# Probing RAS-mediated signaling mechanisms with monobody inhibitors

> **NIH NIH R01** · MEDICAL UNIVERSITY OF SOUTH CAROLINA · 2020 · $551,022

## Abstract

Project Summary:
Cancer is a leading cause of death in the United States and worldwide. As such the President of the United
States has recently established the White House Cancer Moonshot Task Force, the mission of which is to
eliminate cancer as we know it. Part of this mission is to encourage development of novel cancer treatments.
This innovative multi-PI proposal represents a novel approach to this challenge. Oncogenic activation of the
RAS family of GTPases occurs in ~30% of cancers making it the most frequently mutated oncogene in human
cancers. Despite a great deal of progress in our understanding of the biochemistry of RAS and it's role in
tumorigenesis, development of effective therapeutic inhibitors of RAS to date has been disappointing. Thus,
there remains a critical need to develop targeted inhibitors of this oncoprotein for treatment of patients with
Ras-positive tumors. Using an unbiased, protein engineering approach, we have developed a highly specific
and potent inhibitor of H-RAS and K-RAS based on the monobody platform. Monobodies are single-domain
proteins of ~95 amino acids that achieve levels of affinity and selectivity similar to antibodies yet are insensitive
to the redox potential of their environment. High affinity monobodies have been isolated to a diverse array of
targets including the extracellular domain of receptors, kinases, steroid hormone receptors, and modular
protein domains. Using this protein-based monobody inhibitor as a powerful experimental tool, we will probe
the function of RAS in the tumorigenic process in ways that have not previously been possible. We propose
three major aims to accomplish our goal of interrogating RAS function in oncogenesis. In Aim 1, we will use
genetically encoded versions of our monobody inhibitor, termed NS1, to address unanswered questions
regarding RAS function. In particular, NS1 blocks RAS through binding an allosteric interface important for
dimerization of RAS and stimulation of signaling and transformation. Using NS1, we will address the
importance of RAS dimerization in activation of multiple RAS effector pathways as well as probe the isoform
specific difference in effector engagement. Aim 2 will employ a unique chemical-genetic approach to regulate
NS1 expression in vivo to address whether targeting this novel allosteric interface interferes with RAS-
dependent tumorigenesis. Building on our recent success with NS1, Aim 3 will develop isoform specific
inhibitory monobodies to each RAS isoform and determine their mechanism of action. These studies represent
a unique and powerful approach toward studying RAS and defining potential novel approaches to blocking
RAS action. Thus, our work has the potential to make a major impact on cancer therapy. In addition, this
project is highly relevant to the mission of National Cancer Institute's RAS Initiative at the Frederick National
Laboratory for Cancer Research which is charged with targeting RAS-dependent cancers as well as Pres...

## Key facts

- **NIH application ID:** 9977135
- **Project number:** 5R01CA212608-05
- **Recipient organization:** MEDICAL UNIVERSITY OF SOUTH CAROLINA
- **Principal Investigator:** SHOHEI KOIDE
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $551,022
- **Award type:** 5
- **Project period:** 2018-02-01 → 2022-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9977135

## Citation

> US National Institutes of Health, RePORTER application 9977135, Probing RAS-mediated signaling mechanisms with monobody inhibitors (5R01CA212608-05). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9977135. Licensed CC0.

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