Peptides genetically selected for NF-κB activation cooperate with oncogene Ras and model carcinogenic role of inflammation

Protein-protein interactions (PPIs) are emerging as a promising new class of drug targets. Here, we present a novel high-throughput approach to screen inhibitors of PPIs in cells. We designed a library of 50,000 human peptide binding motifs and used a pooled lentiviral system to express them intracellularly and screen for their effects on cell proliferation. We thereby identified inhibitors that drastically reduced the viability of a pancreas cancer line (RWP1) while leaving a control line virtually unaffected. We identified their target interactions computationally, and validated a subset in experiments. We also discovered their potential mechanisms of action including apoptosis and cell cycle arrest. Finally, we confirmed that synthetic lipopeptide versions of our inhibitors have similarly specific and dosage dependent effects on cancer cell growth. Our screen reveals new drug targets and peptide drug leads and it provides a rich dataset covering phenotypes for inhibition of thousands of interactions.

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“…1 and Online methods). A conceptually related approach was recently published that identifies bioactive extracellular peptides 18 .…”

Section: Resultsmentioning

confidence: 99%

Pooled screening for antiproliferative inhibitors of protein-protein interactions

et al. 2016

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“…Sufficiency of NF-kB activation for overcoming p53's ability to trigger oncogene-induced senescence was shown in our experiments using a set of NF-kB-activating selectable peptides (NASPs) (Natarajan et al 2014). Transduction of NASPs into mouse and rat embryo fibroblasts did not, in itself, alter their growth.…”

Section: Inflammation Can Suppress P53 Functionmentioning

confidence: 63%

p53 and the Carcinogenicity of Chronic Inflammation

Gudkov

Komarova

2016

Cold Spring Harb Perspect Med

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Chronic inflammation is a major cancer predisposition factor. Constitutive activation of the inflammation-driving NF-κB pathway commonly observed in cancer or developed in normal tissues because of persistent infections or endogenous tissue irritating factors, including products of secretion by senescent cells accumulating with age, markedly represses p53 functions. In its turn, p53 acts as a suppressor of inflammation helping to keep it within safe limits. The antagonistic relationship between p53 and NF-κB is controlled by multiple mechanisms and reflects cardinal differences in organismal responses to intrinsic and extrinsic cell stresses driven by these two transcription factors, respectively. This provides an opportunity for developing drugs to treat diseases associated with inappropriate activity of either p53 or NF-κB through targeting the opposing pathway. Several drug candidates of this kind are currently in clinical testing. These include anticancer small molecules capable of simultaneous suppression of p53 and activation of NF-κB and NF-κB-activating biologics that counteract p53-mediated pathologies associated with systemic genotoxic stresses such as acute radiation syndrome and side effects of cancer treatment.

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“…KO experiments of the NF-jB-pathway revealed that NASPs occur upstream of TNF receptorassociated factor 6. After transduction of NASPs into the fibroblasts of mice and rats resulted in the co-expression of Ras (H-Ras V12) and revealed that rodent fibroblasts mastered the p53-dependent senescence which is mediated by H-Ras V12 and this in turn resulted in a transformed carcinogenic phenotype [117]. These observations underly the different functions of NF-jB and its cooperation with Ras as an oncogene by the attenuation of p53 as well its effects on the inflammatory cascade.…”

Section: Nf-jb Signaling and Crosstalk During Carcinogenesis In Cell mentioning

confidence: 97%

“…For example, the chemotherapeutic efficacy of cisplatin can be enhanced by inhibiting NF-jB in vitro and in vivo [116]. In 2014, an in vitro model of NF-jB driven carcinogenesis was published [117]. The authors used a cell-based phenotypic readout and isolated 12 genetic elements that induced NF-jB activity (NF-jB-activating genetic elements, NASPs) of lentiviral libraries encoding 20 or 50 amino acid-long polypeptides "none of which was previously associated with NF-jB activation, were isolated from libraries of 200 000 peptides derived from 500 human extracellular proteins".…”

Section: In Vitro Nf-jb Cancer Modelmentioning

confidence: 99%

“…The jB-motif, by itself, has a certain degree of variability which suggests that it has the ability to fine tune its effects [371]. NF-jB counteracts senescence induced by oncogenes and thus drives pro-carcinogenic inflammation [117,179]. KO experiments of the NF-jB-pathway revealed that NASPs occur upstream of TNF receptorassociated factor 6.…”

Section: Nf-jb Signaling and Crosstalk During Carcinogenesis In Cell mentioning

confidence: 99%

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NF-κB signaling and crosstalk during carcinogenesis

Brücher

Läng

Jamall

2019

4open

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Transcription factors (TFs) are proteins that control the transcription of genetic information from DNA to mRNA by binding to specific DNA sequences either on their own or with other proteins as a complex. TFs thus support or suppress the recruitment of the corresponding RNA polymerase. In general, TFs are classified by structure or function. The TF, Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), is expressed in all cell types and tissues. NF-κB signaling and crosstalk are involved in several steps of carcinogenesis including in sequences involving pathogenic stimulus, chronic inflammation, fibrosis, establishment of its remodeling to the precancerous niche (PCN) and transition of a normal cell to a cancer cell. Triggered by various inflammatory cytokines, NF-κB is activated along with other TFs with subsequent stimulation of cell proliferation and inhibition of apoptosis. The involvement of NF-κB in carcinogenesis provides an opportunity to develop anti-NF-κB therapies. The complexity of these interactions requires that we elucidate those aspects of NF-κB interactions that play a role in carcinogenesis, the sequence of events leading to cancer.

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Peptides genetically selected for NF-κB activation cooperate with oncogene Ras and model carcinogenic role of inflammation

Cited by 15 publications

References 49 publications

Pooled screening for antiproliferative inhibitors of protein-protein interactions

Pooled screening for antiproliferative inhibitors of protein-protein interactions

p53 and the Carcinogenicity of Chronic Inflammation

NF-κB signaling and crosstalk during carcinogenesis

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