Inhibiting PRMT5 induces DNA damage and increases anti-proliferative activity of Niraparib, a PARP inhibitor, in models of breast and ovarian cancer.

BACKGROUND: Inhibitors of Poly (ADP-Ribose) Polymerases (PARP) provide clinical benefit to patients with breast and ovarian cancers, by compromising the DNA repair activity of cancer cells. Although these agents extend progression-free survival in many patients, responses can be short lived with many patients ultimately progressing. Identification of combination partners that increase dependence of cancer cells to the DNA repair activity of PARPs may represent a strategy to increase the utility of PARP inhibitors. Protein arginine methyltransferase 5 (PRMT5) regulates DNA damage response pathways through splicing and protein modification, and inhibitors of PRMT5 have recently entered clinical trials. METHODS: The effect of PRMT5 inhibition on the levels of DNA damage and repair markers including γH2AX, RAD51, and 53BP1 was determined using high content immunofluorescent imaging. The anti-proliferative activity of the combination of PRMT5 and PARP inhibitors was evaluated using in vitro models of breast and ovarian cancers using both cell lines and ex vivo patient derived xenografts. Finally, the combinations of PRMT5 and PARP inhibitors were evaluated in cell line xenograft models in vivo. RESULTS: Inhibition of PRMT5 by GSK3326595 led to increased levels of markers of DNA damage. The addition of GSK3326595 to the PARP inhibitor, niraparib, resulted in increased growth inhibition of breast and ovarian cancer cell lines and patient derived spheroids. In vivo, the combination improved the partial effects on tumor growth inhibition achieved by either single agent, producing complete tumor stasis and regression. CONCLUSION: These data demonstrate that inhibition of PRMT5 induced signatures of DNA damage in models of breast and ovarian cancer. Furthermore, combination with the PARP inhibitor, Niraparib, resulted in increased anti-tumor activity in vitro and in vivo. Overall, these data suggest inhibition of PRMT5 as a mechanism to broaden and enhance the clinical application of PARP inhibitors.

Therapeutic Targeting of RNA Splicing Catalysis through Inhibition of Protein Arginine Methylation.

Cancer-associated mutations in genes encoding RNA splicing factors (SFs) commonly occur in leukemias, as well as in a variety of solid tumors, and confer dependence on wild-type splicing. These observations have led to clinical efforts to directly inhibit the spliceosome in patients with refractory leukemias. Here, we identify that inhibiting symmetric or asymmetric dimethylation of arginine, mediated by PRMT5 and type I protein arginine methyltransferases (PRMTs), respectively, reduces splicing fidelity and results in preferential killing of SF-mutant leukemias over wild-type counterparts. These data identify genetic subsets of cancer most likely to respond to PRMT inhibition, synergistic effects of combined PRMT5 and type I PRMT inhibition, and a mechanistic basis for the therapeutic efficacy of PRMT inhibition in cancer.

Activation of the p53-MDM4 regulatory axis defines the anti-tumour response to PRMT5 inhibition through its role in regulating cellular splicing.

Evasion of the potent tumour suppressor activity of p53 is one of the hurdles that must be overcome for cancer cells to escape normal regulation of cellular proliferation and survival. In addition to frequent loss of function mutations, p53 wild-type activity can also be suppressed post-translationally through several mechanisms, including the activity of PRMT5. Here we describe broad anti-proliferative activity of potent, selective, reversible inhibitors of protein arginine methyltransferase 5 (PRMT5) including GSK3326595 in human cancer cell lines representing both hematologic and solid malignancies. Interestingly, PRMT5 inhibition activates the p53 pathway via the induction of alternative splicing of MDM4. The MDM4 isoform switch and subsequent p53 activation are critical determinants of the response to PRMT5 inhibition suggesting that the integrity of the p53-MDM4 regulatory axis defines a subset of patients that could benefit from treatment with GSK3326595.

Sun Smart Schools Nevada: Increasing Knowledge Among School Children About Ultraviolet Radiation.

BACKGROUND: Cumulative exposure to ultraviolet radiation (UV) is a risk factor for development of skin cancer. We estimated changes in knowledge, attitudes, and behaviors among Nevada school-age children following implementation of a program to decrease UV exposure. COMMUNITY CONTEXT: The Nevada Cancer Coalition's Sun Smart Schools pilot program was implemented in 7 Nevada schools during the 2015-2016 school year. The target population was students at participating schools. METHODS: Participation in the program was voluntary. Students surveyed spanned grades from fourth to tenth. Pre-intervention surveys were conducted at the start of the school year. Post-intervention surveys were conducted at the end of the school year. Changes in knowledge, attitudes, and behaviors were assessed among study participants by using self-reported survey responses. OUTCOMES: The Sun Smart Schools pilot program was effective in increasing a broad range of knowledge, attitudes, and behaviors about protection from UV among elementary and middle school students. Students in high school reported an increase in the adoption of selected protective behaviors. However, this population also maintained a positive attitude toward the appearance of tanned skin, indicating susceptibility to competing influences. High school students also did not report any evident change in knowledge about sun protection strategies. Parents reported a decrease in knowledge about UV protection but an increase in adoption of certain protective behaviors. INTERPRETATION: Our findings are similar to those of previous studies demonstrating that education about the dangers of UV exposure is most effective in younger age groups. Results were mixed in older age groups.

Validation of the 4 week recall version of the Uterine Fibroid Symptom and Health-related Quality of Life (UFS-QOL) Questionnaire.

OBJECTIVE: To assess the psychometric performance of the 4 week recall version of the Uterine Fibroid Symptom and Health-related Quality of Life Questionnaire (UFS-QoL), a patient measure of the severity of uterine fibroid (UF) symptoms and their impact on health-related quality of life (HRQL). METHODS: This was a retrospective analysis of phase 2a data from pre-menopausal women with heavy menstrual bleeding associated with UF. Participants completed the UFS-QoL at Baseline, Treatment Month 3, and Follow-up Month 3 and a daily diary with a Menstrual Bleeding Scale and the UF Daily Symptom Scale throughout the study duration. Descriptive statistics were performed on patient demographic characteristics; analyses were conducted to assess the internal consistency reliability, validity, and responsiveness of the UFS-QoL 4 week recall version. RESULTS: A total of 271 women were enrolled with a mean age of 41.8 years; 74% were black. The UFS-QoL demonstrated excellent internal consistency reliability, with Cronbach's alpha coefficient values >0.70 for each subscale at each study visit. Results indicated good concurrent validity with the UF Daily Symptom Scale items. The women with amenorrhea at Treatment Month 3 had significantly better scores on all UFS-QoL subscales and HRQL Total than women with menstrual bleeding, indicating acceptable discriminant validity. Mean subscale change scores from Baseline to Treatment Month 3 were 19.2 to 39.8. Effect sizes were moderate to large (0.53 to 1.86), demonstrating responsiveness to change. LIMITATIONS: As this study is a post hoc validation of the 4 week recall UFS-QOL, it is limited to the clinical trial data available and does not include a direct comparison to the 3 month recall version of UFS-QOL. CONCLUSIONS: The 4 week recall version of the UFS-QoL demonstrated good internal consistency reliability, concurrent validity, and responsiveness and is psychometrically comparable to the original 3 month recall UFS-QoL. CLINICAL TRIAL REGISTRATION: Data from a phase 2a, cohort design proof of concept study (trial M12-663); ClinicalTrials.gov identifier NCT01441635. Date of Registration: 6 September 2011.

Responsiveness of the Peyronie's Disease Questionnaire (PDQ).

INTRODUCTION: In order to reliably assess treatment effectiveness, patient-reported outcome instruments must demonstrate adequate psychometric properties. AIM: To assess the responsiveness of the Peyronie's Disease Questionnaire (PDQ) using data from two Phase 3 trials of collagenase clostridium histolyticum for Peyronie's disease (PD). METHODS: Both trials recruited adult males with PD who were in a stable relationship with a female partner for at least 3 months. Patients completed the PDQ, International Index of Erectile Function (IIEF), and a global assessment of PD (GAPD) questionnaire at baseline and Weeks 24 and 52. Anchor- and distribution-based methods were used to evaluate the responsiveness of the PDQ. MAIN OUTCOME MEASURE: Peyronie's Disease Questionnaire. RESULTS: The number of men available with baseline and Week 52 data was 267 for Study 1 and 270 for Study 2. The mean age was 58.0 for Study 1 and 57.4 for Study 2; the majority were white (95.2% and 97.3%, respectively). Mean PDQ subscale change scores from baseline to Week 52 for both studies ranged from -1.5 to -4.6 (P < 0.0001). In Study 1, effect sizes were moderate to large on the Psychological and Physical Symptoms (-0.56) and Symptom Bother subscales (-0.84). For patients with penile pain at baseline, the effect size was large (-1.05) for the Penile Pain subscale. Similar effect sizes were seen in Study 2. The Psychological and Physical Symptoms and Symptom Bother subscales significantly discriminated patient improvement ratings of GAPD and degree of penile curvature at Weeks 24 and 52. CONCLUSIONS: The PDQ is highly responsive to change in men with PD.

The test-retest reliability of the Peyronie's disease questionnaire.

INTRODUCTION: The Peyronie's Disease Questionnaire (PDQ) is a disease-specific, patient-reported outcome instrument designed to measure the psychosexual consequences and treatment outcomes of Peyronie's disease (PD). AIM: The aim of this study was to evaluate the test-retest reliability of the PDQ. METHODS: Adult men with PD were recruited through eight clinical sites across the United States. Participants completed the PDQ during two study visits scheduled 7 (± 3) days apart. At Visit 1, participants completed a sociodemographic questionnaire, the PDQ, and the International Index of Erectile Function (IIEF). At Visit 2, participants repeated the PDQ and completed an Overall Treatment Effect (OTE) scale. Test-retest reliability of the PDQ was assessed in a stable subsample (as determined by responses to the OTE). Intraclass correlation coefficients (ICCs) were calculated to evaluate the degree of association between the three PDQ subscale scores at Visits 1 and 2. Internal consistency of the subscales was also evaluated using Cronbach's alpha. MAIN OUTCOME MEASURE: The main outcome measure was the PDQ. RESULTS: Of the 61 PD patients (mean age 59.3) who took part in the study, the majority were not receiving treatment for their PD (n=35, 57.4%). The sample's mean score on the erectile function domain of IIEF was 19.7 (± 8.2), indicating mild-moderate dysfunction. Nearly two-thirds reported penile pain at baseline (n=37, 63.8%). Of the participants with baseline PDQ data and who had engaged in vaginal intercourse in the past 3 months, 57 completed both study visits. The PDQ demonstrated excellent test-retest reliability in 53 stable patients. The ICC was 0.85 for the Psychological and Physical Symptom subscale, 0.89 for the Peyronie's Symptom Bother subscale, and 0.88 for the Penile Pain subscale. The Cronbach's alpha estimates for all three subscales were acceptable at the >0.70 level. CONCLUSIONS: The PDQ is a highly reproducible measure of PD and can be an effective end point in clinical trials evaluating treatments for PD.

Insights into the binding of PARP inhibitors to the catalytic domain of human tankyrase-2.

The poly(ADP-ribose) polymerase (PARP) family represents a new class of therapeutic targets with diverse potential disease indications. PARP1 and PARP2 inhibitors have been developed for breast and ovarian tumors manifesting double-stranded DNA-repair defects, whereas tankyrase 1 and 2 (TNKS1 and TNKS2, also known as PARP5a and PARP5b, respectively) inhibitors have been developed for tumors with elevated ß-catenin activity. As the clinical relevance of PARP inhibitors continues to be actively explored, there is heightened interest in the design of selective inhibitors based on the detailed structural features of how small-molecule inhibitors bind to each of the PARP family members. Here, the high-resolution crystal structures of the human TNKS2 PARP domain in complex with 16 various PARP inhibitors are reported, including the compounds BSI-201, AZD-2281 and ABT-888, which are currently in Phase 2 or 3 clinical trials. These structures provide insight into the inhibitor-binding modes for the tankyrase PARP domain and valuable information to guide the rational design of future tankyrase-specific inhibitors.

The impact of OAB on physical activity in the United States: results from OAB-POLL.

OBJECTIVE: To provide data on physical activity among those with and without overactive bladder (OAB) in a large, ethnically diverse U.S. sample. METHODS: A cross-sectional survey was conducted via the Internet among 10,000 men and women aged 18-70 (2000 African Americans, 2000 Hispanics, and 6000 whites) using the lower urinary tract symptoms (LUTS) tool and questions from the 2007-2008 National Health and Nutrition Examination Survey (NHANES). OAB cases and those with no/minimal symptoms (NMS) were compared on federal guidelines of indices of physical activity: 2008 guidelines and 2010 Healthy People. Descriptive statistics were used to evaluate differences between OAB and NMS. Logistic regressions examined the impact of OAB on physical activity. RESULTS: Response rate, 57%; 818 men and 1505 women with OAB, and 1857 men and 1615 women with NMS. Respondents with other LUTS were excluded from this analysis (2302 men and 1904 women). Those with OAB were significantly less likely to report moderate and vigorous physical activities in their leisure time and to satisfy recommended physical activity levels compared to those with NMS. Symptoms of OAB (men and women: urgency and urinary frequency; women: urinary urge incontinence) were associated with limitations in physical activity in the logistic regressions. CONCLUSION: This study benchmarks physical activity levels among people with OAB. Men and women with OAB were significantly less likely to achieve recommended physical activity levels than people with NMS. More research is needed to further evaluate how OAB affects physical activity and health status and to determine causal relationships.

Assessment of concomitant testicular dose with radiochromic film.

To assess the suitability of EBT2 and XRQA2 Gafchromic film for measuring low doses in the periphery of treatment fields, and to measure the accumulative concomitant dose to the contralateral testis resulting from CT imaging, pre-treatment imaging (CBCT) and seminoma radiotherapy with and without gonadal shielding. Superficial peripheral dose measurements made using EBT2 Gafchromic film on the surface of water equivalent material were compared to measurements made with an ionisation chamber in a water phantom to evaluate the suitability and accuracy of the film dosimeter for such measurements. Similarly, XRQA2 was used to measure surface doses within a kilovoltage beam and compared with ionisation chamber measurements. Gafchromic film was used to measure CT, CBCT and seminoma treatment related testicular doses on an anthropomorphic phantom. Doses were assessed for two clinical plans, both with and without gonadal shielding. Testicular doses resulting from the treatment of up to 0.83 ± 0.17 Gy were measured per treatment. Additional doses of up to 0.49 ± 0.01 and 2.35 ± 0.05 cGy were measured per CBCT and CT image, respectively. Reductions in the testicular dose in the order of 10, 36 and 78% were observed when gonadal shielding was fitted for treatment, CT and CBCT imaging, respectively. Gafchromic film was found to be suitable for measuring dose in the periphery of treatment fields. The dose to the testis should be limited to minimise the risk of radiation related side effects. This can be achieved by using appropriate gonadal shielding, irrespective of the treatment fields employed.

EZH2 inhibition as a therapeutic strategy for lymphoma with EZH2-activating mutations.

In eukaryotes, post-translational modification of histones is critical for regulation of chromatin structure and gene expression. EZH2 is the catalytic subunit of the polycomb repressive complex 2 (PRC2) and is involved in repressing gene expression through methylation of histone H3 on lysine 27 (H3K27). EZH2 overexpression is implicated in tumorigenesis and correlates with poor prognosis in several tumour types. Additionally, somatic heterozygous mutations of Y641 and A677 residues within the catalytic SET domain of EZH2 occur in diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma. The Y641 residue is the most frequently mutated residue, with up to 22% of germinal centre B-cell DLBCL and follicular lymphoma harbouring mutations at this site. These lymphomas have increased H3K27 tri-methylation (H3K27me3) owing to altered substrate preferences of the mutant enzymes. However, it is unknown whether specific, direct inhibition of EZH2 methyltransferase activity will be effective in treating EZH2 mutant lymphomas. Here we demonstrate that GSK126, a potent, highly selective, S-adenosyl-methionine-competitive, small-molecule inhibitor of EZH2 methyltransferase activity, decreases global H3K27me3 levels and reactivates silenced PRC2 target genes. GSK126 effectively inhibits the proliferation of EZH2 mutant DLBCL cell lines and markedly inhibits the growth of EZH2 mutant DLBCL xenografts in mice. Together, these data demonstrate that pharmacological inhibition of EZH2 activity may provide a promising treatment for EZH2 mutant lymphoma.

Identification of Potent, Selective, Cell-Active Inhibitors of the Histone Lysine Methyltransferase EZH2.

The histone H3-lysine 27 (H3K27) methyltransferase EZH2 plays a critical role in regulating gene expression, and its aberrant activity is linked to the onset and progression of cancer. As part of a drug discovery program targeting EZH2, we have identified highly potent, selective, SAM-competitive, and cell-active EZH2 inhibitors, including GSK926 (3) and GSK343 (6). These compounds are small molecule chemical tools that would be useful to further explore the biology of EZH2.

Sure Start Children's Centres: time to make them better known.

Health services have traditionally been available to local communities through general practices, health visitors and midwives. Since the introduction of the Sure Start Local programme in 1997 and Sure Start Children's Centres in 2004, an increasing number of children's centres are working with their local NHS to provide health services for the under fives, and improve the accessibility of these services to local families. Sure Start Children's Centres were set up in England as part of the Department for Children, Schools and Families' Children's Plan to improve outcomes for children and their families and contribute to the strategic objectives of Every Child Matters. Children's centres offer a range of integrated services from child health and maternity services to parenting and childcare provisions. They also deliver key health promotion activities and programmes that contribute to the public health priorities of Primary Care Trusts, e.g. breast-feeding, smoking in pregnancy, physical activity and healthy eating. There are now 3,500 children's centres offering easily accessible services to more than 2.4 million children and families across England. Health professionals are asked work proactively together to raise awareness of these among local communities so that more families can benefit from the services they offer.

Discovery of GSK1070916, a potent and selective inhibitor of Aurora B/C kinase.

The Aurora kinases play critical roles in the regulation of mitosis and are frequently overexpressed or amplified in human tumors. Selective inhibitors may provide a new therapy for the treatment of tumors with Aurora kinase amplification. Herein we describe our lead optimization efforts within a 7-azaindole-based series culminating in the identification of GSK1070916 (17k). Key to the advancement of the series was the introduction of a 2-aryl group containing a basic amine onto the azaindole leading to significantly improved cellular activity. Compound 17k is a potent and selective ATP-competitive inhibitor of Aurora B and C with K(i)* values of 0.38 +/- 0.29 and 1.5 +/- 0.4 nM, respectively, and is >250-fold selective over Aurora A. Biochemical characterization revealed that compound 17k has an extremely slow dissociation half-life from Aurora B (>480 min), distinguishing it from clinical compounds 1 and 2. In vitro treatment of A549 human lung cancer cells with compound 17k results in a potent antiproliferative effect (EC(50) = 7 nM). Intraperitoneal administration of 17k in mice bearing human tumor xenografts leads to inhibition of histone H3 phosphorylation at serine 10 in human colon cancer (Colo205) and tumor regression in human leukemia (HL-60). Compound 17k is being progressed to human clinical trials.

The responsiveness of the uterine fibroid symptom and health-related quality of life questionnaire (UFS-QOL).

BACKGROUND: A number of noninvasive alternatives to hysterectomy have become available as treatments for uterine fibroids. These alternative therapies, however, may not relieve all symptoms. Consequently, the need for patient-reported outcomes to assess symptom reduction of uterine fibroids has become increasingly important to evaluate the clinical success of patients who choose these alternative therapies. The purpose of the study was to examine the responsiveness of the Uterine Fibroid Symptom and Health-Related Quality of Life Questionnaire (UFS-QOL) with treatment of uterine fibroids. METHODS: The responsiveness of the UFS-QOL was assessed as a post-hoc analysis of patients treated with MRI-guided focused ultrasound thermal ablation (MRgFUS) for uterine fibroids. The UFS-QOL and SF-36 were completed at baseline and months 1, 3, and 6. Patient perceived overall treatment effect (OTE) was assessed at month 3, while satisfaction with treatment was collected at month 6. The responsiveness of the UFS-QOL was examined using effect sizes and change scores by patient-reported overall treatment effect and satisfaction. RESULTS: A total of 102 women with complete UFS-QOL data were included in the analysis; the mean age was 45 years and 79% were Caucasian. From baseline to 6 months, significant improvements were observed in UFS-QOL Symptom Severity and all Health-Related Quality of Life (HRQL) subscale scores (p < 0.0001). When examining change in general health status over the 6-month follow-up period, significant improvements were noted in all 8 SF-36 subscales. The UFS-QOL was highly responsive with subscale effect sizes ranging from 0.74 for Sexual Function to -1.9 for Symptom Severity. Improvements in UFS-QOL subscales were associated with patient perceptions of perceived benefit and treatment satisfaction. CONCLUSION: The UFS-QOL is responsive to treatment for uterine fibroids and is a useful outcome measure for uterine-sparing uterine fibroid treatments.

Validation of the treatment satisfaction questionnaire for Crohn's disease (TSQ-C).

Treatment satisfaction is used to capture the full impact of disease on patients' lives. Currently, no instruments exist to evaluate satisfaction with pharmacologic therapy in patients with Crohn's disease (CD). The purpose of this study was to evaluate the psychometric properties of a treatment satisfaction questionnaire for CD (TSQ-C). The 36-item questionnaire was completed by CD patients who reported taking 5-aminosalicylic acid derivatives to treat their CD. Measures used in the validation study were the Inflammatory Bowel Disease Questionnaire (IBDQ), Crohn's Work Activity Impairment Index (CWAII), and patient reports of clinical indicators (e.g., number of active flares and medications taken). Exploratory factor analysis was used to evaluate the items and subscale structure. Internal consistency reliability and concurrent and discriminant validity were assessed using Cronbach's alpha, Pearson correlation coefficients, and analysis of variance. A total of 813 CD patients participated, with the majority being Caucasian (95.9%), female (67.0%), and >34 years old (86.1%). Patient-rated severity of CD was mild (49.3%), moderate (41.7%), and severe (7.5%). The final TSQ-C consisted of 32 items, with six subscales (Symptoms, Satisfaction, Expectations, Physician Relationships, Bother, and Cost), with each subscale score ranging from 1 to 6. Cronbach's alpha values ranged from 0.63 (Cost) to 0.94 (Symptoms). Strong correlations were observed among the IBDQ, CWAII, and the Satisfaction and Symptoms subscales of the TSQ-C. TSQ-C subscales, particularly Symptoms and Satisfaction, significantly discriminated among levels of number of flares per year, patient-rated disease severity, and number of medication classes. The TSQ-C demonstrated excellent validity and reliability and appears to be a useful tool for evaluating satisfaction with pharmacologic therapy among patients with CD.

Validity and reliability of patient reported outcomes used in psoriasis: results from two randomized clinical trials.

BACKGROUND: Two Phase III randomized controlled clinical trials were conducted to assess the efficacy, safety, and tolerability of weekly subcutaneous administration of efalizumab for the treatment of psoriasis. Patient reported measures of psoriasis-related functionality and health-related quality of life and of psoriasis-related symptom assessments were included as part of the trials. OBJECTIVE: To assess the reliability, validity, and responsiveness of the patient reported outcome measures that were used in the trials - the Dermatology Life Quality Index (DLQI), the Psoriasis Symptom Assessment (PSA) Scale, and two itch measures, a Visual Analog Scale (VAS) and the National Psoriasis Foundation (NPF) itch measure. METHODS: Subjects aged 18 to 70 years with moderate to severe psoriasis for at least 6 months were recruited into the two clinical trials (n = 1095). Internal consistency reliability was evaluated for all patient reported outcomes at baseline and at 12 weeks. Construct validity was evaluated by relations among the different patient reported outcomes and between the patient reported outcomes and the clinical assessments (Psoriasis Area and Severity Index; Overall Lesion Severity Scale; Physician's Global Assessment of Change) assessed at baseline and at 12 weeks, as was the change over the course of the 12 week portion of the trial. RESULTS: Internal consistency reliability ranged from 0.86 to 0.95 for the patient reported outcome measures. The patient reported outcome measures were all shown to have significant construct validity with respect to each other and with respect to the clinical assessments. The four measures also demonstrated significant responsiveness to change in underlying clinical status of the patients over the course of the trial, as measured by the independently assessed clinical outcomes. CONCLUSIONS: The DLQI, the PSA, VAS, and the NPF are considered useful tools for the measurement of dermatology-related limitations of functional ability and the frequency, severity and impact of psoriasis symptoms on patients' lives and psoriasis-related quality of life.