THERAPEUTIC IMPACT OF TAK-1 INHIBITION ON TUMOR GROWTH AND BONE DESTRUCTION IN MYELOMA
(Abstract release date: 05/19/16)
EHA Library. Okabe M. 06/11/16; 135207; S451
Masahiro Okabe
Contributions
Contributions
Abstract
Abstract: S451
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 12:30 - 12:45
Location: Auditorium 2
Background
Multiple myeloma (MM) is still incurable with progressive bone loss. The development of novel therapeutic options yielding better survival outcomes with bone restoration is urgently needed. We have reported that Pim-2 is overexpressed in MM cells and their surrounding cells, namely bone marrow stromal cells (BMSCs) and osteoclasts, in bone lesions, and that treatment with Pim inhibitors markedly suppressed MM tumor growth while preventing bone destruction in MM-bearing animal models, indicating Pim-2 as an important therapeutic target in MM (Leukemia 2011, 2015). We recently found TGF-b-activated kinase-1 (TAK-1) as an upstream mediator responsible for Pim-2 up-regulation.
Aims
We therefore aimed to clarify the role of TAK-1 in tumor growth and bone destruction in MM and therapeutic impact of TAK-1 inhibition.
Methods
TAK-1 and Pim-2 expression and TAK-1 phosphorylation was analyzed by Western blotting. Osteoclastogenic activity was analyzed by TRAP staining as well as resorptive pit formation. Osteoblastogenesis was estimated with mineralized nodule formation by MC3T3-E1 preosteoblastic cells. The TAK-1 inhibitor LLZ1640-2 and TAK-1 siRNA were used to block TAK-1 activation and reduce TAK-1 expression, respectively. To create MM animal models, murine 5TGM1 MM cells were inoculated into the tibia in SCID mice.
Results
TAK-1 was constitutively over-expressed and phosphorylated in MM cells while only marginally in normal peripheral blood mononuclear cells. The TAK-1 inhibitor LLZ1640-2 dose-dependently suppressed cell growth, and induced caspase-dependent apoptosis in MM cells. Although TNF-a and IL-6 upregulated Pim-2 expression in MM cells, LLZ1640-2 abolished TNF-a-induced NF-κB, p38MAPK and ERK activation and IL-6-induced STAT3 activation in MM cells, leading to Pim-2 suppression. The TAK-1 inhibition also reduced the transcription factor Sp1 expression upregulated in MM cells. Because Sp1 induces the expression of various factors responsible for MM cell growth and survival, Sp1 reduction by the TAK-1 inhibition appears to cause efficaciously suppression of MM cell survival. LLZ1640-2 as well as TAK-1 knock-down decreased VCAM-1 expression and IL-6 production in BMSCs, and MM cell adhesion to BMSCs to impair BMSC support of MM cell growth. Interestingly, phosphorylation of TAK-1 was induced in BMSCs and MC3T3-E1 preosteoblastic cells by addition of cytokines known as inhibitors of osteoblastogenesis in MM, including IL-3, IL-7, TNF-a, TGF-b and activinA, as well as MM cell conditioned media (MMCM), suggesting TAK-1 as a common mediator to suppress osteoblastogenesis in MM. Furthermore, LLZ1640-2 abolished up-regulation of Pim-2, an inhibitory mediator of osteoblastogenesis, in BMSCs and MC3T3-E1 cells by MMCM to restore mineralized nodule formation. Moreover, the TAK-1 inhibition up-regulated in MC3T3-E1 cells phosphorylation of Smad1/5 and p38MAPK by BMP-2 while suppressing Smad2/3 phosphorylation by TGF-b, suggesting potentiation of BMP-2-mediated anabolic signaling. TAK-1 was also up-regulated in osteoclastic lineage cells along with osteoclastogenesis. Treatment with LLZ1640-2 suppressed the induction of c-fos and NFATc1 in RAW264.7 preosteoclastic cells as well as their osteoclastogenesis by RANK ligand. Finally, treatment with LLZ1640-2 potently suppressed MM growth in murine MM models.
Conclusion
TAK1 plays a pivotal role in MM tumor growth and bone destruction in MM. TAK1 may become an efficacious therapeutic target in MM to suppress tumor burden while restoring bone.
Session topic: New biological markers in MM
Keyword(s): Bone microenvironment, Myeloma, Osteoblast, Osteoclast
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 12:30 - 12:45
Location: Auditorium 2
Background
Multiple myeloma (MM) is still incurable with progressive bone loss. The development of novel therapeutic options yielding better survival outcomes with bone restoration is urgently needed. We have reported that Pim-2 is overexpressed in MM cells and their surrounding cells, namely bone marrow stromal cells (BMSCs) and osteoclasts, in bone lesions, and that treatment with Pim inhibitors markedly suppressed MM tumor growth while preventing bone destruction in MM-bearing animal models, indicating Pim-2 as an important therapeutic target in MM (Leukemia 2011, 2015). We recently found TGF-b-activated kinase-1 (TAK-1) as an upstream mediator responsible for Pim-2 up-regulation.
Aims
We therefore aimed to clarify the role of TAK-1 in tumor growth and bone destruction in MM and therapeutic impact of TAK-1 inhibition.
Methods
TAK-1 and Pim-2 expression and TAK-1 phosphorylation was analyzed by Western blotting. Osteoclastogenic activity was analyzed by TRAP staining as well as resorptive pit formation. Osteoblastogenesis was estimated with mineralized nodule formation by MC3T3-E1 preosteoblastic cells. The TAK-1 inhibitor LLZ1640-2 and TAK-1 siRNA were used to block TAK-1 activation and reduce TAK-1 expression, respectively. To create MM animal models, murine 5TGM1 MM cells were inoculated into the tibia in SCID mice.
Results
TAK-1 was constitutively over-expressed and phosphorylated in MM cells while only marginally in normal peripheral blood mononuclear cells. The TAK-1 inhibitor LLZ1640-2 dose-dependently suppressed cell growth, and induced caspase-dependent apoptosis in MM cells. Although TNF-a and IL-6 upregulated Pim-2 expression in MM cells, LLZ1640-2 abolished TNF-a-induced NF-κB, p38MAPK and ERK activation and IL-6-induced STAT3 activation in MM cells, leading to Pim-2 suppression. The TAK-1 inhibition also reduced the transcription factor Sp1 expression upregulated in MM cells. Because Sp1 induces the expression of various factors responsible for MM cell growth and survival, Sp1 reduction by the TAK-1 inhibition appears to cause efficaciously suppression of MM cell survival. LLZ1640-2 as well as TAK-1 knock-down decreased VCAM-1 expression and IL-6 production in BMSCs, and MM cell adhesion to BMSCs to impair BMSC support of MM cell growth. Interestingly, phosphorylation of TAK-1 was induced in BMSCs and MC3T3-E1 preosteoblastic cells by addition of cytokines known as inhibitors of osteoblastogenesis in MM, including IL-3, IL-7, TNF-a, TGF-b and activinA, as well as MM cell conditioned media (MMCM), suggesting TAK-1 as a common mediator to suppress osteoblastogenesis in MM. Furthermore, LLZ1640-2 abolished up-regulation of Pim-2, an inhibitory mediator of osteoblastogenesis, in BMSCs and MC3T3-E1 cells by MMCM to restore mineralized nodule formation. Moreover, the TAK-1 inhibition up-regulated in MC3T3-E1 cells phosphorylation of Smad1/5 and p38MAPK by BMP-2 while suppressing Smad2/3 phosphorylation by TGF-b, suggesting potentiation of BMP-2-mediated anabolic signaling. TAK-1 was also up-regulated in osteoclastic lineage cells along with osteoclastogenesis. Treatment with LLZ1640-2 suppressed the induction of c-fos and NFATc1 in RAW264.7 preosteoclastic cells as well as their osteoclastogenesis by RANK ligand. Finally, treatment with LLZ1640-2 potently suppressed MM growth in murine MM models.
Conclusion
TAK1 plays a pivotal role in MM tumor growth and bone destruction in MM. TAK1 may become an efficacious therapeutic target in MM to suppress tumor burden while restoring bone.
Session topic: New biological markers in MM
Keyword(s): Bone microenvironment, Myeloma, Osteoblast, Osteoclast
Abstract: S451
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 12:30 - 12:45
Location: Auditorium 2
Background
Multiple myeloma (MM) is still incurable with progressive bone loss. The development of novel therapeutic options yielding better survival outcomes with bone restoration is urgently needed. We have reported that Pim-2 is overexpressed in MM cells and their surrounding cells, namely bone marrow stromal cells (BMSCs) and osteoclasts, in bone lesions, and that treatment with Pim inhibitors markedly suppressed MM tumor growth while preventing bone destruction in MM-bearing animal models, indicating Pim-2 as an important therapeutic target in MM (Leukemia 2011, 2015). We recently found TGF-b-activated kinase-1 (TAK-1) as an upstream mediator responsible for Pim-2 up-regulation.
Aims
We therefore aimed to clarify the role of TAK-1 in tumor growth and bone destruction in MM and therapeutic impact of TAK-1 inhibition.
Methods
TAK-1 and Pim-2 expression and TAK-1 phosphorylation was analyzed by Western blotting. Osteoclastogenic activity was analyzed by TRAP staining as well as resorptive pit formation. Osteoblastogenesis was estimated with mineralized nodule formation by MC3T3-E1 preosteoblastic cells. The TAK-1 inhibitor LLZ1640-2 and TAK-1 siRNA were used to block TAK-1 activation and reduce TAK-1 expression, respectively. To create MM animal models, murine 5TGM1 MM cells were inoculated into the tibia in SCID mice.
Results
TAK-1 was constitutively over-expressed and phosphorylated in MM cells while only marginally in normal peripheral blood mononuclear cells. The TAK-1 inhibitor LLZ1640-2 dose-dependently suppressed cell growth, and induced caspase-dependent apoptosis in MM cells. Although TNF-a and IL-6 upregulated Pim-2 expression in MM cells, LLZ1640-2 abolished TNF-a-induced NF-κB, p38MAPK and ERK activation and IL-6-induced STAT3 activation in MM cells, leading to Pim-2 suppression. The TAK-1 inhibition also reduced the transcription factor Sp1 expression upregulated in MM cells. Because Sp1 induces the expression of various factors responsible for MM cell growth and survival, Sp1 reduction by the TAK-1 inhibition appears to cause efficaciously suppression of MM cell survival. LLZ1640-2 as well as TAK-1 knock-down decreased VCAM-1 expression and IL-6 production in BMSCs, and MM cell adhesion to BMSCs to impair BMSC support of MM cell growth. Interestingly, phosphorylation of TAK-1 was induced in BMSCs and MC3T3-E1 preosteoblastic cells by addition of cytokines known as inhibitors of osteoblastogenesis in MM, including IL-3, IL-7, TNF-a, TGF-b and activinA, as well as MM cell conditioned media (MMCM), suggesting TAK-1 as a common mediator to suppress osteoblastogenesis in MM. Furthermore, LLZ1640-2 abolished up-regulation of Pim-2, an inhibitory mediator of osteoblastogenesis, in BMSCs and MC3T3-E1 cells by MMCM to restore mineralized nodule formation. Moreover, the TAK-1 inhibition up-regulated in MC3T3-E1 cells phosphorylation of Smad1/5 and p38MAPK by BMP-2 while suppressing Smad2/3 phosphorylation by TGF-b, suggesting potentiation of BMP-2-mediated anabolic signaling. TAK-1 was also up-regulated in osteoclastic lineage cells along with osteoclastogenesis. Treatment with LLZ1640-2 suppressed the induction of c-fos and NFATc1 in RAW264.7 preosteoclastic cells as well as their osteoclastogenesis by RANK ligand. Finally, treatment with LLZ1640-2 potently suppressed MM growth in murine MM models.
Conclusion
TAK1 plays a pivotal role in MM tumor growth and bone destruction in MM. TAK1 may become an efficacious therapeutic target in MM to suppress tumor burden while restoring bone.
Session topic: New biological markers in MM
Keyword(s): Bone microenvironment, Myeloma, Osteoblast, Osteoclast
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 12:30 - 12:45
Location: Auditorium 2
Background
Multiple myeloma (MM) is still incurable with progressive bone loss. The development of novel therapeutic options yielding better survival outcomes with bone restoration is urgently needed. We have reported that Pim-2 is overexpressed in MM cells and their surrounding cells, namely bone marrow stromal cells (BMSCs) and osteoclasts, in bone lesions, and that treatment with Pim inhibitors markedly suppressed MM tumor growth while preventing bone destruction in MM-bearing animal models, indicating Pim-2 as an important therapeutic target in MM (Leukemia 2011, 2015). We recently found TGF-b-activated kinase-1 (TAK-1) as an upstream mediator responsible for Pim-2 up-regulation.
Aims
We therefore aimed to clarify the role of TAK-1 in tumor growth and bone destruction in MM and therapeutic impact of TAK-1 inhibition.
Methods
TAK-1 and Pim-2 expression and TAK-1 phosphorylation was analyzed by Western blotting. Osteoclastogenic activity was analyzed by TRAP staining as well as resorptive pit formation. Osteoblastogenesis was estimated with mineralized nodule formation by MC3T3-E1 preosteoblastic cells. The TAK-1 inhibitor LLZ1640-2 and TAK-1 siRNA were used to block TAK-1 activation and reduce TAK-1 expression, respectively. To create MM animal models, murine 5TGM1 MM cells were inoculated into the tibia in SCID mice.
Results
TAK-1 was constitutively over-expressed and phosphorylated in MM cells while only marginally in normal peripheral blood mononuclear cells. The TAK-1 inhibitor LLZ1640-2 dose-dependently suppressed cell growth, and induced caspase-dependent apoptosis in MM cells. Although TNF-a and IL-6 upregulated Pim-2 expression in MM cells, LLZ1640-2 abolished TNF-a-induced NF-κB, p38MAPK and ERK activation and IL-6-induced STAT3 activation in MM cells, leading to Pim-2 suppression. The TAK-1 inhibition also reduced the transcription factor Sp1 expression upregulated in MM cells. Because Sp1 induces the expression of various factors responsible for MM cell growth and survival, Sp1 reduction by the TAK-1 inhibition appears to cause efficaciously suppression of MM cell survival. LLZ1640-2 as well as TAK-1 knock-down decreased VCAM-1 expression and IL-6 production in BMSCs, and MM cell adhesion to BMSCs to impair BMSC support of MM cell growth. Interestingly, phosphorylation of TAK-1 was induced in BMSCs and MC3T3-E1 preosteoblastic cells by addition of cytokines known as inhibitors of osteoblastogenesis in MM, including IL-3, IL-7, TNF-a, TGF-b and activinA, as well as MM cell conditioned media (MMCM), suggesting TAK-1 as a common mediator to suppress osteoblastogenesis in MM. Furthermore, LLZ1640-2 abolished up-regulation of Pim-2, an inhibitory mediator of osteoblastogenesis, in BMSCs and MC3T3-E1 cells by MMCM to restore mineralized nodule formation. Moreover, the TAK-1 inhibition up-regulated in MC3T3-E1 cells phosphorylation of Smad1/5 and p38MAPK by BMP-2 while suppressing Smad2/3 phosphorylation by TGF-b, suggesting potentiation of BMP-2-mediated anabolic signaling. TAK-1 was also up-regulated in osteoclastic lineage cells along with osteoclastogenesis. Treatment with LLZ1640-2 suppressed the induction of c-fos and NFATc1 in RAW264.7 preosteoclastic cells as well as their osteoclastogenesis by RANK ligand. Finally, treatment with LLZ1640-2 potently suppressed MM growth in murine MM models.
Conclusion
TAK1 plays a pivotal role in MM tumor growth and bone destruction in MM. TAK1 may become an efficacious therapeutic target in MM to suppress tumor burden while restoring bone.
Session topic: New biological markers in MM
Keyword(s): Bone microenvironment, Myeloma, Osteoblast, Osteoclast
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