Investigation of CFTR Function in Human Nasal Epithelial Cells Informs Personalized Medicine.

We broaden the clinical versatility of human nasal epithelial (HNE) cells. HNEs were isolated from 10 participants harboring CFTR variants: nine with rare variants (Q359R [n=2], G480S, R334W [n=5], and R560T) and one person harboring R117H;7T;TG10/5T;TG12. Cultures were differentiated at air-liquid interface. CFTR function was measured in Ussing chambers at three conditions - baseline, ivacaftor, and elexacaftor+tezacaftor+ivacaftor (ETI). Four participants initiated modulators. Q359R HNEs had 5.4% (%WT) baseline CFTR function and 25.5% with ivacaftor. With therapy, sweat [Cl-] decreased and symptoms resolved. G480S HNEs had 4.1% baseline and 32.1% CFTR function with ETI. Clinically, FEV1 increased and sweat [Cl-] decreased (119 to 46mmol/L) with ETI. In vitro cultures derived from five individuals harboring R334W showed a moderate increase in CFTR function with exposure to modulators. For one of these participants, ETI was begun in vivo; symptoms and FEV1 improved. c.1679G>C (R560T) HNEs had <4% baseline CFTR function and no modulator response. RNA analysis confirmed that c.1679G>C completely mis-splices. A symptomatic patient harboring R117H;7T;TG10/5T;TG12 exhibited reduced CFTR function (17.5%) in HNEs, facilitating mild CF diagnosis. HNEs responded to modulators (ivacaftor: 32.8%, ETI: 55.5%) and, since beginning therapy, lung function improved. While reaffirming HNE use for guiding therapeutic approaches, we inform predictions on modulator response (e.g. R334W) and closely assess variants affecting splicing (e.g. c.1679G>C). Notably, functional studies in HNEs harboring R117H;7T;TG10/5T;TG12 facilitated mild CF diagnosis, suggesting use for HNE functional studies as a clinical diagnostic test.

Protospacer modification improves base editing of a canonical splice site variant and recovery of CFTR function in human airway epithelial cells.

Canonical splice site variants affecting the 5' GT and 3' AG nucleotides of introns result in severe missplicing and account for about 10% of disease-causing genomic alterations. Treatment of such variants has proven challenging due to the unstable mRNA or protein isoforms that typically result from disruption of these sites. Here, we investigate CRISPR-Cas9-mediated adenine base editing for such variants in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. We validate a CFTR expression minigene (EMG) system for testing base editing designs for two different targets. We then use the EMG system to test non-standard single-guide RNAs with either shortened or lengthened protospacers to correct the most common cystic fibrosis-causing variant in individuals of African descent (c.2988+1G>A). Varying the spacer region length allowed placement of the editing window in a more efficient context and enabled use of alternate protospacer adjacent motifs. Using these modifications, we restored clinically significant levels of CFTR function to human airway epithelial cells from two donors bearing the c.2988+1G>A variant.

Navigating nutrition as a childhood cancer survivor: Understanding patient and family needs for nutrition interventions or education.

AIM: Nutrition challenges are common during childhood cancer treatment and can persist into survivorship, increasing the risk of non-communicable diseases. Evidence-based practice and implementation of nutrition interventions/education for childhood cancer survivors has been poorly investigated and may influence their future health. This study aimed to explore the nutrition interventions/education needs of childhood cancer survivors and the barriers and facilitators to delivering follow-up services in New Zealand. METHODS: Semi structured interviews were conducted with childhood cancer survivors and/or their families (n=22) and health professionals (n=9) from a specialist paediatric oncology centre in New Zealand. Interviews were audio-recorded and transcribed verbatim. Transcripts were analysed inductively using thematic analysis. A multi-level consensus coding methodology was used where each theme and associated subthemes were discussed with the study team for confirmation to ensure accurate coding and analysis. RESULTS: Three themes emerged from the analysis: (1) the current survivorship care pathway does not provide adequate interventions/education, (2) weight and dietary changes are common challenges and (3) requirements for interventions/education in survivorship are varied. Common nutrition-related concerns included fussy eating/limited dietary intake, poor diet quality, difficulties with tube weaning, and challenges with weight gain. Participants expressed a desire for education on healthy eating alongside information about cancer-related nutrition issues, such as learned food aversions. A preference for clear referral pathways and multifaceted interventions tailored to individual patient needs was identified. CONCLUSION: The trifecta of treatment side effects, negative feeding practices and poor messaging from health professionals creates a challenging environment to optimise nutrition. A stepped care model matching the intervention intensity with the childhood cancer survivors is required. Education for healthcare professionals will improve the delivery of timely interventions/education and monitoring practices.

Toward Gene Transfer Nanoparticles as Therapeutics.

Genetic medicine has great potential to treat the underlying causes of many human diseases with exquisite precision, but the field has historically been stymied by delivery as the central challenge. Nanoparticles, engineered constructs the size of natural viruses, are being designed to more closely mimic the delivery efficiency of viruses, while enabling the advantages of increased safety, cargo-carrying flexibility, specific targeting, and ease in manufacturing. The speed in which nonviral gene transfer nanoparticles are making progress in the clinic is accelerating, with clinical validation of multiple nonviral nucleic acid delivery nanoparticle formulations recently FDA approved for both expression and for silencing of genes. While much of this progress has been with lipid nanoparticle formulations, significant development is being made with other nanomaterials for gene transfer as well, with favorable attributes such as biodegradability, scalability, and cell targeting. This review highlights the state of the field, current challenges in delivery, and opportunities for engineered nanomaterials to meet these challenges, including enabling long-term therapeutic gene editing. Delivery technology utilizing different kinds of nanomaterials and varying cargos for gene transfer (DNA, mRNA, and ribonucleoproteins) are discussed. Clinical applications are presented, including for the treatment of genetic diseases such as cystic fibrosis.

Women as health care decision-makers: implications for health care coverage in the United States.

Women in the United States make approximately 80% of the health care decisions for their families, yet often go without health care coverage themselves. The implementation of the Affordable Care Act provides an historical opportunity for women to gain health care coverage for themselves and their families. The focus of this commentary is on women's leadership roles in the context of health care decision- making and Affordable Care Act education and outreach, and implications for reaching broader health and social goals.

Parent-daughter communication about abortion among nonpregnant African-American adolescent females.

PURPOSE: To identify correlates of parent-daughter communication about abortion among nonpregnant adolescents. METHODS: Nonpregnant African-American females aged 14-17 years attending one of three charter schools on Chicago's South Side were recruited and surveyed regarding parental relationship quality, attitudes toward sexual health, and communication about sexual health. We calculated odds ratios (ORs) for ever discussing abortion using multivariable logistic regression. RESULTS: Of the 265 adolescents, 43.0% had ever discussed abortion with a parent. While 72.3% would voluntarily tell a parent about an abortion plan, 19.2% feared a parent would physically hurt her, punish her, or evict her. Ever communicating about abortion was significantly associated with having a mother who had a teen pregnancy (OR, 1.99; 95% confidence interval [CI], 1.05-3.78); having a prochoice abortion attitude (OR, 3.27; 95% CI, 1.29-8.25); having discussed sexually transmitted infections (OR, 2.97; 95% CI, 1.13-7.77) or birth control (OR, 2.96; 95% CI, 1.35-6.47) with a parent; and perceived parental approval of adolescent sexual activity (OR, 3.07; 95% CI, 1.20-7.88). Ever communicating about abortion was inversely related to being sexually experienced (OR, .48; 95% CI, .23-.99) or anticipating supportive parental reactions to an abortion decision (OR, .31; 95% CI, .13-.75). CONCLUSIONS: Although almost half of our sample was sexually active, the majority had never talked to their parents about abortion. Some reported fears of harm should the situation arise. Public policy should focus on promoting parent-daughter communication before an unwanted pregnancy rather than forcing communication after it occurs.

Abortion-seeking minors' views on the Illinois parental notification law: a qualitative study.

CONTEXT: Thirty-seven states have laws in effect that mandate parental involvement in adolescent abortion decisions. Little is known about minors' opinions of parental involvement laws. METHODS: In-depth interviews were conducted with 30 minors presenting for an abortion at one of three Chicago-area clinics in 2010. Interviewers described the Illinois parental notification law (which was passed in 1995 but is not in effect because of legal challenges) and a corresponding judicial bypass option to the minors and asked their opinions about them. Interviews were coded and analyzed using content analysis and grounded theory methods. RESULTS: Most minors perceived the law negatively, citing fears that it would lead to diminished reproductive autonomy for minors, forced continuation of pregnancies, adverse parental reactions (including emotional or physical abuse) and damaged parental relationships. A few held positive or ambivalent opinions, concluding that notifying a trusted adult could provide an adolescent with needed support, but that parental involvement should not be mandated. Most participants held negative opinions of judicial bypass, describing it as overwhelming and logistically complicated, and worrying that some minors might go to extreme lengths to avoid the process. CONCLUSIONS: Many minors have deep concerns about the potential harm that could result from parental involvement laws. These opinions provide a valuable addition to the debate on such laws, which purportedly are intended to ensure minors' best interests.

Using daily text-message reminders to improve adherence with oral contraceptives: a randomized controlled trial.

OBJECTIVE: To estimate whether women receiving daily text-message reminders have increased oral contraceptive pill adherence compared with women not receiving reminders. METHODS: This randomized controlled trial estimated whether there was an effect of daily text-message reminders on oral contraceptive pill adherence of new oral contraceptive pill users. Pill-taking was tracked for 3 months by an electronic monitoring device with wireless data collection. During the study period, participants assigned the intervention received a daily reminder text message. Eighty-two women were assigned randomly to detect a 1.6+/-2.0 pill difference (90% power, 5% alpha, 15% dropout). RESULTS: Participants were 79% white, non-Hispanic, 99% high school graduates, and 99% nulliparous with a mean age of 22 years. Most reported condom use with past coital activity, and more than half reported prior emergency contraception use. The mean number of missed pills per cycle did not differ significantly between the groups: 4.9+/-3.0 for the text-message group and 4.6+/-3.5 for the control group (P=.60). The number of missed pills per cycle increased over the course of the study, but this pattern did not increase differentially between the groups. Adherence recorded by the electronic monitoring device indicated much poorer adherence than that recorded by patient diaries. Despite poor pill-taking, there were no pregnancies. CONCLUSION: Daily text-message reminders did not improve oral contraceptive pill adherence. Although the lack of benefit may be attributed to the frequent use of alternative reminder systems in the control group, the rate of missed pills when measured objectively was still very high in both groups. CLINICAL TRIAL REGISTRATION: Clinicaltrials.gov, www.clinicaltrials.gov, NCT00733707. LEVEL OF EVIDENCE: I.