Combined CDK4/6 and ERK1/2 Inhibition Enhances Antitumor Activity in NF1-Associated Plexiform Neurofibroma.

PURPOSE: Plexiform neurofibromas (PNF) are peripheral nerve sheath tumors that cause significant morbidity in persons with neurofibromatosis type 1 (NF1), yet treatment options remain limited. To identify novel therapeutic targets for PNF, we applied an integrated multi-omic approach to quantitatively profile kinome enrichment in a mouse model that has predicted therapeutic responses in clinical trials for NF1-associated PNF with high fidelity. EXPERIMENTAL DESIGN: Utilizing RNA sequencing combined with chemical proteomic profiling of the functionally enriched kinome using multiplexed inhibitor beads coupled with mass spectrometry, we identified molecular signatures predictive of response to CDK4/6 and RAS/MAPK pathway inhibition in PNF. Informed by these results, we evaluated the efficacy of the CDK4/6 inhibitor, abemaciclib, and the ERK1/2 inhibitor, LY3214996, alone and in combination in reducing PNF tumor burden in Nf1flox/flox;PostnCre mice. RESULTS: Converging signatures of CDK4/6 and RAS/MAPK pathway activation were identified within the transcriptome and kinome that were conserved in both murine and human PNF. We observed robust additivity of the CDK4/6 inhibitor, abemaciclib, in combination with the ERK1/2 inhibitor, LY3214996, in murine and human NF1(Nf1) mutant Schwann cells. Consistent with these findings, the combination of abemaciclib (CDK4/6i) and LY3214996 (ERK1/2i) synergized to suppress molecular signatures of MAPK activation and exhibited enhanced antitumor activity in Nf1flox/flox;PostnCre mice in vivo. CONCLUSIONS: These findings provide rationale for the clinical translation of CDK4/6 inhibitors alone and in combination with therapies targeting the RAS/MAPK pathway for the treatment of PNF and other peripheral nerve sheath tumors in persons with NF1.

Attitudes toward Medication for Opioid Use Disorder among Substance Use Treatment Providers.

Background: Provider attitudes can be a powerful reinforcer of stigma toward medication for opioid use disorder (MOUD). This study examines attitudes toward MOUD among substance use treatment providers and identifies personal and professional characteristics associated with more positive attitudes. Methods: Treatment providers (N = 570) working at publicly-funded substance use programs in Michigan self-administered a web-based survey (November 2020 through July 2021), reporting their socio-demographics, professional experience, and attitudes toward MOUD. Linear regression was used to identify factors associated with general attitudes toward MOUD and three logistic regression models were calculated to identify factors associated with perceptions of each medication. Results: Half of providers considered methadone an effective treatment (53.0%); 62.9% considered buprenorphine effective, and 70.3% considered naltrexone effective. Receipt of training (B = 1.433, p = .009) and serving pregnant women or women with children (B = 1.662, p < .001) were associated with more positive attitudes toward MOUD. Providers with advanced degrees were more likely to consider methadone (OR = 2.264, p = .006), buprenorphine (OR = 2.192, p = .009), and naltrexone (OR = 2.310, p = .011) effective. Rural providers were more likely to consider naltrexone effective (OR = 2.708, p = .003). Providers working with criminal legal populations were more likely to consider buprenorphine (OR = 2.948, p = .041) and naltrexone (OR = 4.108, p = .010) effective, but not methadone. Conclusion: Treatment providers' attitudes remain poorly aligned with the evidence base. Increased efforts are needed to address attitudes toward MOUD among the specialized treatment workforce.

Sex-Dependent Synaptic Remodeling of the Somatosensory Cortex in Mice With Prenatal Methadone Exposure.

Rising opioid use among pregnant women has led to a growing population of neonates exposed to opioids during the prenatal period, but how opioids affect the developing brain remains to be fully understood. Animal models of prenatal opioid exposure have discovered deficits in somatosensory behavioral development that persist into adolescence suggesting opioid exposure induces long lasting neuroadaptations on somatosensory circuitry such as the primary somatosensory cortex (S1). Using a mouse model of prenatal methadone exposure (PME) that displays delays in somatosensory milestone development, we performed an un-biased multi-omics analysis and investigated synaptic functioning in the primary somatosensory cortex (S1), where touch and pain sensory inputs are received in the brain, of early adolescent PME offspring. PME was associated with numerous changes in protein and phosphopeptide abundances that differed considerably between sexes in the S1. Although prominent sex effects were discovered in the multi-omics assessment, functional enrichment analyses revealed the protein and phosphopeptide differences were associated with synapse-related cellular components and synaptic signaling-related biological processes, regardless of sex. Immunohistochemical analysis identified diminished GABAergic synapses in both layer 2/3 and 4 of PME offspring. These immunohistochemical and proteomic alterations were associated with functional consequences as layer 2/3 pyramidal neurons revealed reduced amplitudes and a lengthened decay constant of inhibitory postsynaptic currents. Lastly, in addition to reduced cortical thickness of the S1, cell-type marker analysis revealed reduced microglia density in the upper layer of the S1 that was primarily driven by PME females. Taken together, our studies show the lasting changes on synaptic function and microglia in S1 cortex caused by PME in a sex-dependent manner.

A multimodal analysis of physical activity, sleep, and work shift in nurses with wearable sensor data.

Night shift workers are often associated with circadian misalignment and physical discomfort, which may lead to burnout and decreased work performance. Moreover, the irregular work hours can lead to significant negative health outcomes such as poor eating habits, smoking, and being sedentary more often. This paper uses commercial wearable sensors to explore correlates and differences in the level of physical activity, sleep, and circadian misalignment indicators among day shift nurses and night shift nurses. We identify which self-reported assessments of affect, life satisfaction, and sleep quality, are associated with physiological and behavioral signals captured by wearable sensors. The results using data collected from 113 nurses in a large hospital setting, over a period of 10 weeks, indicate that night shift nurses are more sedentary, and report lower levels of life satisfaction than day-shift nurses. Moreover, night shift nurses report poorer sleep quality, which may be correlated with challenges in their attempts to fall asleep on off-days.

17ß-Estradiol and estrogen receptor α protect right ventricular function in pulmonary hypertension via BMPR2 and apelin.

Women with pulmonary arterial hypertension (PAH) exhibit better right ventricular (RV) function and survival than men; however, the underlying mechanisms are unknown. We hypothesized that 17ß-estradiol (E2), through estrogen receptor α (ER-α), attenuates PAH-induced RV failure (RVF) by upregulating the procontractile and prosurvival peptide apelin via a BMPR2-dependent mechanism. We found that ER-α and apelin expression were decreased in RV homogenates from patients with RVF and from rats with maladaptive (but not adaptive) RV remodeling. RV cardiomyocyte apelin abundance increased in vivo or in vitro after treatment with E2 or ER-α agonist. Studies employing ER-α-null or ER-ß-null mice, ER-α loss-of-function mutant rats, or siRNA demonstrated that ER-α is necessary for E2 to upregulate RV apelin. E2 and ER-α increased BMPR2 in pulmonary hypertension RVs and in isolated RV cardiomyocytes, associated with ER-α binding to the Bmpr2 promoter. BMPR2 is required for E2-mediated increases in apelin abundance, and both BMPR2 and apelin are necessary for E2 to exert RV-protective effects. E2 or ER-α agonist rescued monocrotaline pulmonary hypertension and restored RV apelin and BMPR2. We identified what we believe to be a novel cardioprotective E2/ER-α/BMPR2/apelin axis in the RV. Harnessing this axis may lead to novel RV-targeted therapies for PAH patients of either sex.

Transcriptomic modifications in developmental cardiopulmonary adaptations to chronic hypoxia using a murine model of simulated high-altitude exposure.

Mechanisms driving adaptive developmental responses to chronic high-altitude (HA) exposure are incompletely known. We developed a novel rat model mimicking the human condition of cardiopulmonary adaptation to HA starting at conception and spanning the in utero and postnatal timeframe. We assessed lung growth and cardiopulmonary structure and function and performed transcriptome analyses to identify mechanisms facilitating developmental adaptations to chronic hypoxia. To generate the model, breeding pairs of Sprague-Dawley rats were exposed to hypobaric hypoxia (equivalent to 9,000 ft elevation). Mating, pregnancy, and delivery occurred in hypoxic conditions. Six weeks postpartum, structural and functional data were collected in the offspring. RNA-Seq was performed on right ventricle (RV) and lung tissue. Age-matched breeding pairs and offspring under room air (RA) conditions served as controls. Hypoxic rats exhibited significantly lower body weights and higher hematocrit levels, alveolar volumes, pulmonary diffusion capacities, RV mass, and RV systolic pressure, as well as increased pulmonary artery remodeling. RNA-Seq analyses revealed multiple differentially expressed genes in lungs and RVs from hypoxic rats. Although there was considerable similarity between hypoxic lungs and RVs compared with RA controls, several upstream regulators unique to lung or RV were identified. We noted a pattern of immune downregulation and regulation patterns of immune and hormonal mediators similar to the genome from patients with pulmonary arterial hypertension. In summary, we developed a novel murine model of chronic hypoxia exposure that demonstrates functional and structural phenotypes similar to human adaptation. We identified transcriptomic alterations that suggest potential mechanisms for adaptation to chronic HA.

Understanding Public Attitudes Toward Cannabis Legalization: Qualitative Findings From a Statewide Survey.

BACKGROUND: Cannabis policy is rapidly evolving in the United States as more states legalize medical and non-medical marijuana. Public opinion has shifted dramatically in favor of marijuana legalization. OBJECTIVES: This study examines the reasons that people support, oppose, or are unsure about marijuana legalization, focusing on the participants' own words. METHODS: A statewide sample of adults (N = 2,608) in Michigan completed an online survey about marijuana legalization (August and September 2016). Participants indicated whether they supported, opposed, or were unsure about marijuana legalization, and were then prompted to complete an open-ended response explaining the main reasons for their view. Thematic analysis was then used to code the open-ended responses (n = 2,054) and analytic induction was used to evaluate the coding. RESULTS: 48.1% of the sample supported cannabis legalization, 41.9% were opposed to legalization, and 10% were unsure. Harms associated with marijuana use were the most commonly given reasons for opposing legalization. Those who supported legalization were most likely to state that marijuana is less dangerous than other substances and has medical benefits. They also cited criminal justice reform and the potential for tax revenue as potential benefits of legalization. Reasons for supporting and opposing legalization differed based on gender, age, and recent marijuana use. Conclusions/Importance: Findings highlight nuances in public attitudes toward cannabis legalization. Many who support cannabis legalization recognize some potential negative consequences of these policy changes. Understanding views of cannabis is important as policies for marijuana use and sales become less restrictive.