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Updated: Dec 13, 2025
Assessing Cellular Target Engagement by SHP2 PTPN11 Phosphatase Inhibitors
Published on: July 17, 2020
Assessing Cellular Target Engagement by SHP2 (PTPN11) Phosphatase Inhibitors
Lester J Lambert1, Celeste Romero1, Douglas J Sheffler1
1Cancer Metabolism & Signaling Networks Program, NCI-Designated Cancer Center, Sanford Burnham Prebys Medical Discovery Institute.
Abstract:
The Src-homology 2 (SH2) domain-containing phosphatase 2 (SHP2), encoded by the PTPN11 proto-oncogene, is a key mediator of receptor tyrosine kinase (RTK)-driven cell signaling, promoting cell survival and proliferation. In addition, SHP2 is recruited by immune check point receptors to inhibit B and T cell activation. Aberrant SHP2 function has been implicated in the development, progression, and metastasis of many cancers. Indeed, small molecule SHP2 inhibitors have recently entered clinical trials for the treatment of solid tumors with Ras/Raf/ERK pathway activation, including tumors with some oncogenic Ras mutations. However, the current class of SHP2 inhibitors is not effective against the SHP2 oncogenic variants that occur frequently in leukemias, and the development of specific small molecules that target oncogenic SHP2 is the subject of current research. A common problem with most drug discovery campaigns involving cytosolic proteins like SHP2 is that the primary assays that drive chemical discovery are often in vitro assays that do not report the cellular target engagement of candidate compounds. To provide a platform for measuring cellular target engagement, we developed both wild-type and mutant SHP2 cellular thermal shift assays. These assays reliably detect target engagement of SHP2 inhibitors in cells. Here, we provide a comprehensive protocol of this assay, which provides a valuable tool for the assessment and characterization of SHP2 inhibitors.
Insights
We developed cellular thermal shift assays to measure target engagement for SHP2 inhibitors. This assay is crucial for developing new cancer therapies targeting SHP2 oncogenic variants.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- SHP2 (Src-homology 2 domain-containing phosphatase 2) is a key regulator of receptor tyrosine kinase signaling, impacting cell survival, proliferation, and immune responses.
- Aberrant SHP2 activity is linked to cancer development, progression, and metastasis.
- Existing SHP2 inhibitors show limited efficacy against leukemia-associated oncogenic SHP2 variants, necessitating novel therapeutic strategies.
Purpose of the Study:
- To develop a cellular assay for measuring target engagement of SHP2 inhibitors.
- To provide a tool for assessing SHP2 inhibitors against both wild-type and mutant SHP2 forms.
- To facilitate the discovery of new SHP2-targeted cancer therapies.
Main Methods:
- Development of wild-type and mutant SHP2 cellular thermal shift assays (CETSA).
- Utilized CETSA to detect cellular target engagement of SHP2 inhibitors.
- Validated the assay's reliability in measuring inhibitor binding to SHP2 within cells.
Main Results:
- Established robust cellular thermal shift assays for SHP2.
- Demonstrated the ability of these assays to reliably detect target engagement of SHP2 inhibitors in cellular contexts.
- Provided a comprehensive protocol for the SHP2 CETSA.
Conclusions:
- The developed SHP2 cellular thermal shift assays are valuable tools for drug discovery and characterization.
- These assays enable the assessment of SHP2 inhibitor efficacy at the cellular level, particularly for oncogenic variants.
- This platform supports the development of novel SHP2-targeted cancer therapeutics.
