Functional consequences of a p53-MDM2-p21 incoherent feedforward loop

Abstract Genetically identical tumor cells can respond heterogeneously to therapy, with a subpopulation of cells often entering a temporarily arrested treatment-tolerant state before repopulating the tumor. To investigate how heterogeneity in the cell cycle arrest protein p21 arises, we imaged the dynamics of p21 transcription and protein expression along with those of p53, its transcriptional regulator, in single MCF-7 cells using live cell fluorescence microscopy. Using this approach, we found that the rate of p21 transcription depends on the change in p53 rather than its absolute level. Through combined theoretical and experimental modeling, we determined that p21 transcription is governed by an incoherent feedforward loop mediated by MDM2. This network architecture facilitates rapid induction of p21 expression and variability in p21 transcription. Abrogating the feedforward loop overcomes rapid S-phase p21 degradation, with cells transitioning into a quiescent state that transcriptionally resembles a treatment-tolerant persister state. The potential implications of our findings warrant consideration in the design of therapeutic strategies based on activating p53. Competing Interest Statement The authors declare the following competing interests: F.M. is a cofounder and advisor to Harbinger Health, an advisor to Zephyr AI, and serves on the board of directors of Recursion Pharmaceuticals. F.M. declares that none of these relationships are directly or indirectly related to the content of this manuscript. Footnotes ↵11 Lead contact ↵+ Equal contribution. Figures 1, 4, 5 and 6 revised; Supplementary Table 1 revised; Supplementary Figures 3 and 5 added; Supplementary Figures 1, 4, 6 and 7 revised; text revised to include new methods and results, and clarify methodological details and interpretations of results; author added.