Biomarker Development in Endometrial Cancer: Circulating Tumour Cells, Tissue Methylation and Genotyping Studies

Endometrial cancer (EC) is the most common gynaecological malignancy in the developed world and 4th most common women’s cancer in the UK. Although approximately 75% of women present with surgically resectable disease, 20% of these will relapse and 25% of initial diagnoses occur with metastatic disease. There are currently no validated biomarkers to assess treatment response or target molecular therapies, despite evidence for targetable aberrations in the literature. The PI3K pathway in particular is commonly mutated in EC and stathmin is a recently identified phosphoprotein associated with PI3K pathway activation and with prognostic significance in EC. I investigated biomarker development and novel pathways in EC in two ways: firstly, through a feasibility study of circulating tumour cell (CTC) enumeration and molecular profiling (MP) in patients with advanced endometrioid and non-endometrioid EC (EEC and NEEC); and secondly, through methylation and copy number variation (CNV) studies on formalin-fixed paraffin-embedded (FFPE) and fresh frozen (FF) EEC. CTCs have prognostic and predictive significance in a number of cancers, with increasing evidence on CTC MP. CTC enumeration and assessment of stathmin overexpression was performed on the validated Veridex CellSearch platform and shown to be feasible. In addition, CTC positivity was associated with worse survival and there was a subset of patients for whom a positive CTC count was predictive of outcome on chemotherapy. The second component focused on methylation and CNV analysis in the different phases of endometrial carcinogenesis from normal endometrium to atypical endometrial hyperplasia (AEH) and EEC. By extracting DNA from FFPE and FF EEC and using the Illumina Infinium HumanMethylation450 BeadChip, I was able to demonstrate these methods were feasible and that differential methylation and CNV was evident in the transition from normal endometrium through to AEH and EEC. This research provides a basis for further biomarker development and novel target selection in EC

Distal ileal stenosing subserosal lipoma: A case report

Lipoma is the commonest soft tissue tumour and ubiquitous in distribution.The gastrointestinal tract is a rare site for this neoplasm.This 38 years old patient presented to the surgical emergency unit of the Jos University Teaching Hospital with features of intestinal obstruction which was confirmed by plain abdominal X-ray. Patient was resuscitated and had exploratory laparotomy. At surgery,a dilated,oedematous,and pale segment of ileum was seen measuring 56cm in length and 10cm short of the ileo-caecal junction, where an obstruction had occurred.The distal segment was collapsed.A limited right hemi-colectomy was done with ileocolic anastomosis.Specimen received at the Histopathology Laboratory consisted of 45cm of the ileum,the caecum,appendix,and proximal 25cm of the colon in continuity.There was stenosis affecting the distal 30cm of the ileum.The wall of the stenosed part of ileum had intramural fat at the sub-serosal locale. Histology confirmed the presence of sheets of matured adipocytes between the muscularispropria and serosa.Patient condition improved and was discharged seven days after surgery.This case is reported five months after surgery.We recommend that lipoma be at all times considered in the differential diagnosis of intestinal obstruction.Keywords: Subserosal lipoma,ileal stenosis,intestinal obstructio

Value of the visual prostate symptom score in evaluation of symptomatic benign prostatic enlargement: prospective study in a Nigerian population

Background: To evaluate the correlation of Visual Prostate Symptom Score (VPSS) with International Prostate Symptom Score (IPSS) and Maximum Urinary Flow (Qmax). To investigate the effect of educational level on the ability to independently complete the VPSS versus the IPSS and time taken to do so.Methods: Bio data was taken from men with lower urinary tract symptoms (LUTS) due to Benign Prostatic Enlargement (BPE) who presented at the Urology clinic of Jos University Teaching Hospital. They were administered the IPSS questionnaire and VPSS pictogram, which they completed with or without physician assistance and the time taken to do so was noted. They subsequently had uroflowmetry done on same visit and the data was recorded in a structured proforma. Statistical analysis was done using SPSS(R) version 20. Correlation test was done for VPSS, IPSS and Qmax while the paired t-test was used for the average time spent in completing both questionnaires. A p-value <0.05 was considered as significant.Results: Eighty-five men (aged 42 to 94 years) were enrolled in the study. The VPSS correlated significantly with the IPSS in terms of total score (r = +0.684, p<0.001) and QoL (r = +0.570, p<0.001), as well as with the Qmax (r = -0.222, p = 0.041). A greater proportion (21.2%) of men with limited education could complete the VPSS without physician assistance as compared to the IPSS (6.0%) and the average time taken to complete the VPSS (170.51 seconds) was significantly shorter than the time taken to complete the IPSS (406.42 seconds).Conclusions: The VPSS correlates significantly with the IPSS and Qmax. It can be completed without physician assistance by a greater proportion of men with limited education within a shorter time period

Enumeration and Molecular Characterisation of Circulating Tumour Cells in Endometrial Cancer

BACKGROUND: This is a feasibility study to determine whether circulating tumour cells (CTCs) are detectable and suitable for molecular profiling in advanced endometrial cancer (aEC). METHOD: Between October 2012 and February 2014, 30 patients with aEC had baseline and up to 3 follow-up samples. CTCs and stathmin expression were evaluated using the CellSearch platform. Epithelial cell adhesion molecule (EpCAM) and stathmin immunohistochemistry were performed on FFPE tumour tissue. RESULTS: Eighteen from 30 (60%) patients had detectable CTCs during study [1 CTC (n = 7), 2 (n = 4), 3 (n = 1), 4 (n = 2), 7 (n = 1), 8 (n = 1), 22 (n = 1), 172 (n = 1) in 7.5 ml blood]. Ten from 18 patients had between 50 and 100% of detectable CTCs that were stathmin positive. More CTC-positive than CTC-negative patients had non-endometrioid versus endometrioid histology, tumour size ≥5 versus 0.05, 95% confidence interval 0.7-16.2]. Twenty-one tumour blocks were tested for EpCAM and stathmin immunohistochemistry (IHC). Stathmin tumour immunostaining scores (TIS) on IHC were higher in CTC-positive patients. CONCLUSION: CTC enumeration and molecular profiling with stathmin on the CellSearch platform is feasible in aEC. Stathmin TIS on IHC, a known prognostic marker in EC, was associated with CTC positivity

Phase 1 study of cardiac safety of TAS-102 in patients with advanced solid tumors

PURPOSE: TAS-102 is a novel oral agent combining the antineoplastic thymidine-based nucleoside analogue, trifluridine, and the thymidine phosphorylase inhibitor, tipiracil (molar ratio 1:0.5). TAS-102 has shown good activity in refractory metastatic colorectal cancer with acceptable safety. No QT prolongation was seen in clinical studies. This study aimed to investigate TAS-102 cardiac safety for regulatory requirements. METHODS: This was a phase 1, non-randomized study in adults with advanced solid tumors. Intensive QT assessments were conducted at baseline, placebo, and following single and multiple doses of TAS-102 during a 28-day cycle. RESULTS: Following single- and multiple-dose administration (N = 30), the upper bounds of the one-sided 95 % confidence intervals for the difference between TAS-102 and placebo in time-matched baseline-subtracted 12-lead Holter QT intervals did not exceed 20 ms at any prespecified time point. One patient had a change from baseline in QTcI interval ≥60 ms, and one patient had a QTcI interval >500 ms following multiple-dose TAS-102 administration. No patient had an uncorrected QT, QTcF, or QTcB interval >500 ms. Based on the exposure-response analysis between TAS-102 plasma concentrations and the placebo-adjusted QTc intervals, none of the upper bounds of the one-sided 95 % prediction intervals exceeded 20 ms. There were no significant morphological changes for T or U waves. No cardiovascular AEs were reported in cycle 1. Across all cycles, no patient experienced an AE of ventricular tachycardia, ventricular fibrillation, syncope, or seizure. CONCLUSIONS: There was no clinically relevant relationship between TAS-102 plasma concentrations and QTc interval; TAS-102 had no clinically relevant effects on cardiac repolarization

Phase 1/2 study of daratumumab, lenalidomide, and dexamethasone for relapsed multiple myeloma

Daratumumab, a human CD38 immunoglobulin G1 kappa (IgG1κ) monoclonal antibody, has activity as monotherapy in multiple myeloma (MM). This phase 1/2 study investigated daratumumab plus lenalidomide/dexamethasone in refractory and relapsed/refractory MM. Part 1 (dose escalation) evaluated 4 daratumumab doses plus lenalidomide (25 mg/day orally on days 1-21 of each cycle) and dexamethasone (40 mg/week). Part 2 (dose expansion) evaluated daratumumab at the recommended phase 2 dose (RP2D) plus lenalidomide/dexamethasone. Safety, efficacy, pharmacokinetics, immunogenicity, and accelerated daratumumab infusions were studied. In part 1 (13 patients), no dose-limiting toxicities were observed, and 16 mg/kg was selected as the R2PD. In part 2 (32 patients), median time since diagnosis was 3.2 years, with a median of 2 prior therapies (range, 1-3 prior therapies), including proteasome inhibitors (91%), alkylating agents (91%), autologous stem cell transplantation (78%), thalidomide (44%), and lenalidomide (34%); 22% of patients were refractory to the last line of therapy. Grade 3 to 4 adverse events (≥5%) included neutropenia, thrombocytopenia, and anemia. In part 2, infusion-related reactions (IRRs) occurred in 18 patients (56%); most were grade ≤2 (grade 3, 6.3%). IRRs predominantly occurred during first infusions and were more common during accelerated infusions. In part 2 (median follow-up of 15.6 months), overall response rate was 81%, with 8 stringent complete responses (25%), 3 complete responses (9%), and 9 very good partial responses (28%). Eighteen-month progression-free and overall survival rates were 72% (95% confidence interval, 51.7-85.0) and 90% (95% confidence interval, 73.1-96.8), respectively. Daratumumab plus lenalidomide/dexamethasone resulted in rapid, deep, durable responses. The combination was well tolerated and consistent with the safety profiles observed with lenalidomide/dexamethasone or daratumumab monotherapy. This trial was registered at www.clinicaltrials.gov as #NCT01615029

Role of DNA methylation and epigenetic silencing of HAND2 in endometrial cancer development

Background: Endometrial cancer incidence is continuing to rise in the wake of the current ageing and obesity epidemics. Much of the risk for endometrial cancer development is influenced by the environment and lifestyle. Accumulating evidence suggests that the epigenome serves as the interface between the genome and the environment and that hypermethylation of stem cell polycomb group target genes is an epigenetic hallmark of cancer. The objective of this study was to determine the functional role of epigenetic factors in endometrial cancer development. Methods and Findings: Epigenome-wide methylation analysis of >27,000 CpG sites in endometrial cancer tissue samples (n = 64) and control samples (n = 23) revealed that HAND2 (a gene encoding a transcription factor expressed in the endometrial stroma) is one of the most commonly hypermethylated and silenced genes in endometrial cancer. A novel integrative epigenome-transcriptome-interactome analysis further revealed that HAND2 is the hub of the most highly ranked differential methylation hotspot in endometrial cancer. These findings were validated using candidate gene methylation analysis in multiple clinical sample sets of tissue samples from a total of 272 additional women. Increased HAND2 methylation was a feature of premalignant endometrial lesions and was seen to parallel a decrease in RNA and protein levels. Furthermore, women with high endometrial HAND2 methylation in their premalignant lesions were less likely to respond to progesterone treatment. HAND2 methylation analysis of endometrial secretions collected using high vaginal swabs taken from women with postmenopausal bleeding specifically identified those patients with early stage endometrial cancer with both high sensitivity and high specificity (receiver operating characteristics area under the curve = 0.91 for stage 1A and 0.97 for higher than stage 1A). Finally, mice harbouring a Hand2 knock-out specifically in their endometrium were shown to develop precancerous endometrial lesions with increasing age, and these lesions also demonstrated a lack of PTEN expression. Conclusions: HAND2 methylation is a common and crucial molecular alteration in endometrial cancer that could potentially be employed as a biomarker for early detection of endometrial cancer and as a predictor of treatment response. The true clinical utility of HAND2 DNA methylation, however, requires further validation in prospective studies.publishedVersio

A First-Time-in-Human Study of GSK2636771, a Phosphoinositide 3 Kinase Beta-Selective Inhibitor, in Patients with Advanced Solid Tumors.

Background: The PI3K/protein kinase B (AKT) pathway is commonly activated in several tumor types. Selective targeting of p110β could result in successful pathway inhibition while avoiding the on- and off-target effects of pan-PI3K inhibitors. GSK2636771 is a potent, orally bioavailable, adenosine triphosphate-competitive, selective inhibitor of PI3Kβ.Methods: We evaluated the safety, pharmacokinetics, pharmacodynamics and antitumor activity of GSK2636771 to define the recommended phase II dose (RP2D). During the dose-selection and dose-escalation stages (parts 1 and 2), patients with PTEN-deficient advanced solid tumors received escalating doses of GSK2636771 (25-500 mg once daily) using a modified 3+3 design to determine the RP2D; tumor type-specific expansion cohorts (part 3) were implemented to further assess tumor responses at the RP2D.Results: A total of 65 patients were enrolled; dose-limiting toxicities were hypophosphatemia and hypocalcemia. Adverse events included diarrhea (48%), nausea (40%), and vomiting (31%). Single- and repeat-dose exposure increased generally dose proportionally. GSK2636771 400 mg once daily was the RP2D. Phospho/total AKT ratio decreased with GSK2636771 in tumor and surrogate tissue. A castrate-resistant prostate cancer (CRPC) patient harboring PIK3CB amplification had a partial response for over a year; an additional 10 patients derived durable (≥24 weeks) clinical benefit, including two other patients with CRPC with PIK3CB alterations (≥34 weeks). GSK2636771 400 mg once daily orally induced sufficient exposure and target inhibition with a manageable safety profile.Conclusions: Genomic aberrations of PIK3CB may be associated with clinical benefit from GSK2636771. Clin Cancer Res; 23(19); 5981-92. ©2017 AACR

Role of DNA Methylation and Epigenetic Silencing of HAND2 in Endometrial Cancer Development

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