Dual NAMPT and BTK Targeting Leads to Synergistic Killing of Waldenström Macroglobulinemia Cells Regardless of MYD88 and CXCR4 Somatic Mutation Status.
OBJECTIVE
Nicotinamide phosphoribosyltransferase (Nampt) regulates intracellular NAD+pool and is highly expressed in a number of malignancies. FK866, a selective inhibitor of Nampt, depletes intracellular NAD+levels, thereby blocking cellular metabolism and triggering sensitization to other drugs and cell death. Here we characterized the antitumor effects of Nampt inhibition in Waldenström macroglobulinemia.


METHODS
We investigated Nampt role in MW cells using both mRNA and protein expression analyses. We have also used loss-of-function approaches to investigate the growth and survival effects of Nampt on MW cells and further tested the anti-MW activity of dual Nampt and BTK inhibition in vitro and in vivo RESULTS: We found that Waldenström macroglobulinemia cells exhibit high levels of Nampt compared with normal B cells. Loss of function studies suggested a potential oncogenic role of Nampt in Waldenström macroglobulinemia cells, and BTK-inhibitor ibrutinib and FK866 resulted in a significant and synergistic anti-Waldenström macroglobulinemia cell death, regardless of MYD88 and CXCR4 mutational status. Cell death was associated with: (i) activation of caspase-3, PARP and downregulation of Mcl-1, (ii) enhanced intracellular ATP and NAD+depletion, (iii) inhibition of NF-κB signaling, and (iv) inhibition of multiple prosurvival signaling pathways. In a murine xenograft Waldenström macroglobulinemia model, low-dose combination FK866 and ibrutinib is well tolerated, significantly inhibits tumor growth, and prolongs host survival.


CONCLUSIONS
Our results show intracellular NAD+level as crucial for proliferation and survival of Waldenström macroglobulinemia cells, and provides the mechanistic preclinical rationale for targeting Nampt, either alone or with Ibrutinib, to overcome drug resistance and improve patient outcome in Waldenström macroglobulinemia. Clin Cancer Res; 22(24); 6099-109. ©2016 AACR.
LeBow Institute for Myeloma Therapeutics and Jerome Lipper Center for Multiple Myeloma Research, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts. michele.cea@unige.it.|Department of Medicine (DiMI), University of Genoa, AOU, I.R.C.C.S. San Martino-IST, Genova, Italy.
Michele
M
Cea
LeBow Institute for Myeloma Therapeutics and Jerome Lipper Center for Multiple Myeloma Research, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.|Department of Medicine (DiMI), University of Genoa, AOU, I.R.C.C.S. San Martino-IST, Genova, Italy.
Antonia
A
Cagnetta
LeBow Institute for Myeloma Therapeutics and Jerome Lipper Center for Multiple Myeloma Research, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
Chirag
C
Acharya
Mount Auburn Hospital, Harvard Medical School, Cambridge, Massachusetts.
Prakrati
P
Acharya
LeBow Institute for Myeloma Therapeutics and Jerome Lipper Center for Multiple Myeloma Research, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
Yu-Tzu
YT
Tai
Bing Center for Waldenstrom's Macroglobulinemia, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
Cao
C
Yang
Department of Medicine (DiMI), University of Genoa, AOU, I.R.C.C.S. San Martino-IST, Genova, Italy.
Davide
D
Lovera
Department of Medicine (DiMI), University of Genoa, AOU, I.R.C.C.S. San Martino-IST, Genova, Italy.
Debora
D
Soncini
Department of Medicine (DiMI), University of Genoa, AOU, I.R.C.C.S. San Martino-IST, Genova, Italy.
Maurizio
M
Miglino
Laboratories Department, Pathology Unit, IRCCS AUO S. Martino-IST, Genova, Italy.
Giulio
G
Fraternali-Orcioni
Department of Surgical and Diagnostic Sciences (DISC), Pathology Unit, University of Genoa and IRCCS AUO S. Martino-IST, Genova, Italy.
Luca
L
Mastracci
Department of Medicine (DiMI), University of Genoa, AOU, I.R.C.C.S. San Martino-IST, Genova, Italy.
Alessio
A
Nencioni
Department of Medicine (DiMI), University of Genoa, AOU, I.R.C.C.S. San Martino-IST, Genova, Italy.
Fabrizio
F
Montecucco
Department of Medicine (DiMI), University of Genoa, AOU, I.R.C.C.S. San Martino-IST, Genova, Italy.
Fiammetta
F
Monacelli
Department of Medicine (DiMI), University of Genoa, AOU, I.R.C.C.S. San Martino-IST, Genova, Italy.
Alberto
A
Ballestrero
LeBow Institute for Myeloma Therapeutics and Jerome Lipper Center for Multiple Myeloma Research, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
Teru
T
Hideshima
LeBow Institute for Myeloma Therapeutics and Jerome Lipper Center for Multiple Myeloma Research, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
Dharminder
D
Chauhan
Department of Medicine (DiMI), University of Genoa, AOU, I.R.C.C.S. San Martino-IST, Genova, Italy.
Marco
M
Gobbi
Department of Medicine (DiMI), University of Genoa, AOU, I.R.C.C.S. San Martino-IST, Genova, Italy.
Roberto M
RM
Lemoli
LeBow Institute for Myeloma Therapeutics and Jerome Lipper Center for Multiple Myeloma Research, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
Nikhil
N
Munshi
Bing Center for Waldenstrom's Macroglobulinemia, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
Steven P
SP
Treon
LeBow Institute for Myeloma Therapeutics and Jerome Lipper Center for Multiple Myeloma Research, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
Kenneth C
KC
Anderson
D000178:Acrylamides / Q000494:pharmacology; D000225:Adenine / Q000031:analogs & derivatives; D000077329:Agammaglobulinaemia Tyrosine Kinase; D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival; D016207:Cytokines / Q000378:metabolism*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D009154:Mutation / Q000187:drug effects / Q000235:genetics*; D064549:Myeloid Cell Leukemia Sequence 1 Protein / Q000378:metabolism; D053594:Myeloid Differentiation Factor 88 / Q000235:genetics*; D016328:NF-kappa B / Q000378:metabolism; D054409:Nicotinamide Phosphoribosyltransferase / Q000378:metabolism*; D010880:Piperidines / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011720:Pyrazoles / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D019718:Receptors, CXCR4 / Q000235:genetics*; D015398:Signal Transduction / Q000187:drug effects; D008258:Waldenstrom Macroglobulinemia / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism
D000178:Acrylamides; C527276:CXCR4 protein, human; D016207:Cytokines; C507779:MYD88 protein, human; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D053594:Myeloid Differentiation Factor 88; C480543:N-(4-(1-benzoylpiperidin-4-yl)butyl)-3-(pyridin-3-yl)acrylamide; D016328:NF-kappa B; D010880:Piperidines; D011720:Pyrazoles; D011743:Pyrimidines; D019718:Receptors, CXCR4; C551803:ibrutinib; D054409:Nicotinamide Phosphoribosyltransferase; C085455:nicotinamide phosphoribosyltransferase, human; D011065:Poly(ADP-ribose) Polymerases; D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase; C000625949:BTK protein, human; D053148:Caspase 3; D000225:Adenine