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Study breakdown

Cell-Penetrating Stitched Peptides Designed to Block a Cancer-Promoting Protein Called DAXX

evidence
The takeaway

Researchers created stitched peptides that enter cells and inhibit the cancer-linked DAXX protein with higher affinity than its natural binding partners, maintaining nanomolar concentrations inside cells for over 24 hours.

Nanomolar levels maintained 24+ hours

The stitched peptides entered cells and maintained nanomolar intracellular concentrations for at least 24 hours — a critical benchmark for therapeutic utility — without disrupting cell membranes.

What the researchers found

Novel stapled/stitched peptides were designed to target the N-terminal helical bundle of the DAXX oncoprotein, binding with higher affinity than DAXX's known natural interaction partners (Rassf1C, p53, Mdm2, ATRX). The peptides effectively inhibited DAXX and released its auto-inhibited SUMO interaction motif, enabling it to bind SUMO-1. Critically, the stitched peptides entered cells and maintained nanomolar intracellular concentrations for at least 24 hours without disrupting cell membranes.

Why it matters

DAXX is upregulated in many common cancers and its suppression reduces tumor progression, but it has been considered difficult to drug because its critical interactions occur inside cells. These stitched peptides are the first reported cell-permeable inhibitors targeting DAXX, demonstrating that this oncoprotein is druggable and opening a new avenue for cancer therapy development.

How the study worked

Using structural information about the DAXX N-terminal helical bundle domain, the researchers designed stapled and stitched peptide variants targeting the surface where multiple protein partners bind. Binding affinity was measured and compared to known DAXX interaction partners. The effect on DAXX's auto-inhibited SUMO interaction motif was assessed. Cell permeability was evaluated by measuring intracellular peptide concentrations over 24 hours, and membrane integrity was confirmed to rule out non-specific membrane disruption.

Who was studied

In vitro biochemical and cell-based experiments — no human or animal subjects

What this study cannot tell us

This is an early-stage drug development study focused on in vitro characterization — no in vivo animal models or efficacy data were included. While the peptides enter cells at nanomolar levels, their ability to suppress DAXX-dependent tumor growth in living organisms has not been demonstrated. The therapeutic window between DAXX inhibition and normal cellular function is not yet characterized. Specific cancer cell line data and dose-response relationships are not reported in the abstract.

How to read the evidence

This is a preclinical proof-of-concept study demonstrating in vitro binding, functional inhibition, and cell permeability of novel peptides. While the biochemical data is strong, there is no in vivo efficacy data or toxicity profiling, placing this firmly in the early discovery stage.

When this study was published

Published in 2023, this is a recent contribution to the growing field of stitched/stapled peptide therapeutics. DAXX-targeting peptides are at an early stage of development with no clinical candidates yet.

The bigger picture

Many cancer-driving proteins like DAXX have been considered 'undruggable' because they function inside cells and interact through broad protein surfaces rather than small pockets that traditional drugs can fill. Stitched peptides — an evolution of stapled peptides with additional cross-links for stability — represent a growing class of molecules that can tackle these challenging targets. This work adds DAXX to the list of oncoproteins that may be addressable through peptide-based approaches.

Questions still open

  • Can these stitched peptides suppress DAXX-dependent tumor growth in animal models of cancer?
  • What is the selectivity of these peptides for DAXX over other structurally similar proteins, and are there off-target effects?
  • Could these peptides be optimized for oral bioavailability or must they be delivered by injection?

Common questions

What makes a peptide 'stitched' and why does it matter?
A stitched peptide has multiple chemical cross-links along its backbone that lock it into a helical shape, making it more stable than regular peptides. This is an evolution of 'stapled' peptides (which have one cross-link). The extra reinforcement helps the peptide resist breakdown by enzymes, maintain its 3D shape for target binding, and — importantly — cross cell membranes to reach targets inside cells.
Why is the DAXX protein important in cancer?
DAXX is overproduced in many common cancers and plays roles in several processes that help tumors grow, including controlling how DNA is packaged and repaired. When researchers suppress DAXX in cancer cells, tumor progression slows. However, because DAXX works inside cells through protein-protein interactions, it has been difficult to target with conventional drugs — until now.

Read the original research

Stitched peptides as potential cell permeable inhibitors of oncogenic DAXX protein.

RSC chemical biology, 4(12), 1096-1110

Citation

Jelinska, Clare; Kannan, Srinivasaraghavan; Frosi, Yuri; Ramlan, Siti Radhiah; Winnerdy, Fernaldo; Lakshminarayanan, Rajamani; Johannes, Charles W; Brown, Christopher J; Phan, Anh-Tuan; Rhodes, Daniela; Verma, Chandra S. (2023). Stitched peptides as potential cell permeable inhibitors of oncogenic DAXX protein.. RSC chemical biology, 4(12), 1096-1110. https://doi.org/10.1039/d3cb00149k