|
|||||||||
|
|
|
|
|
|
|
|
|
|
|
Prepublished online as a Blood First Edition Paper on November 7, 2002; DOI 10.1182/blood-2002-05-1343.
NEOPLASIA
From the Department of Pathology and Kaplan
Comprehensive Cancer Center, and Department of Pediatric Oncology, New
York University School of Medicine, New York; Department of Pathology
and Centro di Ricerca in Medicina Sperimentale (CERMS), University of
Torino, Torino, Italy; Department of Pathology, University
of Verona, Verona, Italy; Department of Pathology,
University of Cornell, New York, NY.
Anaplastic Large Cell
Lymphomas (ALCLs) carry translocations in which the
anaplastic lymphoma kinase (ALK) gene is juxtaposed to various genes, the most common of which is the NPM/B23
gene. ALK fusion proteins result in the constitutive activation of ALK tyrosine kinase, thereby enhancing proliferation and increasing cell
survival. A direct role for NPM-ALK in cellular transformation has been
shown in vitro with immortalized cell lines and in vivo using
retroviral transfer experiments. Nonetheless, there is no direct
evidence of its oncogenic potential in T lymphocytes, which represent
the most common target of ALK chimeras. Here, we describe a new mouse
model of lymphomagenesis in which human NPM-ALK transcription was
targeted to T cells. NPM-ALK transgenic (Tg) mice were born with the
expected mendelian distribution, normal lymphoid organs, and a normal
number and proportion of helper and suppressor T cells. However, after
a short period of latency, all NPM-ALK Tg mice developed malignant
lymphoproliferative disorders (mean survival, 18 weeks). NPM-ALK Tg
thymic lymphomas displayed a T-cell phenotype characteristic of
immature thymocytes and frequently coexpressed surface CD30. A
subset of the NPM-ALK Tg mice also developed clonal B-cell plasma
cell neoplasms. These tumors arose in peripheral lymphoid organs
(plasmacytomas) or within the bone marrow and often led to peripheral
neuropathies and limb paralysis. Our NPM-ALK Tg mice are a suitable
model to dissect the molecular mechanisms of ALK-mediated
transformation and to investigate the efficacy of new therapeutic
approaches for the treatment of human ALCL in vivo.
(Blood. 2003;101:1919-1927) Human Anaplastic Large
Cell Lymphomas (ALCLs) are a unique subset of
lymphomas partly distinguished by their coexpression of the CD30
antigen.1 Classical cytogenetic studies demonstrated that
ALCLs carry unique translocations within the p23 region of chromosome
2.2-4 In 1994, Morris et al5 cloned the
t(2;5) translocation and discovered that a novel tyrosine kinase gene, the anaplastic lymphoma kinase (ALK), was fused to the
NPM/B23 gene. NPM participates in nucleocytoplasmic
trafficking6,7 and has been recently shown to regulate the
duplication of centrosomes.8 The ALK gene
encodes a tyrosine kinase receptor whose physiologic expression is
largely limited to neuronal cells.9,10 However, the
physiologic role of the ALK receptor remains largely unknown because
ALK In the past 5 years, several groups have successfully cloned new ALCL
translocations and demonstrated that the ALK gene can fuse
to multiple targets, which include the TFG, TPM3, ATIC, CLTCL, RanBP2, and MSN genes.11 Proteins fused to
ALK largely determine the subcellular localization of the derived
fusion proteins, being cytoplasmic (ATIC-, TGF-ALK, etc), cytoplasmic
and nuclear (NPM-ALK), or membranous (MSN-ALK).11
Moreover, ALK translocations can also be detected in nonlymphoid
neoplasms such as inflammatory myofibroblastic
tumors,14 and ALK expression has been described in
neuroblastomas15 as well as in a unique subtype of
immunoglobulin A (IgA)-positive plasmacytoid tumors.16
Cellular transformation by NPM-ALK has been demonstrated in
immortalized rodent fibroblasts17 and confirmed in studies
that have shown that ALK protects Ba/F3 and PC12 cells from
interleukin-3 or growth factor withdrawal13,17 (data not
shown). Transfer of NPM-ALK-transduced bone marrow cells into
irradiated host recipient mice resulted in the generation in vivo of
large cell B-cell lymphomas.18 In the past few years, the
molecular mechanisms of NPM-ALK-mediated cellular transformation have
also been partially elucidated.11 It has been shown that
the ALK portion of the fusion protein, corresponding to the cytoplasmic
tail of the ALK receptor and containing the catalytic domain, is
absolutely required for transformation,17 whereas all the
N-terminal regions of the ALK chimeras function as dimerization
domains.11,19 As a result of spontaneous dimerization, ALK
undergoes autophosphorylation and becomes catalytically active. Constitutively active ALK fusion proteins can bind multiple adaptor proteins and activate a series of pathways involved in cell
proliferation, transformation, and survival. These include the
phospholipase c To unveil the role of ALK in T-cell transformation, we generated a
mouse model in which expression of NPM-ALK was targeted to T
lymphocytes. All NPM-ALK transgenic (Tg) mice developed clonal lymphoproliferative disorders after a short period of latency. In
addition to T-cell lymphomas, a sizable fraction of our mice also
acquired plasma cell neoplasms. Studies using these NPM-ALK Tg mice
will allow a better understanding of the molecular mechanisms and
genetic defects leading to ALK-mediated transformation.
NPM-ALK Tg mice, cell lines, and statistical analysis
Positive NPM-ALK mice were detected by polymerase chain
reaction (PCR) by using genomic DNA obtained from mouse tail
biopsies as previously described.25 All experiments
presented in this study were derived from mice (C57BL/6 and Balb/c
backgrounds) obtained from 2 independent transgenic lines (N1 and N16).
Primary NPM-ALK cells were obtained from fresh thymic tumors
after being cultured in complete RPMI-1640 medium in vitro.
Survival curves were performed by using the nonparametric model of
Kaplan-Meier.
Immunoprecipitation and Western blot analysis
Southern blotting Southern blotting was performed as previously described.25 Briefly, high-molecular-weight genomic DNAs (10 µg) were digested by EcoRI, HindIII, or Pvu endonucleases, and then digested fragments were separated by electrophoresis. DNAs were subsequently transferred onto nitrocellulose. Radiolabeled cDNA probes were used to study the genomic configuration of T-cell receptor (TCR) and heavy-chain immunoglobulin loci.28 Human NPM-ALK genomic
sequences were investigated using BamHI-digested DNAs using
a specific ALK cDNA probed (BamHI-BamHI).
Flow cytometry, histology, and immunohistochemistry Single-cell suspensions were obtained from isolated tissue samples. Cells were washed, counted, and stained with the following murine primary fluorescein isothiocyanate (FITC)-, phycoerythrin (PE)-, or Tricolor-conjugated antibodies: Thy-1, CD4, CD8, B220, CD25, CD3, TCR /, TCR  / (Caltag Laboratories,
Burlingame, CA), CD30, CD44, and CD45RB (Pharmingen-BD Biosciences, San
Jose, CA). After staining (30 minutes at 4°C), cells were washed and analyzed by using a fluorescence-activated cell sorter scan (FACScan; Becton Dickinson) flow cytometer as described.25
For the histologic and immunohistochemistry analyses, tissue samples were fixed in PBS-buffered formalin (10%) and subsequently embedded in paraffin. Dewaxed 4-µm-thick tissue sections were stained with hematoxylin and eosin or after microwave retrieval (citrate buffer, pH 6.6, 15 minutes) incubated with anti-ALK primary antibody (1:1000; Zymed), anti-Ki-67 (Novacastra), anti-CD45R (B220; 1:100; Caltag), and anti-CD138 (1:20; Pharmingen-BD). Bound complexes were revealed by using the avidin biotin peroxidase complex and a semiautomated immunostainer (DAKO, Carpinteria, CA; or Ventana ES Medical Systems, Tucson, AZ). Mouse light- and heavy-chain expression was performed by using alkaline-conjugated rabbit antimouse antibodies (Southern Biotechnology Associates, Birmingham, AL). For immunofluorescence stains, paraffin-embedded tissue sections were treated as described earlier. Sections were then incubated with rabbit anti-ALK Ab. After washing, tissue sections were incubated with biotin-conjugated antirabbit Ab (1:200; Vector) and then FITC-Avidin (1:200; Sigma-Aldrich, St Louis, MO). Sections were subsequently incubated with normal rabbit serum (1:10, 30 minutes at RT) and then stained with PE-conjugated anti-B220 (1:20; Caltag) in presence of rabbit serum (1:10). After washing, slides were briefly dried and coverslipped with antifade (Vysis, Downers Grove, IL). Fluorescence staining was visualizes by using the 2.7 Cytovision software (Applied Imaging, Santa Clara, CA). Tissue culture The rate of spontaneous and in vitro-induced cell death was evaluated according to DNA content and propidium iodide or Annexin V (Pharmingen-BD) stainings.25 Briefly, thymocytes were cultivated with immobilized anti-CD3 antibody (10 µg/mL; 2C11; a gift of J. Bluestone) and soluble anti-CD28 antibody (5 µg/mL; Pharmingen-BD). Alternatively thymocytes were cocultured with dexamethasone (0.1 µM), tumor necrosis factor (TNF; 15 ng/mL), cycloheximide (75 µg/mL; Sigma-Aldrich), anti-Fas Ab (0.5 µg/mL; Pharmingen-BD), phorbol 12-myristate 13-acetate (PMA; 10 ng/mL; Sigma-Aldrich), or Ionomycin (1 µM; Sigma-Aldrich). At the indicated times, cells were harvested, washed, and stained.Purified peripheral T cells were obtained by magnetic-bead separation.
Briefly, 1 × 107 lymph node cells were first incubated
(for 30 minutes at RT) with a cocktail of antibodies (0.2-0.5 µg each
antibody/106 cells) against B cells (B220; Caltag),
macrophages (CD11c; Caltag), and natural killer (NK) cells (Anti-NK;
Caltag). At the end of incubation, 70 µL antirat-conjugated magnetic
beads (Dynabeads; Dynal, Lake Success, NY) were added. Bead-coated
cells were separated in a magnetic field, and unbound cells were washed
in cold PBS (3 times). Negatively selected T cells were first stained
with FITC-conjugated anti-CD3 mAb and analyzed by FACS to determine their purity (always > 95%). Highly purified T cells
(5 × 104) were cultivated in RPMI-1640 medium
supplemented with 10% fetal calf serum (FCS), streptomycin and
penicillin, and 10 Electrophoretic methods Semiatutomated agarose electrophoresis and immunofixation were performed on HYDRASYS and HYRYS systems (Sebia, Norcross, GA) according to the manufacturer's instructions. For protein electrophoresis, 10 µL sample was applied manually to the sample template. The subsequent sample application, electrophoresis (pH 8.6, 20 W, at 20°C), gel drying, and staining were performed automatically. The resulting electrophoretic profiles were scanned using the Hyris densitometer (Sebia). For immunofixation, each sample was applied in 6 different positions on agarose gels (Hydragel Immunofixation; Sebia), and the electrophoretic separation performed automatically under identical conditions as earlier. Either fixative or monospecific antisera to mouse immunoglobulins ( ,  , IgG, IgM, and IgA; Southern Biotechnology Associates) were applied to the electrophoresis lanes to
allow for fixation and immunoprecipitation, respectively. Detection of
monoclonal bands was assessed by visual inspection of stained gels.
NPM-ALK is expressed in normal T cells To study the influence of NPM-ALK in T cells of mice, the full-length cDNA of NPM-ALK fusion gene was cloned in vector under the control of the murine CD4 promoter (Figure 1A). Injection of this construct into blastocysts yielded 6 different NPM-ALK founders that were identified from 3 foster mothers. The copy number of the NPM-ALK transgene varied considerably among the different lines (Figure 1B). With the exception of one mouse (N8), all founders and their corresponding NPM-ALK progenies (N1, N14, N16) expressed the expected ALK fusion protein with a molecular weight of 80 kDa (Figure 1C). This protein corresponded to the NPM-ALK of human cell lines carrying the t(2;5) translocation and was expressed at levels similar to those of human ALCL-derived cell lines (Figure 1C). All 5 NPM-ALK-expressing founders were crossed to generate 5 different mouse lines. However, N5 and N15 died, before mating, of bilateral posterior limb paralysis and thymic tumor, respectively.
The CD4 transgene cassette allows the expression of the target protein
in all T cells, including early progenitor thymocytes (CD4+/CD8+) and single positive T cells
(CD4+/CD8 Stat3 and Jak3 are constitutively phosphorylated in NPM-ALK Tg mice Because NPM-ALK is constitutively autophosphorylated in human ALCL cells, we analyzed the phosphorylation status of NPM-ALK in transgenic cells and observed that in normal, as well as neoplastic NPM-ALK cells, it is constitutively phosphorylated (Figure 2A). Because activated ALK fusion proteins can efficiently bind Shc, PLC-, Grb-2, and
PI3K,29 we studied whether the transgenic NPM-ALK fusion
protein could efficiently bind the corresponding mouse proteins as
well. As shown in Figure 2B, mouse Shc, IRS-1, Grb-2, and PI3K proteins
efficiently bound NPM-ALK in normal as well as in neoplastic cells.
Moreover, we were able to demonstrate that phosphorylated Stat3 could
be coprecipitated with ALK (data not shown). Because NPM-ALK leads to
the constitutive activation of Stat3,21,22 and
Jak3,22 we further investigated the activation status of
these molecules in our NPM-ALK Tg mice. As shown in Figure 2C-D,
NPM-ALK Tg thymocytes, but not control cells, displayed constitutively
phosphorylated Stat3 and Jak3. Overall, these findings demonstrate that
the NPM-ALK transgene is constitutively activated in T cells and binds
to the same adaptor proteins as in humans. Thus, our transgenic model
mimics the molecular features of human NPM-ALK+
lymphomas.
Cellular phenotype and lymphoid organ development in NPM-ALK transgenic mice To characterize the putative effects resulting from the constitutive activation of NPM-ALK in T lymphocytes, we analyzed the morphologic and phenotypic features of T-cell lymphoid populations and their activation and differentiation states. Overall, the relative and absolute numbers of T and B lymphocytes, within primary and secondary lymphoid organs, were similar in Tg and control littermate mice. Microscopic evaluation demonstrated a normal lymphoid organization with the physiologic preservation of all lymphoid microenvironments. Finally, the histologic surveys of lung, kidney, stomach, intestine, testis, ovaries, and brain did not reveal any morphologic anomalies.Flow cytometry of NPM-ALK Tg thymocytes showed a normal distribution of
CD4
To determine whether the constitutive expression of NPM-ALK could possibly modify the survival and/or proliferative potential of T lymphocytes, NPM-ALK Tg thymocytes were incubated in vitro with different apoptotic stimuli. As shown in Figure 3C, both Tg and controls had similar rates of spontaneous and induced apoptosis. The in vitro proliferative rates of purified peripheral T lymphocytes, stimulated with suboptimal and to "ad hoc" concentrations of mitogens, were also similar in transgenic and control mice (Figure 3D). These findings indicate that NPM-ALK alone is not capable of significantly modifying the survival and cell growth of T lymphocytes from young mice in vitro. NPM-ALK transgenic mice develop spontaneous lymphoid tumors Mice from N1, N14, and N16 lines were healthy up to 5 to 7 weeks of life. After the fifth week, Tg animals started to develop tumors. Survival curves obtained from 86 mice for the N16 line and 110 mice for the N1 line showed a mean survival of 18.5 (Figure 4A), and 17 weeks (Figure 4B), respectively, with a overall incidence of 100% for both lines. Tumors were mainly represented by thymic lymphomas or plasma cell neoplasms (Figures 5 and 6), and all 3 lines developed, albeit with different frequencies, both thymic lymphomas and/or plasma cell tumors. Mice belonging to the N1 line showed, in fact, a prevalence of plasma cell tumors (> 80%), in contrast to N16 mice that more often developed thymic lymphomas (> 90%). Thymic and plasma cell tumors occurred with a similar frequency (50%) in mice of the N14 line. In rare cases (< 5% overall), we also found neoplasms characterized by atypical, spindle cells within a dense connective tissue. In addition, we also documented rare tumors (< 1%) characterized by immature cells with abundant cytoplasm lacking either T- or B-cell markers, but expressing CD11b. These tumors involved central and peripheral lymphoid tissues and were often observed infiltrating the liver, kidneys, lungs, and other internal organs.
The mediastinal T-cell lymphomas were composed of medium-sized
lymphoblasts, with a relatively high mitotic index (10-15 mitosis/10 high power field [hpf]) and high proliferation index as demonstrated by anti-Ki-67 staining (Figure 4C). These immature thymocytes were
always Thy-1+ and CD44+ but B220 Plasma cell tumors could be categorized into 3 major groups, based on
their cytologic features. The first group included tumors composed
primarily of mature plasma cells characterized by a large cytoplasm and
eccentric and sometime binucleated nuclei with evident nucleoli. The
second group included tumors with large, atypical cells with irregular
nuclei and conscious nucleoli. Finally, a subset of these neoplasms
displayed very atypical, pleomorphic/anaplastic cells (Figure 5A-D).
Plasmacytomas occurring in lymph nodes, spleen, and very rarely the
thymus often completely replaced these lymphoid organs and invariably
invaded the surrounding tissues. Furthermore, in a substantial subset
of the transgenic mice (20%), the neoplastic plasma cells occupied the
bone marrow spaces and invaded into the vertebral bones, compressing
and often destroying spinal ganglia and nerves (Figure 6A,B). In rare
instances the neoplastic cells, growing within the perispinal spaces,
even reached the central nervous system (Figure 6C). These histologic
findings corroborated the frequent gross limb paralysis of these mice
and other postural and behavioral (spinning and rotational) habits.
Notably, these plasma cell tumors occurred with the same frequency in
mice crossed in C57BL/6 and Balb/c backgrounds. Immunophenotypic
analysis of these neoplasms demonstrated that these tumors rarely
expressed B220/CD45R but were invariably NPM-ALK+ (Figure
6D) and CD138+ (Figure 6E). The proliferation rate as
measured by the Ki-67 staining was variable ranging from 10% to 40%
(Figure 6F). The B-cell origin of these tumors was further confirmed by
the Southern blotting (Figure 7A) and by
enzyme-linked immunosorbent assay (ELISA; data not shown). Furthermore,
immunohistochemical staining performed on paraffin-embedded tissue
samples demonstrated the clonotypic expression of heavy and light chain
of these tumors (Figure 6G). Moreover, free light chain immunoglobulin
was demonstrated in animals carrying plasma cell neoplasms (Figure
6H-I). Collectively, these findings demonstrate that these neoplasms
express clonal immunoglobulin, which can be secreted and detected in
the serum.
Finally, we investigated the expression profiles of several cell cycle
regulators and Stat3 and Stat5 in fresh tumor samples and in 3 NPM-ALK
T-cell lines. All NPM-ALK+ samples showed the constitutive
expression of phosphorylated Stat3 (Figure 7B). However, a single
NPM-ALK case displayed very low levels of phosphorylated Stat5, despite
the relatively high levels of Stat5. Interestingly the expressions of
c-myc, phospho-Erk-1/2, and cyclin A and D3 were similar in NPM-ALK and
in
We have produced and characterized a new mouse model of NPM-ALK-induced lymphomagenesis and have demonstrated that human NPM-ALK leads invariably to the generation of T-cell lymphomas and plasma cell tumors. Our findings show that ALK can efficiently bind a series of mouse adaptor proteins and result in the constitutive activation of Jak3 and Stat3. Nonetheless, in normal cells NPM-ALK alone does not increase cell proliferation and/or promote the survival of normal thymocytes to proapototic agents. Even though several groups have demonstrated the transforming potential of ALK chimeric proteins in vitro, and Kuefer et al18 have shown that NPM-ALK-containing retrovirus can lead to the transformation of hematopoietic cells toward B-cell large cell lymphomas, the ability of NPM-ALK to induce the transformation of T lymphocytes in vivo is still under debate. Indeed, understanding the pathogenetic role of NPM-ALK in T-cell transformation is important because most of the ALK+ human ALCLs are T cell in origin30 and ALK+ lymphocytes have been detected in healthy individuals.23 Our in vivo studies demonstrated that the constitutive activation of
ALK can successfully prompt, with a relatively short latency,
spontaneous lymphomagenesis in all mice. This is particularly interesting considering that in other murine transgenic models T-cell
tumors occur in only a subset of the animals.28,31,32 The
efficient ability of activated ALK to induce transformation may be due
to the diversity and complexity of the ALK-signaling pathway. In fact,
we and others have shown that PI3K, PLC- In our model the T-cell tumors were exclusively lymphoblastic lymphomas. The precise mechanism(s) for this prevalence is unclear. Because thymocytes proliferate rapidly, it is possible that cell proliferation may be an important requirement for the ALK-mediated transformation observed in our model. A relatively high rate of cell division may allow the acquisition of a sufficient number of genetic alterations capable of cooperating with ALK, thereby spurring transformation. Alternatively, the transformation of NPM-ALK thymocytes may occur because immature T cells express unique genes, which may promote the activation of unique pathways or facilitate genetic aberrations. For example, immature thymocytes actively undergo gene rearrangement of their TCR loci, and they are likely to undergo erroneous genetic recombination.33 These errors might lead to cell death but in the presence of ALK may fortuitously encourage or promote transformation. Immunophenotypic analysis showed that a fraction (approximately 50%) of ALK T cells coexpressed the CD30 antigen, a molecule that is invariably expressed by human ALCL.1 Because a very small subpopulation of normal thymocytes express detectable surface CD30,34 it is unclear whether the NPM-ALK CD30+ tumors may simply derive from these normal CD30+ thymocytes or whether CD30 could be up-regulated as a result of ALK expression or cell transformation. If NPM-ALK directly regulated the expression of CD30, one could predict the overexpression of CD30 in normal NPM-ALK Tg thymocytes. However, the analysis of normal transgenic cells did not show any CD30 up-regulation. Thus, transformed NPM-ALK+ cells may undergo nonspecific changes, such as chromatin remodeling, etc, which facilitate the transcription of other genes, including CD30, via transcription factors whose expression might be ALK mediated. These hypotheses require additional studies. Together with T-cell lymphomas, NPM-ALK Tg mice also developed ALK+ plasma cell tumors. The B-cell origin of these tumors was confirmed by the presence of specific heavy-chain immunoglobulin gene rearrangements and by expression of B-cell/plasma cell-associated antigens CD45R and CD138. Our results exclude the possibility that ALK tumors may develop as a result of the unique insertion of the transgene in proximity of genes expressed in B cells and/or responsible for the activation of target genes that facilitate B-cell transformation, because all the 3 transgenic lines developed such tumors. The constitutive expression of ALK in these neoplastic plasma cells in addition to their high incidence in NPM-ALK Tg mice strongly suggest that the occurrence of plasma cell neoplasms is due to the forced expression of ALK. Nevertheless, our CD4 cassette should allow the expression of the desired transgene only in T cells.24 The aberrant expression of ALK in the neoplastic plasma cells suggests that this cassette may be transcriptionally active in some B cells that may be committed to plasma cell differentiation and/or or in plasma cells. The expression of CD4 in normal B cells has not been described. However, the absence of the CD8 silencer in our construct may result in the inappropriate expression of the driven transgene. The low level of expression of CD30 in some non-T cells in CD30 Tg mice tends to support this hypothesis (data not shown). Alternatively, plasma cells may aberrantly transcribe CD4 and thus might have the appropriate transcription machinery to induce the expression of NPM-ALK in our transgenic B cells.35-37 Finally, Delsol et al16 have described a group of IgA+ plasmacytoid B cell tumors that overexpresses ALK and CD4 antigens.16 Regardless of the precise mechanisms leading to the aberrant expression of ALK in Tg plasma cells, NPM-ALK Tg mice are a suitable model to study plasma cell tumors and, in particular, multiple myeloma. In fact, in addition to peripheral plasma cell tumors, 20% of our Tg mice displayed primary neoplasms within the bone marrow, often involving the dorsal vertebrae. These tumors led to the compression and/or infiltration of ganglia and spinal nerves and ultimately resulted in the paralysis of the posterior legs. The clinical presentation and histologic features of these tumors closely recapitulated those of human multiple myelomas. Therefore, NPM-ALK transgenic mice represent the only murine model for multiple myeloma. In fact because the first study of Anderson et al,38 who demonstrated that after injection of pristane, Balb/c mice were prone to develop plasmacytomas, several investigators have described several other plasma cell models that do not display the features of human multiple meylomas.32,39-41 The discovery of rare plasmacytoid tumors overexpressing ALK in humans16 and the high frequency of plasma cell neoplasms in our mice strongly indicate that the forced expression of ALK could transactivate crucial pathways for the development of plasma cell dyscrasias. We and others have shown that ALK can constitutively transactivate Stat3. Stat3 is known to play an important role in the pathogenesis of multiple myelomas, and its activation is required for the maintenance and survival of neoplastic plasma cells.42 Interestingly, the activation of Stats might be achieved in multiple ways. This often occurs via the interleukin 6 receptor (IL-6R) engagement, but recently it was also shown that the inappropriate activation of fibroblast growth factor receptor 3 (FGFR3) can efficiently lead to Stat1 and Stat3 activation.43 Collectively, these findings strongly suggest that the transactivation of Stats, and in particular of Stat3, plays a crucial role in the pathogenesis of plasma cell tumors. Finally, activated ALK via Grb-2, Shc, and other adaptors leads to the activation of Ras and Erk1/2 (data not shown), and Ras has been demonstrated to have an important role in the pathogenesis of plasma cell neoplasms.44,45 In conclusion our findings have confirmed the tumorigenic activity of ALK in vivo and have shown that ALK can efficiently transform T lymphocytes and lead to the development of plasma cell neoplasms. Our model will provide a valuable tool to dissect the signaling of ALK and to identify new putative recurrent aberrations cooperating with ALK in promoting T-cell transformation. The NPM-ALK mice are the first in vivo murine model for multiple myeloma and represent a unique model in which to investigate the efficacy of new therapeutic approach for the treatment of both ALCL and multiple myelomas.
We thank Drs A. Rostagno and E. Zhu for their technical assistance.
Submitted May 30, 2002; accepted August 21, 2002.
Prepublished online as Blood First Edition Paper, November 7, 2002; DOI 10.1182/blood-2002-05-1343.
Supported by grant RO1-CA64033 from the National Institutes of Health and by an Associazione Italiana per la Ricerca sul Cancro (AIRC) grant.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Giorgio Inghirami, New York University, Department of Pathology and Kaplan Cancer Center, 550 First Ave, New York, NY 10016; e-mail: inghig01{at}med.nyu.edu.
1.
Stein H, Foss HD, Durkop H, et al.
CD30(+) anaplastic large cell lymphoma: a review of its histopathologic, genetic, and clinical features.
Blood.
2000;96:3681-3695 2. Rimokh R, Magaud JP, Berger F, et al. A translocation involving a specific breakpoint (q35) on chromosome 5 is characteristic of anaplastic large cell lymphoma (`Ki-1 lymphoma'). Br J Haematol. 1989;71:31-36[Medline] [Order article via Infotrieve].
3.
Kaneko Y, Frizzera G, Edamura S, et al.
A novel translocation, t(2;5)(p23;q35), in childhood phagocytic large T-cell lymphoma mimicking malignant histiocytosis.
Blood.
1989;73:806-813 4. Le Beau MM, Bitter MA, Larson RA, et al. The t(2;5)(p23;q35): a recurring chromosomal abnormality in Ki-1-positive anaplastic large cell lymphoma. Leukemia. 1989;3:866-870[Medline] [Order article via Infotrieve].
5.
Morris SW, Kirstein MN, Valentine MB, et al.
Fusion of a kinase gene, ALK, to a nucleolar protein gene, NPM, in non-Hodgkin's lymphoma.
Science.
1994;263:1281-1284 6. Wang D, Umekawa H, Olson MO. Expression and subcellular locations of two forms of nucleolar protein B23 in rat tissues and cells. Cell Mol Biol Res. 1993;39:33-42[Medline] [Order article via Infotrieve]. 7. Szebeni A, Olson MO. Nucleolar protein B23 has molecular chaperone activities. Protein Sci. 1999;8:905-912[Medline] [Order article via Infotrieve]. 8. Okuda M, Horn HF, Tarapore P, et al. Nucleophosmin/B23 is a target of CDK2/cyclin E in centrosome duplication. Cell. 2000;103:127-140[CrossRef][Medline] [Order article via Infotrieve]. 9. Iwahara T, Fujimoto J, Wen D, et al. Molecular characterization of ALK, a receptor tyrosine kinase expressed specifically in the nervous system. Oncogene. 1997;14:439-449[CrossRef][Medline] [Order article via Infotrieve]. 10. Morris SW, Naeve C, Mathew P, et al. ALK, the chromosome 2 gene locus altered by the t(2;5) in non-Hodgkin's lymphoma, encodes a novel neural receptor tyrosine kinase that is highly related to leukocyte tyrosine kinase (LTK). Oncogene. 1997;14:2175-2188[CrossRef][Medline] [Order article via Infotrieve]. 11. Duyster J, Bai RY, Morris SW. Translocations involving anaplastic lymphoma kinase (ALK). Oncogene. 2001;20:5623-5637[CrossRef][Medline] [Order article via Infotrieve].
12.
Souttou B, Carvalho NB, Raulais D, Vigny M.
Activation of anaplastic lymphoma kinase receptor tyrosine kinase induces neuronal differentiation through the mitogen-activated protein kinase pathway.
J Biol Chem.
2001;276:9526-9531
13.
Stoica GE, Kuo A, Aigner A, et al.
Identification of anaplastic lymphoma kinase as a receptor for the growth factor pleiotrophin.
J Biol Chem.
2001;276:16772-16779 14. Coffin CM, Patel A, Perkins S, Elenitoba-Johnson KS, Perlman E, Griffin CA. ALK1 and p80 expression and chromosomal rearrangements involving 2p23 in inflammatory myofibroblastic tumor. Mod Pathol. 2001;14:569-576[CrossRef][Medline] [Order article via Infotrieve].
15.
Lamant L, Pulford K, Bischof D, et al.
Expression of the ALK tyrosine kinase gene in neuroblastoma.
Am J Pathol.
2000;156:1711-1721
16.
Delsol G, Lamant L, Mariame B, et al.
A new subtype of large B-cell lymphoma expressing the ALK kinase and lacking the 2;5 translocation.
Blood.
1997;89:1483-1490
17.
Bai RY, Dieter P, Peschel C, Morris SW, Duyster J.
Nucleophosmin-anaplastic lymphoma kinase of large-cell anaplastic lymphoma is a constitutively active tyrosine kinase that utilizes phospholipase C-gamma to mediate its mitogenicity.
Mol Cell Biol.
1998;18:6951-6961
18.
Kuefer MU, Look AT, Pulford K, et al.
Retrovirus-mediated gene transfer of NPM-ALK causes lymphoid malignancy in mice.
Blood.
1997;90:2901-2910 19. Bischof D, Pulford K, Mason DY, Morris SW. Role of the nucleophosmin (NPM) portion of the non-Hodgkin's lymphoma-associated NPM-anaplastic lymphoma kinase fusion protein in oncogenesis. Mol Cell Biol. 1997;17:2312-2325[Abstract].
20.
Bai RY, Ouyang T, Miething C, Morris SW, Peschel C, Duyster J.
Nucleophosmin-anaplastic lymphoma kinase associated with anaplastic large-cell lymphoma activates the phosphatidylinositol 3-kinase/Akt antiapoptotic signaling pathway.
Blood.
2000;96:4319-4327
21.
Zhang Q, Raghunath PN, Xue L, et al.
Multilevel dysregulation of STAT3 activation in anaplastic lymphoma kinase-positive T/null-cell lymphoma.
J Immunol.
2002;168:466-474 22. Zamo A, Chiarle R, Piva R, et al. Anaplastic lymphoma kinase (ALK) activates Stat3 and protects hematopoietic cells from cell death. Oncogene. 2002;21:1038-1047[CrossRef][Medline] [Order article via Infotrieve].
23.
Maes B, Vanhentenrijk V, Wlodarska I, et al.
The NPM-ALK and the ATIC-ALK fusion genes can be detected in non-neoplastic cells.
Am J Pathol.
2001;158:2185-2193 24. Sawada S, Scarborough JD, Killeen N, Littman DR. A lineage-specific transcriptional silencer regulates CD4 gene expression during T lymphocyte development. Cell. 1994;77:917-929[CrossRef][Medline] [Order article via Infotrieve].
25.
Chiarle R, Podda A, Prolla G, Podack ER, Thorbecke GJ, Inghirami G.
CD30 overexpression enhances negative selection in the thymus and mediates programmed cell death via a Bcl-2-sensitive pathway.
J Immunol.
1999;163:194-205
26.
Piva R, Liu J, Chiarle R, Podda A, Pagano M, Inghirami G.
In Vivo interference with Skp1 function leads to genetic instability and neoplastic transformation.
Mol Cell Biol.
2002;22:8375-8387
27.
Chiarle R, Budel LM, Skolnik J, et al.
Increased proteasome degradation of cyclin-dependent kinase inhibitor p27 is associated with a decreased overall survival in mantle cell lymphoma.
Blood.
2000;95:619-626 28. Mangues R, Symmans WF, Lu S, Schwartz S, Pellicer A. Activated N-ras oncogene and N-ras proto-oncogene act through the same pathway for in vivo tumorigenesis. Oncogene. 1996;13:1053-1063[Medline] [Order article via Infotrieve].
29.
Slupianek A, Nieborowska-Skorska M, Hoser G, et al.
Role of phosphatidylinositol 3-kinase-Akt pathway in nucleophosmin/anaplastic lymphoma kinase-mediated lymphomagenesis.
Cancer Res.
2001;61:2194-2199 30. Kadin ME, Morris SW. The t(2;5) in human lymphomas. Leuk Lymphoma. 1998;29:249-256[Medline] [Order article via Infotrieve].
31.
Latres E, Chiarle R, Schulman BA, et al.
Role of the F-box protein Skp2 in lymphomagenesis.
Proc Natl Acad Sci U S A.
2001;98:2515-2520
32.
Suematsu S, Matsusaka T, Matsuda T, et al.
Generation of plasmacytomas with the chromosomal translocation t(12;15) in interleukin 6 transgenic mice.
Proc Natl Acad Sci U S A.
1992;89:232-235 33. Pasqualucci L, Neumeister P, Goossens T, et al. Hypermutation of multiple proto-oncogenes in B-cell diffuse large-cell lymphomas. Nature. 2001;412:341-346[CrossRef][Medline] [Order article via Infotrieve].
34.
Romagnani P, Annunziato F, Manetti R, et al.
High CD30 ligand expression by epithelial cells and Hassal's corpuscles in the medulla of human thymus.
Blood.
1998;91:3323-3332 35. Shimizu S, Takiguchi T, Fukutoku M, et al. Establishment of a CD4-positive plasmacytoma cell line (AMO1). Leukemia. 1993;7:274-280[Medline] [Order article via Infotrieve].
36.
Kubonishi I, Sonobe H, Miyagi T, et al.
A Ki-1 (CD30)-positive T (E+, CD4+, Ia+)-cell line, DL-40, established from aggressive large cell lymphoma.
Cancer Res.
1990;50:7682-7685 37. Spier CM, Grogan TM, Durie BG, et al. T-cell antigen-positive multiple myeloma. Mod Pathol. 1990;3:302-307[Medline] [Order article via Infotrieve]. 38. Anderson PN, Potter M. Induction of plasma cell tumours in BALB-c mice with 2,6,10,14-tetramethylpentadecane (pristane). Nature. 1969;222:994-995[CrossRef][Medline] [Order article via Infotrieve].
39.
Potter M, Wiener F.
Plasmacytomagenesis in mice: model of neoplastic development dependent upon chromosomal translocations.
Carcinogenesis.
1992;13:1681-1697 40. Suematsu S, Matsusaka T, Matsuda T, Hirano T, Kishimoto T. Interleukin-6 in myeloma/plasmacytoma. Int Rev Exp Pathol. 1993;34:91-98[Medline] [Order article via Infotrieve]. 41. Sugiyama H, Silva S, Wang YS, et al. Abelson murine leukemia virus transforms preneoplastic Emu-myc transgene-carrying cells of the B-lymphocyte lineage into plasmablastic tumors. Int J Cancer. 1990;46:845-852[Medline] [Order article via Infotrieve]. 42. Catlett-Falcone R, Landowski TH, Oshiro MM, et al. Constitutive activation of Stat3 signaling confers resistance to apoptosis in human U266 myeloma cells. Immunity. 1999;10:105-115[CrossRef][Medline] [Order article via Infotrieve]. 43. Hart KC, Robertson SC, Kanemitsu MY, Meyer AN, Tynan JA, Donoghue DJ. Transformation and Stat activation by derivatives of FGFR1, FGFR3, and FGFR4. Oncogene. 2000;19:3309-3320[CrossRef][Medline] [Order article via Infotrieve]. 44. Ozaki S, Kosaka M. Multiple myeloma: new aspects of biology and treatment. J Med Invest. 1998;44:127-136[Medline] [Order article via Infotrieve]. 45. Corradini P, Ladetto M, Inghirami G, Boccadoro M, Pileri A. N- and K-ras oncogenes in plasma cell dyscrasias. Leuk Lymphoma. 1994;15:17-20[Medline] [Order article via Infotrieve].
© 2003 by The American Society of Hematology.
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
|
 |
|
  Y. P. Mosse, A. Wood, and J. M. Maris Inhibition of ALK Signaling for Cancer Therapy Clin. Cancer Res., September 15, 2009; 15(18): 5609 - 5614. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Marzec, Q. Zhang, A. Goradia, P. N. Raghunath, X. Liu, M. Paessler, H. Y. Wang, M. Wysocka, M. Cheng, B. A. Ruggeri, et al. Oncogenic kinase NPM/ALK induces through STAT3 expression of immunosuppressive protein CD274 (PD-L1, B7-H1) PNAS, December 30, 2008; 105(52): 20852 - 20857. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Ambrogio, C. Voena, A. D. Manazza, C. Martinengo, C. Costa, T. Kirchhausen, E. Hirsch, G. Inghirami, and R. Chiarle The Anaplastic Lymphoma Kinase Controls Cell Shape and Growth of Anaplastic Large Cell Lymphoma through Cdc42 Activation Cancer Res., November 1, 2008; 68(21): 8899 - 8907. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Accornero, G. Lattanzio, T. Mangano, R. Chiarle, R. Taulli, F. Bersani, P. E. Forni, S. Miretti, C. Scuoppo, W. Dastru, et al. An In vivo Model of Met-Driven Lymphoma as a Tool to Explore the Therapeutic Potential of Met Inhibitors Clin. Cancer Res., April 1, 2008; 14(7): 2220 - 2226. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. G. Christensen, H. Y. Zou, M. E. Arango, Q. Li, J. H. Lee, S. R. McDonnell, S. Yamazaki, G. R. Alton, B. Mroczkowski, and G. Los Cytoreductive antitumor activity of PF-2341066, a novel inhibitor of anaplastic lymphoma kinase and c-Met, in experimental models of anaplastic large-cell lymphoma Mol. Cancer Ther., December 1, 2007; 6(12): 3314 - 3322. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. B. Staber, P. Vesely, N. Haq, R. G. Ott, K. Funato, I. Bambach, C. Fuchs, S. Schauer, W. Linkesch, A. Hrzenjak, et al. The oncoprotein NPM-ALK of anaplastic large-cell lymphoma induces JUNB transcription via ERK1/2 and JunB translation via mTOR signaling Blood, November 1, 2007; 110(9): 3374 - 3383. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. M. Amin and R. Lai Pathobiology of ALK+ anaplastic large-cell lymphoma Blood, October 1, 2007; 110(7): 2259 - 2267. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Galietta, R. H. Gunby, S. Redaelli, P. Stano, C. Carniti, A. Bachi, P. W. Tucker, C. J. Tartari, C.-J. Huang, E. Colombo, et al. NPM/ALK binds and phosphorylates the RNA/DNA-binding protein PSF in anaplastic large-cell lymphoma Blood, October 1, 2007; 110(7): 2600 - 2609. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Leventaki, E. Drakos, L. J. Medeiros, M. S. Lim, K. S. Elenitoba-Johnson, F. X. Claret, and G. Z. Rassidakis NPM-ALK oncogenic kinase promotes cell-cycle progression through activation of JNK/cJun signaling in anaplastic large-cell lymphoma Blood, September 1, 2007; 110(5): 1621 - 1630. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Liu, Z. Liu, S.-W. Jang, Z. Ma, K. Shinmura, S. Kang, S. Dong, J. Chen, K. Fukasawa, and K. Ye Sumoylation of nucleophosmin/B23 regulates its subcellular localization, mediating cell proliferation and survival PNAS, June 5, 2007; 104(23): 9679 - 9684. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Li, D. P. Sejas, S. Burma, D. J. Chen, and Q. Pang Nucleophosmin suppresses oncogene-induced apoptosis and senescence and enhances oncogenic cooperation in cells with genomic instability Carcinogenesis, June 1, 2007; 28(6): 1163 - 1170. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Voena, C. Conte, C. Ambrogio, E. Boeri Erba, F. Boccalatte, S. Mohammed, O. N. Jensen, G. Palestro, G. Inghirami, and R. Chiarle The Tyrosine Phosphatase Shp2 Interacts with NPM-ALK and Regulates Anaplastic Lymphoma Cell Growth and Migration Cancer Res., May 1, 2007; 67(9): 4278 - 4286. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. V. Galkin, J. S. Melnick, S. Kim, T. L. Hood, N. Li, L. Li, G. Xia, R. Steensma, G. Chopiuk, J. Jiang, et al. Identification of NVP-TAE684, a potent, selective, and efficacious inhibitor of NPM-ALK PNAS, January 2, 2007; 104(1): 270 - 275. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Fawal, F. Armstrong, S. Ollier, H. Dupont, C. Touriol, B. Monsarrat, G. Delsol, B. Payrastre, and D. Morello A "liaison dangereuse" between AUF1/hnRNPD and the oncogenic tyrosine kinase NPM-ALK Blood, October 15, 2006; 108(8): 2780 - 2788. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Qiu, R. Lai, Q. Lin, E. Lau, D. M. Thomazy, D. Calame, R. J. Ford, L. W. Kwak, R. A. Kirken, and H. M. Amin Autocrine release of interleukin-9 promotes Jak3-dependent survival of ALK+ anaplastic large-cell lymphoma cells Blood, October 1, 2006; 108(7): 2407 - 2415. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Quintanilla-Martinez, S. Pittaluga, C. Miething, M. Klier, M. Rudelius, T. Davies-Hill, N. Anastasov, A. Martinez, A. Vivero, J. Duyster, et al. NPM-ALK-dependent expression of the transcription factor CCAAT/enhancer binding protein beta in ALK-positive anaplastic large cell lymphoma Blood, September 15, 2006; 108(6): 2029 - 2036. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Jacobsen Anaplastic Large-Cell Lymphoma, T-/Null-Cell Type Oncologist, July 1, 2006; 11(7): 831 - 840. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kasprzycka, M. Marzec, X. Liu, Q. Zhang, and M. A. Wasik From the Cover: Nucleophosmin/anaplastic lymphoma kinase (NPM/ALK) oncoprotein induces the T regulatory cell phenotype by activating STAT3 PNAS, June 27, 2006; 103(26): 9964 - 9969. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Li, D. P. Sejas, R. Rani, T. Koretsky, G. C. Bagby, and Q. Pang Nucleophosmin Regulates Cell Cycle Progression and Stress Response in Hematopoietic Stem/Progenitor Cells J. Biol. Chem., June 16, 2006; 281(24): 16536 - 16545. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Piva, R. Chiarle, A. D. Manazza, R. Taulli, W. Simmons, C. Ambrogio, V. D'Escamard, E. Pellegrino, C. Ponzetto, G. Palestro, et al. Ablation of oncogenic ALK is a viable therapeutic approach for anaplastic large-cell lymphomas Blood, January 15, 2006; 107(2): 689 - 697. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Mathas, K. Johrens, S. Joos, A. Lietz, F. Hummel, M. Janz, F. Jundt, I. Anagnostopoulos, K. Bommert, P. Lichter, et al. Elevated NF-{kappa}B p50 complex formation and Bcl-3 expression in classical Hodgkin, anaplastic large-cell, and other peripheral T-cell lymphomas Blood, December 15, 2005; 106(13): 4287 - 4293. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Ambrogio, C. Voena, A. D. Manazza, R. Piva, L. Riera, L. Barberis, C. Costa, G. Tarone, P. Defilippi, E. Hirsch, et al. p130Cas mediates the transforming properties of the anaplastic lymphoma kinase Blood, December 1, 2005; 106(12): 3907 - 3916. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Bacchiocchi, G. Baldanzi, D. Carbonari, C. Capomagi, E. Colombo, W. J. van Blitterswijk, A. Graziani, and F. Fazioli Activation of {alpha}-diacylglycerol kinase is critical for the mitogenic properties of anaplastic lymphoma kinase Blood, September 15, 2005; 106(6): 2175 - 2182. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Korgaonkar, J. Hagen, V. Tompkins, A. A. Frazier, C. Allamargot, F. W. Quelle, and D. E. Quelle Nucleophosmin (B23) Targets ARF to Nucleoli and Inhibits Its Function Mol. Cell. Biol., February 15, 2005; 25(4): 1258 - 1271. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Li, X. Zhang, D. P. Sejas, G. C. Bagby, and Q. Pang Hypoxia-induced Nucleophosmin Protects Cell Death through Inhibition of p53 J. Biol. Chem., October 1, 2004; 279(40): 41275 - 41279. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T.-L. Gu, Z. Tothova, B. Scheijen, J. D. Griffin, D. G. Gilliland, and D. W. Sternberg NPM-ALK fusion kinase of anaplastic large-cell lymphoma regulates survival and proliferative signaling through modulation of FOXO3a Blood, June 15, 2004; 103(12): 4622 - 4629. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Bonvini, H. D. Rosa, N. Vignes, and A. Rosolen Ubiquitination and Proteasomal Degradation of Nucleophosmin-Anaplastic Lymphoma Kinase Induced by 17-Allylamino-Demethoxygeldanamycin: Role of the Co-Chaperone Carboxyl Heat Shock Protein 70-Interacting Protein Cancer Res., May 1, 2004; 64(9): 3256 - 3264. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. D. Gascoyne, L. Lamant, J. I. Martin-Subero, V. S. Lestou, N. L. Harris, H.-K. Muller-Hermelink, J. F. Seymour, L. J. Campbell, D. E. Horsman, I. Auvigne, et al. ALK-positive diffuse large B-cell lymphoma is associated with Clathrin-ALK rearrangements: report of 6 cases Blood, October 1, 2003; 102(7): 2568 - 2573. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. De Paepe, M. Baens, H. van Krieken, B. Verhasselt, M. Stul, A. Simons, B. Poppe, G. Laureys, P. Brons, P. Vandenberghe, et al. ALK activation by the CLTC-ALK fusion is a recurrent event in large B-cell lymphoma Blood, October 1, 2003; 102(7): 2638 - 2641. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Onciu, F. G. Behm, J. R. Downing, S. A. Shurtleff, S. C. Raimondi, Z. Ma, S. W. Morris, W. Kennedy, S. C. Jones, and J. T. Sandlund ALK-positive plasmablastic B-cell lymphoma with expression of the NPM-ALK fusion transcript: report of 2 cases Blood, October 1, 2003; 102(7): 2642 - 2644. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
 |
 |
|||||||
 |
 |
 |
 |
 |
 |
 |
 |
||
| Copyright © 2003 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
Top
Abstract
Introduction
/
cul1 Tg mice were obtained by placing
(SacI/SalI) the human Cul1-N252 cDNA (encoding 1 to 252 [N252] amino terminal residues) into the CD4 cassette
Tg.
