Gain-of-function p53 protein transferred via small extracellular vesicles promotes conversion of fibroblasts to a cancer-associated phenotype

Abstract

SUMMARY Tumor and stromal interactions consist of reciprocal signaling through cytokines, growth factors, direct cell-cell interactions, and extracellular vesicles (EVs). Small EVs (≤200 nm) have been considered critical messengers of cellular communication during tumor development. Here, we demonstrate that gain-of-function (GOF) p53 protein can be packaged into small EVs and transferred to fibroblasts. GOF p53 protein is selectively bound by heat shock protein 90 (HSP90), a chaperone protein, and packaged into small EVs. Inhibition of HSP90 activity blocks packaging of GOF, but not wild-type, p53 in small EVs. GOF p53-containing small EVs result in their conversion to cancer-associated fibroblasts. In vivo studies reveal that GOF p53-containing small EVs can enhance tumor growth and promote fibroblast transformation into a cancer-associated phenotype. These findings provide a better understanding of the complex interactions between cancer and stromal cells and may have therapeutic implications.

Comprehensive proteomic profiling of plasma and serum phosphatidylserine-positive extracellular vesicles reveals tissue-specific proteins

“…EVs carry nucleic acids, such as microRNA, mRNA, and noncoding RNA (ncRNA), lipids, and proteins, which can transfer to recipient cells for cell-to-cell communication ( Delpech et al., 2019 ; Hoshino et al., 2020 ; Muraoka et al., 2020b ; You et al., 2020 ). EVs have been extensively investigated for their function and roles in intracellular communication during cancer development and neurodegenerative disease progression ( Hoshino et al., 2015 ; Hosseini et al., 2016 ; Ma et al., 2021 ; Patel and Weaver, 2021 ; Ruan et al., 2020 ; Sun et al., 2021 ).…”

CD63-mediated cloaking of VEGF in small extracellular vesicles contributes to anti-VEGF therapy resistance

“…To demonstrate the presence of VEGF in small EVs, we performed a sucrose/D 2 O cushion ultracentrifugation (SUC) to deplete potential protein contaminations from cell culture supernatants as previously described ( Ma et al, 2021 ).…”

“…TEM was performed by Kenneth Dunner Jr. with the High Resolution Electron Microscopy Facility of MDACC as described previously ( Ma et al, 2021 ).…”

“…Next, the solution was transferred to a fresh polypropylene centrifuge tube (Beckman Coulter) and centrifuged for 2 hr at 100,000 3 g and 4 C (repeated once). After this, the pellet was re-suspended in 20 mL of PBS and stored at À80 C. Purification of small EVs by 30% Sucrose/Deuterium oxide (D 2 O) cushion To demonstrate the presence of VEGF in small EVs, we performed a sucrose/D 2 O cushion ultracentrifugation (SUC) to deplete potential protein contaminations from cell culture supernatants as previously described (Ma et al, 2021). Transmission electron microscopy (TEM) TEM was performed by Kenneth Dunner Jr. with the High Resolution Electron Microscopy Facility of MDACC as described previously (Ma et al, 2021).…”

Regulation of Neuroendocrine-like Differentiation in Prostate Cancer by Non-Coding RNAs

“…The complexity and type of changes that lead to the deregulation of AR, PTEN, p53, and pRB signalling during PCa progression is a matter of intensive investigation and vivid discussions [112][113][114][115][116]. They may encompass processes such as expression of constitutively active AR variants [117], ligand-independent activation of AR [118], or expression of gain-of-function p53 mutants, which have been recently demonstrated to induce conversion of fibroblasts to a cancer-associated phenotype that supports increased tumour growth and metastasis [108,119,120]. Acquisition of the neuroendocrine program is accompanied by genetic alterations and rewiring of other important signalling networks such as those mediated by Aurora kinase A (AURKA) or PI3K/AKT [121][122][123][124].…”