Fig 2: CCL21 promotes GC cell growth in vitro. (A, B) CCR7 and CLDN11 genes were significantly upregulated in GC-CCL21 cells. (C) CCL21 significantly promoted the proliferation of GC cells. (D, E) CCL2 have no significant effect on apoptosis of GC cells.

Fig 3: Immune infiltration and IHC. (A, B) immune infiltration for all samples. (C) CCL21 and the relative abundance of immune cells. (D) CCL21 and CLDN11 were significantly higher in the metastatic lesion tissue than in the primary lesion tissue. (E, F) Compared to LN group, CCL21 and CLDN11 were significantly upregulated in MTT group.

Fig 4: Dysregulated striatal oligodendrocyte gene expression is reversed in response to mHTT lowering in the brain(A) Volcano plot showing DEGs in the striatum of PBS-treated YAC128 mice (n = 6) compared to WT mice (n = 6) at the 6-month interval (12 months of age). The fold change of DEGs (log2 transformed) are plotted versus adjusted p values (−log10 transformed). DEGs significantly downregulated compared to WT are shown in blue, while those significantly upregulated are shown in red (FDR <0.1). The total number of DEGs that are downregulated (left) or upregulated (right) is indicated. The top 10 most significantly down- and upregulated DEGs are annotated. Msantd1 was the most significantly upregulated gene (−log10(adjusted p value) = 136.77) and exhibited the greatest fold change (log2(fold change) = 3.587); however, it was excluded from the plot as it fell outside the range of most other genes. Horizontal dotted line indicates the significance threshold. The top 100 downregulated and upregulated striatal DEGs for this comparison are available in Table S1. (B) Balloon plot depicting selected GO biological process terms significantly enriched for DEGs that are downregulated (blue) or upregulated (red) (FDR <0.1) in PBS-treated YAC128 mice compared to WT mice at the 6-month interval. The x axis represents the –log10(p value) of enrichment after multiple testing correction. Balloon color reflects the degree of overlap between DEGs and the corresponding GO term, while balloon size represents the total number of genes associated with the GO term. (C) Scatterplot (left) illustrating the transcriptome-wide association of Z-statistics between YAC128 (PBS) vs. WT (naive) and YAC128 (PBS) vs. YAC128 (HTT ASO) comparisons. Pearson’s correlation analysis was performed to evaluate the strength and direction of relationships between variables. The reversal correlation coefficient (r) and corresponding nominal p value are shown. Dashed gray lines indicate approximate FDR = 0.1 thresholds, while the regression line is shown in black. Genes fully rescued by HTT ASO treatment are highlighted in green, partially rescued in blue, exacerbated in orange, and minimally changed in gray. The top five fully rescued upregulated and downregulated genes are annotated. Vertical bar plot (right) depicting the number of genes from the HD signature (PBS-treated YAC128 vs. WT); 1,127 genes from (A) categorized based on their response to HTT ASO treatment at the 6-month interval. Genes are classified as fully rescued (green), partially rescued (blue), exacerbated (orange), or minimal change (gray). (D) Balloon plot showing selected terms from the HDSigDB gene sets significantly enriched for DEGs that are upregulated in HTT-ASO-treated compared to PBS-treated YAC128 mice. The top 100 downregulated and upregulated striatal DEGs for this comparison is available in Table S2. The x axis represents the −log10(p value) of enrichment after Benjamini-Hochberg correction. Balloon color indicates the odds ratio, while balloon size corresponds to the combined score. (E) Hierarchical clustering of cell types and bullseye plot outputs from CSEA, illustrating the overlap of transcripts enriched and/or specific to certain cell types with nominally significant DEGs (p < 0.05) upregulated in HTT-ASO-treated compared to PBS-treated YAC128 mice. Hexagon colors indicate Benjamini-Hochberg corrected p values, ranging from yellow (adjusted p = 0.1) to burgundy (adjusted p = 0). (F) Heatmaps displaying Z-statistics for the top 20 DEGs enriched in MSNs (left) and oligodendrocytes (right) across groups. Color scale ranges from red (positive Z-statistics) to blue (negative Z-statistics). (G, H) Representative immunoblots and relative quantification of (G) PLP1 (one-way ANOVA: p = 0.051; unpaired t test: p = 0.052 HTT ASO vs. PBS) and (H) CLDN11 (one-way ANOVA: p = 0.066; unpaired t test: #p = 0.043 HTT ASO vs. PBS) levels in the striatum of naive WT mice (N = 6) or YAC128 mice treated with either PBS (N = 8) or HTT ASO (N = 8). PLP1 and CLDN11 levels were normalized to actin and are presented relative to levels measured in WT naive controls for each respective protein. Boxes show the IQR, whiskers extend from the minimum to maximum value, horizontal lines show the median, and crosses show the mean.

Fig 5: Dysregulated striatal oligodendrocyte gene expression is reversed in response to mHTT lowering in the brain(A) Volcano plot showing DEGs in the striatum of PBS-treated YAC128 mice (n = 6) compared to WT mice (n = 6) at the 6-month interval (12 months of age). The fold change of DEGs (log2 transformed) are plotted versus adjusted p values (−log10 transformed). DEGs significantly downregulated compared to WT are shown in blue, while those significantly upregulated are shown in red (FDR <0.1). The total number of DEGs that are downregulated (left) or upregulated (right) is indicated. The top 10 most significantly down- and upregulated DEGs are annotated. Msantd1 was the most significantly upregulated gene (−log10(adjusted p value) = 136.77) and exhibited the greatest fold change (log2(fold change) = 3.587); however, it was excluded from the plot as it fell outside the range of most other genes. Horizontal dotted line indicates the significance threshold. The top 100 downregulated and upregulated striatal DEGs for this comparison are available in Table S1. (B) Balloon plot depicting selected GO biological process terms significantly enriched for DEGs that are downregulated (blue) or upregulated (red) (FDR <0.1) in PBS-treated YAC128 mice compared to WT mice at the 6-month interval. The x axis represents the –log10(p value) of enrichment after multiple testing correction. Balloon color reflects the degree of overlap between DEGs and the corresponding GO term, while balloon size represents the total number of genes associated with the GO term. (C) Scatterplot (left) illustrating the transcriptome-wide association of Z-statistics between YAC128 (PBS) vs. WT (naive) and YAC128 (PBS) vs. YAC128 (HTT ASO) comparisons. Pearson’s correlation analysis was performed to evaluate the strength and direction of relationships between variables. The reversal correlation coefficient (r) and corresponding nominal p value are shown. Dashed gray lines indicate approximate FDR = 0.1 thresholds, while the regression line is shown in black. Genes fully rescued by HTT ASO treatment are highlighted in green, partially rescued in blue, exacerbated in orange, and minimally changed in gray. The top five fully rescued upregulated and downregulated genes are annotated. Vertical bar plot (right) depicting the number of genes from the HD signature (PBS-treated YAC128 vs. WT); 1,127 genes from (A) categorized based on their response to HTT ASO treatment at the 6-month interval. Genes are classified as fully rescued (green), partially rescued (blue), exacerbated (orange), or minimal change (gray). (D) Balloon plot showing selected terms from the HDSigDB gene sets significantly enriched for DEGs that are upregulated in HTT-ASO-treated compared to PBS-treated YAC128 mice. The top 100 downregulated and upregulated striatal DEGs for this comparison is available in Table S2. The x axis represents the −log10(p value) of enrichment after Benjamini-Hochberg correction. Balloon color indicates the odds ratio, while balloon size corresponds to the combined score. (E) Hierarchical clustering of cell types and bullseye plot outputs from CSEA, illustrating the overlap of transcripts enriched and/or specific to certain cell types with nominally significant DEGs (p < 0.05) upregulated in HTT-ASO-treated compared to PBS-treated YAC128 mice. Hexagon colors indicate Benjamini-Hochberg corrected p values, ranging from yellow (adjusted p = 0.1) to burgundy (adjusted p = 0). (F) Heatmaps displaying Z-statistics for the top 20 DEGs enriched in MSNs (left) and oligodendrocytes (right) across groups. Color scale ranges from red (positive Z-statistics) to blue (negative Z-statistics). (G, H) Representative immunoblots and relative quantification of (G) PLP1 (one-way ANOVA: p = 0.051; unpaired t test: p = 0.052 HTT ASO vs. PBS) and (H) CLDN11 (one-way ANOVA: p = 0.066; unpaired t test: #p = 0.043 HTT ASO vs. PBS) levels in the striatum of naive WT mice (N = 6) or YAC128 mice treated with either PBS (N = 8) or HTT ASO (N = 8). PLP1 and CLDN11 levels were normalized to actin and are presented relative to levels measured in WT naive controls for each respective protein. Boxes show the IQR, whiskers extend from the minimum to maximum value, horizontal lines show the median, and crosses show the mean.