Antisense STAT3 inhibitor decreases viability of myelodysplastic and leukemic stem cells
Aditi Shastri, Gaurav Choudhary, Margarida Teixeira, Shanisha Gordon-Mitchell, Nandini Ramachandra, Lumie Bernard, Sanchari Bhattacharyya, Robert Lopez, Kith Pradhan, Orsolya Giricz, Goutham Ravipati, Li-Fan Wong, Sally Cole, Tushar D. Bhagat, Jonathan Feld, Yosman Dhar, Matthias Bartenstein, Victor J. Thiruthuvanathan, Amittha Wickrema, B. Hilda Ye, David A. Frank, Andrea Pellagatti, Jacqueline Boultwood, Tianyuan Zhou, Youngsoo Kim, A. Robert MacLeod, P.K. Epling-Burnette, Minwei Ye, Patricia McCoon, Richard Woessner, Ulrich Steidl, Britta Will, Amit Verma
Aditi Shastri, Gaurav Choudhary, Margarida Teixeira, Shanisha Gordon-Mitchell, Nandini Ramachandra, Lumie Bernard, Sanchari Bhattacharyya, Robert Lopez, Kith Pradhan, Orsolya Giricz, Goutham Ravipati, Li-Fan Wong, Sally Cole, Tushar D. Bhagat, Jonathan Feld, Yosman Dhar, Matthias Bartenstein, Victor J. Thiruthuvanathan, Amittha Wickrema, B. Hilda Ye, David A. Frank, Andrea Pellagatti, Jacqueline Boultwood, Tianyuan Zhou, Youngsoo Kim, A. Robert MacLeod, P.K. Epling-Burnette, Minwei Ye, Patricia McCoon, Richard Woessner, Ulrich Steidl, Britta Will, Amit Verma
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Research Article Hematology

Antisense STAT3 inhibitor decreases viability of myelodysplastic and leukemic stem cells

Abstract

Acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS) are associated with disease-initiating stem cells that are not eliminated by conventional therapies. Transcriptomic analysis of stem and progenitor populations in MDS and AML demonstrated overexpression of STAT3 that was validated in an independent cohort. STAT3 overexpression was predictive of a shorter survival and worse clinical features in a large MDS cohort. High STAT3 expression signature in MDS CD34+ cells was similar to known preleukemic gene signatures. Functionally, STAT3 inhibition by a clinical, antisense oligonucleotide, AZD9150, led to reduced viability and increased apoptosis in leukemic cell lines. AZD9150 was rapidly incorporated by primary MDS/AML stem and progenitor cells and led to increased hematopoietic differentiation. STAT3 knockdown also impaired leukemic growth in vivo and led to decreased expression of MCL1 and other oncogenic genes in malignant cells. These studies demonstrate that STAT3 is an adverse prognostic factor in MDS/AML and provide a preclinical rationale for studies using AZD9150 in these diseases.

Authors

Aditi Shastri, Gaurav Choudhary, Margarida Teixeira, Shanisha Gordon-Mitchell, Nandini Ramachandra, Lumie Bernard, Sanchari Bhattacharyya, Robert Lopez, Kith Pradhan, Orsolya Giricz, Goutham Ravipati, Li-Fan Wong, Sally Cole, Tushar D. Bhagat, Jonathan Feld, Yosman Dhar, Matthias Bartenstein, Victor J. Thiruthuvanathan, Amittha Wickrema, B. Hilda Ye, David A. Frank, Andrea Pellagatti, Jacqueline Boultwood, Tianyuan Zhou, Youngsoo Kim, A. Robert MacLeod, P.K. Epling-Burnette, Minwei Ye, Patricia McCoon, Richard Woessner, Ulrich Steidl, Britta Will, Amit Verma

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Figure 3

AZD9150 specifically inhibits STAT3 and decreases viability and induces apoptosis in leukemic cell lines.

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AZD9150 specifically inhibits STAT3 and decreases viability and induces ...
(A) To demonstrate specific inhibition of STAT3-mediated gene expression, we treated the CMK cell lines transfected with STAT3- and STAT5-binding luciferase constructs with AZD9150 (2.5 μM) and control. A decrease in STAT3-mediated luciferase expression was observed, while no change in STAT5-mediated gene expression was seen. (B) qPCR of 4 leukemic cell lines transfected with AZD9150 (10 μM) showed a decreased expression of STAT3. (C) Multiple leukemic cells lines were transfected with AZD9150 (10 μM) and inactive structural analog control (10 μM). AZD9150 resulted in a significant decrease in viable cells. (D) Representative flow cytometry plots show increased apoptosis with AZD9150 treatment (5 μM) compared with the control oligonucleotide (5 μM). (E–G) Increased apoptosis was seen in multiple leukemic cell lines after AZD9150 transfection as compared with controls. (H) NOD-SCID mice were injected with CMK cells transfected with AZD9150 (10 μM) and the control oligonucleotide (10 μM). The 2 cohorts were followed over time for survival, and it was noted that the drug-treated mice had an improved median survival of 64 days compared with 43 days for the control oligonucleotide (P = 0.028). *P ≤ 0.05, **P ≤ 0.005.