Objective: Although PU.1/Spi1 is known as a master regulator for macrophage development and function, we have reported previously that it is also expressed in adipocytes and is transcriptionally induced in obesity. Here, we investigated the role of adipocyte PU.1 in the development of the age-associated metabolic syndrome.Methods: We generated mice with adipocyte-specific PU.1 knockout, assessed metabolic changes in young and older adult PU.1fl/fl (control) and AdipoqCre PU.1fl/fl (aPU.1KO) mice, including body weight, body composition, energy expenditure, and glucose homeostasis. We also performed transcriptional analyses using RNA-Sequencing of adipocytes from these mice.Results: aPU.1KO mice have elevated energy expenditure at a young age and decreased adiposity and increased insulin sensitivity in later life. Corroborating these observations, transcriptional network analysis indicated the existence of validated, adipocyte PU.1-modulated regulatory hubs that direct inflammatory and thermogenic gene expression programs.Conclusion: Our data provide evidence for a previously uncharacterized role of PU.1 in the development of age-associated obesity and insulin resistance.

Obesity is associated with low-grade chronic inflammation, and metabolic dysregulation. Evidence shows that chronic inflammation inhibits protective immunity mediated by CD4+ T cells. Additionally, obesity-induced inflammation affects prostate cancer progression. However, the effect of obesity on CD4+ T-cell- response to prostate cancer is not well understood. To investigate whether obesity induces changes in CD4+ T cell cytokine profile, cytokine expression was measured in splenic CD4+ T-cells from 10-week-old male C57Bl/6 mice exposed to conditioned media (CM) from macrophages grown in sera from obese subjects. Additionally, expression levels of key regulators of Epithelial-Mesenchymal Transition (EMT) were measure in prostate cancer epithelial cells exposed to conditioned media from obesity-modified T-cells. Cell migration and invasion was measured in prostate cancer epithelial cells exposed to CM from obesity-modified CD4+ T-cells. Obesity suppressed the expression of IFNγ and IL-2 in CD4+ T-cells but up-regulated the expression of IL-6. Prostate epithelial cancer cells exposed to conditioned media from obesity-modified T cell increased the expression of EMT markers and showed a higher invasive and migratory capacity.

Abstract Background: Prostate cancer remains the second leading cause of cancer related deaths in men and obesity has been shown to severely impact its prognosis. Previous studies in our laboratory have shown that CD4+ T cells polarized under obese conditions are prostate cancer promoting and that this is due in part to suppression of IFNγ and induction of IL-6 in the T cells. Objective: The goal of this study is to determine the role of IL-10 in mediating the impact of obese conditions on T cell phenotype. Methods: A series of in vitro studies were conducted to evaluate the effects of obesity on T cell function and polarization. Splenic CD4+ T-cells from 10-week-old, male, C57BI/6 mice were isolated and stimulated with anti-CD3+/CD28+ antibodies under obese conditions. Gene expression was measured using qPCR and AKT phosphorylation was measured using Western blot analysis. In vivo T-cell function was compared between obese and non-obese male mice. Deletion assays were used to determine the role of IL-10 in mediating the effects observed on T cell phenotype in response to obese conditions. Results: For both the in vitro and in vivo studies, obese conditions suppressed the expression of Th1 biomarkers (IFNγ, IL-2, and T-bet) and upregulated the expression of Th2 biomarkers (IL-6, IL-10, and GATA-3) in CD4+ T cells. Likewise, obese conditions in vitro and in vivo were associated with decreased AKT phosphorylation. Studies are on-going to determine whether inhibiting the IL-10 pathway restores proper T cell activation under obese conditions. Conclusion: Obesity induces T cell dysfunction characterized by a Th1 to Th2 phenotypic shift, resulting in a pro-tumorigenic phenotype, potentially mediated by paracrine IL-10 signaling. These results are important in understanding the mechanisms by which obesity promotes a more aggressive disease, thereby identifying potential targets for improving outcomes. The results from these studies will provide a better understanding of the role of IL-10 as a novel target for preventing prostate cancer in obese subjects and improving overall survival. Citation Format: Margaret Kappel, Michael Adkison, Alejandra De Angulo, Peyton Travis, Brittany Harlow, Christopher Jolly, Linda deGraffenried. The role of IL-10 in obesity-induced T-cell polarization in the prostate tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2793.

Obese men are at greater risk of developing an aggressive form of prostate cancer. The T cell population from obese subjects have shown signatures of cell senescence including shortened telomeres and elevated production of proinflammatory cytokines. The objective of our studies is to better understand the cellular and molecular basis for altered T cell anti-prostate tumor immunity in obesity, and whether this altered immune function contributes to the worse outcomes observed in obese patients. To investigate whether obesity induces a pro-inflammatory senescent phenotype in CD4+ T cells, splenic CD4+ T-cells from 10-week-old male C57Bl/6 mice were isolated and, either pre-stimulated or co-stimulated with anti CD3/CD28 and conditioned media (CM) from macrophages stimulated under obese or normo-weight conditions. After 24 hours conditioned media from these CD4+ T cells was collected, and their relative cytokine expression profile was analyzed. The CD28 receptor is expressed on CD4+ T cells and provides co-stimulatory signals required for T cell activation. CD28 activation results in phosphorylation of Akt. Lack of Akt phosphorylation in CD4+ T cells after CD3+/CD28+ stimulation indicates CD4+ T cell dysfunction and senescence. Fresh murine splenic CD4+ T cells were isolated and stimulated with anti CD3/CD28 under obese or control conditions. Akt1 phosphorylation levels were assessed using Western blot analysis. To determine if obese conditions modify the impact of CD4+ T cell dysfunction on prostate cancer cell signal transduction pathways, prostate cancer epithelial cells were exposed to obese or control CD4+ T-cells CM for 30 minutes and western blot analysis was used to evaluate the modulation of key pro- and anti-tumorigenic pathways. Matrigel Invasion chambers were used to determine if obesity-modified CD4+ T cell CM impacts the invasive capacity of prostate epithelial cells. Obese conditions suppressed the expression of IFN-γ and IL-2 in CD4+ T-cells. Data also suggests that T cells exposed to obese conditions suppress STAT1 activation in prostate cancer cells. Additionally, obese conditions up-regulated the expression of IL-6 and IL-10 in CD4+ T-cells when compared to control. Importantly our data also suggests that T cells exposed to obese conditions induce STAT3 activation in prostate cancer cells. Preliminary data also suggests that obesity-modified CD4+ T cell CM impacts the invasive capacity of prostate cancer cells. Obesity may induce CD4+ T cell dysfunction, limiting their tumor suppressing activity. Findings form this study could substantially advance fundamental understanding of the ways in which obesity alters T cells, thereby promoting uncontrolled tumor growth and mortality from prostate cancer. This study could also provide a framework for future translational research aimed at improving the effectiveness of therapies in obese patients. Citation Format: Alejandra De Angulo, Peyton Travis, Christopher Jolly, Linda deGraffenried. Obesity-induced T cell senescence contributes to prostate cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 61.

Event Abstract Back to Event Dysregulation of IL-17 Production in Aged T-Lymphocytes Promotes Pro-tumorigenic Changes in Prostate Epithelial Cells Alejandra De Angulo1*, Robert Faris1, Christopher Jolly1 and Linda DeGraffenried1 1 University of Texas at Austin, United States Background: Prostate cancer represents 8% of all new cancer cases world-wide and is the second leading cause of cancer death in men. Incidence is closely associated with aging, but the mechanisms by which aging promotes prostate cancer development are unclear, limiting the development of effective preventive interventions. Aging is accompanied by immune dysfunction and a progressive increase in pro-inflammatory cytokines, including Interleukin 17 (IL-17). Aging-associated increases in circulating IL-17 promotes pro-inflammatory signaling in prostate epithelial cells, possibly contributing to tumorigenesis. Methods: Serum and splenic CD4+ T-lymphocytes from young T cell aging-mimic mice as well as young and aged wild-type mice were collected. Surface markers and intracellular levels of IL- 17, IFN-γ and IL-4 in isolated CD4+ T-cells were measured using flow-cytometry. shRNA was used to knock down the IL-17 receptor in LNCaP prostate cancer cells and RWPE-1 non-transformed prostate epithelial cells, which were then exposed to mouse sera or conditioned media from stimulated T-lymphocytes. NF-kB and STAT3 activation, NF-kB and STAT3 target gene expression, and epithelial cell transformation were all measured in prostate epithelial cells. Results: T cells from aging and aging-mimic mice secrete elevated levels of IL-17, possibly due to an imbalance in the TH17/TH1 and TH17/TH2 cell ratios. T-lymphocyte-secreted IL-17 from aging-mimic mice induced NF-kB and STAT3 activity and target gene expression in LNCaP and RWPE-1 cells. Importantly, IL-17 also promoted pro-tumorigenic changes in RWPE-1 cells, leading to a pre-transformed phenotype. Inhibition of IL-17 signaling blocked age-induced changes in both the cancer and non-transformed prostate epithelial cells. Conclusion: Aging of the immune system is associated with increased levels of IL-17, which stimulates the pro-inflammatory NF-kB and STAT3 pathways in prostate epithelial cells and activates programming associated with cellular transformation. Inhibition of IL-17 signaling blocks the pro-tumorigenic activity induced by aging T cells. These findings provide evidence that the dysregulation of IL-17 production in aged T-cells may directly contribute to the increased risk for prostate cancer. Future novel immunotherapies for prostate cancer prevention could target the increase production of IL-17 associated with aging References 1. De Angulo, A., et al.Age-related alterations in T-lymphocytes modulate key pathways in prostate tumorigenesis. Prostate, 2013. 73(8): p. 855-64. 2. Schmitt, V., L. Rink, and P. Uciechowski, The Th17/Treg balance is disturbed during aging. Exp Gerontol, 2013. 48(12): p. 1379-86. 3. Zhang, Q., et al., Interleukin-17 promotes formation and growth of prostate adenocarcinoma in mouse models. Cancer Res, 2012. 72(10): p. 2589-99 4. Zhang, Q., et al., Interleukin-17 promotes development of castration-resistant prostate cancer potentially through creating an immunotolerant and pro-angiogenic tumor microenvironment. Prostate, 2014. 74(8): p. 869-79. 5. You, Z., et al., Interleukin-17 receptor-like gene is a novel antiapoptotic gene highly expressed in androgen-independent prostate cancer. Cancer Res, 2006. 66(1): p. 175-83. 6. Sfanos, K.S., et al., Phenotypic analysis of prostate-infiltrating lymphocytes reveals TH17 and Treg skewing. Clin Cancer Res, 2008. 14(11): p. 3254-61. Keywords: Interleukin 17, T-Lymphocytes, NF-kappa B, STAT3 Transcription Factor, prostate cancer, Aging Conference: IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología, Medellin, Colombia, 13 Oct - 16 Oct, 2015. Presentation Type: Poster Presentation Topic: Tumor immunology Citation: De Angulo A, Faris R, Jolly C and DeGraffenried L (2015). Dysregulation of IL-17 Production in Aged T-Lymphocytes Promotes Pro-tumorigenic Changes in Prostate Epithelial Cells. Front. Immunol. Conference Abstract: IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología. doi: 10.3389/conf.fimmu.2015.05.00053 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 07 Apr 2015; Published Online: 14 Sep 2015. * Correspondence: Dr. Alejandra De Angulo, University of Texas at Austin, Austin, TX, United States, alejandra.deangulo@utexas.edu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Alejandra De Angulo Robert Faris Christopher Jolly Linda DeGraffenried Google Alejandra De Angulo Robert Faris Christopher Jolly Linda DeGraffenried Google Scholar Alejandra De Angulo Robert Faris Christopher Jolly Linda DeGraffenried PubMed Alejandra De Angulo Robert Faris Christopher Jolly Linda DeGraffenried Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

BackgroundA close relationship between aging, inflammation, and prostate cancer is widely accepted. Aging is accompanied by a progressive increase in pro-inflammatory cytokines, including interleukin 17 (IL-17), a key pro-inflammatory cytokine that becomes dysregulated with age. However, the contribution of IL-17 to age-related prostate tumorigenesis remains unclear. The aim of this study was to investigate the role of age-related IL-17 dysregulation in prostate tumorigenesis.MethodsSerum and splenic T-lymphocytes from young GPAT-1 knock-out aging-mimic T cell mice as well as young and aged wild-type mice were collected. shRNA was used to knock down the IL-17 receptor in LNCaP prostate cancer cells and RWPE-1 non-transformed prostate epithelial cells, which were then exposed to the mouse sera or conditioned media from stimulated T-lymphocytes. NF-B activation, NF-B target gene expression, and cell proliferation were all measured in these cells by luciferase assay, qPCR, Western blot analysis, and MTT assay, respectively.ResultsT-lymphocyte-secreted IL-17 from aging-mimic mice induced NF-B activity and target gene expression in LNCaP and RWPE-1 cells. It also promoted proliferation of these cells.ConclusionAging-mimic T cell mice produce increased levels of IL-17, which stimulates the pro-inflammatory NF-B pathway in prostate epithelial cells. NF-B increases inflammation, carcinogenesis and metastatic potential in the prostate. These findings provide evidence that the dysregulation of cytokine production seen in aged T cells may directly contribute to the increased risk for prostate cancer in the elderly. Prostate 75: 449-462, 2015. (c) 2015 Wiley Periodicals, Inc.

Glycerol-3-phosphate acyltransferase-1 is the first rate limiting step in de novo glycerophospholipid synthesis. We have previously demonstrated that GPAT-1 deletion can significantly alter T cell function resulting in a T cell phenotype similar to that seen in aging. Recent studies have suggested that changes in the metabolic profile of T cells are responsible for defining specific effector functions and T cell subsets. Therefore, we determined whether T cell dysfunction in GPAT-1 (-/-) CD4(+) T cells could be explained by changes in cellular metabolism. We show here for the first time that GPAT-1 (-/-) CD4(+) T cells exhibit several key metabolic defects. Striking decreases in both the oxygen consumption rate (OCR) and the extracellular acidification rate (ECAR) were observed in GPAT-1 (-/-) CD4(+) T cells following CD3/CD28 stimulation indicating an inherent cellular defect in energy production. In addition, the spare respiratory capacity (SRC) of GPAT-1 (-/-) CD4+ T cells, a key indicator of their ability to cope with mitochondrial stress was significantly decreased. We also observed a significant reduction in mitochondrial membrane potential in GPAT-1 (-/-) CD4(+) T cells compared to their WT counterparts, indicating that GPAT-1 deficiency results in altered or dysfunctional mitochondria. These data demonstrate that deletion of GPAT-1 can dramatically alter total cellular metabolism under conditions of increased energy demand. Furthermore, altered metabolic response following stimulation may be the defining mechanism underlying T cell dysfunction in GPAT-1 (-/-) CD4(+) T cells. Taken together, these results indicate that GPAT-1 is essential for the response to the increased metabolic demands associated with T cell activation.

BACKGROUND The primary risk for prostate cancer is aging, often associated with inflammation. Evidence implicates progressive age-related immune dysfunction with increased prostate cancer incidence and mortality. The aged T-cell response is characterized by increased production of pro-inflammatory cytokines, which could significantly contribute to prostate tumorigenesis through induction of key inflammation-mediated pro-survival factors. METHODS T-cell function of the young (<6 month-old) glycerol-3-phosphate acyltransferase-1 (GPAT-1) knock-out mouse mimics many of the hallmarks observed in an aged (>22-month-old) mouse. Serum and splenic T-lymphocytes from young GPAT-1/ (6 months) and aged wild type (22 months) mice were collected for in vitro studies, including a cytokine immunoarray for serum cytokine levels, luciferase assays for NF-B activation and Western blot analyses for protein expression. RESULTS The T-cell cytokine profile of the GPAT-1/ mice mirrored that observed in aged wild type mice, including higher expression levels of IL-17. Serum- and T-cell-derived factors induced NF-B activity in normal, non-transformed and prostate cancer epithelial cells, correlating with re-localization of NF-B and increased protein expression of downstream targets of NF-B. CONCLUSION The aging and aging-mimic mice produced circulating factors that induce pro-inflammatory pathways in prostate cells, most notably NF-B. These findings provide evidence that an aged T-cell may directly contribute to the increased risk for prostate cancer in the elderly and establish that the GPAT-1/ model, which mimics many of the characteristics of an aged immune system, is a viable tool for investigating this novel area of cancer risk. Prostate 73: 855864, 2013. (c) 2013 Wiley Periodicals, Inc.

Prostate cancer risk is determined largely by age. While some studies have suggested that the slow growing rate of prostate cancer is responsible for the link between aging and prostate cancer, the role of aging T-cells in prostate cancer development remains largely unknown. The objective of this study is to determine how age-related changes in T-lymphocyte function contribute to prostate tumorigenesis. We hypothesized that age related changes in T-lymphocyte function, to a pro-inflammatory phenotype may promote prostate cancer development. For our studies we used the GPAT-1 KO mice model. The GPAT-1 KO mice have T-lymphocytes that mimic aged T-cells. Preliminary data shows that, in agreement with what has been seen in human aged T-cells, aging-mimic GPAT-1 KO T-lymphocytes generate more pro-inflammatory cytokines than T-lymphocytes from age matched wild-type mice. These cytokines turn on inflammatory pathways that stimulate proliferation, tissue disruption and tumor growth. A cytokine array was used to detect the relative protein levels of 40 different cytokines and chemokines in the sera from either young GPAT-1 knockout (KO), young wild-type (WT) or old mice (22 months). We observed a shift to a more pro-inflammatory cytokine profile in the GPAT-1 KO and old mice. Many of the same cytokines that were elevated in the sera from the old and GPAT-1 KO mice, such as IL-17 and IL-6, are also implicated in promoting prostate tumorigenesis primarily through induction of other inflammatory cytokines and transcription factors such as, NF-kB and STAT3. The objective of this study was to determine if the increased levels of pro-inflammatory cytokines in the sera from the GPAT-1 KO mice correlates to a more malignant phenotype in prostate epithelial cells. Malignancy in epithelial cells was assessed by the following phenotypical changes: Immortalization, anchorage independence, proliferation and invasiveness. Exposure to sera from the GPAT-1 KO mice promoted phenotypical changes in epithelial cells that are indicative of early tumorigenesis. Taken together our data suggests that pro-inflammatory cytokines in the sera from mice with aging T-lymphocytes may turn on pro-inflammatory pathways that contribute to a more malignant phenotype in prostate epithelial cells. This is the first study to use the GPAT-1 KO mice model to explore the relationship between aging T-lymphocytes, inflammation and prostate tumorigenesis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3553. doi:1538-7445.AM2012-3553

Abstract Background: Age is the single greatest risk factor for prostate cancer development. As men pass age 40, their chance of developing prostate cancer increases exponentially each year. The mechanism by which age influences the risk of prostate cancer remains largely unknown, hindering the development of effective preventive approaches. While local events are known to contribute to age-related prostate tumorigenesis, the role of the aging immune system in prostate cancer development has been largely understudied. The aging process affects the adaptive cell-mediated immune response and at the same time there is a shift to a more inflammatory cytokine profile. Recent studies have shown that age-related enhancements of Th17 might contribute to changes in immune function and prostate cancer development. The age-related shift in T-lymphocyte cytokine profile and the enhancement of TH17 generation might be significant contributing factors to the association between age and prostate tumorigenesis. Methods: For our studies we have been using a unique animal model, the glycerol-3-phosphate acyltransferase-1 (GPAT-1) knock out mouse. The GPAT-1 KO young mouse T lymphocyte phenotype mimics old (> 18 m) wild type mice. Preliminary data suggested that cytokines secreted from the GPAT-KO aging mimic T-lymphocytes can promote activation of the STAT3/NF-kB pathways. In order to test if IL-17 was responsible for the strong induction of NF-kB and STAT3, we repeated previous experiments that measured the activation of NF-kB and STAT3, but supplemented the experimental sera and conditioned media with depleting antibodies to IL-17. Successful inhibition of the NF-kB and STAT3 pathways was assessed by Western Blot analysis. Conclusions: Depleting IL-17 in the GPAT-1 KO sera and conditioned media decreased NF-kB activity to the levels of the cells treated with wild-type sera. Sera from the old and GPAT-1 KO mice induced activation of downstream targets of IL-17. These results suggest that circulating factors from the GPAT-1 KO mice mimic those in aging mice and strongly induced signaling pathways involve in the development of prostate cancer. The age-related enhancements of IL-17 may play a key role in prostate cancer initiation and progression. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5577. doi:10.1158/1538-7445.AM2011-5577

Abstract As men age past 40, their chance of developing prostate cancer increases exponentially each year. The mechanism by which age influences the risk of prostate cancer remains largely unknown, hindering the development of effective preventive interventions. As one ages, there are significant changes in immune response regulation and outcome. This includes an age-related loss of appropriate T cell function. This loss of appropriate T cell function is associated with a change from anti-inflammatory to pro-inflammatory cytokine production as well as decreased appropriate immune surveillance. We hypothesize that one mechanism by which age increases the risk for prostate cancer may be through these alterations in immune response by the T cells. To date, the specific contribution of an aging immune system to prostate cancer initiation and progression has been difficult to determine because of the lack of appropriate model systems to evaluate this phenomena. We have recently begun studies investigating the contribution of an aging T cell population to prostate cancer progression using a highly unique and innovative animal model, the glycerol-3-phosphate acyltransferase-1 (GPAT-1) knock out mouse. The GPAT-1 KO young mouse has a T cell phenotype which mimics old wild type mice. We have recently initiated a series of in vitro studies to evaluate the effects of exposure of sera from GPAT KO mice on pro-inflammatory signaling in LNCaP prostate cancer cells. Exposure to sera from young GPAT KO mice resulted in strong induction of NF-κB activity in LNCaP cells compared to both the sera from the age-controlled wild type mice and the sera-free conditions. We are currently evaluating the significance of this NF-κB induction as well as the induction of other pro-inflammatory pathways by the GPAT KO mouse sera in promoting a more aggressive phenotype in our cancer cells. Additionally, LNCaP cells exposed to conditioned media from stimulated T-cells from the GPAT KO mice demonstrated induction of Akt phosphorylation. Previous studies have shown an important role for NF-kB and Akt in promoting inflammation and survival in prostate cancer cells. Currently, we are evaluating how GPAT KO mouse sera might promote a more aggressive phenotype in LNCaP prostate cancer cells. These studies would be the first to demonstrate a specific modulation of prostate cancer signaling by sera from mice that directly reflect an aging T cell phenotype suggesting that age-related loss of appropriate T cell function might contribute to the increased risk for prostate cancer in the elderly. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2472.