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Cancer cell–intrinsic properties caused by oncogenic mutations have been well characterized; however, how specific oncogenes and tumor suppressors impact the tumor microenvironment (TME) is not well understood. Here, we present a novel non–cell-autonomous function of the retinoblastoma (RB) tumor suppressor in controlling the TME. RB inactivation stimulated tumor growth and neoangiogenesis in a syngeneic and orthotropic murine soft-tissue sarcoma model, which was associated with recruitment of tumor-associated macrophages (TAM) and immunosuppressive cells such as Gr1+CD11b+ myeloid-derived suppressor cells (MDSC) or Foxp3+ regulatory T cells (Treg). Gene expression profiling and analysis of genetically engineered mouse models revealed that RB inactivation increased secretion of the chemoattractant CCL2. Furthermore, activation of the CCL2–CCR2 axis in the TME promoted tumor angiogenesis and recruitment of TAMs and MDSCs into the TME in several tumor types including sarcoma and breast cancer. Loss of RB increased fatty acid oxidation (FAO) by activating AMP-activated protein kinase that led to inactivation of acetyl-CoA carboxylase, which suppresses FAO. This promoted mitochondrial superoxide production and JNK activation, which enhanced CCL2 expression. These findings indicate that the CCL2–CCR2 axis could be an effective therapeutic target in RB-deficient tumors.Significance:These findings demonstrate the cell-nonautonomous role of the tumor suppressor retinoblastoma in the tumor microenvironment, linking retinoblastoma loss to immunosuppression.

RPKM value of CCL2, CCL5 and RB1 gene in each breast cancer cell line based on Cancer Cell Line Encyclopedia (CCLE) database.

This file contains additional data to further support the conclusions in the main manuscript. Supplementary figure S1 shows the data related to figure 1,2,3. Supplementary figure S2 shows the data related to figure 4,5,6.

Tennis is a popular leisure sport, and studies have indicated that playing tennis regularly provides many health benefits. We aimed to clarify the characteristics of physical activity during beginner– level group tennis lessons and daily physical activity of the participants. Physical activity was measured using an accelerometer sensor device for four weeks, including the 80–minute duration tennis lessons held twice a week. Valid data were categorized for tennis and non-tennis days. The mean physical activity intensity during the tennis lesson was 3.37 METs. The mean ratio of short-bout rest periods to the tennis lesson time in 90 and 120 seconds was 7% and 4%, respectively. Compared to the day of tennis lessons, the mean physical activity intensity and steps taken were significantly lower on days with no tennis (p p

Bendamustine and rituximab (BR) are widely used in patients with follicular lymphoma (FL) previously treated with conventional immunochemotherapy, but the role of consolidation radioimmunotherapy in these patients is unknown. This study evaluated the efficacy and safety of consolidation with 90 Yttrium-ibritumomab tiuxetan (90 Y-IT) after re-induction therapy with BR in patients with previously treated FL. This study included adult patients with relapsed FL who had undergone one or two prior therapies. Re-induction therapy with BR was administered every 4 weeks up to 4-6 cycles. If patients achieved at least partial response, 90 Y-IT was administered as consolidation therapy. The primary endpoint was 2-year progression-free survival (PFS) after consolidation. A total of 24 FL patients (median age 60 years) who had undergone one (n = 17) or two (n = 7) prior treatments received BR. After BR therapy, 22 patients proceeded to consolidation with 90 Y-IT, resulting in an overall 88% response rate to the protocol treatment. Within a median observation period of 46.8 months, the estimated 2-year PFS rate after the consolidation among the 22 patients receiving 90 Y-IT was 59% (95% confidence interval [CI], 38%-77%). Patients whose remission after previous treatment had lasted ≥2 years had a significantly higher 2-year PFS rate than patients whose remission after previous treatment had been

Wako, T., Yoshida, A., Kato, J. et al. Human metaphase chromosome consists of randomly arranged chromatin fibres with up to 30-nm diameter. Sci Rep 10, 8948 (2020). https://doi.org/10.1038/s41598-020-65842-z., During cell division, mitotic chromosomes assemble and are equally distributed into two new daughter cells. The chromosome organisation of the two chromatids is essential for even distribution of genetic materials. Although the 11-nm fibre or nucleosome structure is well-understood as a fundamental fibrous structure of chromosomes, the reports on organisation of 30-nm basic chromatin fibres have been controversial, with debates on the contribution of 30-nm or thicker fibres to the higher order inner structure of chromosomes. Here, we used focused ion beam/scanning electron microscopy (FIB/SEM) to show that both 11-nm and 30-nm fibres are present in the human metaphase chromosome, although the higher-order periodical structure could not be detected under the conditions employed. We directly dissected the chromosome every 10-nm and observed 224 cross-section SEM images. We demonstrated that the chromosome consisted of chromatin fibres of an average diameter of 16.9-nm. The majority of the chromatin fibres had diameters between 5 and 25-nm, while those with 30-nm were in the minority. The reduced packaging ratio of the chromatin fibres was detected at axial regions of each chromatid. Our results provide a strong basis for further discussions on the chromosome higher-order structure.

Bendamustine and rituximab (BR) are widely used in patients with follicular lymphoma (FL) previously treated with conventional immunochemotherapy, but the role of consolidation radioimmunotherapy in these patients is unknown. This study evaluated the efficacy and safety of consolidation with

Although allogeneic hematopoietic stem cell transplantation (allo-HSCT) is considered a potentially curative therapy for patients with multiple myeloma, the role of allo-HSCT remains unclear in the novel agent era. We conducted a retrospective study of 65 patients with multiple myeloma who underwent allo-HSCT at 19 institutions from 2009 to 2016. Patients received a median of 3 (range, 1 to 7) lines of prior therapy, including at least 1 novel agent, except for autologous HSCT. The 3-year progression-free survival (PFS) and overall survival (OS) rates were 18.8% (95% confidence interval [CI], 9.6% to 30.3%) and 47.2% (95% CI, 33.9% to 59.4%), respectively. In a multivariate analysis, an age ≥50 years and less than a very good partial response (VGPR) before allo-HSCT were independent significant adverse factors for PFS (hazard ratio [HR], 2.30, P = .0063; HR, 2.86; P = .0059) and OS (HR, 2.37, P = .013; and HR, 2.74; P = .040). In contrast, the 3-year PFS and OS rates in patients50 years of age who achieved a VGPR or better before allo-HSCT were 64.3% (95% CI, 29.8% to 85.1%) and 80.2% (95% CI, 40.3% to 94.8%), respectively. The overall response rate was 86.4% (95% CI, 75.0% to 94.0%). The proportion of VGPR or better increased from 29% before allo-HSCT to 71% after allo-HSCT. The nonrelapse mortality at 3 years was 23.4% (95% CI, 13.8% to 34.4%). Only an age ≥50 years was associated with higher nonrelapse mortality (HR, 4.71; P = .015). We showed that allo-HSCT is feasible for heavily pretreated patients with multiple myeloma, even in the novel agent era. Allo-HSCT in particular is a promising therapy for young and chemosensitive patients.

Here, we present the case of a 72-year-old male who presented with swelling, stiffness, and dysesthesia in the bilateral fingers, wrists, and ankles. Although rheumatoid arthritis was initially suspected, laboratory tests were negative for rheumatoid factor (RF) and anti-cyclic citrullinated peptide (CCP) antibody. Based on the findings of immune globulin G (IgG)-λ M proteins and 26% plasma cells in the bone marrow, multiple myeloma was diagnosed. Joint sonography revealed thickening of the tendon synovial sheaths around the bilateral wrist joints, palmar flexor tendon sheaths, and extensor digitorum tendon sheaths, and magnetic resonance imaging (MRI) revealed soft tissue masses around the bilateral hip joints. Carpal tunnel syndrome associated with amyloid arthritis was suspected. Amyloid deposits were observed in synovectomy specimens, and the patient was then diagnosed with amyloid arthritis. He had concurrent pulmonary fibrosis, and treatment with lenalidomide/dexamethasone (Ld therapy) was initiated. The symptoms in the bilateral fingers, wrists, and ankles improved with the treatment course, and joint sonography revealed that thickening of tendon sheath and soft tissue masses disappeared after seven courses of Ld therapy. However, MRI still revealed soft tissue masses around the bilateral hip joints. In patients with joint symptoms that do not fulfill the diagnostic criteria for rheumatoid arthritis, differentiation with amyloid arthritis is necessary.

Human herpesvirus-8-unrelated primary effusion lymphoma characterized by lymphomatous effusion without nodal lesions occasionally exhibits spontaneous remission. To elucidate the factors associated with a good prognosis, this study analyzed the clinical parameters of four patients treated in the department and 109 patients reported in case reports. The median age was 71 years and the median overall survival was 20 months. Patients possessing two independent favorable factors, an elderly status (≥ 70 years) and low serum lactate dehydrogenase ( 500 IU/L) showed a markedly higher 1-year survival than patients lacking either of the two factors in the absence of chemotherapy (94% vs 20%, p = 3 × 10(-5)), which was similarly observed in the chemotherapy group (94% vs 51%, p = 0.002). The use of rituximab was also a strong predictor of survival (89% vs 49%, p = 7 × 10(-6)). Elderly patients not exhibiting an increased lactate dehydrogenase may represent a benign sub-group of effusion lymphoma, which do not require chemotherapy to achieve remission.

Mutations in RB and PTEN are linked to castration resistance and poor prognosis in prostate cancer. Identification of genes that are regulated by these tumor suppressors in a context that recapitulates cancer progression may be beneficial for discovering novel therapeutic targets. Although various genetically engineered mice thus far provided tumor models with various pathological stages, they are not ideal for detecting dynamic changes in gene transcription. Additionally, it is difficult to achieve an effect specific to tumor progression via gain of functions of these genes. In this study, we developed an in vitro model to help identify RB- and PTEN-loss signatures during the malignant progression of prostate cancers. Trp53-/- ; Rbf/f , Trp53-/- ; Ptenf/f , and Trp53-/- ; Rbf/f ; Ptenf/f prostate epithelial cells were infected with AD-LacZ or AD-Cre. We found that deletion of Rb, Pten or both stimulated prostasphere formation and tumor development in immune-compromised mice. The GO analysis of genes affected by the deletion of Rb or Pten in Trp53-/- prostate epithelial cells identified a number of genes encoding cytokines, chemokines and extracellular matrix remodeling factors, but only few genes related to cell cycle progression. Two genes (Il-6 and Lox) were further analyzed. Blockade of Il-6 signaling and depletion of Lox significantly attenuated prostasphere formation in 3D culture, and in the case of IL-6, strongly suppressed tumor growth in vivo. These findings suggest that our in vitro model may be instrumental in identifying novel therapeutic targets of prostate cancer progression, and further underscore IL-6 and LOX as promising therapeutic targets. © 2015 Wiley Periodicals, Inc.

Cancer cell–intrinsic properties caused by oncogenic mutations have been well characterized; however, how specific oncogenes and tumor suppressors impact the tumor microenvironment (TME) is not well understood. Here, we present a novel non–cell-autonomous function of the retinoblastoma (RB) tumor suppressor in controlling the TME. RB inactivation stimulated tumor growth and neoangiogenesis in a syngeneic and orthotropic murine soft-tissue sarcoma model, which was associated with recruitment of tumor-associated macrophages (TAM) and immunosuppressive cells such as Gr1+CD11b+ myeloid-derived suppressor cells (MDSC) or Foxp3+ regulatory T cells (Treg). Gene expression profiling and analysis of genetically engineered mouse models revealed that RB inactivation increased secretion of the chemoattractant CCL2. Furthermore, activation of the CCL2–CCR2 axis in the TME promoted tumor angiogenesis and recruitment of TAMs and MDSCs into the TME in several tumor types including sarcoma and breast cancer. Loss of RB increased fatty acid oxidation (FAO) by activating AMP-activated protein kinase that led to inactivation of acetyl-CoA carboxylase, which suppresses FAO. This promoted mitochondrial superoxide production and JNK activation, which enhanced CCL2 expression. These findings indicate that the CCL2–CCR2 axis could be an effective therapeutic target in RB-deficient tumors. Significance: These findings demonstrate the cell-nonautonomous role of the tumor suppressor retinoblastoma in the tumor microenvironment, linking retinoblastoma loss to immunosuppression.

We established an in vitro cell culture system to determine novel activities of the retinoblastoma (Rb) protein during tumor progression. Rb depletion in p53-null mouse-derived soft tissue sarcoma cells induced a spherogenic phenotype. Cells retrieved from Rb-depleted spheres exhibited slower proliferation and less efficient BrdU incorporation, however, much higher spherogenic activity and aggressive behavior. We discovered six miRNAs, including mmu-miR-18a, -25, -29b, -140, -337, and -1839, whose expression levels correlated tightly with the Rb status and spherogenic activity. Among these, mmu-miR-140 appeared to be positively controlled by Rb and to antagonize the effect of Rb depletion on spherogenesis and tumorigenesis. Furthermore, among genes potentially targeted by mmu-miR-140, Il-6 was upregulated by Rb depletion and downregulated by mmu-mir-140 overexpression. Altogether, we demonstrate the possibility that mmu-mir-140 mediates the Rb function to downregulate Il-6 by targeting its 3'-untranslated region. Finally, we detected the same relationship among RB, hsa-miR-140 and IL-6 in a human breast cancer cell line MCF-7. Because IL-6 is a critical modulator of malignant features of cancer cells and the RB pathway is impaired in the majority of cancers, hsa-miR-140 might be a promising therapeutic tool that disrupts linkage between tumor suppressor inactivation and pro-inflammatory cytokine response., Supplementary Table1 and Supplementary Table2: We offer the table data with an Excel file

Retinoblastoma (RB) protein inactivation during tumor progression is often associated with acquisition of immature phenotypes and resistance to therapy. Determination of an RB inactivation signature in a context of gaining undifferentiated phenotype in a p53-null sarcoma system revealed a critical role for interleukin (IL)-6. Using a Gene Set Enrichment Analysis (GSEA), we discovered that poorly differentiated breast cancers are enriched for this RB inactivation signature. Accelerated IL-6 secretion following RB inactivation in an RB-intact luminal-type breast cancer cell line MCF-7 promoted a positive feed forward loop between IL-6 and STAT3 driving tumor growth and endocrine therapy resistance. In addition, some of RB-intact basal-like type breast cancer cell lines exhibited a similar phenotype following RB depletion. The mechanism whereby RB inactivation increases IL-6 production in MCF-7 cells appeared to involve fatty acid oxidation (FAO)-dependent mitochondrial metabolism and c-Jun NH(2)-terminal kinase (JNK). In addition, IL-6, via STAT3-mediated feedback to mitochondria, autonomously adjusts mitochondrial superoxide to levels suitable to maintain stem cell-like activity. The gene expression profile of luminal-type breast cancer patients with low RB expression revealed high enrichment of genes involved in mitochondrial respiration and downstream targets of IL-6. These findings unveiled an unexpected strategy whereby RB suppresses malignant features of cancer cells through metabolic reprogramming and cell-autonomous inflammation.

Mutations in RB and PTEN are linked to castration resistance and poor prognosis in prostate cancer. Identification of genes that are regulated by these tumor suppressors in a context that recapitulates cancer progression may be beneficial for discovering novel therapeutic targets. Although various genetically engineered mice thus far provided tumor models with various pathological stages, they are not ideal for detecting dynamic changes in gene transcription. Additionally, it is difficult to achieve an effect specific to tumor progression via gain of functions of these genes. In this study, we developed an in vitro model to help identify RB- and PTEN-loss signatures during the malignant progression of prostate cancers. Trp53

We have applied AFM-IR to characterize the chemical structure of polyimide film near the interface with a copper substrate as a model system of an industrial device. AFM-IR requires sample preparation of slicing a sample to be a thin film before its measurement. We have adopted FIB processing as the sample preparation method because a microtome is difficult to slice the cupper substrate. From the comparison between the spectra obtained by traditional FT-IR-ATR, AFM-IR with microtome preparation and FIB processing, we conclude that the damage on polyimide due to ion beam irradiation is so negligible as to characterize the chemical structure of polyimide. We have found the production of carboxylate near the interface by AFM-IR spectral measurement. Furthermore, we have succeeded in visualizing the distribution of carboxylate with a spatial resolution of a few hundreds nanometers which has been difficult to be achieved by a traditional microscopic IR spectroscopy. AFM-IR in conjunction with FIB processing is a useful and promising tool for nanometer scale characterization of chemical structure of polymer materials in organic/inorganic composites such as micro-structured semiconductor devices.

The vapor-phase aldol condensation of propionic acid (PA) with formaldehyde (HCHO) to form methacrylic acid (MAA) was performed with a PA/HCHO molar ratio of 2 using various silica-supported metal phosphate catalysts. In the absence of water, a silica-supported tin phosphate with Sn/Si/P atomic ratio of 1/8/2.3 showed the same level of performance as a silica-supported vanadium phosphate with V/Si/P atomic ratio of 1/8/2.2 did. The one-pass yield of MAA reached 58% based on the HCHO feed. In the presence of water, the performances of almost all catalysts fell. However, the Sn/Si/P catalyst showed a relatively high performance even in the presence of water. The performance is almost identical to that obtained from a silica-supported cesium oxide catalyst.

Acute myeloid leukaemia (AML) is a heterogeneous neoplastic disorder in which a subset of cells function as leukaemia-initiating cells (LICs). In this study, we prospectively evaluated the leukaemia-initiating capacity of AML cells fractionated according to the expression of a nucleolar GTP binding protein, nucleostemin (NS). To monitor NS expression in living AML cells, we generated a mouse AML model in which green fluorescent protein (GFP) is expressed under the control of a region of the NS promoter (NS–GFP). In AML cells, NS–GFP levels were correlated with endogenous NS mRNA. AML cells with the highest expression of NS–GFP were very immature blast-like cells, efficiently formed leukaemia colonies in vitro , and exhibited the highest leukaemia-initiating capacity in vivo . Gene expression profiling analysis revealed that cell cycle regulators and nucleotide metabolism-related genes were highly enriched in a gene set associated with leukaemia-initiating capacity that we termed the ‘leukaemia stem cell gene signature’. This gene signature stratified human AML patients into distinct clusters that reflected prognosis, demonstrating that the mouse leukaemia stem cell gene signature is significantly associated with the malignant properties of human AML. Further analyses of gene regulation in leukaemia stem cells could provide novel insights into diagnostic and therapeutic approaches to AML.

Background: Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare malignancy of plasmacytoid dendritic cells as classified by the World Health Organization (WHO), and is characterized by distinct clinical, pathological and genetic features. BPDCN used to be diagnosed as blastic NK-cell lymphoma or CD4+/CD56+ hematodermic neoplasm. Its prognosis is extremely poor with a median overall survival (OS) of about 1 year. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) in remission has shown encouraging results for patients with BPDCN. However, the role of high-dose chemotherapy followed by autologous stem cell transplantation (HDT/ASCT) has not been reported. Methods: We surveyed the data of the Transplant Registry Unified Management Program of the Japan Society for Hematopoietic Cell Transplantation and identified 109 patients with an original diagnosis of BPDCN, blastic NK-cell or lymphoblastic lymphoma of NK-cell lineage. These diagnoses had been made by a pathologist at each institution in accordance with the WHO classification or previous lymphoma classification systems. Then, we sent out questionnaires to collect additional data on clinical presentation, prior therapies and update of follow-up, in addition to pathology and flow cytometry reports. The diagnosis of BPDCN was made with clinicopathological review by a consensus panel. Results: Of the 109 identified patients, additional data were obtained for 83. After central clinicopathological review, 58 patients were excluded from analysis due to disease other than BPDCN or the absence of important clinical data. Finally, the diagnosis of BPDCN was confirmed in 25 patients (allo-HSCT, N = 14; HDT/ASCT, N = 11). The median age at HSCT was 58 (range, 17-67) years and male patients were predominant (80%). Involvements of skin and bone marrow at diagnosis were observed in 88% and 68% of patients, respectively. The induction chemotherapy regimen was as follows: non-Hodgkin lymphoma (NHL)-like (e.g. cyclophosphamide, doxorubicin, vincristine, prednisone [CHOP]-like regimens; or ifosfamide/etoposide-based regimens) (N = 11), acute lymphoblastic leukemia (ALL)-like (N = 10) and acute myeloid leukemia (AML)-like (N = 4). The rate of initial response to induction chemotherapy was high, with a complete remission (CR) rate of 96%, except for one patient who experienced progressive disease in the central nervous system during induction therapy with an ALL-like regimen. The median time from diagnosis to HSCT was 6 months (range, 2-22). Among the patients who underwent allo-HSCT, myeloablative (MAC) and reduced-intensity (RIC) regimens were used in 8 (57%) and 6 (43%) patients, respectively. The donor of 1st allogeneic transplant was a relative in 7, unrelated bone marrow in 7 and cord blood in 1. Regarding the disease status at transplantation, all 11 patients underwent HDT/ASCT in CR1 whereas, among patients who underwent allo-HSCT, 12 were in CR1 (N=10)/CR2 (N =2) and two had disease. With a median follow-up of 53.5 months in surviving patients, OS and progression-free survival (PFS) at 4 years were 65% and 58%, respectively (Figure 1). The OS at 4 years for patients who received HDT/ASCT and allo-HSCT were 82% and 53%, respectively (P = 0.11, Figure 2). The PFS at 4 years for patients who received HDT/ASCT and allo-HSCT were 73% and 48%, respectively (P = 0.14, Figure 2). The three induction regimen groups (NHL-like, ALL-like and AML-like regimens) had similar OS rates: 62%, 70% and 67% at 4 years, respectively (P = 0.86). OS did not differ significantly between MAC and RIC groups (OS at 4 years: 45% vs. 60%, P = 0.31). There was no non-relapse mortality within 100 days after HSCT. Seven patients (28%) relapsed at a median of 10 months (range, 3-21) after HSCT (MAC allo-HSCT, N = 2; reduced-intensity conditioning allo-HSCT, N = 3; HDT/ASCT, N = 2). Notably, there was no relapse in patients without bone marrow (BM) infiltration at diagnosis in the HDT/ASCT treatment group. After allo-HSCT, grade 2-4 acute GVHD was observed in 5 patients (20%). Conclusions: As previous reports have shown, HSCT can achieve long-term survival in patients with BPDCN. In addition, the result of HDT/ASCT in CR1 seems to be promising and deserves further evaluation in the settings of prospective trials. Meanwhile, we need to clarify who could be cured by ASCT and need to improve outcome for high-risk patients, such as those with BM invasion. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

Background and aims: Resveratrol is a natural occurring polyphenol that has various health promoting properties including anti-inflammatory, anti-aging and heart protective effects, and it has also shown a promising anticancer potential (reviewed in Nature Reviews Drug Discovery, 2006). We recently reported that resveratrol may promote tumor immunosurveillance by inducing NKG2D ligands in tumor cells thereby sensitizing the transformed cells to NK cells killing (Cancer Science, 2013). Notably, despite the fact that resveratrol has been extensively studied in several in vitro and preclinical models, little is still known about its in vivo effects in humans. Based on preclinical data, we therefore initiated a phase I clinical trial to explore the safety of repeated doses of resveratrol in Japanese healthy volunteers and to also evaluate its effects on circulating immune cells. Methods: In this study, we evaluated the cellular and molecular immune response in 14 healthy volunteers at baseline and after the oral administration of both resveratrol monomer at 1 Gr/day in seven individuals and resveratrol dimer, given to 7 subjects at 500 mg/day for 28 days. We obtained blood specimens at baseline, and then every 2 weeks for a total of 12 weeks. We assessed the plasma levels of 13 cytokines and chemokines by a luminex assay. In addition, we performed high-performance liquid chromatography-UV to measure resveratrol and its metabolites in both plasma and urine samples. We also evaluated the lymphocyte phenotypes by flow cytometry. Results: The consumption of resveratrol monomer and resveratrol dimer did not cause any adverse events in the individuals evaluated in this study and the plasma levels of resveratrol and its metabolites could thus be accurately measured in both plasma and urine. Consistent with their reported antioxidant properties, the administration of resveratrol monomer (1.5 fold, p=0.01) or resveratrol dimer (2 fold p=0.01) resulted in a significant increase in the antioxidant activity of the plasma compared with the corresponding antioxidant baseline activity. The administration of resveratrol monomer, but not resveratrol dimer, resulted in an increase in the number of CD3- CD56+ and NKG2D+ circulating lymphocytes (p=0.05). Conversely, the circulating numbers of monocytes, B and T lymphocytes remained essentially unchanged throughout the course of the study. Phenotypic studies revealed a significant increase in the expression level of NKG2D receptor on NK cells (p=0.01) and gamma delta T cells (p=0.05) in the individuals receiving either resveratrol dimer or monomer. The activator receptor DNAM-1 was also upregulated in the NK cells obtained from individuals receiving resveratrol. Other NK cells receptors including NKp46, NKp30 and NKG2A remained essentially unchanged. Protein profiling demonstrated significant increases in the circulating levels of immune-related cytokines including CXCL10 ( P=0.01), and MCP-1 (P=0.001), which became apparent by the second week after resveratrol administration. In contrast, the circulating levels of IL-6, IL-10, IL-15, VEGF, and ILR1a remained at baseline levels after resveratrol administration. Conclusions: This is the first study to characterize the effects of resveratrol administration on human circulating immune cells in vivo, and these findings demonstrate that resveratrol clearly has some biological effects on immune cells. Disclosures No relevant conflicts of interest to declare.

Background: Inactivation of Retinoblastoma 1 (RB1) gene product is implicated in progression of breast, prostate and many other cancers. It has been shown that RB1 expression level is correlated with the subtypes and sensitivity to treatment in breast cancers. However, directly linking RB1 status to clinical behaviors in cancers has been hampered by heterogeneity in human cancers. To clarify the mechanism by which RB1 inactivation affects clinical courses, we obtained the gene expression profile by Rb1 depletion under genetically homogeneous conditions, and analyzed the correlation between Rb1 loss signatures and clinical behaviors in breast cancers. Methods: Trp53-null mouse derived tumor cells were transduced with shRNA of either control or Rb1, and whole RNA sequencing was performed. We compared the average between control samples and Rb1 knockdown samples. We extracted differentially expressed genes, and obtained Rb1 loss signatures by using homologous genes. Based on these signatures, we performed KEGG and GO analyses, detected overlapping genes using Mammaprit, OncotypeDX, and other reported breast cancer signatures. Also we investigated relevance to subtype, treatment sensitivity, and prognosis of breast cancer. Results: Some genes were found to be overlapped in Rb1 loss signatures and previously reported breast cancer signatures. Also the genes affected by Rb1 knockdown were included in signatures featuring high grade and poor prognosis in breast cancer. Conclusion:Rb1 depletion itself may associate clinical course in breast cancer.

Abstract 1343 Natural killer (NK)-cell malignancies, particularly aggressive NK-cell leukemias/lymphomas, have poor prognoses. Although recent regimens that include L-asparaginase substantially improve outcomes, novel therapeutic approaches are needed to improve clinical responses. Recent reports have shown that the signal transducer and activator of transcription 3 (STAT3) pathway is critical for proliferation and survival of malignant NK cells. Resveratrol is a naturally-occurring polyphenol that has been extensively studied for its anti-inflammatory, cardioprotective, and anti-cancer activities. In this study, we investigated the potential anti-tumor activities of resveratrol against the NK cell lines KHYG-1, NKL, NK-92, and NK-YS. Resveratrol significantly suppressed cell proliferation in a dose- and time-dependent manner in these four cell lines. Flow cytometry analysis with annexin V/propidium iodide staining showed a variable but consistent induction of apoptosis in the four cell lines treated with resveratrol for 48 hours, ranging from 57.1±6.9% of apoptosis in the L-asparaginase resistant cell line KHYG-1 to 53.4±9.7%, 28.8±3.3%, and 51.7±6.7% in NKL, NK-92, and NK-YS cells respectively (Fig. 1a). Notably, the anti-tumor activity of resveratrol against NK cell lines was p53 independent as demonstrated by equal efficacy of resveratrol against NK cell lines pretreated with the p53 inhibitor Pifithrin-α. Immunoblot analysis to study intracellular signaling in resveratrol-treated cells showed suppression of constitutively active STAT3 in all four cell lines 24 hours after treatment. Remarkably, resveratrol inhibited JAK2 phosphorylation, but had no effect on other known upstream mediators of STAT3 activation such as PTEN and Tyk2 (Fig. 1b). Resveratrol induced robust G1 cell cycle arrest and down-regulation of two anti-apoptotic proteins, Mcl-1 and survivin, both of which are downstream effectors of the STAT-3 pathway. Furthermore, resveratrol enhanced the pro-apoptotic and anti-proliferative activities of L-asparaginase against NKL and NK-92 cells by 32% and 126% respectively. These data indicate that resveratrol possesses a potent anti-tumor effect via inactivation of the STAT3 pathway in malignant NK cells. These mechanistic findings suggest that resveratrol may have therapeutic potential against NK cell malignancies. Our finding that resveratrol is a bonafide JAK2 inhibitor extends its therapeutic potential to other diseases with deregulated JAK2 signaling. Disclosures: No relevant conflicts of interest to declare.