Telomerase serves essential functions in countering the end-replication problem and conferring immortal replicative potential upon cancer cells. Inhibition of telomerase has been shown in model systems to block tumor growth and has therefore been considered a promising anti-cancer therapeutic strategy. Successful implementation of anti-telomerase therapy has been hampered by an incomplete understanding of the telomerase enzyme and its mechanisms of action. Telomerase is regulated through a complex, yet essential assembly and trafficking pathway that culminates in recruitment of the enzyme to its physiological substrate, the chromosome end. Telomerase assembly requires specific assembly factors to produce an enzymatically active ribonucleoprotein comprising the telomerase reverse transcriptase (TERT), the telomerase RNA component (TERC) and dyskerin, a TERC-binding protein required for telomerase stability. In the final steps in the pathway, the telomerase protein TCAB1 enables telomerase accumulation within Cajal bodies, sites of ribonucleoprotein assembly and modification. After this step, telomerase is able to translocate to telomeres through a crucial, but incompletely understood process. This proposal studies the mechanisms by which telomerase associates with telomeres in human cancer cells. We dissect the association between telomerase and TPP1, a telomere-binding protein that serves roles both in protecting chromosome ends and in controlling telomerase access to telomeres. Telomerase can interact with the oligonucleotide/oligosaccharide binding fold (OB-fold) of TPP1 and we develop new assays that allow the interaction between telomerase and the OB-fold of TPP1 to be studied in vivo. We find that the OB-fold acts as a potent inhibitor of telomerase action at telomeres, inhibiting telomerase recruitment and causing rapid telomere shortening. Insights gained from understanding how telomerase is recruited to telomeres may enable the generation of targeted therapeutics that block the ability of telomerase to support immortal growth of tumors. To understand how telomerase is recruited to telomeres in human cancer cells, we will pursue the following specific aims: (1) To define the interaction between telomerase and TPP1 (2) To understand telomerase recruitment through real-time live cell imaging and (3) To determine the genetic requirement for TPP1-telomerase association in telomere maintenance.
Cancer is inherently linked to unlimited and dsyregulated proliferation of abnormal cells forming a tumor that can spread to other sites in the body, eventually killing its host. This behavior of cancer cells is inherently different from most cells within the body, but does share important commonalities with stem cells. Tumor cells and stem cells have a very large capacity to divide due in part to expression of the enzyme telomerase, which elongates telomeres that special caps that protect chromosome ends. Tumor cells and stem cells also share the ability to self- renew, that is, to divide in such a manner as to generat a daughter cell with all the capabilities of the original cell. Maintenance of telomeres by telomerase is a critical component of self- renewal in both stem cells and in cancer cells. However, how telomerase maintains telomeres remains poorly understood, which has limited our ability to develop therapeutics directed against this pathway. In this proposal, we will study how telomerase associates with specific telomere binding proteins to control telomere synthesis. Targeting this interaction may represent a major opportunity to inhibit this pathway.
|Batista, Luis F Z; Artandi, Steven E (2013) Understanding telomere diseases through analysis of patient-derived iPS cells. Curr Opin Genet Dev 23:526-33|
|Shkreli, Marina; Sarin, Kavita Y; Pech, Matthew F et al. (2012) Reversible cell-cycle entry in adult kidney podocytes through regulated control of telomerase and Wnt signaling. Nat Med 18:111-9|
|Zhong, Franklin; Savage, Sharon A; Shkreli, Marina et al. (2011) Disruption of telomerase trafficking by TCAB1 mutation causes dyskeratosis congenita. Genes Dev 25:11-6|
|Batista, Luis F Z; Pech, Matthew F; Zhong, Franklin L et al. (2011) Telomere shortening and loss of self-renewal in dyskeratosis congenita induced pluripotent stem cells. Nature 474:399-402|
|Park, Jae-Il; Venteicher, Andrew S; Hong, Ji Yeon et al. (2009) Telomerase modulates Wnt signalling by association with target gene chromatin. Nature 460:66-72|
|Venteicher, Andrew S; Abreu, Eladio B; Meng, Zhaojing et al. (2009) A human telomerase holoenzyme protein required for Cajal body localization and telomere synthesis. Science 323:644-8|
|Venteicher, Andrew S; Meng, Zhaojing; Mason, Philip J et al. (2008) Identification of ATPases pontin and reptin as telomerase components essential for holoenzyme assembly. Cell 132:945-57|
|Choi, Jinkuk; Southworth, Lucinda K; Sarin, Kavita Y et al. (2008) TERT promotes epithelial proliferation through transcriptional control of a Myc- and Wnt-related developmental program. PLoS Genet 4:e10|