A Pan-BCL2 inhibitor renders bone-marrow-resident human leukemia stem cells sensitive to tyrosine kinase inhibition

Daniel J. Goff; Angela Court Recart; Anil Sadarangani; Hye Jung Chun; Christian L. Barrett; Maryla Krajewska; Heather Leu; Janine Low-Marchelli; Wenxue Ma; Alice Y. Shih; Jun Wei; Dayong Zhai; Ifat Geron; Minya Pu; Lei Bao; Ryan Chuang; Larisa Balaian; Jason Gotlib; Mark Minden; Giovanni Martinelli; Jessica Rusert; Kim Hien Dao; Kamran Shazand; Peggy Wentworth; Kristen M. Smith; Christina A.M. Jamieson; Sheldon R. Morris; Karen Messer; Lawrence S.B. Goldstein; Thomas J. Hudson; Marco Marra; Kelly A. Frazer; Maurizio Pellecchia; John C. Reed; Catriona H.M. Jamieson

doi:10.1016/j.stem.2012.12.011

A Pan-BCL2 inhibitor renders bone-marrow-resident human leukemia stem cells sensitive to tyrosine kinase inhibition

Daniel J. Goff, Angela Court Recart, Anil Sadarangani, Hye Jung Chun, Christian L. Barrett, Maryla Krajewska, Heather Leu, Janine Low-Marchelli, Wenxue Ma, Alice Y. Shih, Jun Wei, Dayong Zhai, Ifat Geron, Minya Pu, Lei Bao, Ryan Chuang, Larisa Balaian, Jason Gotlib, Mark Minden, Giovanni MartinelliJessica Rusert, Kim Hien Dao, Kamran Shazand, Peggy Wentworth, Kristen M. Smith, Christina A.M. Jamieson, Sheldon R. Morris, Karen Messer, Lawrence S.B. Goldstein, Thomas J. Hudson, Marco Marra, Kelly A. Frazer, Maurizio Pellecchia, John C. Reed, Catriona H.M. Jamieson^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

162 Scopus citations

Abstract

Leukemia stem cells (LSCs) play a pivotal role in the resistance of chronic myeloid leukemia (CML) to tyrosine kinase inhibitors (TKIs) and its progression to blast crisis (BC), in part, through the alternative splicing of self-renewal and survival genes. To elucidate splice-isoform regulators of human BC LSC maintenance, we performed whole-transcriptome RNA sequencing, splice-isoform-specific quantitative RT-PCR (qRT-PCR), nanoproteomics, stromal coculture, and BC LSC xenotransplantation analyses. Cumulatively, these studies show that the alternative splicing of multiple prosurvival BCL2 family genes promotes malignant transformation of myeloid progenitors into BC LSCS that are quiescent in the marrow niche and that contribute to therapeutic resistance. Notably, sabutoclax, a pan-BCL2 inhibitor, renders marrow-niche-resident BC LSCs sensitive to TKIs at doses that spare normal progenitors. These findings underscore the importance of alternative BCL2 family splice-isoform expression in BC LSC maintenance and suggest that the combinatorial inhibition of prosurvival BCL2 family proteins and BCR-ABL may eliminate dormant LSCs and obviate resistance.

Original language	English
Pages (from-to)	316-328
Number of pages	13
Journal	Cell Stem Cell
Volume	12
Issue number	3
DOIs	https://doi.org/10.1016/j.stem.2012.12.011
State	Published - 7 Mar 2013
Externally published	Yes

Bibliographical note

Funding Information:
We thank Dennis Carson for his continuous advice and mentorship, Dennis Young for his expert assistance on all FACS experiments, Ida Deichaite for her excellent assistance with material transfer and industry relations, Jennifer Black and Rusty Wall for their help with sample preparation and mouse experiments, Isabel Newton for her advice and help with DiR studies, Derrick Duarte for his assistance with immunohistochemistry studies, Jerry Wu for his assistance with FACS cell-cycle experiments, and Kimberly Wilson for her assistance with grant and manuscript preparation and submission. This work was generously supported by the Ratner Family Fund and by the California Institute for Regenerative Medicine (CIRM; grants RN2-00910-1, RS1-00228-1, TR2-01789, and DR1-01430). D.J.G. was supported by the CIRM UCSD Stem Cell Training Grant II and the UCSD Cancer Training Grant. This work was also funded by the National Cancer Institute (NCI; no. CA-55164) and the National Institutes of Health (NIH; no. CA-149668), and supported by the Ontario Institute for Cancer Research, through generous support from the Ontario Ministry of Research and Innovation and the Cancer Stem Cell Consortium with funding from the Government of Canada through Genome Canada and the Ontario Genomics Institute (OGI-047), and through the Canadian Institute of Health Research (CSC-105367). J.C.R., J.W., and M.P. are coinventors of sabutoclax and related compounds, licensed by the SBMRI to Oncothyreon (Seattle). The Oncothyreon license entitles J.C.R. to milestone and royalty payments for studies advancing to clinic. He does not receive consulting fees or lab support, nor does he have stock or stock options.

Keywords

Blast Crisis
Gossypol
Humans
Leukemia
Neoplastic Stem Cells
Protein Kinase Inhibitors
Proto-Oncogene Proteins c-bcl-2
Reverse Transcriptase Polymerase Chain Reaction

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10.1016/j.stem.2012.12.011

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Goff, D. J., Recart, A. C., Sadarangani, A., Chun, H. J., Barrett, C. L., Krajewska, M., Leu, H., Low-Marchelli, J., Ma, W., Shih, A. Y., Wei, J., Zhai, D., Geron, I., Pu, M., Bao, L., Chuang, R., Balaian, L., Gotlib, J., Minden, M., ... Jamieson, C. H. M. (2013). A Pan-BCL2 inhibitor renders bone-marrow-resident human leukemia stem cells sensitive to tyrosine kinase inhibition. Cell Stem Cell, 12(3), 316-328. https://doi.org/10.1016/j.stem.2012.12.011

@article{f7fe1a19ddb245938ffc62bb241e4d19,

title = "A Pan-BCL2 inhibitor renders bone-marrow-resident human leukemia stem cells sensitive to tyrosine kinase inhibition",

abstract = "Leukemia stem cells (LSCs) play a pivotal role in the resistance of chronic myeloid leukemia (CML) to tyrosine kinase inhibitors (TKIs) and its progression to blast crisis (BC), in part, through the alternative splicing of self-renewal and survival genes. To elucidate splice-isoform regulators of human BC LSC maintenance, we performed whole-transcriptome RNA sequencing, splice-isoform-specific quantitative RT-PCR (qRT-PCR), nanoproteomics, stromal coculture, and BC LSC xenotransplantation analyses. Cumulatively, these studies show that the alternative splicing of multiple prosurvival BCL2 family genes promotes malignant transformation of myeloid progenitors into BC LSCS that are quiescent in the marrow niche and that contribute to therapeutic resistance. Notably, sabutoclax, a pan-BCL2 inhibitor, renders marrow-niche-resident BC LSCs sensitive to TKIs at doses that spare normal progenitors. These findings underscore the importance of alternative BCL2 family splice-isoform expression in BC LSC maintenance and suggest that the combinatorial inhibition of prosurvival BCL2 family proteins and BCR-ABL may eliminate dormant LSCs and obviate resistance.",

keywords = "Blast Crisis, Gossypol, Humans, Leukemia, Neoplastic Stem Cells, Protein Kinase Inhibitors, Proto-Oncogene Proteins c-bcl-2, Reverse Transcriptase Polymerase Chain Reaction",

author = "Goff, {Daniel J.} and Recart, {Angela Court} and Anil Sadarangani and Chun, {Hye Jung} and Barrett, {Christian L.} and Maryla Krajewska and Heather Leu and Janine Low-Marchelli and Wenxue Ma and Shih, {Alice Y.} and Jun Wei and Dayong Zhai and Ifat Geron and Minya Pu and Lei Bao and Ryan Chuang and Larisa Balaian and Jason Gotlib and Mark Minden and Giovanni Martinelli and Jessica Rusert and Dao, {Kim Hien} and Kamran Shazand and Peggy Wentworth and Smith, {Kristen M.} and Jamieson, {Christina A.M.} and Morris, {Sheldon R.} and Karen Messer and Goldstein, {Lawrence S.B.} and Hudson, {Thomas J.} and Marco Marra and Frazer, {Kelly A.} and Maurizio Pellecchia and Reed, {John C.} and Jamieson, {Catriona H.M.}",

note = "Funding Information: We thank Dennis Carson for his continuous advice and mentorship, Dennis Young for his expert assistance on all FACS experiments, Ida Deichaite for her excellent assistance with material transfer and industry relations, Jennifer Black and Rusty Wall for their help with sample preparation and mouse experiments, Isabel Newton for her advice and help with DiR studies, Derrick Duarte for his assistance with immunohistochemistry studies, Jerry Wu for his assistance with FACS cell-cycle experiments, and Kimberly Wilson for her assistance with grant and manuscript preparation and submission. This work was generously supported by the Ratner Family Fund and by the California Institute for Regenerative Medicine (CIRM; grants RN2-00910-1, RS1-00228-1, TR2-01789, and DR1-01430). D.J.G. was supported by the CIRM UCSD Stem Cell Training Grant II and the UCSD Cancer Training Grant. This work was also funded by the National Cancer Institute (NCI; no. CA-55164) and the National Institutes of Health (NIH; no. CA-149668), and supported by the Ontario Institute for Cancer Research, through generous support from the Ontario Ministry of Research and Innovation and the Cancer Stem Cell Consortium with funding from the Government of Canada through Genome Canada and the Ontario Genomics Institute (OGI-047), and through the Canadian Institute of Health Research (CSC-105367). J.C.R., J.W., and M.P. are coinventors of sabutoclax and related compounds, licensed by the SBMRI to Oncothyreon (Seattle). The Oncothyreon license entitles J.C.R. to milestone and royalty payments for studies advancing to clinic. He does not receive consulting fees or lab support, nor does he have stock or stock options. ",

year = "2013",

month = mar,

day = "7",

doi = "10.1016/j.stem.2012.12.011",

language = "English",

volume = "12",

pages = "316--328",

journal = "Cell Stem Cell",

issn = "1934-5909",

publisher = "Cell Press",

number = "3",

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Goff, DJ, Recart, AC, Sadarangani, A, Chun, HJ, Barrett, CL, Krajewska, M, Leu, H, Low-Marchelli, J, Ma, W, Shih, AY, Wei, J, Zhai, D, Geron, I, Pu, M, Bao, L, Chuang, R, Balaian, L, Gotlib, J, Minden, M, Martinelli, G, Rusert, J, Dao, KH, Shazand, K, Wentworth, P, Smith, KM, Jamieson, CAM, Morris, SR, Messer, K, Goldstein, LSB, Hudson, TJ, Marra, M, Frazer, KA, Pellecchia, M, Reed, JC & Jamieson, CHM 2013, 'A Pan-BCL2 inhibitor renders bone-marrow-resident human leukemia stem cells sensitive to tyrosine kinase inhibition', Cell Stem Cell, vol. 12, no. 3, pp. 316-328. https://doi.org/10.1016/j.stem.2012.12.011

TY - JOUR

T1 - A Pan-BCL2 inhibitor renders bone-marrow-resident human leukemia stem cells sensitive to tyrosine kinase inhibition

AU - Goff, Daniel J.

AU - Recart, Angela Court

AU - Sadarangani, Anil

AU - Chun, Hye Jung

AU - Barrett, Christian L.

AU - Krajewska, Maryla

AU - Leu, Heather

AU - Low-Marchelli, Janine

AU - Ma, Wenxue

AU - Shih, Alice Y.

AU - Wei, Jun

AU - Zhai, Dayong

AU - Geron, Ifat

AU - Pu, Minya

AU - Bao, Lei

AU - Chuang, Ryan

AU - Balaian, Larisa

AU - Gotlib, Jason

AU - Minden, Mark

AU - Martinelli, Giovanni

AU - Rusert, Jessica

AU - Dao, Kim Hien

AU - Shazand, Kamran

AU - Wentworth, Peggy

AU - Smith, Kristen M.

AU - Jamieson, Christina A.M.

AU - Morris, Sheldon R.

AU - Messer, Karen

AU - Goldstein, Lawrence S.B.

AU - Hudson, Thomas J.

AU - Marra, Marco

AU - Frazer, Kelly A.

AU - Pellecchia, Maurizio

AU - Reed, John C.

AU - Jamieson, Catriona H.M.

N1 - Funding Information: We thank Dennis Carson for his continuous advice and mentorship, Dennis Young for his expert assistance on all FACS experiments, Ida Deichaite for her excellent assistance with material transfer and industry relations, Jennifer Black and Rusty Wall for their help with sample preparation and mouse experiments, Isabel Newton for her advice and help with DiR studies, Derrick Duarte for his assistance with immunohistochemistry studies, Jerry Wu for his assistance with FACS cell-cycle experiments, and Kimberly Wilson for her assistance with grant and manuscript preparation and submission. This work was generously supported by the Ratner Family Fund and by the California Institute for Regenerative Medicine (CIRM; grants RN2-00910-1, RS1-00228-1, TR2-01789, and DR1-01430). D.J.G. was supported by the CIRM UCSD Stem Cell Training Grant II and the UCSD Cancer Training Grant. This work was also funded by the National Cancer Institute (NCI; no. CA-55164) and the National Institutes of Health (NIH; no. CA-149668), and supported by the Ontario Institute for Cancer Research, through generous support from the Ontario Ministry of Research and Innovation and the Cancer Stem Cell Consortium with funding from the Government of Canada through Genome Canada and the Ontario Genomics Institute (OGI-047), and through the Canadian Institute of Health Research (CSC-105367). J.C.R., J.W., and M.P. are coinventors of sabutoclax and related compounds, licensed by the SBMRI to Oncothyreon (Seattle). The Oncothyreon license entitles J.C.R. to milestone and royalty payments for studies advancing to clinic. He does not receive consulting fees or lab support, nor does he have stock or stock options.

PY - 2013/3/7

Y1 - 2013/3/7

N2 - Leukemia stem cells (LSCs) play a pivotal role in the resistance of chronic myeloid leukemia (CML) to tyrosine kinase inhibitors (TKIs) and its progression to blast crisis (BC), in part, through the alternative splicing of self-renewal and survival genes. To elucidate splice-isoform regulators of human BC LSC maintenance, we performed whole-transcriptome RNA sequencing, splice-isoform-specific quantitative RT-PCR (qRT-PCR), nanoproteomics, stromal coculture, and BC LSC xenotransplantation analyses. Cumulatively, these studies show that the alternative splicing of multiple prosurvival BCL2 family genes promotes malignant transformation of myeloid progenitors into BC LSCS that are quiescent in the marrow niche and that contribute to therapeutic resistance. Notably, sabutoclax, a pan-BCL2 inhibitor, renders marrow-niche-resident BC LSCs sensitive to TKIs at doses that spare normal progenitors. These findings underscore the importance of alternative BCL2 family splice-isoform expression in BC LSC maintenance and suggest that the combinatorial inhibition of prosurvival BCL2 family proteins and BCR-ABL may eliminate dormant LSCs and obviate resistance.

AB - Leukemia stem cells (LSCs) play a pivotal role in the resistance of chronic myeloid leukemia (CML) to tyrosine kinase inhibitors (TKIs) and its progression to blast crisis (BC), in part, through the alternative splicing of self-renewal and survival genes. To elucidate splice-isoform regulators of human BC LSC maintenance, we performed whole-transcriptome RNA sequencing, splice-isoform-specific quantitative RT-PCR (qRT-PCR), nanoproteomics, stromal coculture, and BC LSC xenotransplantation analyses. Cumulatively, these studies show that the alternative splicing of multiple prosurvival BCL2 family genes promotes malignant transformation of myeloid progenitors into BC LSCS that are quiescent in the marrow niche and that contribute to therapeutic resistance. Notably, sabutoclax, a pan-BCL2 inhibitor, renders marrow-niche-resident BC LSCs sensitive to TKIs at doses that spare normal progenitors. These findings underscore the importance of alternative BCL2 family splice-isoform expression in BC LSC maintenance and suggest that the combinatorial inhibition of prosurvival BCL2 family proteins and BCR-ABL may eliminate dormant LSCs and obviate resistance.

KW - Blast Crisis

KW - Gossypol

KW - Humans

KW - Leukemia

KW - Neoplastic Stem Cells

KW - Protein Kinase Inhibitors

KW - Proto-Oncogene Proteins c-bcl-2

KW - Reverse Transcriptase Polymerase Chain Reaction

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U2 - 10.1016/j.stem.2012.12.011

DO - 10.1016/j.stem.2012.12.011

M3 - Article

C2 - 23333150

AN - SCOPUS:84875131206

SN - 1934-5909

VL - 12

SP - 316

EP - 328

JO - Cell Stem Cell

JF - Cell Stem Cell

IS - 3

ER -

A Pan-BCL2 inhibitor renders bone-marrow-resident human leukemia stem cells sensitive to tyrosine kinase inhibition

Abstract

Bibliographical note

Keywords

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