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Updated: Dec 28, 2025
Yeast As a Chassis for Developing Functional Assays to Study Human P53
Published on: August 4, 2019
Bifunctional ligand design for modulating mutant p53 aggregation in cancer
Jessica J Miller1, Anaïs Blanchet2, Christophe Orvain2
1Department of Chemistry , Simon Fraser University , Burnaby , British Columbia V5A 1S6 , Canada .
Abstract:
Protein misfolding and aggregation contributes to the development of a wide range of diseases. In cancer, over 50% of diagnoses are attributed to p53 malfunction due to missense mutations, many of which result in protein misfolding and accelerated aggregation. p53 mutations also frequently result in alteration or loss of zinc at the DNA-binding site, which increases aggregation via nucleation with zinc-bound p53. Herein, we designed two novel bifunctional ligands, L and L , to modulate mutant p53 aggregation and restore zinc binding using a metallochaperone approach. Interestingly, only the incorporation of iodine function in L resulted in modulation of mutant p53 aggregation, both in recombinant and cellular environments. Native mass spectrometry shows a protein-ligand interaction for L , as opposed to L , which is hypothesized to lead to the distinct difference in the p53 aggregation profile for the two ligands. Incorporation of a di-2-picolylamine binding unit into the ligand design provided efficient intracellular zinc uptake, resulting in metallochaperone capability for both L and L . The ability of L to reduce mutant p53 aggregation results in increased restoration of p53 transcriptional function and mediates both caspase-dependent and -independent cell death pathways. We further demonstrate that L exhibits minimal toxicity in non-cancerous organoids, and that it is well tolerated in mice. These results demonstrate that iodination of our ligand framework restores p53 function by interacting with and inhibiting mutant p53 aggregation and highlights L as a suitable candidate for comprehensive in vivo anticancer preclinical evaluations.
Insights
Researchers developed a novel ligand (L) that reduces mutant p53 aggregation, restoring its function and mediating cancer cell death. This promising compound shows minimal toxicity and is a candidate for preclinical anticancer evaluations.
Area of Science:
- Biochemistry
- Molecular Biology
- Medicinal Chemistry
Background:
- Protein misfolding and aggregation are implicated in numerous diseases, including cancer.
- Mutant p53 protein, found in over 50% of cancer diagnoses, often misfolds, aggregates, and loses zinc, promoting tumorigenesis.
- Metallochaperone strategies offer a potential therapeutic approach to address mutant p53 dysfunction.
Purpose of the Study:
- To design and evaluate novel bifunctional ligands for modulating mutant p53 aggregation and restoring zinc binding.
- To investigate the role of ligand structure, specifically iodine incorporation, in p53 aggregation modulation.
- To assess the therapeutic potential of the developed ligands as anticancer agents.
Main Methods:
- Design and synthesis of two novel bifunctional ligands (L and L) incorporating a di-2-picolylamine unit for zinc uptake.
- Utilizing native mass spectrometry to analyze protein-ligand interactions.
- Assessing ligand efficacy in modulating mutant p53 aggregation in recombinant and cellular models.
- Evaluating restoration of p53 transcriptional activity and induction of cancer cell death.
- Testing ligand toxicity in non-cancerous organoids and in vivo mouse models.
Main Results:
- Ligand L, containing iodine, effectively modulated mutant p53 aggregation in both recombinant and cellular environments, unlike L.
- Native mass spectrometry confirmed protein-ligand interaction for L, correlating with its aggregation modulation.
- Both ligands demonstrated metallochaperone capability, facilitating intracellular zinc uptake.
- L treatment reduced mutant p53 aggregation, restored p53 transcriptional function, and induced cancer cell death via caspase-dependent and -independent pathways.
- L exhibited minimal toxicity in organoids and was well-tolerated in mice.
Conclusions:
- Iodination of the ligand framework is crucial for modulating mutant p53 aggregation.
- Ligand L acts as a metallochaperone, inhibiting mutant p53 aggregation and restoring tumor suppressor function.
- Ligand L shows significant promise as an anticancer therapeutic candidate for further preclinical evaluation.
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