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Somatic copy number alterations play a critical role in oncogenesis. Loss of chromosomal regions containing tumor suppressors can lead to collateral deletion of passenger genes. This can be exploited therapeutically if synthetic lethal partners of such passenger genes are known and represent druggable targets. Here, we report that VPS4B gene, encoding an ATPase involved in ESCRT‐dependent membrane remodeling, is such a passenger gene frequently deleted in many cancer types, notably in colorectal cancer (CRC). We observed downregulation of VPS4B mRNA and protein levels from CRC patient samples. We identified VPS4A paralog as a synthetic lethal interactor for VPS4B in vitro and in mouse xenografts. Depleting both proteins profoundly altered the cellular transcriptome and induced cell death accompanied by the release of immunomodulatory molecules that mediate inflammatory and anti‐tumor responses. Our results identify a pair of novel druggable targets for personalized oncology and provide a rationale to develop VPS4 inhibitors for precision therapy of VPS4B‐deficient cancers. Synopsis VPS4B and VPS4A paralogs are involved in the remodeling of biological membranes, a critical step for many intracellular processes. This study highlights the possibility of using synthetic lethality between these paralogs for treatment of VPS4B‐deficient cancers. VPS4B protein abundance was decreased in colorectal cancer (CRC) patient samples. A synthetic lethal phenotype was generated by simultaneous depletion of VPS4A and B in various CRC cell lines grown in vitro and in vivo . Synthetic lethality between VPS4A and B was independent of other oncogenic mutations, and conserved between human and mouse, thus of high penetrance. Simultaneous depletion of VPS4A and B caused pleiotropic effects e.g. inhibited endocytosis and cell cycle progression, and induced a stress‐associated sterile inflammatory response. DAMPs and other immunomodulatory molecules released by VPS4A+B‐depleted dying cells may favor the induction of anti‐tumor innate and adaptive immune responses. Graphical Abstract VPS4B and VPS4A paralogs are involved in the remodeling of biological membranes, a critical step for many intracellular processes. This study highlights the possibility of using synthetic lethality between these paralogs for treatment of VPS4B‐deficient cancers.

Many adaptor proteins involved in endocytic cargo transport exhibit additional functions in other cellular processes which may be either related to or independent from their trafficking roles. The endosomal adaptor protein Tollip is an example of such a multitasking regulator, as it participates in trafficking and endosomal sorting of receptors, but also in interleukin/Toll/NF-κB signaling, bacterial entry, autophagic clearance of protein aggregates and regulation of sumoylation. Here we describe another role of Tollip in intracellular signaling. By performing a targeted RNAi screen of soluble endocytic proteins for their additional functions in canonical Wnt signaling, we identified Tollip as a potential negative regulator of this pathway in human cells. Depletion of Tollip potentiates the activity of β-catenin/TCF-dependent transcriptional reporter, while its overproduction inhibits the reporter activity and expression of Wnt target genes. These effects are independent of dynamin-mediated endocytosis, but require the ubiquitin-binding CUE domain of Tollip. In Wnt-stimulated cells, Tollip counteracts the activation of β-catenin and its nuclear accumulation, without affecting its total levels. Additionally, under conditions of ligand-independent signaling, Tollip inhibits the pathway after the stage of β-catenin stabilization, as observed in human cancer cell lines, characterized by constitutive β-catenin activity. Finally, the regulation of Wnt signaling by Tollip occurs also during early embryonic development of zebrafish. In summary, our data identify a novel function of Tollip in regulating the canonical Wnt pathway which is evolutionarily conserved between fish and humans. Tollip-mediated inhibition of Wnt signaling may contribute not only to embryonic development, but also to carcinogenesis. Mechanistically, Tollip can potentially coordinate multiple cellular pathways of trafficking and signaling, possibly by exploiting its ability to interact with ubiquitin and the sumoylation machinery.

Within the endolysosomal pathway in mammalian cells, ESCRT complexes facilitate degradation of proteins residing in endosomal membranes. Here, we show that mammalian ESCRT-I restricts the size of lysosomes and promotes degradation of proteins from lysosomal membranes, including MCOLN1, a Ca 2+ channel protein. The altered lysosome morphology upon ESCRT-I depletion coincided with elevated expression of genes annotated to biogenesis of lysosomes due to prolonged activation of TFEB/TFE3 transcription factors. Lack of ESCRT-I also induced transcription of cholesterol biosynthesis genes, in response to inefficient delivery of cholesterol from endolysosomal compartments. Among factors that could possibly activate TFEB/TFE3 signaling upon ESCRT-I deficiency, we excluded lysosomal cholesterol accumulation and Ca 2+ -mediated dephosphorylation of TFEB/TFE3. However, we discovered that this activation occurs due to the inhibition of Rag GTPase–dependent mTORC1 pathway that specifically reduced phosphorylation of TFEB at S122. Constitutive activation of the Rag GTPase complex in cells lacking ESCRT-I restored S122 phosphorylation and prevented TFEB/TFE3 activation. Our results indicate that ESCRT-I deficiency evokes a homeostatic response to counteract lysosomal nutrient starvation, that is, improper supply of nutrients derived from lysosomal degradation.

Introduction: Children with psychomotor retardation demonstrate lower than average level of development, that is characteristic for the majority of population in a certain period of life. These children require specialised, coordinated and interdisciplinary therapeutic efforts. The aim of the present thesis was to assess the influence of choreotherapy on the development of children with psychomotor retardation. Material and methods: The study consisted of two measurement sessions of 4-6 years old children attending for therapeutic activities to the Early Intervention Centre in Mielec. The research tool was the Bogdanowicz Behaviour Observation Scale (SOZ-D). Basic descriptive statistics were calculated in the analysis. The Mann-Whitney U test was used to assess differences in psychomotor development between the examined group (participating in the therapeutic program which included choreotherapy) and the control group. The rate of improvement was tested by means of the ANOVA Kruskal-Wallis test. Results: There were statistically significant differences between both the examined and the control group in all subscales of the SOZ-D scale. At children participating in the experimental program the greatest improvement was noticed in emotional, social and physical spheres of development. Conclusions: The applied therapeutic program had a beneficial effect on stimulating the development of emotional, social and physical spheres of children with psychomotor retardation. The present thesis demonstrates the usefulness of this program, and consequently implies the need of propagating the therapy in other educational and care centres for children.

Somatic copy number alterations play a critical role in oncogenesis. Loss of chromosomal regions containing tumor suppressors can lead to collateral deletion of passenger genes. This can be exploited therapeutically if synthetic lethal partners of such passenger genes are known and represent druggable targets. Here, we report that VPS4B gene, encoding an ATPase involved in ESCRT‐dependent membrane remodeling, is such a passenger gene frequently deleted in many cancer types, notably in colorectal cancer (CRC). We observed downregulation of VPS4B mRNA and protein levels from CRC patient samples. We identified VPS4A paralog as a synthetic lethal interactor for VPS4B in vitro and in mouse xenografts. Depleting both proteins profoundly altered the cellular transcriptome and induced cell death accompanied by the release of immunomodulatory molecules that mediate inflammatory and anti‐tumor responses. Our results identify a pair of novel druggable targets for personalized oncology and provide a rationale to develop VPS4 inhibitors for precision therapy of VPS4B‐deficient cancers.

Molecular details of how endocytosis contributes to oncogenesis remain elusive. Our in silico analysis of colorectal cancer (CRC) patients revealed stage-dependent alterations in the expression of 113 endocytosis-related genes. Among them transcription of the Endosomal Sorting Complex Required for Transport (ESCRT)-I component VPS37B was decreased in the advanced stages of CRC. Expression of other ESCRT-I core subunits remained unchanged in the investigated dataset. We analyzed an independent cohort of CRC patients showing also reduced VPS37A mRNA and protein abundance. Transcriptomic profiling of CRC cells revealed non-redundant functions of Vps37 proteins. Knockdown of VPS37A and VPS37B triggered p21-mediated inhibition of cell proliferation and sterile inflammatory response driven by the Nuclear Factor (NF)-kappaB transcription factor and associated with mitogen-activated protein kinase signaling. Co-silencing of VPS37C further potentiated activation of these independently induced processes. The type and magnitude of transcriptional alterations correlated with the differential ESCRT-I stability upon individual and concurrent Vps37 depletion. Our study provides novel insights into cancer cell biology by describing cellular stress responses that are associated with ESCRT-I destabilization, which might occur in CRC patients.

Within the endolysosomal pathway in mammalian cells, ESCRT complexes facilitate degradation of proteins residing in endosomal membranes. Recent studies revealed that yeast ESCRT machinery also sorts ubiquitinated proteins from the vacuolar membrane for degradation in the vacuole lumen. However, whether mammalian ESCRTs perform a similar function at lysosomes remained unknown. Here, we show that ESCRT-I restricts the size of lysosomes and promotes degradation of proteins from lysosomal membranes, including MCOLN1, a Ca2+ channel protein. Upon ESCRT-I depletion, the lysosomal accumulation of non-degraded proteins coincided with elevated expression of genes annotated to cholesterol biosynthesis and biogenesis of lysosomes, indicative of response to lysosomal stress. Accordingly, the lack of ESCRT-I promoted abnormal cholesterol accumulation in lysosomes and activated TFEB/TFE3 transcription factors. Finally, we discovered that in contrast to basal TFEB/TFE3 signaling that depended on the availability of exogenous lipids, the stress-induced activation of this pathway was Ca2+-MCOLN1-dependent. Hence, we provide evidence that ESCRT-I is crucial for maintaining lysosomal homeostasis and we elucidate mechanisms distinguishing basal from lysosomal stress-induced TFEB/TFE3 signaling.

Shoot cultures of Nepeta curviflora were used to test the effect of 6-benzylaminopurine (BAP) and riboside 6-benzylaminopurine (r-BAP) on the growth and production of phenolic compounds. The best multiplication was achieved on agar Murashige and Skoog medium (MS) with r-BAP (1.5 mg/L), where 74% of explants produced about eight axillary shoots. The fresh weight (FW) was about 0.6 g/tube (24 g/L) and the dry weight (DW) was about 0.04 g/tube (2 g/L). To scale up the culture, the shoot culture was grown for the first time in a nutrient sprinkle bioreactor. After 4 weeks of culture, the multiplication rate (8.13) was higher than that observed in glass tubes (8.03). The fresh biomass was 113.2 g/bioreactor (75.5 g/L) and the dry mass was 14 g/bioreactor (9.3 g/L). Extracts from obtained plant material were analyzed by the UPLC/DAD/qTOF-MS technique. A total of 32 phenolic compounds were identified and quantified. The total content of compounds ranged from 600 to 1400 mg/100 g of dry weight (DW), depending on the culture conditions. In the profile of shoot cultures, rosmarinic acid was dominant, whereas prolithospermic acid was mostly noted in extract from aerial parts of the plant obtained from the field.

Urban forest ecosystems face increasing abiotic stress due to climate change and anthropogenic pressure, requiring robust indicators of plant adaptation. Hedera helix L., a widespread liana with shoot dimorphism, plays a key role in temperate urban woodlands but remains understudied in terms of multilevel responses to environmental gradients. We assessed vegetative and generative shoots with healthy or damaged leaves across heterogeneous urban forest sites to identify functional responses to volumetric water content (VWC), temperature (t), electrical conductivity (ES), and daily light integral (DLI). VWC and DLI emerged as the key drivers of trait variability. The multilevel adaptive trait responses demonstrated the ecological flexibility of a dominant urban liana. We developed a novel Integrative Ecological Index (IEI) based on normalized trait sub-indices, which captures multilevel plant responses to environmental stress and enables quantitative assessment of urban habitat conditions. IEI was highest in healthy vegetative shoots and lowest in damaged generative shoots. Our findings demonstrate the ecological plasticity of H. helix and its potential as a sensitive bioindicator. This interdisciplinary study provides a scalable trait-based tool to evaluate adaptive capacity and microhabitat conditions in heterogeneous urban environments.