Transcriptional Co-Activator P300 Promotes Extra-Medullary Disease Formation and Myeloma Disease Progression
Authors: Shome Rajib , Talluri Srikanth , Fukushima Tsuyoshi , Samur Mehmet K. , Nakamoto-Matsubara Rie , Panaroni Cristina , Fulzele Keertik , Ochi Kiyosumi , Han Ryan Seongmin , Rexha Farah , Scadden David , Munshi Nikhil C , Raje Noopur S. 
Published: December 2023

The transcription co-activator P300 regulates gene expression by histone H3 lysine 27 acetylation (H3K27ac) of chromatin and transcriptional regulators. P300 has been suggested to control a variety of biological functions, including differentiation, cell-cycle control, proliferation, apoptosis, and DNA damage response. The clinicopathologic/prognostic importance of P300 in multiple myeloma (MM) is unknown to date. P300 has been linked to the induction of epithelial-to-mesenchymal transition (EMT) and metastasis in a variety of solid tumor malignancies. Like EMT in solid tumors, extra-medullary disease (EMD) in MM relates to a poor prognosis and resistance to treatment. Similarly, the molecular changes concerning EMD in MM are poorly understood, and the role of P300 in MM EMD is yet to be investigated.

We have employed chemical and functional genomics to unravel the role of P300 in MM disease progression and EMD formation. Six human MM cell lines-MM1S, KMS-18, H929, U266, OPM-2, and AMO-1-were used to construct lentiviral shRNA-mediated P300 knockdown (K/D) cells. In addition, to analyzing the effect of time-dependent P300 degradation, we have used a proteolysis-targeting chimera (PROTAC) compound termed “JQAD1” that selectively targets P300 for degradation. Treatment with JQAD1 effectively reduced the survival with an IC 50 of 50 nM in MM1S cells treated for 72 h. Furthermore, we wanted to understand the mechanism of P300 degradation-mediated inhibition of cell proliferation. Cell cycle analysis revealed a 40% increase in G0/G1 phase cell cycle arrest following treatment with 50 nM JQAD1 for 72 h. Blocking CD138 receptors is known to enhance MM sensitivity to proteasome inhibitors and significantly reduce tumor size. Immuno-flowcytometry analysis revealed that loss of P300 reduces CD138 expression in MM cells, suggesting their susceptibility to proteasome inhibitors.

Next, we wanted to understand if P300 plays a role in developing EMD in MM. Boyden chamber-based migration and invasion assays exhibited reduced migration and invasion of P300 K/D cells with respect to control cells. Furthermore, to understand the distribution and localization of P300 silenced MM cells in vivo, we have used immune deficient NSG mice and studied MM progression after intravenous tumoral injection. In vivo studies revealed that P300 silencing led to reduced MM liver, kidney homing, and EMD formation, as observed through bioluminescence imaging, histology, and flow cytometry analysis, respectively. Further investigations are currently underway to find novel interaction partners and potential targets of P300 in MM and to delineate its mechanism of action.

In conclusion, our research indicates that P300 is critical for the development of EMD and the progression of MM. This could potentially serve as a new therapeutic target for MM relapse and dissemination.