We’re excited about this direction. Synthetic TF tethering may provide a general approach for engineering gene expression in cells, and a path toward more programmable control of cell identity.
I’ll again thank @igvfconsortium.bsky.social and team. Please reach out with feedback.
My group has long used epigenomics to define cell types. Since joining @harvard.edu SCRB, we set out to create strategies to engineer cell fate and state.
Here, we look to engineer CD8 T cells, where exhaustion limits their broader therapeutic use.
Many regulators of T cell exhaustion have been discovered. As our community saturates discovery of endogenous regulators, we reasoned there may be major opportunities to combine perturbations and/or design synthetic proteins.
We hypothesized that combinations of TFs are required to reprogram T cell identity. This is motivated by the observation that most cell reprogramming strategies require multiple factors.
We therefore set out to identify TFs that collaborate within exhaustion-specific programs.
Fast forwarding through a lot of work: we screened ~3,500 TFs, profiled ~750K cells with perturb-SHARE-seq, and identified KLF2 as a potent context-specific repressor of exhaustion programs.
These same programs are associated with favorable outcomes in CAR T therapy, motivating us to dig deeper.
We then engineered this interaction: we synthetically tethered RUNX2 and KLF2, forcing their collaboration to create a stronger and more specific gene expression response.
@biorxivpreprint.bsky.social RUNX is a "master collaborator" in T cells, where RUNX2:KLF2 interactions specify exhaustion, which can be perturbed by mutating the RUNX2:KLF2 interface or amp'ed by synthetic tethering of RUNX2 to KLF2 @jbuenrostro.bsky.social @harvard.edu
doi.org/10.64898/202...
We next asked: how does KLF2 work?
Using perturb-SHARE-seq, we applied seq2PRINT and found that RUNX TFs are highly collaborative, enabling cells to access diverse regulatory programs. Additional experiments revealed that RUNX2 and KLF2 physically interact to regulate key exhaustion genes. 🧬
Can we control cells by forcing TFs to collaborate?
Interested? Read our preprint! An awesome collaboration with
@cgersbach.bsky.social, @igvfconsortium.bsky.social, Christian McRoberts Amador, @rachel-savage.bsky.social:
www.biorxiv.org/content/10.6...
🧵
A little late to Bluesky but my postdoc work w/ @jbuenrostro.bsky.social now out in @nature.com
"Epigenetic memory of colitis promotes tumour growth"
www.nature.com/articles/s41...
We wanted to understand how transient inflammation can create an increase in cancer risk, even after full recovery 🧵