Abstracts

delivery. SUV39H1 knock down inhibits H3K9me3, growth of

myeloma cells, induces apoptosis, cell cycle deregulation and spontaneous accumulation of DNA double strand breaks. According to these
results, SUV39H1 depletion sensitizes myeloma cells to melphalan.
Chaetocin is a selective inhibitor of SUV39H1. We identied that
chaetocin has anti-myeloma effects, at low nanomolar doses, on 11
different myeloma cell lines, that are representative of the molecular
heterogeneity of the patients, in association with H3K9 trimethylation
inhibition. Furthermore, this signicant toxicity of chaetocin in MM
was conrmed on primary myeloma cells of 5 patients cocultured with
their bone marrow microenvironment without signicant toxicity on
normal bone marrow cells and hematopoietic stem cells. Interestingly,
the IC50 doses of chaetocin in MM were 50 fold lower compared to
results published in AML, suggesting H3K9 HMTs could be a potent
therapeutic target in MM.

Moreover, GEP performed on inducible Tet-shJunB MM.1S cells cocultured with BMSCs suggested a key role for JunB in the regulation
of Mcl-1 and c-Myc expression. Furthermore, knockdown of JunB
overcame resistance of MM cells to dexamethasone. Conversely, 4OHT treatment of MM cell lines transduced with pMSCV-JunBER-IRES-GFP but not pMSCV-IRES-GFP induced signicant
JunB/AP-1 luciferase activity and protected MM cells against bortezomib-induced apoptosis and ER stress. Ongoing experiments aim to
conrm the in vivo relevance of our in vitro data in a MM xenograft
mouse model inoculated with inducible Tet-shJunB MM.1S cells.
Conclusion: Taken together, our data demonstrate for the rst time
an important and surprising role of JunB/AP-1 in MM tumorigenesis
and strongly propose it as a novel therapeutic target in MM.

PO-233
PO-232
The AP-1 Transcription Factor JunB
Promotes Multiple Myeloma (MM) Cell
Proliferation, Survival and Drug Resistance
in the Bone Marrow Microenvironment
F. Fan,1 S. Vallet,1 M. Sattler,2 G. Tonon,3
M.H. Bashari,1 L. Bakiri,4 M. Jarahian,5 A. Roccaro,2
I. Ghobrial,2 C. Ball,5 H. Glimm,5 K.C. Anderson,2
H. Goldschmidt,5 E.F. Wagner,4 D. Jaeger,1 K. Podar1
1

Medical Oncology, National Center for Tumor Diseases (NCT), Uni2

versity of Heidelberg, Heidelberg, Germany; Department of Medical

Oncology, Dana-Farber Cancer Institute, Boston, MA; 3San Raffaele
Scientic Institute, Milan, Italy; 4Genes Development and Disease
Group, Spanish National Cancer Research Centre, Madrid, Spain;
5

National Center for Tumor Diseases (NCT), University of Heidelberg,

Heidelberg, Germany

Introduction: The family of activator protein-1 (AP-1) transcription factors has been implicated in a multitude of physiologic processes,
but also tumorigenesis. In multiple myeloma (MM), the role of AP-1 is
largely unknown. Materials and Methods: MM cells were cocultured with primary bone marrow stromal cells (BMSCs) or BMSC
lines. AP-1 expression was measured by western blot analysis and
qPCR. To delineate the specic functional role of JunB in MM
pathogenesis, we used knockdown and overexpression approaches
followed by 3H-thymidine incorporation, ow cytometry and western blot analysis, as well as gene expression proling (GEP), and a
MM xenograft mouse model. Results: Surprisingly, co-cultures of
MM cells with BMSCs rapidly and strongly induced sustained
expression of JunB, but not of other AP-1 family members. Induction
of JunB is predominantly mediated by soluble factors (i.e IL-6)
secreted by BMSCs rather than direct MM-BMSC contact. Pharmacologic inhibition identied the requirement of MEK/ERK and
NF-kB for BMSC-induced JunB expression and AP-1 transcriptional
activity. Functionally, signicant inhibition of proliferation was
observed in MM cells carrying pLKO.1-JunB shRNA, but not
pLKO.1-scrambled shRNA. In contrast, knockdown of other AP-1
family members had minor effects on MM cell proliferation.

A novel function of DEPTOR in

multiple myeloma: commitment to
plasma cell maturation
D. Quwaider, A.B. Herrero, P. Krzeminski,
L.A. Corchete, I. Misiewicz-Krzeminska,
M.E. Sarasquete, J.J. Prez, N. Puig, R. Garca-Sanz,
N.C. Gutirrez
Hematology Department, University Hospital, IBSAL IBMCC (USALCSIC), Salamanca, Spain

Funding: Asociacin Espaola Contra el Cncer (AECC:

GCB120981SAN). Background: The understanding of plasma cell
(PC) development could provide new insights for therapeutic strategies in MM. Here we explore the role of DEPTOR in the differentiation of B lymphocytes (BL) to PCs and its potential function
in myeloma pathogenesis. Materials and methods: H929 and
MM1S cell lines, and 10 healthy controls of bone marrow were
included in the study. Cell sorting was used to isolate different B cell
populations. mRNA expression was determined using microarrays
(Human gene 2.0 ST array) and qRT-PCR, and protein levels were
assessed by WB. Knockdown experiments were carried out using
commercial siRNAs. Results: Previously published gene expression
analysis by microarrays showed that DEPTOR was overexpressed in
normal PC compared to normal BL (GSE6691 at GEO repository).
This nding was conrmed by qRT-PCR. Thus, DEPTOR was
signicantly overexpressed in PC (mean 2- CT, 0.05940.0574)
compared to immature B cells (0.000900.00089), nave B cells
(0.0012480.0011) and memory B cells (0.001450.00098)
(P 0.001). To investigate the putative functional role of DEPTOR
in MM, we carried out loss-of-function experiments in H929 and
MM1S cell lines. Gene expression proling in H929 revealed
decreased levels of CD38 and IRF4, and overexpression of PAX5 in
DEPTOR-knockdown cells compared to control (q<0.05). Similar
results were obtained by using qRT-PCR and Western blot.
Knockdown of DEPTOR also resulted in an evident reduction in
cell size and loss of endoplasmic reticulum expansion, further supporting the reversion of plasma cell state into pre-plasmablast.
High levels of DEPTOR have been reported in some MM, especially
in those carrying c-MAF/MAFB translocations. Here, we also investigated whether DEPTOR expression could be affected by deregulation

15th International Myeloma Workshop, September 23-26, 2015

- e215