanagi-Aoi, M. et al. (2013)]
• Choose truly pluripotent iPSC [Takahashi, K. & Yamanaka (2016)]
• Line-to-line variations in the maturation of differentiated cells
• Culture condition [Inoue, H. et al. (2014)]
• microRNA switch [Miki, K. et al. (2015)]

19. Not only Monogenetic

20. 3D & Beyond
Image: Chun Liu et al. Development (2018)

21. Chun Liu et al. Development (2018)
3D iPSCs Disease Modelling Studies

22. Conclusion
• Sporadic disease study
• Simulation without limitation
• Debugger (borrowed the term from CS)

23. References
1. Ardhanareeswaran, K., Mariani, J., Coppola, G., Abyzov, A. & Vaccarino, F. M. Human induced pluripotent stem cells for modelling neurodevelopmental disorders. Nat. Rev.
Neurol. 13, 265–278 (2017).
2. Chen, Y.-W. et al. A three-dimensional model of human lung development and disease from pluripotent stem cells. Nat. Cell Biol. 19, 542–549 (2017).
3. Dutta, D., Heo, I. & Clevers, H. Disease Modeling in Stem Cell-Derived 3D Organoid Systems. Trends Mol. Med. 23, 393–410 (2017).
4. Ebert, A. D., Liang, P. & Wu, J. C. Induced pluripotent stem cells as a disease modeling and drug screening platform. J. Cardiovasc. Pharmacol. 60, 408–16 (2012).
5. Kondo, T. et al. Modeling Alzheimer’s disease with iPSCs reveals stress phenotypes associated with intracellular Aβ and differential drug responsiveness. Cell Stem Cell 12,
487–96 (2013).
6. Li, L., Chao, J. & Shi, Y. Modeling neurological diseases using iPSC-derived neural cells. Cell Tissue Res. 371, 143–151 (2018).
7. Liu, C., Oikonomopoulos, A., Sayed, N. & Wu, J. C. Modeling human diseases with induced pluripotent stem cells: from 2D to 3D and beyond. Development 145, dev156166
(2018).
8. Shaheen, N., Shiti, A. & Gepstein, L. Pluripotent Stem Cell-Based Platforms in Cardiac Disease Modeling and Drug Testing. Clin. Pharmacol. Ther. 102, 203–208 (2017).
9. Shi, Y., Inoue, H., Wu, J. C. & Yamanaka, S. Induced pluripotent stem cell technology: a decade of progress. Nat. Rev. Drug Discov. 16, 115–130 (2017).
10. Siller, R., Greenhough, S., Park, I.-H. & Sullivan, G. J. Modelling human disease with pluripotent stem cells. Curr. Gene Ther. 13, 99–110 (2013).
11. Takahashi, K. et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131, 861–72 (2007).
12. Takahashi, K. & Yamanaka, S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–76 (2006).
13. Workman, M. J. et al. Engineered human pluripotent-stem-cell-derived intestinal tissues with a functional enteric nervous system. Nat. Med. 23, 49–59 (2017).