The Minor Allele Of A Single Nucleotide Polymorphism In Sh2b3 Promotes Hypertension And Renal Dysfunction In Mice.

SH2B3, also known as LNK, is an adaptor protein that negatively regulates growth factor and cytokine signaling. The minor allele of a single nucleotide polymorphism in SH2B3 (rs3184504) encodes a tryptophan (Trp) at amino acid 262 as opposed to arginine (Arg) and is strongly associated with hypertension in genome-wide association studies. Whether this variant is causal and how it impacts hypertension development and end-organ damage is unknown. We used CRISPR-Cas9 to engineer mice homozygous for the major and minor alleles of this SH2B3 polymorphism, resulting in Arg/Arg and Trp/Trp mice, respectively. Trp/Trp mice exhibited increased systolic blood pressure (SBP) by radiotelemetry during weeks 3-4 of angiotensin II (Ang II) infusion compared to Arg/Arg mice (nighttime SBP 168 vs 158 mm Hg, respectively). Renal dysfunction was also exacerbated in Ang II-treated Trp/Trp compared to Arg/Arg mice, as evidenced by significantly increased urinary albumin/creatinine ratio (0.41 vs 0.17), renal perivascular fibrosis (fibrosis score 2.0 vs 1.0), and renal macrophages (8,341 vs 6,413 per kidney). In addition, renal CD8 + T cells from Ang II-treated Trp/Trp mice produced significantly more IFNγ compared to Arg/Arg controls (median fluorescence intensity 20,455 vs 14,134), and ex vivo stimulated splenic CD8 + T cells from Trp/Trp compared to Arg/Arg mice made 2.7-fold more IFNγ. To determine a mechanism for increased T cell IFNγ production, dendritic cells and naïve T cells from Trp/Trp and Arg/Arg mice were co-cultured in different combinations. The greatest increase in GM-CSF-induced IFNγ production occurred when both dendritic cells and T cells came from Trp/Trp mice (3.4-fold greater than both cell types from Arg/Arg mice). This effect appears to be due to loss of SH2B3-mediated suppression of GM-CSF signaling, as overexpression of Trp-encoding SH2B3 in HEK cells exhibited significantly less repression of GM-CSF-induced Stat5 activation compared to Arg-encoding SH2B3. Taken together, these findings suggest that the Trp encoding allele of rs3184504 is a causal variant promoting blood pressure elevation and renal dysfunction, at least in part through loss of SH2B3-mediated repression of T cell IFNγ production.