Proteolytic pan-RAS Cleavage Leads to Tumor Regression in Patient-derived Pancreatic Cancer Xenografts
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Abstract
The lack of effective RAS inhibition represents a major unmet medical need in the treatment of pancreatic ductal adenocarcinoma (PDAC). Here, we investigate the anticancer activity of RRSP-DTB, an engineered biologic that cleaves the Switch I of all RAS isoforms, in KRAS-mutant PDAC cell lines and patient-derived xenografts (PDX). We first demonstrate that RRSP-DTB effectively engages RAS and impacts downstream ERK signaling in multiple KRAS-mutant PDAC cell lines inhibiting cell proliferation at picomolar concentrations. We next tested RRSP-DTB in immunodeficient mice bearing KRAS-mutant PDAC PDXs. Treatment with RRSP-DTB led to ≥95% tumor regression after 29 days. Residual tumors exhibited disrupted tissue architecture, increased fibrosis and fewer proliferating cells compared with controls. Intratumoral levels of phospho-ERK were also significantly lower, indicating in vivo target engagement. Importantly, tumors that started to regrow without RRSP-DTB shrank when treatment resumed, demonstrating resistance to RRSP-DTB had not developed. Tracking persistence of the toxin activity following intraperitoneal injection showed that RRSP-DTB is active in sera from immunocompetent mice for at least 1 hour, but absent after 16 hours, justifying use of daily dosing. Overall, we report that RRSP-DTB strongly regresses hard-to-treat KRAS-mutant PDX models of pancreatic cancer, warranting further development of this pan-RAS biologic for the management of RAS-addicted tumors.
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RRSP-DTB consists of a RAS/RAP1-specific endopeptidase from Vibrio vulnificus combined with the translocation B fragment of diphtheria toxin.
“RRSP–DTB is one such agent (Table 1), consisting of a RAS/RAP1-specific endopeptidase (RRSP) derived from the bacillus Vibrio vulnificus combined with translocation B fragment of diphtheria toxin (DTB)191.”
From The current state of the art and future trends in RAS-targeted cancer therapies (2022) · cites this paper as [191]
In immunocompetent mice, RRSP-DTB was not detectable in serum 16 hours after administration, suggesting that frequent dosing may be needed.
“Notably, when studied in immunocompetent mice, the toxin was not detectable in serum at 16 hours after administration, suggesting the need for frequent dosing191.”
From The current state of the art and future trends in RAS-targeted cancer therapies (2022) · cites this paper as [191]
Vidimar et al. showed that RRSP-DTB inhibits ERK signalling via RAS inactivation in KRAS-mutant PDAC models, producing at least 95% tumour regression without evidence of resistance.
“Vidimar et al.191 have shown that this agent inhibits ERK signalling via RAS inactivation in KRAS-mutant PDAC models, resulting in ≥95% tumour regression without evidence of resistance191.”
From The current state of the art and future trends in RAS-targeted cancer therapies (2022) · cites this paper as [191]
RRSP-DTB enters cells through HB-EGF-mediated endocytosis and cleaves RAS within the switch 1 region.
“This toxin enters cells via heparin-binding EGF-like growth factor (HB-EGF)-mediated endocytosis and subsequently cleaves RAS within the switch 1 region191.”
From The current state of the art and future trends in RAS-targeted cancer therapies (2022) · cites this paper as [191]
RRSP-DTB can cleave HRAS, NRAS and KRAS irrespective of whether they are GTP- or GDP-bound.
“Uniquely, RRSP–DTB is able to cleave HRAS, NRAS and KRAS irrespective of GTP or GDP binding191.”
From The current state of the art and future trends in RAS-targeted cancer therapies (2022) · cites this paper as [191]
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Cited by
Sorted by citations
The current state of the art and future trends in RAS-targeted cancer therapies
Punekar, S. R., Velcheti, V., Neel, B. G., & Wong, K.-K. (2022) · Nature Reviews Clinical Oncology · doi:10.1038/s41571-022-00671-9 · Cited by 549 · see citing works
- Authors
- Vidimar, V., Park, M., Stubbs, C. K., Ingram, N. K., Qiang, W., Zhang, S., Gursel, D., Melnyk, R. A., & Satchell, K. J. F.
- Affiliations
- Northwestern University (US)
- University of Toronto (CA)
- Corresponding author
- Satchell, Karla J.F.
- ORCID
- Venue
- Molecular Cancer Therapeutics
- Published
- 2022-02-28
- Volume
- 21
- Issue
- 5
- Pages
- 810-820
- ISSN
- 1535-7163 (ISSN-L)
- 1538-8514
- PMC
- PMC9933180
- OpenAlex
- W4220863628
- Cited by
- 10
- References
- 44
- Citations by year
- 2026: 3 · 2025: 4 · 2023: 2 · 2022: 1
- FWCI
- 0.73
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Molecular Cancer Therapeutics (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Chicago Biomedical Consortium Accelerator Award (A-013)
- NCI (P30CA060553)
- NCI Ruth L. Kirstenstein fellowship (T32 CA09560)
- Canadian Institutes of Health Research (366017)
- Topics
- Health Sciences › Medicine › Oncology › Pancreatic and Hepatic Oncology Research
- Life Sciences › Immunology and Microbiology › Immunology › Toxin Mechanisms and Immunotoxins
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Keywords
- pancreatic ductal adenocarcinoma, RAS inhibition, ERK signaling, patient-derived xenografts, tumor regression, target engagement, tumor fibrosis
- MeSH terms
- Animals, Biological Products, Disease Models, Animal, Humans, Mutation, Pancreatic Neoplasms, Proto-Oncogene Proteins p21(ras), Carcinoma, Pancreatic Ductal, Cell Line, Tumor, Mice, Heterografts
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