Cytoplasmic YAP1‐mediated ESCRT‐III assembly promotes autophagic cell death and is ubiquitinated by NEDD4L in breast cancer

Abstract

Background Nuclear Yes1‐associated transcriptional regulator (YAP1) promotes tumor progression. However, the function of cytoplasmic YAP1 in breast cancer cells and its impact on the survival of breast cancer patients remain unclear. Our research aimed to explore the biological function of cytoplasmic YAP1 in breast cancer cells and the possibility of cytoplasmic YAP1 as a predictive marker of breast cancer survival. Methods We constructed cell mutant models, including NLS‐YAP15SA (nuclear localized), YAP1S94A (incapable of binding to the TEA domain transcription factor family) and YAP1S127D (cytoplasmic localized), and used Cell Counting Kit‐8 (CCK‐8) assays, 5‐ethynyl‐2’‐deoxyuridine (EdU) incorporation assays, and Western blotting (WB) analysis to detect cell proliferation and apoptosis. The specific mechanism of cytoplasmic YAP1‐mediated endosomal sorting complexes required for transport III (ESCRT‐III) assembly was studied by co‐immunoprecipitation, immunofluorescence staining, and WB analysis. Epigallocatechin gallate (EGCG) was used to simulate YAP1 retention in the cytoplasm in in vitro and in vivo experiments to study the function of cytoplasmic YAP1. YAP1 binding to NEDD4‐like E3 ubiquitin protein ligase (NEDD4L) was identified using mass spectrometry and was verified in vitro. Breast tissue microarrays were used to analyze the relationship between cytoplasmic YAP1 expression and the survival of breast cancer patients. Results YAP1 was mainly expressed in the cytoplasm in breast cancer cells. Cytoplasmic YAP1 promoted autophagic death of breast cancer cells. Cytoplasmic YAP1 bound to the ESCRT‐III complex subunits charged multivesicular body protein 2B (CHMP2B) and vacuolar protein sorting 4 homolog B (VPS4B), promoting assembly of CHMP2B‐VPS4B and activating autophagosome formation. EGCG retained YAP1 in the cytoplasm, promoting the assembly of CHMP2B‐VPS4B to promote autophagic death of breast cancer cells. YAP1 bound to NEDD4L, and NEDD4L mediated ubiquitination and degradation of YAP1. Breast tissue microarrays revealed that high levels of cytoplasmic YAP1 were beneficial to the survival of breast cancer patients. Conclusions Cytoplasmic YAP1 mediated autophagic death of breast cancer cells by promoting assembly of the ESCRT‐III complex; furthermore, we established a new breast cancer survival prediction model based on cytoplasmic YAP1 expression.

Identification of a PANoptosis-related Gene Signature for Predicting the Prognosis, Tumor Microenvironment and Therapy Response in Breast Cancer

“…On the other hand, CHMP2B is decreased in patients with endometrial carcinoma and the urinary exosomes of patients with colorectal cancer compared to healthy individuals 57 , 58 . Recently, Guo et al revealed that cytoplasmic Yes1-associated transcriptional regulator (YAP1) could inhibit the proliferation of breast tumors by promoting autophagy, potentially through the combination of CHMP2B and VPS4B 59 . However, the experiments conducted and the model constructed in this research were mainly based on YAP1, while the role of CHMP2B itself remained to be further explored.…”

Ferroptosis-Regulated Natural Products and miRNAs and Their Potential Targeting to Ferroptosis and Exosome Biogenesis

“…Auriculasin [25] Colon Ca ↓ pAIFM1 [25] Bromelain [60] Colon Ca ↑ ATG5, ATG7 [61], ACSL4 [60] Curcumin [62] Colon Ca ↑ ATF3 [63], ACSL4 [64], IREB2 [63], HMOX1 [65] ↓ SLC7A11, GPX4 [62,64], HIF1A [66], NEDD4 [67,68] Dihydroartemisinin [69] Glioma ↑ ATF4 [70,71], NCOA4 [72], HMOX1 [69] ↓ GPX4, SLC7A11, SLC3A2 [70], FTH1 [73] Dihydroisotanshinone I [74] Glioma ↑ ACSL4 [74] ↓ GPX4 [74] Diplacone [75] Lung Ca DMOCPTL [76] Breast Ca ↓ GPX4 [76] Epigallocatechin gallate [77] Pancreatic Ca ↑ ATF4 [78], ATG5, ATG7 [79], YAP1 [80] ↓ SP1, TP53 [81] Epunctanone [82] Leukemia…”

“…For the modulation of ferroptosis-inducing genes, EGCG enhances the ferroptosis of pancreatic cancer cells by promoting the degradation of the ferroptosis-inhibiting GPX4 gene [ 77 ]. For non-ferroptosis responses, EGCG may induce ER stress [ 78 ], autophagy [ 79 , 80 ], and apoptosis [ 79 ]. EGCG triggers the ER stress of colon cancer cells by upregulating the ferroptosis-inducing ATF4 gene [ 78 ].…”

“…EGCG triggers the ER stress of colon cancer cells by upregulating the ferroptosis-inducing ATF4 gene [ 78 ]. EGCG induces the autophagy-mediated death of breast cancer cells by retaining the ferroptosis-inducing YAP1 gene in the cytoplasm [ 80 ]. EGCG induces the antiproliferation, apoptosis, and autophagy of umbilical vein endothelial cells grown on 316L stainless steel by upregulating ferroptosis-inducing genes ( ATG5 and ATG7 ) [ 79 ].…”

NEDD4L in human tumors: regulatory mechanisms and dual effects on anti-tumor and pro-tumor

“…Glutathione peroxidase 4 (GPX4) determines the sensitivity of lung cancer cells to etoposide-induced ferroptosis, and NEDD4L can ubiquitinate GPX4, resulting in its subsequent degradation (Cheng et al, 2023). Cytoplasmic yes-associated protein 1 (YAP1) promotes autophagic death in breast cancer cells, and NEDD4L mediates ubiquitination and downregulation of YAP1 (Guo et al, 2023). Human organic anion transporter 3 (hOAT3) is highly expressed in the kidneys and plays a key role in the secretion of clinically important drugs, including anti-cancer drugs (You, 2002;Erdman et al, 2006).…”