Biology subjects

Lindner, A. U.

Publications and source records attributed to Lindner, A. U..

3 recordsLinked to original sources

bioRxiv2026

NLRP3 activators disrupt the endocytic AP2 complex and plasma membrane signaling

Organellar perturbations are linked to NLRP3 inflammasome activation, however, it remains unclear whether unrelated agonists converge on a common upstream pathway. Here, we traced intracellular organelle and protein movements by differential ultracentrifugation combined with mass spectrometry-based proteomics. We show that NLRP3 activators uniformly disrupt the endocytic Adaptor Protein 2 (AP2) complex, whereas other subcellular rearrangements are stimulus-specific. We discovered Dynasore as a K-efflux-independent NLRP3 activator that engages this signaling node irrespective of endocytosis inhibition. Pharmacological and genetic perturbation of AP2 renders cells unresponsive to extracellular cues, blunting GPCR signaling, cAMP production, and chemotaxis, thereby enforcing a frozen signaling state that propagates NLRP3 inflammasome activation. Collectively, our study reveals a common surveillance checkpoint linking impaired plasma membrane signaling to the execution of inflammation and cell death. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/707400v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@178eb1dorg.highwire.dtl.DTLVardef@193cb02org.highwire.dtl.DTLVardef@1f50080org.highwire.dtl.DTLVardef@1f3e3e4_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

bioRxiv2022

Bcl-xL is a key mediator of apoptosis following KRASG12C inhibition in KRASG12C mutant colorectal cancer.

PurposeNovel covalent inhibitors of KRASG12C have shown limited response rates in KRASG12C mutant (MT) colorectal cancer (CRC) patients. Thus, novel KRASG12C inhibitor combination strategies that can achieve deep and durable responses are needed. Experimental designSmall molecule KRASG12C inhibitors AZ1569 and AZ8037 were employed. To identify novel candidate combination strategies for AZ1569, we performed RNA sequencing, siRNA and high-throughput drug screening. Top hits were validated in a panel of KRASG12CMT CRC cells and in vivo. AZ1569-resistant CRC cells were generated and characterised. ResultsResponse to AZ1569 was heterogeneous across the KRASG12CMT models. AZ1569 was ineffective at inducing apoptosis when used as single-agent or combined with chemotherapy or agents targeting the EGFR/KRAS/AKT axis. Using a systems biology approach, we identified the anti-apoptotic BH3-family member BCL2L1/Bcl-xL as a top hit mediating resistance to AZ1569. Further analyses identified acute increases in the pro-apoptotic protein BIM following AZ1569 treatment. ABT-263 (Navitoclax), a pharmacological Bcl-2 family-inhibitor that blocks the ability of Bcl-xL to bind and inhibit BIM, led to dramatic and universal apoptosis when combined with AZ1569. Furthermore, this combination also resulted in dramatically attenuated tumour growth in KRASG12CMT xenografts. Finally, AZ1569-resistant cells showed amplification of KRASG12C, EphA2/c-MET activation, increased pro-inflammatory chemokine profile and cross-resistance to several targeted agents. Importantly, KRAS amplification and AZ1569-resistance were reversible upon drug withdrawal, arguing strongly for the use of drug holidays in the case of KRAS amplification. ConclusionsCombinatorial targeting of Bcl-xL and KRASG12C is highly effective, suggesting a novel therapeutic strategy for KRAS G12CMT CRC patients.

cancer biology↗

bioRxiv2021

An atlas of inter- and intra-tumor heterogeneityof apoptosis competency in colorectal cancertissue at single cell resolution

Cancer cells ability to inhibit apoptosis is key to malignant transformation and limits response to therapy. Here, we performed multiplexed immunofluorescence analysis on tissue microarrays with 373 cores from 168 patients, segmentation of 2.4 million individual cells and quantification of 20 cell lineage and apoptosis proteins. Ordinary differential equation-based modelling of apoptosis sensitivity at single cell resolution was conducted and an atlas of inter- and intra-tumor heterogeneity in apoptosis susceptibility generated. We identified an enrichment for BCL2 in immune, and BAK, SMAC and XIAP in cancer cells. ODE-based modelling at single cell resolution identified an enhanced sensitivity of cancer cells to mitochondrial permeabilization and executioner caspase activation compared to immune and stromal cells, with significant inter- and intra-tumor heterogeneity. However, we did not find increased spatial heterogeneity of apoptosis signaling in cancer cells, suggesting that such heterogeneity is an intrinsic, non-genomic property not increased by the process of malignant transformation.

cancer biology↗