2. Academic Validation
  3. A Hotspot Phosphorylation Site on SHP2 Drives Oncoprotein Activation and Drug Resistance

A Hotspot Phosphorylation Site on SHP2 Drives Oncoprotein Activation and Drug Resistance

  • bioRxiv. 2025 Jun 16:2025.06.11.659120. doi: 10.1101/2025.06.11.659120.
Prashath Karunaraj 1 2 3,  Remkes Scheele 1,  Malcolm L Wells 4,  Ruchita Rathod 1 5,  Sophia Abrahamson 1,  Lila C Taylor 1 2 3,  Ipek S Gokulu 1 2 3,  Lamia Chowdhury 1,  Abiha Kazmi 1 6,  Weixiao Song 7,  Peter Hornbeck 8,  Jing Li 8,  Anum Glasgow 4,  Neil Vasan 1 9 2 3
Affiliations

Affiliations

  • 1 Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, NY, USA.
  • 2 Department of Medicine, New York University Grossman School of Medicine, New York, NY, USA.
  • 3 Perlmutter Cancer Center, NYU Langone Health, New York, NY, USA.
  • 4 Department of Biochemistry and Molecular Biophysics, Columbia University Irving Medical Center, New York, NY, USA.
  • 5 Memorial Sloan Kettering Cancer Center, New York, NY, USA.
  • 6 SUNY Downstate Health Sciences University, Brooklyn, NY, USA.
  • 7 WuXi AppTec, Suzhou, China.
  • 8 Cell Signaling Technology, Danvers, MA, USA.
  • 9 Department of Medicine, Division of Hematology/Oncology, Columbia University Irving Medical Center, New York, NY, USA.
PMID: 40667115 DOI: 10.1101/2025.06.11.659120
Abstract

SHP2 is a Phosphatase and a critical mediator of receptor tyrosine kinase (RTK)-driven Ras/mitogen-activated protein kinase (MAPK) signaling. Despite promising preclinical data, SHP2 inhibitors have shown minimal clinical efficacy, with no defined clinical mechanisms of primary resistance. Here, we elucidate phosphorylation of SHP2 at tyrosine 62 (pY62) as a hotspot phosphorylation site in the proteome and RTK-driven tumor types in patients. We demonstrate that Src family kinases directly phosphorylate SHP2 at Y62, downstream of but not directly phosphorylated by RTKs. Using biochemical and biophysical analyses, we show that SHP2 Y62D enforces an open, active conformation, resulting in constitutive Phosphatase activation that is sufficient to activate MAPK signaling and confer resistance to allosteric SHP2 inhibitors. These findings establish that SHP2 pY62 is a phosphorylation hotspot phenocopying mutational activation, a mechanism of primary resistance to SHP2 inhibitors, and a Cancer drug target distinct from wildtype SHP2.

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