Keeney, Gary L.

Aneurysmal bone cysts (ABC) are rare, benign, expansile lesions of bone often found in the metaphyses of long bones in pediatric and young adult population. Multiple fluid levels are typically seen on imaging with magnetic resonance imaging (MRI) or computed tomography (CT). We describe a case of a primary ABC in the fibula of a 34-year-old man diagnosed on ultrasound with a mobile fluid level demonstrated sonographically.

Abstract Background: Ovarian carcinoma is composed of five major histologic types, which associate with outcome and predict therapeutic response. Our aim was to evaluate histologic type assessments across the centers participating in the Ovarian Tumor Tissue Analysis (OTTA) consortium using an immunohistochemical (IHC) prediction model. Methods: Tissue microarrays (TMA) and clinical data were available for 524 pathologically confirmed ovarian carcinomas. Centralized IHC was conducted for ARID1A, CDKN2A, DKK1, HNF1B, MDM2, PGR, TP53, TFF3, VIM, and WT1, and three histologic type assessments were compared: the original pathologic type, an IHC-based calculated type (termed TB_COSPv2), and a WT1-assisted TMA core review. Results: The concordance between TB_COSPv2 type and original type was 73%. Applying WT1-assisted core review, the remaining 27% discordant cases subdivided into unclassifiable (6%), TB_COSPv2 error (6%), and original type error (15%). The largest discordant subgroup was classified as endometrioid carcinoma by original type and as high-grade serous carcinoma (HGSC) by TB_COSPv2. When TB_COSPv2 classification was used, the difference in overall survival of endometrioid carcinoma compared with HGSC became significant [RR 0.60; 95% confidence interval (CI), 0.37–0.93; P = 0.021], consistent with previous reports. In addition, 71 cases with unclear original type could be histologically classified by TB_COSPv2. Conclusions: Research cohorts, particularly those across different centers within consortia, show significant variability in original histologic type diagnosis. Our IHC-based reclassification produced more homogeneous types with respect to outcome than original type. Impact: Biomarker-based classification of ovarian carcinomas is feasible, improves comparability of results across research studies, and can reclassify cases which lack reliable original pathology. Cancer Epidemiol Biomarkers Prev; 22(10); 1677–86. ©2013 AACR.

Abstract Background. The hepatocyte growth factor (HGF) signaling pathway regulates key cellular functions related to tumor invasion and metastasis including cell motility, cell proliferation, and morphogenesis. We previously found that minor alleles at a single-nucleotide polymorphism (SNP; rs5745709) in the gene encoding the cytokine HGF was associated with poorer overall survival in women with invasive epithelial ovarian cancer (EOC) enrolled at Mayo Clinic (MAY) and The Cancer Genome Atlas (TCGA) (HR 1.6, 95% CI 1.3-1.9, p=8.9×10−5). In addition, we found that that genotype correlated with HGF expression in ovarian cancer tissue from TCGA women, suggesting that genetically-regulated expression of HGF may play a role in post-diagnosis survival. To further examine this possibility, we evaluated intensity of antibody immunostaining for HGF, MET (HGF's transmembrane tyrosine kinase receptor), and phospho-MET (pMET; MET's activated form) in tumor cells using tissue microarrays (TMAs) created from genotyped primary peritoneal and EOC cases, and correlated these expression levels with overall survival and HGF SNP genotypes. Methods. Participants were 326 white non-Hispanic women over 20 years of age with histologically-confirmed invasive EOC diagnosed between 1998 and 2009. All cases were enrolled at MAY within one year of diagnosis. Five μm sections of TMAs including three 0.6 mm cores per case were immunostained with HGF, MET, and pMET primary antibodies. Staining levels were classified according to combinations of staining extent and intensity as negative (< 10% cells stained), weak, moderate, or strong. The maximal expression value recorded over the multiple cores for each case was used. Cox proportional hazards regression was used to examine associations of overall survival with HGF, MET and pMET expression levels, adjusted for relevant clinical variables. Associations between expression levels and HGF SNP rs5745709 genotypes were examined using linear regression models. Results. HGF and MET expression were not associated with overall survival in our series of EOC cases (p=0.48 and 0.90, respectively). pMET expression was associated with survival, but in the opposite direction of that hypothesized (HR 0.77, 95% CI 0.39-1.52 for moderate expression; HR 0.55, 95% CI 0.27-1.10 for strong expression; each referent to negative/weak, p=0.01). Expression levels were not significantly associated with HGF rs5745709 genotypes (p=0.80 for HGF expression, p=0.89 for MET, p=0.41 for pMET). Conclusion. HGF expression levels were not associated with overall survival or with HGF SNP genotypes that correlated with survival in a previous study. Functional studies such as this one are crucial as a follow-up to discovery-based studies in order to assess biologic plausibility of observed results. 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 893. doi:10.1158/1538-7445.AM2011-893

Abstract Background: Invasive ovarian cancer is a significant cause of gynecologic cancer mortality. Methods: We examined whether this mortality was associated with inherited variation in approximately 170 candidate genes/regions [993 single-nucleotide polymorphisms (SNPs)] in a multistage analysis based initially on 312 Mayo Clinic cases (172 deaths). Additional analyses used The Cancer Genome Atlas (TCGA; 127 cases, 62 deaths). For the most compelling gene, we immunostained Mayo Clinic tissue microarrays (TMA, 326 cases) and conducted consortium-based SNP replication analysis (2,560 cases, 1,046 deaths). Results: The strongest initial mortality association was in HGF (hepatocyte growth factor) at rs1800793 (HR = 1.7, 95% CI = 1.3–2.2, P = 2.0 × 10−5) and with overall variation in HGF (gene-level test, P = 3.7 × 10−4). Analysis of TCGA data revealed consistent associations [e.g., rs5745709 (r2 = 0.96 with rs1800793): TCGA HR = 2.4, CI = 1.4–4.1, P = 2.2 × 10−3; Mayo Clinic + TCGA HR = 1.6, CI = 1.3–1.9, P = 7.0 × 10−5] and suggested genotype correlation with reduced HGF mRNA levels (P = 0.01). In Mayo Clinic TMAs, protein levels of HGF, its receptor MET (C-MET), and phospho-MET were not associated with genotype and did not serve as an intermediate phenotype; however, phospho-MET was associated with reduced mortality (P = 0.01) likely due to higher expression in early-stage disease. In eight additional ovarian cancer case series, HGF rs5745709 was not associated with mortality (HR = 1.0, CI = 0.9–1.1, P = 0.87). Conclusions: We conclude that although HGF signaling is critical to migration, invasion, and apoptosis, it is unlikely that HGF genetic variation plays a major role in ovarian cancer mortality. Furthermore, any minor role is not related to genetically-determined expression. Impact: Our study shows the utility of multiple data types and multiple data sets in observational studies. Cancer Epidemiol Biomarkers Prev; 20(8); 1638–48. ©2011 AACR.

Abstract Using data from patients with invasive epithelial ovarian cancer, we sought to examine whether overall survival was associated with common inherited variation in candidate genes and genomic regions including those involved in angiogenesis, mitosis, and regions of interest from genome-wide association and expression studies. We conducted a two-stage analysis based on Mayo Clinic patients with replication of peak results in the Cancer Genome Atlas (TCGA). Ovarian cancer patients (N=312) seen at the Mayo Clinic diagnosed from 1999 to 2006 with median follow-up of 3.7 years (range, 0.1 − 9.5 years) and 172 observed deaths were analyzed at 993 single nucleotide polymorphisms (SNPs) in 179 genes. For all patients and for serous patients only (N=192 with 129 observed deaths), Cox regression was used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for each SNP, adjusted for other prognostic factors. Twenty-nine SNPs in ten genes revealed p < 0.01 among all cases or among serous cases only, including 21 independent SNP at r2=0.9; analyses within four genes (HGF, PLG, PRKACB, and DCTN5) yielded p < 0.001. The strongest result was in HGF at rs1800793 (all cases: HR 1.7, 95% CI 1.3 − 2.2, p-trend 1.9 E-5; serous cases: HR 1.6, 95% CI 1.2 − 2.2, p-trend 1.8 E-3). A correlated SNP rs2214825 also showed association (r2=0.76; all cases: HR 1.4, 95% CI 1.1 − 1.8, p-trend 3.3 E-3; serous cases: HR 1.4, 95% CI 1.0 − 1.9, p-trend 0.03), and PCA gene-level tests of the HGF locus were also significantly associated with survival (all cases, p=3.7 E-4; serous cases, p=0.01). Among all cases, differential survival was also associated with DCTN5 rs12447304 (HR 1.8, 95% CI 1.3-2.6; p=7.1 E-4), and among serous cases, with PLG rs783173 (HR 1.6, 95% CI 1.2 − 2.0, p = 8.1 E-4) and PRKACB rs1402694 (HR 1.7, 95% CI 1.3 − 2.2, p=3.2 E-4). We then examined SNPs in the most suggestive gene, HGF, using data from TCGA which had genotyped four overlapping HGF SNPs in 350 invasive cases (188 deaths). Covariate-adjusted TCGA analysis at rs2214825 revealed consistent results (HR 1.7, 95% CI 1.3-2.2, p=1.4 E-4) as did analyses of three other HGF SNPs (HR range 1.3 to 1.5, p range 3.4 E-3 to 0.049); correlation between SNP genotypes and HGF expression was also observed (p=1.4 E-3, rs2214825, probe 210997_at). While this is the first report of SNPs in HGF, it has long been known that the HGF signaling pathway, activated by HGF ligation of c-MET, plays a key role ovarian cancer cell growth, migration, and invasion. c-MET is overexpressed in 40%-60% of ovarian tumors, and high overexpression is associated with lower overall survival. In addition, HGF has been shown to enhance the response of ovarian cancer cells to low doses of paclitaxel and cisplatin by activating the intrinsic apoptotic pathway, leading to cancer cell death. These results now suggest that inherited differences in HGF among women with invasive disease may play a role in survival, holding potential clues for tailored avenues of treatment. 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 897.

Abstract Edmonston vaccine strains of measles virus (MV) have shown significant antitumor activity in preclinical models of ovarian cancer. We engineered MV to express the marker peptide carcinoembryonic antigen (MV-CEA virus) to also permit real-time monitoring of viral gene expression in tumors in the clinical setting. Patients with Taxol and platinum-refractory recurrent ovarian cancer and normal CEA levels were eligible for this phase I trial. Twenty-one patients were treated with MV-CEA i.p. every 4 weeks for up to 6 cycles at seven different dose levels (103–109 TCID50). We observed no dose-limiting toxicity, treatment-induced immunosuppression, development of anti-CEA antibodies, increase in anti-MV antibody titers, or virus shedding in urine or saliva. Dose-dependent CEA elevation in peritoneal fluid and serum was observed. Immunohistochemical analysis of patient tumor specimens revealed overexpression of measles receptor CD46 in 13 of 15 patients. Best objective response was dose-dependent disease stabilization in 14 of 21 patients with a median duration of 92.5 days (range, 54–277 days). Five patients had significant decreases in CA-125 levels. Median survival of patients on study was 12.15 months (range, 1.3–38.4 months), comparing favorably to an expected median survival of 6 months in this patient population. Our findings indicate that i.p. administration of MV-CEA is well tolerated and results in dose-dependent biological activity in a cohort of heavily pretreated recurrent ovarian cancer patients. Cancer Res; 70(3); 875–82

AbstractFolate receptor α (FRα) expression in epithelial ovarian cancer may be related to folate intake. We examined this association using multivitamin intake, a proxy for folic acid, and assessed whether the relation was modified by alcohol intake, a folate agonist. Cases (n = 148) with suspected epithelial ovarian cancer, of ages ≥20 years, were seen at Mayo Clinic, Minnesota, between 2000 and 2004; those with tumor specimens (n = 108) were included in analyses. Outpatient controls (n = 148) without cancer and with at least one ovary intact were matched to cases by age (within 5 years) and state of residence. Multivitamin (≥4 pills/wk) and weekly alcohol (≥5 drinks) intakes were assessed. Tumor specimens were analyzed immunohistochemically for FRα. Multivariable rate ratios (RR) and 95% confidence intervals (CI) were estimated using unconditional logistic regression. In case-control analysis, the RRs of multivitamin intake with absent/weak/moderate and strong-expressing FRα tumors were 0.30 (95% CI, 0.12-0.70) and 0.47 (95% CI, 0.24-0.91), respectively. For alcohol, the associations were 0.84 (95% CI, 0.24-2.86) and 1.65 (95% CI, 0.69-3.93), respectively. In case-case analysis, the RR associated with developing strong-expressing versus other FRα tumors was 3.13 (95% CI, 1.14-8.65) for multivitamins and 1.58 (95% CI, 0.45-5.60) for alcohol. The data did not support evidence for an interaction between multivitamin and alcohol intake with risk of developing a strong-expressing FRα tumor. The association of multivitamin intake with ovarian cancer may depend on FRα expression level.