Citations

AbstractHematopoietic stem cell (HSC) self‐renewal is regulated by osteoblast and/or endothelial cells within the hematopoietic niche. However, the true identity of the supporting cells and the nature of the secreted factors remain uncertain. We developed a novel mouse model and analyzed whether circulating human peripheral hematopoietic lineage negative/AP+ (lin−/AP+) cells support hematopoiesis in vivo. Thus, immunocompromised (Rag) mice expressing thymidine kinase (Tk) under the control of the 3.6Col1α1 promoter (Tk‐Rag) were treated with ganciclovir, resulting in osteoblast progenitor cell ablation and subsequent loss of hematopoiesis (evaluated by measuring mouse Ter119+ erythroid cells). Following hematopoietic cell depletion, human bone marrow‐derived marrow stromal cells (MSCs) or lin−/AP+ cells were infused into Tk‐Rag mice and compared with saline infusions. Ganciclovir significantly reduced (7.4‐fold) Ter119+ cells in the bone marrow of Tk‐Rag mice compared to saline injections. Infusion of either MSCs or lin−/AP+ cells into ganciclovir‐treated mice resulted in a 3.3‐fold and 2.7‐fold increase (P < 0.01), respectively, in Ter119+ cells compared to mice receiving saline. Relative to lin−/AP− cells, lin−/AP+ cells expressed high levels of mesenchymal, endothelial, and hematopoiesis supporting genes. Thus, human peripheral blood lin−/AP+ cells represent a novel cell type capable of supporting hematopoiesis in a manner comparable to MSCs. J. Cell. Biochem. 116: 58–66, 2015. © 2014 Wiley Periodicals, Inc.

AbstractObjective The high level of stress associated with caring for others with medical conditions has been recognized for some time. Reducing caregiver stress can improve caregiver quality of life as well as improve the care they provide to loved ones. This systematic review assesses the effectiveness of internet-based interventions to decrease caregiver stress.Materials and methods A comprehensive literature search of Ovid MEDLINE (1946–2013), Embase (1988–2013), PsycINFO (1987–2013), and CINAHL was conducted using terms related to caregiver and internet-based interventions. Internet-based interventions involving informal caregivers in an open-label or randomized controlled trial setting were included. A pair of raters independently reviewed all published abstracts. Data regarding participants, interventions, and outcomes were extracted and, for randomized trials, methodology quality was also reviewed.Results Eight open-label trials met the review criteria: three showed positive benefit in reducing caregiver stress, four were partially positive (some outcomes positive, others negative), and one was a negative study. Sixteen randomized trials met the review criteria: six showed positive benefit, five were partially positive, and five were negative. There were no clear patterns as to the variables (such as study duration and complexity of intervention) associated with better outcomes, although earlier studies typically had more negative outcomes.Discussion Internet-based interventions were mostly effective in reducing aspects of caregiver stress and improving their well-being. Further studies to assess outcomes for caregivers and their recipients’ health, different technology delivery methods, and the cost of such interventions are needed.

Abstract Ovarian cancer is the fifth leading cause of cancer death among women in the United States. Despite advances in treatment for other cancers, ovarian cancer patient prognosis has not improved significantly since the advent of platinum-based chemotherapy. It is a highly heterogeneous disease with at least four distinct histological subtypes: serous, mucinous, endometrioid, and clear cell. However, no subtype- or biomarker-specific therapies are approved for ovarian cancer. Identifying targeted therapies is crucial to improving clinical outcomes. Specific therapies will likely vary based on tumor histology and biomarkers. Expression of estrogen receptor alpha (ER) is a well established biomarker of response to endocrine therapies in breast cancer. Despite the success of anti-estrogen therapies in treating breast cancer, little attention has been given to their potential in ovarian cancer. However, epidemiologic studies show that approximately 70% of epithelial ovarian tumors express ER, particularly serous and endometrioid subtypes, and that estrogen plays a role in ovarian cancer etiology. Further, clinical data suggest that a subset of ovarian cancer patients can be successfully treated with endocrine therapy. We hypothesize that some ovarian tumors require estrogen for growth and that predictive biomarkers will identify patients who will benefit from endocrine therapy. Using cell line models, we examined the response of ovarian cancer cells to estrogen (E2) and anti-estrogens with regard to gene expression and cell proliferation. E2 induces expression of canonical ER target genes (e.g. GREB1) in these cells, suggesting that they are estrogen-responsive. We observe that 4-hydroxytamoxifen and fulvestrant inhibit growth of ovarian cancer cells including the chemo-resistant line PEO4. Gene expression microarray studies identified additional E2-regulated genes in ovarian cancer cells that may be utilized as biomarkers of ER activity. We are expanding our studies to in vivo models (human primary tumor xenografts) to evaluate the role of E2 in driving tumor progression and determine the efficacy of anti-estrogens in treating serous ovarian cancer. There is a critical need for targeted therapies in ovarian cancer. Our studies will determine if endocrine responsiveness correlates with gene expression of specific biomarkers, which may lead to the identification of predictive clinical markers of response to endocrine therapy. Citation Format: Courtney L. Andersen, Matthew J. Sikora, Paul Haluska, Steffi Oesterreich. Identifying biomarkers of estrogen response in models of serous ovarian cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 94. doi:10.1158/1538-7445.AM2013-94

Abstract Tumor infiltrating regulatory T cells (Tregs) appear to play a role in survival in a number of solid tumors, including ovarian cancer. We assessed genetic variation via tag SNPs (N=749) in 25 Treg associated genes (CD28, CTLA4, FOXP3, IDO1, IL10, IL10RA, IL15, 1L17RA, IL23A, IL23R, IL2RA, IL6, IL6R, IL8, LGALS1, LGALS9, MAP3K8, STAT5A, STAT5B, TGFB1, TGFB2, TGFB3, TGFBR1, TGRBR2, and TGFBR3) in relation to ovarian cancer survival. SNPs were selected based on r2 ≥ 0.8, MAF ≥ 0.05, 5 kb upstream and downstream from the gene. We had SNP and outcome data for ovarian cancer from 10,084 combined ovarian cancer cases, including 5,248 high grade serous, 1,452 endometrioid, 661 mucinous, and 795 clear cell cases of European descent across multiple studies from the Ovarian Cancer Association Consortium (OCAC). Cox proportional hazards regression modeling was used to estimate per-allele hazard ratios (HRs) and 95% confidence intervals (CIs) for associations with overall survival, adjusting for age at diagnosis, population substructure PCs, study site, histology (for the all case analysis), tumor stage, tumor grade and oral contraceptive use. The strongest associations were found for endometrioid histologic subtype and IL2RA SNPs (maximum r2=0.67) rs11256497 [HR=1.42, 95% CI: 1.22-1.64; p=5.7 x 10−6], rs791587 [HR=1.36, 95% CI: 1.17-1.57; p=6.2 x 10−5], rs2476491 [HR=1.40, 95% CI: 1.19-1.64; p=5.6 x 10−5], and rs10795763 [HR=1.35, 95% CI: 1.17-1.57; p=7.9 x 10−5], as well as clear cell ovarian cancer and CTLA4 SNP rs231775 [HR=0.67, 95% CI: 0.54-0.82; p=9.3 x 10−5]. No other associations were observed at p&lt;1 x 10−4. Overall, this study provided evidence that variation in genes related to Tregs, particularly IL2RA and CTLA4, influence ovarian cancer survival. Citation Format: Bridget Charbonneau, Keith Knutson, Robert A. Vierkant, Julie M. Cunningham, Zachary C. Fogarty, Kimberly R. Kalli, Matthew J. Maurer, Ann L. Oberg, Brooke L. Fridley, Claudia C. Preston, Paul D.P. Pharoah, Susan J. Ramus, Catherine M. Phelan, Kunle Odunsi, Kirsten Moysich, Ellen L. Goode. Polymorphisms in regulatory T cell related genes and ovarian cancer survival. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4844. doi:10.1158/1538-7445.AM2013-4844

Abstract Background: Ovarian cancer (OC) is the leading cause of gynecologic cancer death in the United States. Survival in advanced OC is highly variable and is impacted by the endogenous anti-tumor immune response. Toll-like receptors (TLR) on tumor-infiltrating leukocytes can modulate anti-OC immunity. Heparan sulfate (HS) is an oligosaccharide component of the extracellular matrix that, when released, binds and triggers TLR-4 to induce production of multiple cytokines; HS is liberated from the extracellular matrix by heparanase, which is overexpressed by many ovarian cancers. We therefore explored the association between plasma levels of HS, heparanase, proinflammatory and immunosuppressive cytokines, and survival in patients with advanced OC. Methods: We designed a two stage cohort study of plasma biomarkers in ovarian cancer patients. For the pilot stage, we enrolled 100 patients with advanced OC (AJCC Stage III and IV), 50 patients with early stage OC (Stage I and II), and 50 patients with benign surgical conditions from the Mayo Ovarian SPORE Biospecimens Core. For the expansion stage, 257 patients with advanced OC were enrolled. Presurgical plasma samples were assayed for HS, heparanase, and multiple cytokines by ELISA and electrochemiluminescence. Differences in plasma levels between benign, early, and advanced OC patient groups were assessed using plate-adjusted linear models. Associations between plasma levels of HS and heparanase with overall survival in advanced OC patients were assessed by Cox proportional hazards models using a log2 transformation; total effects across stages were estimated using a meta-analysis approach. Results: In the pilot cohort, soluble HS levels were similar in control (mean +/ STD: 88.26 +/- 32.23), early stage (93.91 +/- 37.77), and advanced OC (93.55 +/- 40.44), with similar values observed in the expansion cohort (129.29 +/- 95.64). There was no correlation between heparanase and HS levels. Higher levels of HS were associated with improved overall survival (log2 HR = 0.62, p = 0.042; HR = 0.84, p = 0.014; HR = 0.82, 95% CI: 0.71-0.93, p = 0.003 for the pilot, expansion, and combined sets, respectively). There was no association between heparanase levels and overall survival (all p &gt; 0.45). Conclusions: Neither soluble HS nor heparanase were elevated in presurgical OC patients relative to controls; however, higher levels of HS were associated with improved survival in patients with advanced OC. These results suggest that HS may play a key role in modulating the innate immune response to OC. Citation Format: Matthew S. Block, Matthew J. Maurer, Krista M. Goergen, Courtney L. Erskine, Marshall D. Behrens, Kimberly R. Kalli, Keith L. Knutson. Higher plasma levels of heparan sulfate are associated with improved survival of patients with advanced ovarian cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3476. doi:10.1158/1538-7445.AM2013-3476

Abstract Background: Ovarian cancer (OC) is commonly associated with ascites and predominantly remains in the peritoneal cavity. Soluble factors present in ascites such as growth factors and cytokines likely play a key role in local tumor behavior including stage, invasiveness and resectability and survival. Members of IL-12 family (IL-12p70, IL-23, IL-27 and IL-35) play a critical role in the regulation of inflammatory response. However, their role as local factors in ovarian cancer growth has not been well studied. We hypothesized that that the level of these important immunomodulatory and pro-inflammatory cytokines are associated with the clinical behavior of OC. Methods: Ascites fluid was collected and analyzed from women undergoing maximal debulking surgery for presumed OC, and relevant clinical endpoints were abstracted. Concentrations of IL-12 related cytokines (IL-23, IL-27, IL-35, and IL-12p70) were assessed by ELISA. Cytokines were also assessed on matching pre-surgery plasma samples. Cytokine concentrations between matched ascites and plasma samples were examined using scatter plots and Spearman correlations. Associations between ascites concentrations and clinical characteristics were assessed using Wilcoxon rank-sum tests. Associations between ascites levels with overall survival in newly diagnosed invasive OC patients were assessed by Cox proportional hazards models using a log2 transformation. Results: Ascites fluid was analyzed from 20 patients with primary epithelial OC including 18 serous histology, 19 patients with stage III or IV disease: 16 (80%) were optimally debulked. We observed that higher levels of IL-12p70 were associated with locally invasive disease including higher rates of sub-optimal debulking (p=0.0074) and stage IV disease (p=0.012). For other IL-12 family members we observed similar associations between higher cytokine levels and suboptimal disease and stage IV disease including IL-23 (p=0.73, p=0.17), IL-27 (p=0.041, p=0.018), and IL35 (p=0.12, p=0.11), respectively. In addition, increase in IL-23 was associated with worse overall survival (log2 HR = 1.85, p=0.031). Conclusions: Higher levels of IL-12 family of cytokines in ascitic fluid are associated with several surrogates of biological aggressiveness in OC including stage, resectability and survival. Given the importance of local tumor environment on clinical behavior, a detailed understanding of IL-12 pathway may help in developing novel therapeutic strategies to treat OC. Citation Format: Eati Basal, Mathew S. Block, Matthew J. Maurer, Krista Goergen, Courtney L. Erskine, Marshall Behrens, Kimberly R. Kalli, Keith Knutson, William Cliby. Levels of IL-12 family cytokines in ascitic fluid are associated with extent and resectability of ovarian cancer and correlate with overall survival. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1434. doi:10.1158/1538-7445.AM2013-1434

Abstract NSC 161128 is a phenylurea thiocarbamate that has shown differential activity in the NCI60 anticancer screen and in xenograft models. The best in vivo activity was observed against the DU-145 prostate cell line. Previous studies conducted by DTP/NCI demonstrated that NSC 161128 is rapidly metabolized after administration to mice or rats, with the major detected metabolite being N-methyl-N’-phenylurea (M10). The other putative metabolite(s) arising from cleavage of the N-S bond, dimethyldithiocarbamate (M85) and/or its disulfide [bis(dimethyldithiocarbamoyl)disulfide, M17] were not detected in animal plasma using available methodologies. NSC 161128 was also shown to be rapidly degraded in aqueous solution at low and high pH, forming M10 and M17. As part of the preclinical evaluation of NSC 161128, we developed a sensitive, specific LC/MS/MS method to quantitate NSC 161128, M10 and M17 in biological samples, explored the stability of NSC 161128 in cell culture medium, and characterized the pharmacokinetics and in vivo metabolism of NSC 161128 following i.p. administration to athymic nude mice. We developed and validated suitable positive ionization lc/ms/ms methodology for measuring NSC 161128, M10 and M17. The assay utilized liquid extraction with isopropanol:1-chlorobutane, 1:9 (v/v) and MS/MS detection with the 270&gt;177, 151&gt;94 and 241&gt;120 transitions for NSC 161128, M10 and M17, respectively. Baseline separation was achieved on an Atlantis T3 column under a mobile phase consisting of 40:60 methanol:water containing 0.1% formic acid. Standard curves were linear over the concentration range 1.0-1000 ng/mL with a lower limit of detection of 1.0 ng/mL. The parent compound was stable under acidic and neutral conditions, but some degradation (&lt;10%) was detected under basic conditions. NSC 161128 disappearance (∼20%) was also detected during 24-h incubation in RPMI medium +/- FBS. M10 was a minor product in those same incubations, suggesting that reactive metabolites are not formed in cell culture medium. Rapid degradation of NSC161128 and concomitant formation of M10 was observed in plasma at room temperature. While degradation was reduced by incubation on ice, substantial loss of drug (&gt;40%) was noted within 30 minutes, and samples for pharmacokinetic studies were kept cold and processed quickly to prevent degradation. NSC 161128 pharmacokinetics were characterized in male CD1 mice after i.p. administration of a 200 mg/kg dose. The peak plasma concentration of 255 ng/ml was observed 5 minutes after drug administration and the plasma half-life was 138 min. The peak concentration of 14.7 μg/ml M10 was detected 15 min after the NSC 161128 dose and the half-life was 560 min. Analysis of the pharmacokinetics of the thiol metabolites will be presented. Supported by NCI contract N01-CM-2011-00014. Citation Format: Chad A. Walden, Joel M. Reid, Renee M. McGovern, Joseh M. Covey, Matthew M. Ames. LC/MS/MS assay and mouse pharmacokinetics of the phenylurea thiocarbamate NSC 161128. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4557. doi:10.1158/1538-7445.AM2013-4557

Abstract BACKGROUND: Biobanks collect and store biological samples and data from participants for use in research by multiple investigators. The Mayo Clinic Biobank was established by the Mayo Clinic Center for Individualized Medicine with a goal to support a wide array of health-related research studies at Mayo Clinic, including cancer studies. The Biobank is not focused on any particular disease, however, through annotation by questionnaires, the Mayo electronic medical record (EMR), and the Mayo Cancer Registry, it is a useful clinical research resource for cancer related studies. METHODS: Recruitment started in April 2009. Our goal is 50,000 enrollees. Eligible subjects are adult Mayo Clinic patients, U.S residents, and able to consent. Mailed invitations were sent to patients with appointments in certain Mayo Clinic departments. Each participant completed a health history questionnaire including a wide variety of health information as well as whether they or their 1st-degree relatives had ever been diagnosed with specific cancers. For self-reported cancers, subjects were asked to provide the age they were first diagnosed. Subjects also provided a blood sample and granted research access to clinically collected specimens and data from their Mayo Clinic record, at enrollment and into the future. All research projects utilizing samples from this resource are required to return to the Biobank the data generated from their projects. RESULTS: In the first three years, 21736 subjects consented to participate (30% response rate). Participants are 58% female, 95% White, and range in age from 18 to 97 with a median age of 62. At enrollment, 74% were residents in Minnesota, 6% in Iowa, 4% in Wisconsin, and 15% in the other states. Twelve percent reported being a current smoker, while 59% were never smokers. The most common self-reported cancers at baseline were non-melanoma skin cancer (n=2950), prostate cancer (n=1107), breast cancer (n=941), melanoma (n=692), colorectal cancer (n=296), cervical cancer (n=240), sarcoma (n=233), urinary/bladder (n=211), lymphoma (n=187), thyroid cancer (n=186), endometrial cancer (n=170), kidney cancer (n=163), lung cancer (n=145). Excluding non-melanoma skin cancer, 16188 (74%) (6516 male, 9672 female) self-reported being cancer free at baseline. Data will be presented on the frequency of incident cancers diagnosed since enrollment in the Biobank. To date, the Biobank has served as a source of controls for 17 cancer related studies on genetic susceptibility, including studies of the breast, colon, myeloma, brain, lung, pancreas, endometrium, and kidneys. CONCLUSIONS: The Mayo Clinic Biobank has quickly been established as a valuable resource for cancer researchers at Mayo Clinic - primarily as a source of controls for genetic studies. As the Biobank continues to grow in sample size and length of follow-up, it will also serve as a cohort study of cancer incidence embedded within an EMR of a large health organization. Citation Format: Janet E. Olson, Euijung Ryu, Kiley J. Johnson, Karen Maschke, Jody A. Morrisette, Mark Liebow, Paul Y. Takahashi, Ruchi G. Sharma, Kari S. Anderson, Matthew A. Hathcock, Jyotishman Pathak, Noralane M. Lindor, Timothy J. Beebe, Stephen N. Thibodeau, James R. Cerhan. The Mayo Clinic Biobank - A growing resource for cancer related studies. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2515. doi:10.1158/1538-7445.AM2013-2515

Abstract NSC 373981 is 1-ethanol, 2-nitro-naphtho[2,1-b]furan that has demonstrated activity in the NCI60 anticancer screen and in the mouse hollow-fiber assay. Growth inhibitory activity was also observed in several xenograft models. There is some evidence that antitumor activity is greater in vivo than would be predicted by the in vitro results, suggesting the possibility of activating metabolism. While the mechanism of anticancer activity is unknown, naphthofurans have been reported to modulate certain nuclear receptors. The purpose of this study was to characterize the plasma pharmacokinetics of NSC 373981 following i.v. and i.p. administration to athymic nude mice and to characterize potential metabolites of this compound in plasma, urine, and bile. We developed and validated suitable positive ionization LC/MS/MS methodology for measuring NSC 373981 that utilized liquid extraction with 1-chlorobutane and MS/MS detection with the 258&gt;212 and 244&gt;169 transitions, respectively, for NSC 373981 and NSC 329226, an analog that was used as the internal standard. Baseline separations were achieved on a Polaris C18 column under a mobile phase consisting of 70:30 methanol:water containing 0.1% formic acid. Standard curves were linear over the concentration range 2.5-1000 ng/mL with a lower limit of detection of 2.5 ng/mL. NSC 373981 was stable under acidic, neutral and basic conditions, as well as human and mouse plasma during 24-hr incubation at 37°C. NSC 373981 pharmacokinetics were characterized in male nu/nu mice after a 37.5 mg/kg i.v. dose or a 150 mg/kg i.p. dose. Plasma, bile, urine and feces were collected for 24-hr after drug administration. Following i.v. administration, the peak concentration was &gt; 5.9 μg/mL and the terminal elimination half-life was 1.2 hours. Volume of distribution and plasma clearance values were 694 ml and 390 mL/hr, respectively. Following i.p. administration, NSC373981 reached a peak concentration of 2.1 μg/mL at 15 minutes and the terminal elimination half-life was 36.8 hours. The i.p. bioavailability of NSC373981 was 58%. In vivo metabolite profiles were characterized in plasma, urine, feces, and bile. We detected several Phase I metabolites in these samples including those produced by reduction of the aromatic nitro group, oxidation of the primary alcohol moiety on the reduced metabolite, oxidation of the primary alcohol moiety, benzylic hydroxylation, hydroxylation and dihydrodiol formation on the naphthalene moiety, and Phase II metabolites including glucuronidation of the carboxylic acids and a glutathione metabolite of hydroxylated NSC 373981. Supported by NCI contract N01-CM-2011-00014. Citation Format: Joel M. Reid, Sarah A. Buhrow, Renee M. McGovern, Joseph M. Covey, Matthew M. Ames. Pharmacokinetics and in vivo metabolism of the naphthofuran NSC373981 in male nu/nu mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2462. doi:10.1158/1538-7445.AM2013-2462