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L-GP130–induced myeloma is characterized by gammopathy with kidney damage, BM infiltration with lytic bone lesions, and high expres- sion of antiapoptotic proteins. ( A ) BM and spleen histology (H & E staining). Scale bars: 50 μ m. ( B ) X-ray analysis of a control femur (left) and a L-GP130 graft recipient femur (middle) with a lytic bone lesion (white arrow). Right: Histology from the lytic bone lesion (H & E). Scale bars: 5 mm (left and middle); 50 μ m (right). ( C ) Serum electrophoresis from representative GP130 and L-GP130 graft recipients, indicative of monoclonal gammopathy. ( D ) Ig levels, assessed by ELISA, from diseased L-GP130 graft recipient mice and age-matched control GP130 graft recipients. Shown is the geometric mean. * P < 0.05. ( E ) Representative H & E staining of cast nephropathy in the tubuli (white arrows) of an L-GP130 graft recipient mouse. Original magnification, ×400. ( F ) Ex vivo–cultured plasma cells from the BM of primary L-GP130 graft recipients (Pappenheim staining). Original magnification, ×63. ( G ) Immunoblot analysis for expression of the indicated proteins was performed on mesenterial tumors and spleen control cells. Pre, precancerous spleen.
Source publication
Multiple myeloma (MM) is a plasma cell neoplasm that results from clonal expansion of an Ig-secreting terminally differentiated B cell. Advanced MM is characterized by tissue damage that involves bone, kidney, and other organs and is typically associated with recurrent genetic abnormalities. IL-6 signaling via the IL-6 signal transducer GP130 has b...
Contexts in source publication
Context 1
... test this hypothesis, BM of 5-FU-treated donor mice was infected with L-GP130, WT GP130, or GFP control virus and transplanted into lethally irradiated syngeneic recipients ( Figure 2A). After stable hematopoietic reconstitution (Supple- mental Figure 3A), we analyzed GP130 and L-GP130 expression and activation of downstream signaling pathways in unselected white blood cells and found increased P-STAT3 as well as P-ERK in the primary L-GP130 graft recipients (Supplemental Figure 3B Figure 2D). Importantly, in BA/F3 cells, the major IL-6R/GP130 downstream signaling pathway, JAK/ STAT3 (8,14), was activated either by ectopic L-GP130 expres- sion or by IL-6 plus soluble IL-6R (sIL-6R) in a dose-dependent manner in the presence of WT GP130 (Supplemental Figure 2E). ...
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... L-GP130 graft recipient mice were next analyzed for the distribution and phenotype of the disease, with focus on features regularly observed in human MM. Moder- ate to intermediate myeloma involvement of BM and spleen was found in basically all mice analyzed ( Figure 3A and Supplemental Figure 5). Corresponding to the focal bone destruction found in human MM, X-ray analysis revealed lytic bone lesions in L-GP130 graft recipients ( Figure 3B). ...
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... ate to intermediate myeloma involvement of BM and spleen was found in basically all mice analyzed ( Figure 3A and Supplemental Figure 5). Corresponding to the focal bone destruction found in human MM, X-ray analysis revealed lytic bone lesions in L-GP130 graft recipients ( Figure 3B). In further accord with human MM, serum electrophoresis revealed monoclonal spikes and elevated class-switched IgG levels in L-GP130 graft recipients that were not detected in age-matched control GFP or GP130 graft recipi- ents ( Figure 3, C and D). ...
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... to the focal bone destruction found in human MM, X-ray analysis revealed lytic bone lesions in L-GP130 graft recipients ( Figure 3B). In further accord with human MM, serum electrophoresis revealed monoclonal spikes and elevated class-switched IgG levels in L-GP130 graft recipients that were not detected in age-matched control GFP or GP130 graft recipi- ents ( Figure 3, C and D). Notably, these serum changes were also associated with renal disease resembling human cast nephropa- thy ( Figure 3E). ...
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... further accord with human MM, serum electrophoresis revealed monoclonal spikes and elevated class-switched IgG levels in L-GP130 graft recipients that were not detected in age-matched control GFP or GP130 graft recipi- ents ( Figure 3, C and D). Notably, these serum changes were also associated with renal disease resembling human cast nephropa- thy ( Figure 3E). The morphology of tumor cells resembled human myeloma, with eccentrically located, enlarged nuclei ( Figure 3F). ...
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... these serum changes were also associated with renal disease resembling human cast nephropa- thy ( Figure 3E). The morphology of tumor cells resembled human myeloma, with eccentrically located, enlarged nuclei ( Figure 3F). Immunoblotting revealed overexpression of antiapoptotic proteins ( Figure 3G). ...
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... morphology of tumor cells resembled human myeloma, with eccentrically located, enlarged nuclei ( Figure 3F). Immunoblotting revealed overexpression of antiapoptotic proteins ( Figure 3G). In summary, the L-GP130-induced plasma cell disorder bears all key features of human myeloma with organ damage, including lytic bone lesions, monoclonal gammopathy, and kidney injury. ...
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Citations
... Epstein-Barr virus transformed lymphoblastoid cell lines from the patient showed constitutive STAT3 activation which could be inhibited by the JAK inhibitors, ruxolitinib, and tofacitinib [31]. In contrast to murine studies where B-cell-specific expression of the constitutively active artificial Lgp130 variant induced mature lymphoma and plasmacytoma [64,65], no signs of B-cell malignancies were reported for this patient. Interestingly, severe pulmonary (auto)inflammation was observed in mice with constitutive GP130 signaling in T-cells that was linked to enhanced Th17 formation [66]. ...
Genetic variants in IL6ST encoding the shared cytokine receptor for the IL-6 cytokine family GP130 have been associated with a diverse number of clinical phenotypes and disorders. We provide a molecular classification for 59 reported rare IL6ST pathogenic or likely pathogenic variants and additional polymorphisms. Based on loss- or gain-of-function, cytokine selectivity, mono- and biallelic associations, and variable cellular mosaicism, we grade six classes of IL6ST variants and explore the potential for additional variants. We classify variants according to the American College of Medical Genetics and Genomics criteria. Loss-of-function variants with (i) biallelic complete loss of GP130 function that presents with extended Stüve-Wiedemann Syndrome; (ii) autosomal recessive hyper-IgE syndrome (HIES) caused by biallelic; and (iii) autosomal dominant HIES caused by monoallelic IL6ST variants both causing selective IL-6 and IL-11 cytokine loss-of-function defects; (iv) a biallelic cytokine-specific variant that exclusively impairs IL-11 signaling, associated with craniosynostosis and tooth abnormalities; (v) somatic monoallelic mosaic constitutively active gain-of-function variants in hepatocytes that present with inflammatory hepatocellular adenoma; and (vi) mosaic constitutively active gain-of-function variants in hematopoietic and non-hematopoietic cells that are associated with an immune dysregulation syndrome. In addition to Mendelian IL6ST coding variants, there are common non-coding cis-acting variants that modify gene expression, which are associated with an increased risk of complex immune-mediated disorders and trans-acting variants that affect GP130 protein function. Our taxonomy highlights IL6ST as a gene with particularly strong functional and phenotypic diversity due to the combinatorial biology of the IL-6 cytokine family and predicts additional genotype-phenotype associations.
... In a murine retroviral transduction model, it was shown that expression of a constitutive active IL-6 signal transduction chain induced an MM-like disease that closely mimicked human MM pathology. Importantly, these tumors harbored Myc aberrations indicating that STAT3 activation and MYC collaborate to induce MM [122]. Perhaps in MM, glutamine does not only fulfil metabolic demands that are imposed and regulated by MYC, but may also act in a direct fashion by deregulating oncogenic signaling. ...
Simple Summary
Cancer is associated with metabolic changes related to increased cell proliferation and growth. These cancer-related metabolic features are largely dictated by specific oncogenes that are activated by chromosomal aberrations and epigenetic alterations in cancer cells. Multiple myeloma is an incurable plasma cell malignancy, which is characterized by recurrent chromosomal aberrations that drive the expression of established oncogenes such as MYC, Cyclin D1, FGFR3/MMSET and MAF/MAFB. In this review, we discuss the specific metabolic features of multiple myeloma plasma cells, and focus on the metabolic consequences of recurrent chromosomal aberrations, thereby providing an outline for the metabolic alterations that characterize multiple myeloma.
Abstract
Oncogene activation and malignant transformation exerts energetic, biosynthetic and redox demands on cancer cells due to increased proliferation, cell growth and tumor microenvironment adaptation. As such, altered metabolism is a hallmark of cancer, which is characterized by the reprogramming of multiple metabolic pathways. Multiple myeloma (MM) is a genetically heterogeneous disease that arises from terminally differentiated B cells. MM is characterized by reciprocal chromosomal translocations that often involve the immunoglobulin loci and a restricted set of partner loci, and complex chromosomal rearrangements that are associated with disease progression. Recurrent chromosomal aberrations in MM result in the aberrant expression of MYC, cyclin D1, FGFR3/MMSET and MAF/MAFB. In recent years, the intricate mechanisms that drive cancer cell metabolism and the many metabolic functions of the aforementioned MM-associated oncogenes have been investigated. Here, we discuss the metabolic consequences of recurrent chromosomal translocations in MM and provide a framework for the identification of metabolic changes that characterize MM cells.
... On the contrary, gp130 alone acts as a signal transducer that is responsible for a variety of biological activities [36]. gp130 signaling is an important driver of MM pathogenesis [37,38]. RPMI-8226 express both gp80 and gp130 while IL-6 is absent [39,40]. ...
Multiple myeloma is a malignant expansion of plasma cells and aggressively affects bone health. We show that P2X7 receptor altered myeloma growth, which affects primary bone cells in vitro. Expression on six human myeloma cell lines confirmed the heterogeneity associated with P2X7 receptor. Pharmacology with 2′(3′)-O-(4-benzoylbenzoyl) adenosine 5′-triphosphate (BzATP) as agonist showed dose-dependent membranal pores on RPMI-8226 (p = 0.0027) and blockade with P2X7 receptor antagonists. Ca2+ influx with increasing doses of BzATP (p = 0.0040) was also inhibited with antagonists. Chronic P2X7 receptor activation reduced RPMI-8226 viability (p = 0.0208). No apoptosis or RPMI-8226 death was observed by annexin V/propidium iodide (PI) labeling and caspase-3 cleavage, respectively. However, bromodeoxyuridine (BrdU) labelling showed an accumulation of RPMI-8226 in the S phase of cell cycle progression (61.5%, p = 0.0114) with significant decline in G0/G1 (5.2%, p = 0.0086) and G2/M (23.5%, p = 0.0015) phases. As myeloma pathology depends on a positive and proximal interaction with bone, we show that P2X7 receptor on RPMI-8226 inhibited the myeloma-induced suppression on mineralization (p = 0.0286) and reversed the excessive osteoclastic resorption. Our results demonstrate a view of how myeloma cell growth is halted by P2X7 receptor and the consequences on myeloma–osteoblast and myeloma–osteoclast interaction in vitro.
... Accordingly, and in support of the functional relevance and in vivo data, IL-6-transgenic mice on a C57BL/6 background exhibited plasmacytosis with infiltration into the spleen and LNs (19), whereas monoclonal and transplantable plasmacytomas developed only in IL-6 transgenic BALB/c mice (20). Several MM models are based on activation of the IL-6/IL-6R axis (23,42,43), underscoring its importance in proliferation and malignant transformation of late B cells. These MM mouse models are limited because of their late disease onset or low penetrance. ...
Aberrant activity of the glycoprotein 130 130/JAK/STAT3 (gp130/JAK/STAT3) signaling axis is a recurrent event in inflammation and cancer. In particular, it is associated with a wide range of hematological malignancies, including multiple myeloma and leukemia. Novel targeted therapies have only been successful for some subtypes of these malignancies, underlining the need for developing robust mouse models to better dissect the role of this pathway in specific tumorigenic processes. Here, we investigated the role of selective gp130/JAK/STAT3 activation by generating a conditional mouse model. This model targeted constitutively active, cell-autonomous gp130 activity to B cells, as well as to the entire hematopoietic system. We found that regardless of the timing of activation in B cells, constitutively active gp130 signaling resulted in the formation specifically of mature B cell lymphomas and plasma cell disorders with full penetrance, only with different latencies, where infiltrating CD138+ cells were a dominant feature in every tumor. Furthermore, constitutively active gp130 signaling in all adult hematopoietic cells also led to the development specifically of largely mature, aggressive B cell cancers, again with a high penetrance of CD138+ tumors. Importantly, gp130 activity abrogated the differentiation block induced by a B cell-targeted Myc transgene and resulted in a complete penetrance of the gp130-associated, CD138+, mature B cell lymphoma phenotype. Thus, gp130 signaling selectively provides a strong growth and differentiation advantage for mature B cells and directs lymphomagenesis specifically toward terminally differentiated B cell cancers.
... stem-like population to induce MM tumorigenesis (78). Recent adoptive transfer models have also shown promise in generating repeatable syngeneic transplantation results that promote MM (79,80). Additionally, the phenotype identified for the PCPC population needs to be translated into the human setting and phenotyped in patients specifically with relapsing/refractory myeloma, which may exhibit larger numbers of this population. ...
Multiple myeloma (MM) is an incurable plasma cell malignancy with frequent treatment failures and relapses, suggesting the existence of pathogenic myeloma stem/progenitor populations. However, the identity of MM stem cells remains elusive. We used a murine model of MM with transgenic overexpression of the unfolded protein response sensor X-box binding protein 1 (XBP1s) in the B cell compartment to define MM stem cells. We herein report that a post-germinal center, pre-plasma cell population significantly expands as MM develops. This population has the following characteristics: (a) cell surface phenotype of B220+CD19+IgM-IgD-CD138-CD80+sIgG-AA4.1+FSChi; (b) high expression levels of Pax5 and Bcl6 with intermediate levels of Blimp1 and XBP1s; (c) increased expression of aldehyde dehydrogenase, Notch1, and c-Kit; and (d) ability to efficiently reconstitute antibody-producing capacity in B cell-deficient mice in vivo. We thus have defined a plasma cell progenitor population that resembles myeloma stem cells in mice. These results provide potentially novel insights into MM stem cell biology and may contribute to the development of novel stem cell-targeted therapies for the eradication of MM.
... In addition, LGR4/Rspondin signaling facilitates signaling by Wnt ligands. Engagement of R-spondin to its receptor LGR4 induces internalization of ZNRF3/ RNF43, thereby alleviating the negative regulatory role of these E3 ligases on Wnt receptor stability silencing of β-catenin [34] and treatment with PKF115-584 [35] and CGK012 [38] showed downregulation of the Wnt target genes CCND1 (encoding cyclin D1) and MYC, which are known to play a key role in the pathogenesis of MM [4,42,43]. In addition to these established Wnt targets, several other genes involved in cell cycle regulation were also downregulated, including Aurora Kinase A (AURKA), which was identified as an important effector of Wnt-driven proliferation in MM [34]. ...
Aberrant activation of Wnt/β-catenin signaling plays a central role in the pathogenesis of a wide variety of malignancies and is typically caused by mutations in core Wnt pathway components driving constitutive, ligand-independent signaling. In multiple myelomas (MMs), however, these pathway intrinsic mutations are rare despite the fact that most tumors display aberrant Wnt pathway activity. Recent studies indicate that this activation is caused by genetic and epigenetic lesions of Wnt regulatory components, sensitizing MM cells to autocrine Wnt ligands and paracrine Wnts emanating from the bone marrow niche. These include deletion of the tumor suppressor CYLD, promotor methylation of the Wnt antagonists WIF1, DKK1, DKK3, and sFRP1, sFRP2, sFRP4, sFRP5, as well as overexpression of the co-transcriptional activator BCL9 and the R-spondin receptor LGR4. Furthermore, Wnt activity in MM is strongly promoted by interaction of both Wnts and R-spondins with syndecan-1 (CD138) on the MM cell-surface. Functionally, aberrant canonical Wnt signaling plays a dual role in the pathogenesis of MM: (I) it mediates proliferation, migration, and drug resistance of MM cells; (II) MM cells secrete Wnt antagonists that contribute to the development of osteolytic lesions by impairing osteoblast differentiation. As discussed in this review, these insights into the causes and consequences of aberrant Wnt signaling in MM will help to guide the development of targeting strategies. Importantly, since Wnt signaling in MM cells is largely ligand dependent, it can be targeted by drugs/antibodies that act upstream in the pathway, interfering with Wnt secretion, sequestering Wnts, or blocking Wnt (co)receptors.
... This model has been used to evaluate chemotherapy efficacy [69]. Constitutive activation of the IL-6 signal transducer GP130 cooperates with Myc to cause a MM-like disease in mice with lytic bone lesions, bone marrow plasma cells, and monoclonal gammopathy [70]. Anti-GP130 antibodies prevent tumor formation [71]. ...
Multiple myeloma is an invariably fatal cancer of plasma cells. Despite tremendous advances in treatment, this malignancy remains incurable in most individuals. We postulate that strategies aimed at prevention have the potential to be more effective in preventing myeloma-related death than additional pharmaceutical strategies aimed at treating advanced disease. Here, we present a rationale for the development of prevention therapy and highlight potential target areas of study.
... murine transduction-transplantation model of myeloma driven by a constitutively active allele of Gp130 encoding the common co-receptor subunit for cytokine receptors, including IL-6R also showed enhanced tumorigenesis in cooperation with c-MYC. In plasmacytomas arising from Gp130 activation alone, activating genetic alterations in the c-MYC locus were recovered in three of ten independently arising tumors (33). Combining JAK and BET inhibitors is a potentially promising strategy for other hematologic malignancies, including JAK2V617-dependent myeloproliferative neoplasms and acute lymphocytic leukemia (ALL) characterized by IL-7R activation (35)(36)(37)(38). ...
Introduction:
Bromodomain and extraterminal domain (BET) proteins regulate the expression of many cancer-associated genes and pathways; BET inhibitors have demonstrated activity in diverse models of hematologic and solid tumors. We report the preclinical characterization of INCB054329, a structurally distinct BET inhibitor that has been investigated in Phase 1 clinical trials.
Methods:
We used multiple myeloma models to investigate vulnerabilities created by INCB054329 treatment that could inform rational combinations.
Results:
In addition to c-MYC, INCB054329 decreased expression of oncogenes FGFR3 and NSD2/MMSET/WHSC1 that are deregulated in t(4;14) rearranged cell lines. The profound suppression of FGFR3 sensitized the t(4;14)-positive cell line OPM-2 to combined treatment with a fibroblast growth factor receptor inhibitor in vivo In addition, we show that BET inhibition across multiple myeloma cell lines resulted in suppressed interleukin (IL)-6 Janus kinase-signal transducers and activators of transcription (JAK-STAT) signaling. INCB054329 displaced binding of BRD4 to the promoter of IL-6 receptor (IL-6R) leading to reduced levels of IL-6R and diminished signaling through STAT3. Combination with JAK inhibitors (ruxolitinib or itacitinib) further reduced JAK-STAT signaling and synergized to inhibit myeloma cell growth in vitro and in vivo This combination potentiated tumor growth inhibition in vivo, even in the MM1.S model of myeloma that is not intrinsically sensitive to JAK inhibition alone.
Conclusion:
Preclinical data reveal insights into vulnerabilities created in myeloma cells by BET protein inhibition and potential strategies that can be leveraged in clinical studies to enhance the activity of INCB054329.
... More- over, when transfected into murine embryonic stem cells, these cells highly expressed the Oct4 gene and remained undifferentiated in the absence of LIF [59]. Ectopic expression of the L-gp130 cDNA in bone marrow cells resulted in multiple myeloma development with the characteristics of the human disease including monoclonal gammopathy, bone marrow infiltration with lytic bone lesions and protein deposition in the kidneys [60]. We have recently generated knock-in mice with the L-gp130 cDNA construct in the Rosa26 locus (Fig. 5b). ...
Interleukin-6 is a cytokine synthesized by many cells in the human body. IL-6 binds to a membrane bound IL-6R, which is only present on hepatocytes, some epithelial cells and some leukocytes. The complex of IL-6 and IL-6R binds to the ubiquitously expressed receptor subunit gp130, which forms a homodimer and thereby initiates intracellular signaling via the JAK/STAT and the MAPK pathways. IL-6R expressing cells can cleave the receptor protein to generate a soluble IL-6R (sIL-6R), which can still bind IL-6 and can associate with gp130 and induce signaling even on cells, which do not express IL-6R. This paradigm has been called IL-6 trans-signaling whereas signaling via the membrane bound IL-6R is referred to as classic signaling. We have generated several molecular tools to differentiate between IL-6 classic- and trans-signaling and to analyze the consequence of cellular IL-6 signaling in vivo.
... Activation of CCND1 is considered an early event in the pathogenesis of MM with MYC overexpression following at a later time point (Shou et al, 2000). In murine models, MYC activation, directly by overexpression or indirectly via IL6-JAK-STAT3 signalling or activationinduced-deaminase (AID) during somatic hypermutation, leads to plasma cell disorders resembling human MM (Chesi et al, 2008;Dechow et al, 2014). Recently, oncogenic MUC1 was identified to upregulate MYC in MM through b-catenin/ TCF4 (Tagde et al, 2016) and interferon regulator 4 (IRF4). ...
