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 1

Validation of the LRRK2-N2081D KI mouse model of CD.

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Validation of the LRRK2-N2081D KI mouse model of CD. (A) Odds ratios (OR...
(A) Odds ratios (ORs) for the associations of the G2019S and N2081D LRRK2 variants with CD and PD. (B) Schematic representation of the LRRK2 protein, highlighting the protein domains and locations of disease-associated variants. (C and E) Normalized percentage of body weight during acute DSS treatment in male and female mice (male mice: n = 22 WT, n = 15 G2019S, n = 24 N2081D; female mice: n = 17 each WT and G2019S, n = 19 N2081D). * indicates a significant difference between WT and either G2019S or N2081D; + indicates a significant difference between N2081D and G2019S. (D and F) Survival curves of male and female mice. Animals were sacrificed when humane endpoints were surpassed. Male mice showed significantly different survival among genotypes (****P < 0.0001); female mice showed no significant differences. (G and J) Measurements of colon length and representative images of male and female mice, respectively. (H and K) Stool lipocalin-2 measured by ELISA in male and female mice (male mice: n = 10 WT, n = 9 G2019S, n = 8 N2081D; female mice: n = 10 each WT and G2019S, n = 8 N2081D). (I and L) Spleen weight recorded at sacrifice for male and female mice (male mice: n = 21 WT, n = 15 G2019S, n = 24 N2081D; female mice: n = 16 WT, n = 17 G2019S, n = 19 N2081D). (M and N) Representative images and histological scoring of intestinal inflammation and damage in male and female mice (male mice: n = 13 WT, n = 9 each G2019S and N2081D; female mice: n = 12 WT, n = 10 each G2019S and N2081D). One-way ANOVA with Tukey’s post hoc test was used for C, E, F, and H–N; log-rank (Mantel-Cox) test was used for survival curves in D and F. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. EUR, European; meta, meta-analysis; UKBB, UK Biobank.