Autoinhibition of Bcr-Abl through Its SH3 Domain

Smith, Kristen M.; Yacobi, Rinat; Etten, Richard A. Van

doi:10.1016/s1097-2765(03)00274-0

Cited by 137 publications

(120 citation statements)

References 40 publications

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“…To compare the relative oncogenic potency of eight different kinase inhibitor resistant BCR-ABL p210 mutants (Fig. 1A), we used a modified Whitlock-Witte pre-B cell transformation assay (22)(23)(24)(25) that was previously shown to correlate with in vivo CML mouse models and can distinguish subtle and quantifiable differences in po-tency between different BCR-ABL isoforms (26). These include two detected pretreatment in patients (T315I and E255K) and one documented to disappear after discontinuation of kinase inhibitor therapy (Y253H) (27).…”

Section: Transformation Potency Of Most Kinase Inhibitor-resistant Bcmentioning

confidence: 99%

Phosphorylation of the ATP-binding loop directs oncogenicity of drug-resistant BCR-ABL mutants

Skaggs

Gorre²,

Рывкин³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

128

140

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The success of targeting kinases in cancer with small molecule inhibitors has been tempered by the emergence of drug-resistant kinase domain mutations. In patients with chronic myeloid leukemia treated with ABL inhibitors, BCR-ABL kinase domain mutations are the principal mechanism of relapse. Certain mutations are occasionally detected before treatment, suggesting increased fitness relative to wild-type p210 BCR-ABL. We evaluated the oncogenicity of eight kinase inhibitor-resistant BCR-ABL mutants and found a spectrum of potencies greater or less than p210. Although most fitness alterations correlate with changes in kinase activity, this is not the case with the T315I BCR-ABL mutation that confers clinical resistance to all currently approved ABL kinase inhibitors. Through global phosphoproteome analysis, we identified a unique phosphosubstrate signature associated with each drug-resistant allele, including a shift in phosphorylation of two tyrosines (Tyr 253 and Tyr 257 ) in the ATP binding loop (P-loop) of BCR-ABL when Thr 315 is Ile or Ala. Mutational analysis of these tyrosines in the context of Thr 315 mutations demonstrates that the identity of the gatekeeper residue impacts oncogenicity by altered P-loop phosphorylation. Therefore, mutations that confer clinical resistance to kinase inhibitors can substantially alter kinase function and confer novel biological properties that may impact disease progression.chronic myelogenous leukemia ͉ kinase inhibitor resistance ͉ phosphoproteomics ͉ imatinib ͉ dasatinib P oint mutations in the kinase domain of BCR-ABL are primarily responsible for resistance to the ABL inhibitor imatinib (Gleevec) in chronic myelogenous leukemia (CML) patients. The majority of imatinib-resistant BCR-ABL point mutations (of Ͼ50 distinct examples now reported clinically) impair drug binding by restricting flexibility of the enzyme, precluding adoption of the inactive conformation required for imatinib binding (1-4). The second generation Abl inhibitor dasatinib is effective against imatinib-resistant CML because it binds the BCR-ABL kinase domain regardless of activation loop conformation (5-7). As a result, the number of BCR-ABL mutations capable of conferring resistance to dasatinib is small and is limited almost exclusively to direct contact sites (8, 9). One mutation, T315I, confers resistance to imatinib, dasatinib, and the imatinib-related compound nilotinib (AMN-107) (10, 11).Although discovered in the context of drug resistance, there is growing evidence that these mutations may confer other fitness advantages to BCR-ABL. First, the T315I and E255K mutants were each detected by our group in imatinib-naïve CML blast crisis patients by direct sequencing of total BCR-ABL cDNA and are therefore estimated to account for at least 20% of the CML tumor burden in these patients (1). In addition, these and other kinase domain mutations have been identified before treatment in CML patients using mutation-specific quantitative PCR (12-18). Furthermore, the analogous mutation to T315I in the...

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Section: Transformation Potency Of Most Kinase Inhibitor-resistant Bcmentioning

confidence: 99%

Phosphorylation of the ATP-binding loop directs oncogenicity of drug-resistant BCR-ABL mutants

Skaggs

Gorre²,

Рывкин³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

128

140

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“…The amino acid sequence of Tel bears no homology to Bcr, but the Tel PNT domain mediates oligomerization (13,26) and is required for dysregulated in vivo kinase activity (13) and induction of myeloproliferative disease in mice (38) by Tel-Abl. This suggests that oligomerization of Abl is a critical step in the dysregulation of Abl kinase activity leading to leukemia (54,55). However, oligomerization alone appears to be insufficient for full oncogenic activation of Abl, because fusion of just the Bcr oligomerization domain to c-Abl yields proteins that are defective for transformation (37) and leukemogenesis (21,63).…”

Section: Discussionmentioning

confidence: 99%

“…The different retroviral stocks demonstrated equivalent transduction of primary bone marrow by Southern blot determination of the proviral DNA content in genomic DNA (data not shown). Immediately after transduction, cells were plated for in vitro growth in Whitlock/Witte-style cultures as described previously (55) or transplanted into irradiated (2 ϫ 450 cGy) syngeneic female recipient mice (10 6 cells each). Whitlock/Witte cultures were performed in triplicate in 24-well plates at 1 ϫ 10 6 , 3 ϫ 10 5 , 1 ϫ 10 5 , 3 ϫ 10 4 , 1 ϫ 10 4 , 3 ϫ 10 3 , 1 ϫ 10 5 , or 3 ϫ 10 2 transduced cells per well in RPMI 1640 medium supplemented with 10% fetal calf serum, 200 M L-glutamine, 50 M 2-mercaptoethanol, and penicillinstreptomycin.…”

Section: Signal Transduction Analysismentioning

confidence: 99%

A Direct Binding Site for Grb2 Contributes to Transformation and Leukemogenesis by the Tel-Abl (ETV6-Abl) Tyrosine Kinase

Million

Harakawa

Rumyantsev

et al. 2004

Molecular and Cellular Biology

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A direct binding site for the Grb2 adapter protein is required for the induction of fatal chronic myeloid leukemia (CML)-like disease in mice by Bcr-Abl. Here, we demonstrate direct binding of Grb2 to the Tel-Abl (ETV6-Abl) fusion protein, the product of complex (9;12) chromosomal translocations in human leukemia, via tyrosine 314 encoded by TEL exon 5. A Tel-Abl point mutant (Y314F) and a splice variant without TEL exon 5 sequences (⌬e5) lacked Grb2 interaction and exhibited decreased binding and phosphorylation of the scaffolding protein Gab2 and impaired activation of phosphatidylinositol 3-kinase, Akt, and extracellular signal-regulated kinase/mitogen-activated protein kinase in hematopoietic cells. Tel-Abl Y314F and ⌬e5 were unable to transform fibroblasts to anchorage-independent growth and were defective for B-lymphoid transformation in vitro and lymphoid leukemogenesis in vivo. Previously, we demonstrated that full-length Tel-Abl induced two distinct myeloproliferative diseases in mice: CML-like leukemia similar to that induced by Bcr-Abl and a novel syndrome of small-bowel myeloid infiltration endotoxemia and hepatic and renal failure. Lack of the Grb2 binding site had no effect on development of small bowel syndrome but significantly attenuated the induction of CML-like disease by Tel-Abl. These results suggest that direct binding of Grb2 is a common mechanism contributing to leukemogenesis by oncogenic Abl fusion proteins.The BCR-ABL oncogene, the product of the t(9;22) Philadelphia (Ph) chromosome translocation, encodes a dysregulated cytoplasmic protein-tyrosine kinase, Bcr-Abl, that is the direct cause of the myeloproliferative disease chronic myeloid leukemia (CML) and Ph ϩ acute B-lymphoblastic leukemia (B-ALL). Bcr-Abl activates multiple intracellular signaling pathways including Ras, mitogen-activated protein kinase (MAPK), Jun N-terminal kinase (JNK), STAT5, and phosphatidylinositol 3-kinase (PI 3-kinase) (52) and transforms fibroblasts (33), cytokine-dependent hematopoietic cell lines (6,18), and primary bone marrow B-lymphoid cells (36) in vitro. Retroviral transduction of the BCR-ABL gene into murine bone marrow followed by transplantation into irradiated recipient mice results in the development of either CML-like myeloproliferative disease (30,44,62) or B-ALL (50) in all recipients, depending on the transduction conditions. The mouse retroviral bone marrow transduction/transplantation system provides accurate and quantitative models of human CML and Ph ϩ B-ALL (58) that have proven useful for analyzing the molecular pathophysiology of these diseases (15,29,30,39,50,63).Fusion of the ABL gene to a different partner, the TEL (ETV6) gene on chromosome 12p13, has been reported to occur in a small number of patients with leukemia, some who had acute leukemia of B-lymphoid (43), T-lymphoid (60), or myeloid (13, 40) origin and some who presented with atypical (3, 28) or typical (1, 60) CML. TEL encodes a ubiquitously expressed 452-amino-acid protein with homology to the Ets family of transcripti...

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“…A mutant form of BCR-ABL that lacks the BCR-CC domain (ΔCC-BCR-ABL) failed to induce MPD in mice, but, rather, induced a T-cell leukemia/ lymphoma only after a long latent period (Zhang et al, 2001;Smith et al, 2003). Reactivation of the kinase activity of ABL by mutating its SH3 domain (through deletion or a P1013L point mutation), rescued the ability of ΔCC-BCR-ABL to induce a CML-like MPD in mice (Smith et al, 2003). These results demonstrate that the BCR domain is essential for the induction of CML by BCR-ABL in mice, mainly owing to its ability to activate the kinase activity of ABL.…”

Section: Bcr-abl Domain Functions and CML Mouse Modelmentioning

confidence: 99%

“…Mutations in the SH2 domain of ABL reduced the ability of BCR-ABL to induce a CML-like MPD in mice (Zhang et al, 2001). The Y1294F point mutation in SH2 domain of BCR-ABL also attenuated leukemogenesis by BCR-ABL (Smith et al, 2003). The carboxy-terminal region of ABL is required for the proper function of normal ABL and for the lymphoid leukemogenicity of v-Abl (Prywes et al, 1983).…”

Section: Bcr-abl Domain Functions and CML Mouse Modelmentioning

confidence: 99%

Molecular and cellular bases of chronic myeloid leukemia

Chen¹,

Peng²,

Li³

et al. 2010

Protein Cell

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Chronic myeloid leukemia (CML) is a myeloproliferative disease characterized by the overproduction of granulocytes, which leads to high white blood cell counts and splenomegaly in patients. Based on clinical symptoms and laboratory findings, CML is classified into three clinical phases, often starting with a chronic phase, progressing to an accelerated phase and ultimately ending in a terminal phase called blast crisis. Blast crisis phase of CML is clinically similar to an acute leukemia; in particular, B-cell acute lymphoblastic leukemia (B-ALL) is a severe form of acute leukemia in blast crisis, and there is no effective therapy for it yet. CML is induced by the BCR-ABL oncogene, whose gene product is a BCR-ABL tyrosine kinase. Currently, inhibition of BCR-ABL kinase activity by its kinase inhibitor such as imatinib mesylate (Gleevec) is a major therapeutic strategy for CML. However, the inability of BCR-ABL kinase inhibitors to completely kill leukemia stem cells (LSCs) indicates that these kinase inhibitors are unlikely to cure CML. In addition, drug resistance due to the development of BCR-ABL mutations occurs before and during treatment of CML with kinase inhibitors. A critical issue to resolve this problem is to fully understand the biology of LSCs, and to identify key genes that play significant roles in survival and self-renewal of LSCs. In this review, we will focus on LSCs in CML by summarizing and discussing available experimental results, including the original studies from our own laboratory.

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Autoinhibition of Bcr-Abl through Its SH3 Domain

Cited by 137 publications

References 40 publications

Phosphorylation of the ATP-binding loop directs oncogenicity of drug-resistant BCR-ABL mutants

Phosphorylation of the ATP-binding loop directs oncogenicity of drug-resistant BCR-ABL mutants

A Direct Binding Site for Grb2 Contributes to Transformation and Leukemogenesis by the Tel-Abl (ETV6-Abl) Tyrosine Kinase

Molecular and cellular bases of chronic myeloid leukemia

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