FOXM1 promotes neurofibromatosis type 1-associated malignant peripheral nerve sheath tumor progression in a NUF2-dependent manner.

Malignant peripheral nerve sheath tumors (MPNSTs) are aggressive soft-tissue sarcomas characterized by poor prognosis and low drug response rates. Traditional chemo/radiotherapies show only mild benefits for patients with MPNSTs, and no targeted therapy is available in the clinic. A better understanding of the molecular background of MPNSTs is critical for the development of effective targeted therapies. Forkhead box M1 (FOXM1) has been implicated in the progression of many human malignancies, though its role in MPNSTs is unclear. In this study, using four Gene Expression Omnibus (GEO) datasets and a tissue microarray, we demonstrated that FOXM1 upregulation was associated with poor prognosis in patients with MPNSTs. FOXM1 overexpression and knockdown regulated the proliferation and colony formation of MPNST cells. Using bioinformatics analysis and luciferase reporter assays, we identified NUF2 as a direct downstream target of FOXM1. Both in vitro and in vivo experiments demonstrated that the induction of MPNST cell proliferation by FOXM1 was dependent on elevated NUF2 expression, as NUF2 knockdown abolished the FOXM1-induced proliferation of MPNST cells. Our study showed that the FOXM1-NUF2 axis mediates human MPNST progression and could be a potential therapeutic target.

RRM2 as a novel prognostic and therapeutic target of NF1-associated MPNST.

BACKGROUND: Malignant peripheral nerve sheath tumors (MPNSTs) are aggressive sarcomas that typically develop in the setting of neurofibromatosis type 1 (NF1) and cause significant morbidity. Conventional therapies are often ineffective for MPNSTs. Ribonucleotide reductase subunit M2 (RRM2) is involved in DNA synthesis and repair, and is overexpressed in multiple cancers. However, its role in NF1-associated MPNSTs remains unknown. Our objective was to determine the therapeutic and prognostic potential of RRM2 in NF1-associated MPNSTs. METHODS: Identification of hub genes was performed by using NF1-associated MPNST microarray datasets. We detected RRM2 expression by immunochemical staining in an MPNST tissue microarray, and assessed the clinical and prognostic significance of RRM2 in an MPNST cohort. RRM2 knockdown and the RRM2 inhibitor Triapine were used to assess cell proliferation and apoptosis in NF1-associated MPNST cells in vitro and in vivo. The underlying mechanism of RRM2 in NF1-associated MPNST was revealed by transcriptome analysis. RESULTS: RRM2 is a key hub gene and its expression is significantly elevated in NF1-associated MPNST. We revealed that high RRM2 expression accounted for a larger proportion of NF1-associated MPNSTs and confirmed the correlation of high RRM2 expression with poor overall survival. Knockdown of RRM2 inhibited NF1-associated MPNST cell proliferation and promoted apoptosis and S-phase arrest. The RRM2 inhibitor Triapine displayed dose-dependent inhibitory effects in vitro and induced significant tumor growth reduction in vivo in NF1-associated MPNST. Analysis of transcriptomic changes induced by RRM2 knockdown revealed suppression of the AKT-mTOR signaling pathway. Overexpression of RRM2 activates the AKT pathway to promote NF1-associated MPNST cell proliferation. CONCLUSIONS: RRM2 expression is significantly elevated in NF1-associated MPNST and that high RRM2 expression correlates with poorer outcomes. RRM2 acts as an integral part in the promotion of NF1-associated MPNST cell proliferation via the AKT-mTOR signaling pathway. Inhibition of RRM2 may be a promising therapeutic strategy for NF1-associated MPNST.

Establishment and characterization of an immortalized human giant congenital melanocytic nevi cell line.

Treatments for giant congenital melanocytic nevi (GCMN) are extremely limited. Thus, there is an urgent need for development of relevant targeted therapies. However, current lack of preclinical cell models restricts progress in GCMN research. In this study, we aimed to establish and characterize an immortalized GCMN cell line. GCMN cells were successfully immortalized by means of lentivirus-mediated simian virus 40 large T transfection. The immortalized GNC cell line (ImGNC) showed lower proliferation rate and higher melanin content than primary melanocytes. Expression levels of the differentiation gene MITF and stemness genes TWIST1, SNAI1, and FOXD3 were elevated in ImGNCs; however, the established ImGNC cell line was immortalized but not transformed. Sanger sequencing detected the heterozygous NRASQ61K mutation in ImGNCs, but not the BRAFV600E mutation. Despite carrying the NRASQ61K allele, ImGNCs demonstrated suppressed MAPK activation and elevated PI3K/Akt activation, as compared with primary melanocytes. Drug sensitivity analysis showed that ImGNCs are more sensitive to PI3K/Akt and Bcl-2 inhibitors than to MEK or ERK inhibitors. Unlike the proliferation-inhibiting effect of PI3K/Akt inhibitors, the Bcl-2 inhibitor navitoclax promptly promoted apoptosis in ImGNCs. Considering the low proliferation characteristics of GCMN in vivo, Bcl-2 may be a potential therapeutic target that warrants further research.

Combined Cyclin-Dependent Kinase Inhibition Overcomes MAPK/Extracellular Signal-Regulated Kinase Kinase Inhibitor Resistance in Plexiform Neurofibroma of Neurofibromatosis Type I.

MAPK/extracellular signal-regulated kinase kinase (MEK) 1/2 inhibitors (MEKis) have recently achieved surprising success in treating unresectable plexiform neurofibromas (PNFs). However, few studies have investigated the mechanisms of MEKi resistance in patients with PNF. We determined the efficacy of six different MEKis for treating PNFs, explored drug resistance mechanisms, and identified potential combination therapies to overcome resistance. By screening drug efficacy among six MEKis in human NF1-deficient PNF cell lines, TAK-733 was found to reduce PNF cell viability the most. We then cultured the TAK-733‒resistant cells and explored the potential targets for further treatment. Both high-throughput drug screening and RNA sequencing analyses of MEKi-resistant PNF cells identified cyclin-dependent kinase inhibitors as potential agents for PNFs. Dinaciclib, a cyclin-dependent kinase inhibitor, showed synergistic effects on MEKi-resistant cells. Coadministration of dinaciclib and TAK-733 significantly reduced cell viability and inhibited sphere formation and colony formation. Dinaciclib did not affect MEK signaling but decreased the expression of several prosurvival proteins, including survivin and cyclin-dependent kinase 1, to induce apoptosis and inhibit mitosis. TAK-733/dinaciclib combination therapy induced tumor reduction in PNF patient‒derived xenografts mouse models. Therefore, the combination of MEKi and cyclin-dependent kinase inhibitor may be promising for treating inoperable PNFs, especially when drug resistance exists. Our findings provide evidence for future clinical trials with MEKi-resistant patients with PNF.

Faculty Service-Learning Students as Home-Visitors: Outcomes of a Lifestyle Modification Program for Vulnerable Families With Residents in Rural Indonesian Communities.

Background: Risks attributed to chronic diseases, cancer, musculoskeletal discomfort, and infectious diseases among Indonesians were found to be associated with lifestyle behaviors, particularly in rural areas. The aim of this study was to examine the outcomes of a home-visiting lifestyle modification program on improving health risk behaviors among Indonesians living in rural areas. Methods: A total of 160 Indonesians living in rural hamlets in the Yogyakarta Region of Indonesia participated in the program in the period of June 21 to July 21, 2019. In the pre-intervention home interview, learning needs of diet, exercise, hand hygiene, and substance use were identified by using structured assessment tools. In the next home visit, the visitors provided health education and facilitated lifestyle planning based on the related affective and cognitive domains of learning. Subsequent follow-up interviews were conducted 3 weeks after intervention. Results: The results showed that the self-reported intake of vegetables, fruits, meat and salt, cooking with less oil, hand hygiene before eating, number of cigarettes smoked, and symptoms of muscle stiffness significantly improved after the intervention. The lifestyle modification program consisted of the affective and cognitive domains of learning, and could lead to the target behavioral changes in self-reported and observable measures over 1 month. Conclusions: The findings contributed to the framework of community-based health education for health risk reduction and behavioral modification in developing rural communities where health care resources were limited. Further studies with control groups and vigorous objective measures were recommended to elucidate its long-term impacts. The factors leading to its sustainability concerning collaborative care partnerships between community residents and faculty resources are worthy of continued exploration.

Selection of internal references for RT-qPCR assays in Neurofibromatosis type 1 (NF1) related Schwann cell lines.

Real-time quantitative PCR (RT-qPCR) has been widely applied in uncovering disease mechanisms and screening potential biomarkers. Internal reference gene selection determines the accuracy and reproducibility of data analyses. The aim of this study was to identify the optimal reference genes for the relative quantitative analysis of RT-qPCR in fourteen NF1 related cell lines, including non-tumor, benign and malignant Schwann cell lines. The expression characteristics of eleven candidate reference genes (RPS18, ACTB, B2M, GAPDH, PPIA, HPRT1, TBP, UBC, RPLP0, TFRC and RPL32) were screened and analyzed by four software programs: geNorm, NormFinder, BestKeeper and RefFinder. Results showed that GAPDH, the most frequently used internal reference gene, was significantly unstable between various cell lines. The combinational use of two reference genes (PPIA and TBP) was optimal in malignant Schwann cell lines and the use of single reference genes (PPIA or PRLP0) alone or in combination was optimal in benign Schwann cell lines. These recommended internal reference gene selections may improve the accuracy and reproducibility of RT-qPCR in gene expression analyses of NF1 related tumors.

Adaptation and psychometric testing of the hoarding rating scale (HRS): a self-administered screening scale for epidemiological study in Chinese population.

BACKGROUND: Hoarding disorder is a chronic and debilitating illness associated with restrictions on activities of daily living, compromised social and occupational functioning, and adverse health outcomes. However, researchers lack a brief and self-administered screening measurement to assess compulsive hoarding in the Chinese speaking population. This study aimed to adapt and validate the Hoarding Rating Scale-Interview (HRS-I) to as a tool for screening compulsive hoarding behavior in Chinese population. METHODS: This study comprised two phases. During Phase 1, the English-language HRS-I was translated into Chinese (CHRS) (comprehensible for most Chinese speaking population, e.g., Cantonese & Mandarin) and subjected to an equivalence check. In Phase 2, the CHRS was validated by examining internal consistency, stability, and construct validity. Different samples were used appropriately to verify the items and reflect the psychometric properties. RESULTS: In Phase 1, the CHRS yielded satisfactory content (S-CVI = 0.93) and face validity ratings (comprehensibility = 100%, N = 20 participants of general public with age 18-72) and the English and Chinese versions were found to be equivalent (ICC = 0.887; N = 60 university staff and students). Phase 2 revealed satisfactory levels of internal consistency (Cronbach's α = 0.86; corrected item-total correlation = 0.60-0.74; N = 820 participants of general public), 2-week test-retest reliability (ICC = 0.78; N = 60 university students), and construct validity (one-factor CFA solution matched with the hypothesized model, χ2/d.f. = 2.26, RMSEA = 0.049, CFI = 0.99, IFI = 0.99, NFI = 0.99; n = 520 participants of general public). CONCLUSIONS: This study provides sufficient evidence of the reliability and validity of the CHRS for compulsive hoarding behavior screening in the Chinese population through self-administered method.

Use of tracking technology to examine life-space mobility among people with depression: a systematic review protocol.

INTRODUCTION: People with depression often experience disabilities that limit their social and physical capacity, daily function, and quality of life. Depressive symptoms and their implications on daily activities are often measured retrospectively using subjective measurement tools. Recently, more objective and accurate electronic data collection methods have been used to describe the daily life of people with depressive disorders. The results, however, have not yet been systematically reviewed. We aim to provide a knowledge basis for the use of tracking technologies in examining life-space mobility among adults with depression and those with anxiety as a comorbidity. METHODS AND ANALYSIS: A systematic review with a narrative approach for different types of study design will be conducted. The following databases will be used to gather data from 1994 to the present: MEDLINE, Cumulative Index to Nursing and Allied Health Literature, PsycINFO, Embase, Cochrane Library, Scopus, Web of Science, Health Technology Assessment Database and IEEE Xplore. The study selection will follow the Preferred Reporting Items for Systematic Review and Meta-Analysis Protocols. Methodological appraisal of studies will be performed using the Crowe Critical Appraisal Tool as well as the Cochrane Risk-of-Bias Tool for randomised controlled trials. A narrative synthesis of all included studies will be conducted. ETHICS AND DISSEMINATION: Because there will be no human involvement in the actual systematic review, no ethical approval will be required. The results will be disseminated in a peer-reviewed journal and in a conference presentation. PROSPERO REGISTRATION NUMBER: CRD42019127102.