Biology subjects

Hovey, O. F. J.

Publications and source records attributed to Hovey, O. F. J..

3 recordsLinked to original sources

bioRxiv2026

Pharmacological inhibition of LIN28A promotes imatinib sensitivity in CML resistance

Resistance to tyrosine kinase inhibitors (TKIs) remains a critical challenge in chronic myeloid leukemia (CML), particularly when driven by mechanisms independent of BCR-ABL1 kinase-domain mutations. Building on the identification of the RNA-binding protein LIN28A as a driver of imatinib resistance, we evaluated emerging LIN28 inhibitors as potential sensitizing agents. Screening three small molecules in an imatinib-resistant (ImR) K562 model identified LIN28i-1632 as uniquely synergistic with imatinib (synergy score: 12.07), reducing cell proliferation by 71.15%. Quantitative DIA and TMT proteomics revealed that this synergy is characterized by significant proteomic remodelling, including the downregulation of the canonical LIN28 target HMGA1 and the activation of apoptotic and G2/M cell cycle checkpoint programs. Mechanistically, phosphoproteome and kinome profiling showed suppressed AKT/RPS6K and CDK signalling. We further demonstrate that LIN28i-1632 promotes sensitivity by reducing BCR-ABL protein abundance and attenuating the AKT survival axis through RICTOR downregulation and PTEN restoration. Collectively, our findings establish pharmacological LIN28 inhibition as a viable strategy to overcome TKI resistance by simultaneously engaging cell-cycle arrest and dismantling the AKT-mediated survival network.

cancer biology↗

bioRxiv2026

Proteomic profiling reveals pleiotropic antimetabolite activity of triciribine in acute lymphoblastic leukemia

Acute lymphoblastic leukemia (ALL) exhibits marked genetic and metabolic heterogeneity that limits the efficacy of targeted therapies. Antimetabolite strategies remain central to ALL treatment, yet the mechanisms underlying differential drug sensitivity are incompletely defined. Here, we investigated the activity of the purine analog triciribine (TCN) across diverse ALL cellular models. We find that TCN exerts potent cytotoxic effects in multiple ALL cell lines, exceeding those observed with canonical Akt inhibitors in our datasets. Phosphoproteomic analyses indicate that, at early time points, TCN does not primarily suppress Akt signaling but instead induces transient pathway activation accompanied by inhibition of cyclin-dependent kinases in both sensitive and resistant cells. The monophosphorylated metabolite TCN-P represents the predominant intracellular species and requires adenosine kinase (ADK) for activity, with ADK protein levels positively correlating with TCN sensitivity in both cell lines and primary patient samples. Using Proteome Integral Solubility Alteration profiling, we identify candidate protein interactions of TCN-P distributed across multiple cellular pathways, including nucleotide metabolism, DNA replication, and protein synthesis. These interactions are accompanied by impaired purine biosynthesis, DNA damage, translational stress, and cell-cycle arrest, as supported by time-course quantitative proteomics and immunoblot analyses. Together, these findings characterize triciribine as an antileukemic agent with pleiotropic antimetabolite activity in ALL and highlight ADK-dependent metabolism as a key determinant of therapeutic sensitivity, suggesting that ADK levels may serve as a predictive biomarker to stratify patients for triciribine-based precision treatment strategies.

cancer biology↗

bioRxiv2025

Isolation of Extracellular Vesicles from Minimal Volume Ascites Fluid Using Strong Anion Exchange Magnetic Beads

Ovarian cancer (OC) remains a leading cause of gynecologic cancer mortality due to late-stage diagnosis and limited early detection strategies. Ascites fluid, a pathological hallmark of OC, is a rich source of tumor-derived extracellular vesicles (EVs) that reflect the tumor microenvironment and hold promise for biomarker discovery. However, isolating EVs from minimal ascites volumes (<100 {micro}L) poses technical challenges using conventional methods like ultracentrifugation or size-exclusion chromatography (SEC). This study explores the application of strong anion exchange (SAX) magnetic beads (Mag-Net) for efficient EV isolation from as little as 2 {micro}L of ascites fluid from both murine models and a human patient with mucinous borderline tumor. We demonstrate that SAX achieves robust EV capture at 10{micro}l of input volume, enabling comprehensive proteomic profiling and single-EV surface-enhanced Raman spectroscopy (SERS) with a >2-fold increase in proteomic depth compared to raw ascites. Notably, this study was able to identify 1000 proteins not previously annotated in Vesiclepedia for OC-derived EVs, alongside distinct SERS signatures, highlighting the potential for multiomic analysis. Comparative analysis with UC revealed enhanced proteomic depth obtained with SAX beads, albeit we also observed differential detection of canonical markers (e.g., CD9, CD81) between input volumes of ascites fluid. These findings establish SAX as a scalable, low-input platform for EV-based biomarker discovery, paving the way for improved early detection and molecular insights into OC progression.

biochemistry↗