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Leukemia & Lymphoma

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Leptin Receptor and Leukemia

a a a a
Masayuki Hino , Takafumi Nakao , Takahisa Yamane , Kensuke Ohta , Takayuki
a a
Takubo & Noriyuki Tatsumi
a
Department of Clinical Hematology, Osaka dry University Medical School, 1-5-7,
Asahimachi, Abeno-ku, Osaka, 545-8586, Japan
Published online: 01 Jun 2015.

To cite this article: Masayuki Hino, Takafumi Nakao, Takahisa Yamane, Kensuke Ohta, Takayuki Takubo & Noriyuki
Tatsumi (2000) Leptin Receptor and Leukemia, Leukemia & Lymphoma, 36:5-6, 457-461

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 NV
0Z(WN OPA IOvcnra* Puhli\hen A\~r~ciatinnl
Puhli\ht.d hy liuenre under
the Harwood Acadeniic Puhli\her* imprint.
pan oi the Gordon and Breach Puhli\hing Group.
Printed in Mdhy\i.i

Leptin Receptor and Leukemia

MASAYUKI HINO*. TAKAFUMI NAKAO, TAKAHISA YAMANE, KENSUKE OHTA, TAKAYUKI TAKUBO and
NORIYUKI TATSUMI

The rcccptor for Ieptin, the gene product of the obese gene, is expressed in hematopoietic
stem cells. Leptin stimulates normal myeloid and erythroid development, and is secreted from
bone marrow adipocytes, which occupy most of the marrow cavity in humans. Leptin might
Downloaded by [] at 03:10 18 July 2015

thus play an important role in the control of the expansion and differentiation of primitive
hematopoietic cells through paracrine interaction in the bone marrow microenvironment.
Leukemic cells of some patients with acute myeloblastic leukemia, acute lymphoblastic leu-
kemia, and chronic myeloid leukemia (CML) also express the leptin receptor. In cases of
CML. higher expression of leptin receptor is observed during blast crisis than in chronic
phase. Leptin alone and in combination with other cytokines has stimulative effects on prolif-
eration of leukemia cells as well as anti-apoptotic effects. These findings suggest the possibil-
ity that leptin plays roles in the pathophysiology of leukemia.

Keyitwi/.vc Leptin receptor. hematopoiesis. leukemia

1. INTRODUCTION 2. LEPTIN AND ITS RECEPTOR

Hematopoiesis is regulated by a large number of Leptin is a 16k Da protein produced by adipocytes4*',

cytokines, and is mediated through high-affinity bind-
ing of cytokines to specific cell-surface receptors.
Recently the receptor for leptin, the product of the
obese (ob) gene', was detected in hematopoietic pro-
genitor cells. Leptin stimulates myeloid and erythroid
In addition to functioning in normal
hematopoietic cells, several cytokines and these
receptors are involved in the survival, proliferation,
and differentiation of leukemic cells. It is thus impor-
tant to determine the expression of leptin receptor in
leukemic cells and the effects of leptin on such cells.
* Correspondence: Masayuki Hino, Department of Clinical Haernatology, Osaka City University Medical School 1-5-7, Aaahimachi.
Abeno-ku, Osaka 545-8586 Japan
and regulates nutrient intake and r n e t a b o l i ~ m ~ * ~ * ~ - ~ .
The severe obese phenotype in obhb mice is caused
by the inability to produce leptin, and daily adminis-
tration of recombinant leptin reduces body weight,
body fat, and food intake and increases metabolic rate
and physiological a ~ t i v i t y " ~ ' ~
Tartaglia
'~. et a]. have
isolated cDNAs encoding murine and human leptin
receptors". Leptin receptor is a single mem-
brane-spanning receptor, and exhibits sequence simi-
larity with the gp 130 signal-transducing component
of the interleukin (1L)-6 receptor' I , granulocyte col-
ony-stimulating factor (G-CSF) receptor", and leuke-
mia inhibitory factor (LIF) receptorI3. The leptin

457
 458 MASAYUKl HINO et al.
receptor is a member of the hematopoietic cytokine
receptor super-family which is characterized by a 200
amino acid conserved domain that includes four posi-
tionally conserved cysteine residues and a
Trp-Ser-X-Trp-Ser (WSXWS) motif. The leptin
receptor has several alternatively spliced forms (long
form and short form) with cytoplasmic domains of
different length^'^^'^*'^*'^. The long isoform of leptin
receptor is preferentially expressed in the hypothala-
mus of wild-type mice. It can activate signal transduc-
ers and activators of transcription (STAT)-3, STAT-5,
and STAT-6I4. In db/db mice, a point mutation within
this long isoform of the leptin receptor gene generates
a truncated shorter isoform of leptin receptor by
abnormal splicing, and expression of the long isoform
of leptin receptor is thereby suppressed". In contrast,
a short isoform of leptin receptor is present in both
Downloaded by [] at 03:10 18 July 2015
db/db mice and wild-type mice. The isoforms with a
shorter cytoplasmic domain are unable to activate the
STAT pathway, resulting in the db/db p h e n ~ t y p e ' ~ , ' ~ . cell-to-cell contact between hematopoietic progenitor
3. LEPTIN AND HEMATOPOIESIS
The proliferation and differentiation of hematopoietic
cells is regulated by various hematopoietic cytokines.
Hematopoietic cytokines exert biological effects
through specific receptors on the surface of hemat-
opoietic stem cells. Recently, the murine and human
leptin receptors have been cloned from murine yolk
sac-derived cell linesi6 and human CD34-positive
hematopoietic stem cells2'. As shown in Figure 2,
leptin receptor is expressed in isolated CD34-positive
hematopoietic stem cells and a variety of hematopoi-
etic cell lines2i9'6+22.It has been reported that human
CD34-positive hematopoietic stem cells and
CD 19/CD20-positive B lymphocytes express both
long and short forms of the leptin receptor2'. Several
studies have shown that Leptin can induce prolifera-
tion, differentiation, and functional activation of
hematopoietic cells in vitro. Leptin enhanced
cytokine production and phagocytosis of Leishmania
parasites by murine peritoneal macrophages2. Leptin
had stimulatory effects on myeloid, lymphoid, and
erythroid colony formation. Leptin acted synergisti-
cally with stem cell factor (SCF) in promoting prolif-
eration of primitive hematopoietic progenitors in
vitro3. Leptin alone increased numbers of macro-
phage and granulocyte colonies23. In the db/db mice,
which has a truncated non-functional leptin receptor,
steady-state levels of peripheral B cells and
CDCexpressing T cells are dramatically reduced2'.
The myeloid and lymphoid colony-forming potential
of the db/db mice bone marrow cells are also reduced
compared to that of wild-type control bone marrow
cells. Moreover, despite normal peripheral blood red
cell counts, spleen erythrocyte production is decreased
in these mice. These findings suggest that leptin has a
direct effect on early hematopoietic development. Fur-
thermore, several studies have found a significant cor-
relation of serum leptin level with blood cell count,
independent of body mass index and serum insulin
Constitutive hematopoiesis is regulated in
the bone marrow microenvironment by direct
cells and bone marrow stromal cells that include
fibroblasts, endothelial cells, macrophages, and adi-
pocytes. Leptin is expressed and secreted at high levels
during bone marrow adipocyte differentiation26.This is
relevant to hematopoiesis, since bone marrow adi-
pocytes occupy most of the marrow cavity in humans.
Leptin could play an important role in the control of the
expansion and differentiation of primitive hematopoi-
etic cells through paracrine interaction in the bone mar-
row microenvironment (Figure l). It has recently been
reported that leptin is present in human cord blood.
These findings suggest that leptin may play a role in
human fetal hematop~iesis~~.
4. EXPRESSION OF LEPTIN RECEPTOR IN
LEUKEMIA
In addition to normal hematopoietic cells, leukemic
cells have been shown to express functional receptors
for a variety of cytokines such as IL-3, IL-6, granulo-
cyte-macrophage colony stimulating factor (GM-CSF),
G-CSF, macrophage colony stimulating factor
(M-CSF), SCF, and thrombopoietin
These cytokines also regulate the survival, prolifera-
 LEPTIN RECEPTOR & LEUKEMIA 45’)

hematopoietic progenitor cells than in mature neutro-

stem cell

adipocyte
@

microenvironment
6‘
hematopoiesis

leptin

\
proliferation
differentiation
phils*-*’. Konopleva et a1 found that CD34TD33+
cells and CD34TD13’ cells (promyelocytes) did not
have the long isoform and expressed only very low lev-
els of the short isoform**. These results suggest rapid
down-regulation of leptin receptor expression during
myeloid differentiation.

5. BIOLOGICAL EFFECTS OF LEPTIN ON

leukemic cell proliferation LEUKEMIC CELLS

FIGURE I Model of physiological and pathophysiological roles of Leptin stimulates proliferation of the human mye-
leptin in hernatopoiesis and leukemia
loid leukemia cell lines OCI/AML2 and M 0 7 E in a
dose-dependent mannerz2. However, the proliferative
responses of these cell lines were not correlated with
tion, and differentiation of leukemic cells. Disregulated
the level of leptin receptor expression. Leptin induces
Downloaded by [] at 03:10 18 July 2015

expression of cytokines andor their receptors could be

proliferation of primary leukemic cells from some
involved in the pathogenesis of leukemia. Leptin recep-
AML patients. As shown in Figure 3, the combination
tor mRNA has been detected in several myeloid and
of leptin and GM-CSF synergistically increased the
lymphoid leukemic cell lines16q20321. Overexpression
number of leukemic cells from a patient with CML
of leptin receptor was observed in K562, HEL, and
(in blast crisis). Konopleva et al have reported that
M07E cells”. As shown in Figure 2, leptin receptors
freshjy prepared leukemic cells from 3 of 14 AML
were constitutively expressed in leukemic cells in some
patients exhibited a proliferative response to leptin
newly diagnosed cases of AML, ALL, and CML”.”,
alone, and that combinations of leptin with other
but were not detected in cases of chronic lymphocytic
hematopoietic cytokines (IL-3, G-CSF, and SCF)
leukemia”. Konopleva et al. found that long isoform,
induced an additively or a synergistically proliferative
short isoform, and both isoforms of leptin receptor
response in 7 of 14 AML cases2?. Furthermore, leptin
were expressed in 57.776, 59.1%~~ and 45.4% of cases
increased the number of AML progenitor cells (col-
of AML, respectively”. The incidence of leptin recep-
ony-forming cells)**. There is no evidence that leptin
tor expression was higher in recurrent cases of AML
can up-regulate the expression of other cytokine
than in newly diagnosed cases22. Expression of the
receptors. On the other hand, leptin prevented apopto-
long, but not the short, isoform occurs significantly
sis induced by cytokine withdrawal in
more often in primary AML than in secondary AML or
cytokine-dependent M 0 7 E and TF- 1 cells”. Various
myelodysplastic syndrome. No correlation was found
cytokines prevent apoptosis, and thereby promote cell
between leptin receptor expression and FAB classifica-
survival. Bone marrow contains many adipocytes,
tion, cytogenetic parameters, blast count, or response to
which create a high local concentration of leptin
In cases of CML (Figure 2 ) , higher
within the bone marrow microenvironment. Estey et
expression of leptin receptor was observed during blast
al. reported observing an increase in BMI in patients
crisis than in chronic phase’’. Expression of leptin
with acute promyelocytic leukemia. These findings
receptor was decreased during in vitro differentiation
could have implications for the pathophysiology of
of leukemic cells”. It thus appeared that expression of
leukemia. The biological and pathological roles of
the leptin receptor was associated with immature leu-
leptin in hematopoiesis and leukemogenesis will be
kemic cells. Similarly, higher expression of leptin
clarified by further investigations.
receptor was observed in CD34-positive immature
 460 MASAYUKI HlNO er a\.
FIGURE 2 Expression of the leptin receptor in fresh leukemic cells
Total cellular RNAs (I0 pg) prepared from cell lines or leukemic
cells of patients with AML, ALL, or CML during chronic phase
(CP) or in blast crisis (BC) were electrophoresed, transferred, and
hybridized with the human leptin receptor cDNA and rat GAPDH
cDNA. The various lanes contained RNAs extracted from the indi-
cated cell lines and patients. 1: K562, 2: HEL, 3: M07E. 4:
MOLT4, 5: CCRF-CEM, 6: isolated CD34-positive cells, 7: bone
marrow cells. 8: AMLI(M2). 9: AML2(M3), 10: AML3(M6), I I :
AML4(M6), 12: ALLI, 13: CMLl(CP), 14: CMLZ(CP). 15:
CML3(BC), 16: CMIA(BC)
Downloaded by [] at 03:10 18 July 2015
I and Friedman, J. M. (1995) Weight-reducing effects of the
0.301, F .- . I IL-6 signal transducer, gpl30. Cell, 63, 1149-1 157.
0 l.OPg/ml 1.O ng/ml 1.0 pgg/ml
Lepbn
FIGURE 3 Effects of leptin on proliferation of leukemic cells Leu-
kemic cells (1x106/ml) prepared from a patient with CML in blast
crisis were cultivated with increasing concentrations of recombi-
nant human leptin (provided by Amgen, Inc.) alone (open circles)
or in combination with 5 ng/ml GM-CSF (closed circles). The pro-
liferation of cells was determined by M l T assay. Results are
expressed as means i SD of six cultures
Acknowledgements
This work was supported in part by a Grant-in-aid
from the Ministry of Education, Science, and Culture
of Japan. We thank Amgen, Inc. (Thousand Oaks,
CA) for providing recombinant human leptin.
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