<?xml version="1.0" encoding="UTF-8" ?>
<rdf:RDF xmlns:admin="http://webns.net/mvcb/" xmlns="http://purl.org/rss/1.0/" xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:prism="http://purl.org/rss/1.0/modules/prism/" xmlns:taxo="http://purl.org/rss/1.0/modules/taxonomy/" xmlns:content="http://purl.org/rss/1.0/modules/content/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:syn="http://purl.org/rss/1.0/modules/syndication/">
<channel rdf:about="https://biorxiv.org">
<admin:errorReportsTo rdf:resource="mailto:biorxiv@cshlpress.edu"/>
<title>bioRxiv Subject Collection: Pharmacology And Toxicology</title>
<link>https://biorxiv.org</link>
<description>
This feed contains articles for bioRxiv Subject Collection "Pharmacology And Toxicology"
</description>

<items>
<rdf:Seq>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.15.725542v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.14.725258v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.14.725303v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.13.724950v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.12.724693v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.12.723437v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.12.724523v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.13.724891v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.13.724797v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.13.724847v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.13.724751v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.12.724311v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.12.724520v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.11.724391v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.11.724263v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.11.724251v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.11.722965v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.10.724162v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.11.724204v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.10.724063v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.12.722859v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.08.723429v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.08.723673v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.08.723702v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.07.723632v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.07.723340v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.07.723528v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.06.722533v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.07.723476v1?rss=1"/>
<rdf:li rdf:resource="https://www.biorxiv.org/content/10.64898/2026.05.07.723338v1?rss=1"/>
</rdf:Seq>
</items>
<prism:eIssn/>
<prism:publicationName>bioRxiv</prism:publicationName>
<prism:issn/>

<image rdf:resource=""/>
</channel>
<image rdf:about="">
<title>bioRxiv</title>
<url>https://www.biorxiv.org/sites/default/files/bioRxiv_article.jpg</url>
<link>https://www.biorxiv.org</link>
</image>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.15.725542v1?rss=1">
<title>
<![CDATA[
Orally Bioavailable SARS-CoV-2 Protease Inhibitors Bearing a Hydroxymethyl Ketone Warhead 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.15.725542v1?rss=1
</link>
<description><![CDATA[
The use of covalent warheads targeting the catalytic cysteine has been a cornerstone in coronavirus main protease (Mpro) inhibitor development, where various electrophilic motifs have been used including aldehydes, nitriles, ketoamides, and hydroxymethyl ketones (HMKs). Recent efforts have been mostly centered around nitrile warheads, given the success of compounds like Nirmatrelvir and Ensitrelvir in the clinic. However, finding and advancing alternative chemotypes with differentiating chemical and pharmacological profiles is essential for future pandemic preparedness. Among such alternatives, HMKs hold special interest because they balance reduced intrinsic electrophilicity with an excellent selectivity profile. Nevertheless, early HMK-based compounds, such as the clinical stage Mpro inhibitor PF-00835231, suffered from poor oral bioavailability and therefore required intravenous administration, with or without prodrug derivatization of the hydroxyl group. Here, we describe our efforts in advancing the HMK field via the discovery of mCMX110, a lead that has superior potency, increased unbound exposure in vivo, and favorable oral bioavailability in preclinical studies.
]]></description>
<dc:creator><![CDATA[ Elshan, N. G. R. D., Wolff, K. C., Weiss, F., Ghorai, S., Grabovyi, G., Wilson, K., Riva, L., Woods, A. K., Pedroarena, J., Nazarian, A., Liu, Y., Mazumdar, W., Song, L., Okwor, N., Malvin, J., Bakowski, M. A., Kirkpatrick, M. G., Gebara-Lamb, A., Huang, E., Nguyen-Tran, V. T. B., Chi, V., Li, S., Lee, K.-J., McNamara, C. W., Gupta, A. K., Rahimi, A., Chen, J. J., Joseph, S. B., Schultz, P. G., Chatterjee, A. K. ]]></dc:creator>
<dc:date>2026-05-18</dc:date>
<dc:identifier>doi:10.64898/2026.05.15.725542</dc:identifier>
<dc:title><![CDATA[Orally Bioavailable SARS-CoV-2 Protease Inhibitors Bearing a Hydroxymethyl Ketone Warhead]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.14.725258v1?rss=1">
<title>
<![CDATA[
Addiction-Like Severity Predicts Prolonged Oxycodone Withdrawal-Induced Allodynia in Genetically Diverse Rats 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.14.725258v1?rss=1
</link>
<description><![CDATA[
Opioid withdrawal is associated with heightened pain sensitivity, including allodynia. Although opioid-induced allodynia is well-documented in humans and animal models, the relationship between the severity of opioid withdrawal-induced allodynia and individual addiction-like behaviors remains poorly understood. To address this gap, Heterogeneous Stock rats underwent long access (12 hours/day) intravenous oxycodone self-administration, followed by measurement of mechanical sensitivity at six timepoints across three weeks of abstinence. Rats were stratified by an Addiction Index derived from individual differences in the escalation of oxycodone intake, motivation to consume oxycodone, tolerance to oxycodones analgesic effects, and acute withdrawal-induced mechanical pain sensitivity. Here, we show that oxycodone withdrawal induces significant and prolonged allodynia for up to three weeks, with High Addiction Index rats exhibiting greater intensity and longer duration of pain sensitivity than Low Addiction Index rats. Results remained consistent even when excluding allodynia from the Addiction Index, highlighting the robustness of the association between addiction-like severity and protracted allodynia. Linear regression associations revealed that self-administration behaviors, particularly oxycodone intake escalation and motivation to seek oxycodone, predicted subsequent withdrawal-induced allodynia severity. These findings demonstrate that greater addiction-like severity is associated with more intense and prolonged withdrawal-induced pain, supporting mechanical allodynia as a marker of addiction severity. These results motivate future work to define the mechanisms linking addiction severity to protracted opioid withdrawal-induced pain, with the goal of informing targeted clinical interventions for individuals most susceptible to severe abstinence-related allodynia.
]]></description>
<dc:creator><![CDATA[ Plasil, S. L., Tieu, L., Qian, C., Taylor, N., Sneddon, E., Carrette, L. L., Brennan, M., Morgan, A., Othman, D., Bai, K., Foroutani, S., de Guglielmo, G., Kallupi, M., George, O. ]]></dc:creator>
<dc:date>2026-05-18</dc:date>
<dc:identifier>doi:10.64898/2026.05.14.725258</dc:identifier>
<dc:title><![CDATA[Addiction-Like Severity Predicts Prolonged Oxycodone Withdrawal-Induced Allodynia in Genetically Diverse Rats]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.14.725303v1?rss=1">
<title>
<![CDATA[
Discovery of a Novel Non-MET-Mediated Otoprotective Compound Against Aminoglycoside-Induced Ototoxicity 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.14.725303v1?rss=1
</link>
<description><![CDATA[
Aminoglycoside (AG) antibiotics are indispensable for treating severe infections but frequently cause irreversible hearing loss, with no approved preventive therapies. Using in vivo zebrafish lateral line screening combined with computational scaffold-hopping, we identified a novel class of otoprotective compounds. Starting from the ion channel modulator MR16728, we discovered compound 28510 as a potent lead compound. Compound 28510 provided robust, dose-dependent protection against AG-induced hair cell damage, restoring neuromast hair cell integrity to near-control levels in acute assays and demonstrating broad efficacy across clinically relevant AGs (gentamicin, tobramycin, amikacin, streptomycin) in chronic exposures. Importantly, 28510 exhibited a favorable therapeutic window, with low micromolar 50% hair cell protection concentration (HC50) values consistently below toxicity thresholds. Mechanistically, FM1-43 and Texas Red-conjugated gentamicin uptake assays revealed that 28510 does not inhibit mechanotransduction (MET) channel-mediated AG entry, distinguishing it from current clinical candidates and pointing to a novel intracellular protective mechanism. 28510 preserved AG antibacterial activity in E. coli assays, supporting its translational compatibility as a co-therapeutic agent. Combinations of 28510 with related analogs did not yield synergistic protection; 28510 alone remained the most effective compound. In silico absorption, distribution, metabolism, and excretion (ADME) predictions further confirmed its highly favorable drug-like properties, including excellent intestinal and oral absorption. Together, these findings establish 28510 as a first-in-class, non-MET-mediated otoprotective lead with broad efficacy and a favorable therapeutic profile, highlighting a new strategy for preventing AG-induced hearing loss.
]]></description>
<dc:creator><![CDATA[ Kara, E., Nicolet, C., Rahman, S. E., Hudok, T., Leach, C., Falkner, K., Cornell, K. A., Xu, D. ]]></dc:creator>
<dc:date>2026-05-18</dc:date>
<dc:identifier>doi:10.64898/2026.05.14.725303</dc:identifier>
<dc:title><![CDATA[Discovery of a Novel Non-MET-Mediated Otoprotective Compound Against Aminoglycoside-Induced Ototoxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.13.724950v1?rss=1">
<title>
<![CDATA[
Validation and testing of an in vitro model to study medical treatments for anterior urethral stricture disease: assessing the potential efficacy of phosphodiesterase-4 (PDE4) inhibition and testosterone 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.13.724950v1?rss=1
</link>
<description><![CDATA[
Objective To 1) determine the expression and distribution of all PDE4 isozymes (A-D) along the length of the anterior urethra, 2) culture fibroblasts and epithelial cells from healthy and strictured urethras, 3) investigate an in vitro model of anterior urethral stricture disease (aUSD), and 4) assess the therapeutic potential of phosphodiesterase-4 (PDE4) inhibitors and testosterone compared to paclitaxel. Methods The presence and relative abundance of PDE4 isozymes (A-D) was confirmed using immunohistochemistry on 5 male cadaveric urethras. Human urethral fibroblasts (FBs) were cultured from healthy control urethras of patients undergoing vaginoplasty (n=3) and from idiopathic bulbar urethral strictures (L2S1E2) of patients undergoing urethroplasty (n=3). Epithelial cells (ECs) were cultured from a healthy control urethra and two urethral strictures. To investigate a model of aUSD, Control FBs were stimulated with TGFB1 and compared to Stricture FBs on assays of cell proliferation and expression of genes relevant to aUSD pathophysiology. To test therapeutics, Stricture FBs were treated with the PDE4 inhibitor, roflumilast, testosterone (T), or paclitaxel and compared to Control FBs on the previously mentioned assays and cell viability. Results PDE4- A, B, and D were detected along the length of the urethra. Expression levels did not differ between urethral regions. TGFB1 altered proliferation and gene expression in a dose-dependent manner. Roflumilast and T preserved cell viability and proliferation and decreased expression of genes positively associated with auSD. Conclusion Urethral FBs and ECs can be cultured from healthy and strictured surgical specimens, enabling in vitro research. PDE4 inhibitors and T may be non-cytotoxic alternatives or additions to paclitaxel for aUSD.
]]></description>
<dc:creator><![CDATA[ Lozano, L. P., Volk, M. J., Miller, C. D., Berg, J. E., Allamargot, C., Schlaepfer, C. H., Kurtzman, J. T., Christensen, M. B., Myers, J. B., Hertz, A. M., Swanton, A. R., Tucker, B. A., Erickson, B. A. ]]></dc:creator>
<dc:date>2026-05-17</dc:date>
<dc:identifier>doi:10.64898/2026.05.13.724950</dc:identifier>
<dc:title><![CDATA[Validation and testing of an in vitro model to study medical treatments for anterior urethral stricture disease: assessing the potential efficacy of phosphodiesterase-4 (PDE4) inhibition and testosterone]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.12.724693v1?rss=1">
<title>
<![CDATA[
Targeting Microbial Bile Salt Hydrolase Reprograms Bile Acid Metabolism and Ameliorates Metabolic Dysfunction-Associated Steatohepatitis in Mice 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.12.724693v1?rss=1
</link>
<description><![CDATA[
Microbial bile salt hydrolase (BSH) plays a central role in shaping bile acid composition and gut-liver metabolic signaling, yet its therapeutic potential in metabolic dysfunction-associated steatohepatitis (MASH) remains incompletely defined. Here, we evaluated the efficacy of the non-absorbable BSH inhibitor GR-7 in a diet induced mouse model of steatohepatitis using early and late intervention strategies with different dosing regimens. GR-7 reduced food intake and exerted stage- and dose-dependent therapeutic effects, with early intervention robustly suppressing hepatic fibrosis even at low dose, whereas late-stage administration of high-dose GR-7 markedly reduced hepatic steatosis and inflammation, as evidenced by decreased liver weight, hepatic triglyceride and cholesterol levels, and plasma ALT. Although late intervention did not result in statistically significant histological reversal of fibrosis, a trend toward improvement was observed, together with suppression of fibrogenic gene expression, suggesting that prolonged treatment may further enhance antifibrotic efficacy. Mechanistically, GR-7 effectively inhibited microbial BSH activity in vivo, leading to reduced cecal unconjugated primary and secondary bile acids-including deoxycholic acid and lithocholic acid, which was associated with improved gut barrier integrity and reduced hepatic inflammation. In parallel, BSH inhibition reprogrammed hepatic bile acid metabolism toward activation of the alternative CYP27A1-mediated synthesis pathway, accompanied by reduced food intake, thereby contributing to improved hepatic lipid accumulation. Furthermore, late-stage high-dose treatment selectively remodeled the hepatic immune landscape rather than fully restoring homeostasis, highlighting immune recalibration as a key component of therapeutic response. Together, these findings identify microbial BSH inhibition as a promising microbiome-targeted therapeutic strategy for MASH.
]]></description>
<dc:creator><![CDATA[ Wei, W., Graf, R., Wang, Y., Oalmann, C. J., Lau, J. T., Wang, X., Chien, M., Conrad, M. C., Simon, J., Ganguly, S., Yamazaki, T., Harberts, A., Chen, S., Fondevila, M. F., Dhar, D., Campbell, S. A., Senter, R. K., Schnabl, B. ]]></dc:creator>
<dc:date>2026-05-17</dc:date>
<dc:identifier>doi:10.64898/2026.05.12.724693</dc:identifier>
<dc:title><![CDATA[Targeting Microbial Bile Salt Hydrolase Reprograms Bile Acid Metabolism and Ameliorates Metabolic Dysfunction-Associated Steatohepatitis in Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.12.723437v1?rss=1">
<title>
<![CDATA[
Report on pre-validation of an animal-free alternative method (NAM) for regulatory safety testing: InfiniteLungDT, an in-vitro-learned digital twin for the prediction of material-triggered chronic neutrophilic lung inflammation 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.12.723437v1?rss=1
</link>
<description><![CDATA[
Until now, there has been no animal-free alternative method for predicting chronic inflammation and delivering the associated dose responses, the timing of onset, and the duration of inflammation, as required by regulatory agencies. Here, we demonstrate that the in-vitro-learned digital twin of neutrophilic lung inflammation (InFiniteLungDT) can obtain regulatory-relevant information about chronic neutrophilic lung inflammation by measuring dynamics of early biological effects in vitro induced by respirable materials without knowing their intrinsic properties. We constructed the digital twin(s) for each of the material, for which we have in vivo exposure data, primarily in vivo instillation data set comprising 49 different nanomaterials and 7 in vivo inhalation data set (compliant with OECD TG 412) encompassing total about 3094 single mouse exposures and 364 rat exposures (and approx. 775/225 non-exposed mouse/rat controls), to predict concentration-dependent time evolved neutrophil influx into the lung, and assess the reproducibility as well as the accuracy (predictive capacity) of both endpoints and LOAELs, that were determined at 93% for instillation, and 84% for inhalation exposure. Taking into account the time-to-deliver-result being less than 1 week, this proves that the effect of inhaled material from acute to chronic conditions can be assessed orders of magnitude faster and cheaper than in a reference animal study.
]]></description>
<dc:creator><![CDATA[ Urbancic, I., Koklic, T., Kokot, H., Kokot, B., Kozoderec, N., Kolodziej, T., Licina, T., Ma-Hock, L., Hogh Danielsen, P., Alstrup Jensen, K., Cubej Gasparin, M., Pahor, T., Cosnier, F., Valentino, S., Seidel, C., Isaxon, C., Vuk, T., Gate, L., Landsiedel, R., Stöger, T., Vogel, U. B., Strancar, J. ]]></dc:creator>
<dc:date>2026-05-17</dc:date>
<dc:identifier>doi:10.64898/2026.05.12.723437</dc:identifier>
<dc:title><![CDATA[Report on pre-validation of an animal-free alternative method (NAM) for regulatory safety testing: InfiniteLungDT, an in-vitro-learned digital twin for the prediction of material-triggered chronic neutrophilic lung inflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.12.724523v1?rss=1">
<title>
<![CDATA[
Pharmaceutical assessment of low global warming potential alternatives to HFA-134a in a budesonide, glycopyrrolate, and formoterol fumarate pressurized metered dose inhaler 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.12.724523v1?rss=1
</link>
<description><![CDATA[
Manufacturers are adopting propellants for use in pressurized metered-dose inhalers (pMDIs) that have lower global warming potentials (GWPs) than the propellants traditionally used in pMDIs. Hydrofluoroalkane (HFA)-134a has been used as the propellant in the pMDI used to deliver the fixed-dose triple combination of budesonide, glycopyrrolate and formoterol fumarate (BGF); following successful clinical evaluation, the BGF pMDI is now being transitioned to the next generation propellant hydrofluoroolefin (HFO)-1234ze(E), which has near-zero GWP. We describe formulation development efforts that led to selection of HFO-1234ze(E) over another propellant, HFA-152a, for reformulation. Propellant-specific studies evaluated active pharmaceutical ingredient (API) stability and aerodynamic particle size distribution (aPSD). Those analyses have been complemented by in silico regional lung deposition modeling conducted after the clinical evaluation of the reformulated BGF pMDI. HFO-1234ze(E) supported favorable stability and aPSD characteristics for BGF pMDI reformulation, compared with HFA-152a, and modeling predicted regional deposition consistent with therapeutic intent. Given that each pMDI is a unique combination of APIs, device, propellant, and excipients, propellant substitution requires product-specific evidence and regulatory approval, and typically takes several years. Targeted analyses, such as those described here, helped to identify the most suitable candidate propellant for successful substitution in the BGF pMDI.
]]></description>
<dc:creator><![CDATA[ Lachacz, K., Kaye, R., Mello, L., Stoker, A., Törnell, J. ]]></dc:creator>
<dc:date>2026-05-16</dc:date>
<dc:identifier>doi:10.64898/2026.05.12.724523</dc:identifier>
<dc:title><![CDATA[Pharmaceutical assessment of low global warming potential alternatives to HFA-134a in a budesonide, glycopyrrolate, and formoterol fumarate pressurized metered dose inhaler]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.13.724891v1?rss=1">
<title>
<![CDATA[
An N, S-acetylated L-cysteine-cysteamine conjugate hinders pyocyanin redox cycling to weaken Pseudomonas aeruginosa biofilm and dampens LPS-driven acute pulmonary inflammation 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.13.724891v1?rss=1
</link>
<description><![CDATA[
The persistence of P. aeruginosa infections is largely driven by the secretion of several factors during invasion, including the redox-active phenazine pyocyanin (PYO), which promotes biofilm formation and oxidative stress. Biofilms contribute to chronic infections and antibiotic resistance, limiting the efficacy of conventional therapies. We found that a synthetic compound, I-152, a conjugate of N-acetyl-L-cysteine (NAC) and S-acetylcysteamine (also known as S-acetyl-{beta}-mercaptoethylamine; SMEA), effectively restored colistin susceptibility against P. aeruginosa by altering biofilm nanomechanical properties. These perturbations in matrix integrity were associated with I-152's ability to hinder the phenazine redox cycle, shifting PYO to a reduced state and promoting chemical interactions (S-conjugates). The compound decreased PYO accumulation in bacterial cultures and PYO-generated reactive oxygen species (ROS) in macrophage cells. Together with PYO, LPS is another driver of ROS-dependent inflammatory signaling in the host, which leads to an uncontrolled cytokine response and organ damage, especially in patients with cystic fibrosis. I-152 treatment downregulated the expression of LPS-induced inflammatory cytokines, i.e., IL-6 and TNF-, in bone marrow-derived macrophages (BMDM) isolated from transgenic CFTR-/- and CFTR+/+ mice. Consistently, I-152 partially counteracted the inflammatory response in the P. aeruginosa LPS-induced acute lung injury murine model. Taken together, these results support I-152 as an adjunctive treatment for P. aeruginosa respiratory infections through a dual mechanism: combating antimicrobial resistance in biofilms and dampening host inflammation in the respiratory system.
]]></description>
<dc:creator><![CDATA[ Bruschi, M., Masini, S., Palma, F., Xiaoqiu, Y., Braga, C. L., Gregori, M., Bucci, C., Bartoccini, F., Menotta, M., Manuali, E., Minelli, L., Ligi, D., Mannello, F., Monittola, F., Zara, C., Di Pietro, C., Crinelli, R., Brandi, G., Piersanti, G., Bruscia, E. M., Schiavano, G. F., Fraternale, A. ]]></dc:creator>
<dc:date>2026-05-15</dc:date>
<dc:identifier>doi:10.64898/2026.05.13.724891</dc:identifier>
<dc:title><![CDATA[An N, S-acetylated L-cysteine-cysteamine conjugate hinders pyocyanin redox cycling to weaken Pseudomonas aeruginosa biofilm and dampens LPS-driven acute pulmonary inflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.13.724797v1?rss=1">
<title>
<![CDATA[
Sulfo-DIBMA encapsulation uniquely preserves signalling-competent active states of the class B1 GPCRs, calcitonin gene-related peptide and parathyroid hormone 1 receptors, in native-like nanodiscs 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.13.724797v1?rss=1
</link>
<description><![CDATA[
Class B1 G-protein-coupled receptors (GPCRs), such as the calcitonin gene-related peptide (CGRP) receptor and parathyroid hormone 1 (PTH1) receptor, require native lipid interactions to maintain signalling-competent conformations. However, conventional detergents disrupt these environments. Amphipathic copolymers offer a detergent-free alternative, yet the field still lacks a clear understanding of which polymer architectures best preserve active-state GPCR pharmacology, limiting their broader translational utility. Here, we examine how distinct copolymer chemistries influence the functional integrity of class B1 GPCRs by comparing SMA2000, DIBMA-12, and the electroneutral sulfo-DIBMA. Using NanoLuciferase bioluminescence resonance energy transfer (NanoBRET) ligand-binding, competition, and mini-G-protein recruitment assays on nanodisc-encapsulated receptors, we show that all three copolymers maintain high-affinity extracellular ligand binding but differ markedly in their ability to preserve intracellular signalling. Despite lower receptor extraction efficiency, only sulfo-DIBMA support mini-Gs engagement at the CGRP receptor and enable G-protein-dependent allosteric modulation at the PTH1 receptor, including conserved ligand affinity and prolonged residence time. These data reveal that polymer charge and backbone chemistry, rather than extraction yield, determine whether native-like nanodiscs retain the conformational landscape required for active-state signalling. Controlling non-specific ligand binding to the copolymer is a key requirement for a successful assay. Our findings identify sulfo-DIBMALP as a particularly superior environment for preserving native signalling behaviour in class B1 GPCRs, highlighting copolymer chemistry as an important determinant in detergent-free membrane protein studies.
]]></description>
<dc:creator><![CDATA[ Khwaja, F. N., Gunner, J., Thacker, E., Abdolhay, Y., Logan, R., Kitchen, P., Veprintsev, D., Wheatley, M., Poyner, D., Ayub, H. ]]></dc:creator>
<dc:date>2026-05-15</dc:date>
<dc:identifier>doi:10.64898/2026.05.13.724797</dc:identifier>
<dc:title><![CDATA[Sulfo-DIBMA encapsulation uniquely preserves signalling-competent active states of the class B1 GPCRs, calcitonin gene-related peptide and parathyroid hormone 1 receptors, in native-like nanodiscs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.13.724847v1?rss=1">
<title>
<![CDATA[
Effects of NMDA antagonists on social behaviour: a systematic review and meta-analysis of preclinical studies 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.13.724847v1?rss=1
</link>
<description><![CDATA[
Social withdrawal is a key component of the negative symptom domain of schizophrenia, and pharmacological blockade of the N-methyl-D-aspartate receptor (NMDAR) is widely used to model schizophrenia-relevant phenotypes in animals. However, findings on social behaviour are inconsistent across paradigms and laboratories. We therefore conducted a systematic review and meta-analysis to synthesise the effects of dizocilpine, ketamine, and phencyclidine on social interaction and social preference, to evaluate whether clinically approved antipsychotics modify these outcomes, and to examine locomotor activity measured within the same social tests to aid interpretation. We searched Embase, PubMed and Web of Science without language or date restrictions. Controlled in vivo studies in laboratory animals administering an eligible NMDAR antagonist and reporting social interaction and/or social preference outcomes were included. Two reviewers independently screened records, extracted data and assessed risk of bias. Effect sizes were computed as standardised mean differences and synthesised using correlated multilevel random-effects models with cluster-robust variance estimation. In total, 264 studies met the inclusion criteria. Overall, NMDAR antagonists were associated with reduced social interaction and reduced social preference relative to controls, although the social preference literature appeared vulnerable to small-study effects and imprecision. Locomotor activity measured during social interaction tests tended to be higher following NMDAR antagonists, whereas during social preference no consistent overall change was observed. In animals exposed to NMDAR antagonists, antipsychotics increased social behaviour, but these changes commonly co-occurred with reduced locomotion during social interaction tests, suggesting that improvements in social measures may partly reflect altered behavioural competition and time allocation rather than selective restoration of social functioning. Taken together, the evidence supports an overall link between NMDAR antagonism and reduced social behaviour, but the strength and interpretability of this signal depend on the paradigm and are constrained by heterogeneity and limitations in reporting.
]]></description>
<dc:creator><![CDATA[ Gallas-Lopes, M., Abreu, M. B., Andrades, M., Arbo, B. D., Bastos, L. M., Caetano, T. C., Muller, D. V., Patelli-Alves, A., Rosa, D. A., Stein, D. J., Herrmann, A. P. ]]></dc:creator>
<dc:date>2026-05-15</dc:date>
<dc:identifier>doi:10.64898/2026.05.13.724847</dc:identifier>
<dc:title><![CDATA[Effects of NMDA antagonists on social behaviour: a systematic review and meta-analysis of preclinical studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.13.724751v1?rss=1">
<title>
<![CDATA[
Topological Pharmacokinetics: Reading the Shape of Drug Disposition from Data 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.13.724751v1?rss=1
</link>
<description><![CDATA[
Pharmacokinetic analysis has spent half a century compressing drug concentration time curves into scalar summaries (AUC, Cmax, clearance) discarding the shape information that encodes mechanistic fingerprints of the underlying physiology. We introduce Topological Pharmacokinetics (TPK), a framework that reads the shape of pharmacokinetic trajectories directly from data without prior commitment to a compartmental model. TPK uses delay embedding to reconstruct the pharmacokinetic attractor from the concentration time curve, and persistent homology to extract its topological invariants connected components and loops as a Pharmacokinetic Topological Invariant (PTI) vector. We validate TPK across three levels: linear systems (negative control), nonlinear saturable elimination (detection of the N(PTP)+1 rule and a nonlinear diagnostic triad), and endogenous circadian rhythms (contrastive detection of rhythmic interference via Dev specificity and Decouple Collapse). The PTI vector provides a model agnostic shape fingerprint that, in simulation, demonstrates the diagnostic potential of shape based analysis; validation on experimental data is required to assess whether this potential generalizes to real pharmacokinetic data. All findings are demonstrated as proof of concept on simulated data; validation on experimentally measured concentration time curves is the essential next step.
]]></description>
<dc:creator><![CDATA[ Ren, H.-C., Gu, Y.-X. ]]></dc:creator>
<dc:date>2026-05-15</dc:date>
<dc:identifier>doi:10.64898/2026.05.13.724751</dc:identifier>
<dc:title><![CDATA[Topological Pharmacokinetics: Reading the Shape of Drug Disposition from Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.12.724311v1?rss=1">
<title>
<![CDATA[
A novel pipeline for the validation of manganese chelators for the treatment of manganese overload 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.12.724311v1?rss=1
</link>
<description><![CDATA[
Manganese neurotoxicity, arising from environmental overexposure or inherited transporter disorders due to pathogenic variants in SLC30A10 and SLC39A14, leads to manganism, a debilitating Parkinsonian movement disorder. Alhtough chelation therapy can partially reverse neuropathology, current clinical practice relies on intravenous CaNa2EDTA, which is burdensome and poorly suited for long-term use. Consequently, there remains a significant unmet need for more effective, orally bioavailable chelators. This study aimed to establish and validate a pipeline for identifying and assessing novel ligands that attenuate manganese neurotoxicity and support preclinical translational development.

Based on the structural features of manganese-based MRI contrast agents, we selected two chelators, N-picolyl-N,N',N'-trans-1,2-cyclohexylenediaminetriacetic acid (H3PyC3A) and ethylenediaminetetraacetic acid-benzothiazole aniline (H4EDTA-BTA), and their methyl ester derivatives, Me3PyC3A and Me4EDTA-BTA. These were evaluated in vivo using zebrafish (slc39a14U801/U801) and mouse (Slc30a10KO/KO) models of manganese overload.

H3PyC3A and Me3PyC3A demonstrated greater manganese-mobilizing efficacy than CaNa2EDTA, improving locomotor behavior in slc39a14U801/U801 zebrafish. In Slc30a10KO/KO mice, intravenous administration confirmed selective in vivo chelation of excess manganese over physiological concentrations of zinc and copper. Although oral bioavailability was low (<1%), long-term oral administration of H3PyC3A modestly reduced liver and brain Mn accumulation, suggesting an added benefit of oral administration via gastrointestinal chelation.

This integrated in vitro to in vivo pipeline provides a robust and scaleable approach for the development of next-generation Mn chelators. Slc39a14U801 loss-of-function zebrafish enable high throughput identification of candidate compounds while Slc30a10KO/KO mice offer a clinically relevant disease model for pharmacokinetic profiling and proof-of-concept validation.
]]></description>
<dc:creator><![CDATA[ Vogt, H., Pojani, C., Devonport, J., McGown, A., Firth, G., Doykov, I., Nikolaenko, V., Anagianni, S., Valdivia, L. E., Khalil, Y., Bodnar, N., Kallay, C., Dadswell, C., Gonzalez-Mendez, R., Purchase, R., Platt, F. M., Zacconi, F. C. M., Geard, A. F., Heywood, W. E., Mills, K., Mills, P. B., Rahim, A. A., Rihel, J., Wilson, S. W., Kostakis, G. E., Spencer, J., Tuschl, K. ]]></dc:creator>
<dc:date>2026-05-15</dc:date>
<dc:identifier>doi:10.64898/2026.05.12.724311</dc:identifier>
<dc:title><![CDATA[A novel pipeline for the validation of manganese chelators for the treatment of manganese overload]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.12.724520v1?rss=1">
<title>
<![CDATA[
A Cherry-Flavoured E-Cigarette Adduct, BPGA, Reprograms Alveolar Epithelial Cell Fate Through Epithelial-to-Mesenchymal Transition and Evasion of Apoptosis 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.12.724520v1?rss=1
</link>
<description><![CDATA[
E-cigarettes have attracted significant attention as a safer substitute for conventional tobacco smoking. However, they have introduced new inhalable toxicants, including benzaldehyde-propylene glycol acetal (BPGA)--a chemical adduct produced by cherry-flavoured e-cigarettes. The health risks associated with such flavour-derived acetals remain insufficiently elucidated at the cellular level. This study investigated the role of BPGA in the progression of epithelial-to-mesenchymal transition (EMT)-like changes in alveolar epithelial cells (A549 cells).

A549 cells exposed to various concentrations of BPGA were analysed for cell viability, morphology, mitochondrial function, lysosomal health, and cytoskeletal integrity using viability assays and fluorescence imaging. Intracellular reactive oxygen species (ROS) production was quantified using the 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) assay. Antioxidant enzyme expression, inflammatory responses, and EMT-associated phenotypic alterations were evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunofluorescence (IF) assays.

Exposure of alveolar epithelial cells to BPGA caused a concentration-dependent decrease in cell viability. BPGA exposure resulted in mitochondrial membrane depolarisation, lysosomal damage, cytoskeletal changes, and stress fibre formation, which altered cell morphology. It significantly increased intracellular ROS production. As a result, antioxidant enzyme levels were upregulated as a protective response. However, during severe oxidative stress, this response was overwhelmed. Excess ROS disrupted cellular homeostasis and initiated apoptosis, though not completely. ROS also acted as a signalling molecule, promoting the upregulation of inflammatory mediators. These changes were associated with altered EMT marker expression, suggesting that BPGA might drive EMT-like remodelling.

In conclusion, BPGA, a chemical adduct from e-cigarette vapour, induces alveolar injury by promoting oxidative stress, inflammation, and EMT-related changes, which may explain a mechanism by which e-cigarette exposure could lead to lung injury and pulmonary fibrosis.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=169 SRC="FIGDIR/small/724520v1_ufig1.gif" ALT="Figure 1">
View larger version (60K):
org.highwire.dtl.DTLVardef@d6b1c5org.highwire.dtl.DTLVardef@1312700org.highwire.dtl.DTLVardef@1f777c4org.highwire.dtl.DTLVardef@1d7f020_HPS_FORMAT_FIGEXP  M_FIG O_FLOATNOGraphical abstractC_FLOATNO C_FIG
]]></description>
<dc:creator><![CDATA[ Xavier, J., Yu, Y., Varma, B., Lu, Z., KB, M., NS, R., PR, A. K., Bernardino de la Serna, J. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.12.724520</dc:identifier>
<dc:title><![CDATA[A Cherry-Flavoured E-Cigarette Adduct, BPGA, Reprograms Alveolar Epithelial Cell Fate Through Epithelial-to-Mesenchymal Transition and Evasion of Apoptosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.11.724391v1?rss=1">
<title>
<![CDATA[
ERRγ deletion in podocytes accelerates aging related kidney disease 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.11.724391v1?rss=1
</link>
<description><![CDATA[
We have recently demonstrated that treatment of aged mice with a pan-ERR agonist reverses age-related increase in urinary albumin, decrease in podocyte density, impaired mitochondrial function, and inflammation. The contribution of individual isoforms of ERRs however has not been determined. Since the aging kidney showed a possible compensatory increased expression of ERR{gamma} in the podocytes, in the face of decreased ERR expression, in the present study we aimed to determine the role of ERR{gamma} in aging podocyte. To this end, we cross bred ERR{gamma} floxed mice with podocin-Cre mice to achieve a podocyte-specific ERR{gamma} deletion. While these mice at 3 months of age showed no effect on albuminuria compared to the wild type, when the mice were aged to 21 months of age, there was a significant increase in albuminuria and decrease in podocyte density. Furthermore, we found that the podocyte deletion of ERR{gamma} primarily targeted the expression of mitochondrial biogenesis regulator PGC-1, and mitochondrial fatty acid oxidation enzymes CPT1a and MCAD in the kidney. Electron Microscopy (EM) revealed thickened glomerular basement membrane and diffuse podocyte foot process effacement, as well as severe mitochondrial damage including cristae abnormalities, fragmentation, and changes indicative of altered fusion and fission dynamics. Fluorescence Lifetime Imaging Microscopy (FLIM) to determine NADH and FAD lifetimes indicate a metabolic shift from mitochondrial oxidative phosphorylation towards glycolysis, and decrease in mitochondrial redox capacity. Considering a significantly decreased expression of ERR in aging podocytes plus its traditional role in mitochondrial function, these studies using podocyte ERR{gamma} deletion suggested an overlapping mechanism for ERR/ERR{gamma} to act as modulators of age-related mitochondrial dysfunction and age-related kidney disease.
]]></description>
<dc:creator><![CDATA[ Wang, X. x., Myakala, K., Shults, N. V., Penjweini, R., Clarkson-Paredes, C., Krawczyk, E., Hegde, S., Popratiloff, A., Panov, J., Fan, R., Guthrie, G., Yang, X. P., Rosenberg, A. Z., Knutson, J., Levi, M. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.11.724391</dc:identifier>
<dc:title><![CDATA[ERRγ deletion in podocytes accelerates aging related kidney disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.11.724263v1?rss=1">
<title>
<![CDATA[
Human internal exposures to alternariol and its monomethyl ether are predicted below thresholds of in vitro toxicity by physiologically based kinetic modeling 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.11.724263v1?rss=1
</link>
<description><![CDATA[
The foodborne mycotoxins alternariol (AOH) and alternariol monomethyl ether (AME) have been associated with several adverse effects, including cytotoxicity, genotoxicity, endocrine disruption, and immunomodulation. As these endpoints are typically observed in vitro at micromolar concentrations, the question arises whether such levels are attainable in exposed humans. To address this data gap in chemical risk assessment, a physiologically based kinetic (PBK) model was developed to predict internal exposure doses to AOH and AME in humans.

As input parameters, kinetic constants for hepatic glucuronidation were obtained in vitro by incubating Sprague Dawley rat and human liver S9 fractions with 0.5-50 M AOH and 0.5-20 M AME, demonstrating rapid biotransformation in both species. Intestinal absorption of AME and physicochemical parameters were estimated using quantitative structure-activity relationship (QSAR) models. Sensitivity analysis identified parameters describing hepatic glucuronidation and gastrointestinal uptake as among the most influential, confirming the importance of their reliable estimation. The PBK model was evaluated against available rodent toxicokinetic data and subsequently extrapolated to humans. Ultimately, the currently available exposure estimates published by EFSA in 2016 were applied to predict target tissue concentrations, which were compared to points of departure (PoDs) for relevant toxicological endpoints.

Even in the most susceptible group of male toddlers, predicted internal concentrations (10-4 M range) were approximately four orders of magnitude below the respective PoDs. Consequently, under the applied exposure assumptions and considering the compounds as isolated chemicals, AOH and AME are not expected to reach systemic or tissue concentrations associated with the investigated effects.
]]></description>
<dc:creator><![CDATA[ Borsos, E., Descamps, B., Hetzschold, N., Varga, E., Marko, D., Aichinger, G. ]]></dc:creator>
<dc:date>2026-05-14</dc:date>
<dc:identifier>doi:10.64898/2026.05.11.724263</dc:identifier>
<dc:title><![CDATA[Human internal exposures to alternariol and its monomethyl ether are predicted below thresholds of in vitro toxicity by physiologically based kinetic modeling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.11.724251v1?rss=1">
<title>
<![CDATA[
Comparative metabolism of the Alternaria toxins altenuene and tentoxin in rat and human primary hepatocytes 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.11.724251v1?rss=1
</link>
<description><![CDATA[
The ubiquitously occurring food contaminants altenuene (ALT) and tentoxin (TEN) are recognized as emerging Alternaria mycotoxins, yet substantial data gaps remain when it comes to their toxicological behavior and toxicokinetic characteristics. This study aimed to compare and generate quantitative data on their hepatic metabolism and to obtain semi-quantitative insights into their metabolite profiles. To this end, primary rat and human hepatocytes were incubated with 10 {micro}M ALT or TEN over multiple time points up to 4 h. Both substrate depletion and metabolite identification revealed pronounced interspecies differences. The extent of ALT metabolism was significant, with an 88% and 57% decrease in rat and human hepatocytes after 4 h, respectively. In contrast, TEN showed extensive biotransformation in rats (67%) but only modest turnover in humans (27%) over the same period. Hepatocellular clearances were consistently higher for ALT than TEN, with hepatic extraction ratios indicating intermediate extraction for ALT and low extraction for TEN. High-resolution mass spectrometry combined with targeted analysis of selected metabolites annotated phase II conjugation as the predominant metabolic pathway for ALT and phase I oxidative metabolism for TEN, including mono- and double-metabolized species for the latter. Overall, these results provide a comprehensive characterization of ALT- and TEN-metabolism in hepatocytes, offering a foundation for future studies on their toxicological relevance and impact on human health.
]]></description>
<dc:creator><![CDATA[ Borsos, E., Gendre, C., Mahdjoub, M., Varga, E., Dubreil, E., Henri, J., Le Hegarat, L., Marko, D. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.11.724251</dc:identifier>
<dc:title><![CDATA[Comparative metabolism of the Alternaria toxins altenuene and tentoxin in rat and human primary hepatocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.11.722965v1?rss=1">
<title>
<![CDATA[
Drug Proarrhythmic Evaluation in a High Throughput Cardiac New Approach Methodology 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.11.722965v1?rss=1
</link>
<description><![CDATA[
Background and PurposeCardiotoxicity is a major cause for drug failure throughout the drug development process, with particular concern for action potential prolongation and arrhythmia. Hence, such liabilities are heavily considered during the early phases of drug design to pre vent dangerous compounds from progressing. New approach methodologies (NAMs) that efficiently examine this risk early in the discovery pipeline should help streamline drug development programs. We developed a cardiac NAM, a 384-well open bath platform consisting of cardiac tissue derived from human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes, enabling high-throughput drug screening while maintaining the structural and functional complexity of 3D cardiac micromuscles.

MethodsWe dramatically increased throughput without compromising physiological relevance provided by the 3D micromuscle structure. Our 384-well open bath high-throughput platform allowed evaluation of multiple compounds at a time, enabling us to study the CiPA (comprehensive in vitro proarrhythmia assay) drug panel for proarrhythmia screening. We obtained phenotypic fingerprints of all 28 compounds (9 low, 11 intermediate, and 8 high arrhythmia risk; https://cipaproject.org) in dose-escalation studies around their respective clinical concentrations. The analysis was augmented with an in silico pipeline that used phenotypic biomarkers to invert data into a mathematical model of cellular currents to infer which ion channels were affected upon drug exposure, and then trained a ML model to predict channel block.

Results and ConclusionsWe found accurate detection of arrhythmic potential for most of the compounds, and the in silico model inversions were consistent with published values of compound channel block. All the high risk compounds showed action potential duration (APD) prolongation coupled with either action potential abnormalities, early afterdepolarizations (EADs), or beat cessation. For the intermediate risk group, 9 out of 11 compounds caused APD prolongation alone or in combination with EADs while 2 others showed either beat cessation or beat rate change. Augmentation of APD analysis with detailed biophysical modeling and ML tools provided meaningful insight into the mechanisms involved in APD changes. Overall, our cardiac NAM allowed for fast and relevant screening for mechanistic understanding of APD prolongation and proarrhythmic activity, at massively increased throughput compared to other 3D micromuscle models.

SummaryCardiotoxicity testing is critical in drug development to prevent arrhythmogenic side effects. Current stringent regulations have greatly reduced market withdrawals; however, these strict evaluations often lead to costly late-stage failures and loss of promising candidates as false positives. We developed a cardiac new approach methodology (NAM), a 384-well open bath cardiac micromuscle platform created from hiPSC-derived cardiomyocytes, enabling high-throughput drug screening while maintaining the structural and functional complexity of 3D cardiac micromuscles. Using the comprehensive in vitro proarrhythmia assay (CiPA) drug panel, we validated the system to accurately detect proarrhythmic potential. Our assay provided phenotypic fingerprints based on mechanical and electrophysiological biomarkers. Integration with computational modeling offered insights into multi-ion channel effects (MICE). Particularly, we identified sodium channel block contributions as a significant factor for poor risk prediction based on traditional parameters. The combined experimental and computational platform can enhance early drug screening, thereby reducing late-stage failures and promoting the progression of low-risk compounds with complex electrophysiological profiles.
]]></description>
<dc:creator><![CDATA[ Charwat, V., Ramirez, A., Jaeger, K. H., Kandalaft, B., Finsberg, H., Siemons, B., Tveito, A., Healy, K., Wall, S. T. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.11.722965</dc:identifier>
<dc:title><![CDATA[Drug Proarrhythmic Evaluation in a High Throughput Cardiac New Approach Methodology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.10.724162v1?rss=1">
<title>
<![CDATA[
Toward a Random Background for Ligand Optimization 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.10.724162v1?rss=1
</link>
<description><![CDATA[
Ligand optimization is central to drug discovery as hundreds of analogs might be designed and synthesized between an initial hit and a therapeutic candidate. The efficiency of this process is unclear, at least partly because there is no random background for optimization against which to compare. Such a random background might emerge from synthetically accessible but otherwise systematic random small substitutions across starting ligands, measuring likelihood of achieving a substantial improvement in affinity/potency or other property by any single perturbation. Recent literature and ligand-affinity/potency databases suggest that perhaps 10% of analogs with minor modifications improve upon a parents potency substantially (by [&ge;]10-fold), but this number is clouded by reporting bias, intentional improvement, and inter-group reproducibility. To begin to establish a background expectation for ligand optimization, we comprehensively and systematically modified 18 lead molecules across six targets with single atom changes; 257 compounds were synthesized. Unexpectedly, 11.2% of these random small perturbation analogs improved potency by [&ge;]10-fold over their parents. Conversely, these more potent analogs typically had worse in vitro pharmacokinetics (e.g. reduced metabolic stability, lower plasma free fraction). While it was possible to find analogs where the potency increase compensated for inferior exposure and half-life, resulting in more potent compounds in vivo, overall a frustrated landscape for ligand optimization is revealed. This study begins to establish a background expectation for ligand potency optimization and offers a simple strategy to do so. It also begins to quantify the challenges confronting the field in moving beyond in vitro potency.
]]></description>
<dc:creator><![CDATA[ Xu, X., Mailhot, O., Correy, G. J., Huang, X., Braz, J., Shi, D., Srinivasan, K., Zielinski, K., Holota, Y., Kuziv, Y., Tsoutsouvas, C., Levinzon, N., Doruk, Y. U., Rachman, M., Diolaiti, M., Stevens, M., Liu, F., Holland, K., Hubner, H., Wang, J., Wu, Y., Ashworth, A., Makriyannis, A., Zhang, Y., Moroz, Y., Gmeiner, P., Abel, R., Manglik, A., Basbaum, A. I., Roth, B. L., Fraser, J. S., Shoichet, B. K. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.10.724162</dc:identifier>
<dc:title><![CDATA[Toward a Random Background for Ligand Optimization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.11.724204v1?rss=1">
<title>
<![CDATA[
Adequate coating and change in morphology increase the performances of silver nanoparticular biocides 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.11.724204v1?rss=1
</link>
<description><![CDATA[
Silver has been used as a biocide for centuries, mostly in health-oriented applications. However, as a biocide, silver is toxic not only to its intended targets, mainly bacteria and fungi, but also to all living cells. Because of this toxicity, it is desirable to use forms of silver that maximize the required biocidal activity while minimizing the amount of silver that will be released in the environment at the end of life of the product. Silver nano objects are a good compromise for such requirements. The high surface to volume ratio allows for good reactivity and thus good biocidal activity, while the small amount of silver present in nano objects allows for a limited environmental release at the product end of life. In this work, we tested three types of silver nano objects. The first type, polyvinylpyrrolidone-coated silver nanoparticles (nAg-PVP) were used as a control nanoparticle, as this type of nanoparticle is now widespread. We also manufactured and tested maltodextrin-coated silver nanoparticles (nAg-MD) and micrometric (20 {micro}m in two dimensions and a few nanometers in the third one) silver flakes ({micro}AgSF). For these three silver nano objects, we investigated the biocidal activity by stringent tests using both Staphylococcus aureus and Escherichia coli as target bacteria. In addition, we investigated toxicity on mammalian macrophages or keratinocytes cell lines, as well as on an insect hemocyte cell line. Our results showed that the two innovative silver nano objects (nAg-MD and even more {micro}AgSF), showed both a better bactericidal activity and a lesser toxicity than the reference nAg-PVP nanoparticles. In addition, we also checked that beyond toxicity, the silver nano objects did not induce an inflammatory reaction, making them safer to use.
]]></description>
<dc:creator><![CDATA[ Pawłowski, B., Błazyca, H., Huotari, J., Collin, V., Chartier-Garcia, E., Salo, S., Darrouzet, E., Jeremiasz, O., Rabilloud, T. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.11.724204</dc:identifier>
<dc:title><![CDATA[Adequate coating and change in morphology increase the performances of silver nanoparticular biocides]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.10.724063v1?rss=1">
<title>
<![CDATA[
Cyclic Peptides Target CAPON and Modulate Cellular Responses under Alzheimers Disease-Relevant Stress 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.10.724063v1?rss=1
</link>
<description><![CDATA[
CAPON (NOS1AP) is an adaptor protein involved in neuronal nitric oxide synthase (nNOS) signaling and has been implicated in Alzheimers disease (AD), excitotoxicity, and tau-associated neurodegeneration. Here, we report the identification of cyclic peptide ligands targeting CAPON using phage display screening of a disulfide-constrained peptide library. Phage enrichment, ELISA validation, microscale thermophoresis (MST), and biolayer interferometry (BLI) identified CAP1 as the lead peptide, exhibiting low micromolar binding affinity toward CAPON. Computational studies further supported stable CAPON-CAP1 interactions through complementary hydrophobic and electrostatic contacts. Functionally, CAP1 attenuated A{beta}42-induced neuronal toxicity, suppressed NMDA-driven nitric oxide production, and reduced pathological tau phosphorylation in neuronal models under AD-relevant stress conditions. In addition, CAP1 demonstrated favorable preliminary pharmacokinetic properties, including good aqueous solubility, plasma stability, and measurable membrane permeability. Collectively, these findings establish the first cyclic peptide ligands targeting CAPON and identify CAP1 as a promising scaffold for modulation of CAPON-dependent neurodegenerative signaling.
]]></description>
<dc:creator><![CDATA[ Abdo, A., Yuan, S., Kuncewicz, K., Mo, J., Duan, H., Gabr, M. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.10.724063</dc:identifier>
<dc:title><![CDATA[Cyclic Peptides Target CAPON and Modulate Cellular Responses under Alzheimers Disease-Relevant Stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.12.722859v1?rss=1">
<title>
<![CDATA[
Integrated luminescence and phenotypic profiling for drug discovery in a zebrafish model of Marfan syndrome 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.12.722859v1?rss=1
</link>
<description><![CDATA[
BackgroundMarfan syndrome (MFS) is a life-threatening heritable connective tissue disorder caused by pathogenic variants in fibrillin-1, characterized by progressive cardiovascular disease. Current medical therapies slow disease progression but do not prevent major complications, underscoring the need for new treatment strategies and unbiased discovery approaches.

MethodsWe used a zebrafish model of MFS lacking fibrillin-3 (fbn3-/-), which recapitulates key cardiovascular phenotypes including cardiac stress, valvular defects, arrhythmia, and aortic dilation. To enable sensitive, quantitative assessment of cardiac stress, we generated a novel transgenic zebrafish reporter expressing secreted nanoluciferase under control of the stress-responsive nppb promoter. This reporter was combined with morphological phenotyping and bulbus arteriosus (BA) imaging. We evaluated standard MFS therapies, targeted modulators of TGF-{beta} signaling, and performed an unbiased high-throughput drug screen of over 1 500 clinically approved compounds across multiple developmental treatment windows.

Resultsfbn3-/- larvae exhibited markedly elevated nppb activity that correlated with phenotypic severity and peaked during stages of highest mortality. The nanoluciferase reporter provided a [~]1 000-fold dynamic range, substantially outperforming Firefly luciferase-based assays. Pharmacological inhibition of TGF-{beta} signaling produced transient or deleterious effects, while {beta}-blockers, losartan, and allopurinol failed to consistently improve cardiac stress, pericardial edema, or BA dilation. The unbiased high-throughput drug screen identified a small number of primary and secondary hits; however, none demonstrated reproducible phenotypic rescue upon rigorous multi-dose, multi-time window validation.

ConclusionsThis study establishes a sensitive zebrafish-based platform for early, quantitative assessment of cardiovascular stress in MFS. Our findings highlight the limited efficacy of current therapies, the context-dependent nature of TGF-{beta} modulation, and the biological complexity underlying MFS pathogenesis. Although no definitive therapeutic candidates were identified, this work lays a robust foundation for expanded unbiased discovery efforts aimed at identifying disease-modifying interventions for MFS.
]]></description>
<dc:creator><![CDATA[ Horvat, M., Caboor, L., De Rycke, K., Mennens, L., Daniels, E., Wyseur, J., Verhelst, E., Roos, I., Rodriguez-Rovira, I., Egea, G., De Backer, J., Sips, P. ]]></dc:creator>
<dc:date>2026-05-13</dc:date>
<dc:identifier>doi:10.64898/2026.05.12.722859</dc:identifier>
<dc:title><![CDATA[Integrated luminescence and phenotypic profiling for drug discovery in a zebrafish model of Marfan syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723429v1?rss=1">
<title>
<![CDATA[
Acute inflammation-mediated attenuation of behavioural sensitization in methamphetamine-sensitized mice via distinct COX-2 and TNF-α pathways 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723429v1?rss=1
</link>
<description><![CDATA[
Background and PurposeWhile inflammation has been generally considered to exacerbate symptoms of schizophrenia, some clinical observations suggest that acute inflammation may alleviate positive symptoms. However, animal models often use excessive inflammatory stimuli, and the effects of acute inflammation--comparable to levels observed in patients--remain unknown.

Experimental ApproachTo address this, we examined whether acute inflammation induced under relatively mild, clinically relevant conditions suppresses behavioural sensitization in methamphetamine (METH)-sensitized mice, a model of psychostimulant-induced psychosis with relevance to certain aspects of positive symptoms of schizophrenia. We used a repeated METH (1 mg/kg) sensitized model to evaluate the effects of acute inflammation on behavioural sensitization. Acute inflammation was induced via two methods using either lipopolysaccharides (LPS; 1 g/kg) to mimic peripheral immune activation or restraint stress (RS; single 2-h exposure) to model the neuroinflammation induced by psychological stress. LPS doses were adjusted with reference to the magnitude of peripheral cytokine elevation reported in patients, and RS was applied in short single sessions to avoid excessive inflammation.

Key ResultsBoth LPS and RS significantly suppressed behavioural sensitization, without inducing other behavioural abnormalities. This suppression was dependent on toll-like receptor-4 activation. LPS-mediated suppression involved cyclooxygenase-2, whereas RS-mediated suppression was linked to the microglia-derived tumour necrosis factor-. LPS did not alter, whereas RS significantly reduced the striatal extracellular dopamine levels.

Conclusion and ImplicationsThese findings suggest that acute inflammation suppresses behavioural sensitization through distinct mechanisms depending on the inflammatory trigger, providing a framework for understanding how inflammation may influence psychosis-related processes, with potential relevance to schizophrenia.
]]></description>
<dc:creator><![CDATA[ Shinohara, R. C., Ishikawa, S., Matsumoto, R., Ito, K., Tonosaki, M., Matsuyama, S., Ohgidani, M., Koga, M., Hashimoto, N., Kusumi, I., Takahiro, K. A. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723429</dc:identifier>
<dc:title><![CDATA[Acute inflammation-mediated attenuation of behavioural sensitization in methamphetamine-sensitized mice via distinct COX-2 and TNF-α pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723673v1?rss=1">
<title>
<![CDATA[
Ramelteon facilitates hippocampal ripple occurrence and amplitude in mice 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723673v1?rss=1
</link>
<description><![CDATA[
The hippocampus is essential for memory consolidation, a process mediated by high-frequency oscillations known as ripples during non-rapid eye movement (NREM) sleep. Ramelteon, a selective MT1/MT2 receptor agonist, has been reported to possess cognitive-enhancing properties; however, its impact on the fine-scale dynamics of hippocampal ripples remains unclear. We performed chronic local field potential recordings from the dorsal hippocampus and prefrontal cortex in mice. Following the intraperitoneal administration of either vehicle or ramelteon, we evaluated sleep architecture and characterized ripple properties, including occurrence rate, amplitude, instantaneous frequency, and duration during NREM sleep. Ramelteon administration significantly increased NREM sleep occupancy. Notably, we found that ramelteon significantly enhanced both the occurrence rate and amplitude of hippocampal ripples compared to the control group. While a slight increase in intra-ripple frequency was observed, other structural features, such as ripple duration and asymmetry index, remained unaffected. Our findings demonstrate that ramelteon facilitates hippocampal ripple dynamics by increasing their occurrence and synchrony during NREM sleep. Given the critical role of ripples in memory consolidation, these neurophysiological changes may underlie the procognitive effects of ramelteon.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=55 SRC="FIGDIR/small/723673v1_ufig1.gif" ALT="Figure 1">
View larger version (15K):
org.highwire.dtl.DTLVardef@17af46dorg.highwire.dtl.DTLVardef@145376eorg.highwire.dtl.DTLVardef@13a49acorg.highwire.dtl.DTLVardef@1bae6ea_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator><![CDATA[ Nakashima, M., Miyano, M., Kuroyanagi, H., Sasahara, A., Ikegaya, Y., Matsumoto, N. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723673</dc:identifier>
<dc:title><![CDATA[Ramelteon facilitates hippocampal ripple occurrence and amplitude in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723702v1?rss=1">
<title>
<![CDATA[
Activation by statins unveils two putative agonist binding sites in the pore domain of TRPA1 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723702v1?rss=1
</link>
<description><![CDATA[
TRPA1 is a non-selective cation channel that plays a crucial role in several pain and inflammatory conditions. Agents reducing membrane cholesterol decrease TRPA1 activation, but it remains unclear how cholesterol-lowering medications affect TRPA1 function. Given that TRPA1 is activated by a wide variety of chemicals, we explored whether statins have acute effects on this channel. We found that five commonly used statins activate human and mouse TRPA1 in a reversible and concentration-dependent manner. The effective concentrations were above the micromolar range, in the order: simvastatin {approx} lovastatin < fluvastatin < atorvastatin < pravastatin. Statin-induced activation was not correlated to changes in membrane order, nor mediated by N-terminal cysteine residues contributing to electrophilic compound agonism. Molecular docking calculations and the functional characterization of single-point mutants revealed two separate putative binding sites, one situated close to the kink of transmembrane segment 5 (TM5) and the other at the interface between TM4 and TM5. The mTRPA1 inhibitor A-967079 largely abrogated the response to the electrophilic agonist allyl isothiocyanate, but had weaker and varied effects across different statins and menthol. Mutation T877L strongly altered the effect of A-967079, also in an agonist-dependent manner, suggesting competitive binding between this antagonist and the non-electrophilic agonists. The identification of two distinct agonist binding sites may help explaining how TRPA1 is able to respond to a large variety of non-electrophilic compounds, while the finding of competitive interactions at one of these sites may help guide the development of agonist-specific antagonists of therapeutic relevance.
]]></description>
<dc:creator><![CDATA[ Startek, J. B., Milici, A., Held, K., Talavera, A., Talavera, K. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723702</dc:identifier>
<dc:title><![CDATA[Activation by statins unveils two putative agonist binding sites in the pore domain of TRPA1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723632v1?rss=1">
<title>
<![CDATA[
Virtual screening and zebrafish phenotype-based evaluation argues against repurposing 4-phenylbutyrate for STXBP1-relateddisorders 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723632v1?rss=1
</link>
<description><![CDATA[
Syntaxin-binding protein 1 (STXBP1) mutations lead to severe epilepsy, intellectual disability, developmental delay, and movement disorder. Effective treatments for these conditions do not exist. Recent studies in Munc18-1 (STXBP1) C. elegans models demonstrate that 4-phenylbutyrate (4-PBA) or related pharmacological chaperones stabilize Munc18-1 protein levels and rescue locomotion deficits. These studies suggest a novel treatment strategy for these patients. Here, we used a stxbp1a zebrafish model with a profound movement disorder to screen 4-PBA and alternative structural analogs identified using artificial intelligence (AI)-based screening. Automated locomotion assays conducted on larval stxbp1a mutant zebrafish at 5 days post-fertilization (dpf) confirm and extend the movement disorder endophenotype. Drug treatment (4-PBA or 16 identified candidates) failed to rescue the stxbp1a mutant zebrafish locomotion deficit. Electrophysiology studies in a stxbp1b zebrafish model characterized by spontaneous seizure activity (i.e., epilepsy) failed to detect a reduction in ictal-like events with 4-PBA treatment. Taken together, our results suggest caution in repurposing 4-PBA or related compounds for treatment of STXBP1 disorders.
]]></description>
<dc:creator><![CDATA[ Frick, A., Whyte-Fagundes, P. C., Baraban, S. C. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723632</dc:identifier>
<dc:title><![CDATA[Virtual screening and zebrafish phenotype-based evaluation argues against repurposing 4-phenylbutyrate for STXBP1-relateddisorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723340v1?rss=1">
<title>
<![CDATA[
Nephrotoxicity of Immune Checkpoint Inhibitors in Mice with a Human Immune System 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723340v1?rss=1
</link>
<description><![CDATA[
IntroductionImmune checkpoint inhibitors (ICIs) enhance antitumor responses by blocking inhibitory receptors, including PD-1 and CTLA-4. Overactivation can trigger systemic toxicity akin to autoimmune diseases, including kidney manifestations. We sought to 1) profile immune signaling and 2) interrogate potential mechanisms of ICI-related kidney injury in a Human Immune System (HIS) tumor-bearing mouse model treated with nivolumab and ipilimumab.

MethodsImmunodeficient BRGS (BALB/c-Rag2nullIl2r{gamma}nullSirpNOD) neonates were engrafted with human CD34+ cells to generate HIS-BRGS mice. Human MDA-MB-231 tumor cells were implanted subcutaneously; once tumors reached [~]150 mm3, mice received weekly intraperitoneal vehicle (PBS) or ICI (nivolumab 20 mg/kg + ipilimumab 10 mg/kg) for 4 weeks (Veh BRGS n=4; ICI BRGS n=6; Veh HIS-BRGS n=7; ICI HIS-BRGS n=7). Kidneys were evaluated by histopathology (H&E, TEM), flow cytometry for human immune phenotypes, multiplex ELISA (80 human proteins; 10 injury biomarkers), bulk RNA sequencing, and targeted qPCR. Pearson correlations identified predictors of histopathological injury.

ResultsRenal vasculitis and interstitial nephritis were observed only in ICI-treated HIS-BRGS mice. These kidneys showed a shift toward CD4+ T-cell enrichment with an increased TNF- production capacity compared to CD8+ counterparts. Toxicity was accompanied by increased renal concentrations of human cytokines, chemokines, and soluble receptors. ICI treatment significantly elevated serine proteases (Granzyme A/B) and NGF-{beta}, while decreasing IL-4. Interstitial nephritis correlated with renal PD-1 and MIF. Renal vasculitis correlated with kidney PD-1, CCL1, MIF, Granzyme A, IL-15, and BAFF. Traditional injury biomarkers (KIM-1, NGAL) remained unchanged; however, a trending decrease in EGF was observed.

ConclusionsOur study suggests that shifts in human T-cell populations and specific immune proteins could serve as promising biomarkers and mechanistic targets for ICI nephrotoxicity. The tumor-bearing HIS-BRGS mouse model reproducibly recapitulates the histopathological and immunological features of human ICI-induced nephrotoxicity and represents a validated preclinical platform for testing novel therapeutic interventions to preserve kidney function during cancer immunotherapy.

Translational StatementImmune checkpoint inhibitor (ICI)-associated nephrotoxicity occurs in up to 25% of treated patients, yet the immunological mechanisms driving renal injury remain poorly characterized due to the scarcity of human biopsy material and the absence of robust preclinical models that recapitulate human immune responses. This study demonstrates that tumor-bearing humanized immune system (HIS) mice treated with combined nivolumab and ipilimumab reproducibly develop renal vasculitis and interstitial nephritis mediated by a human CD4+ T cell-dominant infiltrate, mirroring the clinicopathological features reported in patients with ICI-associated acute kidney injury. By integrating histopathology, flow cytometry, multiplex proteomics, and transcriptomics, we identify a coordinated immune network, including IL-15, CCL1, MIF, GZMA, and BAFF, that correlates with the severity of renal pathology and represents tractable mechanistic targets and candidate biomarkers. These findings provide a validated preclinical platform for dissecting irAE mechanisms and testing novel therapeutic strategies to preserve kidney function during cancer immunotherapy.
]]></description>
<dc:creator><![CDATA[ Asby, S., Wen, X., Goedken, M., Ames, B., Shams, S., Thompson, L., Lanis, J., Kostka-Newman, Z., Larsen, K., Tilden, S., Lang, J., Aleksunes, L., Joy, M. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723340</dc:identifier>
<dc:title><![CDATA[Nephrotoxicity of Immune Checkpoint Inhibitors in Mice with a Human Immune System]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723528v1?rss=1">
<title>
<![CDATA[
Systematic toxicological study of PFOS/PFOA co-exposure driving prostate cancer: Core target identification, TME immune remodeling, and combination drug prediction 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723528v1?rss=1
</link>
<description><![CDATA[
BackgroundPer- and polyfluoroalkyl substances (PFAS), particularly perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), are persistent organic pollutants ubiquitous in the environment. Epidemiological evidence has closely linked them to an elevated risk of prostate cancer (PCa). However, the precise molecular mechanisms by which combined PFOS/PFOA exposure promotes prostate cancer and their dynamic effects on the tumor microenvironment remain unclear.

MethodsThis study constructed a multi-module analytical framework integrating network pharmacology and computational biology: (1) Through ADMET toxicity prediction, multi-database target collection (three-way Venn analysis), panoramic GO/KEGG enrichment, focused androgen receptor (AR) axis analysis, GWAS genetic association validation, protein-protein interaction (PPI) network construction, machine learning-based independent screening, and a relaxed intersection strategy, we systematically identified PFOS/PFOA-prostate cancer core targets. (2) Subsequently, a PFAS-PTS score weighted purely by Cox coefficients was employed to drive gene set variation analysis (GSVA)-based pathway enrichment, tumor microenvironment (TME) deconvolution, ordinary differential equation (ODE)-based kinetic modeling, and drug intervention prediction.

ResultsTarget collection identified 100 shared PFOS/PFOA-prostate cancer targets, from which 18 core targets were determined after multi-module screening. These targets were significantly enriched in the AR signaling axis, the PI3K-AKT pathway, and cell cycle regulation. Molecular docking confirmed strong binding affinities of PFOS/PFOA with AR (-9.49/-8.56 kcal/mol), AKT1 (-7.56/-6.93 kcal/mol), and PTEN (-6.36/-6.08 kcal/mol). GSVA revealed that the G2M checkpoint and E2F target gene pathways were significantly upregulated in the high-risk group (padj < 0.001), whereas the androgen response pathway was downregulated (padj = 4.8e-4). TME deconvolution (GSE141445, NNLS) revealed a significantly increased proportion of tumor cells (PCa) (p = 2.4e-4) and markedly reduced CD8+ T cell infiltration (p = 5.7e-4) in the high-risk group, indicating immunosuppressive microenvironment remodeling. ODE-based kinetic modeling confirmed that PFAS promoted tumor cell proliferation and suppressed immune surveillance in a dose-dependent manner. Drug intervention simulation demonstrated that the combination of enzalutamide and Alpelisib achieved optimal tumor cell inhibition (33.9% predicted by the ODE model).

ConclusionPFOS/PFOA promote prostate cancer progression primarily through multi-target synergy involving AR axis disruption, PI3K-AKT pathway activation, and cell cycle dysregulation, while reshaping an immunosuppressive tumor microenvironment. The integrative computational framework established in this study provides systematic computational evidence for risk assessment and therapeutic intervention in PFAS-associated prostate cancer.
]]></description>
<dc:creator><![CDATA[ PAN, J., ZHANG, Y., YANG, A., JIANG, L., SHEN, Y., SUN, Y., ZHU, J., FAN, M., SHI, J. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723528</dc:identifier>
<dc:title><![CDATA[Systematic toxicological study of PFOS/PFOA co-exposure driving prostate cancer: Core target identification, TME immune remodeling, and combination drug prediction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.06.722533v1?rss=1">
<title>
<![CDATA[
Reversible in vivo regulation of drug metabolizing enzyme CYP1A2 activity through a dTAG knock-in strategy 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.06.722533v1?rss=1
</link>
<description><![CDATA[
Drug-metabolizing enzymes determine therapeutic exposure, efficacy and toxicity, but defining their isoform-specific functions in vivo remains challenging. Cytochrome P450 enzymes (P450s) are central to drug metabolism and pharmacokinetics (DMPK) and mediate the phase I metabolism of [~]75% of all marketed drugs. However, conventional knockout models can induce develop-mental and compensatory adaptations, and selective inhibitors are unavailable for many P450 isoforms. Here, we report the use of an inducible chemical-genetic platform for acute and specific degradation of the endogenous P450 enzyme Cyp1a2 in mice. Using CRISPR-Cas9-mediated knock-in editing, we introduced an FKBP12F36V degron into the endogenous Cyp1a2 locus to generate Cyp1a2dTAG mice. Treatment with the dTAG degrader dTAG-13 recruited an E3 ubiquitin ligase to CYP1A2dTAG, resulting in rapid and reversible proteasomal depletion of CYP1A2dTAG in vivo. Temporally controlled CYP1A2dTAG loss altered caffeine pharmacokinetics as expected, validating this model as a functional tool for DMPK studies. By enabling reversible suppression of drug-metabolizing enzymes without permanent deletion or chronic inhibitor exposure, this work establishes targeted protein degradation as a broadly adaptable strategy for studying drug metabolism in vivo and provides a foundation for extending inducible DMPK control to other P450s, conjugating enzymes and transporters.
]]></description>
<dc:creator><![CDATA[ Zhou, S., Ji, X., Li, H., Lanza, D. G., Jung, S. Y., Liu, J., Dogra, A., Nabet, B., MacKenzie, K. R., Wang, J., Matzuk, M. M., Li, F. ]]></dc:creator>
<dc:date>2026-05-12</dc:date>
<dc:identifier>doi:10.64898/2026.05.06.722533</dc:identifier>
<dc:title><![CDATA[Reversible in vivo regulation of drug metabolizing enzyme CYP1A2 activity through a dTAG knock-in strategy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723476v1?rss=1">
<title>
<![CDATA[
Railway Catenary Sparking as a Source of Toxic Copper Ultrafine Particles: Evidence from Realistic In Vitro Inhalation Exposure 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723476v1?rss=1
</link>
<description><![CDATA[
Railway catenary sparking generates airborne ultrafine particles (UFPs) that may pose health risks due to their metallic composition and ability to penetrate deep into the alveolar region of the lungs. Copper, widely used in wires and pantographs, is a major component of these emissions, making copper-rich particles common in railway environments such as subways. However, exposure levels and health impacts remain poorly characterized, and localized hotspots may represent an underrecognized risk in densely populated areas. This study investigated the toxicity of copper UFPs under realistic dosimetry and deposition conditions. Copper UFPs were generated using a spark discharge generator and applied to two in vitro lung models: a 3D co-culture of Calu-3 epithelial cells, THP-1-derived macrophages, and EA.hy926 endothelial cells, and a monoculture of A549 alveolar epithelial cells. Cells were exposed at the air-liquid interface (ALI) using an automated platform to mimic inhalation exposure and UFPs deposition. Copper deposition ranged from 6.5 to 41 ng/cm2, within occupationally relevant levels. A549 cells showed cytotoxic responses consistent with previous studies, whereas the 3D co-culture model revealed broader adverse effects, including inflammation, impaired epithelial barrier integrity, oxidative stress, and early DNA damage. Inflammatory activation also differed between models: A549 cells mainly exhibited transcriptional responses, while the 3D model showed significant secretion of IL-6 and IL-8, associated with interferon signaling. These findings highlight the potential health risks of copper UFPs from railway systems and emphasize the need for improved characterization of UFP exposure in environmental and occupational railway settings.
]]></description>
<dc:creator><![CDATA[ Becker, J., Pantzke, J., Offer, S., Das, A., Mudan, A., Neukirchen, C., Streibel, T., Adam, T., Sklorz, M., Di Bucchianico, S., Zimmermann, R. ]]></dc:creator>
<dc:date>2026-05-11</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723476</dc:identifier>
<dc:title><![CDATA[Railway Catenary Sparking as a Source of Toxic Copper Ultrafine Particles: Evidence from Realistic In Vitro Inhalation Exposure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723338v1?rss=1">
<title>
<![CDATA[
Sex-specific amplification of IKr-blocker-induced action potential prolongation by reduced female IKs repolarization reserve: a computational study using the O'Hara-Rudy human ventricular model 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723338v1?rss=1
</link>
<description><![CDATA[
BackgroundWomen experience drug-induced Torsades de Pointes (TdP) at approximately twice the rate of men across more than 50 QT-prolonging drug classes, yet the quantitative ionic basis of this sex disparity remains incompletely characterised. The slow delayed rectifier current (IKs) is reduced by [~]45% in female compared with male human ventricular cardiomyocytes, reducing the repolarization reserve available to compensate pharmacological IKr block.

MethodsWe implemented the OHara-Rudy (ORd) 2011 undiseased human ventricular epicardial action potential model in Python and parameterised sex variants using the most robustly established human ionic difference: GKs reduced by 45% in females [Kurokawa et al., 2016]. We simulated graded IKr blockade (0-95% in steps of 5%) at three physiologically relevant pacing rates (2 Hz, 1 Hz, 0.5 Hz) after 60 beats of warm-up to approach electrophysiological steady state. Action potential duration at 90% repolarization (APD90), triangulation (APD90-APD30), and repolarization failure (defined as APD90 > 500 ms, a conservative cellular risk marker informed by clinical QTc safety thresholds, or failure to repolarize within the cycle length) were quantified. All simulations used SciPys Radau solver (rtol = 10-, atol = 10-8) with a Numba-JIT-compiled right-hand side for computational efficiency.

ResultsAt baseline (0% block), the female model exhibited longer APD90 than the male at all pacing rates (+2.8 ms at 2 Hz; +4.6 ms at 1 Hz; +4.6 ms at 0.5 Hz). Under progressive IKr blockade, the absolute sex difference in APD90 amplified non-linearly: at 85% block and 1 Hz pacing the female APD90 exceeded the male by 60.4 ms (versus 4.6 ms at baseline; 13-fold amplification). At slow pacing (0.5 Hz), the sex gap was most pronounced: at 85% block, female APD90 was 1127 ms versus 939 ms for the male (+188 ms; 20% more prolonged). The critical APD threshold (>500 ms) was reached by female cells at 5 percentage points lower IKr block than male cells at 1 Hz pacing (55% vs. 60% block), both reported at the first simulated 5%-grid block level exceeding the criterion. Repolarization failure occurred 5 percentage points earlier in females at 1 Hz (90% vs. 95% block). Action potential triangulation was consistently greater in the female model at all block levels and pacing rates.

ConclusionA 45% reduction in IKs conductance is sufficient in this model to produce measurably greater APD90 prolongation under IKr blockade across all tested pacing rates. The non-linear amplification of the sex gap is consistent with the hypothesis that reduced IKs repolarization reserve contributes to greater female susceptibility to drug-induced QT prolongation, and supports testing sex-specific parameterizations in CiPA-style in silico cardiac safety workflows.
]]></description>
<dc:creator><![CDATA[ Magesh Raghavan, T. A. ]]></dc:creator>
<dc:date>2026-05-11</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723338</dc:identifier>
<dc:title><![CDATA[Sex-specific amplification of IKr-blocker-induced action potential prolongation by reduced female IKs repolarization reserve: a computational study using the O'Hara-Rudy human ventricular model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
