Cunniff, Patrick J (February 2025) Mechanisms of Transcriptional Regulation by RUVBL1/2 and KLF5 in Pancreatic Ductal Adenocarcinoma. PhD thesis, Cold Spring Harbor Laboratory.
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Abstract
One essential attribute of the proto-oncogenic epigenetic reprogramming resulting from pancreatitis is the potent upregulation of KLF5. KLF5 is necessary for Kras-driven malignant transformation and is an acquired and sustained dependency in pancreatic ductal adenocarcinoma (PDAC). To better evaluate the role of KLF5 in PDAC, we implemented an acute degradation system in human cell line models of both classical and basal-like PDAC to profile nascent transcriptional effects following KLF5 perturbation. These efforts revealed that KLF5 occupies a subtype-specific cistrome, which allows it to differentially support lineage identity across subtypes, while uniformly driving proliferation. Furthermore, the subtype-specific KLF5 gene expression programs that we defined are specific to human malignant cells yet reflect patient-specific transcriptional signatures which contribute to PDAC heterogeneity. This suggests that dissection of the KLF5 regulatory circuit will yield insight into the mechanisms which govern PDAC identity and plasticity. To characterize the PDAC-relevant KLF5 transcriptional complex, we devised an integrated marker-based CRISPR screen-pulldown mass spectrometry assay. This assay revealed the ATPase RUVBL1/2 as a bona fide KLF5 coactivator. By performing systematic deletion mutations, we identified the RUVBL1/2 binding residues on KLF5. Additionally, small molecule inhibition of RUVBL1/2 disrupts the KLF5 interaction, and impairs KLF5 transcriptional activity. Interestingly, relative to KLF5-independent cells, KLF5-dependent PDAC cells exhibit an increased sensitivity to RUVBL1/2 inhibition. However, the RUVBL1/2 interaction is not specific to KLF5. RUVBL1/2 exhibits the capacity to bind several lineage transcription factors across diverse cancer contexts. Collectively, our data suggests that small molecule inhibition of the RUVBL1/2 complex is a mechanism for disrupting transcription factor interactions, and thus, lineage transcription factor activity.
| Item Type: | Thesis (PhD) |
|---|---|
| Subjects: | diseases & disorders > cancer diseases & disorders diseases & disorders > cancer > cancer types > pancreatic cancer diseases & disorders > cancer > cancer types |
| CSHL Authors: | |
| Communities: | CSHL labs > Vakoc lab School of Biological Sciences > Theses |
| Depositing User: | Kathleen McGuire |
| Date: | 11 February 2025 |
| Date Deposited: | 30 Mar 2026 19:34 |
| Last Modified: | 30 Mar 2026 19:34 |
| URI: | https://repository.cshl.edu/id/eprint/42130 |
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