Chronic inflammation is increasingly recognized as a key contributor to tumorigenesis, promoting genomic instability, immune evasion, and malignant transformation. Inflammatory cytokines are particularly involved in the pathogenesis and progression of myelodysplastic syndromes (MDS). This study examined the impact of three cytokine polymorphisms—IFNG + 874 A/T, TNFA − 857 C/T, and IL1B -31 C/T—on MDS susceptibility and clinical characteristics. A total of 105 patients with MDS and 117 healthy Japanese controls were analyzed. High-expression TNFA − 857 C/T was significantly associated with adverse risk categories by the revised International Prognostic Scoring System (high or very high: non-CC vs. CC = 36.4

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Maternal immunoglobulin (Ig)G is present in breast milk and has been shown to contribute to the development of the immune system in infants. In contrast, maternal IgG has no known effect on early childhood brain development. We found maternal IgG immunoreactivity in microglia, which are resident macrophages of the central nervous system of the pup brain, peaking at postnatal one week. Strong IgG immunoreactivity was observed in microglia in the corpus callosum and cerebellar white matter. IgG stimulation of primary cultured microglia activated the type I interferon feedback loop by Syk. Analysis of neonatal Fc receptor knockout (FcRn KO) mice that could not take up IgG from their mothers revealed abnormalities in the proliferation and/or survival of microglia, oligodendrocytes, and some types of interneurons. Moreover, FcRn KO mice also exhibited abnormalities in social behavior and lower locomotor activity in their home cages. Thus, changes in the mother-derived IgG levels affect brain development in offsprings.

Background: Integrins are cell surface proteins and major cell adhesion transmembrane receptors that play an important role in cell adhesion andmultifaceted roles in cell growth, survival, and cytoskeleton formation by transmitting stimuli from ligands to the cell. Integrin α6 acts as a receptor for laminin, an extracellular matrix, and is involved in growth, invasion, and resistance to therapy in many cancers. However, the role of integrin α6 and interaction with laminin in multiple myeloma (MM) have not yet been investigated. Therefore, we focused on the regulation of integrin α6 expression in MM cells. Additionally, we focused on the binding of integrin α6 to laminin and examined proliferation and signal transduction by laminin binding. Methods: ITGA6 expression was measured by RT-qPCR in CD138-positive plasma cells from 106 patients with MM, 13 patients with SMM, 72 patients with MGUS, 17 controls, and human myeloma cell lines (HMCL): KMS27, KMS11, KMS28BM, MM1S, KMS26, RPMI8226, and KMM1. The expression of integrin α6 on the HMCL cell surface was measured by flow cytometry. In HMCL, cell proliferation with laminin and molecules involved in signal transduction was evaluated using the CCK-8 assay and Western blotting. Results: ITGA6 mRNA expression was significantly higher in the MM group than in the control and MGUS groups (p <0.001). The median ITGA6 mRNA expression levels in the control, MGUS, SMM, and MM groups were 0.359, 2.033, 35.230, and 7.961, respectively. There were no differences in ITGA6 expression in MM samples according to ISS (p=0.772) and cytogenetic risk (p=0.371). The overall survival and progression free survival did not differ between high and low ITGA6 expressions (OS; 5.1 years vs not reached; p = 0.394, PFS; 2.1 vs 2.2 years; p = 0.523). To examine the regulation of integrin α6 expression by Myc, JQ1 and Myc inhibitors were added to the cells, and mean fluorescence intensity (MFI) decreased from 7.03 to 1.97, 6.15 to 2.54, and 5.71 to 2.31 in KMS27, KMS11, and KMS28BM, respectively, as JQ1 was added. MFI decreased from 4.23 to 3.08, 3.32 to 2.14, and 3.17 to 2.14 in KMS27, MM1S, and KMS28BM, respectively, as the Myc inhibitor was added. Since it was reported that integrin α6 expression is regulated by HIF-1α, a hypoxia-inducible factor, we examined whether integrin α6 expression is regulated by HIF-1α in HMCL. Comparing cells cultured under normal oxygen and 1% O2 conditions, the MFI of integrin α6 changed from 5.55 to 3.43, 1.43 to 1.51, 1.01 to 0.99, and 5.53 to 4.40 in KMS11, KMS26, RPMI8226, and KMM1, respectively. We examined changes in cell proliferation caused by the binding of integrin α6 to laminin. We used laminins 111, 211, 411, and 511, which are known ligands for integrin α6. Laminin 411 promoted the proliferation of KMS27 and KMS11 cells expressing integrin α6 compared to those without laminin (p=0.0207, p=0.0532). In HMCL with high integrin α6 expression, stimulation with laminin 411 increased the phosphorylation of Akt and Erk1/2. KMS27 increased phosphorylated Akt by 12-fold, phosphorylated Erk1/2 by 2-fold, and 6-fold; KMS11 increased phosphorylated Akt by 15-fold, phosphorylated Erk1/2 by 34-fold, and 11-fold. Conversely, in RPMI8226 with no integrin α6 expression, stimulation of laminin 411 resulted in a 0.6- and 1-fold increase in phosphorylated Akt and Erk1/2, respectively. Conclusion: RT-qPCR results suggest that integrin α6 may be involved in disease onset and progression. Cell surface expression of integrin α6 is regulated by Myc but not by HIF-1α. Stimulation of MM cells by laminin stimulated proliferation and caused phosphorylated Akt and Erk, suggesting that laminin is involved in MM proliferation via integrin α6. This study is a step toward developing integrin α6-targeted therapy.

INTRODUCTION:The link between DNA repair gene polymorphisms and cancer susceptibility has gained significant attention. Thus, we investigated the impact of base excision repair (BER) gene polymorphisms on acute myeloid leukemia (AML) risk and pathogenesis. METHODS:In total, 106 patients with AML and 191 healthy controls were included in the study, wherein polymorphisms in four BER genes (APEX1, MUTYH, OGG1, and XRCC1) were examined. RESULTS:Notably, the APEX1-656 T>G polymorphism exhibited a significant association with AML risk in the recessive (TT vs. TG + GG) (p = 0.046) and co-dominant models (TT vs. GG) (p = 0.02). Assessing APEX1 expression levels, APEX1 expression was elevated in the bone marrow of patients with AML compared with that in controls (p = 0.02). Subsequently, we compared the percentages of CD34+ cells between the APEX1 high or low expression groups, revealing a significant difference (high vs. low = 29.9% vs. 11.5%, p = 0.01). Additionally, we observed reduced APEX1 expression in HL60 cells differentiated with all-trans retinoic acid (p < 0.001). We hypothesized that APEX1 expression could correlate with stemness and analyzed its expression in stem and differentiated cells. CONCLUSIONS:In the GSE48558 dataset, AML cells and normal CD34+ cells expressed APEX1 at higher levels than did granulocytes (p < 0.01). Functional experiments revealed that APEX1 knockdown led to a reduction in AML cell proliferation. These findings indicated that APEX1 polymorphisms were a potential risk factor for AML and highlighted the important role of APEX1 in regulating AML cell differentiation and proliferation.

Introduction: TP53 is a tumor suppressor gene located on chromosome 17p 13.1 that plays a critical role in preventing and reducing the aggressiveness of many tumor types, including multiple myeloma (MM). 17p deletions and TP53 mutations are associated with poor clinical outcomes in patients with MM. Rho GTPase signaling is involved in cancer progression, dissemination, and chemoresistance. Among the three major Rho-GTPases (RhoA, Rac1, and Cdc42) we selected Rac1 because a previous study on lymphoma demonstrated an interaction between RaC1 and p53. In this study, we evaluated the role of Rac1 in MM, to inform novel therapy development. Materials and Methods: Bone marrow plasma cells obtained with informed consent from 114 MM patients, 70 MGUS patients, and 15 controls, and purified via anti-CD138 antibody and magnetic beads were included in this study. The study was approved by Gunma University's IRB, and followed the Declaration of Helsinki guidelines. Three human myeloma cell lines (HMCLs), KMS11, KMS26, and MM.1S, whose respective TP53 statuses were deficient, mutated, and wild type, were used. KMS26 and KMS11 cells expressing DOX-inducible wild type (WT) p53 (KMS26/Tet-on p53 and KMS11/Tet-on p53, respectively) and p53 MM.1S knockdown by shp53 were used. RAC1 mRNA levels were assessed by RT-qPCR, with ACTB serving as an endogenous control, and MM.1S as a reference sample. Protein expression levels of p53, p21, Mdm2, and Rac1 were determined via western blotting. To quantify cell proliferation or induce cell death, EdU and Annexin V assays were performed. RNA-seq was performed using an Illumina Next Seq 500. Results: KMS11/Tet-on p53 cell proliferation was not affected by p53 induction, while that of KMS26/Tet-on p53 cells was significantly reduced. Gene Ontology (GO) analysis using RNA-seq data demonstrated enhanced Rho-GTPase signaling in KMS11 compared to KMS26. RAC1 mRNA levels in purified BM plasma cells were significantly higher in patients with NDMM than in controls (p < 0.01). Rac1 inhibitor 1A-116 (50 μM) significantly reduced survival rates of both KMS11 and KMS26 cells at 72 h. This effect was more prominent in KMS11 cells than in KMS26 cells. 1A-116 also significantly reduced MM.1S cell survival at 72 h with p53 induction by MDM2 inhibitor Nutlin-3 (1 µM), or p53 knockdown by Shp53, although knockdown of WT p53 alone or Nutlin-3 alone did not affect survival. In KMS11/Tet-on p53, KMS26/Tet-on p53, and MM.1S cells, cotreatment with Nutlin-3 and 1A-116 did not increase p53, p21, or Mdm2 protein expression. In MM.1S cells with p53 knockdown, treatment with 1A-116 did not increase p53 or Mdm2 protein expression, but did increase that of p21. 1A-116 treatment significantly reduced EdU incorporation in all three HMCLs, indicating that Rac1 inhibition arrests the cell cycle, but the magnitude of this effect was attenuated in KMS11 cells. Apoptotic and dead cells, defined by annexin V-positivity and 7-AAD incorporation, were significantly increased after 1A-116 treatment in KMS11 and MM.1S cells. KMS11 and KMS26 cell survival at 72 h after treatment declined when CRBN modulators lenalidomide, pomalidomide, and iberdomide were combined with the Rac1 inhibitor 1A-116 (25µM) compared with CRBN modulator treatment alone. In contrast, Rac1 inhibitor showed no additive effect on cell survival after 24 h of bortezomib treatment in HMCLs. Median overall survival (OS) time of patients with high RAC1 mRNA expression (above median value) was significantly shortened (4.3 years vs not reached; p = 0.01), although progression free survival (PFS) was not significantly different (2.0 years vs 3.1 years; p = 0.21). In multivariate analysis, ASCT and RAC1 mRNA expression were independent prognostic factors for OS (hazard ratio [HR], 0.409; p = 0.04; RAC1 mRNA high: HR, 2.211; p = 0.02). Among patients who underwent ASCT, both OS and PFS were significantly lower in those with high RAC1 mRNA expression than in those with low RAC1 expression (OS: 5.1 years vs not reached, p = 0.02; PFS: 2.7 years vs not reached, p = 0.01). Conclusions: Rac1 affects HMCL survival regardless of p53 status and Rac1 associates with CRBN modulator sensitivity. High RAC1 mRNA expression in intramedullary plasma cells of patients with NDMM is associated with worse prognosis. Our research provides new insights for development of novel therapies targeting the Rac1 pathway to improve MM patient prognosis, including patients with p53 dysfunction.

T-cell senescence is thought to result from the age-related loss of the ability to mount effective responses to pathogens and tumor cells. In addition to aging, T-cell senescence is caused by repeated antigenic stimulation and chronic inflammation. Moreover, we demonstrated that T-cell senescence was induced by treatment with DNA-damaging chemotherapeutic agents. The characteristics of therapy-induced senescent T (TIS-T) cells and general senescent T cells are largely similar. Senescent T cells demonstrate an increase in the senescence-associated beta-galactosidase-positive population, cell cycle arrest, secretion of senescence-associated secretory phenotypic factors, and metabolic reprogramming. Furthermore, senescent T cells downregulate the expression of the co-stimulatory molecules CD27 and CD28 and upregulate natural killer cell-related molecules. Moreover, TIS-T cells showed increased PD-1 expression. However, the loss of proliferative capacity and decreased expression of co-stimulatory molecules associated with T-cell senescence cause a decrease in T-cell immunocompetence. In this review, we discuss the characteristics of senescent T-cells, including therapy-induced senescent T cells.

MicroRNAs (miRNAs and miRs) are small (19–25 base pairs) non-coding RNAs with the ability to modulate gene expression. Previously, we showed that the miR-34 family is downregulated in multiple myeloma (MM) as the cancer progressed. In this study, we aimed to clarify the mechanism of miRNA dysregulation in MM. We focused particularly on the interaction between MYC and the TP53-miR34 axis because there is a discrepancy between increased TP53 and decreased miR-34 expressions in MM. Using the nutlin-3 or Tet-on systems, we caused wild-type (WT) p53 protein accumulation in human MM cell lines (HMCLs) and observed upregulated miR-34 expression. Next, we found that treatment with an Myc inhibitor alone did not affect miR-34 expression levels, but when it was coupled with p53 accumulation, miR-34 expression increased. In contrast, forced MYC activation by the MYC-ER system reduced nutlin-3-induced miR-34 expression. We also observed that TP53 and MYC were negatively correlated with mature miR-34 expressions in the plasma cells of patients with MM. Our results suggest that MYC participates in the suppression of p53-dependent miRNA expressions. Because miRNA expression suppresses tumors, its inhibition leads to MM development and malignant transformation.

INTRODUCTION:Waldenström macroglobulinemia (WM) represents a subset of lymphoplasmacytic lymphoma (LPL) with the immunoglobulin (Ig)M paraprotein. MYD88 L265P and CXCR4 mutations are common mutations in WM patients, and mutations in ARID1A and KMT2D (MLL2) have also been reported. However, little information has been accumulated on genetic changes in LPL with other paraproteins like IgG.METHODS:We therefore aimed to evaluate genetic differences between WM and LPL with non-IgM paraprotein (non-IgM-type LPL) using targeted next-generation sequencing (NGS) in 20 Japanese patients (10 with WM, 10 with non-IgM-type LPL).RESULTS:Mutations were detected in ARID1A (10%), CXCR4 (20%), MYD88 (90%), and KMT2D (0%) for WM patients and in ARID1A (10%), CXCR4 (20%), MYD88 (70%), and KMT2D (10%) for non-IgM-type LPL patients. No significant differences were identified. No mutations were detected in NOTCH2, PRDM1, CD274 (PD-L1), PDCD1LG2 (PD-L2), RAG2, MYBBP1A, TP53, or CD79B.DISCUSSION:Mutant allele frequency in MYD88 L265P did not differ significantly between WM and non-IgM-type LPL. Most mutations detected by NGS were subclonal following MYD88 L265P, although one non-IgM-type LPL patient harbored only CXCR4 S338X mutation. Our NGS analyses reveal genetic characteristics in LPL patients and suggest genetic similarities between these two subsets of LPL, WM and non-IgM-type.

Tissue inhibitors of metalloproteinases (TIMPs) are endogenous matrix metalloproteinase inhibitors. TIMP1 is produced by cancer cells and has pleiotropic activities. However, its role and source in multiple myeloma (MM) are unclear. Here, we evaluated TIMP1 protein and mRNA levels in bone marrow (BM) plasma cells and assessed the effects of TIMP1 expression on fibroblast invasive capacity using three-dimensional spheroid cell invasion assays. TIMP1 mRNA and protein levels were elevated when patients progressed from monoclonal gammopathy of undetermined significance or smouldering myeloma to MM. Furthermore, TIMP1 levels decreased at complete response and TIMP1 protein levels increased with higher international staging. TIMP1 mRNA levels were markedly higher in extramedullary plasmacytoma and MM with t(4;14). Overall survival and post-progression survival were significantly lower in MM patients with high TIMP1 protein. Recombinant TIMP1 did not directly affect MM cells but enhanced the invasive capacity of fibroblasts; this effect was suppressed by treatment with anti-TIMP1 antibodies. Fibroblasts supported myeloma cell invasion and expansion in extracellular matrix. Overall, these results suggested that MM-derived TIMP1 induces the invasive phenotype in fibroblasts and is involved in disease progression. Further studies are required to elucidate the specific roles of TIMP1 in MM and facilitate the development of novel therapies targeting the TIMP1 pathway.

Cellular senescence is linked to a wide range of age-related diseases and can be triggered by a variety of stresses, including DNA damage. A variety of genotoxic stressors, such as anti-cancer drugs, cause DNA double-strand breaks (DSBs), which trigger the accumulation of the tumour suppressor protein p53 in the nucleus. Cellular stresses stabilize and activate the p53 signalling pathway, which regulates various cellular processes, such as apoptosis, DNA repair, and senescence. Although p53 signalling is a well-known tumour suppressor pathway, it remains unclear how it is regulated during cellular senescence. Here, we show that p53-binding protein 1 (53BP1) accumulation in the nuclear foci is required for DNA damage-induced cellular senescence via p53 activation. In human immortalized fibroblast, shRNA-mediated 53BP1 depletion decreased not only the expression of p53-target genes but also the cellular senescence induced by adriamycin treatment. Furthermore, we confirmed that DSBs trigger the hyperaccumulation of 53BP1 in the nuclear foci, which plays a key role in the regulation of cellular senescence. To prevent the accumulation of 53BP1 in the nuclear foci, we used phase separation inhibitors, and siRNA against RNF168, which accumulates at DSB loci and forms complexes with 53BP1. This blocks the formation of 53BP1 nuclear foci and DNA damage-induced cellular senescence by activating the p53 signaling pathway. In conclusion, we demonstrated that increased accumulation of 53BP1 in the nuclear foci following DNA damage activates p53 and governs cellular senescence via a liquid-liquid phase separation mechanism.

Cellular senescence is a stable cell cycle arrest, usually in response to internal and/or external stress, including telomere dysfunction, abnormal cellular growth, and DNA damage. Several chemotherapeutic drugs, such as melphalan (MEL) and doxorubicin (DXR), induce cellular senescence in cancer cells. However, it is not clear whether these drugs induce senescence in immune cells. We evaluated the induction of cellular senescence in T cells were derived from human peripheral blood mononuclear cells (PBMNCs) in healthy donors using sub-lethal doses of chemotherapeutic agents. The PBMNCs were kept overnight in RPMI 1640 medium with 2

Human endogenous retroviruses (HERVs) are retrotransposons that infect human germline cells and occupy 5-8% of the human genome. Their expression, though inhibited by mutation, deletion, and epigenetic mechanisms under normal conditions, is associated with diseases including cancer. This study aimed to clarify the association between HERVs and multiple myeloma (MM) progression. We found that HERV-K envelope (env) and long-term repeat (LTR) expression was statistically significantly higher within plasma cells in MM than in monoclonal gammopathy of undetermined significance or controls. HERV-K env knockdown increased proliferation in the MM.1S cell line and decreased the expression of the tumor suppressor genes TP53 and CDKN1A. TP53 and CDKN1A were highly expressed in MM, and their expression was correlated with HERV-K expression. HERV-K knockdown reduced apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3F, 3G, and 3H expression by 10-20% in MM.1S cells. The anti-retroviral agents nevirapine and nelfinavir suppressed proliferation and increased HERV-K expression in MM cell lines. Our results suggest that HERV-K is involved in MM progression, but its role is likely to go beyond promoting cell proliferation. Clarifying the role of HERV-K in MM will lead to the discovery of novel treatment strategies and supply new insights into MM pathogenesis.

Major histocompatibility complex class II (MHC II) is important for the adaptive immune response because MHC II presents processed antigens to a cluster of differentiation 4 (CD4)-positive T-cells. Conventional doses of chemotherapeutic agents induce tumor cell death by causing DNA double-strand breaks (DSBs). However, cellular responses caused by sub-lethal doses of chemotherapeutic agents are poorly understood. In this study, using low doses of chemotherapeutic agents, we showed that DSBs enhanced the expression of MHC II on cells that originate from antigen-presenting cells (APCs). These agents induced the MHC class II transactivator (CIITA), the master regulator of MHC II, and interferon regulatory factor 1 (IRF1), a transcription factor for CIITA. Short hairpin RNA against IRF1 suppressed chemotherapeutic agent-induced CIITA expression, whereas exogenous expression of IRF1 induced CIITA. Inhibition of ataxia-telangiectasia mutated (ATM), a DSB-activated kinase, suppressed induction of IRF1, CIITA, and MHC II. Similar results were observed by inhibiting NF-κB, a downstream target of ATM. These results suggest that DSBs induce MHC II activity via the ATM-NF-κB-IRF1–CIITA pathway in cells that intrinsically present antigens. Additionally, chemotherapeutic agents induced T-cell regulatory molecules. Our findings suggest that chemotherapeutic agents enhance the antigen presentation activity of APCs for T-cell activation.

Single-nucleotide polymorphisms (SNPs) of the IDO1 and IDO2 genes have been associated with some diseases. Here, we investigated the association of IDO1 and IDO2 SNPs with the susceptibility to multiple myeloma (MM) and their relationships with MM clinical features. We obtained genomic DNA from 100 patients with MM and 149 healthy race-matched controls and determined IDO1 promoter − 1849G/T (rs3824259) and IDO2 R248W (rs10109853) genotypes by using the polymerase chain reaction–restriction fragment length polymorphism method. The patients with MM had a significantly higher frequency of the IDO2 R248W RR genotype (high-activity type) (59.0% vs. 43.6%, odds ratio = 1.86, 95% confidence interval = 1.11–3.11, P = 0.017) compared with those in healthy controls. Patients with the IDO2 R248W RR genotype (high-activity type) were significantly younger and had a significantly lower frequency of International Staging System (ISS) stage III condition than those with the RW and WW genotypes (median 63 years vs. 69 years, P = 0.025; 15 [25.4%] vs. 50 [48.8%]). In addition, the IDO2 R248W RR genotype was significantly associated with a higher level of hemoglobin at diagnosis (mean ± standard deviation, 10.7 ± 2.36 vs. 9.27 ± 2.40 g/dL; P = 0.0032). Neither polymorphism significantly affected overall survival. Our study indicates that IDO2 R248W may be associated with the susceptibility to MM and severity of anemia.

Background and aims: Recently, genome-wide analyses have revealed mutations in spliceosome machinery associated with myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML). Single-nucleotide polymorphisms (SNPs) of serine ╱ arginine-rich splicing factor 2 (SRSF2) and splicing factor 3a subunit 1 (SF3A1) were investigated in a Japanese population of patients and healthy control group. We aimed to find associations with prognosis and pathology. Methods: We obtained genomic DNA from 99 patients with MDS, 92 patients with AML, and 172 healthy controls and detected SRSF2 (rs237057) and SF3A1 (rs2074733) genotypes using polymerase chain reaction ‒ restriction fragment length polymorphism. Results: There was no statistical significance to associate these polymorphisms with susceptibility to MDS ╱ AML. However, the SF3A1 rs2074733 TC was significantly associated with higher hemoglobin level, compared to the TT gen otype (mean ± standard deviation, 10.6 ± 1.63 vs 9.09 ± 2.19 g ╱ dL; P ＝ 0.022). In addition, patients with rs2074733 TC showed a significantly lower frequency of chromosomal abnormality [ 2 (18.2 % ) vs. 46 (53.5 % ), P ＝ 0.027]. We observed no statistical significance between these polymorphisms and clinical variables for AML, or the prognosis of MDS and AML. Conclusions: Our study indicates that the SF3A1 rs2074733 TC genotype is associated with some clinical features of MDS.

Objective Myelodysplastic syndromes (MDS), caused by various genetic mutations in hematopoietic stem cells, are associated with highly variable outcomes. Poly (ADP-ribose) polymerase-1 (PARP1) plays an important role in DNA damage repair and contributes to the progression of several types of cancer. Here, we investigated the impact of PARP1 V762A polymorphism on the susceptibility to and prognosis of MDS. Methods Samples collected from 105 MDS patients and 202 race-matched healthy controls were subjected to polymerase chain reaction-restriction fragment length polymorphism for genotyping. Results The allele and genotype frequencies of PARP1 V762A did not differ between MDS patients and the control group. However, MDS patients with the PARP1 V762A non-AA genotype, which is associated with high gene activity, had shorter overall survival rates (P = .01) than those with the AA genotype. Multivariate analysis of overall survival also revealed PARP1 V762A non-AA genotype as a poor prognostic factor (P = .02). When patients were analyzed according to treatment history, the PARP1 V762A non-AA genotype was only associated with poor survival in patients who had received treatment (P = .02). Conclusion PARP1 V762A polymorphism may be an independent prognostic factor for MDS, and a predictive biomarker for MDS treatment.

Long noncoding RNAs (lncRNAs) are deregulated in human cancers and are associated with disease progression. Plasmacytoma Variant Translocation 1 (PVT1), a lncRNA, is located adjacent to the gene MYC, which has been linked to multiple myeloma (MM). PVT1 is expressed in MM and is associated with carcinogenesis. However, its role and regulation remain uncertain. We examined PVT1/MYC expression using real-time PCR in plasma cells purified from 59 monoclonal gammopathy of undetermined significance (MGUS) and 140 MM patients. The MM cell lines KMS11, KMS12PE, OPM2, and RPMI8226 were treated with JQ1, an MYC super-enhancer inhibitor, or MYC inhibitor 10058-F4. The expression levels of PVT1 and MYC were significantly higher in MM than in MGUS (p < 0.0001) and were positively correlated with disease progression (r = 0.394, p < 0.0001). JQ1 inhibited cell proliferation and decreased the expression levels of MYC and PVT1. However, 10054-F4 did not alter the expression level of PVT1. The positive correlation between MYC and PVT1 in patients, the synchronous downregulation of MYC and PVT1 by JQ1, and the lack of effect of the MYC inhibitor on PVT1 expression suggest that the expression of these two genes is co-regulated by a super-enhancer. Cooperative effects between these two genes may contribute to MM pathogenesis and progression.

Background: Acute myeloid leukemia (AML) is a hematological malignancy characterized by the autonomous growth of immature myeloid cells with impaired differentiation and maturation. Cytokines are low-molecular-weight proteins that play a basic and fundamental role in communication within the immune system. Cytokines induce various effects such as differentiation, proliferation, hematopoiesis, and inflammation of target cells. AML is also closely associated with cytokine networks in terms of proliferation, apoptosis, and differentiation of leukemic cells. Cytokines produced by Th1 involved in cell-mediated immunity are called Th1 cytokines. Th1 cytokine includes TNF-α and IL-2. Several studies have reported that TNF-α is highly expressed in leukemia cells with AML patients. Other studies have also reported that high serum level of TNF-α of AML patients is associated with poor survival outcome. However, the association between Th1 cytokine polymorphisms: TNF-α -857C/T and IL-2-330T/G and the pathogenesis of AML is unclear. Therefore, we investigated the role of these polymorphisms in AML. Materials and Methods: This study included 101 patients with AML [male/female, 56/45; age, 15-86 years; median age, 58 years; MRC classification favorable (n = 38), intermediate (n =56), and adverse (n = 7)] and 202 healthy race-matched controls. All participants provided written informed consent. This study was approved by the Institutional Review Board of Gunma University Hospital. Genotyping was performed by the polymerase chain reaction (PCR)-restriction fragment length polymorphism method. Genotype and allele frequency were compared between patient group and control group by χ2-test. Clinical features were compared using Student's t and χ2 tests. Overall survival (OS) and leukemia free survival (LFS) were calculated using the Kaplan-Meier method. Survival curves were compared using the log-rank test. Analyses were performed using the SPSS software package ver. 25 (IBM, Armonk, NY, USA). P < 0.05 was considered to represent statistical significance. Results: TNF-α -857 C/T nonCC genotype (higher producer type) increases the risk of AML (AML vs. controls = 39.6% vs. 28.2%, OR = 1.67, 95% CI = 1.01-2.75, p = 0.045). Moreover, the frequency of TNF-α -857 C/T T allele (higher producer type) was higher in AML patients compared to controls (AML vs. controls = 24.8% vs. 16.8%, OR = 1.625, 95%CI = 1.078-2.451 p = 0.02). There was no significant difference between AML patients and controls in genotype and allele frequencies of IL-2 -330 T/G. In the analysis of clinical features, the average platelet count was significantly lower in TNF-α -857 C/T TT genotype (higher producer type) (TT vs. nonTT = 2.4±1.4 vs. 4.4±5.9, p < 0.01). TT genotype (higher producer type) was also significantly higher in frequency of MRC classification adverse (TT vs. nonTT = 30.0% vs. 4.4%, p = 0.02) and history of tumor (TT vs. nonTT = 30.0% vs. 6.6%. p =0.04). Moreover, in survival time analysis, patients with TNF-α -857 C/T TT genotype (higher producer type) had significantly shortened OS compared with patients with nonTT genotype (lower producer type) (TT vs. nonTT = 17.2 months vs not reached, p < 0.01). Patients with TT genotype (high producer type) also experienced significantly shortened LFS (TT vs. nonTT = 24.0 months vs not reached, p = 0.04). Furthermore, multivariate analysis of OS revealed TNF-α -857 C/T TT genotype (higher producer type) as an independent prognostic factor (HR = 3.01, 95% CI = 1.04-8.69, p = 0.04), like age and white blood cell count. Conclusion: These results suggest that TNF-α-857 C/T T allele (higher producer type) increases the risk of AML. Furthermore, TNF-α-857 C/T TT genotype (higher producer type) affects the poor prognosis. Therefore, these data suggest the new role of TNF-α polymorphism in AML leukemogenesis. Figure Disclosures Handa: Ono: Research Funding.