bims-momema 2022-03-06 papers

Issue of 2022–03–06
four papers selected by
Harilaos Filippakis, Harvard University

Proc Natl Acad Sci U S A. 2022 Mar 08. 119(10): e2107453119

O-GlcNAcylation regulates epidermal growth factor receptor intracellular trafficking and signaling.

Liming Wu, Yaxian Cheng, Didi Geng, Zhiya Fan, Bingyi Lin, Qiang Zhu, Jingchao Li, Weijie Qin, Wen Yi.

SignificanceEpidermal growth factor receptor (EGFR) is one of the most important membrane receptors that transduce growth signals into cells to sustain cell growth, proliferation, and survival. EGFR signal termination is initiated by EGFR internalization, followed by trafficking through endosomes, and degradation in lysosomes. How this process is regulated is still poorly understood. Here, we show that hepatocyte growth factor regulated tyrosine kinase substrate (HGS), a key protein in the EGFR trafficking pathway, is dynamically modified by a single sugar N-acetylglucosamine. This modification inhibits EGFR trafficking from endosomes to lysosomes, leading to the accumulation of EGFR and prolonged signaling. This study provides an important insight into diseases with aberrant growth factor signaling, such as cancer, obesity, and diabetes.

Keywords: EGFR; O-GlcNAcylation; endosomal sorting; membrane receptors

DOI: https://doi.org/10.1073/pnas.2107453119

Cell Death Dis. 2022 Mar 02. 13(3): 199

Extracellular ATP promotes breast cancer chemoresistance via HIF-1α signaling.

Hui Yang, Yue-Hang Geng, Peng Wang, Hong-Quan Zhang, Wei-Gang Fang, Xin-Xia Tian.

We have previously demonstrated that extracellular adenosine 5'-triphosphate (ATP) promotes breast cancer cell chemoresistance. However, the underlying mechanism remains unclear. Using a cDNA microarray, we demonstrated that extracellular ATP can stimulate hypoxia-inducible factor (HIF) signaling. In this study, we report that hypoxia-inducible factor 1α (HIF-1α) was upregulated after ATP treatment and mediated the ATP-driven chemoresistance process. We aimed to investigate the mechanisms and identify potential clinically relevant targets that are involved. Using mass spectrometry, we found that aldolase A (ALDOA) interacts with HIF-1α and increases HIF-1α expression. We then demonstrated that STAT3-ALDOA mediates ATP-HIF-1α signaling and upregulates the HIF-1 target genes adrenomedullin (ADM) and phosphoinositide-dependent kinase-1 (PDK1). Moreover, we show that PI3K/AKT acts upstream of HIF-1α in ATP signaling and contributes to chemoresistance in breast cancer cells. In addition, HIF-1α-knockdown or treatment with direct HIF inhibitors combined with the ATP hydrolase apyrase in MDA-MB-231 cells induced enhanced drug sensitivity in nude BALB/c mice. We then used in vitro spheroid formation assays to demonstrate the significance of ATP-HIF-1α in mediating chemoresistance. Furthermore, considering that indirect HIF inhibitors are effective in clinical cancer therapy, we treated tumor-bearing BALB/c mice with STAT3 and PI3K/AKT inhibitors and found that the dual-targeting strategy sensitized breast cancer to cisplatin. Finally, using breast cancer tissue microarrays, we found that ATP-HIF-1α signaling is associated with cancer progression, poor prognosis, and resistance to chemotherapy. Taken together, we suggest that HIF-1α signaling is vital in ATP-driven chemoresistance and may serve as a potential target for breast cancer therapies.

DOI: https://doi.org/10.1038/s41419-022-04647-6

Proc Natl Acad Sci U S A. 2022 Mar 08. 119(10): e2110415119

Amino acid sensor conserved from bacteria to humans.

Vadim M Gumerov, Ekaterina P Andrianova, Miguel A Matilla, Karen M Page, Elizabet Monteagudo-Cascales, Annette C Dolphin, Tino Krell, Igor B Zhulin.

SignificanceAmino acids are the building blocks of life and important signaling molecules. Despite their common structure, no universal mechanism for amino acid recognition by cellular receptors is currently known. We discovered a simple motif, which binds amino acids in various receptor proteins from all major life-forms. In humans, this motif is found in subunits of calcium channels that are implicated in pain and neurodevelopmental disorders. Our findings suggest that γ-aminobutyric acid-derived drugs bind to the same motif in human proteins that binds natural ligands in bacterial receptors, thus enabling future improvement of important drugs.

Keywords: evolution; gabapentin; ion channels; serine/threonine kinases; signal transduction

DOI: https://doi.org/10.1073/pnas.2110415119

Methods Mol Biol. 2022 ;2419 247-255

Monitoring Modified Lipoprotein Uptake and Macropinocytosis Associated with Macrophage Foam Cell Formation.

Alaa Alahmadi, Dipak P Ramji.

Macrophage foam cell formation plays a crucial role in the initiation and progression of atherosclerosis. Macrophages uptake native and modified low density lipoprotein (LDL) through either receptor-dependent or receptor-independent mechanisms to transform into lipid laden foam cells. Foam cells are involved in the formation of fatty streak that is seen during the early stages of atherosclerosis development and therefore represents a promising therapeutic target. Normal or modified lipoproteins labeled with fluorescent dyes such as 1,1'-dioctadecyl-3-3-3',3'-tetramethylindocarbocyanine perchlorate (Dil) are often used to monitor their internalization during foam cell formation. In addition, the fluorescent dye Lucifer Yellow (LY) is widely used as a marker for macropinocytosis activity. In this chapter, we describe established methods for monitoring modified lipoprotein uptake and macropinocytosis during macrophage foam cell formation.

Keywords: 1′-Dioctadecyl-3-3-3′,3′-tetramethylindocarbocyanine perchlorate dye (Dil); Foam cells; LDL; Lucifer Yellow; Macropinocytosis

DOI: https://doi.org/10.1007/978-1-0716-1924-7_14