Telomerase Inhibitor: Past, Present and Future

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Telomerase Inhibitor: Past, Present and Future Hematology Fellow Conference Janghee Woo, MD PhD University of Washington H. Joachim Deeg, MD Fred Hutchinson Cancer Research Center

Telomere & End replication problem http://www.nobelprize.org

Telomere and Telomerase Telomeres are repeated DNA sequences, by Elizabeth Blackburn, 1978 Telomeric DNA from Tetrahymena protected yeast cells from degradation and senescence during cell doubling, by Jack Szostak and Elizabeth Blackburn, 1982 Telomerase, a riboprotein enzyme, discovered to extend telomere DNA, preventing senescence, by Carol Greider and Elizabeth Blackburn, 1985 Telomerase uses a specific RNA template to specify the addition of telomeric repeats via a reverse transcriptase mechanism, by Carol Greider and Elizabeth Blackburn, 1989 Nobel Prize in Physiology or Medicine 2009 Elizabeth H. Blackburn, Carol W. Greider and Jack W. Szostak "for the discovery of how chromosomes are protected by telomeres and the enzyme telomerase"

Telomerase and Predicted biological effects Telomerase expression is essential for the proliferation of most cancer cells, but the enzyme is inactive in the majority of normal human tissues. White et al. Trends in Biotechnology, 2001

Imetelstat: First in Class Telmoerase Inhibitor Herbert et al., Oncogene, 2005

Imetelstat inhibits telomerase activity, htert, and telomere length in vitro Shammas et al., Leukemia, 2008

Telomerase Puterbation Trials in Cancer Denmark s Pharmexa prematurely halted in 2011, a phase 3 trial that was testing GV1001, a vaccine designed to prime the immune system to recognize telomerase, in 520 people with pancreatic cancer. Geron discontinued a phase 2 study of imetelstat in 166 patients with breast cancer in 2012 no survival benefit in interim analysis. A randomized phase 2 study of imetelstat as maintenance therapy for advanced non-small-cell lung cancer in 116 patients: no survival benefit (Chiappori et al., Ann Oncol. 2015) Thrombocytopenia was noted as an adverse effect in earlier failed phase 2 trials of imetelstat in people with breast and lung cancer.

Dyskeratosis congenita (DC) X-linked DC I. DKC1 gene, ribosomal RNA processing and telomerase function by stabilization of TERC RNA Autosomal dominant DC Heterozygous mutations in I. TERT gene, telomerase reverse transcriptase II. III. TERC gene, an essential RNA subunit of telomerase TINF2, a component of the shelterin telomere-associated protein complex Disease manifestations may become progressively more severe and develop at an earlier age over generations, disease anticipation.

Telomere in MPN High telomerase activity in granulocytes from clonal polycythemia vera and essential thrombocythemia. (Ferraris et al., Blood, 2005) Anna Maria Ferraris et al. Blood, 2005

Telomere in MPN High telomerase activity in granulocytes from clonal polycythemia vera and essential thrombocythemia. (Ferraris et al., Blood, 2005) Telomere length is reduced in JAK2V617F- positive and negative myeloproliferative neoplasms. (Bernard et al., Leukemia, 2009)

Terc / LSC engraftment progressively decreased, demonstrating that Terc / LSCs are severely limited in their ability to expand Bruedugam et al., Cell Stem Cell, 2014

Differential effects of Imetelstat on hematopoietic progenitor cells from MF and ET patients Imetelstat selectively inhibits the proliferation of MF hematopoietic progenitor cells by promoting apoptosis and depleting malignant MF HPCs. Imetelstat selectively affects malignant megakaryopoiesis in Myeloproliferative Neoplasms (MPN) Wang et. al. and Iancu-Rubin et. al, 2014 ASH

Outstanding questions Who will respond? : splicing mutations (CR in 3 out of 8 patients with mutations vs 1 of 25 patients without mutations) ASXL1 mutation (RR 32%, 7 of 22 patients without mutations vs vs 0 of 11 patients with an ASXL1 mutation (P = 0.07) Is telomere a real target? : no consistent pattern of drug effect on telomere length

No consistent pattern of drug effect on telomere length Baseline and post-treatment evaluation of telomere length among 5 patients with complete or partial remissions.

Outstanding questions Who will respond? : splicing mutations (CR in 3 out of 8 patients with mutations vs 1 of 25 patients without mutations) ASXL1 mutation (RR 32%, 7 of 22 patients without mutations vs vs 0 of 11 patients with an ASXL1 mutation (P = 0.07) Is telomere a real target? : no consistent pattern of drug effect on telomere length Predictive markers and Mechanism of action/resistance

Hutch Protocol #9435 A Randomized, Single-Blind, Multicenter Phase 2 Study to Evaluate the Activity of 2 Dose Levels of Imetelstat in Subjects with Intermediate-2 or High-Risk Myelofibrosis (MF) Relapsed/Refractory to Janus Kinase (JAK) Inhibitor

Inclusion Criteria

Primary endpoints

Secondary endpoints

Exploratory endpoints

Mr. P, 66 y/o male ET diagnosed in 1992, progressed Post-ET MF in 2005, CALR mutation (52bp deletion) Hydrea: diagnosis 2008 Jakafi: July 2013 Pegylated Interferon: Aug 2015 Portal hypertension, intractable ascites and variceal bleeding s/p TIPS in 2013, resolved. Non-transplantation candidate Refractory to Jakafi 20mg bid, increasing fatigue, splenomegaly symptoms, RBC transfusion dependent Spleen: 26.5 cm WBC 15.1, Hgb/Hct 5.2/16, Plt 138, CD34 in marrow 3.5 %, normal cytogenetics, peripheral blood blast 1.21% His DIPSS score is a 5 with a breakdown as follows: Age >65 = 1. Hemaglobin <10 = 2, Luekocytes >25K = 0, Circulating Blasts >1% = 1, Constitutaional Symptoms = 1

Off Jakafi/On Pred

Off Jakafi/On Pred 1 st Imetelstat 2 nd Imetelstat Symptomatic improvement

1978 1982 1985 1989 2005 2013-4 2015 Telomeres Telomere function Telomerase htr/htert Imetelstat Phase I ET/MF Phase II PAST: From bench to bedside: Telomerase inhibitor, imetelstat PAST: Proof of Concept phase I trials in patients with ET and MF Molecular response Reversal of marrow fibrosis PRESENT: Phase II trial in patients with MF, refractory to or non-tolerating JAK inhibitor, (non-transplant candidate) FUTURE: Biology of disease, Mechanism of action/resistance of imetelstat

Acknowledgements H. Joachim Deeg Bart Scott Elizabeth A. Stohr Sioban Keel

Q & A

Baseline telomere length comparisons Baseline telomere length comparisons in patients with or without complete/partial remissions following treatment with imetelstat

Alternative lengthening of telomeres (ALT) O Sullivan et al., Trends in Cell Biology, 2014