Development of a Novel Assay for Direct Assessment of Selective Amylin Receptor Activation Reveals Novel Differences in Behavior of Selective and Nonselective Peptide Agonists.

Dual amylin and calcitonin receptor agonists (DACRAs) show promise as efficacious therapeutics for treatment of metabolic disease, including obesity. However, differences in efficacy in vivo have been observed for individual DACRAs, indicating that detailed understanding of the pharmacology of these agents across target receptors is required for rational drug development. To date, such understanding has been hampered by lack of direct, subtype-selective, functional assays for the amylin receptors (AMYRs). Here, we describe the generation of receptor-specific assays for recruitment of Venus-tagged Gs protein through fusion of luciferase to either the human calcitonin receptor (CTR), human receptor activity-modifying protein (RAMP)-1, RAMP1 (AMY1R), human RAMP2 (AMY2R), or human RAMP3 (AMY3R). These assays revealed a complex pattern of receptor activation by calcitonin, amylin, or DACRA peptides that was distinct at each receptor subtype. Of particular note, although both of the CT-based DACRAs, sCT and AM1784, displayed relatively similar behaviors at CTR and AMY1R, they generated distinct responses at AMY2R and AMY3R. These data aid the rationalization of in vivo differences in response to DACRA peptides in rodent models of obesity. Direct assessment of the pharmacology of novel DACRAs at AMYR subtypes is likely to be important for development of optimized therapeutics for treatment of metabolic diseases. SIGNIFICANCE STATEMENT: Amylin receptors (AMYRs) are important obesity targets. Here we describe a novel assay that allows selective functional assessment of individual amylin receptor subtypes that provides unique insight into the pharmacology of potential therapeutic ligands. Direct assessment of the pharmacology of novel agonists at AMYR subtypes is likely to be important for development of optimized therapeutics for treatment of metabolic diseases.

miR-130a expression is related to aortic dilation in bicuspid aortic valve children.

BACKGROUND: Bicuspid aortic valve disease (BAV) is present in 0.5-2% of the population and can promote aortic dilation, eventually leading to fatal consequences. Although some biomarkers have been proposed in adults, no studies have tested these candidates in children. We aimed to evaluate four miRNAs previously described to be related to BAV disease and aortic dilation in adults in a paediatric cohort. METHODS: Eighty participants ≤17 years old (4-17; mean 12) were included. From the BAV group, 40% had a dilated aorta (z score >2). RT‒qPCR were performed in plasma samples to quantify miR-122, miR-130a, miR-486, and miR-718 using the delta-delta Ct method. Functional and enrichment analyses of miR-130a were also performed. RESULTS: miR-130a expression in plasma was found to be significantly lower in BAV patients with a dilated aorta versus nondilated patients (p = 0.008) and healthy TAV controls (p = 0.004). Furthermore, miR-130a expression in plasma was inversely correlated with ascending aorta (r = 0.318, p = 0.004) and aortic root z scores (r = 0.322; p = 0.004). Enrichment analysis showed that miR-130a target genes are related to the TGFß signalling pathway. CONCLUSIONS: miR-130a expression in plasma is decreased in aortic-dilated BAV children compared to nondilated BAV children, helping differentiate low- to high-risk patients. IMPACT: miR-130a expression in plasma is related to aortic dilation in bicuspid aortic valve (BAV) children. To our knowledge, this is the first study that analyses miRNA patterns in bicuspid aortic valve children with aortic dilation. miR-130a expression in plasma could be a biomarker in order to help differentiate low-to high-risk BAV children, which is vitally important for advanced care planning.

High-risk pregnancy and its relationship with the neurodevelopment and behavior of 2-year-old children.

High-risk pregnancies elevate maternal stress, impacting offspring neurodevelopment and behavior. This study, involving 112 participants, aimed to compare perceived stress, neurodevelopment, and behavior in high-risk and low-risk pregnancies. Two groups, high-risk and low-risk, were assessed during pregnancy for stress using hair cortisol and psychological analysis. At 24 months post-birth, their children's neurodevelopment and behavior were evaluated. Results revealed higher perceived stress and pregnancy-related concerns in high-risk pregnancies, contrasting with low-risk pregnancies. Offspring from high-risk pregnancies displayed elevated internalizing behavior scores, while low-risk pregnancies showed higher externalizing behavior scores. Additionally, women in low-risk pregnancies exhibited increased cortisol concentrations 24 months post-delivery. These findings underscore the necessity for early stress detection and prevention programs during pregnancy, particularly in high-risk cases, to enhance maternal and infant health.

Delving into the Variability of Supramolecular Affinity: Self-Ion Pairing as a Central Player in Aqueous Host-Guest Chemistry.

The determination of binding constants is a key matter in evaluating the strength of host-guest interactions. However, the profound impact of self-ion pairing on this parameter is often underrated in aqueous solution, leading in some cases to a misinterpretation of the true potential of supramolecular assemblies. In the present study, we aim to shed further light on this critical factor by exploring the concentration-dependent behavior of a multicharged pillararene in water. Our observations reveal an extraordinary 1-million-fold variability in the affinity of this macrocycle toward a given anion, showcasing the highly dynamic character of electrostatic interactions. We argue that these findings bring to the forefront the inherent determinism that underlies the estimation of affinity constants, a factor profoundly shaped by both the sensitivity of the instrumental technique in use and the intricacies of the experimental design itself. In terms of applications, these results may provide the opportunity to optimize the operational concentrations of multicharged hosts in different scenarios, aiming to achieve their maximum efficiency based on the intended application. Unlocking the potential of this hidden variability may pave the way for the creation of novel molecular materials with advanced functionalities.

The effect of tyrosine supplementation on whole-body endurance performance in physically active population: A systematic review and meta-analysis including GRADE qualification.

Although tyrosine supplementation is well recognized to improve cognitive function, its impact on endurance performance is debatable and needs to be clarified further. The purpose of this systematic review and meta-analysis was to evaluate the effects of tyrosine supplementation on whole-body endurance performance in physically active population. The search strategy follow the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA), using four databases (Cochrane Library, Web of Science, Scopus, PsycINFO, and PubMed) until 3 August 2023. The effect of tyrosine (experimental condition) was compared against placebo (control condition). The methodological quality of the included studies was evaluated using the Physiotherapy Evidence Database (PEDro) scale. The Grading of Recommendations Assessment, Development, and Evaluation (GRADE Pro software) System was also used to assess the quality of evidence. A total of 10 interventions from 8 studies were included. The sub-group analysis revealed no significant differences between tyrosine and placebo conditions for time to exhaustion (SMD = 0.02; p = 0.94) and time trial performance (SMD = -0.04; p = 0.85). The level of evidence as qualified with GRADE was moderate. In conclusion, moderate-quality evidence suggests that tyrosine supplementation is ineffective on endurance performance in the physically active population, independently of the endurance task (TTE or ETT).

The role of resilience in the potential benefits of cognitive-behavioural stress management therapy during pregnancy.

PURPOSE: Stress during pregnancy has many negative repercussions on maternal and foetal health. It is therefore important to understand which therapies are effective in reducing stress levels and which variables influence the outcomes of these therapies. In this line, psychological resilience could play a key role. Thus, the aim of the study was to check whether pregnant women with different levels of resilience have different benefits in reducing cortisol levels, perceived stress, pregnancy worries, stress vulnerability, anxiety or depression through Cognitive Behavioural Stress Management Therapy. METHOD: The total sample consisted of 56 pregnant women: one group of pregnant women with high levels of resilience (n = 35); and another group with low levels of resilience (n = 21). Participants' cortisol concentration levels, perceived stress, pregnancy concerns, stress vulnerability, anxiety and depressive symptoms were assessed before and after therapy. Linear mixed models were performed to compare the two groups, which showed a group x time interaction for perceived stress. RESULTS: The low resilience group showed a reduction in their perceived stress levels with a medium effect after the intervention compared to the high resilience group, but no reduction was found in this group. No differences were found between the two groups on the other variables. CONCLUSION: Knowing which variables have a differential effect on the results of psychological therapy would allow delimiting the groups that obtain greater benefits from the therapy. This may lead to more efficient implementation of effective intervention programmes.

Pregnant women's mental health during the COVID-19 pandemic according to the trimester of pregnancy.

AIM: This study aimed to analyse the psychological impact of the COVID-19 pandemic on pregnant women according to the pregnancy trimester, comparing their psychopathological symptomatology, pregnancy-specific stress, resilience and perceived stress to those of women pregnant before the pandemic. METHODS: A total of 797 pregnant women participated in the study, one group of 393 women pregnant before the pandemic and the other of 404 women pregnant during the pandemic. Student-t test was used to analyse continuous data and the Chi-square test was used for categorical data. RESULTS: Psychopathological symptomatology was significantly higher in six subscales of the SCL-90-R in pregnant women during COVID-19: somatisation, interpersonal sensitivity, depression, anxiety, phobic anxiety, obsessions-compulsions, mainly on the first two trimesters. There is also a higher level of pregnancy-specific stress in pregnant women during the pandemic on the first two trimesters, most likely due to the hypervigilance and fears related to the COVID-19 disease. Nevertheless, perceived stress, usually elevated during pregnancy, was lower in women pregnant during the pandemic in comparison to those pregnant before, as a positive consequence of being on lockdown and diminishing the exposure to daily stressful situations. CONCLUSIONS: Knowing the struggles these women go through during each trimester of pregnancy can be the key to a better health professional-patient relationship, consequently having a positive impact on their mental and physical health.

Symptoms of Anxiety in Mothers During the COVID-19 Lockdown: A Structural Equation Model Approach.

The COVID-19 global pandemic has forced millions of people to stay confined at home, increasing symptoms of anxiety and stress levels. Women who are also mothers, for their part, not only face the demands of motherhood but must combine working life with family life locked down in their homes. Main objective was to develop an explanatory model of the psychological consequences of COVID-19 and parental and perceived stress in mothers. A total of 261 mothers were evaluated coinciding with the lockdown imposed by the Spanish Government. The model displayed adequate indices and it was found that symptoms of anxiety in mothers increased the levels of perceived stress. The model allows to understand the close relationships between the psychological consequences of lockdown and stress in mothers. Understanding these relationships will help to prepare and direct psychological interventions in this population in the case of a possible new surge.

Next-generation sequencing of bile cell-free DNA for the early detection of patients with malignant biliary strictures.

OBJECTIVE: Despite significant progresses in imaging and pathological evaluation, early differentiation between benign and malignant biliary strictures remains challenging. Endoscopic retrograde cholangiopancreatography (ERCP) is used to investigate biliary strictures, enabling the collection of bile. We tested the diagnostic potential of next-generation sequencing (NGS) mutational analysis of bile cell-free DNA (cfDNA). DESIGN: A prospective cohort of patients with suspicious biliary strictures (n=68) was studied. The performance of initial pathological diagnosis was compared with that of the mutational analysis of bile cfDNA collected at the time of first ERCP using an NGS panel open to clinical laboratory implementation, the Oncomine Pan-Cancer Cell-Free assay. RESULTS: An initial pathological diagnosis classified these strictures as of benign (n=26), indeterminate (n=9) or malignant (n=33) origin. Sensitivity and specificity of this diagnosis were 60% and 100%, respectively, as on follow-up 14 of the 26 and eight of the nine initially benign or indeterminate strictures resulted malignant. Sensitivity and specificity for malignancy of our NGS assay, herein named Bilemut, were 96.4% and 69.2%, respectively. Importantly, one of the four Bilemut false positives developed pancreatic cancer after extended follow-up. Remarkably, the sensitivity for malignancy of Bilemut was 100% in patients with an initial diagnosis of benign or indeterminate strictures. Analysis of 30 paired bile and tissue samples also demonstrated the superior performance of Bilemut. CONCLUSION: Implementation of Bilemut at the initial diagnostic stage for biliary strictures can significantly improve detection of malignancy, reduce delays in the clinical management of patients and assist in selecting patients for targeted therapies.

Spanish validation and factor structure of the Prenatal Distress Questionnaire Revised (NuPDQ).

BACKGROUND: Due to an absence of maternal stress being measured in routine prenatal care by clinicians, prenatal stress has become a serious problem which is associated with poorer obstetric outcomes, as well as worse maternal and infant health. For that reason, the aim of this study was the translation, validation and adaptation of Prenatal Distress Questionnaire Revised (NuPDQ) in a Spanish sample. METHODS: Three-hundred and seventy-one pregnant women were assessed using the NuPDQ, the Prenatal Distress Questionnaire, Perceived Stress Scale and the Connor-Davidson Resilience Scale. The NuPDQ was translated into Spanish by the backtranslation method and administered to participants. RESULTS: A confirmatory factor analysis revealed the established unidimensional structure to be a poor fit to data with the Spanish version. An exploratory factor analysis suggested a five-factor structure with 14 items. The instrument had good reliability, convergent and discriminant validity psychometric properties. CONCLUSIONS: The five-factor 14-item NuPDQ is useful to assess pregnancy-specific stress in Spanish pregnant women. It may be appropriate to use this instrument in order to identify pregnant women with high pregnancy-specific stress to try to prevent negative consequences derived from those high levels.

Resilience, stress and anxiety in pregnancy before and throughout the pandemic: a structural equation modelling approach.

The present study explored and compared the link between resilience and pregnancy-related stress, perceived stress, and anxiety, employing two structural equation models. One model focused on pregnant women before the outbreak of the pandemic, and the other on pregnancies throughout the pandemic. For this purpose, a total sample of 690 women during their pregnancy were collected: the Pre-Pandemic Group (P-PG) was composed of 341 pregnant women evaluated prior to the pandemic; and 349 pregnant women assessed at the time of the pandemic constituted the Pandemic Group (PG). The resilience, pregnancy-related stress, perceived stress, and anxiety symptomatology of the women were assessed. For both samples, resilience was found to lower levels of pregnancy-specific stress, as well as general perceived stress, and anxiety symptomatology. Furthermore, pregnancy-specific stress and perceived stress showed a covariance relationship and, that these, in turn, increased the anxiety. Moreover, the PG showed greater levels of pregnancy-specific stress, anxiety, somatisations, and obsessions-compulsions, while the P-PG presented higher perceived stress levels.

Usefulness of an immunohistochemical score in advanced pancreatic neuroendocrine tumors treated with CAPTEM or everolimus.

BACKGROUND: Pancreatic neuroendocrine tumors are rare neoplasms for which few predictive and/or prognostic biomarkers have been validated. Our previous work suggested the potential of the combined expression of N-myc downstream-regulated gen-1 (NDRG-1), O6-methylguanine DNA methyltransferase (MGMT) and Pleckstrin homology-like domain family A member 3 (PHLDA-3) as prognostic factors for relapse and survival. METHODS: In this new multicenter study we evaluated immunohistochemistry expression in 76 patients with advanced PanNET who were treated with capecitabine-temozolomide or everolimus. Based on the immunohistochemistry panel, an immunohistochemistry prognostic score (IPS) was developed. RESULTS: In patients treated with capecitabine and temozolomide, low IPS was an independent prognostic factor for progression-free-survival and overall-survival. Similar findings were observed with highest IPS for overall-survival in patients treated with everolimus. CONCLUSION: From our knowledge, it is the first time that a simple IPS could be useful to predict outcome for patients with metastatic pancreatic neuroendocrine tumors treated with everolimus or capecitabine and temozolomide.

An Interdisciplinary Approach to Optimize the Care of Transitioning Adolescents and Young Adults with CKD.

Adolescents and young adults (AYAs) with CKD or end-stage kidney disease (ESKD) have unique medical, dental, psychosocial, neurocognitive, and academic needs and require close interdisciplinary collaboration to optimize their care. The etiology of CKD in AYAs is diverse compared to older adults. With their continuously improved survival, AYAs must start preparation for health-care transition (HCT) from pediatric- to adult-focused health care in the pediatric setting and it must continue at the adult-focused setting, given that their brain maturation and self-management skill acquisition occur until their mid-20s. While the growth and physical maturation of most visible body parts occur before 18 years of age, the prefrontal cortex of the brain, where reasoning, impulse control, and other higher executive functions reside, matures around 25 years of age. The HCT process must be monitored using patient- and caregiver-measuring tools to guide interventions. The HCT process becomes more complex when patients and/or caregivers have a language barrier, different cultural beliefs, or lower literacy levels. In this article, we discuss the unique comorbidities of pediatric-onset CKD/ESKD, provide information for a planned HCT preparation, and suggest interdisciplinary coordination as well as cultural and literacy-appropriate activities to achieve optimal patient outcomes.

The effect of caffeine on cognitive performance is influenced by CYP1A2 but not ADORA2A genotype, yet neither genotype affects exercise performance in healthy adults.

PURPOSE: To determine the influence of two commonly occurring genetic polymorphisms on exercise, cognitive performance, and caffeine metabolism, after caffeine ingestion. METHODS: Eighteen adults received caffeine or placebo (3 mg kg-1) in a randomised crossover study, with measures of endurance exercise (15-min cycling time trial; 70-min post-supplementation) and cognitive performance (psychomotor vigilance test; PVT; pre, 50 and 95-min post-supplementation). Serum caffeine and paraxanthine were measured (pre, 30 and 120-min post-supplementation), and polymorphisms in ADORA2A (rs5751876) and CYP1A2 (rs762551) genes analysed. RESULTS: Caffeine enhanced exercise performance (P < 0.001), but effects were not different between participants with ADORA2A 'high' (n = 11) vs. 'low' (n = 7) sensitivity genotype (+ 6.4 ± 5.8 vs. + 8.2 ± 6.8%), or CYP1A2 'fast' (n = 10) vs. 'slow' (n = 8) metabolism genotype (+ 7.2 ± 5.9 vs. + 7.0 ± 6.7%, P > 0.05). Caffeine enhanced PVT performance (P < 0.01). The effect of caffeine was greater for CYP1A2 'fast' vs. 'slow' metabolisers for reaction time during exercise (- 18 ± 9 vs. - 1.0 ± 11 ms); fastest 10% reaction time at rest (- 18 ± 11 vs. - 3 ± 15 ms) and lapses at rest (- 3.8 ± 2.7 vs. + 0.4 ± 0.9) (P < 0.05). There were no PVT differences between ADORA2A genotypes (P > 0.05). Serum caffeine and paraxanthine responses were not different between genotypes (P > 0.05). CONCLUSION: Caffeine enhanced CYP1A2 'fast' metabolisers' cognitive performance more than 'slow' metabolisers. No other between-genotype differences emerged for the effect of caffeine on exercise or cognitive performance, or metabolism.

Maternal and Neonatal Hair Cortisol Levels and Psychological Stress Are Associated With Onset of Secretory Activation of Human Milk Production.

BACKGROUND: Several factors can influence the production of mothers' own milk. PURPOSE: To assess the influence of maternal psychological stress, maternal cortisol levels, and neonatal hair cortisol levels on timing of secretory activation. METHODS: A prospective study was conducted at 2 public health centers in Andalusia, Spain. Participants were 60 pregnant women and their 60 neonates. Hair cortisol levels and psychological stress (pregnancy-specific stress [Prenatal Distress Questionnaire, PDQ] and perceived stress [Perceived Stress Scale, PSS]) were evaluated during the third trimester and the postpartum period. This study was part of the GESTASTRESS cohort study on the effects of stress during pregnancy. RESULTS: Higher PDQ and PSS scores (P < .05) in the third trimester were associated with later onset of secretory activation. Higher postpartum maternal hair cortisol levels were associated with a delayed secretory activation of mother's own milk (P < .05). IMPLICATIONS FOR RESEARCH: Future studies should look at the influence of psychological stress and cortisol levels on hormones involved in mother's own milk production. IMPLICATIONS FOR PRACTICE: Neonatal nurses and other healthcare providers should be familiar with levels of neonates' exposure to maternal prenatal stress prior to birth.

Maternal hair cortisol levels affect neonatal development among women conceiving with assisted reproductive technology.

Objective: To compare stress levels throughout pregnancy in women who had conceived using assisted reproductive technology (ART) and women who had conceived naturally and somatometric values for infants. Background: Pregnant women who have received ART are exposed to high levels of stress. Methods: Ninety-one women attending a prenatal appointment at a Health Centre (Granada, Spain), and their 91 newborns participated in this study: 69 women conceiving naturally and 22 conceiving using ART. Assessment consisted of measuring hair cortisol levels, the Prenatal Distress Questionnaire, the Perceived Stress Scale and the Symptom Checklist-90-Revised. Results: Women conceiving using ART had higher cortisol levels in the first trimester than women who conceived naturally. In the third trimester, women who used ART reported higher levels of perceived stress than those who had conceived naturally. Maternal cortisol levels in the first trimester explained 32% of the variance in neonatal head circumference in the group of women who had conceived using ART. Conclusions: Women who had conceived using ART showed higher levels of cortisol in the first trimester and higher levels of perceived stress in the third trimester than women who had conceived naturally, rendering them more vulnerable to adverse outcomes. Maternal cortisol predicted the infants' development.

Investigation of the binding modes of a positively charged pillar[5]arene: internal and external guest complexation.

The selective binding behavior of a trimethylammonium-derived pillar[5]arene towards different guests in aqueous media and under neutral conditions is reported. Although it is known that this macrocycle has the capability to form complexes with guests, we anticipate that the intrinsic pillar shape of the macrocycle with two positively charged rims should allow a diversity of binding modes. The three guests were selected based on their charge and size. The inclusion binding modes and the affinity of the macrocycle to form host-guest complexes were determined by ITC and NMR techniques. We reveal the ability of a cationic water soluble pillar[5]arene to effectively complex two guest molecules, one in each rim, evidencing the diversity of binding modes. Two different structures for 1 : 1 and three for 1 : 2 complexes are reported showing the pillararene ability for internal/external binding.

Lagrangian tracking of long-lasting plastic tags: From lobster fisheries in the USA and Canada to Macaronesia.

Plastic waste from the fishing industry, particularly lobster trap identification tags from the USA and Canada, poses a significant threat to marine ecosystems due to its resilience. This study unveils a novel link between North American fisheries and the appearance of these plastic tags in Macaronesia. Collected in the Azores and Canary Islands, these tags offer a unique insight into the sources and spatio-temporal scales of marine plastic pollution. Ocean model data indicates the Labrador Current and Gulf Stream as key forces transporting these tags. Virtual particle simulations show a small fraction reaching Macaronesia (4.12 % in the Azores, 0.76 % in the Canary Islands), suggesting real ocean drift. The Azores, with more collected tags, are more susceptible, and tags can reach Macaronesia in under a year. These findings underscore the urgency of better waste management and emphasize the role of citizen science in monitoring and combating marine pollution.

Specific Multiomic Profiling in Aortic Stenosis in Bicuspid Aortic Valve Disease.

INTRODUCTION AND PURPOSE: Bicuspid aortic valve (BAV) disease is associated with faster aortic valve degeneration and a high incidence of aortic stenosis (AS). In this study, we aimed to identify differences in the pathophysiology of AS between BAV and tricuspid aortic valve (TAV) patients in a multiomics study integrating metabolomics and transcriptomics as well as clinical data. METHODS: Eighteen patients underwent aortic valve replacement due to severe aortic stenosis: 8 of them had a TAV, while 10 of them had a BAV. RNA sequencing (RNA-seq) and proton nuclear magnetic resonance spectroscopy (1H-NMR) were performed on these tissue samples to obtain the RNA profile and lipid and low-molecular-weight metabolites. These results combined with clinical data were posteriorly compared, and a multiomic profile specific to AS in BAV disease was obtained. RESULTS: H-NMR results showed that BAV patients with AS had different metabolic profiles than TAV patients. RNA-seq also showed differential RNA expression between the groups. Functional analysis helped connect this RNA pattern to mitochondrial dysfunction. Integration of RNA-seq, 1H-NMR and clinical data helped create a multiomic profile that suggested that mitochondrial dysfunction and oxidative stress are key players in the pathophysiology of AS in BAV disease. CONCLUSIONS: The pathophysiology of AS in BAV disease differs from patients with a TAV and has a specific RNA and metabolic profile. This profile was associated with mitochondrial dysfunction and increased oxidative stress.

Spliceosomic dysregulation in pancreatic cancer uncovers splicing factors PRPF8 and RBMX as novel candidate actionable targets.

Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal cancer, characterized by late diagnosis and poor treatment response. Surgery is the only curative approach, only available to early-diagnosed patients. Current therapies have limited effects, cause severe toxicities, and minimally improve overall survival. Understanding of splicing machinery alterations in PDAC remains incomplete. Here, we comprehensively examined 59 splicing machinery components, uncovering dysregulation in pre-mRNA processing factor 8 (PRPF8) and RNA-binding motif protein X-linked (RBMX). Their downregulated expression was linked to poor prognosis and malignancy features, including tumor stage, invasion and metastasis, and associated with poorer survival and the mutation of key PDAC genes. Experimental modulation of these splicing factors in pancreatic cancer cell lines reverted their expression to non-tumor levels and resulted in decreased key tumor-related features. These results provide evidence that the splicing machinery is altered in PDAC, wherein PRPF8 and RBMX emerge as candidate actionable therapeutic targets.