ANKRD46

ANKRD46 is a direct target of multiple miRNAs—miR-21, miR-451, and miR-25-3p—each of which binds its 3′ UTR to repress protein expression, as validated by luciferase reporter assays and Western blot across breast cancer, hepatocellular carcinoma, and murine luminal epithelial systems (PMID:21219636, PMID:25542822, PMID:37485755). Derepression of ANKRD46, whether by anti-miR-21 treatment or circular RNA sponging of miR-25-3p, suppresses cancer cell proliferation, migration, and invasion, and inhibits tumor growth in xenograft models, consistent with a tumor-suppressive function (PMID:25550434, PMID:37485755). In murine uterine luminal epithelium, miR-451–mediated repression of Ankrd46 is required for normal embryo implantation (PMID:25542822).

1. 2011 Medium

   Establishing ANKRD46 as a miRNA target: the discovery that miR-21 directly binds the ANKRD46 3′ UTR and represses its protein expression provided the first functional annotation for this otherwise uncharacterized ankyrin-repeat protein.

   Evidence Luciferase reporter assay with 3′ UTR construct and Western blot after LNA-mediated miR-21 knockdown in breast cancer cell lines

   PMID:21219636

   Open questions at the time

   • No seed-site mutagenesis was performed to confirm the precise miR-21 binding element
   • The intrinsic molecular activity of ANKRD46 protein remained unknown
   • No direct phenotypic consequence of ANKRD46 manipulation (independent of miR-21) was tested
2. 2014 Medium

   Extending miRNA regulation to a physiological context: showing that miR-451 targets Ankrd46 in murine luminal epithelium and that miR-451 loss-of-function impairs embryo implantation placed ANKRD46 in a developmental pathway beyond cancer.

   Evidence Dual-luciferase reporter assay plus miR-451 sponge/inhibitor studies in vitro and in vivo mouse implantation model

   PMID:25542822

   Open questions at the time

   • Whether Ankrd46 overexpression or knockdown alone alters implantation was not directly tested
   • The downstream effectors of Ankrd46 in luminal epithelium are unknown
3. 2014 Medium

   Functional reinforcement: demonstrating that anti-miR-21 free uptake induces ANKRD46 expression and inhibits hepatocellular carcinoma cell growth corroborated ANKRD46 as a functionally relevant miR-21 target with growth-suppressive properties in a second cancer type.

   Evidence Anti-miR-21 free uptake assay with ANKRD46 induction and cell growth inhibition in HCC cells

   PMID:25550434

   Open questions at the time

   • Growth suppression was not tested by ANKRD46 overexpression alone, so the effect could be partly miR-21-independent
   • Single cell line without rescue experiment
4. 2023 Medium

   Defining a ceRNA axis: the circ_PRDM5/miR-25-3p/ANKRD46 circuit demonstrated that ANKRD46 derepression suppresses breast cancer proliferation, migration, and invasion in vitro and tumor growth in vivo, strengthening its candidacy as a tumor suppressor.

   Evidence Dual-luciferase assay, Western blot, miRNA sponge overexpression/knockdown, and xenograft tumor assay in breast cancer models

   PMID:37485755

   Open questions at the time

   • ANKRD46 knockout/knockdown alone was not used to confirm necessity for the tumor-suppressive phenotype
   • The protein-level mechanism by which ANKRD46 suppresses proliferation and migration is unknown
   • No structural or interactome data exist to explain ANKRD46 function at the molecular level
5. 2025 Low

   Pathway association: linking ANKRD46 expression to the IL6-JAK-STAT3 signaling pathway in keloid and type 2 diabetes contexts suggested a broader role in fibrotic and metabolic disease, though without direct mechanistic dissection.

   Evidence Western blot, qRT-PCR, IHC, and flow cytometry in mouse models; pathway identification via machine learning (LASSO/SVM-RFE)

   PMID:41346282

   Open questions at the time

   • No direct evidence that ANKRD46 modulates IL6-JAK-STAT3 signaling (association only, not causation)
   • Computational biomarker discovery without mechanistic validation
   • Single study, not independently replicated

• The intrinsic molecular activity of ANKRD46—its binding partners, enzymatic or scaffolding function, and the mechanism by which it suppresses cell proliferation and migration—remains entirely uncharacterized.
• No direct interactome, structural, or biochemical characterization of ANKRD46 protein exists
• Whether ANKRD46 acts as a scaffold, adaptor, or transcriptional regulator is unknown
• The pathway(s) through which ANKRD46 exerts tumor-suppressive effects have not been identified