Telomerase, Immortalization and Cancer Eric Bankaitis Cancer Bio 169 March 9, 2006

1. Telomerase, Immortalization and CancerEric BankaitisCancer Bio 169March 9, 2006 Fig.[9]

2. Kilian et al Telomeres: terminal regions of each chromosome. Functions: -Protect genome -Shield from DSB recognition. -Regulate the cell lifespan Fig.[9] Overview of Telomeres and Telomerase

3. Telomere Structure[12] -Repeating DNA sequences -Single stranded overhang -DNA binding proteins -Length varies from cell type to cell type, as do the DNA binding proteins. Fig.[9] Overview of Telomeres and Telomerase

4. Overview of Telomeres and Telomerase TELOMERE BINDING PROTEINS DNA Damage Control Stability and Structure Length Regulators Growth Factor Response Apoptosis Polymerase Co-factors RNA dependent Polymerase [1]

5. Telomerase Structure: RNA dependent DNA Polymerase Two Subunits: hTERT and hTR[1] Mutations eliminate activity figure[2] Overview of Telomeres and Telomerase

6. Telomerase Function: Maintain Telomeres! Senescence Crisis and the Hayflick Limit Strong Selection in Cancer [9] Overview of Telomeres and Telomerase

7. -Differentiated Tissue: Short Telomeres Telomerase INACTIVE -Germ cells, stem cells, cancer cells: Telomere Length Maintained Telomerase ACTIVE[1] Knockout Mice[4]: -Normal development -6-Generation Abonormalities: -CIN -Aging Why Cells Need Telomerase

8. Telomerase and Cancer • Statistics • Telomerase is activein 90% of Cancer[3].

9. Mechanisms in Cancer Development • Strong Selection for Immortalization in Cancer Progression • UPREGULATION OF TELOMERASE CATALYTIC SUBUNIT!!! • Multiple Hit Model[5] • Loss of Telomerase Repressors[6] • Oncogenic activation of Positive Regulators[7] • Enhancement of hTERT transcription[7]

10. Loss of Telomerase Repressors Differentiated Tissue: hTERT(-) Immortal Cells: hTERT(+) -Cell Fusion Experiments[5] -Repressor on Human Chromosome 3[5] “Changes in the telomeric binding proteins may regulate accessibility to the 3` overhang.”[5]

11. “E2F1 mRNA expression and hTERT mRNA expression were statistically significantly correlated in human glioblastoma cells.” “E2F1 expression increased hTERT promoter activity in these cells.” “We detected an interaction between E2F1 protein and hTERT promoter”[8] hTERT=> RB/E2F Pathway involved in some cells

12. “Several transcription factors, including oncogene products (e.g. c-Myc) and tumor supressor gene products are able to control hTERT transcription when overexpressed.[7]” [10] Oncogenic Activation of hTERT Expression

13. Viral Activation of hTERT “HPV E6 protein contributes to keratinocyte immortalization through trans-activation of hTERT gene transcription.”[7] “In some hepatocellular carcinomas, the hTERT gene is a non-random integration site for the HBV genome, which cis activates hTERT transcription.”[7]

14. Figure 5: TERT Expression in Colonic Tumorigenesis.  Source: Kolquist et al Treatment and Diagnostic Methods

15. Treatment and Diagnosis of Cancer[1] -Inhibitors of Telomerase: low side effects inhibitors already exist for many viral RT’s -Antisence RNAs -Inhibit Telomerase Access to the Telomere Potential Problem: ALT pathway.

16. References 1) Hahn. William C. American Society of Clinical Oncology. 2003. P2034-2043 2) Griffiths A., Gelbert W. Modern Genetic Analysis; Second Edition. 2003, W.H. Freeman and Co. p100. 3) Shay J and Bachetti S. European Journal of Cancer. Vol. 33, No. 5, p787-791, 1997. 4) Greider C.W., Blasco M.A., Cell. 1997 Oct 3, 91(1): 25-34. 5) Shay J, Wright W, Holt S. European Journal of Cancer. Vol. 33, No. 5, p761-766, 1997. 6) Barretr J, Oshimura M, European Journal of Cancer. Vol. 33, No. 5, pp710-715, 1997. 7) Horikawi L, Barret J. 2003 Jul’24(7):1167-76. Epub 2003 May 22. PMID: 12807729. 8) Alossa M, Gomez-Manzara C. 2005 Nov2;97(21): 1589-600. PMID: 16264179 9) Jerry Shay. University of Texas, Southwestern Medical Center, Dallas USA. 10) Lodish, Molecular Cell Biology: Firth Edition. W.H. Freeman and Company, New York. p 588.