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Energy and Macronutrient Metabolism (Poster Session)

(P06-012-24) Interaction of the Src Homology Phosphatase 2 (Shp2) and Vimentin Modulates Glomerular Podocyte Function

Sunday, June 30, 2024
12:45 PM – 1:45 PM CT
Location: Poster Board 260
Disclosure(s):
Grace LeBleu, RDN, MSc: No relevant financial relationship(s) with ineligible companies to disclose.


Objectives: The objective of this study is to uncover the molecular mechanism underlying hyperglycemia-induced podocyte dysfunction. Podocytes are insulin sensitive cells in the kidney and crucial regulators of kidney function. We investigated the interaction of Shp2 with vimentin and the implications on podocyte function. Vimentin is an intermediate filament protein involved in cell motility, and its assembly into polymer filaments is dictated by tyrosine phosphorylation.

Methods: We generated mice with podocyte-specific Shp2 disruption. Mice were fasted overnight, injected with saline or insulin, then sacrificed and kidney tissues were collected. Vimentin was immunoprecipitated (IP) from kidney lysates then immunoblotted (IB) for phosphor-tyrosine residues (pTyr). Additionally, we generated E11 podocyte cell lines with Shp2 knockdown and reconstituted them with the human wildtype isoform (KD-WT) or substrate trapping double mutant (KD-DM). Cells were stimulated with insulin followed by IP: vimentin and IB: pTyr. Moreover, we used a pharmacological inhibitor of Shp2 (SHP099) to evaluate vimentin phosphorylation and wound healing.

Results: Podocyte specific Shp2 knockout appears to increase insulin induced phosphorylation of vimentin, and Shp2 inactivation attenuates insulin-induced vimentin phosphorylation in differentiated podocytes. Moreover, insulin induces co-association of vimentin with Shp2 KD-DM and is abolished by treatment with vanadate, in keeping with an enzyme-substrate interaction. Furthermore, Shp2 pharmacological inhibition mitigates high glucose-induced motility of E11 podocytes.

Conclusions: We suggest Shp2 dephosphorylates vimentin in an insulin inducible manner. Shp2 may impart effects on cell motility by modulating vimentin phosphorylation.

Funding Sources: Research in the Haj laboratory was funded by the NIDDK (R01DK095359, R01DK090492), NIEHS (P42ES04699), NIFA (CA-D*-NTR-7836H), and the Nora Eccles Treadwell Foundation. Dr. Hsu was supported in part by the NIAAA (R21AA027633). Ms. LeBleu was supported in part by the Rucker Family Fellowship and the Sugar Association/Research DPG Pilot Grant Award.