Discovery of Novel Proteasome Inhibitors Using a High-Content Cell-Based Screening System

Lavelin, Irina; Beer, Avital; Kam, Zvi; Rotter, Varda; Oren, Moshe; Navon, Ami; Geiger, Benjamin

doi:10.1371/journal.pone.0008503

Cited by 20 publications

(7 citation statements)

References 42 publications

(52 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A recent study [34] has suggested that the sensitivity to proteasome inhibition is associated with the status of p53. However, apoptosis triggered by proteasome inhibition appears to be independent of p53 in prostate cancer [35], multiple myeloma [30], and colon cancer cells [36].…”

Section: Discussionmentioning

confidence: 99%

Targeting Neuroblastoma Stem Cells with Retinoic Acid and Proteasome Inhibitor

Hämmerle

Yáñez

Palanca³

et al. 2013

PLoS ONE

View full text Add to dashboard Cite

BackgroundNeuroblastma cell lines contain a side-population of cells which express stemness markers. These stem-like cells may represent the potential underlying mechanism for resistance to conventional therapy and recurrence of neuroblastoma in patients.Methodology/Principal FindingsTo develop novel strategies for targeting the side-population of neurobastomas, we analyzed the effects of 13-cis-retinoic acid (RA) combined with the proteasome inhibitor MG132. The short-term action of the treatment was compared with effects after a 5-day recovery period during which both chemicals were withdrawn. RA induced growth arrest and differentiation of SH-SY5Y and SK-N-BE(2) neuroblastoma cell lines. Inhibition of the proteasome caused apoptosis in both cell lines, thus, revealing the critical role of this pathway in the regulated degradation of proteins involved in neuroblastoma proliferation and survival. The combination of RA with MG132 induced apoptosis in a dose-dependent manner, in addition to promoting G2/M arrest in treated cultures. Interestingly, expression of stem cell markers such as Nestin, Sox2, and Oct4 were reduced after the recovery period of combined treatment as compared with untreated cells or treated cells with either compound alone. Consistent with this, neurosphere formation was significantly impaired by the combined treatment of RA and MG132.ConclusionsGiven that stem-like cells are associated with resistant to conventional therapy and are thought to be responsible for relapse, our results suggest that dual therapy of RA and proteasome inhibitor might be beneficial for targeting the side-population of cells associated residual disease in high-risk neuroblastoma.

show abstract

Section: Discussionmentioning

confidence: 99%

Targeting Neuroblastoma Stem Cells with Retinoic Acid and Proteasome Inhibitor

Hämmerle

Yáñez

Palanca³

et al. 2013

PLoS ONE

View full text Add to dashboard Cite

show abstract

“…Likewise, TMPRSS4 activates cell-mediated fusion of MERS-CoV and SARS-CoV, but it does not activate infection. Other TTSPs, TMPRSS11a, and TMPRSS11e, activate S-protein fusion of MERS-CoV, while TMPRSS11a activates SARS-CoV fusion S-protein, which causes infection (Tang et al 2020 ; Kam et al 2009 ). Human airway trypsin-like protease (HAT) known as TMPRSS11d, can activate MERS-CoV (Tang et al 2020 ; Kam et al 2009 ; Bertram et al 2011 ).…”

Section: Human Cellular Receptors As Targets Of Coronavirusesmentioning

confidence: 99%

“…Other TTSPs, TMPRSS11a, and TMPRSS11e, activate S-protein fusion of MERS-CoV, while TMPRSS11a activates SARS-CoV fusion S-protein, which causes infection (Tang et al 2020 ; Kam et al 2009 ). Human airway trypsin-like protease (HAT) known as TMPRSS11d, can activate MERS-CoV (Tang et al 2020 ; Kam et al 2009 ; Bertram et al 2011 ). Although, transient expression of TMPRSS2, TMPRSS4, TMPRSS11a, TMPRSS11d, and TMPRSS11e enhances SARS-CoV-2 S-protein-mediated cell–cell fusion, these proteases activating the S-protein for infection are yet to be investigated (Ou et al 2020 ; Tang et al 2020 ).…”

Section: Human Cellular Receptors As Targets Of Coronavirusesmentioning

confidence: 99%

Human cell receptors: potential drug targets to combat COVID-19

Raghav¹,

Kalyanaraman

Kumar

2021

Amino Acids

View full text Add to dashboard Cite

“…For control group, Ad-Red was substituted for TE6 or TE7 adenovirus in the p53 control and Rb control, respectively. 12-24h later, 10 μl cells were collected and plated in 384 well plates containing 10 μl DMEM with 20 μM compounds, making the final screening compound concentration of 10 μM, which is a concentration commonly used in cell-based high-throughput screens [ 13 , 14 ]. Experimental wells were plated with 2000 p53-TE6 cells and 2000 Rb-TE7 cells per well.…”

Section: Methodsmentioning

confidence: 99%

A High-Throughput Cell-Based Screen Identified a 2-[(E)-2-Phenylvinyl]-8-Quinolinol Core Structure That Activates p53

et al. 2016

View full text Add to dashboard Cite

p53 function is frequently inhibited in cancer either through mutations or by increased degradation via MDM2 and/or E6AP E3-ubiquitin ligases. Most agents that restore p53 expression act by binding MDM2 or E6AP to prevent p53 degradation. However, fewer compounds directly bind to and activate p53. Here, we identified compounds that shared a core structure that bound p53, caused nuclear localization of p53 and caused cell death. To identify these compounds, we developed a novel cell-based screen to redirect p53 degradation to the Skip-Cullin-F-box (SCF) ubiquitin ligase complex in cells expressing high levels of p53. In a multiplexed assay, we coupled p53 targeted degradation with Rb1 targeted degradation in order to identify compounds that prevented p53 degradation while not inhibiting degradation through the SCF complex or other proteolytic machinery. High-throughput screening identified several leads that shared a common 2-[(E)-2-phenylvinyl]-8-quinolinol core structure that stabilized p53. Surface plasmon resonance analysis indicated that these compounds bound p53 with a KD of 200 ± 52 nM. Furthermore, these compounds increased p53 nuclear localization and transcription of the p53 target genes PUMA, BAX, p21 and FAS in cancer cells. Although p53-null cells had a 2.5±0.5-fold greater viability compared to p53 wild type cells after treatment with core compounds, loss of p53 did not completely rescue cell viability suggesting that compounds may target both p53-dependent and p53-independent pathways to inhibit cell proliferation. Thus, we present a novel, cell-based high-throughput screen to identify a 2-[(E)-2-phenylvinyl]-8-quinolinol core structure that bound to p53 and increased p53 activity in cancer cells. These compounds may serve as anti-neoplastic agents in part by targeting p53 as well as other potential pathways.

show abstract

Discovery of Novel Proteasome Inhibitors Using a High-Content Cell-Based Screening System

Cited by 20 publications

References 42 publications

Targeting Neuroblastoma Stem Cells with Retinoic Acid and Proteasome Inhibitor

Targeting Neuroblastoma Stem Cells with Retinoic Acid and Proteasome Inhibitor

Human cell receptors: potential drug targets to combat COVID-19

A High-Throughput Cell-Based Screen Identified a 2-[(E)-2-Phenylvinyl]-8-Quinolinol Core Structure That Activates p53

Contact Info

Product

Resources

About