BML-265 and Tyrphostin AG1478 Disperse the Golgi Apparatus and Abolish Protein Transport in Human Cells

Abstract

The steady-state localization of Golgi-resident glycosylation enzymes in the Golgi apparatus depends on a balance between anterograde and retrograde transport. Using the Retention Using Selective Hooks (RUSH) assay and high-content screening, we identified small molecules that perturb the localization of Mannosidase II (ManII) used as a model cargo for Golgi resident enzymes. In particular, we found that two compounds known as EGFR tyrosine kinase inhibitors, namely BML-265 and Tyrphostin AG1478 disrupt Golgi integrity and abolish secretory protein transport of diverse cargos, thus inducing brefeldin A-like effects. Interestingly, BML-265 and Tyrphostin AG1478 affect Golgi integrity and transport in human cells but not in rodent cells. The effects of BML-265 are reversible since Golgi integrity and protein transport are quickly restored upon washout of the compounds. BML-265 and Tyrphostin AG1478 do not lead to endosomal tubulation suggesting that, contrary to brefeldin A, they do not target the trans-Golgi ARF GEF BIG1 and BIG2. They quickly induce COPI dissociation from Golgi membranes suggesting that, in addition to EGFR kinase, the cis-Golgi ARF GEF GBF1 might also be a target of these molecules. Accordingly, overexpression of GBF1 prevents the effects of BML-265 and Tyrphostin AG1478 on Golgi integrity.

Clinical, radiological and molecular studies in 24 individuals with Dyggve-Melchior-Clausen dysplasia and Smith-McCort dysplasia from India

“…A wider understanding of the complex structure and functioning of the Golgi apparatus is now emerging, enabling better knowledge of Golgi-related disorders with the possibility of targeted treatment options. Two recently described molecules (BML-265 and tyrphostin AG1478) are EGFR (Epidermal Growth Factor Receptor) tyrosine kinase inhibitors, targeting protein transport in human Golgi cells and are postulated to be useful in the therapy of some cancers 47. Similarly pharmacological modulators of Golgi complex functioning (GSK3β inhibitors) for neurological disorders like Alzheimer’s and Parkinson’s disease have recently been evaluated 48…”

Repurposable drugs for SARS-CoV-2 and influenza sepsis with scRNA-seq data targeting post-transcription modifications

“…48 AG-1478 exhibits BFA-like activity by inducing reversible Golgi disruption and inhibiting the secretory transport in human cells. 47 Therefore, AG-1478 and BFA potentially provide a new way to treat COVID-19.…”

“…These drug candidates may function at inhibiting viral replication and spread through a number of mechanisms, including targeting nucleic acid metabolisms and protein synthesis of pathogens as well as the transcription, translation, and post-translational modifications (PTM) of the host cytokines. 47 , 48 Our findings may aid in the rapid preclinical and clinical evaluation of these therapeutics and provide an important drug discovery pipeline to accelerate and facilitate the development of potential treatments for COVID-19 and other viral infections as well as the associated sepsis.…”

Golgi quality control and autophagy

“…Depolymerizes microtubules 22 Vinblastine Depolymerizes microtubules 23 Vincristine Depolymerizes microtubules 23 Vindesine Depolymerizes microtubules 23 Vinorelbine Depolymerizes microtubules 23 Others 2-PB Promotes Golgi proteins degradation 24 AMF26 Inhibits Arf1-ArfGEF interaction 25 Amphotericin B1 Reduces Golgi PI(4)P 26 BML-265 An EGFR inhibitor 27 Brefeldin A Binds to ARF-ADP/GEF complex; blocks GDP/GTP exchange 28 CBM Inhibits coatomer binding to the Golgi membrane 29 EtOH Inhibits ARF1 30…”