Biology subjects

Desterke, C.

Publications and source records attributed to Desterke, C..

15 recordsLinked to original sources

bioRxiv2026

CBX3 Deletion Overcomes Ferroptosis Resistance in Colorectal Cancer by Targeting IFNγ/SLC7A11/SLC3A2 Axis

System Xc- (SLC7A11/SLC3A2) maintains redox balance by supporting glutathione synthesis, thereby preventing lipid peroxidation and ferroptosis, and represents a promising therapeutic target in cancer. In this work, we revealed that an important epigenetic regulator, CBX3 deletion significantly increased intracellular ROS and lipid ROS level following ferroptosis inducing, particularly under IFN{gamma} stimulation, thereby sensitized CRC cells to ferroptosis in vitro using human CRC cells as well as in vivo using MC38 syngenetic tumors. RNAseq with mice syngeneic tumors revealed an 11-gene oxidative stress signature downregulated in Cbx3KO tumors after combined treatment; its low expression score correlated with improved overall survival in CRC patients. Among them, Slc7a11, Ptgs2, and Cth were identified as CBX3 direct target genes. Additionally, several amino acid transporters functionally coupled to Xc- antiporter, as Slc3a2, Slc7a5, Slc1a5 and Slc6a9, all significantly decreased in Cbx3KO tumor under RSL3+IFN{gamma} treatment. Mechanistically, CBX3 binds directly to the promoter of Slc7a11 or Slc3a2 to sustain their expression and redox balance to resist ferroptosis in CRC. Knockout of CBX3 in CRC cells significantly impaired RNA polymerase II recruitment to the promoter of SLC7A11 or SLC3A2 and reduced SLC7A11 and SLC3A2 expression, an effect that was further exacerbated under IFN{gamma} stimulation but restored by STAT1 deletion. Overall, CBX3 protects CRC from oxidative stress by maintaining the functional integrity of IFN{gamma}/STAT1/SLC7A11/SLC3A2 axis, positioning CBX3 as a potential therapeutic target for enhancing ferroptosis therapies in CRC.

cancer biology↗

bioRxiv2025

Mucosal vaccine immunity induced by a new auxotrophic Pseudomonas aeruginosa strain is linked to Th17 and IgA responses

Pseudomonas aeruginosa (P.a) is a Gram-negative opportunistic pathogen that poses a major global health threat, particularly in immunocompromised individuals, patients with cystic fibrosis, and those with burn injuries or ventilator-associated pneumonia. Despite intense efforts, no licensed vaccine is currently available for human use. In this context, live attenuated vaccines (LAVs) represent a promising but underexplored approach, offering the potential to elicit robust, long-lasting, and multifaceted immune responses including that of inducing trained immunity. Here, we sub-cultured {Delta}LasB PAO1 (a P.a strain that we have shown previously shown to have reduced virulence) in artificial sputum medium (ASM), a culture medium mimicking CF sputum in which bacteria often show auxotrophy. We showed that such a strain (designed here V for vaccine) was auxotrophic, less virulent, and had characteristics of CF-like strains. Crucially, V was able to induce both local (IgA) and systemic humoral responses as well as memory Th17 immune responses, and could, when administered intra-tracheally (but not intra-muscularly), fully protected mice against a lethal PAO1 infection. Overall, the present study demonstrates that our vaccine formulation, in addition to providing an advantageous auxotrophic phenotype adapted to the CF setting, was efficient, when given mucosally, in preferentially inducing secretory IgA and Th17 pathway at mucosal surfaces, a critical barrier that neutralizes pathogens before tissue invasion.

immunology↗

bioRxiv2024

Signaling downstream of tumor-stroma interaction regulates mucinous colorectal adenocarcinoma apicobasal polarity

Mucinous colorectal carcinoma (MUC CRC) dissemination into the tumor stroma and metastasis to multiple organs, including the peritoneum, is associated with poor prognosis. Disseminating MUC CRCs exhibit either a conventional apical-in or an inverted apical-out polarity phenotype that influence patient outcome. Identifying the mechanisms controlling MUC CRC polarity is critical to understand disease progression. Here, we analyze patient-derived MUC CRC xenografts, with apical-in or apical-out polarity, ex vivo or within collagen gels to mimic the peritumoral stroma. Single-cell analyses reveal 2{beta}1-integrin as a key collagen-binding receptor in these models. Collagen-2{beta}1-integrin interaction activates Src and ERK/MAPK signaling and upregulates the expression of SorLA, an endosomal sorting receptor. SorLA supports apical-in polarity and carcinoma-stroma interactions by promoting integrin recycling to the plasma membrane and HER2/HER3 expression through a positive feedback mechanism. Accordingly, we observe positive correlation between HER2, HER3 and SorLA in patient samples with the highest HER2 expression in apical-in-presenting tissues. Treatment of tumor spheres with clinically relevant HER2/HER3-targeting antibodies reverts sphere polarity and impedes collagen remodeling and adhesion to mouse peritoneum. This SorLA--integrin--HER2/HER3 signaling axis may represent a basis for MUC CRC-patient stratification and shed light on other carcinomas with similar apical-out phenotypes.

cancer biology↗

bioRxiv2023

mTOR Inhibition Suppresses Salinomycin-Induced Ferroptosis in Breast Cancer Stem Cells by Ironing Out Mitochondrial Dysfunctions

Ferroptosis constitutes a promising therapeutic strategy against cancer by efficiently targeting the highly tumorigenic and treatment-resistant cancer stem cells (CSCs). We previously showed that the lysosomal iron-targeting drug Salinomycin (Sal) was able to eliminate CSCs by triggering ferroptosis. Here, in a well-established breast CSCs model (human mammary epithelial HMLER CD24low/CD44high), we identified that pharmacological inhibition of mechanistic target of rapamycin (mTOR), suppresses Sal-induced ferroptosis. Mechanistically, mTOR inhibition modulates iron cellular flux and prevents the iron and ROS bursts induced by Sal. Besides, integration of multi-omics data identified mitochondria as a key target of Sal action. We demonstrated that mTOR inhibition prevents Sal-induced mitochondrial functional and structural alteration, and that Sal-induced metabolic plasticity is mainly dependent on the mTOR pathway. Overall, our findings provide experimental evidences on the detailed mechanisms of mTOR as a crucial effector of Sal-induced ferroptosis, and gives proof-of-concept that careful evaluation of such combination therapy (here mTOR and ferroptosis co-targeting) is required for effective treatment. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=120 SRC="FIGDIR/small/539040v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@8b8772org.highwire.dtl.DTLVardef@695e0forg.highwire.dtl.DTLVardef@19e3f83org.highwire.dtl.DTLVardef@5bc0a6_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

bioRxiv2023

BCR-ABL promotes hematopoietic stem and progenitor cell formation in embryonic stem cells

Generating Hematopoietic Stem Cells (HSCs) from Pluripotent Stem Cells (PSCs) has been a long-lasting quest in the field of hematopoiesis. Previous studies suggested that enforced expression of BCR-ABL, the unique oncogenic driver of Chronic Myelogeneous Leukemia (CML), in Embryonic Stem Cells (ESCs)-derived hematopoietic cells is sufficient to confer long-term in vivo repopulating potential. To precisely uncover the molecular events regulated by the Tyrosine-kinase activity of BCR-ABL1 (p210) during the course of hematopoietic differentiation, we engineered a Tet-ON inducible system to modulate its expression in murine ESC. We showed in unique site-directed knock-in ESC model, that BCR-ABL expression tightly regulated by doxycycline (dox) controls the formation and the maintenance of immature hematopoietic progenitors. Interestingly, these progenitors can be expanded in vitro for several passages in the presence of dox. Our analysis of cell surface markers and transcriptome compared to wild-type fetal and adult HSCs unraveled a similar molecular signature. LTC-IC assay confirmed their self-renewal capacities albeit with a differentiation bias towards erythroid and myeloid cells. Collectively, our novel Tet-ON system represents a unique in vitro model to shed lights on ESC-derived hematopoiesis, CML initiation and maintenance. KEY POINTSO_LIWe report a unique BCR-ABL-induced-embryonic stem cell -derived hematopoiesis model in murine embryonic stem cells C_LIO_LIThis BCR-ABL-induced self-renewal and differentiation model can be of major interest to uncover molecular events required for ESC-derived hematopoiesis C_LI

cancer biology↗

bioRxiv2022

miR-495-3p Sensitizes BCR::ABL1 Expressing Leukemic cells to Tyrosine Kinase Inhibitors by Targeting Multidrug Resistance 1 Gene including in T315I Mutated cells

Chronic myeloid leukemia (CML) is a clonal hematopoietic malignancy driven by the BCR::ABL1 fusion oncoprotein. The development of tyrosine kinase inhibitors (TKIs) has deeply increased long-term survival of CML patients. Nonetheless, one patient out of four will switch TKI off owing either to drug intolerance or resistance partly due to amplification or mutations of BCR::ABL1 oncogene and alteration of ATP-binding cassette (ABC) transporters. Increasing evidence suggests an involvement of the microRNA miR-495-3p in cancer-associated chemo-resistance through multidrug resistance 1 (MDR1) gene which encodes an ATP-dependent efflux pump. Our study aimed at investigating the potential role of miR-495-3p in CML TKI chemo-sensitivity and determining the underlying molecular circuitry involved. We first observed that miR-495-3p expression was lower in BCR::ABL1 expressing cellular models in vitro. Notably, loss-of-function experiments showed increased proliferation associated with a decreased number of non-dividing cells (G0/G1) and resistance to Imatinib. Conversely, our data showed that miR-495-3p overexpression hindered leukemic cell growth and TKI resistance even in Imatinib-resistant T315I-mutant cells as well as drug efflux activity through MDR1 regulation. To further investigate the role of miR-495-3p in CML patients, we found that predicted miR-495-3p targets were upregulated in patients in blast crisis involved in protein phosphorylation and associated with the worst prognosis. Taken together, our results demonstrate that down-regulation of miR-495-3p expression is important in the malignant phenotype of CML and TKI resistance mechanisms, which could be a useful biomarker and a potential therapeutic target to eradicate CML. MeSH termsO_LIATP Binding Cassette Transporter, Subfamily B C_LIO_LIATP Binding Cassette Transporter, Subfamily B, Member 1 / drug effects C_LIO_LIATP Binding Cassette Transporter, Subfamily B, Member 1 / metabolism* C_LIO_LIBlast Crisis / pathology C_LIO_LICell Line, Tumor C_LIO_LICell Proliferation / drug effects* C_LIO_LICell Survival / drug effects C_LIO_LIDrug Resistance C_LIO_LIGenes, MDR C_LIO_LIImatinib Mesylate C_LIO_LILeukemia, Myelogenous, Chronic, BCR-ABL Positive C_LIO_LIMicroRNAs / genetics C_LIO_LIMicroRNAs / physiology* C_LI HIGHLIGHTSO_LImiR-495-3p inhibits leukemic cell growth and is downregulated in BCR::ABL1 expressing cell lines C_LIO_LImiR-495-3p modulates response to TKI treatment including in UT7 cells expressing T315I C_LIO_LIOverexpression of miR-495-3p leads to a decrease of MDR1 and drug efflux activity C_LIO_LIBioinformatics analyses reveal that MiR-495-3p target genes are upregulated in blast crisis C_LI

cancer biology↗

bioRxiv2022

Modeling Global Genomic Instability in Chronic Myeloid Leukemia (CML) using patient-derived induced pluripotent stem cells (iPSC)

Using induced Pluripotent Stem Cell (iPSC) technology, it is now possible to reprogram the primary leukemic cells to pluripotency and generate a major source of stem cells. To determine the feasibility of modeling global genomic instability characterizing chronic myeloid leukemia (CML) progression towards blast crisis (BC), we have used a patient-specific iPSC line that has been submitted to in vitro mutagenesis with the mutagenic agent N-ethyl-N-nitrosourea (ENU). Increased genomic instability was validated using g-H2AX and micronuclei assays. As compared to non-mutagenized cultures, these iPSC generated an increased numbers of progenitors (x 5-Fold) which proliferated in liquid cultures with a blast cell morphology. CGH array analyses were performed using ENU-treated iPSC-derived hematopoietic cells in two different time points as compared to leukemic iPSC cultured without ENU. Several cancer genes were found to be involved in the ENU-treated cultures, some known to be altered in leukemia (BLM, IKZF1, NCOA2, ALK, EP300, ERG, MKL1, PHF6 and TET1). Remarkably, transcriptome geodataset GSE4170 (Radich et al., 2006) allowed us to associate 125 of 249 of the aberrations that we detected in CML iPSC, with genes already described during progression of CML to BC (p-value =9.43exp32, after ANOVA with 1000 permutations). 38 most predictive aberrations allowed perfect reclassification of BC and chronic phase samples by unsupervised classification. Among these candidates, eleven of them have been described in CML physiopathology and connected to tyrosine kinase inhibitor (TKI) resistance and genomic instability. Overall, these results demonstrated that we have generated for the first time to our knowledge, an in vitro genetic instability model in CML, reproducing genomic events described in patients with BC. This tool using patient-derived iPSC could be of interest to study CML progression, to discover novel targets and eventually to test novel therapies.

cancer biology↗

bioRxiv2021

NR0B2 regulation during Primary Sclerosing Cholangitis defines a metabolic and pre-malignant reprogramming of Cholangiocyte

Primary Sclerosing Cholangitis (PSC) is an idiopathic, cholestatic liver disease that is characterized by persistent, progressive, biliary inflammation leading to cirrhosis. These patients present higher risk for developing bile duct cancers. Biomedical text-mining related to PSC symptoms like: biliary inflammation, biliary fibrosis, biliary cholestasis was initiated to collect gene associations with this pathophysiology. The text mining work was integrated in distinct omics data such as human transcriptome of PSC liver, Farnesoid X receptor (FXR) functional liver transcriptome and liver single cell transcriptome of the Abcb4-/- model of PSC. A molecular network implicated in abnormal hepatobiliary system physiology was built and confirming a major implication of Nr0b2 and its associated nuclear receptors like FXR in a metabolic cascade that could influence immune response. TNFRSF12A/TWEAK receptor, was found up regulated in PSC liver independently of FXR regulation and TWEAK signaling is known for its implication in pre-conditioning niche of cholangiocarcinoma. NR0B2 deregulation in PSC liver was found independent of gender, age and body mass index surrogates. At single cell level, Nr0b2 up regulation was found in cholangiocytes but not in hepatocytes. In affected cholangiocytes, the cell trajectory built on Nr0b2 expression, revealed implication of several metabolic pathways for detoxification like sulfur, glutathione derivative and monocarboxylic acid metabolisms. On this cell trajectory it was discovered some molecules potentially implicated in carcinogenesis like: GSTA3, ID2 and mainly TMEM45A a transmembrane molecule from golgi apparatus considered as oncogene in several cancers. All together, these observations found in humanPSC liver and in its murine models allowed to highlight an independent deregulation of NR0B2 with a metabolic and premalignant reprogramming of cholangiocytes.

pathology↗

bioRxiv2021

ENOX2 NADH Oxidase: A BCR-ABL1-dependent cell surface and secreted redox protein in chronic myeloid leukemia (CML)

Chronic myeloid leukemia (CML) is a myeloproliferative neoplasm caused by the acquisition of BCR-ABL1 fusion in a hematopoietic stem cell. We identified the ENOX2 gene as up-regulated in BCR-ABL1-expressing UT-7 cell lines through a transcriptome assay. The oncofoetal ENOX2 protein (Ecto-Nicotinamide Adenine Dinucleotide Oxidase Disulfide Thiol Exchanger 2) is expressed on the external plasma membrane surface of cancer cells and can be released in cancer patients serum. Considering these data, we studied ENOX2 expression in CML cell lines and patients using quantitative RT-PCR, western-blots, the ELISA method, and transcriptomic dataset reanalysis. We confirmed increased ENOX2 mRNA expression in the BCR-ABL1-expressing UT-7 cell line. Comparable results were obtained in CML patients at diagnosis. Western-blot analyses on UT-7 and TET-inducible Ba/F3 cell lines established the up-regulation of ENOX2 protein. BCR-ABL1 has been found to induce ENOX2 overexpression in a kinase-dependent manner. In a series of 41 patients with CML, ELISA assays showed a highly significant increase of ENOX2 protein levels in the plasma of patients with CML (p < 0.0001) as compared to controls (n=28). Transcriptomic dataset (GSE4170) reanalyzes have shown specific ENOX2 mRNA overexpression in the chronic phase of the disease. Bioinformatic analyses identified several genes whose mRNA expression was positively correlated to ENOX2. Some of them encode proteins involved in cellular functions compatible with the growth deregulation observed in CML. All in all, our results demonstrate for the first time the upregulation of a secreted Redox protein in a BCR-ABL1-dependent manner in CML. Our data suggest that ENOX2 (through its transcriptional program) plays a significant role in the BCR-ABL1 leukemogenesis. Further studies are required to clarify the relationship between BCR-ABL1 and ENOX2.

cancer biology↗

bioRxiv2021

Detection of hematopoietic stem cell transcriptome in human fetal kidneys and kidney organoids derived from human induced pluripotent stem cells (iPSC)

BackgroundIn mammalians, hematopoietic stem cells (HSC) arise in the dorsal aorta from the hemogenic endothelium, followed by their migration to fetal liver and to bone marrow. In zebrafish, kidney is the site of primary hematopoiesis. In humans, the presence of HSC in the fetal or adult kidney has not been established. MethodsWe analyzed the presence of HSC markers in human fetal kidneys by analysis of single-cell datasets. We then analyzed in kidney organoids derived from iPSC, the presence of hematopoietic markers using transcriptome analyses. Results12 clusters were identified of stromal, endothelial, and nephron cell type-specific markers in the two fetal stage (17 weeks) kidney datasets. Among these, expression of hematopoietic cells in Cluster 9 showed expression of primitive markers. Moreover, whole transcriptome analysis of our iPSC-derived kidney organoids revealed induction of the primitive hematopoietic transcription factor RUNX1 as found in the human fetal kidney cortex. ConclusionsThese finding support the presence of cells expressing HSC transcriptome in human kidney. The mechanisms of the appearance of the cells with the same transcriptional features during iPSC-derived kidney organoid generation requires further investigation.

developmental biology↗

bioRxiv2020

Direct reprogramming of adult hepatocytes to generate LGR5+ endodermal progenitor

We successfully converted hepatocytes isolated from adult mice into expandable and stable leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5)-positive endodermal progenitor cells (EndoPCs). This was accomplished in vitro through transient exposure to four transcriptional factors (OCT3/4, SOX2, KLF4, and cMYC) and STAT3 activators. EndoPCs were generated by a process that involved an epithelial-mesenchymal transition (EMT) without the use of any components of the canonical WNT/{beta}-catenin or LGR5/R-spondin signaling pathways. We showed that the proliferation and capacity for self-renewal of EndoPCs in 2D long-term culture were controlled by three interrelated signaling pathways: gp130/JAK/STAT3, LGR5/R-spondin, and WNT/{beta}-catenin. After long-term maintenance in two- and three-dimensional culture systems, EndoPCs were able to differentiate into liver-restricted lineages such as hepatocyte-like cells and bile duct-like structures in vitro and in vivo. After intra-muscular injection, EndoPCs generated macroscopically visible and well-vascularized liver-like tissue, which contained Alb+ liver parenchyma-like structures and substantial, KRT7/KRT19+ bile duct-like cell organizations. ConclusionWe have developed an efficient method for producing LGR5+ adult endodermal stem cells. These cells will be useful for the in vitro study of the molecular mechanisms of liver development and have important potential for therapeutic strategies, including approaches based on bioengineered liver tissue. These cells also open up new avenues for experiments focused on disease modeling, toxicology studies, and regenerative medicine applications.

developmental biology↗

bioRxiv2020

Pharmacologically modified pluripotent stem cell-based cancer vaccines with anti-metastatic potential

Cancer is maintained by the activity of a rare population of self-renewing "cancer stem cells" (CSCs), which are resistant to conventional therapies. CSCs share several antigenic determinants with pluripotent stem cells (PSCs). We show here that PSCs, combined with a histone deacetylase inhibitor (HDACi), are able to elicit major anti-tumor responses in a model of highly aggressive breast cancer. This immunotherapy strategy was effective in preventing tumor establishment and efficiently targeted CSCs by inducing extensive modifications of the tumor microenvironment. The anti-tumor effect was correlated with a reduction in regulatory T and myeloid-derived suppressor cell populations and an increase in cytotoxic CD8+T cells within the tumor and the spleen along with a drastic reduction in metastatic dissemination and an improvement in the survival rate. These results demonstrate for the first time the possibility of using PSCs and HDACi as an allogeneic anticancer vaccine, in future universal immunotherapy strategies.

cancer biology↗

bioRxiv2019

Evidence of massive leukemic progenitor expansion in blood during molecular recurrence after TKI discontinuation in chronic myeloid leukemia (CML)

Chronic myeloid leukemia (CML) represents one of the major success stories of targeted therapies of the 21st century with the use of tyrosine kinase inhibitors (TKIs). Following discontinuation of TKIs in deep molecular remission context, at least half of the patients experience molecular relapse. Cellular events occurring during this phase are not yet established. In this work, we show a massive amplification of clonogenic progenitors or CFCs (colony-forming cells) in the peripheral blood of patients in molecular recurrence after a TKI discontinuation. We demonstrate, by qRT-PCR analysis on individual and pooled CFCs, that leukemic clonogenic progenitor expansion in the peripheral blood represents the first cellular event before any evidence of cytological change. These findings also suggest that the amplification of the leukemic clone during the first stages of CML takes place not only in the bone marrow but also in the peripheral blood.

cancer biology↗

bioRxiv2019

iPSC-derived Cancer Organoids Recapitulate Genomic and Phenotypic Alterations of c-met-mutated Hereditary Kidney Cancer

Hereditary cancers with cancer-predisposing mutations represent unique models of human oncogenesis as a driving oncogenic event is present in germline, exposing the healthy member of a family to the occurrence of cancer. The study of the secondary events in a tissue-specific manner is now possible by the induced pluripotent stem cell (iPSC) technology offering the possibility to generate an unlimited source of cells that can be induced to differentiate towards a tissue at risk of malignant transformation. We report here for the first time, the generation of a c-met-mutated iPSC lines from the somatic cells of a patient with type 1 papillary renal cell carcinoma (PRCC). We demonstrate the feasibility of kidney differentiation with iPSC-derived organoids expressing markers of kidney progenitors with presence of tight junctions and brush borders in tubular structures at transmission electron microscopy. Importantly, c-met-mutated kidney organoids expressed PRCC markers both in vitro and in vivo in NSG mice. Gene expression profiling of c-met-mutated iPSC-derived organoid structures showed striking molecular similarities with signatures found in a large cohort of PRCC patient samples and identified 11 common genes. Among these, BHLHE40 and KDM4C, well-known factors involved in PRCC pathogenesis, were expressed in c-met-mutated kidney organoids. This analysis applied to primary cancers with and without c-met mutation showed overexpression of the BHLHE40 and KDM4C only in the c-met-mutated PRCC tumors, as predicted by c-met-mutated organoid transcriptome. These data represent therefore the first proof of concept of the generation of \"renal carcinoma in a dish\" model using c-met-mutated iPSC-derived organoids, opening new perspectives for discovery of novel potentially predictive disease markers and novel drugs for future precision medicine strategies.

cancer biology↗

bioRxiv2017

Transgene-free hematopoietic stem and progenitor cells from human induced pluripotent stem cells

Introductory paragraphThe successful production of Hematopoietic Stem and Progenitor Cells (HSPCs) from human pluripotent sources is conditioned by transgene delivery1-5. We describe here a dedicated and tractable one step, GMP-grade, transgene-free and stroma-free protocol to produce HSPCs from human induced pluripotent stem cells (hiPSCs). This procedure, applied to several sources of hiPSCs with equal efficiency, is based on a directed differentiation with morphogens and cytokines to generate a cell population close to nascent HSPCs or their immediate forerunners i.e., hemogenic endothelial cells6-9. Following engraftment into immunocompromised recipients, this cell population was proved capable of a robust myeloid, lymphoid and definitive red blood cell production in sequential recipients for at least 40 weeks. Further identification of the repopulating cells show that they express the G protein-coupled receptor APELIN (APLNR) and the homing receptor CXCR4. This demonstrates that the generation of bona fide HSPCs from hiPSCs without transgenes is possible and passes through an early endo-hematopoietic intermediate. This work opens the way to the generation of clinical grade HSPCs for the treatment of hematological diseases and holds promise for the analysis of HSPC development in the human species.

cell biology↗