Figure 5. Sik targeting drives a transcriptional state resembling Lkb1 loss in mouse models and human lung tumors.

A. Hierarchical clustering of tumors derived from KT and KT;Lkb1^flox/flox mice as well as KT;H11^LSL-Cas9 mice transduced with lentiviral sgRNA vectors targeting either Sik1 alone or Sik1–3.

B. Heatmap of differentially expressed genes in KT;Lkb1^flox/flox tumors relative to KT tumors depicting the conservation of differential gene expression in Sik-targeted tumors. Significant differential expression defined as an absolute log2(fold change) > 1 and q < 0.05.

C,D. Venn diagrams depicting conservation of differential gene expression relative to KT tumors between KT;Lkb1^flox/flox tumors and Sik-targeted tumors initiated in KT;H11^LSL-Cas9 mice. Significance cutoffs and hypergeometric test p-values are indicated.

E. Direct comparison of pathways that are higher in KT;Lkb1^flox/flox tumors and Sik-targeted tumors relative to KT tumors. Each dot corresponds to a gene set derived from the mSigDB Hallmarks module. NES signifies the normalized enrichment score calculated for each gene set using GSEA. Pathways that are significantly enriched in both tumor genotypes are colored red.

F. GSEA with genes that are higher in Sik-deficient tumors on the comparison of K;Lkb1^flox/flox and K tumors (7).

G. CDF plot of the distributions of signature scores for human tumors stratified by LKB1 genotype using only the downregulated subset of genes within the Sik deficiency signature. Cohort size and Kolmogorov-Smirnov test p-value are indicated (20). Background shading delineates the Sik signature score tertiles.

H. Stacked barplot summarizing the distribution of LKB1 mutant human tumors across tertiles of Sik signature scores. Chi-square test p-value is indicated.

I. GSEA using subsets of upregulated or downregulated genes within the Sik signature genes performed on external gene expression data derived from the re-expression of LKB1 in LKB1-deficient cell lines (A549, H2122, H2126) or the knocking down of LKB1 expression in an LKB1-proficient cell line (H1650)(7,14,56).