Nf1 deficiency modulates the stromal environment in the pretumorigenic rat mammary gland.

Background: Neurofibromin, coded by the NF1 tumor suppressor gene, is the main negative regulator of the RAS pathway and is frequently mutated in various cancers. Women with Neurofibromatosis Type I (NF1)-a tumor predisposition syndrome caused by a germline NF1 mutation-have an increased risk of developing aggressive breast cancer with poorer prognosis. The mechanism by which NF1 mutations lead to breast cancer tumorigenesis is not well understood. Therefore, the objective of this work was to identify stromal alterations before tumor formation that result in the increased risk and poorer outcome seen among NF1 patients with breast cancer. Approach: To accurately model the germline monoallelic NF1 mutations in NF1 patients, we utilized an Nf1-deficient rat model with accelerated mammary development before presenting with highly penetrant breast cancer. Results: We identified increased collagen content in Nf1-deficient rat mammary glands before tumor formation that correlated with age of tumor onset. Additionally, gene expression analysis revealed that Nf1-deficient mature adipocytes in the rat mammary gland have increased collagen expression and shifted to a fibroblast and preadipocyte expression profile. This alteration in lineage commitment was also observed with in vitro differentiation, however, flow cytometry analysis did not show a change in mammary adipose-derived mesenchymal stem cell abundance. Conclusion: Collectively, this study uncovered the previously undescribed role of Nf1 in mammary collagen deposition and regulating adipocyte differentiation. In addition to unraveling the mechanism of tumor formation, further investigation of adipocytes and collagen modifications in preneoplastic mammary glands will create a foundation for developing early detection strategies of breast cancer among NF1 patients.

Screening for serious mental illness in a correctional setting.

BACKGROUND: Serious mental illness (SMI) is more common among adults in correctional settings than in the general population. No standard exists for SMI screening across correctional settings; SMI therefore often goes undetected in these facilities. Placing individuals with unidentified SMI who are incarcerated in general population cells increases their risk for self-harming behaviors, suicide, and for being victims or perpetrators of exploitation and violence. METHODS: This article describes a quantitative, descriptive study conducted to evaluate the use of the Brief Jail Mental Health Screen (BJMHS) tool to screen for possible SMI among individuals in a jail setting. RESULTS: A total of 89 individuals who were incarcerated in one jail setting were screened either with the facility's internally developed standard medical questionnaire (SMQ) or with the BJMHS. Findings showed that 28% screened positive for possible SMI using the BJMHS as compared with only 3% using the SMQ. CONCLUSION: The BJMHS flagged a higher number of possible instances of SMI than the jail's SMQ, potentially signifying its screening superiority. Identification of SMI leads to better care for individuals who are incarcerated, and it increases safety for the individual with SMI, the greater jail population, and jail staff. Findings from this study were shared with system leadership, which has replaced other screening tools with the BJMHS in at least 250 correctional facilities throughout the US.

p53 modulates kinase inhibitor resistance and lineage plasticity in NF1-related MPNSTs.

Malignant peripheral nerve sheath tumors (MPNSTs) are chemotherapy resistant sarcomas that are a leading cause of death in neurofibromatosis type 1 (NF1). Although NF1-related MPNSTs derive from neural crest cell origin, they also exhibit intratumoral heterogeneity. TP53 mutations are associated with significantly decreased survival in MPNSTs, however the mechanisms underlying TP53-mediated therapy responses are unclear in the context of NF1-deficiency. We evaluated the role of two commonly altered genes, MET and TP53, in kinome reprograming and cellular differentiation in preclinical MPNST mouse models. We previously showed that MET amplification occurs early in human MPNST progression and that Trp53 loss abrogated MET-addiction resulting in MET inhibitor resistance. Here we demonstrate a novel mechanism of therapy resistance whereby p53 alters MET stability, localization, and downstream signaling leading to kinome reprogramming and lineage plasticity. Trp53 loss also resulted in a shift from RAS/ERK to AKT signaling and enhanced sensitivity to MEK and mTOR inhibition. In response to MET, MEK and mTOR inhibition, we observed broad and heterogeneous activation of key differentiation genes in Trp53-deficient lines suggesting Trp53 loss also impacts lineage plasticity in MPNSTs. These results demonstrate the mechanisms by which p53 loss alters MET dependency and therapy resistance in MPNSTS through kinome reprogramming and phenotypic flexibility.

The DNP project: Two decades of impact.

ABSTRACT: Twenty years ago, pioneer Doctor of Nursing Practice (DNP) program students completed the first DNP projects. Today, DNP projects remain a requirement for graduation. This article illustrates how the DNP project can be integrated throughout the DNP curriculum to maximize effectiveness and promote the introduction, reinforcement, and evaluation of competencies in the new American Association of Colleges of Nursing Essentials.

NF1 deficiency drives metabolic reprogramming in ER+ breast cancer.

OBJECTIVE: NF1 is a tumor suppressor gene and its protein product, neurofibromin, is a negative regulator of the RAS pathway. NF1 is one of the top driver mutations in sporadic breast cancer such that 27 % of breast cancers exhibit damaging NF1 alterations. NF1 loss-of-function is a frequent event in the genomic evolution of estrogen receptor (ER)+ breast cancer metastasis and endocrine resistance. Individuals with Neurofibromatosis type 1 (NF) - a disorder caused by germline NF1 mutations - have an increased risk of dying from breast cancer [1-4]. NF-related breast cancers are associated with decreased overall survival compared to sporadic breast cancer. Despite numerous studies interrogating the role of RAS mutations in tumor metabolism, no study has comprehensively profiled the NF1-deficient breast cancer metabolome to define patterns of energetic and metabolic reprogramming. The goals of this investigation were (1) to define the role of NF1 deficiency in estrogen receptor-positive (ER+) breast cancer metabolic reprogramming and (2) to identify potential targeted pathway and metabolic inhibitor combination therapies for NF1-deficient ER + breast cancer. METHODS: We employed two ER+ NF1-deficient breast cancer models: (1) an NF1-deficient MCF7 breast cancer cell line to model sporadic breast cancer, and (2) three distinct, Nf1-deficient rat models to model NF-related breast cancer [1]. IncuCyte proliferation analysis was used to measure the effect of NF1 deficiency on cell proliferation and drug response. Protein quantity was assessed by Western Blot analysis. We then used RNAseq to investigate the transcriptional effect of NF1 deficiency on global and metabolism-related transcription. We measured cellular energetics using Agilent Seahorse XF-96 Glyco Stress Test and Mito Stress Test assays. We performed stable isotope labeling and measured [U-13C]-glucose and [U-13C]-glutamine metabolite incorporation and measured total metabolite pools using mass spectrometry. Lastly, we used a Bliss synergy model to investigate NF1-driven changes in targeted and metabolic inhibitor synergy. RESULTS: Our results revealed that NF1 deficiency enhanced cell proliferation, altered neurofibromin expression, and increased RAS and PI3K/AKT pathway signaling while constraining oxidative ATP production and restricting energetic flexibility. Neurofibromin deficiency also increased glutamine influx into TCA intermediates and dramatically increased lipid pools, especially triglycerides (TG). Lastly, NF1 deficiency alters the synergy between metabolic inhibitors and traditional targeted inhibitors. This includes increased synergy with inhibitors targeting glycolysis, glutamine metabolism, mitochondrial fatty acid transport, and TG synthesis. CONCLUSIONS: NF1 deficiency drives metabolic reprogramming in ER+ breast cancer. This reprogramming is characterized by oxidative ATP constraints, glutamine TCA influx, and lipid pool expansion, and these metabolic changes introduce novel metabolic-to-targeted inhibitor synergies.

Using unfolding case scenarios to promote clinical reasoning for nurse practitioner students.

ABSTRACT: This article describes how an unfolding case study can be used to promote the development of clinical reasoning through students' self-reported perceptions, although at the same time facilitating collaboration among providers from various specialties. An unfolding case (evolving case) provides sequential information about a patient's illness trajectory as they experience the illness and related symptomology. An unfolding case study was implemented during a college skills laboratory immersion experience for 33 nurse practitioner (NP) students who were in their final year of the NP program. Students were invited per email to complete a confidential REDCap survey after the case presentation and discussion. Twenty-three students completed the survey. More than half of the students (52%) stated the review of the unfolding case offered "significant learning value" and 78% rated the unfolding case as being "very to extremely" helpful in creating opportunities for critical thinking and engagement in clinical reasoning. Implementing unfolding case studies in NP student program curricula promotes critical thinking, clinical reasoning, and allows opportunities to engage in interprofessional collaboration.

An Integrative Review of the Barriers and Facilitators to Nurse Engagement in Quality Improvement in the Clinical Practice Setting.

BACKGROUND: Nurse engagement in quality improvement (QI) improves health care quality and outcomes but is typically low in clinical settings. PURPOSE: An integrative review was conducted to identify facilitators and barriers of nurse engagement in QI. METHODS: This integrative review was conducted using an electronic search of databases with search terms specific to nursing engagement in QI. The Johns Hopkins Nursing Evidence-Based Practice Evidence Level and Quality Guide was used to rate quality and level of evidence. RESULTS: Nine articles met the criteria for review. Top barriers were leadership, education and training, resource constraints, data, culture, and time. Top facilitators were leadership, education and training, culture, mentors, and champions. CONCLUSION: High-quality literature exploring barriers and facilitators of nurse engagement in QI is lacking. Research is needed to examine the degree to which these barriers and facilitators impact engagement and how they can be addressed to increase it.

Adapting clinical experiences during a pandemic: One college of nursing's approach.

ABSTRACT: Graduate level nursing programs are having increasing difficulty obtaining clinical sites for their students. Nurse practitioner (NP) students need a minimum of 500 direct care hours in addition to indirect hours. Simulation experiences may not be used as direct patient care hours for NP students, but telehealth experiences may be used if the focus is on obtaining similar competencies to what would be acquired with face-to-face patient experiences. One college of nursing adapted several opportunities for NP students to acquire indirect clinical experiences and specific NP competencies during the coronavirus (COVID-19) pandemic. Several different clinically focused teaching methodologies were planned and implemented. These varied by NP specialty track and included complex case analyses, virtual (Zoom) rounds, well-child cases, and objective structured clinical examinations. These adapted clinical experiences are effective methods of helping students acquire clinical competencies and skills; however, they bring their own challenges. Post-COVID may be a time of new beginnings, moving nursing education forward on finding new and better ways to ensure that students acquire clinical competencies. What the new normal will be for nursing education is yet to be determined, but nursing education will likely never return to where we were before the pandemic.

Strengthening the impact of Doctor of Nursing Practice projects in education and clinical practice.

ABSTRACT: The significance and value of Doctor of Nursing Practice (DNP) projects can be potentiated by collaboration between educators and practice leaders and when students build on previous DNP projects, as demonstrated in this article. Projects can have a longstanding financial, quality, and outcome impact, helping demonstrate the worth and efficacy of the DNP degree.

Quality improvement engagement and competence: A comparison between frontline nurses and nurse leaders.

BACKGROUND: Nurses play a pivotal role in improving patient care. To maximize nurses' impact on quality, nurses must have quality improvement (QI) competence and engage fully in QI initiatives. PURPOSE: To describe QI competence (knowledge, skills, and attitudes) among frontline nurses and leaders; and compare variations in competence among nursing roles, experience, and specialty areas. METHODS: A total of 681 nurses at one heath system fully completed the Nursing Quality Improvement Practice tool electronically. FINDINGS: Half of the respondents reported QI engagement (53.6%). Mean knowledge scores were 5.08 (SD 1.16, 7 items). Skill proficiency was low (M = 2.82, SD = 1.03; range 1-6) although QI attitudes were favorable (M = 3.76, SD = 0.63; range 1-5). Significant differences in skills and attitudes were identified by role. QI competence among nurses employed in various specialty areas were similar. DISCUSSION: Strategies for increasing QI competence and engagement of nurses must be created and deployed in order to improve quality and safety.

Development of the Nursing Quality Improvement in Practice Tool: Advancing Frontline Nursing Practice.

BACKGROUND: Frontline nurse engagement in quality improvement (QI) improves nurse-sensitive outcomes; yet research suggests frontline nurses are not engaging in QI. PURPOSE: The purpose of this study was to develop, refine, and psychometrically evaluate the Nursing Quality Improvement in Practice (N-QuIP) tool to measure nurses' competency, engagement, and barriers/facilitators to QI engagement. METHODS: Item development was guided by an expert panel and literature review. Factor analysis and reliability indices were assessed through 681 surveys completed by nurses at one medical center. RESULTS: Cronbach α coefficients were 0.97 (Skill Scale) and 0.90 (Attitude Scale). Kuder-Richardson Formula 20 (KR-20) for knowledge was 0.36. Exploratory factor analysis identified 4 (Skill) and 3 (Attitude) subscales respectively, aligning well with QI competencies. CONCLUSIONS: Preliminary data suggest that the N-QuIP is a valid and reliable tool for assessing nurse QI competence and engagement. Understanding current knowledge, skills, and attitudes and identified barriers/facilitators can help the development of strategies aimed at increasing QI engagement.

Dissecting the Rat Mammary Gland: Isolation, Characterization, and Culture of Purified Mammary Epithelial Cells and Fibroblasts.

With the advent of CRISPR-Cas and the ability to easily modify the genome of diverse organisms, rat models are being increasingly developed to interrogate the genetic events underlying mammary development and tumorigenesis. Protocols for the isolation and characterization of mammary epithelial cell subpopulations have been thoroughly developed for mouse and human tissues, yet there is an increasing need for rat-specific protocols. To date, there are no standard protocols for isolating rat mammary epithelial subpopulations. Analyzing changes in the rat mammary hierarchy will help us elucidate the molecular events in breast cancer, the cells of origin for breast cancer subtypes, and the impact of the tumor microenvironment. Here we describe several methods developed for 1) rat mammary epithelial cell isolation; 2) rat mammary fibroblast isolation; 3) culturing rat mammary epithelial cells; and characterization of rat mammary cells by 4) flow cytometric analysis; and 5) immunofluorescence. Cells derived from this protocol can be used for many purposes, including RNAseq, drug studies, functional assays, gene/protein expression analyses, and image analysis.

DNP Program Faculty and Graduates' Knowledge and Use of QI and Safety Processes.

BACKGROUND: Across doctor of nursing practice (DNP) programs, wide variation exists for DNP project expectations and curricular integration of Quality and Safety Education for Nurses competencies. PURPOSE: This study examined DNP project characteristics and compared knowledge, skills, and attitudes about quality improvement (QI) processes between DNP program faculty and graduates. METHODS: This descriptive study used a national convenience sample of DNP program faculty and graduates from multiple settings (N = 147) who completed an electronic survey of a modified version of the Quality Improvement Knowledge, Skills, and Attitudes Survey. RESULTS: Most DNP projects were QI/practice improvement (85.5%) and required interprofessional collaboration (65%) and implementation of a practice change (55.8%) with evaluation (65.5%). DNP program faculty (n = 73) and graduates (n = 30) had no significant differences in QI knowledge or skills; faculty reported less confidence in their knowledge (P = .002) and skills (P = .007) than graduates. CONCLUSIONS: Faculty development efforts to improve QI knowledge and skills for DNP program faculty are needed.

NF1 deficiency correlates with estrogen receptor signaling and diminished survival in breast cancer.

The key negative regulatory gene of the RAS pathway, NF1, is mutated or deleted in numerous cancer types and is associated with increased cancer risk and drug resistance. Even though women with neurofibromatosis (germline NF1 mutations) have a substantially increased breast cancer risk at a young age and NF1 is commonly mutated in sporadic breast cancers, we have a limited understanding of the role of NF1 in breast cancer. We utilized CRISPR-Cas9 gene editing to create Nf1 rat models to evaluate the effect of Nf1 deficiency on tumorigenesis. The resulting Nf1 indels induced highly penetrant, aggressive mammary adenocarcinomas that express estrogen receptor (ER) and progesterone receptor (PR). We identified distinct Nf1 mRNA and protein isoforms that were altered during tumorigenesis. To evaluate NF1 in human breast cancer, we analyzed genomic changes in a data set of 2000 clinically annotated breast cancers. We found NF1 shallow deletions in 25% of sporadic breast cancers, which correlated with poor clinical outcome. To identify biological networks impacted by NF1 deficiency, we constructed gene co-expression networks using weighted gene correlation network analysis (WGCNA) and identified a network connected to ESR1 (estrogen receptor). Moreover, NF1-deficient cancers correlated with established RAS activation signatures. Estrogen-dependence was verified by estrogen-ablation in Nf1 rats where rapid tumor regression was observed. Additionally, Nf1 deficiency correlated with increased estrogen receptor phosphorylation in mammary adenocarcinomas. These results demonstrate a significant role for NF1 in both NF1-related breast cancer and sporadic breast cancer, and highlight a potential functional link between neurofibromin and the estrogen receptor.

Genomic Status of MET Potentiates Sensitivity to MET and MEK Inhibition in NF1-Related Malignant Peripheral Nerve Sheath Tumors.

Malignant peripheral nerve sheath tumors (MPNST) are highly resistant sarcomas that occur in up to 13% of individuals with neurofibromatosis type I (NF1). Genomic analysis of longitudinally collected tumor samples in a case of MPNST disease progression revealed early hemizygous microdeletions in NF1 and TP53, with progressive amplifications of MET, HGF, and EGFR To examine the role of MET in MPNST progression, we developed mice with enhanced MET expression and Nf1 ablation (Nf1fl/ko;lox-stop-loxMETtg/+;Plp-creERTtg/+ ; referred to as NF1-MET). NF1-MET mice express a robust MPNST phenotype in the absence of additional mutations. A comparison of NF1-MET MPNSTs with MPNSTs derived from Nf1ko/+;p53R172H;Plp-creERTtg/+ (NF1-P53) and Nf1ko/+;Plp-creERTtg/+ (NF1) mice revealed unique Met, Ras, and PI3K signaling patterns. NF1-MET MPNSTs were uniformly sensitive to the highly selective MET inhibitor, capmatinib, whereas a heterogeneous response to MET inhibition was observed in NF1-P53 and NF1 MPNSTs. Combination therapy of capmatinib and the MEK inhibitor trametinib resulted in reduced response variability, enhanced suppression of tumor growth, and suppressed RAS/ERK and PI3K/AKT signaling. These results highlight the influence of concurrent genomic alterations on RAS effector signaling and therapy response to tyrosine kinase inhibitors. Moreover, these findings expand our current understanding of the role of MET signaling in MPNST progression and identify a potential therapeutic niche for NF1-related MPNSTs.Significance: Longitudinal genomic analysis reveals a positive selection for MET and HGF copy number gain early in malignant peripheral nerve sheath tumor progression. Cancer Res; 78(13); 3672-87. ©2018 AACR.

Metabolic profiling of triple-negative breast cancer cells reveals metabolic vulnerabilities.

BACKGROUND: Among breast cancers, the triple-negative breast cancer (TNBC) subtype has the worst prognosis with no approved targeted therapies and only standard chemotherapy as the backbone of systemic therapy. Unique metabolic changes in cancer progression provide innovative therapeutic opportunities. The receptor tyrosine kinases (RTKs) epidermal growth factor receptor (EGFR), and MET receptor are highly expressed in TNBC, making both promising therapeutic targets. RTK signaling profoundly alters cellular metabolism by increasing glucose consumption and subsequently diverting glucose carbon sources into metabolic pathways necessary to support the tumorigenesis. Therefore, detailed metabolic profiles of TNBC subtypes and their response to tyrosine kinase inhibitors may identify therapeutic sensitivities. METHODS: We quantified the metabolic profiles of TNBC cell lines representing multiple TNBC subtypes using gas chromatography mass spectrometry. In addition, we subjected MDA-MB-231, MDA-MB-468, Hs578T, and HCC70 cell lines to metabolic flux analysis of basal and maximal glycolytic and mitochondrial oxidative rates. Metabolic pool size and flux measurements were performed in the presence and absence of the MET inhibitor, INC280/capmatinib, and the EGFR inhibitor, erlotinib. Further, the sensitivities of these cells to modulators of core metabolic pathways were determined. In addition, we annotated a rate-limiting metabolic enzymes library and performed a siRNA screen in combination with MET or EGFR inhibitors to validate synergistic effects. RESULTS: TNBC cell line models displayed significant metabolic heterogeneity with respect to basal and maximal metabolic rates and responses to RTK and metabolic pathway inhibitors. Comprehensive systems biology analysis of metabolic perturbations, combined siRNA and tyrosine kinase inhibitor screens identified a core set of TCA cycle and fatty acid pathways whose perturbation sensitizes TNBC cells to small molecule targeting of receptor tyrosine kinases. CONCLUSIONS: Similar to the genomic heterogeneity observed in TNBC, our results reveal metabolic heterogeneity among TNBC subtypes and demonstrate that understanding metabolic profiles and drug responses may prove valuable in targeting TNBC subtypes and identifying therapeutic susceptibilities in TNBC patients. Perturbation of metabolic pathways sensitizes TNBC to inhibition of receptor tyrosine kinases. Such metabolic vulnerabilities offer promise for effective therapeutic targeting for TNBC patients.

Glesatinib Exhibits Antitumor Activity in Lung Cancer Models and Patients Harboring MET Exon 14 Mutations and Overcomes Mutation-mediated Resistance to Type I MET Inhibitors in Nonclinical Models.

Purpose:MET exon 14 deletion (METex14 del) mutations represent a novel class of non-small cell lung cancer (NSCLC) driver mutations. We evaluated glesatinib, a spectrum-selective MET inhibitor exhibiting a type II binding mode, in METex14 del-positive nonclinical models and NSCLC patients and assessed its ability to overcome resistance to type I MET inhibitors.Experimental Design: As most MET inhibitors in clinical development bind the active site with a type I binding mode, we investigated mechanisms of acquired resistance to each MET inhibitor class utilizing in vitro and in vivo models and in glesatinib clinical trials.Results: Glesatinib inhibited MET signaling, demonstrated marked regression of METex14 del-driven patient-derived xenografts, and demonstrated a durable RECIST partial response in a METex14 del mutation-positive patient enrolled on a glesatinib clinical trial. Prolonged treatment of nonclinical models with selected MET inhibitors resulted in differences in resistance kinetics and mutations within the MET activation loop (i.e., D1228N, Y1230C/H) that conferred resistance to type I MET inhibitors, but remained sensitive to glesatinib. In vivo models exhibiting METex14 del/A-loop double mutations and resistance to type I inhibitors exhibited a marked response to glesatinib. Finally, a METex14 del mutation-positive NSCLC patient who responded to crizotinib but later relapsed, demonstrated a mixed response to glesatinib including reduction in size of a MET Y1230H mutation-positive liver metastasis and concurrent loss of detection of this mutation in plasma DNA.Conclusions: Together, these data demonstrate that glesatinib exhibits a distinct mechanism of target inhibition and can overcome resistance to type I MET inhibitors. Clin Cancer Res; 23(21); 6661-72. ©2017 AACR.

Quality and Safety Education for Nurses Competencies in Doctor of Nursing Practice Education: Exemplars From Education and Practice.

Graduate level Quality and Safety Education for Nurses (QSEN) competencies were created to facilitate preparation of the Doctor of Nursing Practice (DNP) graduate to have an impact on health care system changes, advance nursing practice, and improve the health of populations. Although there is agreement that QSEN competencies are important to DNP education, little is known about implementation of the competencies. This article describes the importance of QSEN competencies in DNP education and provides exemplars of QSEN competencies in DNP education and practice.

MET in human cancer: germline and somatic mutations.

Since the initial discovery of missense MET mutations in hereditary papillary renal carcinoma (HPRC), activating MET mutations have been identified in a diverse range of human cancers. MET mutations have been identified in several functional domains including the kinase, juxtamembrane, and Sema domains. Studies of these mutations have been invaluable for our understanding of the tumor initiating activity of MET, receptor tyrosine kinase (RTK) recycling and regulation, and mechanisms of resistance to kinase inhibition. These studies also demonstrate that mutationally activated MET plays a significant role in a wide range of cancers and RTKs can promote tumor progression through diverse mechanisms. This review will cover the various MET mutations that have been identified, their mechanism of action, and the significant role that mutationally-activated MET plays in tumor initiation, progression, and therapeutic resistance.

In vivo Efficacy Studies in Cell Line and Patient-derived Xenograft Mouse Models.

In vivo xenograft models derived from human cancer cells have been a gold standard for evaluating the genetic drivers of cancer and are valuable preclinical models for evaluating the efficacy of cancer therapeutics. Recently, patient-derived tumorgrafts from multiple tumor types have been developed and shown to more accurately recapitulate the molecular and histological heterogeneity of cancer. Here we detail the procedures for developing patient-derived xenograft models from breast cancer tissue, cell-based xenograft models, serial tumor transplantation, tumor measurement, and drug treatment.