Targeting BCR-ABL1 in chronic myeloid leukemia by PROTAC-mediated targeted protein degradation

George M. Burslem, Anna Reister Schultz, Daniel P. Bondeson, Christopher A. Eide, Samantha L.Savage Stevens, Brian Druker, Craig M. Crews

Research output: Contribution to journalArticle

Abstract

Although the use of ATP-competitive tyrosine kinase inhibitors of oncoprotein BCR-ABL1 has enabled durable responses in patients with chronic myeloid leukemia (CML), issues of drug resistance and residual leukemic stem cells remain. To test whether the degradation of BCRABL1 kinase could offer improved response, we developed a series of proteolysis-targeting chimera (PROTAC) that allosterically target BCR-ABL1 protein and recruit the E3 ligase Von Hippel-Lindau, resulting in ubiquitination and subsequent degradation of the oncogenic fusion protein. In both human CML K562 cells and murine Ba/F3 cells expressing BCR-ABL1, lead compound GMB-475 induced rapid proteasomal degradation and inhibition of downstream biomarkers, such as STAT5, and showed increased sensitivity compared with diastereomeric controls lacking degradation activity. Notably, GMB-475 inhibited the proliferation of certain clinically relevant BCR-ABL1 kinase domain point mutants and further sensitized Ba/ F3 BCR-ABL1 cells to inhibition by imatinib, while demonstrating no toxicity toward Ba/F3 parental cells. Reverse phase protein array analysis suggested additional differences in levels of phosphorylated SHP2, GAB2, and SHC associated with BCR-ABL1 degradation. Importantly, GMB-475 reduced viability and increased apoptosis in primary CML CD34+ cells, with no effect on healthy CD34+ cells at identical concentrations. GMB-475 degraded BCR-ABL1 and reduced cell viability in primary CML stem cells. Together, these findings suggest that combined BCR-ABL1 kinase inhibition and protein degradation may represent a strategy to address BCR-ABL1-dependent drug resistance, and warrant further investigation into the eradication of persistent leukemic stem cells, which rely on neither the presence nor the activity of the BCR-ABL1 protein for survival. Significance: Small-molecule-induced degradation of BCR-ABL1 in CML provides an advantage over inhibition and provides insights into CML stem cell biology.

Original languageEnglish (US)
Pages (from-to)4744-4753
Number of pages10
JournalCancer Research
Volume79
Issue number18
DOIs
StatePublished - Sep 15 2019

Fingerprint

Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Proteolysis
Myeloid Progenitor Cells
Phosphotransferases
Drug Resistance
Stem Cells
Protein Array Analysis
Proteins
Ubiquitin-Protein Ligases
K562 Cells
Ubiquitination
Oncogene Proteins
Protein-Tyrosine Kinases
Cell Biology
Cell Survival
Adenosine Triphosphate
Biomarkers
Apoptosis
Survival

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Burslem, G. M., Schultz, A. R., Bondeson, D. P., Eide, C. A., Stevens, S. L. S., Druker, B., & Crews, C. M. (2019). Targeting BCR-ABL1 in chronic myeloid leukemia by PROTAC-mediated targeted protein degradation. Cancer Research, 79(18), 4744-4753. https://doi.org/10.1158/0008-5472.CAN-19-1236

Targeting BCR-ABL1 in chronic myeloid leukemia by PROTAC-mediated targeted protein degradation. / Burslem, George M.; Schultz, Anna Reister; Bondeson, Daniel P.; Eide, Christopher A.; Stevens, Samantha L.Savage; Druker, Brian; Crews, Craig M.

In: Cancer Research, Vol. 79, No. 18, 15.09.2019, p. 4744-4753.

Research output: Contribution to journalArticle

Burslem, GM, Schultz, AR, Bondeson, DP, Eide, CA, Stevens, SLS, Druker, B & Crews, CM 2019, 'Targeting BCR-ABL1 in chronic myeloid leukemia by PROTAC-mediated targeted protein degradation', Cancer Research, vol. 79, no. 18, pp. 4744-4753. https://doi.org/10.1158/0008-5472.CAN-19-1236
Burslem, George M. ; Schultz, Anna Reister ; Bondeson, Daniel P. ; Eide, Christopher A. ; Stevens, Samantha L.Savage ; Druker, Brian ; Crews, Craig M. / Targeting BCR-ABL1 in chronic myeloid leukemia by PROTAC-mediated targeted protein degradation. In: Cancer Research. 2019 ; Vol. 79, No. 18. pp. 4744-4753.
@article{9263aafffbfd437f972a2dbeafae88d7,
title = "Targeting BCR-ABL1 in chronic myeloid leukemia by PROTAC-mediated targeted protein degradation",
abstract = "Although the use of ATP-competitive tyrosine kinase inhibitors of oncoprotein BCR-ABL1 has enabled durable responses in patients with chronic myeloid leukemia (CML), issues of drug resistance and residual leukemic stem cells remain. To test whether the degradation of BCRABL1 kinase could offer improved response, we developed a series of proteolysis-targeting chimera (PROTAC) that allosterically target BCR-ABL1 protein and recruit the E3 ligase Von Hippel-Lindau, resulting in ubiquitination and subsequent degradation of the oncogenic fusion protein. In both human CML K562 cells and murine Ba/F3 cells expressing BCR-ABL1, lead compound GMB-475 induced rapid proteasomal degradation and inhibition of downstream biomarkers, such as STAT5, and showed increased sensitivity compared with diastereomeric controls lacking degradation activity. Notably, GMB-475 inhibited the proliferation of certain clinically relevant BCR-ABL1 kinase domain point mutants and further sensitized Ba/ F3 BCR-ABL1 cells to inhibition by imatinib, while demonstrating no toxicity toward Ba/F3 parental cells. Reverse phase protein array analysis suggested additional differences in levels of phosphorylated SHP2, GAB2, and SHC associated with BCR-ABL1 degradation. Importantly, GMB-475 reduced viability and increased apoptosis in primary CML CD34+ cells, with no effect on healthy CD34+ cells at identical concentrations. GMB-475 degraded BCR-ABL1 and reduced cell viability in primary CML stem cells. Together, these findings suggest that combined BCR-ABL1 kinase inhibition and protein degradation may represent a strategy to address BCR-ABL1-dependent drug resistance, and warrant further investigation into the eradication of persistent leukemic stem cells, which rely on neither the presence nor the activity of the BCR-ABL1 protein for survival. Significance: Small-molecule-induced degradation of BCR-ABL1 in CML provides an advantage over inhibition and provides insights into CML stem cell biology.",
author = "Burslem, {George M.} and Schultz, {Anna Reister} and Bondeson, {Daniel P.} and Eide, {Christopher A.} and Stevens, {Samantha L.Savage} and Brian Druker and Crews, {Craig M.}",
year = "2019",
month = "9",
day = "15",
doi = "10.1158/0008-5472.CAN-19-1236",
language = "English (US)",
volume = "79",
pages = "4744--4753",
journal = "Journal of Cancer Research",
issn = "0099-7013",
publisher = "American Association for Cancer Research Inc.",
number = "18",

}

TY - JOUR

T1 - Targeting BCR-ABL1 in chronic myeloid leukemia by PROTAC-mediated targeted protein degradation

AU - Burslem, George M.

AU - Schultz, Anna Reister

AU - Bondeson, Daniel P.

AU - Eide, Christopher A.

AU - Stevens, Samantha L.Savage

AU - Druker, Brian

AU - Crews, Craig M.

PY - 2019/9/15

Y1 - 2019/9/15

N2 - Although the use of ATP-competitive tyrosine kinase inhibitors of oncoprotein BCR-ABL1 has enabled durable responses in patients with chronic myeloid leukemia (CML), issues of drug resistance and residual leukemic stem cells remain. To test whether the degradation of BCRABL1 kinase could offer improved response, we developed a series of proteolysis-targeting chimera (PROTAC) that allosterically target BCR-ABL1 protein and recruit the E3 ligase Von Hippel-Lindau, resulting in ubiquitination and subsequent degradation of the oncogenic fusion protein. In both human CML K562 cells and murine Ba/F3 cells expressing BCR-ABL1, lead compound GMB-475 induced rapid proteasomal degradation and inhibition of downstream biomarkers, such as STAT5, and showed increased sensitivity compared with diastereomeric controls lacking degradation activity. Notably, GMB-475 inhibited the proliferation of certain clinically relevant BCR-ABL1 kinase domain point mutants and further sensitized Ba/ F3 BCR-ABL1 cells to inhibition by imatinib, while demonstrating no toxicity toward Ba/F3 parental cells. Reverse phase protein array analysis suggested additional differences in levels of phosphorylated SHP2, GAB2, and SHC associated with BCR-ABL1 degradation. Importantly, GMB-475 reduced viability and increased apoptosis in primary CML CD34+ cells, with no effect on healthy CD34+ cells at identical concentrations. GMB-475 degraded BCR-ABL1 and reduced cell viability in primary CML stem cells. Together, these findings suggest that combined BCR-ABL1 kinase inhibition and protein degradation may represent a strategy to address BCR-ABL1-dependent drug resistance, and warrant further investigation into the eradication of persistent leukemic stem cells, which rely on neither the presence nor the activity of the BCR-ABL1 protein for survival. Significance: Small-molecule-induced degradation of BCR-ABL1 in CML provides an advantage over inhibition and provides insights into CML stem cell biology.

AB - Although the use of ATP-competitive tyrosine kinase inhibitors of oncoprotein BCR-ABL1 has enabled durable responses in patients with chronic myeloid leukemia (CML), issues of drug resistance and residual leukemic stem cells remain. To test whether the degradation of BCRABL1 kinase could offer improved response, we developed a series of proteolysis-targeting chimera (PROTAC) that allosterically target BCR-ABL1 protein and recruit the E3 ligase Von Hippel-Lindau, resulting in ubiquitination and subsequent degradation of the oncogenic fusion protein. In both human CML K562 cells and murine Ba/F3 cells expressing BCR-ABL1, lead compound GMB-475 induced rapid proteasomal degradation and inhibition of downstream biomarkers, such as STAT5, and showed increased sensitivity compared with diastereomeric controls lacking degradation activity. Notably, GMB-475 inhibited the proliferation of certain clinically relevant BCR-ABL1 kinase domain point mutants and further sensitized Ba/ F3 BCR-ABL1 cells to inhibition by imatinib, while demonstrating no toxicity toward Ba/F3 parental cells. Reverse phase protein array analysis suggested additional differences in levels of phosphorylated SHP2, GAB2, and SHC associated with BCR-ABL1 degradation. Importantly, GMB-475 reduced viability and increased apoptosis in primary CML CD34+ cells, with no effect on healthy CD34+ cells at identical concentrations. GMB-475 degraded BCR-ABL1 and reduced cell viability in primary CML stem cells. Together, these findings suggest that combined BCR-ABL1 kinase inhibition and protein degradation may represent a strategy to address BCR-ABL1-dependent drug resistance, and warrant further investigation into the eradication of persistent leukemic stem cells, which rely on neither the presence nor the activity of the BCR-ABL1 protein for survival. Significance: Small-molecule-induced degradation of BCR-ABL1 in CML provides an advantage over inhibition and provides insights into CML stem cell biology.

UR - http://www.scopus.com/inward/record.url?scp=85072233649&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85072233649&partnerID=8YFLogxK

U2 - 10.1158/0008-5472.CAN-19-1236

DO - 10.1158/0008-5472.CAN-19-1236

M3 - Article

VL - 79

SP - 4744

EP - 4753

JO - Journal of Cancer Research

JF - Journal of Cancer Research

SN - 0099-7013

IS - 18

ER -