Fig. 6. LNA043 penetrates damaged cartilage and reverts the OA transcriptional signature in FIH study patients.
a, Representative images of LNA043 detection by IHC in TKR cartilage from patients with OA in the FIH trial 2 h post-LNA043 injection (left) or after placebo treatment (right). LNA043 (brown stain) showed deeper penetration into damaged (red bracket) than intact cartilage (green bracket). Cartilage from placebo-injected patients showed no LNA043-specific staining. Unspecific staining was seen in all specimens at the tidemark (arrows). The arrowheads indicate the superficial tear characteristic of damaged cartilage. These images are representative of 21 (LNA043 injection) and 49 (placebo treatment) IHC-stained cartilage samples. Scale bar, 600 μm. b, Transcriptomics study design. Damaged and undamaged articular cartilage samples were profiled from patients with OA injected with placebo (n = 7) or 20 mg LNA043 at 2 h, 7 d or 21 d (n = 3 each) before TKR. c, Left, heatmap showing the expression changes expressed as log2[fold change] (log2[FC]) of the 28 genes statistically significantly regulated by LNA043. OA damage refers to damaged versus undamaged cartilage in the placebo group; 2 h, 7 d and 21 d refer to damaged cartilage with LNA043 compared with placebo treatment. Numbers below the heatmap indicate the number of significantly regulated genes for each timepoint comparison. The 23 genes at the top are upregulated by LNA043, and the 5 genes at the bottom are downregulated by LNA043. Right, temporal expression profiles of three genes significantly and consistently downregulated by LNA043, expressed as log2[counts per million reads] (log2[CPM]). Statistical significance was determined by limma moderated t-test (defined as a Benjamini–Hochberg adjusted P value of ≤0.05 versus placebo; *PFN1 = 8.40 × 10−5 (2 h); *PFN11.57 × 10−9 (21 d); *PSPP1/OPN = 1.09 × 10−4; *PDNER = 0.012145). The data are presented as means ± s.d. d, Significant concordance between curated public OA disease signatures and FIH-derived OA signatures. LNA043 gene signatures inversely correlate with OA disease signatures. These were generated from the SkeletalVis repository and from published transcriptomics studies. Enrichment is expressed as the negative log10 of the false discovery rate [FDR] from GSEA. The signs indicate the direction of enrichment. Blue color intensity indicates enrichment in downregulated genes, and red color enrichment in upregulated genes. EGFR1, epidermal growth factor receptor 1. e, Reversal of OA-associated transcriptome changes in the FIH study by LNA043. Left, heatmaps of expression changes (log2[FC]) for genes regulated with a nominal limma moderated t-test P ≤ 0.05. Below the heatmaps are boxplots summarizing the log2[FC] for all genes in the heatmap per group. Central lines and box edges represent median values and upper and lower quartiles, respectively, and the whiskers represent 1.5× the interquartile range. The numbers of genes counter-regulated by LNA043 are shown in the bar plots below the boxplots. Right, temporal transcriptional response to LNA043 for OA-relevant pathway genes (expressed as the mean log2[CPM] ± s.d.). Statistical significance was defined as in c.