Abstract A013: Microenvironment driven dynamic deposition of histone H3.3 controls entry and exit from dormancy in disseminated cancer cells
Authors: Drapela Stanislav , Megino-Luque Cristina , Rad Rojan Chimeh , Raizada Devesh , Sarigul Nadir , Ilter Didem , Czerniecki Brian J. , Bravo-Cordero Jose Javier , Gomes Ana P. 
Published: November 2024

Breast cancer metastases can go undetected for long periods of time, are highly resistant to therapy, and are the cause for more than 90% of breast cancer mortality. Dormancy is a critical step in the metastatic cascade, whereby disseminated cancer cells (DCCs) enter a non-proliferative state at distal sites until they reactivate and give rise to overt metastasis, often years after disease remission. As such, dormancy represents a major clinical obstacle in breast cancer patient care. Chromatin remodeling serves as a foundation for the cellular reprogramming required to drive cell fate transitions. However, little is known about the nature of this remodeling in dormancy. Here, we hypothesize that differential deposition of histone H3.3 determines whether DCCs enter and remain dormant in secondary organs or proliferate and form overt metastases. We used a combination of H3.3 deposition sensors and genetic tools to manipulate H3.3 and HIRA in vitro and in vivo to track H3.3 deposition through the metastatic cascade and evaluate its importance for dormancy. To evaluate relevance to patient disease we developed a dormancy score to implement from publicly available RNA seq. To define mechanism of regulation, we used a combination of whole genome sequencing (ChIP-seq, ATAC-seq and RNA-seq) followed by in-depth analysis and functional validation. Our data demonstrates that H3.3 incorporation at secondary sites is heterogenous, revealing a subset of DCCs with low H3.3 in chromatin. Blocking H3.3 deposition onto chromatin leads to a reversible arrest in G0/G1 which tracks with the activation of canonical dormancy signaling. Mechanistically, our data supports a model where blocking H3.3 incorporation into chromatin directly represses the expression of SKP2, the main regulator of progression through the restriction point of cell cycle via regulation of p27 degradation. SKP2 suppression in turn leads to the accumulation of p27 thereby causing cell cycle arrest in G0/G1 and dormancy establishment. We traced this H3.3-mediated regulation of dormancy through the dynamic regulation of H3.3 histone chaperone, HIRA, whose levels inversely correlate with dormancy in experimental models as well as in patient samples. We found that HIRA levels respond to the metastatic microenvironment where fibrotic conditions induce HIRA expression and trigger metastatic outgrowth in an H3.3-dependent manner. Together, our work unveils for the first time a patient-relevant epigenetic mechanism, impinging on H3.3 deposition, as a major regulatory point of DCCs’ cell fate at secondary organs controlling both entry of DCC’s into dormant state as well as their reactivation and thriving as overt metastasis.

Citation Format: Stanislav Drapela, Cristina Megino-Luque, Rojan Chimeh Rad, Devesh Raizada, Nadir Sarigul, Didem Ilter, Brian J. Czerniecki, Jose Javier Bravo-Cordero, Ana P. Gomes. Microenvironment driven dynamic deposition of histone H3.3 controls entry and exit from dormancy in disseminated cancer cells [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr A013.