Biology subjects

Leslie, K. K.

Publications and source records attributed to Leslie, K. K..

5 recordsLinked to original sources

bioRxiv2026

Auranofin potentiates cisplatin response through context-dependent NOTCH-associated signaling states in endometrial cancer

Therapeutic resistance remains a major challenge in advanced and recurrent endometrial cancer (EC). Aberrant NOTCH signaling has been associated with aggressive tumor behavior and therapeutic resistance across multiple malignancies, yet its therapeutic significance in EC remains incompletely defined. We investigated whether auranofin (AuR), a noncanonical modulator of NOTCH signaling through the transcriptional effector RBPJ, alters platinum responsiveness in EC models. Elevated NOTCH3 copy-number was associated with poorer overall survival in the TCGA-UCEC cohort. AuR treatment reduced RBPJ occupancy at canonical NOTCH target loci, including HES1 and HES4, across multiple EC models. Stable NOTCH3 depletion altered AuR responsiveness in a context-dependent manner while significantly enhancing cisplatin (CDDP) sensitivity in AN3CA cells. Pharmacologic AuR treatment similarly potentiated CDDP response in AN3CA cells and AN3CA xenografts, resulting in reduced tumor burden and prolonged endpoint-free survival following combination treatment. In contrast, ARK-1 xenografts demonstrated limited additional benefit from combined AuR plus CDDP therapy despite detectable suppression of RBPJ occupancy. Together, these findings identify context-dependent NOTCH-associated therapeutic vulnerabilities in EC and support further development of biomarker-guided AuR-based platinum-sensitization strategies. Statement of significanceAuranofin suppresses RBPJ-associated transcriptional activity and enhances cisplatin response in biologically distinct subsets of endometrial cancer.

cancer biology↗

bioRxiv2025

Curcumin Analogues Trigger HMOX1-Mediated Ferroptosis to Halt Endometrial Cancer Growth

BackgroundAdvanced endometrial cancer remains challenging to treat due to limited therapeutic options and drug resistance. Ferroptosis, an iron-dependent form of cell death, offers a potential strategy for overcoming resistance. Curcumin analogues with improved bioavailability, such as HO-3867 and AKT-100, exhibit potent anti-cancer activity, but their mechanisms remain under-explored. MethodsKLE, Hec50co and Ishikawa endometrial cancer cells were treated with HO-3867 or AKT-100. RNA sequencing, qPCR, and immunoblotting assessed ferroptosis-related gene expression, focusing on HMOX1. Intracellular iron and ROS were measured via FerroOrange and DCFH-DA staining. Cytotoxicity and colony formation were evaluated using CyQUANT assays, with pharmacological inhibitors (ZnPP, Liproxstatin-1, Z-VAD-FMK) and HMOX1 siRNA to dissect the roles of ferroptosis and apoptosis. ResultsBoth analogues upregulated multiple ferroptosis genes, prominently HMOX1. AKT-100 increased intracellular iron and ROS levels. Inhibition of HMOX1, ferroptosis, or apoptosis partially rescued cell viability, while HMOX1 knockdown enhanced clonogenic growth, confirming its key role in AKT-100-mediated cytotoxicity. ConclusionsHO-3867 and AKT-100 induce HMOX1-mediated ferroptosis and apoptosis, effectively suppressing endometrial cancer cell growth. These findings support the therapeutic potential of curcumin analogues and provide a foundation for in vivo studies targeting HMOX1 in endometrial cancer. O_FIG O_LINKSMALLFIG WIDTH=187 HEIGHT=200 SRC="FIGDIR/small/689080v1_ufig1.gif" ALT="Figure 1"> View larger version (37K): org.highwire.dtl.DTLVardef@610bcorg.highwire.dtl.DTLVardef@34d8eaorg.highwire.dtl.DTLVardef@18d2a5aorg.highwire.dtl.DTLVardef@d7b4f3_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

bioRxiv2025

The novel curcumin analogue AKT-100 targets mutant p53 in gynecologic cancer cells

Loss of normal p53 tumor-suppressive activity is common in advanced cancers, where TP53 mutations produce either p53-null states or highly expressed missense proteins with gain of oncogenic function. Mutant p53 proteins not only lose wild type p53 pro-apoptotic and DNA repair functions but also acquire new activities, including constitutive STAT3 activation. We hypothesize that directly targeting missense mutant p53 instead of downstream pathways is advantageous. Curcumin analogues have been reported to bind mutant p53, motivating the development of AKT-100, a novel analogue designed to restore wild type p53 functionality. AKT-100 binds mutant p53 and STAT3, as shown by fluorescence quenching, and inhibits STAT3 Tyr705 phosphorylation in KLE and COV362 cells. This agent exhibits potent cytotoxicity in gynecological cancer cells with IC50 values ranging from 4 nm to 2.51 {micro}M, and demonstrates synergy with the PARP inhibitor olaparib and with chemotherapy by suppressing alternative DNA repair mechanisms associated with therapeutic resistance. RNA sequencing and immunoblotting confirms AKT-100 reactivates wild type p53 transcriptional profiles regulating cell cycle arrest (CDKN1A/p21, GADD45A), apoptosis (PMAIP1/Noxa, DR5), and inhibits DNA replication and repair. These findings support AKT-100 as a promising therapeutic agent with the dual capabilities of directly targeting oncogenic mutant p53 and constitutively active STAT3. HighlightsO_LIAKT-100 is a novel curcumin analogue that functions as a WT p53 reactivator. C_LIO_LIAKT-100 inhibits STAT3 phosphorylation at Tyrosine 705 in endometrial and ovarian cancer cells. C_LIO_LICell proliferation is inhibited in multiple ovarian and serous endometrial cancer cell lines with AKT-100 as a single agent. C_LIO_LIAKT-100 is synergistic with PARP inhibitor olaparib and chemotherapy drugs carboplatin and paclitaxel. C_LI Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=132 SRC="FIGDIR/small/680794v3_ufig1.gif" ALT="Figure 1"> View larger version (48K): org.highwire.dtl.DTLVardef@1fdb8a8org.highwire.dtl.DTLVardef@6648acorg.highwire.dtl.DTLVardef@12d301aorg.highwire.dtl.DTLVardef@1b1370f_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

bioRxiv2023

Multiplexed live-cell imaging for drug responses in patient-derived organoid models of cancer

Patient-derived organoid (PDO) models of cancer are a multifunctional research system that better recapitulates human disease as compared to cancer cell lines. PDO models can be generated by culturing patient tumor cells in extracellular basement membrane extracts (BME) and plating as three-dimensional domes. However, commercially available reagents that have been optimized for phenotypic assays in monolayer cultures often are not compatible with BME. Herein we describe a method to plate PDO models and assess drug effects using an automated live-cell imaging system. In addition, we apply fluorescent dyes that are compatible with kinetic measurements to simultaneously quantitate cell health and apoptosis. Image capture can be customized to occur at regular time intervals over several days. Users can analyze drug effects in individual Z-plane images or a Z Projection of serial images from multiple focal planes. Using masking, specific parameters of interest are calculated, such as PDO number, area, and fluorescence intensity. We provide proof-of-concept data demonstrating the effect of cytotoxic agents on cell health, apoptosis and viability. This automated kinetic imaging platform can be expanded to other phenotypic readouts to understand diverse therapeutic effects in PDO models of cancer. SUMMARYPatient-derived tumor organoids are a sophisticated model system for basic and translational research. This methods article details the use of multiplexed fluorescent live-cell imaging for simultaneous kinetic assessment of different organoid phenotypes.

cancer biology↗

bioRxiv2022

Characterization of a novel estrogen- and progesterone-responsive endometrial cancer cell line: HCI-EC-23

Most endometrial cancers express the hormone receptor estrogen receptor alpha (ER) and are driven by excess estrogen signaling. However, evaluation of the estrogen response in endometrial cancer cells has been limited by the availability of hormonally responsive in vitro models, with one cell line, Ishikawa, being used in most studies. Here, we describe a novel, adherent endometrioid endometrial cancer (EEC) cell line model, HCI-EC-23. We show that HCI-EC-23 retains ER expression and that ER functionally responds to estrogen induction over a range of passages. We also demonstrate that this cell line retains paradoxical activation of ER by tamoxifen, which is also observed in Ishikawa and is consistent with clinical data. The mutational landscape shows that HCI-EC-23 is mutated at many of the commonly altered genes in EEC, has relatively few copy-number alterations, and is microsatellite instable high (MSI-high). In vitro proliferation of HCI-EC-23 is strongly reduced upon combination estrogen and progesterone treatment. HCI-EC-23 exhibits strong estrogen dependence for tumor growth in vivo and tumor size is reduced by combination estrogen and progesterone treatment. Molecular characterization of estrogen induction in HCI-EC-23 revealed hundreds of estrogen-responsive genes that significantly overlapped with those regulated in Ishikawa. Analysis of ER genome binding identified similar patterns in HCI-EC-23 and Ishikawa, although ER exhibited more bound sites in Ishikawa. This study demonstrates that HCI-EC-23 is an estrogen- and progesterone-responsive cell line model that can be used to study the hormonal aspects of endometrial cancer.

cancer biology↗