High-flow nasal cannula in Spanish Pediatric Intensive Care Services: A national web survey about its use and indications.

OBJECTIVE: To describe the high-flow nasal cannula (HFNC) indications in the Spanish pediatric critical care units (PICUs). DESIGN: Descriptive cross-sectional observational study. SETTING: Electronic survey among members of the Spanish Society of Pediatric Intensive Care (SECIP). It was sent weekly from April 10, 2023, to May 21, 2023. PARTICIPANTS: All SECIP members. INTERVENTIONS: None. MAIN VARIABLES OF INTEREST: The questions focused on workplace, years of experience, use or non-use of HFNC, justification and expectations regarding its application, starting point within each center, clinical criteria for indication, existence of clinical guidelines, evaluation during its use, and criteria and mode of withdrawal. RESULTS: Two hundred and two participants, 176 were from Spain. Of these, 87/176 had over ten years of experience. One hundred sixty two use HFNC and 66/162 have HFNC clinical guidelines. Acute bronchiolitis (138/162) and respiratory assistance after extubation (106/56) are the two main indications. For 62/162 HFNC may reduce therapeutic escalation. Neuromuscular diseases (105/162) and anatomical airway diseases (135/162) are the two main contraindications. The reasons to do not use HFNC were the absence of evidence about it effectiveness (8/14) and its inadequate cost/effectiveness balance (8/14). CONCLUSIONS: A majority of Spanish pediatric intensivists use HFNC. Its application and withdrawal appears to be primarily based on clinical experience. Besides, those who use HFNC are aware of its limitations and the lack of evidence in some cases. It is necessary to develop single-center and multicenter studies to elucidate the effectiveness of this therapy in the context of critically ill children.

Comprehensive study of mechanical power in controlled mechanical ventilation: Prevalence of elevated mechanical power and component analysis / Estudio Integral de la Potencia Mecánica en la Ventilación Mecánica Controlada: Prevalencia de Potencia Mecánica Elevada y Análisis de Componentes

Objective To determine the prevalence of elevated mechanical power (MP) values (>17J/min) used in routine clinical practice. Design Observational, descriptive, cross-sectional, analytical, multicenter, international study conducted on November 21, 2019, from 8:00 AM to 3:00 PM. NCT03936231. Setting One hundred thirty-three Critical Care Units. Patients Patients receiving invasive mechanical ventilation for any cause. Interventions None. Main variables of interest Mechanical power. Results A population of 372 patients was analyzed. PM was significantly higher in patients under pressure-controlled ventilation (PC) compared to volume-controlled ventilation (VC) (19.20±8.44J/min vs. 16.01±6.88J/min; p<0.001), but the percentage of patients with PM>17J/min was not different (41% vs. 35%, respectively; p=0.382). The best models according to AICcw expressing PM for patients in VC are described as follows: Surrogate Strain (Driving Pressure) + PEEP+Surrogate Strain Rate (PEEP/Flow Ratio) + Respiratory Rate. For patients in PC, it is defined as: Surrogate Strain (Expiratory Tidal Volume/PEEP) + PEEP+Surrogate Strain Rate (Surrogate Strain/Ti) + Respiratory Rate+Expiratory Tidal Volume+Ti. Conclusions A substantial proportion of mechanically ventilated patients may be at risk of experiencing elevated levels of mechanical power. Despite observed differences in mechanical power values between VC and PC ventilation, they did not result in a significant disparity in the prevalence of high mechanical power values. (AU)

Objetivo Determinar la prevalencia de valores elevados de potencia mecánica (PM) (>17J/min) utilizados en la práctica clínica habitual. Diseño estudio observacional, descriptivo de corte transversal, analítico, multicéntrico e internacional, realizado el 21 de noviembre de 2019 en horario de 8 a 15 horas. NCT03936231. Ámbito Ciento treinta y tres Unidad de Cuidados Críticos. Pacientes pacientes que recibirán ventilación mecánica por cualquier causa. Intervenciones ninguna Variables de interés principales Potencia mecánica. Resultados se analizaron 372 enfermos. La PM fue significativamente mayor en pacientes en ventilación controlada por presión (PC) que en ventilación controlada por volumen (VC) (19,20+8,44J/min frente a 16,01+6,88J/min; p<0,001), pero el porcentaje de pacientes con PM>17J/min no fue diferente (41% frente a 35% respectivamente; p=0,382). Los mejores modelos según AICcw que expresan la PM para los enfermos en VC se decribe como: Strain subrogante (Presión de conducción) + PEEP+Strain Rate subrogante (PEEP/cociente de flujo) + Frecuencia respiratoria. Para los enfermos en PC se define como: Strain subrogante (Volumen tidal expiratorio/PEEP) + PEEP+Strain Rate subrogante (Strain subrogante/Ti) + Frecuencia respiratoria+Expiratory Tidal Volumen+Ti. Conclusiones Gran parte de los pacientes en ventilación mecánica en condiciones de práctica clínica habitual reciben niveles de potencia mecánica peligrosos. A pesar de las diferencias observadas en los valores de potencia mecánica entre la ventilación VC y PC, este porcentaje de riesgo fue similar en PC y VC. (AU)

Paradoxical activation of oncogenic signaling as a cancer treatment strategy.

Cancer homeostasis depends on a balance between activated oncogenic pathways driving tumorigenesis and engagement of stress-response programs that counteract the inherent toxicity of such aberrant signaling. While inhibition of oncogenic signaling pathways has been explored extensively, there is increasing evidence that overactivation of the same pathways can also disrupt cancer homeostasis and cause lethality. We show here that inhibition of Protein Phosphatase 2A (PP2A) hyperactivates multiple oncogenic pathways and engages stress responses in colon cancer cells. Genetic and compound screens identify combined inhibition of PP2A and WEE1 as synergistic in multiple cancer models by collapsing DNA replication and triggering premature mitosis followed by cell death. This combination also suppressed the growth of patient-derived tumors in vivo. Remarkably, acquired resistance to this drug combination suppressed the ability of colon cancer cells to form tumors in vivo. Our data suggest that paradoxical activation of oncogenic signaling can result in tumor suppressive resistance.

1,4-dihydroxy quininib activates ferroptosis pathways in metastatic uveal melanoma and reveals a novel prognostic biomarker signature.

Uveal melanoma (UM) is an ocular cancer, with propensity for lethal liver metastases. When metastatic UM (MUM) occurs, as few as 8% of patients survive beyond two years. Efficacious treatments for MUM are urgently needed. 1,4-dihydroxy quininib, a cysteinyl leukotriene receptor 1 (CysLT1) antagonist, alters UM cancer hallmarks in vitro, ex vivo and in vivo. Here, we investigated the 1,4-dihydroxy quininib mechanism of action and its translational potential in MUM. Proteomic profiling of OMM2.5 cells identified proteins differentially expressed after 1,4-dihydroxy quininib treatment. Glutathione peroxidase 4 (GPX4), glutamate-cysteine ligase modifier subunit (GCLM), heme oxygenase 1 (HO-1) and 4 hydroxynonenal (4-HNE) expression were assessed by immunoblots. Biliverdin, glutathione and lipid hydroperoxide were measured biochemically. Association between the expression of a specific ferroptosis signature and UM patient survival was performed using public databases. Our data revealed that 1,4-dihydroxy quininib modulates the expression of ferroptosis markers in OMM2.5 cells. Biochemical assays validated that GPX4, biliverdin, GCLM, glutathione and lipid hydroperoxide were significantly altered. HO-1 and 4-HNE levels were significantly increased in MUM tumor explants from orthotopic patient-derived xenografts (OPDX). Expression of genes inhibiting ferroptosis is significantly increased in UM patients with chromosome 3 monosomy. We identified IFerr, a novel ferroptosis signature correlating with UM patient survival. Altogether, we demontrated that in MUM cells and tissues, 1,4-dihydroxy quininib modulates key markers that induce ferroptosis, a relatively new type of cell death driven by iron-dependent peroxidation of phospholipids. Furthermore, we showed that high expression of specific genes inhibiting ferroptosis is associated with a worse UM prognosis, thus, the IFerr signature is a potential prognosticator for which patients develop MUM. All in all, ferroptosis has potential as a clinical biomarker and therapeutic target for MUM.

Reduced Match Exposure in the Previous 2 Matches Accounts for Hamstring Muscle Injury Incidence in Professional Football Players.

BACKGROUND: Hamstring strain injuries are one of the most prevalent injuries in football (soccer). We examined the influence of accumulated match-play exposure on the occurrence of hamstring strain injury in professional football from 2 teams (Spanish 1st Division, LaLiga) over 3 seasons, and determined specific cut-off points as indicators of injury risk. HYPOTHESIS: Overloaded players would be more likely to sustain a hamstring injury. STUDY DESIGN: Prospective, controlled, observational study. LEVEL OF EVIDENCE: Level 2b. METHODS: Playing time, total running distance, and high-speed running (>24 km/h) distance during official matches of players that sustained a hamstring injury were compared with uninjured, paired controls. Cumulative playing time and running performance of 4 matches before the injury was computed. Relative risk (RR) of injury occurrence was estimated by generalized estimating equations. Diagnostic accuracy was determined by receiver operating characteristics and the area under the curve. RESULTS: Thirty-seven hamstring strain injuries occurred, representing 23 ± 18 absence days per injury. Thirty-seven controls (uninjured players) were used as comparators. Low match-play exposures during 1 and 2 matches before injury were likely to explain injury occurrence (RR: 14-53%; P < 0.01). Metrics from the match before the hamstring muscle strain demonstrated the best accuracy to predict injury occurrence: high-speed running distance ≤328 m (sensitivity, 64%; specificity, 84%), playing time ≤64 min (sensitivity, 36%; specificity, 97%), and running distance ≤5.8 km (sensitivity, 39%; specificity, 97%). CONCLUSION: Relatively reduced competitive exposure in the previous 2 matches was associated with higher hamstring injury risk in professional football players. CLINICAL RELEVANCE: Screening simple metrics such as the accumulated match exposure during official matches and considering specific cut-off points for some running variables may be good indicators of injury risk and may assist in better individual injury management in professional soccer players.

Comprehensive study of mechanical power in controlled mechanical ventilation: Prevalence of elevated mechanical power and component analysis.

OBJECTIVE: To determine the prevalence of elevated mechanical power (MP) values (>17J/min) used in routine clinical practice. DESIGN: Observational, descriptive, cross-sectional, analytical, multicenter, international study conducted on November 21, 2019, from 8:00 AM to 3:00 PM. NCT03936231. SETTING: One hundred thirty-three Critical Care Units. PATIENTS: Patients receiving invasive mechanical ventilation for any cause. INTERVENTIONS: None. MAIN VARIABLES OF INTEREST: Mechanical power. RESULTS: A population of 372 patients was analyzed. PM was significantly higher in patients under pressure-controlled ventilation (PC) compared to volume-controlled ventilation (VC) (19.20±8.44J/min vs. 16.01±6.88J/min; p<0.001), but the percentage of patients with PM>17J/min was not different (41% vs. 35%, respectively; p=0.382). The best models according to AICcw expressing PM for patients in VC are described as follows: Surrogate Strain (Driving Pressure) + PEEP+Surrogate Strain Rate (PEEP/Flow Ratio) + Respiratory Rate. For patients in PC, it is defined as: Surrogate Strain (Expiratory Tidal Volume/PEEP) + PEEP+Surrogate Strain Rate (Surrogate Strain/Ti) + Respiratory Rate+Expiratory Tidal Volume+Ti. CONCLUSIONS: A substantial proportion of mechanically ventilated patients may be at risk of experiencing elevated levels of mechanical power. Despite observed differences in mechanical power values between VC and PC ventilation, they did not result in a significant disparity in the prevalence of high mechanical power values.

Expanding a precision medicine platform for malignant peripheral nerve sheath tumors: New patient-derived orthotopic xenografts, cell lines and tumor entities.

Malignant peripheral nerve sheath tumors (MPNSTs) are aggressive soft-tissue sarcomas with a poor survival rate, presenting either sporadically or in the context of neurofibromatosis type 1 (NF1). The histological diagnosis of MPNSTs can be challenging, with different tumors exhibiting great histological and marker expression overlap. This heterogeneity could be partly responsible for the observed disparity in treatment response due to the inherent diversity of the preclinical models used. For several years, our group has been generating a large patient-derived orthotopic xenograft (PDOX) MPNST platform for identifying new precision medicine treatments. Herein, we describe the expansion of this platform using six primary tumors clinically diagnosed as MPNSTs, from which we obtained six additional PDOX mouse models and three cell lines, thus generating three pairs of in vitro-in vivo models. We extensively characterized these tumors and derived preclinical models, including genomic, epigenomic, and histological analyses. Tumors were reclassified after these analyses: three remained as MPNSTs (two being classic MPNSTs), one was a melanoma, another was a neurotrophic tyrosine receptor kinase (NTRK)-rearranged spindle cell neoplasm, and, finally, the last was an unclassifiable tumor bearing neurofibromin-2 (NF2) inactivation, a neuroblastoma RAS viral oncogene homolog (NRAS) oncogenic mutation, and a SWI/SNF-related matrix-associated actin-dependent regulator of chromatin (SMARCA4) heterozygous truncated variant. New cell lines and PDOXs faithfully recapitulated histology, marker expression, and genomic characteristics of the primary tumors. The diversity in tumor identity and their specific associated genomic alterations impacted treatment responses obtained when we used the new cell lines for testing compounds against known altered pathways in MPNSTs. In summary, we present here an extension of our MPNST precision medicine platform, with new PDOXs and cell lines, including tumor entities confounded as MPNSTs in a real clinical scenario. This platform may constitute a useful tool for obtaining correct preclinical information to guide MPNST clinical trials.

IκB kinase-α coordinates BRD4 and JAK/STAT signaling to subvert DNA damage-based anticancer therapy.

Activation of the IκB kinase (IKK) complex has recurrently been linked to colorectal cancer (CRC) initiation and progression. However, identification of downstream effectors other than NF-κB has remained elusive. Here, analysis of IKK-dependent substrates in CRC cells after UV treatment revealed that phosphorylation of BRD4 by IKK-α is required for its chromatin-binding at target genes upon DNA damage. Moreover, IKK-α induces the NF-κB-dependent transcription of the cytokine LIF, leading to STAT3 activation, association with BRD4 and recruitment to specific target genes. IKK-α abrogation results in defective BRD4 and STAT3 functions and consequently irreparable DNA damage and apoptotic cell death upon different stimuli. Simultaneous inhibition of BRAF-dependent IKK-α activity, BRD4, and the JAK/STAT pathway enhanced the therapeutic potential of 5-fluorouracil combined with irinotecan in CRC cells and is curative in a chemotherapy-resistant xenograft model. Finally, coordinated expression of LIF and IKK-α is a poor prognosis marker for CRC patients. Our data uncover a functional link between IKK-α, BRD4, and JAK/STAT signaling with clinical relevance.

A biobank of pediatric patient-derived-xenograft models in cancer precision medicine trial MAPPYACTS for relapsed and refractory tumors.

Pediatric patients with recurrent and refractory cancers are in most need for new treatments. This study developed patient-derived-xenograft (PDX) models within the European MAPPYACTS cancer precision medicine trial (NCT02613962). To date, 131 PDX models were established following heterotopical and/or orthotopical implantation in immunocompromised mice: 76 sarcomas, 25 other solid tumors, 12 central nervous system tumors, 15 acute leukemias, and 3 lymphomas. PDX establishment rate was 43%. Histology, whole exome and RNA sequencing revealed a high concordance with the primary patient's tumor profile, human leukocyte-antigen characteristics and specific metabolic pathway signatures. A detailed patient molecular characterization, including specific mutations prioritized in the clinical molecular tumor boards are provided. Ninety models were shared with the IMI2 ITCC Pediatric Preclinical Proof-of-concept Platform (IMI2 ITCC-P4) for further exploitation. This PDX biobank of unique recurrent childhood cancers provides an essential support for basic and translational research and treatments development in advanced pediatric malignancies.

Epigenetic inactivation of the 5-methylcytosine RNA methyltransferase NSUN7 is associated with clinical outcome and therapeutic vulnerability in liver cancer.

BACKGROUND: RNA modifications are important regulators of transcript activity and an increasingly emerging body of data suggests that the epitranscriptome and its associated enzymes are altered in human tumors. METHODS: Combining data mining and conventional experimental procedures, NSUN7 methylation and expression status was assessed in liver cancer cell lines and primary tumors. Loss-of-function and transfection-mediated recovery experiments coupled with RNA bisulfite sequencing and proteomics determined the activity of NSUN7 in downstream targets and drug sensitivity. RESULTS: In this study, the initial screening for genetic and epigenetic defects of 5-methylcytosine RNA methyltransferases in transformed cell lines, identified that the NOL1/NOP2/Sun domain family member 7 (NSUN7) undergoes promoter CpG island hypermethylation-associated with transcriptional silencing in a cancer-specific manner. NSUN7 epigenetic inactivation was common in liver malignant cells and we coupled bisulfite conversion of cellular RNA with next-generation sequencing (bsRNA-seq) to find the RNA targets of this poorly characterized putative RNA methyltransferase. Using knock-out and restoration-of-function models, we observed that the mRNA of the coiled-coil domain containing 9B (CCDC9B) gene required NSUN7-mediated methylation for transcript stability. Most importantly, proteomic analyses determined that CCDC9B loss impaired protein levels of its partner, the MYC-regulator Influenza Virus NS1A Binding Protein (IVNS1ABP), creating sensitivity to bromodomain inhibitors in liver cancer cells exhibiting NSUN7 epigenetic silencing. The DNA methylation-associated loss of NSUN7 was also observed in primary liver tumors where it was associated with poor overall survival. Interestingly, NSUN7 unmethylated status was enriched in the immune active subclass of liver tumors. CONCLUSION: The 5-methylcytosine RNA methyltransferase NSUN7 undergoes epigenetic inactivation in liver cancer that prevents correct mRNA methylation. Furthermore, NSUN7 DNA methylation-associated silencing is associated with clinical outcome and distinct therapeutic vulnerability.

CDK6 is activated by the atypical cyclin I to promote E2F-mediated gene expression and cancer cell proliferation.

Cyclin-dependent kinases (CDKs), together with their cyclin partners, are the master cell cycle regulators. Remarkably, the cyclin family was extended to include atypical cyclins, characterized by distinctive structural features, but their partner CDKs remain elusive. Here, we conducted a yeast two-hybrid screen to identify new atypical cyclin-CDK complexes. We identified 10 new complexes, including a complex between CDK6 and cyclin I (CCNI), which was found to be active against retinoblastoma protein. CCNI upregulation increased the proliferation of breast cancer cells in vitro and in vivo, with a magnitude similar to that seen upon cyclin D upregulation, an effect that was abrogated by CDK6 silencing or palbociclib treatment. In line with these findings, CCNI downregulation led to a decrease in cell number and a reduction in the percentage of cells reaching S phase. Finally, CCNI upregulation correlated with the high expression of E2F target genes in large panels of cancer cell lines and tissue samples from breast cancer patients. In conclusion, we unveil CCNI as a new player in the pathways that activate CDK6, enriching the wiring of cell cycle control.

Generation of human iPSC-derived neurofibromaspheres for in vitro and in vivo uses.

Neurofibromas are benign peripheral nervous system tumors associated with neurofibromatosis type 1, which originate from NF1(-/-) Schwann cell precursors. We describe a protocol to generate neurofibromaspheres by differentiating NF1(-/-) Schwann cells from induced pluripotent stem cells and combining them with neurofibroma primary fibroblasts. We also describe the development of neurofibroma-like tumors when neurofibromaspheres are engrafted in the sciatic nerve of nude mice. This model constitutes a versatile platform for drug screening and the study of neurofibroma biology. For complete details on the use and execution of this protocol, please refer to Mazuelas et al. (2022).1.

MmCMS: mouse models' consensus molecular subtypes of colorectal cancer.

BACKGROUND: Colorectal cancer (CRC) primary tumours are molecularly classified into four consensus molecular subtypes (CMS1-4). Genetically engineered mouse models aim to faithfully mimic the complexity of human cancers and, when appropriately aligned, represent ideal pre-clinical systems to test new drug treatments. Despite its importance, dual-species classification has been limited by the lack of a reliable approach. Here we utilise, develop and test a set of options for human-to-mouse CMS classifications of CRC tissue. METHODS: Using transcriptional data from established collections of CRC tumours, including human (TCGA cohort; n = 577) and mouse (n = 57 across n = 8 genotypes) tumours with combinations of random forest and nearest template prediction algorithms, alongside gene ontology collections, we comprehensively assess the performance of a suite of new dual-species classifiers. RESULTS: We developed three approaches: MmCMS-A; a gene-level classifier, MmCMS-B; an ontology-level approach and MmCMS-C; a combined pathway system encompassing multiple biological and histological signalling cascades. Although all options could identify tumours associated with stromal-rich CMS4-like biology, MmCMS-A was unable to accurately classify the biology underpinning epithelial-like subtypes (CMS2/3) in mouse tumours. CONCLUSIONS: When applying human-based transcriptional classifiers to mouse tumour data, a pathway-level classifier, rather than an individual gene-level system, is optimal. Our R package enables researchers to select suitable mouse models of human CRC subtype for their experimental testing.

Oxidative stress-induced FAK activation contributes to uterine serous carcinoma aggressiveness.

Uterine serous carcinoma (USC) is an aggressive form of endometrial cancer (EC), characterized by its high propensity for metastases. In fact, while endometrioid endometrial carcinoma (EEC), which accounts for 85% of EC, presents a good prognosis, USC is the most frequently fatal. Herein, we used for the first time a peptide-based tyrosine-kinase-activity profiling approach to quantify the changes in tyrosine kinase activation between USC and EEC. Among the tyrosine kinases highly activated in USC, we identified focal adhesion kinase (FAK). We conducted mechanistic studies using cellular models. In a USC cell line, targeting FAK either by inhibitors PF-573228 and defactinib (VS-6063) or by gene silencing limits 3D cell growth and reduces cell migration. Moreover, results from our studies suggest that oxidative stress is increased in USC tumors compared to EEC ones. Reactive oxygen species (ROS) induce tyrosine phosphorylation of FAK and a concomitant tyrosine phosphorylation of paxillin, a mediator of FAK signal transduction. Mechanistically, by tracking hundreds of individual cells per condition, we show that ROS increased cell distance and migration velocity, highlighting the role of ROS-FAK-PAX signaling in cell migration. Both defactinib and ROS scavenger N-acetylcysteine (NAC) revert this effect, pointing toward ROS as potential culprits for the increase in USC cell motility. A proof of concept of the role of FAK in controlling cell growth was obtained in in vivo experiments using cancer-tissue-originated spheroids (CTOS) and a patient-derived orthotopic xenograft model (orthoxenograft/PDOX). Defactinib reduces cell proliferation and protein oxidation, supporting a pro-tumoral antioxidant role of FAK, whereas antioxidant NAC reverts FAK inhibitor effects. Overall, our data points to ROS-mediated FAK activation in USC as being responsible for the poor prognosis of this tumor type and emphasize the potential of FAK inhibition for USC treatment.

Multivariate Probabilistic Range Queries for Scalable Interactive 3D Visualization.

Large-scale scientific data, such as weather and climate simulations, often comprise a large number of attributes for each data sample, like temperature, pressure, humidity, and many more. Interactive visualization and analysis require filtering according to any desired combination of attributes, in particular logical AND operations, which is challenging for large data and many attributes. Many general data structures for this problem are built for and scale with a fixed number of attributes, and scalability of joint queries with arbitrary attribute subsets remains a significant problem. We propose a flexible probabilistic framework for multivariate range queries that decouples all attribute dimensions via projection, allowing any subset of attributes to be queried with full efficiency. Moreover, our approach is output-sensitive, mainly scaling with the cardinality of the query result rather than with the input data size. This is particularly important for joint attribute queries, where the query output is usually much smaller than the whole data set. Additionally, our approach can split query evaluation between user interaction and rendering, achieving much better scalability for interactive visualization than the previous state of the art. Furthermore, even when a multi-resolution strategy is used for visualization, queries are jointly evaluated at the finest data granularity, because our framework does not limit query accuracy to a fixed spatial subdivision.

Generation and Integrated Analysis of Advanced Patient-Derived Orthoxenograft Models (PDOX) for the Rational Assessment of Targeted Therapies in Endometrial Cancer.

Clinical management of endometrial cancer (EC) is handicapped by the limited availability of second line treatments and bona fide molecular biomarkers to predict recurrence. These limitations have hampered the treatment of these patients, whose survival rates have not improved over the last four decades. The advent of coordinated studies such as The Cancer Genome Atlas Uterine Corpus Endometrial Carcinoma (TCGA_UCEC) has partially solved this issue, but the lack of proper experimental systems still represents a bottleneck that precludes translational studies from successful clinical testing in EC patients. Within this context, the first study reporting the generation of a collection of endometrioid-EC-patient-derived orthoxenograft (PDOX) mouse models is presented that is believed to overcome these experimental constraints and pave the way toward state-of-the-art precision medicine in EC. The collection of primary tumors and derived PDOXs is characterized through an integrative approach based on transcriptomics, mutational profiles, and morphological analysis; and it is demonstrated that EC tumors engrafted in the mouse uterus retain the main molecular and morphological features from analogous tumor donors. Finally, the molecular properties of these tumors are harnessed to assess the therapeutic potential of trastuzumab, a human epidermal growth factor receptor 2 (HER2) inhibitor with growing interest in EC, using patient-derived organotypic multicellular tumor spheroids and in vivo experiments.

Inter-limb asymmetry in youth elite soccer players: Effect of loading conditions.

The presence of inter-limb asymmetries can influence strength performance and represent an injury risk factor for team sport athletes. The present study aimed to investigate the effects of changes in resistance loads using different assessment modalities on the magnitude and the direction of inter-limb asymmetry within the same leg. Fifteen young elite soccer players from the same professional academy performed rear-foot-elevated-split-squat-test at different loading conditions (body mass with no overload, 25% of body mass, 50% of body mass 50%), isokinetic knee flexor (concentric 30°·s-1, concentric 60°·s-1, eccentric 90°·s-1) and extensor (concentric 60°·s-1, eccentric 60°·s-1). The outcomes from the agreement analyses suggested moderate level agreement between body mass vs body mass 25% (Kappa = 0.46), with no agreement or fair agreement for the other between-assessment comparison. Our results demonstrated that the magnitude and direction of within-limb strength imbalances were inconsistent when compared within the same assessment under different resistance load conditions.

p53 wild-type colorectal cancer cells that express a fetal gene signature are associated with metastasis and poor prognosis.

Current therapy against colorectal cancer (CRC) is based on DNA-damaging agents that remain ineffective in a proportion of patients. Whether and how non-curative DNA damage-based treatment affects tumor cell behavior and patient outcome is primarily unstudied. Using CRC patient-derived organoids (PDO)s, we show that sublethal doses of chemotherapy (CT) does not select previously resistant tumor populations but induces a quiescent state specifically to TP53 wildtype (WT) cancer cells, which is linked to the acquisition of a YAP1-dependent fetal phenotype. Cells displaying this phenotype exhibit high tumor-initiating and metastatic activity. Nuclear YAP1 and fetal traits are present in a proportion of tumors at diagnosis and predict poor prognosis in patients carrying TP53 WT CRC tumors. We provide data indicating the higher efficacy of CT together with YAP1 inhibitors for eradication of therapy resistant TP53 WT cancer cells. Together these results identify fetal conversion as a useful biomarker for patient prognosis and therapy prescription.

A High-Throughput Screening Platform Identifies Novel Combination Treatments for Malignant Peripheral Nerve Sheath Tumors.

Malignant peripheral nerve sheath tumors (MPNST) are soft-tissue sarcomas that are the leading cause of mortality in patients with Neurofibromatosis type 1 (NF1). Single chemotherapeutic agents have shown response rates ranging from 18% to 44% in clinical trials, so there is still a high medical need to identify chemotherapeutic combination treatments that improve clinical prognosis and outcome. We screened a collection of compounds from the NCATS Mechanism Interrogation PlatE (MIPE) library in three MPNST cell lines, using cell viability and apoptosis assays. We then tested whether compounds that were active as single agents were synergistic when screened as pairwise combinations. Synergistic combinations in vitro were further evaluated in patient-derived orthotopic xenograft/orthoxenograft (PDOX) athymic models engrafted with primary MPNST matching with their paired primary-derived cell line where synergism was observed. The high-throughput screening identified 21 synergistic combinations, from which four exhibited potent synergies in a broad panel of MPNST cell lines. One of the combinations, MK-1775 with Doxorubicin, significantly reduced tumor growth in a sporadic PDOX model (MPNST-SP-01; sevenfold) and in an NF1-PDOX model (MPNST-NF1-09; fourfold) and presented greater effects in TP53 mutated MPNST cell lines. The other three combinations, all involving Panobinostat (combined with NVP-BGT226, Torin 2, or Carfilzomib), did not reduce the tumor volume in vivo at noncytotoxic doses. Our results support the utility of our screening platform of in vitro and in vivo models to explore new therapeutic approaches for MPNSTs and identified that combination MK-1775 with Doxorubicin could be a good pharmacologic option for the treatment of these tumors.

Functional patient-derived organoid screenings identify MCLA-158 as a therapeutic EGFR × LGR5 bispecific antibody with efficacy in epithelial tumors.

Patient-derived organoids (PDOs) recapitulate tumor architecture, contain cancer stem cells and have predictive value supporting personalized medicine. Here we describe a large-scale functional screen of dual-targeting bispecific antibodies (bAbs) on a heterogeneous colorectal cancer PDO biobank and paired healthy colonic mucosa samples. More than 500 therapeutic bAbs generated against Wingless-related integration site (WNT) and receptor tyrosine kinase (RTK) targets were functionally evaluated by high-content imaging to capture the complexity of PDO responses. Our drug discovery strategy resulted in the generation of MCLA-158, a bAb that specifically triggers epidermal growth factor receptor degradation in leucine-rich repeat-containing G-protein-coupled receptor 5-positive (LGR5+) cancer stem cells but shows minimal toxicity toward healthy LGR5+ colon stem cells. MCLA-158 exhibits therapeutic properties such as growth inhibition of KRAS-mutant colorectal cancers, blockade of metastasis initiation and suppression of tumor outgrowth in preclinical models for several epithelial cancer types.