Citations

Abstract The terminology for human papillomavirus (HPV)–associated squamous lesions of the lower anogenital tract has a long history marked by disparate diagnostic terms derived from multiple specialties. It often does not reflect current knowledge of HPV biology and pathogenesis. A consensus process was convened to recommend terminology unified across lower anogenital sites. The goal was to create a histopathologic nomenclature system that reflects current knowledge of HPV biology, optimally uses available biomarkers, and facilitates clear communication across different medical specialties. The Lower Anogenital Squamous Terminology (LAST) Project was cosponsored by the College of American Pathologists and the American Society for Colposcopy and Cervical Pathology and included 5 working groups; 3 work groups performed comprehensive literature reviews and developed draft recommendations. Another work group provided the historical background and the fifth will continue to foster implementation of the LAST recommendations. After an open comment period, the draft recommendations were presented at a consensus conference attended by LAST work group members, advisors, and representatives from 35 stakeholder organizations including professional societies and government agencies. Recommendations were finalized and voted on at the consensus meeting. The final, approved recommendations standardize biologically relevant histopathologic terminology for HPV-associated squamous intraepithelial lesions and superficially invasive squamous carcinomas across all lower anogenital tract sites and detail the appropriate use of specific biomarkers to clarify histologic interpretations and enhance diagnostic accuracy. A plan for disseminating and monitoring recommendation implementation in the practicing community was also developed. The implemented recommendations will facilitate communication between pathologists and their clinical colleagues and improve accuracy of histologic diagnosis with the ultimate goal of providing optimal patient care.

Abstract Background: Alveolar rhabdomyosarcoma (ARMS) is an aggressive pediatric cancer showing skeletal muscle differentiation. New therapeutic targets are required to improve the dismal cure rates for patients with metastatic ARMS. Protein Kinase C iota (PRKCI) has been shown to play an important role in many cancers but little to date is known of this kinase's role in rhabdomyosarcoma. Materials and Methods: Human samples were obtained from the Pediatric Cooperative Human Tissue Network (Columbus, OH). Mouse samples were obtained from a Myf6Cre, Pax3: Fkhr, p53 conditional mouse model of ARMS and a Pax7CreER, Ptch1, p53 conditional model of embryonal rhabdomyosarcoma (ERMS). Expression of PKCι in tumors was evaluated by quantitative RT-PCR, western blotting and immunohistochemistry. To elucidate the functional role of PKCι in ARMS, we performed invasion assays, anchorage-independent colony formation assays and in vitro growth inhibition assays using primary cell lines obtained from mouse ARMS tumors, treating either with the PKCι specific inhibitor Aurothiomalate (ATM) or by PKCι RNA interference. Results: Over-expression of PKCι at the mRNA and protein level was confirmed in human and mouse ARMS and ERMS. Immunoblotting confirmed that PKCι and its downstream targets, mitogen-activated protein kinase and nuclear factor-kappa B were expressed and highly activated in primary cell cultures. Inhibition of PKCι by RNA interference resulted in a dramatic decrease in cell viability and colony formation. Treatment of primary cell culture using ATM resulted in decreased cell viability (IC50: 54.3 – 256.9 uM) and colony formation, and increased G2/M blockade. Co-treatment with ATM and vincristine, a microtubule inhibitor currently used in rhabdomyosarcoma treatment regimens, resulted in a Combination Index of 0.572 – 1.017, suggesting cooperativity between these agents. Conclusion: These results suggest that PKCι has an important function in proliferation of ARMS and that the PKCι specific inhibitor ATM shows activity in ARMS at clinically-feasible drug concentrations. Furthermore, these results suggest that combination therapy with vincristine and ATM should be investigated further in ARMS. Ken Kikuchi and Anuradha Soundararajan contributed this presentation equally. 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 1043. doi:10.1158/1538-7445.AM2011-1043

Abstract Motivation: KRAS is commonly mutated in a variety of cancers including lung cancer. The KRAS gene is frequently mutated at codons 12 and 13 in lung adenocarcinomas in patients with a history of smoking. Tumors harboring an activating KRAS mutation are aggressive and are often resistant to available therapies. In the present study, we set out to identify pathways that are specifically altered in lung cancer patients whose tumors harbor a KRAS mutation using next generation sequencing technology. Methods: We performed 50nt paired-end RNA-sequencing in 15 lung adenocarcinoma samples (8 and 7 samples with and without KRAS mutation). Reads were aligned to genome and exon junctions using Illumina's alignment tool ELAND_RNA. CASAVA and Genome Studio data analysis software were used to obtain read counts for genes, exons and exon junctions. We applied a variety of computational methods (Casper R package, Bowtie, TopHat) and softwares (Partek, R statistical software, JMP, Ingenuity Pathway Anlaysis, DAVID) to carry out our analyses. Results: Differential gene expression and splicing analysis were performed between the two groups. We identified 115 genes that were differentially expressed and 112 genes that consist of splicing variants with at least 2 fold changes and p-value < 0.01. We randomly selected 6/15 samples and performed real-time qRT-PCR for 6 differentially expressed genes. Significant correlations were observed ranging from 0.44 to 0.83, when qRT-PCR results were compared with RNA-sequencing expression data. Pathway analysis with 115 differentially expressed genes and 112 splicing variants revealed that the most significant pathway is composed entirely of NFκB focus genes, suggesting that there is a direct connection between oncogenic KRAS and activation of the NFκB signaling pathway. Conclusions: NFκB has been implicated in KRas-mediated formation of early stage lung adenomas in the LSL-Kas mouse model. Our data indicate that this connection also occurs in human lung cancer and establish NFκB activation as a key manifestation of KRAS mutation in human lung cancer. Hence drugs targeting NFκB and NFκB related genes may potentially be helpful for the treatment of patients with oncogenic KRAS mutations. (Supported in part by grants from the 26.2 with Donna Foundation and the Florida Department of Health Bankhead/Coley program.) 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 4975. doi:10.1158/1538-7445.AM2011-4975

Abstract Poly (ADP-ribose) polymerase (PARP) inhibition can enhance the efficacy of temozolomide (TMZ) and has been shown to significantly prolong survival in orthotopic GBM xenografts. Here the effects of PARP inhibitor AG014699 (AG699) on the efficacy of TMZ was evaluated and corresponding pharmacokinetic and pharmacodynamic studies were performed in the MGMT methylated GBM12 primary GBM xenograft line. AG699 effectively sensitized GBM12 cells to TMZ in vitro in a neurosphere formation assay: IC50 of 1.56 M for TMZ when combined with 1 μM AG699 compared to 8.16 M TMZ when used alone). Using these same cells, orthotopic xenografts were established and mice were randomized to 3–28 day cycles of placebo or TMZ (50 mg/kg days 1–5) with or without AG699 (1 mg/kg IP days 1–5). Although this model is responsive to other TMZ/PARP inhibitor combinations, the addition of AG699 was ineffective (median survival: placebo 16 days, TMZ 68 days and TMZ/AG699 81 days, p=0.883). In contrast, AG699 significantly potentiated TMZ-induced tumor regrowth delay in a flank tumor model with GBM12 (median time for tumors to reach endpoint: placebo 32 days, TMZ 86 days and TMZ/AG699 121 days, p<0.01). To more clearly understand the discrepancy between intracranial and flank studies, drug accumulation and target inhibition were assessed. Plasma and tissue distribution studies in mice bearing GBM12 orthotopic xenografts showed that 0.5 h after AG699 treatment its level in brain and tumor xenograft was much lower compared with that in liver (see table). In an assessment of catalytic activity, PARP activity in liver was inhibited more than 95% of control at least 2 h after AG699 treatment, while it was inhibited less than 50% and 30% in orthotopic xenograft tissue and normal brain, respectively (Table). Collectively, these data support the hypothesis that the efficacy of AG699 combined with TMZ is limited in the orthotopic model due to limited drug accumulation within the brain, and the mechanisms of which are under investigation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr A77.

Oncolytic reovirus is carried by cells to tumors and protected from neutralizing antibodies in the circulation.

Background —A multinational, randomized, placebo-controlled trial (Platelet Glycoprotein IIb/IIIa in Unstable Angina: Receptor Suppression Using Integrilin Therapy, PURSUIT) demonstrated that the platelet glycoprotein IIb/IIIa receptor antagonist eptifibatide reduced the incidence of death or myocardial infarction among patients with acute ischemic syndromes without ST-segment elevation. Because of expected differences in practice patterns, a prospectively planned analysis of outcomes as a function of regions of the world was performed. The current study provides a detailed assessment of eptifibatide among the subgroup of patients enrolled within the United States. Methods and Results —Patients presenting with chest pain within the previous 24 hours and ischemic ECG changes or creatine kinase–MB elevation were eligible for enrollment. Of the 10 948 patients randomized worldwide, 4035 were enrolled within the United States. Patients were allocated to placebo or eptifibatide infusion for up to 72 to 96 hours. Other medical therapies and revascularization strategies were at the discretion of the treating physician. Eptifibatide reduced the rate of the primary end point of death or myocardial infarction by 30 days from 15.4% to 11.9% ( P =0.003) among patients in the United States. The treatment effect was achieved early and maintained over a period of 6 months (18.9% versus 15.2%; P =0.004). Bleeding events were more common in patients receiving eptifibatide but were predominantly associated with invasive procedures. The magnitude of clinical benefit from eptifibatide was greater among patients in the United States than elsewhere in the world. Conclusions —Platelet glycoprotein IIb/IIIa receptor blockade with eptifibatide reduces the incidence of death or myocardial infarction among patients treated for acute ischemic syndromes without ST-segment elevation within the United States.

BackgroundRight heart failure is an important cause of morbidity and mortality in primary pulmonary hypertension. In a recent prospective, randomized study of severely symptomatic patients, treatment with prostacyclin (epoprostenol) produced improvements in hemodynamics, quality of life, and survival. This article describes the echocardiographic characteristics of participants in this trial; the relationships of echocardiographic variables to hemodynamic parameters, exercise capacity, and quality of life; and the echocardiographic changes associated with prostacyclin therapy.Methods and ResultsThe 81 patients enrolled in this multicenter trial were randomized to treatment with a long-term infusion of prostacyclin in addition to conventional therapy (n=41) or conventional therapy alone (n=40) for 12 weeks. Echocardiograms and assessments of hemodynamics, exercise capacity, and quality of life were performed before and after the treatment phase. On baseline evaluation, patients had marked right ventricular dilatation and dysfunction, abnormal septal curvature, and significant tricuspid regurgitation with a high regurgitant velocity. Pericardial effusions were common. More pronounced abnormalities in right heart structure and function were associated with higher pulmonary arterial and mean right atrial pressures, lower cardiac index, and impaired exercise capacity but had no predictable relationship to quality-of-life indicators. The 12-week infusion of prostacyclin had beneficial effects on right ventricular size, curvature of the interventricular septum, and maximal tricuspid regurgitant jet velocity.ConclusionsThe echocardiographic manifestations of severe primary pulmonary hypertension reflect abnormalities in hemodynamics and exercise capacity. Prostacyclin has beneficial effects on right heart structure and function that may contribute to the clinical improvement and prolonged survival observed with this drug.

A study has been made at the Mayo Clinic of the results of treatment in 28 hypertensive patients receiving thiazide diuretics. The use of these agents produced a significant but very temporary reduction in plasma volume in most patients. The plasma volume reverted to or toward normal after the first week of treatment; therefore, hypovolemia could not explain the prolonged hypotensive effect of these drugs. Administration of thiazide diuretics did not deplete the body of its sodium stores, as measured by isotope studies in five patients on an unrestricted diet. Therapy with thiazides did not produce a persistent increase in urinary aldosterone excretion in the nine patients studied for this possibility.

Objectives Chronic constipation in diabetes mellitus is associated with colonic motor dysfunction and is managed with laxatives. Cholinesterase inhibitors increase colonic motility. This study evaluated the effects of a cholinesterase inhibitor on gastrointestinal and colonic transit and bowel function in diabetic patients with constipation. Design After a 9-day baseline period, 30 patients (mean±SEM age 50±2 years) with diabetes mellitus (18 type 1, 12 type 2) and chronic constipation without defaecatory disorder were randomised to oral placebo or pyridostigmine, starting with 60 mg three times a day, increasing by 60 mg every third day up to the maximum tolerated dose or 120 mg three times a day; this dose was maintained for 7 days. Gastrointestinal and colonic transit (assessed by scintigraphy) and bowel function were evaluated at baseline and the final 3 and 7 days of treatment, respectively. Treatment effects were compared using analysis of covariance, with gender, body mass index and baseline colonic transit as covariates. Results 19 patients (63%) had moderate or severe autonomic dysfunction; 16 (53%) had diabetic retinopathy. 14 of 16 patients randomised to pyridostigmine tolerated 360 mg daily; two patients took 180 mg daily. Compared with placebo (mean±SEM 1.98±0.17 (baseline), 1.84±0.16 (treatment)), pyridostigmine accelerated (1.96±0.18 (baseline), 2.45±0.2 units (treatment), p<0.01) overall colonic transit at 24 h, but not gastric emptying or small-intestinal transit. Treatment effects on stool frequency, consistency and ease of passage were significant (p≤0.04). Cholinergic side effects were somewhat more common with pyridostigmine (p=0.14) than with placebo. Conclusions Cholinesterase inhibition with oral pyridostigmine accelerates colonic transit and improves bowel function in diabetic patients with chronic constipation. Clinical trial registration number TrialRegNo (NCT 00276406).