Targeting specific kinase substrates rescues increased colitis severity induced by the Crohn’s disease–linked LRRK2-N2081D variant
George R. Heaton, … , Inga Peter, Zhenyu Yue
George R. Heaton, … , Inga Peter, Zhenyu Yue
Published October 1, 2025
Citation Information: J Clin Invest. 2025;135(19):e190017. https://doi.org/10.1172/JCI190017.
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Research Article Gastroenterology Genetics

Targeting specific kinase substrates rescues increased colitis severity induced by the Crohn’s disease–linked LRRK2-N2081D variant

Abstract

LRRK2 contains a kinase domain where the N2081D Crohn’s disease (CD) risk and the G2019S Parkinson’s disease (PD) pathogenic variants are located. It is not clear how the N2081D variant increases CD risk or how these adjacent mutations give rise to distinct disorders. To investigate the pathophysiology of the CD-linked LRRK2 N2081D variant, we generated a knock-in (KI) mouse model and compared its effects with those of the LRRK2-G2019S mutation. Lrrk2N2081D KI mice demonstrated heightened sensitivity to induced colitis, resulting in more severe intestinal damage than in Lrrk2G2019S KI and WT mice. Analysis of colon tissue revealed distinct mutation-dependent LRRK2 RAB substrate phosphorylation, with significantly elevated phosphorylated RAB10 levels in Lrrk2N2081D mice. In cells, we demonstrated that the N2081D mutation activates LRRK2 through a mechanism distinct from that of LRRK2-G2019S. We also found that proinflammatory stimulation enhances LRRK2 kinase activity, leading to mutation-dependent differences in RAB phosphorylation and inflammatory responses in dendritic cells (DCs). Finally, we show that knockout of Rab12, but not pharmacological LRRK2 kinase inhibition, significantly reduced colitis severity in Lrrk2N2081D mice. Our study characterizes the pathogenic mechanisms of LRRK2-linked CD, highlights structural and functional differences between disease-associated LRRK2 variants, and suggests RAB proteins as promising therapeutic targets for modulating LRRK2 activity in CD treatment.

Authors

George R. Heaton, Xingjian Li, Xianting Li, Xiaoting Zhou, Yuanxi Zhang, Duc Tung Vu, Marc Oeller, Ozge Karayel, Quyen Q. Hoang, Meltem Ece Kars, Nitika Kamath, Minghui Wang, Leonid Tarassishin, Matthias Mann, Inga Peter, Zhenyu Yue

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Figure 2

Kinase activity characterization of PD- and CD-linked LRRK2 variants.

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Kinase activity characterization of PD- and CD-linked LRRK2 variants. (A...
(A) Representative IBs of colon and brain tissue lysates from WT, G2019S, and N2081D KI mice. (B) Quantification of endogenous LRRK2 S1292 autophosphorylation and Rab substrate phosphorylation. (C) Representative IBs of HEK293 cells expressing LRRK2 variants. (D) Quantification of LRRK2 S1292 autophosphorylation and phosphorylation of endogenous Rab substrates in HEK293 cells (n = 5 independent experiments). (E) Structural modeling of LRRK2 domains (LRR, ROC, COR, and kinase) based on the full-length structure, with the positions of N2081 and G2019 residues highlighted. (F) Representative IBs of HEK293 cells expressing LRRK2 mutants at N2081 and N1269 residues. (G) Quantification of LRRK2 autophosphorylation and Rab phosphorylation (n = 5 independent experiments). One-way ANOVA with Tukey’s post hoc test was used for B; 1-way ANOVA with Dunnett’s post hoc test was used for D and G. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.