Biology subjects

Blind, R.

Publications and source records attributed to Blind, R..

5 recordsLinked to original sources

bioRxiv2026

Host soluble inositol phosphate signaling promotes coronavirus replication

Coronaviruses rely extensively on host pathways for replication, making host-directed therapies an attractive strategy for broad-spectrum antivirals with reduced risk of viral resistance. Here we identify the host soluble inositol phosphate pathway as a previously unrecognized dependency for coronavirus infection. Genetic or pharmacologic inhibition of several kinases in this pathway markedly suppresses replication of both alpha- and betacoronaviruses, while increasing pathway activity promotes viral replication. We developed UNC7844, a potent multi-target inhibitor of these kinases, which reduces coronavirus replication by more than four orders of magnitude in cultured cells and suppresses coronavirus infection in mice. Mechanistically, UNC7844 suppresses inositol (pyro)phosphates production, disrupts phosphoinositide homeostasis, and impairs late endosomal dynamics, blocking early post-entry steps required for viral genome release and replication. Together, our findings establish the soluble inositol (pyro)phosphate pathway as an important regulator of coronavirus infection and highlight its inhibition as a promising host-directed antiviral strategy.

microbiology↗

bioRxiv2026

Structure-guided compound prioritization strategy for virtual screening identifies putative binders for the nuclear receptor LRH-1

Compound ranking in structure-based virtual screening notoriously yields highly ranked false positive binders due to variable poses or biases in scoring terms. We developed a compound prioritization strategy that utilizes sampled docked poses from contrasting docking approaches (targeted physics-based docking and blind docking with a generative model) against multiple models of the target protein to train a multi-layer perceptron (MLP). The model predicts binders at the orthosteric ligand-binding pocket of the nuclear receptor LRH-1 (NR5A2). Our approach circumvents the reliance on a single docked pose for scoring compounds or individual scoring metrics for compound ranking. In a separate benchmarking set, we observed that the MLP identifies known binders that are chemically dissimilar from the compounds in the training set and is sensitive to single scaffold modifications, making it a potential tool for lead optimization. We applied our strategy to a prospective virtual screening campaign, which resulted in the discovery of four putative LRH-1 binders. We found that a combination of scoring and prediction metrics enriches for the hit compounds across library sizes. In all, this implementation presents a method to leverage structural and experimental data to aid virtual screening for a challenging protein target.

bioinformatics↗

bioRxiv2024

A novel heuristic of rigid docking scores positively correlates with full-length nuclear receptor LRH-1 regulation.

The nuclear receptor Liver Receptor Homolog-1 (LRH-1, NR5A2) is a ligand-regulated transcription factor and validated drug target for several human diseases. LRH-1 activation is regulated by small molecule ligands, which bind to the ligand binding domain (LBD) within the full-length LRH-1. We recently identified 57 compounds that bind LRH-1, and unexpectedly found these compounds regulated either the isolated LBD, or the full-length LRH-1 in cells, with little overlap. Here, we correlated compound binding energy from a single rigid-body scoring function with full-length LRH-1 activity in cells. Although docking scores of the 57 hit compounds did not correlate with LRH-1 regulation in wet lab assays, a subset of the compounds had large differences in binding energy docked to the isolated LBD vs. full-length LRH-1, which we used to empirically derive a new metric of the docking scores we call "{Delta}{Delta}G". Initial regressions, correlations and contingency analyses all suggest compounds with high {Delta}{Delta}G values more frequently regulated LRH-1 in wet lab assays. We then docked all 57 compounds to 18 crystal structures of LRH-1 to obtain averaged {Delta}{Delta}G values, which robustly and reproducibly associated with full-length LRH-1 activity in cells. Network analyses on the 18 crystal structures of LRH-1 suggest unique communication paths exist between the subsets of LRH-1 crystal structures that produced high vs. low {Delta}{Delta}G values, identifying a structural relationship between {Delta}{Delta}G and the position of Helix 6, a previously established regulatory helix important for LRH-1 regulation. Together, these data suggest computational docking can be used to quickly calculate {Delta}{Delta}G, which positively correlated with the ability of these 57 hit compounds to regulate full-length LRH-1 in cell-based assays. We propose {Delta}{Delta}G as a novel computational tool that can be applied to LRH-1 drug screens to prioritize compounds for secondary screening. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=115 SRC="FIGDIR/small/592617v1_ufig1.gif" ALT="Figure 1"> View larger version (41K): org.highwire.dtl.DTLVardef@1e3a780org.highwire.dtl.DTLVardef@109fcdcorg.highwire.dtl.DTLVardef@b80fa6org.highwire.dtl.DTLVardef@b68fe_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGraphical Abstract:C_FLOATNO C_FIG

pharmacology and toxicology↗

bioRxiv2024

Bilirubin is a new ligand for nuclear receptor Liver Receptor Homolog-1.

The nuclear receptor Liver Receptor Homolog-1 (LRH-1, NR5A2) binds to phospholipids that regulate important LRH-1 functions in the liver. A recent compound screen unexpectedly identified bilirubin, the product of liver heme metabolism, as a possible ligand for LRH-1. Here, we show unconjugated bilirubin directly binds LRH-1 with apparent Kd=9.3uM, altering LRH-1 interaction with all transcriptional coregulator peptides tested. Bilirubin decreased LRH-1 protease sensitivity, consistent with MD simulations predicting bilirubin stably binds LRH-1 within the canonical ligand binding site. Bilirubin activated a luciferase reporter specific for LRH-1, dependent on co-expression with the bilirubin membrane transporter SLCO1B1, but bilirubin failed to activate ligand-binding genetic mutants of LRH-1. Gene profiling in HepG2 cells shows bilirubin selectively regulated transcripts from endogenous LRH-1 ChIP-seq target genes, which was significantly attenuated by either genetic knockdown of LRH-1, or by a specific chemical competitor of LRH-1. Gene set enrichment suggests bilirubin and LRH-1 share roles in cholesterol metabolism and lipid efflux, thus we propose a new role for LRH-1 in directly sensing intracellular levels of bilirubin.

biochemistry↗

bioRxiv2024

Multiple inositol phosphate species enhance stability of active mTOR

Mechanistic Target of Rapamycin (mTOR) binds the small metabolite inositol hexakisphosphate (IP6) as shown in structures of mTOR, however it remains unclear if IP6, or any other inositol phosphate species, can activate mTOR kinase activity. Here, we show that multiple, exogenously added inositol phosphate species (IP6, IP5, IP4 and IP3) can all enhance the ability of mTOR and mTORC1 to auto-phosphorylate and incorporate radiolabeled phosphate into peptide substrates in in vitro kinase reactions. Although IP6 did not affect the apparent KM of mTORC1 for ATP, monitoring kinase activity over longer reaction times showed increased product formation, suggesting inositol phosphates stabilize an active form of mTORC1 in vitro. The effects of IP6 on mTOR were reversible, suggesting IP6 bound to mTOR can be exchanged dynamically with the free solvent. Interestingly, we also observed that IP6 could alter mTOR solubility and electrophoretic mobility in SDS-PAGE in the presence of manganese, suggesting divalent cations may play a role in inositol phosphate regulation of mTOR. Together, these data suggest for the first time that multiple inositol phosphate species (IP4, IP5 and IP6) can dynamically regulate mTOR and mTORC1 by promoting a stable, active state of the kinase. Our data suggest that studies of the dynamics of inositol phosphate regulation of mTOR are well justified.

biochemistry↗