This file contains supplementary figures S1-S4. These illustrate correlation of ploidy from chromosome 17 FSIH versus 6-chromosome FISH, chromosome spreads of cell lines, additional dose-response curves, and data demonstrating DPBQ does not operate by binding DNA or by inhibiting topoisomerase II.

Supplemental Materials and Methods

Supplemental Table 4. Clinical characteristics of patients prior to surgery and PDX establishment

Supplemental Table 2. Clinical parameters and median progression-free survival in relation to AXL expression among patients with HNSCC

Supplemental Table 3. Fractional cell proliferation analysis for drug combination synergy

Supplemental Table 1. Clinical information describing HNSCC cohort

Inflammation, and the organization of collagen in the breast tumor microenvironment, is an important mediator of breast tumor progression. However, a direct link between markers of inflammation, collagen organization, and patient outcome has yet to be established. A tumor microarray of 371 invasive breast carcinoma biopsy specimens was analyzed for expression of inflammatory markers, including cyclooxygenase 2 (COX-2), macrophages, and several collagen features in the tumor nest (TN) or the tumor-associated stroma (TS). The tumor microarray cohort included females, aged 18 to 80 years, with a median follow-up of 8.4 years. High expression of COX-2 (TN), CD68 (TS), and CD163 (TN and TS) predicted worse patient overall survival (OS). This notion was strengthened by the finding from the multivariate analysis that high numbers of CD163+ macrophages in the TS is an independent prognostic factor. Overall collagen deposition was associated with high stromal expression of COX-2 and CD163; however, total collagen deposition was not a predictor for OS. Conversely, local collagen density, alignment and perpendicular alignment to the tumor boundary (tumor-associated collagen signature-3) were predictors of OS. These results suggest that in invasive carcinoma, the localization of inflammatory cells and aligned collagen orientation predict poor patient survival. Additional clinical studies may help validate whether therapy with selective COX-2 inhibitors alters expression of CD68 and CD163 inflammatory markers.

Objectives:Determine if the extent of parotidectomy or other patient or tumor characteristics influence the rate of sialocele or salivary fistula formation after parotidectomy.Methods:A retrospective chart review was performed for all patients who underwent parotidectomy at the University of Wisconsin from 1994 to 2013. Patients who developed a sialocele or salivary fistula at any time postoperatively were identified. Age, sex, area and size of defect, body mass index (BMI), and rate of malignancy were evaluated to assess any relationship to these complications.Results:A total of 771 patients underwent parotidectomy at our institution from 1994 to 2013. Of these, 75 (9.7%) developed a sialocele or salivary fistula. Sialoceles or salivary fistulas developed in 96% (72/75) within 1 month post‐parotidectomy, and none developed after 6 months. Age, sex, BMI, and histology were not associated with sialocele or salivary fistula formation. Extent of parotidectomy was quantified through assessment of surgical volume of tissue removed. The average volume of tissue removed was 37.8 cubic centimeters (cc) (range 0.2‐277 cc; 95% confidence interval [28.4 cc, 47.2 cc]). Sixty‐four patients (85.3%) underwent superficial parotidectomy, 7 (9.3%) underwent total parotidectomy, and extent of surgery was not documented in 4 patients (5.3%). 2 patients (2.7%) underwent revision surgery. Of patients who underwent superficial parotidectomy, 28 (43.8%) were complete superficial parotidectomy and 23 (35.9%) were partial inferior superficial parotidectomy.Conclusions:Sialocele is an uncommon complication post‐parotidectomy. Contrary to other studies, we observed that sialocele formation does not depend on amount of parotid tissue removed, site of parotidectomy, gender, age, BMI, or rate of malignancy.

Limited data are currently available to direct treatment recommendations in the management of leptomeningeal metastases (LM). Here we review treatment modalities clinicians should understand in order to manage patients with LM. We first describe our institution's experience with the treatment of LM and use this dataset to frame the discussion of LM management. Between 1999 and 2014, 1361 patients with central nervous system metastases were reviewed, 124 (9.1%) had radiographic evidence of LM, and these patients form the cohort for this analysis. Mean age at diagnosis of LM was 52years. Median survival for the entire cohort was 2.3months. The most common primary malignancies were non-small cell lung cancer (25.8%), breast cancer (17.7%), small cell lung cancer (16.9%) and melanoma (8.9%). Univariate analyses demonstrated that greater Karnofsky Performance Status (KPS) (p=0.001) and administration of systemic chemotherapy (p<0.001) resulted in improved median survival. Multivariate Cox analyses revealed that receipt of chemotherapy and a complete course of whole brain radiotherapy (WBRT) (median dose 30Gy in 10 fractions, range 24-40Gy) were predictive of longer survival, (p=0.013 and 0.019, respectively). These data suggest that there is a group of patients with good KPS who may experience significantly longer median survival than expected. Multivariate analysis from this single institution retrospective study demonstrated a benefit for WBRT and chemotherapy in individuals with good KPS. These findings provide contemporary data from a large cohort of LM patients, which may be utilized to guide treatment recommendations, assist in patient counseling and direct future investigations into optimization of treatment regimens.

Objective. To quantify the impact of perioperative beta blocker use on survival after primary cytoreductive surgery for epithelial ovarian cancer.Methods. We conducted a multi-center retrospective study of all women who underwent primary cytoreductive surgery for ovarian cancer (2000 - 2010). One institution had routinely used perioperative blockers for patients "at risk" for coronary events. The other institution did not routinely use perioperative blockers. Demographic, operative, and follow up data were collected. Cox proportional hazards models were used to assess the effect of beta blockers on progression-free interval (PFI) as well as overall survival (OS).Results. Out of 185 eligible patients, 70 received beta blockers and 115 underwent cytoreductive surgery without perioperative blockers. Both groups were similar in demographics. A history of hypertension was present more often in the beta blocker group compared to the group that did not receive beta blockers (22% and 6%, p = 0.002). PFI in beta blocker group was greater at 18.2 vs. 15.8 months (p = 0.66). The OS in the beta blacker group was significantly higher at 44.2 vs. 39.3 months (p = 0.01). In multivariate analysis, perioperative beta locker use was associated with significant improvement in OS (HR 0.68 (0.46-0.99); p = 0.046).Conclusion. Our study showed an association between perioperative beta ocker use and.longer overall survival in patients undergoing primary ovarian cancer cytoreductive surgery. A prospective randomized clinical trial in this population would further validate these results. (C) 2016 Published by Elsevier Inc.

BACKGROUND Angiotensin‐converting enzyme inhibitors (ACEIs) and angiotensin receptor blockers (ARBs) are commonly used antihypertensive medications that have been reported to affect aberrant angiogenesis and the dysregulated inflammatory response. Because of such mechanisms, it was hypothesized that these medications might affect the tumor response to neoadjuvant radiation in patients with rectal cancer. METHODS One hundred fifteen patients who were treated with neoadjuvant radiation at the University of Wisconsin (UW) between 1999 and 2012 were identified. Univariate analyses were performed with anonymized patient data. In a second independent data set, 186 patients with rectal cancer who were treated with neoadjuvant radiation at the Queen's Medical Center of the University of Hawaii (UH) between 1995 and 2010 were identified. These data were independently analyzed as before. Multivariate analyses were performed with aggregate data. RESULTS Among patients taking ACEIs/ARBs in the UW data set, a significant 3‐fold increase in the rate of pathologic complete response (pCR) to neoadjuvant therapy (52% vs 17%, P = .001) was observed. This finding was confirmed in the UH data set, in which a significant 2‐fold–increased pCR rate (24% vs 12%, P = .03) was observed. Identified patient and treatment characteristics were otherwise balanced between patients taking and not taking ACEIs/ARBs. No significant effect was observed on pCR rates with other medications, including statins, metformin, and aspirin. Multivariate analyses of aggregate data identified ACEI/ARB use as a strong predictor of pCR (odds ratio, 4.02; 95% confidence interval, 2.06‐7.82; P < .001). CONCLUSIONS The incidental use of ACEIs/ARBs among patients with rectal cancer is associated with a significantly increased rate of pCR after neoadjuvant treatment. Cancer 2016;122:2487–95 . © 2016 American Cancer Society .

Objective This multi-institutional phase I/II trial explored patient-assessed tolerance of increasingly hypofractionated (HPFX) radiation for low/intermediate risk prostate cancer. Methods 347 patients enrolled from 2002 to 2010. Three increasing dose-per-fraction schedules of 64.7 Gy/22 fx, 58.08 Gy/16 fx and 51.6 Gy/12 fx were each designed to yield equivalent predicted late toxicity. Three quality of life (QoL) surveys were administered prior to treatment and annually upto 3 years. Results Bowel QoL data at 3 years revealed no significant difference among regimens (p = 0.469). Bowel QoL for all regimens declined transiently, largely recovering by three years, with only the 22 fraction decrement reaching significance. Bladder outcomes at 3 years were comparable (p = 0.343) although, for all patients combined, a significant decline was observed from the baseline (p = 0.008). Spitzer quality of life data revealed similarly excellent, 3-year means (p = 0.188). International erectile function data also revealed no significant differences at 3 years although all measures except intercourse satisfaction worsened post-treatment. Conclusions Three-year QoL changes for bowel, bladder and SQLI were modest and similar for 3 HPFX regimens spanning 2.94–4.3 Gy per fraction. These favorable patient-scored outcomes demonstrate the safety and tolerability of such regimens and may be leveraged to support further implementation of mild to moderately hypofractionated radiotherapy in the setting of low and intermediate-risk prostate cancer

Objective: The aim of this study was to determine whether endovascular open revascularization provides an advantageous approach to symptomatic peripheral arterial disease (PAD) over the longer term.Summary of Background Data: The optimal revascularization strategy for symptomatic lower extremity PAD is not established.Methods: We evaluated amputation-Cree survival, overall survival, and relative rate of subsequent vascular intervention after endovascular or open lower extremity revascularization for propensity-score matched cohorts of Medicare beneficiaries with PAD from 2006 through 2009.Results: Among 14,685 eligible patients, 5928 endovascular and 5928 open revascularization patients were included in matched analysis. Patients undergoing endovascular repair had improved amputation-free survival compared with open repair at 30 days (7.4 vs 8.9%, P = 0.002). This benefit persisted over the long term: At 4 years, 49% of endovascular patients had died or received major amputation compared with 54% of open patients (P < 0.001). An endovascular procedure was associated with a risk-adjusted 16% decreased risk of amputation or death compared with open over the study period (hazard ratio: 0.84; 95% confidence interval, 0.79-0.89; P < 0.001). The amputation-free survival benefit associated with an endovascular revascularization was more pronounced in patients with congestive heart failure or ischemic heart disease than in those without (P = 0.021 for interaction term). The rate of subsequent intervention at 30 days was 7.4% greater for the endovascular vs the open revascularization cohort. At 4 years, this difference remained stable at 8.6%.Conclusions: Using population-based data, we demonstrate that an endovascular approach is associated with improved amputation-free survival over the long term with only a modest relative increased risk of subsequent intervention.

Estrogen receptor beta (ER beta) is a potential therapeutic target in triple-negative breast cancer (TNBC). This 2-stage phase 2 study investigated estradiol in advanced TNBC. The study was closed after 17 patients completed the first stage. One patient with an ER beta-positive tumor experienced a prolonged confirmed partial response. Future study of ER beta agonists in selected TNBC may be warranted.Background: Estrogen receptor beta (ER beta) is expressed by 50% to 80% of triple-negative breast cancers (TNBC). Agonism of ER beta has antiproliferative effects in TNBC cells expressing ER beta. This phase 2 study evaluated single-agent high-dose estradiol in patients with advanced TNBC. Patients and Methods: Adult women with measurable advanced TNBC were treated with estradiol 10 mg oral 3 times daily provided continuously for 28-day cycles. A Simon optimal 2-stage design was used. The primary end point was objective response (OR). Secondary end points included progression-free survival (PFS), clinical benefit (CB), and safety. OR, CB, and PFS by ER beta status were also examined. Results: Seventeen evaluable women were enrolled. Median age was 58 years (range, 34-90 years); the median number of prior systemic therapies was 2 (range, 0-6). One patient had a confirmed partial response (OR rate, 5.9%) and remained on the study for > 24 weeks. Three patients had stable disease, with one lasting more than 16 weeks. ER beta expression was detected in 77% (13 patients). The CB rate at 16 weeks was 15% (2 of 13) in ER beta-positive patients and 0% (0 of 4) in ER beta-negative patients (P = 1). PFS was poor (median, 1.9 months) and not statistically significantly different between ER beta-positive versus -negative patients. No new adverse events from estradiol were identified. The study closed after the first stage as a result of limited responses in these unselected patients. Conclusion: In unselected TNBC, high-dose estradiol has limited efficacy. However, further evaluation of ER beta selective agonists in TNBC selected by ER beta expression may be warranted. (C) 2016 Elsevier Inc. All rights reserved.

BACKGROUND:High breast density is linked to an increased risk of breast cancer, and correlates with changes in collagen. In a mouse model of mammary carcinoma in the context of increased collagen deposition, the MMTV-PyMT/Col1a1 (tm1jae) , there is accelerated mammary tumor formation and progression. Previous gene expression analysis suggests that increased collagen density elevates expression of PTGS2 (prostaglandin-endoperoxide synthase 2), the gene for cyclooxygenase-2 (COX-2).METHODS:To understand the role of COX-2 in tumor progression within a collagen-dense microenvironment, we treated MMTV-PyMT or MMTV-PyMT/Col1a1 (tm1jae) tumors prior to and after tumor formation. Animals received treatment with celecoxib, a specific COX-2 inhibitor, or placebo. Mammary tumors were examined for COX-2, inflammatory and stromal cell components, and collagen deposition through immunohistochemical analysis, immunofluorescence, multiplex cytokine ELISA and tissue imaging techniques.RESULTS:PyMT/Col1a1 (tm1jae) tumors were larger, more proliferative, and expressed higher levels of COX-2 and PGE2 than PyMT tumors in wild type (WT) mice. Treatment with celecoxib significantly decreased the induced tumor size and metastasis of the PyMT/Col1a1 tumors, such that their size was not different from the smaller PyMT tumors. Celecoxib had minimal effect on the PyMT tumors. Celecoxib decreased expression levels of COX-2, PGE2, and Ki-67. Several cytokines were over-expressed in PyMT/Col1a1 compared to PyMT, and celecoxib treatment prevented their over-expression. Furthermore, macrophage and neutrophil recruitment were enhanced in PyMT/Col1a1 tumors, and this effect was inhibited by celecoxib. Notably, COX-2 inhibition reduced overall collagen deposition. Finally, when celecoxib was used prior to tumor formation, PyMT/Col1a1 tumors were fewer and smaller than in untreated animals.CONCLUSION:These findings suggest that COX-2 has a direct role in modulating tumor progression in tumors arising within collagen-dense microenvironments, and suggest that COX-2 may be an effective therapeutic target for women with dense breast tissue and early-stage breast cancer.

Abstract Inflammation is an important mediator of tumor progression. The objective of this study was to assess whether the tissue localization of COX-2 and tumor-associated macrophages were associated with clinicopathological features of invasive carcinoma, including collagen deposition and patient survival outcome. A tumor microarray (TMA) of 371 biopsy specimens from patients with invasive breast carcinoma was analyzed for expression of high levels of COX-2, and the macrophage markers CD68 and CD163 in either the tumor nest (TN) or the tumor-associated stroma (TS). The study population for this TMA was female patients, 18 to 80 years of age with a median follow up of 8.4 years. Survival tables were calculated according to the Kaplan-Meier method. We found that elevated collagen deposition was associated with high stromal expression of COX-2 (P < 0.0001); however, the amount of collagen deposition was not a predictor for survival outcome. In order to better analyze patient data, samples were divided into quartiles based on their levels of COX-2 expression and/or macrophage infiltration. Tumor nests with high COX-2 expression had worse patient outcome (P = 0.011). One possible mechanism for this is COX-2 dependent recruitment of macrophages to the tumor microenvironment, supported by our finding of high infiltration of CD68+ macrophages in the TS, and CD163+ macrophages in both TN and TS. This recruitment of macrophages was associated with worse overall survival (OS). Furthermore, patient survival was worsened even more if macrophages expressed COX-2, suggesting positive amplification of COX-2 signaling via macrophage recruitment. This notion is further established by the finding of high presence of CD163+ macrophages in the TN as an independent prognostic factor as revealed by multivariate analysis (P < 0.0001, HR = 4.67). These results suggest that in invasive carcinoma the localization of inflammatory markers within the tumor are biomarkers for patient survival outcome. Therefore, we propose that these patients may benefit from therapy with a selective COX-2 inhibitor such as celecoxib. Citation Format: Karla Esbona, Sandeep Saha, Yanyao Yi, Menggang Yu, Kari Wisinski, Lee G. Wilke, Patricia J. Keely. The presence of COX-2 and tumor-associated macrophages as a prognostic marker for invasive breast carcinoma patients. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3927.

Importance Facial fractures after motor vehicle collisions are a significant source of facial trauma in patients seen at trauma centers. With recent changes in use of seat belts and advances in airbag technology, new patterns in the incidence of facial fractures after motor vehicle collisions have yet to be quantified. Objectives To evaluate the incidence of facial fractures and assess the influence of protective device use in motor vehicle collisions in patients treated at trauma centers in the United States. Design, Setting, and Participants Using a data set from the National Trauma Data Bank, we retrospectively assessed facial fractures in motor vehicle collisions occurring from 2007 through 2012, reported by level I, II, III, and IV trauma centers. Data analysis was performed from March 13 to September 22, 2015. Main Outcomes and Measures We characterized the data set by subsite of facial injury using International Classification of Diseases, Ninth Revision codes including mandible, midface, and nasal fractures. We assessed the influence of variables such as age, sex, race/ethnicity, crash occupant (driver or passenger), use of protective device, and presence or suspicion of alcohol use. Results A total of 518 106 patients required assessment at a trauma center after a motor vehicle collision, with 56 422 (10.9%) experiencing at least 1 facial fracture. Nasal fracture was the most common facial fracture (5.6%), followed by midface (3.8%), other (3.2%), orbital (2.6%), mandible (2.2%), and panfacial fractures (0.8%). Of the subset sustaining at least 1 facial fracture, 5.8% had airbag protection only, 26.9% used a seat belt only, and 9.3% used both protective devices, while 57.6% used no protective device. Compared with no protective device, the use of an airbag alone significantly reduced the likelihood of facial fracture after a motor vehicle collision (odds ratio, 0.82; 95% CI, 0.79-0.86); use of a seat belt alone had a greater effect (odds ratio, 0.57; 95% CI, 0.56-0.58) and use of both devices provided the greatest odds reduction (odds ratio, 0.47; 95% CI, 0.45-0.48). Younger age, male sex, and alcohol use significantly increased the likelihood of facial fracture. Conclusions and Relevance For patients who presented to US trauma centers after motor vehicle collisions between 2007 and 2012, airbags, seat belts, and the combination of the 2 devices incrementally reduced the likelihood of facial fractures. Level of Evidence 3.

Head and neck squamous cell carcinoma (HNSCC) remains a challenging cancer to treat with overall 5-year survival on the order of 50-60%. Therefore, predictive biomarkers for this disease would be valuable to provide more effective and individualized therapeutic approaches for these patients. While prognostic biomarkers such as p16 expression correlate with outcome; to date, no predictive biomarkers have been clinically validated for HNSCC. We generated xenografts in immunocompromised mice from six established HNSCC cell lines and evaluated response to cisplatin, cetuximab, and radiation. Tissue microarrays were constructed from pre- and posttreatment tumor samples derived from each xenograft experiment. Quantitative immunohistochemistry was performed using a semiautomated imaging and analysis platform to determine the relative expression of five potential predictive biomarkers: epidermal growth factor receptor (EGFR), phospho-EGFR, phospho-Akt, phospho-ERK, and excision repair cross-complementation group 1 (ERCC1). Biomarker levels were compared between xenografts that were sensitive versus resistant to a specific therapy utilizing a two-sample t-test with equal standard deviations. Indeed the xenografts displayed heterogeneous responses to each treatment, and we linked a number of baseline biomarker levels to response. This included low ERCC1 being associated with cisplatin sensitivity, low phospho-Akt correlated with cetuximab sensitivity, and high total EGFR was related to radiation resistance. Overall, we developed a systematic approach to identifying predictive biomarkers and demonstrated several connections between biomarker levels and treatment response. Despite these promising initial results, this work requires additional preclinical validation, likely involving the use of patient-derived xenografts, prior to moving into the clinical realm for confirmation among patients with HNSCC.

Author Affiliations: Department of Dermatology, University Clinic of Navarra, Pamplona, Navarra, Spain (Redondo, Gímenez de Azcarate, Aguado); Research Support Service, University Clinic of Navarra, Pamplona, Navarra, Spain (Núñez-Córdoba); Department of Preventive Medicine and Public Health, Medical School, University of Navarra, Pamplona, Navarra, Spain (Núñez-Córdoba); Area of Cell Therapy, University Clinic of Navarra, Pamplona, Navarra, Spain (Andreu, García-Guzman, Prosper).

Introduction: Restoring circulation to patients with symptomatic lower extremity peripheral arterial disease (PAD) is traditionally achieved with open surgery. However, endovascular techniques have enhanced the armamentarium available to patients. No long-term, population-based results exist on which of these provides the optimal approach to symptomatic PAD. Methods: We identified patients who underwent an inpatient endovascular or open lower extremity revascularization for PAD (including those with claudication and limb threat) for 2006 through 2009 from the Centers for Medicare & Medicaid Services Chronic Conditions Warehouse, a 5% national sample of Medicare beneficiaries. The primary outcome was amputation free survival. The secondary outcome was the relative rate of subsequent intervention. Propensity score matching ensured similar baseline characteristics amongst cohorts. Results: Among 14,685 eligible patients, 5,928 endovascular revascularization patients and 5,928 open revascularization patients were included in a matched analysis. Patients undergoing endovascular repair had improved amputation free survival compared to open repair at 30-days (7.4 vs. 8.9%, p=0.002). This benefit persisted over the long-term: At 4-years, 49% of endovascular patients had died or received major amputation compared to 54% of open patients (p<0.001). An endovascular procedure was associated with a risk-adjusted 19% decreased risk of amputation or death compared to open over the study period (hazard ratio: 0.84; 95% confidence interval, 0.79-0.89; p<0.001). The rate of subsequent intervention at 30-days was 7.4% greater for the endovascular versus the open revascularization cohort. At 4-years, this difference remained stable at 8.6%. Conclusions: Using a population-based dataset, we show that an endovascular approach is associated with improved amputation free survival over the long-term. Moreover, we show a modest initial increase in the risk of reintervention after endovascular procedures; the differential rate of reintervention for endovascular patients was less than 10% over four years. The observed benefit associated with endovascular intervention may have significant implications for the millions of patients with PAD.