Search

PDF file, 59K.

PDF file, 3984K, Methylation pattern sequencing analysis of the sFRP1 promoter.

Metastatic solid tumors are aggressive and mostly drug resistant, leading to few treatment options and poor prognosis as seen with clear cell renal cell carcinoma (ccRCC) and triple-negative breast cancer (TNBC). Therefore, the identification of new therapeutic regimes for the treatment of metastatic disease is desirable. ccRCC and TNBC cell lines were treated with the HDAC inhibitor romidepsin and the methyltransferase inhibitor decitabine, two epigenetic modifying drugs approved by the U.S. Food and Drug Administration for the treatment of various hematologic malignancies. Cell proliferation analysis, flow cytometry, quantitative PCR, and immunoblotting techniques were used to evaluate the antitumor synergy of this drug combination and identify the reexpression of epigenetically silenced tumor suppressor genes. Combinatorial treatment of metastatic TNBC and stage IV ccRCC cell lines with romidepsin/decitabine leads to synergistic inhibition of cell growth and induction of apoptosis above levels of individual drug treatments alone. Synergistic reexpression of the tumor suppressor gene secreted frizzled-related protein one (sFRP1) was observed in combinatorial drug-treated groups. Silencing sFRP1 (short hairpin RNA) before combinatorial drug treatment showed that sFRP1 mediates the growth inhibitory and apoptotic activity of combined romidepsin/decitabine. Furthermore, addition of recombinant sFRP1 to ccRCC or TNBC cells inhibits cell growth in a dose-dependent manner through the induction of apoptosis, identifying that epigenetic silencing of sFRP1 contributes to renal and breast cancer cell survival. Combinatorial treatment with romidepsin and decitabine in drug resistant tumors is a promising treatment strategy. Moreover, recombinant sFRP1 may be a novel therapeutic strategy for cancers with suppressed sFRP1 expression. Mol Cancer Ther; 11(10); 2105–15. ©2012 AACR.

PDF file, 130K, sFRP1 re-expression in cancer cells inhibits Wnt signaling.

PDF file, 328K, Authentication of the VHL mutant (Exon 2 c.407T>C) KIJ265T clear cell renal cell carcinoma cell line.

Supplementary Table S1. Cell line authentication by Short Tandem Repeat Profiling (STR) Supplementary Table S2. Full list of genetic variants identified by MSK-IMPACT sequencing Supplementary Table S3. Details for identified high-confidence gene rearrangements resulting in in-frame fusion proteins Supplementary Table S4. Recurrent copy number alterations in thyroid cancer cell lines. Supplementary Table S5. Genes located within the recurrent copy number alteration regions. Supplementary Table S6. Differentially expressed genes, which are located within CNA regions. "Matching_CNA_type" column indicates whether the direction of the expression change corresponds to the CNA type (133 out of 134, overexpression for gene amplification and underexpression for gene deletions). Supplementary Table S7. Thyroid cancer cell lines gene expression data. Normalized gene expression values for all genes in the 44 cell lines assessed by Affymetrix expression microarray in this study ("CU" prefix, University of Colorado). Supplementary Table S8. Genes with differential expression in PTC vs. ATC cell lines. List of differentially expressed genes between PTC-derived vs. ATC-derived cell lines (limma, adjusted p-value

Supplementary Figure S1. Copy number alteration profile for TERT locus in cell lines harboring homozygous TERT promoter mutations. Copy number changes are expressed as shades of red (gain) or blue (loss). Names of cell lines and log-ratio (lr) values are shown. Supplementary Figure S2. Genome-wide copy number alteration profile of PTC-derived cell lines (top) vs. ATC-derived cell lines (bottom). Chromosome numbers are shown on the top panel. Copy number changes are expressed as shades of red (gain) or blue (loss). Supplementary Figure S3. Representation of the chromosomal location for the 16 recurrent copy number alterations identified by GISTIC in 58 thyroid cancer cell lines. Supplementary Figure S4. Hierarchical clustering of gene expression profiles of thyroid cancer cell lines. Spearman correlation distance metric and Ward agglomeration method were used. Colors identify cell lines originating from tumors of different histologic subtypes. Font color indicates the histological type of the primary tumor from which these cell lines were generated, as follows: red= anaplastic; purple= poorly differentiated; green=follicular; blue= papillary thyroid cancer. Colored dots represent driver alteration: blue= BRAFV600E; orange= N-/H-/KRAS; purple= CCDC6-RET; green= NF1; yellow= MKRN1-BRAF; gray= FGFR2-OGDH; black= unknown. Supplementary Figure S5. Expression of individual genes in the thyroid differentiation signature. Supplementary Figure S6. Correlation of BRS calculated using our algorithm vs. the algorithm published by TCGA. The color of the circles indicates tumor genotype: blue- BRAFV600E, red- RAS gene mutation, green - neither BRAFV600E nor RAS mutation, orange - both BRAFV600E and RAS gene mutations present. Tumors with low BRS are BRAFV600E-like; tumors with high BRS are RAS-like. Supplementary Figure S7. Representative pictures of BAM files from MSK-IMPACT sequencing of CUTC5 specimens for three mutations: A. BRAF p.V600E; B. TP53 C135W; and C. ARID1A E1108*. Each panel shows the sequencing reads for the cell line (top) and primary tumor (bottom). The reference allele and protein translation is shown in the bottommost area, whereas the alternative allele is highlighted in the sequencing reads within each panel. Percentages on the left represent the frequency of the alternative allele in each of the samples tested. All three mutations are detected at low frequencies in the original pleural effusion sample, from which CUTC5 cell line was established. Supplementary Figure S8. Evolution of allelic frequencies from thyroid primary tumors to cell lines for A. TERT promoter; and B. TP53. Graphic representation of the alternative allele frequencies ("Alt Allele Freq"). Cell line names are color-coded on the right side of each graph. All TERT promoter mutations are the canonical c.-124C>T or c.-146C>T, whereas TP53 mutations are indicated. Supplementary Figure S9. Genome-wide copy number alteration profile of primary tumor/cell line and/or patient-derived xenograft (PDX) paired samples from 11 patients. Chromosome numbers are shown on the top panel. Copy number changes are expressed as shades of red (gain) or blue (loss). Colored dots indicate specimen types: cell line (green), primary tumor (orange) or PDX (purple). Cell line names are provided. Supplementary Figure S10. Global MSK-IMPACT copy number profile of THJ560 paired samples. Sample-centered representation of the copy number alterations, expressed as log2 ratios, for primary tumor (top panel), cell line (middle panel) and patient-derived xenograft (bottom panel). Table inserts show the top deletions, corresponding to the CDKN2A/CDKN2B locus for the cell line and xenograft, which are absent in the primary tumor.

Supplementary Figure 3:SCD1 inhibition induces ATF6 - PDF file 844K, ATF6 is activated in response to SCD1 inhibition in ccRCC

Supplementary Figure S1 from Cytoplasmic Sequestration of p27 via AKT Phosphorylation in Renal Cell Carcinoma

Supplementary Figure 1: Validation of RWV-366T - PDF file 5154K, Verification of renal origins and confirmation of patient tissue match for RWV-366T ccRCC cell line

Supplementary Methods, Supplementary Figure and Table Legends

Supplementary Figure 5: mono and combo effects of temsirolimus - PDF file 350K, Monotherapeutic Dose out of the mTOR inhibitor temsirolimus in ccRCC cell lines

Supplementary Figure 2: Inhibition of SCD1 induces ER stress - PDF file 1316K, Inhibition of SCD1 activity in ccRCC induces endoplasmic reticulum stress response in ccRCC

Purpose:Thyroid cancer cell lines are valuable models but have been neglected in pancancer genomic studies. Moreover, their misidentification has been a significant problem. We aim to provide a validated dataset for thyroid cancer researchers.Experimental Design:We performed next-generation sequencing (NGS) and analyzed the transcriptome of 60 authenticated thyroid cell lines and compared our findings with the known genomic defects in human thyroid cancers.Results:Unsupervised transcriptomic analysis showed that 94% of thyroid cell lines clustered distinctly from other lineages. Thyroid cancer cell line mutations recapitulate those found in primary tumors (e.g., BRAF, RAS, or gene fusions). Mutations in the TERT promoter (83%) and TP53 (71%) were highly prevalent. There were frequent alterations in PTEN, PIK3CA, and of members of the SWI/SNF chromatin remodeling complex, mismatch repair, cell-cycle checkpoint, and histone methyl- and acetyltransferase functional groups. Copy number alterations (CNA) were more prevalent in cell lines derived from advanced versus differentiated cancers, as reported in primary tumors, although the precise CNAs were only partially recapitulated. Transcriptomic analysis showed that all cell lines were profoundly dedifferentiated, regardless of their derivation, making them good models for advanced disease. However, they maintained the BRAFV600E versus RAS-dependent consequences on MAPK transcriptional output, which correlated with differential sensitivity to MEK inhibitors. Paired primary tumor-cell line samples showed high concordance of mutations. Complete loss of p53 function in TP53 heterozygous tumors was the most prominent event selected during in vitro immortalization.Conclusions:This cell line resource will help inform future preclinical studies exploring tumor-specific dependencies.

Supplementary Figure Legend - PDF file 92K, Figure legends for Supplementary Figures

Purpose: We set out to identify Stearoyl-CoA desaturase 1 (SCD1) as a novel molecular target in clear cell renal cell carcinoma (ccRCC) and examine its role in tumor cell growth and viability in vitro and in vivo independently as well as in combination with current U.S. Food and Drug Administration (FDA)-approved regimens.Experimental Design: Patient normal and ccRCC tissue samples and cell lines were examined for SCD1 expression. Genetic knockdown models and targeted inhibition of SCD1 through use of a small molecule inhibitor, A939572, were analyzed for growth, apoptosis, and alterations in gene expression using gene array analysis. Therapeutic models of synergy were evaluated utilizing pharmacologic inhibition of SCD1 with the tyrosine kinase inhibitors (TKI) sunitinib and pazopanib, and the mTOR inhibitor temsirolimus.Results: Our studies identify increased SCD1 expression in all stages of ccRCC. Both genetic knockdown and pharmacologic inhibition of SCD1 decreased tumor cell proliferation and induced apoptosis in vitro and in vivo. Upon gene array, quantitative real-time PCR, and protein analysis of A939572-treated or SCD1 lentiviral knockdown samples, induction of endoplasmic reticulum stress response signaling was observed, providing mechanistic insight for SCD1 activity in ccRCC. Furthermore, combinatorial application of A939572 with temsirolimus synergistically inhibited tumor growth in vitro and in vivo.Conclusions: Increased SCD1 expression supports ccRCC viability and therefore we propose it as a novel molecular target for therapy either independently or in combination with an mTOR inhibitor for patients whose disease cannot be remedied with surgical intervention, such as in cases of advanced or metastatic disease. Clin Cancer Res; 19(9); 2368–80. ©2013 AACR.

Supplementary Data from Stearoyl-CoA Desaturase 1 Is a Novel Molecular Therapeutic Target for Clear Cell Renal Cell Carcinoma

Supplementary Figure 4: mono and combo effects of pazopanic and sunitinib - PDF file 1678K, Monotherapeutic dose out of pazopanib and sunitinib, and combinatorial effects with SCD1 inhibitor in ccRCC cells

Purpose: p27 localization and expression has prognostic and predictive value in cancer. Little is known regarding expression patterns of p27 in renal cell carcinoma (RCC) or how p27 participates in disease progression or response to therapy.Experimental Design: RCC-derived cell lines, primary tumors, and normal renal epithelial cells were analyzed for p27 expression, phosphorylation (T157 of the NLS), and subcellular localization. RCC-derived cell lines were treated with phosphatidylinositol 3-kinase (PI3K) and mammalian target of rapamycin (mTOR) inhibitors and effects on p27 localization were assessed. The potential contribution of cytoplasmic p27 to resistance to apoptosis was also evaluated.Results: p27 was elevated in tumors compared with matched controls, and cytoplasmic mislocalization of p27 was associated with increasing tumor grade. Cytoplasmic localization of p27 correlated with phosphorylation at T157, an AKT phosphorylation site in the p27 NLS. In RCC cell lines, activated PI3K/AKT signaling was accompanied by mislocalization of p27. AKT activation and phosphorylation of p27 was associated with resistance to apoptosis, and small interfering RNA knockdown of p27 or relocalization to the nucleus increased apoptosis in RCC cells. Treatment with the PI3K inhibitors LY294002 or wortmannin resulted in nuclear relocalization of p27, whereas mTOR inhibition by rapamycin did not.Conclusions: In RCC, p27 is phosphorylated at T157 of the NLS, with increasing tumor grade associated with cytoplasmic p27. PI3K inhibition (which reduces AKT activity) reduces T157 phosphorylation and induces nuclear relocalization of p27, whereas mTOR inhibition does not. Clinical testing of these findings may provide a rational approach for use of mTOR and PI3K/AKT pathway inhibitors in patients with RCC.

Supplementary Figure 4. Measurement of intracellular calcium influx into (A) empty vector or (B) GluR4 transfected Caki1 cells treated with 1ng/µL R-NPTX2 and DMSO or CFM-2 (10µM) over time using the cell permeable fluorescently labeled calcium indicator Calcium GreenTM-1, AM. Results are calculated as change in fluorescence over time (min). (*) Denotes significant change in fluorescence as compared to DMSO control per time point, and (**) denotes a significant change between R-NPTX2 +/- CFM-2 treated cells. (C) Intracellular Ca2+ levels of NPTX2 and NPTX2-GluR4 transfected Caki1 cells pretreated for 1 hour with DMSO or specified doses of CFM-2. (*) Denotes a significant change in fluorescence of NPTX2 and NPTX2-GluR4 as compared to EV control, and (**) indicates significant changes in fluorescence between DMSO and CFM-2 treated groups

Anaplastic thyroid carcinoma (ATC) is a highly aggressive form of the disease for which new therapeutic options are desperately needed. Previously, we showed that the high-affinity peroxisome proliferator–activated receptor γ (PPARγ) agonist, RS5444, inhibits cell proliferation of ATC cells via induction of the cyclin-dependent kinase inhibitor p21WAF1/CIP1 (p21). We show here that up-regulation of RhoB is a critical step in PPARγ-mediated activation of p21-induced cell stasis. Using multiple independently derived ATC cell lines, we found that treatment with RS5444 leads to the up-regulation of RhoB and subsequent activation of p21, and that silencing of RhoB by RNAi blocks the ability of RS5444 to induce p21 and to inhibit cell proliferation. Our results show that transcriptional regulation of RhoB by the nuclear transcription factor PPARγ is responsible for the induction of p21 mRNA and protein. We further implicate RhoB as a key signaling effector for the growth inhibition of ATC, as treatment with a histone deacetylase inhibitor shown to increase RhoB expression in lung cancer cells caused the up-regulation of RhoB in ATC cells accompanied by increased expression of p21 and inhibition of cell proliferation; this effect occurred even in ATC cells that were unresponsive to RS5444 due to a lack of expression of PPARγ. Our results implicate RhoB as a novel intermediate in critical signaling pathways and as an additional target for therapeutic intervention in ATC. [Cancer Res 2009;69(4):1536–44]

Supplementary Figure 3 from Reactivation of Suppressed RhoB is a Critical Step for the Inhibition of Anaplastic Thyroid Cancer Growth

Supplementary Figure Legends 1-3 from Reactivation of Suppressed RhoB is a Critical Step for the Inhibition of Anaplastic Thyroid Cancer Growth

Supplementary Figure 1 from Reactivation of Suppressed RhoB is a Critical Step for the Inhibition of Anaplastic Thyroid Cancer Growth

Supplementary Figure 2 from Reactivation of Suppressed RhoB is a Critical Step for the Inhibition of Anaplastic Thyroid Cancer Growth

Objectives This study assessed preferences for human papillomavirus (HPV) self-sampling if offered as an alternative to clinician-based screening at the point of invitation for cervical screening. Setting and Methods An online questionnaire was completed by screening-eligible women living in England (n = 3672). Logistic regressions explored associations between demographic characteristics and screening preferences, stratified by previous screening attendance. Reasons for preferences were also assessed. Results Half of participants (51.4%) intended to choose self-sampling, 36.5% preferred clinician screening, 10.5% were unsure, and Conclusions If offered a choice of screening, around half of women in England may choose self-sampling, but a substantial proportion would still opt for clinician screening. Screening providers will need to manage a high take-up of self-sampling if many regular attenders switch to self-sampling.

BackgroundCervical cancer disproportionately affects women ≥65 years, especially those not screened regularly. Speculum use is a key barrier.AimTo assess if offering non-speculum clinician-taken sampling and self-sampling increases uptake for lapsed attenders aged 50–64 years.Design and settingPragmatic randomised control trial conducted at 10 general practices in East London, UK.MethodParticipants were 784 women aged 50–64 years, last screened 6–15 years before randomisation. Intervention participants received a letter offering the choice of non-speculum clinician- or self-sampling. Control participants received usual care. The main outcome measure was uptake within 4 months.ResultsScreening uptake 4 months after randomisation was significantly higher in the intervention arm: 20.4% (n = 80/393) versus 4.9% in the control arm (n = 19/391, absolute difference 15.5%, 95% confidence interval [CI] = 11.0% to 20.0%, Pn = 120/393) versus control 13.6% (n = 53/391) (absolute difference 17.0%, 95% CI = 11.3% to 22.7%, Pn = 50/393) and control 13.6% (n = 53/391) arms. Ethnic differences were seen in screening modality preference. More White women opted for self-sampling (50.7%, n = 38/75), whereas most Asian and Black women and those from other ethnic backgrounds opted for conventional screening.ConclusionOffering non-speculum clinician-taken sampling and self-sampling substantially increases uptake in older lapsed attendee women. Non-speculum clinician sampling appeals to women who dislike the speculum but still prefer a clinician to take their sample. Providing a choice of screening modality may be important for optimising cervical screening uptake.

Objectives HPV vaccination is highly effective in preventing HPV-associated disease, including cervical cancer, which disproportionately affects women from disadvantaged and minority ethnic backgrounds. We examined inequalities in initiation of the HPV vaccination schedule among young women in the UK and reasons given by their parents for non-initiation. Design Cross sectional analyses of a prospective nationally representative cohort study. Setting Four UK countries. Participants 5,695 young women (39.9% from households in lowest income quintiles, 5.1% ever excluded from school, 0.5% not attending school) whose parents (14.3% from minority ethnic backgrounds; 54.1% with no stated religious faith) took part in interviews conducted when their daughters were 14 years old. Main outcome measures Parent-reported initiation of HPV vaccination and reasons for non-initiation. The adjusted odds (aORs) and 95% Confidence Intervals (CI) of initiating HPV vaccination were estimated using logistic regression after mutual adjustment for household income, school exclusion, school attendance and parental ethnic background and religious faith. Results 92.3% (5265) had initiated HPV vaccination at time of interview. Initiation was less likely among those living in the poorest households (aOR; 95% CI: 0.44; 0.30 to 0.64 for those in lowest household income quintile), who did not attend school (0.11; 0.04 to 0.33), had ever been excluded from school (0.47; 0.29 to 0.76), or whose parents were from Black African (0.49; 0.26 to 0.95) or Any Other (0.34; 0.17 to 0.66) ethnic backgrounds. A reason consistent with a conscious or practical decision was reported by 53.3% (219) and 24.1% (90) parents respectively. Conclusions Although most young women are immunised, marked social inequalities in access to HPV vaccination initiation remain. Practical steps to address this are possible and should be implemented to reduce inequalities in primary prevention of cancers and to ensure equitable access to this important public health intervention.

Over 20% of women aged 50-64 in Britain have not attended cervical screening within the recommended 5-year interval. The aim of the present study was to investigate the impact of five messages, informed using strategies from the Behaviour Change Wheel, on strength of intention to attend cervical screening in women aged 50-64 with weak positive intentions to be screened when next invited. Women were randomised (2:2:1), into one of two intervention groups or a control group. The control group saw basic information about cervical screening. Intervention group 1 saw a social norms message and an outcome expectancy message. Intervention group 2 saw a risk reduction message and a response efficacy message. There was further randomisation within the two intervention groups (1:1) to test the effectiveness of message framing and age-targeted information. Lastly, both intervention groups were randomised (1:1) to see a message acknowledging the possible discomfort associated with screening and offering support, or the support message only. Data were included from 475 women, collected using an online survey in March 2022. Adjusting for baseline intention, social norms (p = .84), outcome expectancy (p = .51), risk reduction (p = .19), response efficacy (p = .23) and discomfort acknowledgement messages (p = .71) had no effect on intention strength. However, there was a significant increase in intention after reading multiple messages. These results suggest that although no single message has a significant impact on intentions, when combined, they may act together to increase intention strength. Further research will understand the impact of these messages when combined in information materials.

OBJECTIVES Low uptake of cervical screening in women in their 50s and 60s leaves them at elevated risk of cancer in older age. An age-targeted intervention could be an effective way to motivate older women to attend cervical screening. Our primary objective was to test the impact of different candidate messages on cervical screening intention strength. DESIGN A cross-sectional online survey with randomized exposure to different candidate messages. METHODS Women aged 50-64 years who were not intending to be screened when next invited were recruited through an online panel. Those meeting the inclusion criteria (n = 825) were randomized to one of three groups: (1) control group, (2) intervention group 1, (3) intervention group 2. Each intervention group saw three candidate messages. These included a descriptive social norms message, a diagram illustrating the likelihood of each possible screening outcome, a response efficacy message, a risk reduction message and an acknowledgement of the potential for screening discomfort. We tested age-targeted versions (vs. generic) of some messages. The primary outcome was screening intention strength. RESULTS After adjusting for baseline intention, social norms (p = .425), outcome expectancy (p = .367), risk reduction (p = .090), response efficacy (p = .136) and discomfort acknowledgement messages (p = .181) had no effect on intention strength. Age-targeted messages did not result in greater intention than generic ones. CONCLUSIONS There was no evidence that a single message used to convey social norms, outcome expectancy, risk reduction or response efficacy had an impact on intention strength for older women who did not plan to be screened in future.

OBJECTIVES: To examine the psychosexual impact and disclosure experiences of women testing HPV-positive following cervical screening.DESIGN: In-depth semi-structured interviews.METHODS: Interviews were conducted with 21 women of screening age (i.e. those aged 24-65 years) in England who self-reported testing HPV-positive in the context of cervical screening in the last 12 months. Data were analysed using Framework Analysis.RESULTS: The sexually transmitted nature of HPV, and aspects relating to the transmission of HPV and where their HPV infection had come from, had an impact on women's current, past and future interpersonal and sexual relationships. Most women had disclosed their HPV infection to others, however the factors influencing their decision, and others' reactions to disclosure differed. The magnitude and extent of psychosexual impact was influenced by how women conceptualized HPV, their understanding of key aspects of the virus, concerns about transmitting HPV and having a persistent HPV infection.CONCLUSIONS: Increasing knowledge of key aspects of HPV, such as its high prevalence and spontaneous clearance, and the differences between HPV and other STIs, may increase women's understanding of their screening result and reduce any negative psychosexual consequences of testing HPV-positive. Referring to HPV as an infection that is passed on by skin-to-skin contact during sexual activity, rather than an STI, may help to lessen any psychosexual impact triggered by the STI label.

Objective To explore reasons for variations in anxiety in women testing positive for human papillomavirus (HPV) with normal cytology at routine HPV primary cervical cancer screening. Methods In‐depth interviews were conducted with 30 women who had tested HPV‐positive with normal cytology, including 15 with low‐to‐normal anxiety and 15 with high anxiety. Data were analysed using Framework Analysis to compare themes between low and high anxiety groups. Results Several HPV‐related themes were shared across anxiety groups, but only highly anxious women expressed fear and worry, fatalistic cognitions about cancer, fertility‐related cognitions, adverse physiological responses and changes in health behaviour(s). In comparison to those with low anxiety, women with high anxiety more strongly voiced cognitions about the 12‐month wait for follow‐up screening, relationship infidelity, a lower internal locus of control and HPV‐related symptom attributions. Conclusions Receiving an HPV‐positive with normal cytology result related to various emotional, cognitive, behavioural and physiological responses; some of which were specific to, or more pronounced in, women with high anxiety. If our observations are confirmed in hypothesis‐driven quantitative studies, the identification of distinct themes relevant to women experiencing high anxiety can inform targeted patient communications and HPV primary screening implementation policy., Highlights To our knowledge, this is the first qualitative study to explore anxiety in women testing human papillomavirus (HPV)‐positive with normal cytology at routine HPV primary screening.Our comparative qualitative design allowed thematic nuances to emerge between women who had experienced low versus high anxiety following their result.Only highly anxious women expressed fear and worry, fatalistic cognitions about cancer, fertility‐related cognitions, adverse physiological responses and changes in health behaviour(s).Highly anxious women also more strongly voiced cognitions about the 12‐month wait for follow‐up screen, reltionship infidelity, a lower internal locus of control and HPV‐related symptom attributions.These distinct themes can be used to minimise unnecessary anxiety through the development of evidence‐based patient communications at HPV primary screening.

BackgroundHuman papillomavirus (HPV)-based cervical screening is now replacing cytology-based screening in several countries and many women in screening programmes will consequently receive HPV-positive results. Because of the sexually transmitted nature of HPV, receiving an HPV-positive result may raise questions about disclosing the infection to a sexual partner.ObjectiveTo review the quantitative and qualitative literature exploring women’s concerns about disclosing a high-risk cervical HPV infection to a sexual partner.MethodsWe searched MEDLINE, PsycINFO, CINAHL Plus, Web of Science and EMBASE for studies reporting at least one disclosure-related outcome among women with high-risk HPV. We also searched the grey literature and carried out forward/backward citation searches. A narrative synthesis for quantitative studies and a thematic synthesis for qualitative studies were conducted.ResultsThirteen articles met the inclusion criteria (12 qualitative, 1 quantitative). In the quantitative study, 60% of HPV-positive women felt disclosing an HPV result was ‘risky’. Concerns about disclosing HPV to a sexual partner were influenced by the stigma that is associated with having an STI and uncertainty about how their partner would respond. Women questioned how, when and to whom they should disclose their HPV-positive status.ConclusionsThe studies included in this review provide rich information about the range of concerns women have, the reasons for these concerns, and the questions women have about disclosing HPV to sexual partners. As studies were predominantly qualitative, the prevalence of concerns is unclear.

Highlights • Awareness of HPV is only just over 50% in parents whose children will soon be eligible for vaccination. • More parents of girls than boys said they would vaccinate their child against HPV. • Fathers, and parents of boys were more likely to be undecided about HPV vaccination. • Potentially modifiable HPV vaccine attitudes were strong predictors of intention to vaccinate., Background School-based HPV vaccination in the UK will soon be extended to boys. Based on other countries’ experience, uptake may initially be lower in boys than girls. We assessed HPV vaccine attitudes and decision-making in parents of boys and girls, to explore sex differences and inform public health messages. Methods We carried out a cross-sectional population-based survey using home-based interviews in spring 2019. Participants were adults in England and Wales, with a child in school years 5–7 (aged 9–12 and eligible for HPV vaccination within 3 years). Measures included awareness of HPV and the vaccine, demographic factors, previous vaccine refusal and (after exposure to brief information) whether participants would allow their child to have the HPV vaccine (decided to vaccinate; decided not to vaccinate; undecided). We also assessed vaccine attitudes. Data were weighted to adjust for non-response. Multinomial logistic regression was used to explore predictors of deciding to (or not to) vaccinate compared with being undecided. Results Among 1049 parents (weighted n = 1156), 55% were aware of HPV and the girls’ vaccination programme, but only 23% had heard of plans to vaccinate boys. After information exposure, 62% said they would vaccinate their child, 10% would not, and 28% were undecided. Parents of girls were more willing to vaccinate than parents of boys (adjusted odds ratio: 1.80 (1.32–2.45)). Positive attitudes and HPV/vaccine awareness were significantly independently associated with deciding to vaccinate. Previous vaccine refusal for a child was the strongest predictor of not wanting the HPV vaccine. Conclusions Our findings suggest a need for public health campaigns to raise awareness of plans to extend HPV vaccination to boys. Reassuringly only 10% of all parents were unwilling to vaccinate and our data suggest further information, including about safety and efficacy, may be important in supporting undecided parents to make the decision to vaccinate.

ObjectivesThe introduction of primary Human Papillomavirus (HPV) testing in the National Health Service (NHS) Cervical Screening Programme in England means the screening interval for 25–49 years can be extended from 3 to 5 years. We explored women’s responses to the proposed interval extension.MethodsWe conducted semi-structured phone/video interviews with 22 women aged 25–49 years. Participants were selected to vary in age, socioeconomics and screening history. We explored attitudes to the current 3-year interval, then acceptability of a 5-year interval. Interviews were transcribed verbatim and analysed using framework analysis.ResultsAttitudes to the current 3-year interval varied; some wanted more frequent screening, believing cancer develops quickly. Some participants worried about the proposed change; others trusted it was evidence based. Frequent questions concerned the rationale and safety of longer intervals, speed of cancer development, the possibility of HPV being missed or cell changes occurring between screens. Many participants felt reassured when the interval change was explained alongside the move to HPV primary screening, of which most had previously been unaware.ConclusionsCommunication of the interval change should be done in the context of broader information about HPV primary screening, emphasising that people who test negative for HPV are at lower risk of cell changes so can safely be screened every 5 years. The long time needed for HPV to develop into cervical cancer provides reassurance about safety, but it is important to be transparent that no screening test is perfect.

Background Systematic reviews have identified effective strategies for increasing postal response rates to questionnaires; however, most studies have isolated single techniques, testing the effect of each one individually. Despite providing insight into explanatory mechanisms, this approach lacks ecological validity, given that multiple techniques are often combined in routine practice. Methods We used a two-armed parallel randomised controlled trial (n = 2702), nested within a cross-sectional health survey study, to evaluate whether using a pragmatic combination of behavioural science and evidenced-based techniques (e.g., personalisation, social norms messaging) in a study invitation letter increased response to the survey, when compared with a standard invitation letter. Participants and outcome assessors were blinded to group assignment. We tested this in a sample of women testing positive for human papillomavirus (HPV) at cervical cancer screening in England. Results Overall, 646 participants responded to the survey (response rate [RR] = 23.9%). Logistic regression revealed higher odds of response in the intervention arm (n = 357/1353, RR = 26.4%) compared with the control arm (n = 289/1349, RR = 21.4%), while adjusting for age, deprivation, clinical site, and clinical test result (aOR = 1.30, 95% CI: 1.09–1.55). Conclusion Applying easy-to-implement behavioural science and evidence-based methods to routine invitation letters improved postal response to a health-related survey, whilst adjusting for demographic characteristics. Our findings provide support for the pragmatic adoption of combined techniques in routine research to increase response to postal surveys. Trial registration ISRCTN, ISRCTN15113095. Registered 7 May 2019 – retrospectively registered.

Western blot processing is a well-established procedure that includes protein extraction from tissues and cells, gel electrophoresis separation, transfer to a membrane, and immunodetection with specific antibodies. Here, we show that optimization of washing helps to maximize the specific interactions of antigens and antibodies. Performing all washing steps at 4 °C ensures a maximal signal to noise ratio and reduces nonspecific signals.

Western blot processing is a well-established procedure that includes protein extraction from tissues and cells, gel electrophoresis separation, transfer to a membrane, and immunodetection with specific antibodies. Here, we show that optimization of washing helps to maximize the specific interactions of antigens and antibodies. Performing all washing steps at 4 °C ensures a maximal signal to noise ratio and reduces nonspecific signals.

This study explored knowledge of cervical cancer risk factors among cervical screening non-participants in Great Britain. The aim was to identify knowledge gaps that could be targeted in screening information materials or public education campaigns. We used a cross-sectional design to survey women aged 25 to 64 years living in Great Britain, identified as cervical screening non-participants through self-report questions. Data were collected via a household survey. Survey questions measured awareness of risk factors for cervical cancer and socio-demographic factors. Screening non-participants were included in the study (n = 793) and classified into non-participant groups based on the Precaution Adoption Process Model. Across the sample, 57% of participants identified ‘not going for regular smear tests’ as a risk factor for cervical cancer. Women who intended to be screened were more likely to identify this risk factor than other non-participant groups (OR = 2.13, 95% CI: 1.51–2.99). Women age 55–64 years (OR = 0.60, 95% CI: 0.39–0.93) and women from non-white ethnic backgrounds (OR = 0.70, 95% CI: 0.52–0.94) were less likely to recognise this risk factor. Recognition was lower for ‘infection with human papillomavirus’ (41%). Just over half the sample were aware that screening non-attendance is associated with increased cervical cancer risk, suggesting that non–attendance at screening is not always based on an accurate understanding of the offer. Overall, non-participants are poorly informed about cervical cancer risk factors and further work is needed to ensure that women are making informed choices about (non-) participation., Highlights • Awareness of cervical cancer risk factors is low among screening non-participants. • 41% of cervical screening non-participants knew that HPV is a risk factor. • Older women are less likely to recognise screening non-attendance as a risk factor.

ObjectivesUptake of cervical screening among women aged 50–64 years is declining. Not feeling at risk because of current sexual behaviour is a reason some older women give for not being screened. We hypothesised that explaining the long interval between acquiring human papillomavirus (HPV) and developing cervical cancer would increase the relevance of screening in older women.MethodsWomen aged 50–64 years (n=597) who did not intend to go for screening were recruited through an online panel and randomised to one of three information conditions: cause only (basic information about HPV and cervical cancer), cause with basic timeline (also read a sentence describing the long interval between acquiring HPV and developing cervical cancer) and cause with explicit timeline (read the same as the timeline group alongside an explanation of what this means for older women). Perceived risk of cervical cancer, screening intention strength and understanding of HPV were assessed preinformation and postinformation exposure.ResultsInformation condition was significantly associated with risk perceptions and intention strength postintervention (F(2,593)=6.26, p=0.002 and F(2,593)=4.98, p=0.007 respectively). Women in the cause with explicit timeline condition were more likely to increase their risk perceptions and intention strength compared with cause only (24% vs 9% and 25% vs 13% for risk perceptions and intention, respectively). In the cause with explicit timeline group, women with 4–10 lifetime partners had higher odds of increasing their perceived risk and intention strength postintervention compared with those with 0–1 partners (OR=2.27, 95% CI 1.01 to 5.12 and OR=3.20, 95% CI 1.34 to 7.67, respectively).ConclusionsProviding a clear explanation that decouples women’s perceived cervical cancer risk from their current sexual behaviour has the potential to increase perceived risk of cervical cancer and intentions to be screened among older women. Providing women with a clear cognitive representation of the aetiology of cervical cancer may be one approach to increasing screening uptake.

ObjectivesMany countries are implementing human papillomavirus (HPV)‐based cervical screening due to the higher sensitivity of the test compared with cytology. As HPV is sexually transmitted, there may be psychosexual consequences of testing positive for the virus. We aimed to review the literature exploring the psychosexual impact of testing positive for high‐risk cervical HPV.MethodsMEDLINE, PsycINFO, CINAHL Plus, Web of Science, and EMBASE were searched with no date limits. We also searched the grey literature, reference lists of included articles and carried out forward citation searching. Eligible studies reported at least one psychosexual outcome among HPV‐positive women. Qualitative and quantitative papers were included. We extracted data using a standardised form and carried out a quality assessment for each article. We conducted a narrative synthesis for quantitative studies and a thematic synthesis for qualitative studies.ResultsTwenty‐five articles were included. Quantitative study designs were diverse making it difficult to determine the impact that an HPV positive result would have in the context of routine screening. The qualitative literature suggested that psychosexual concerns cover a broad range of aspects relating to women's current and past relationships, both interpersonal and sexual.ConclusionsThe psychosexual impact of testing positive for high‐risk cervical HPV is unclear. This review highlights the need for further research in the context of HPV‐based cervical screening. As primary HPV testing is introduced more widely, it is important to understand women's responses to testing HPV positive in the cancer screening context to minimise any adverse psychosexual impact.

Purpose: Thyroid cancer cell lines are valuable models but have been neglected in pancancer genomic studies. Moreover, their misidentification has been a significant problem. We aim to provide a validated dataset for thyroid cancer researchers. Experimental Design: We performed next-generation sequencing (NGS) and analyzed the transcriptome of 60 authenticated thyroid cell lines and compared our findings with the known genomic defects in human thyroid cancers. Results: Unsupervised transcriptomic analysis showed that 94% of thyroid cell lines clustered distinctly from other lineages. Thyroid cancer cell line mutations recapitulate those found in primary tumors (e.g., BRAF, RAS, or gene fusions). Mutations in the TERT promoter (83%) and TP53 (71%) were highly prevalent. There were frequent alterations in PTEN, PIK3CA, and of members of the SWI/SNF chromatin remodeling complex, mismatch repair, cell-cycle checkpoint, and histone methyl- and acetyltransferase functional groups. Copy number alterations (CNA) were more prevalent in cell lines derived from advanced versus differentiated cancers, as reported in primary tumors, although the precise CNAs were only partially recapitulated. Transcriptomic analysis showed that all cell lines were profoundly dedifferentiated, regardless of their derivation, making them good models for advanced disease. However, they maintained the BRAFV600E versus RAS-dependent consequences on MAPK transcriptional output, which correlated with differential sensitivity to MEK inhibitors. Paired primary tumor-cell line samples showed high concordance of mutations. Complete loss of p53 function in TP53 heterozygous tumors was the most prominent event selected during in vitro immortalization. Conclusions: This cell line resource will help inform future preclinical studies exploring tumor-specific dependencies.