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Differential PARP inhibitor responses in <i>BRCA1</i>-deficient and resistant cells in competitive co-culture

by Shiella A. Soetomo, Michael F. Sharp, Wayne Crismani

Synthetic lethality describes a genetic relationship where the loss of two genes results in cell death, but the loss of one of those genes does not. Drugs used for precision oncology can exploit synthetic lethal relationships; the best described are PARP inhibitors which preferentially kill BRCA1-deficient tumours preferentially over BRCA1-proficient cells. New synthetic lethal targets are often discovered using genetic screens, such as CRISPR knockout screens. Here, we present a competitive co-culture assay that can be used to analyse drugs or gene knockouts with synthetic lethal effects. We generated new BRCA1 isogenic cell line pairs from both a triple-negative breast cancer cell line (SUM149) and adapted pre-existing non-cancerous BRCA1 isogenic pair (RPE). Each cell line of the isogenic pair was transformed with its own fluorescent reporter. The two-coloured cell lines of the isogenic pair were then grown together in the same vessel to create a more competitive environment compared to when grown separately. We used four PARP inhibitors to validate the ability to detect synthetic lethality in BRCA1-deficient cancer cells. The readout of the assay was performed by counting the fluorescently coloured cells after drug treatment using flow cytometry. We observed preferential targeting of BRCA1-deficient cells, by PARPi, at relative concentrations that broadly reflect clinical dosing. Further we reveal subtle differences between PARPi resistant lines compared to BRCA1-proficient cells. Here, we demonstrate the validation and potential use of the competitive assay, which could be extended to validating novel genetic relationships and adapted for live cell imaging.

‘Living Well With a PICC at Home’: Co‐Design and Evaluation of a Peripherally Inserted Central Catheter (PICC) Booklet

ABSTRACT

Aim

To co-design an information booklet to support adults living with a peripherally inserted central catheter (PICC) at home.

Design

A sequential, mixed-method approach using Boyd's co-design framework.

Method

A diverse project team led co-design of a PICC booklet based on preferences and PICC management strategies identified through content analysis of interviews with 15 Australian health consumers (January 2022–March 2023) using a qualitative descriptive approach. A draft booklet was developed, reviewed by the team and prototyped, which was evaluated by consumer participants and an external nurse panel, with readability assessed using Flesch Reading Ease and Flesch–Kincaid scores. The project team reviewed feedback, reached consensus on changes and collaborated with designers to produce the final booklet.

Results

Consumer participant strategies were classified into four categories: enhancing coping, taking responsibility and understanding information, modifying life and accessing supports. Two pre-determined categories were included (participant recommendations for PICC education resource content and top tips for living with a PICC). The prototype was evaluated as highly relevant (3.9/4), comprehensive (3.8/4) with an appropriate writing style and visual appeal. Readability scores were 76.1/100 (Flesch Reading Ease) and 6.9 (Flesch–Kincaid grade level). The final booklet contained 13 sections.

Conclusion

A co-design approach successfully created a novel PICC information booklet with positive evaluations.

Implications

Provides understanding of consumer information preferences to inform PICC education and offers insight into consumer-developed strategies and knowledge for living with a PICC.

Impact

A co-designed PICC information booklet based on consumer knowledge may augment nursing education for people living with a PICC to improve outcomes.

Reporting Method

The qualitative component follows the EQUATOR network COREQ guideline. A co-design research reporting standard is not available.

Patient or Public Contribution

A consumer representative was a member of the project team and contributed to study design, interpretation of findings and development of the booklet and manuscript.

Development of START-EDI guidelines for reporting equality, diversity and inclusion in research: a study protocol

Por: Fadel · M. G. · Kettley-Linsell · H. · Boshier · P. R. · Barnes · R. · Newby · C. · Manyara · A. M. · Buckle · P. · Vyas · D. A. · Hepburn · J. · Edgar-Jones · P. · Rai · T. · Nicholson · B. D. · Cross · A. J. · Sharples · L. D. · Hopewell · S. · Cohen · J. F. · Welch · V. · Bossuyt · P.
Introduction

Acknowledging equality, diversity and inclusion (EDI) in research is not only a moral imperative but also an important step in avoiding bias and ensuring generalisability of results. This protocol describes the development of STAndards for ReporTing EDI (START-EDI) in research, which will provide a set of minimum standards to help researchers improve their consistency, completeness and transparency in EDI reporting. We anticipate that these guidelines will benefit authors, reviewers, editors, funding organisations, healthcare providers, patients and the public.

Methods and analysis

To create START-EDI reporting guidelines, the following five stages are proposed: (i) establish a diverse, multidisciplinary Steering Committee that will lead and coordinate guideline development; (ii) a systematic review to identify the essential principles and methodological approaches for EDI to generate preliminary checklist items; (iii) conduct an international Delphi process to reach a consensus on the checklist items; (iv) finalise the reporting guidelines and create a separate explanation and elaboration document; and (v) broad dissemination and implementation of START-EDI guidelines. We will work with patient and public involvement representatives and under-served groups in research throughout the project stages.

Ethics and dissemination

The study has received ethical approval from the Imperial College London Research Ethics Committee (study ID: 7592283). The reporting guidelines will be published in open access peer-reviewed publications and presented in international conferences, and disseminated through community networks and forums.

Trial registration number

The project is pre-registered within the Open Science Framework (https://osf.io/8udbq/) and the Enhancing the Quality and Transparency of Health Research Network.

Cross-sectional study of the rates of military sexual trauma (MST) and associations with adverse mental health outcomes among UK female ex-service personnel: a study protocol

Por: Obradovic · T. · Murphy · D. · Fear · N. T. · Sharp · M.-L.
Introduction

This study investigates the rates of military sexual trauma (MST) and its associations with adverse mental health among a sample of UK female ex-service personnel who served during the Iraq/Afghanistan eras.

Methods and analysis

Female ex-service personnel, who participated in the fourth phase (Phase 4) of the King’s Centre for Military Health Research (KCMHR) Health and Well-being Cohort Study (2022–2023) and consented to be recontacted for follow-up studies (n=295), are being invited to participate in an online questionnaire between July 2024 and February 2025. The questionnaire contains surveys and questions related to experiences of sexual harassment and sexual assault during and outside of military service, disordered eating and broader female health issues. While the questionnaire relates to several female health topics, this study focuses on the surveys related to experiences of sexual trauma and eating disorders. Sociodemographic variables and some health variables, including post-traumatic stress disorder (PTSD), complex PTSD, common mental disorders, alcohol misuse, physical somatisation and social support, will be extracted from participants’ pre-existing data collected in Phase 4 of the KCMHR Cohort Study. Analyses will assess rates of MST, and hierarchical multiple logistic regressions will investigate associated health impacts. Rates and ORs, employing 95% CIs, will be reported.

Ethics and dissemination

This study has been granted full ethical approval by the King’s College London Research Ethics Committee (Ref: HR/DP-23/24–39040). Participants provide informed consent before participating and have access to a signposting booklet containing contact details for a range of support services. A risk protocol is in place, which outlines the procedure to be undertaken if a participant contacts the research team in distress. Findings will form part of a PhD thesis and will be further disseminated through peer-reviewed publication and dissemination with veteran mental health services and charities, and relevant government departments.

Patient experiences and expectations of faecal immunochemical testing for investigation of colorectal cancer symptoms: a cross-sectional qualitative interview study with patients and practitioners in the UK

Por: Biran · A. · Dobson · C. · Rees · C. J. · Hamilton · W. · Humes · D. · Neilson · L. J. · Turvill · J. · von Wagner · C. · Whelpton · J. · Sharp · L.
Objectives

Faecal immunochemical testing (FIT) is now commonplace in the UK to prioritise symptomatic patients for urgent gastrointestinal investigation. The test requires a stool sample to be collected at home by the patient and returned for analysis. In this qualitative study, we sought to understand the feasibility and acceptability of FIT-based triage for patients.

Design

A cross-sectional, qualitative, experiential interview study.

Setting

Recruitment was through three participating UK NHS sites (Yorkshire, Midlands, North-East). Health professionals were also identified through membership of the BSG/ACPGBI Symptomatic FIT Guideline Development Group and snowball sampling.

Participants

We interviewed 21 patients who had completed FIT and been referred for colonoscopy and 30 primary and secondary care health professionals involved in symptomatic FIT delivery.

Results

Completion of FIT was unproblematic from the perspective of patients who returned the test. However, health professionals expressed concern over non-return. Among patients, understanding of the purpose of FIT and the meaning of results varied. Health professionals acknowledged that ensuring patient understanding of these can be challenging. Patients believed colonoscopy was less likely to miss cancer than FIT. Patients with a family or personal history of cancer were particularly anxious and wanted the reassurance of colonoscopy, even with a negative FIT result.

Conclusions

We found no major barriers to the use of FIT in prioritising symptomatic patients for urgent investigation. Improving communication might increase compliance and, possibly, acceptability of non-referral for colonoscopy in the case of a negative test result.

Influence of cultural beliefs and parental feeding practices on obesity among primary schoolchildren aged 6-12 in Ghana: a qualitative study

Por: Ewusie · T. B. · Coulthard · H. · Sharps · M. · Ochieng · B.

Childhood obesity has become a global public health challenge and as such has attracted worldwide attention due to its negative impact on children’s health. Despite its diverse determinants, there is a paucity of information on cultural beliefs and parental feeding practices related to childhood obesity in Ghana. This study aimed to explore the influence of cultural beliefs and parental feeding practices on obesity among schoolchildren in Ghana.

Background

Childhood obesity is a global public health concern, drawing widespread attention for its negative impact on children’s health. While the determinants are multifaceted, limited information exists on the impact of cultural beliefs and parental feeding practices in the context of childhood obesity in Ghana. The primary objective of this exploratory study was to investigate the influence of cultural beliefs and parental feeding practices on obesity among schoolchildren in Ghana.

Method

Data for the study were collected through an online interview and focus group discussion from a purposively sampled 60 respondents. An audio recording device was used to compile information shared with respondents during the interview and focus group discussion, both held remotely over the internet. Following Braun and Clarke’s procedure for analysing data, audio-recorded information was transcribed verbatim using Microsoft Word. Vital information to address research questions was assigned codes for collation. Similar codes were collated to form subthemes and major themes which aligned with the Attride-Stirling transcription approach of thematic analysis.

Findings

Four themes emerged from data analysis: parental beliefs and perception of weight and feeding practices; evolving dietary practices; the impact of westernisation and socioeconomic status; and lifestyle at home and obesogenic environments. The cultural inclination towards considering obesity as a sign of a ‘well-fed child’ was evident, and traditional feeding practices were found inadequate, necessitating supplementation with modern approaches. Additionally, factors such as digital media, limited playing space and sedentary behaviours facilitated by transportation to school and easy access to electronic devices contributed to obesity among schoolchildren.

Conclusion

While parents actively promoted mixed food diets, this often conflicted with nutritional needs. Parents also inadvertently encouraged sedentary behaviours hindering physical activity and contributing to weight gain among children. The study highlighted the challenges posed by cultural beliefs on body image and modern influences, necessitating a comprehensive understanding to formulate effective interventions to address childhood obesity in the Ghanaian context.

Characterising the volume and variation of multiple urgent suspected cancer referrals in England, April 2013-March 2018: a national cohort study

Por: Roberts · K. · Cooper · N. · Webster · L. · Sharpless · B. · Round · T. · Gildea · C. · Nicholson · B. D.
Objectives

To establish a methodology to categorise urgent suspected cancer (USC) referrals in England and use these categories to understand individual patient referral patterns by demographic characteristics, financial year and referral pathway.

Design

Cross-sectional population-based cohort study.

Setting

From Cancer Waiting Times data, linked to demographic information held by the National Disease Registration Service, referral-level data on all USC referrals in England between 1 April 2013 and 31 March 2018.

Participants

After restricting records to those with an English postcode at referral and with complete demographic information, 9 524 435 referrals were identified for 7 542 592 patients.

Primary and secondary outcome measures

USC referrals were categorised into first and subsequent USC referrals, based primarily on intervals between referral dates. Our primary outcome was to describe the distribution of referral categories by financial year, suspected cancer referral type and four demographic variables. Our secondary aim was to understand which suspected cancer referral types were found in combination within the first 4 months.

Results

During the study period, 7.5 million people had an USC referral, with one in five having more than one referral, with 9.5 million referrals in total. Referrals were categorised as first (91.1%) and subsequent (8.9%) USC referrals. The relative increase in the number of referrals across the study period was largest (78.2%) for subsequent USC referrals.

Subsequent referrals were most common in the gynaecological (10%), lung (10%) and haematological cancer pathways (12%).

Suspected lower gastrointestinal referrals were most frequently included in a pair of USC referrals; it was one of the five most common pairings for 14 out of 16 referral type pathways, contributing to 30% of upper gastrointestinal USC referral pairings.

Conclusion

Multiple USC referrals increased in the study period, particularly within a year of the first referral. Common referral pairings suggest opportunities for pathway reorganisation where common non-specific symptoms result in multiple USC referrals.

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