novel target space
Our platform identifies small molecules that selectively modulate the mRNA translation of target proteins. The novel target space enables the discovery of drugs that can either decrease or increase the production of proteins in a tissue selective manner.
Multiple specific mechanisms regulating mRNA translation
Cells have specific mechanisms that regulate the translation of individual mRNAs. We target proteins that have specific biological roles in coordinating and regulating translation in a selective way, allowing for the discovery of small molecules that modulate the translation of a target protein.
Monitoring mRNA translation
Our technology platform enables us to monitor mRNA translation of specific proteins or a group of proteins inside cells, visualizing when, where and how much of a target protein is being made in a disease model, in real time.
Translation of Collagen I mRNA in Human lung fibroblasts
- A and B. Monitoring of Collagen I mRNA translation in human lung fibroblasts using a pair of tRNA which specifically decodes Collagen I mRNA. (A) Low level of translation of Collagen I in resting, healthy fibroblasts; (B) High levels of Collagen I protein synthesis in activated, disease-model of lung fibrosis.
- C and D. Accumulation of Collagen I protein after five days. (C) Low levels of Collagen I protein accumulated in healthy lung fibroblasts; (D) High amount of Collagen I protein accumulated in cells in a disease model of lung fibrosis.
mRNA Translation and Target ID technologies
We have gained tremendous expertise, allowing us to build technologies around mRNA translation control. Our screening platform centers on a novel target space, including mRNA processing, transport, stability, localization and protein translation regulation. Compounds act on proteins and small RNAs, which regulate RNA rate and protein translation.
Target ID TechnologiesTranslation ControlQuality assurance of labeled tRNAAnalysis of interactions inside cells of mRNA and the proteins that bind to itIdentification of the compound's effect on mRNA and tRNA modificationsAnalysis of the half-life of target mRNA and protein to compoundIdentification of proteins which are bound by compoundSpecific isolation of mRNA and its associated proteins in response to compoundRNA sequencing of untreated versus compound treated cellsA score for compounds' effect on global translationRibosome occupancy on mRNAProprietary algorithm selects for gene-specific codon pairFluorescent probes detect single mRNAs inside cells
Chemically diverse hits target multiple regulatory mechanisms
We are discovering compounds that target mRNA maturation, steady state levels, localization and translation. Unlike other approaches that target the mRNA molecules or mRNA modifiers, our technology provides multiple shots on goal without depending on a single assumption about disease biology.
Hit compounds reduce Collagen I protein levels (upper panel, green), without affecting Collagen I mRNA levels (lower panel, red). Dots in the nucleus are COL1A1 transcription sites. Signal in the cytoplasm, is cytoplasmic mRNA. As a control, a compound which is known to inhibit transcription, reduces both protein and mRNA signals (ActD: Actinomycin D).
Detecting translation of a specific mRNA
Once the specific protein tRNA pair has been selected, we validate the assay by using siRNAs. We have been able to demonstrate Assay Specificity in all of our programs.
To ensure the FRET signal (light) is coming from Collagen I or c-Myc translation, we transfected cells with siRNA specific to the target mRNA. Most of the PSM signal in the siRNA transfected cells disappeared, emphasizing that the PSM signal has been mainly generated by Collagen I translation.
Tissue selectivity: a novel paradigm
Our small molecule drugs are tissue-specific; this stems from the fact that different cells regulate the production of the same protein in different ways.
Collagen I translation modulators regulate Collagen I translation in a tissue specific manner. A representative hit compound, compound X, inhibits Collagen I protein accumulation in the cell line used for the screen, WI-38 lung fibroblasts (panel A) and in primary human lung fibroblasts (panel B). However, the hit compound does not inhibit Collagen I translation in human primary skin fibroblasts (panel C).
We are able to identify compounds that are tissue specific in the regulation of Collagen I. We tested the effect of the compounds in liver fibroblasts (panel A) and in lung fibroblasts (panel B). Compounds A, B and C reduce the Collagen I in lung fibroblasts (panel B); compounds B and C do not affect Collagen I in liver fibroblasts, however compound A does show some inhibition of Collagen I in liver fibroblasts (panel A).
Safer drugs by design
The mechanisms regulating mRNA translation are mostly tissue selective. This enables us to discover drugs that work only in the specific tissue where there is disease. Anima's approach has been proven to discover compounds that are tissue selective and uses multiple secondary assays to confirm compounds' selectivity.
Our screening platform discovers a set of compounds that can affect only c-Myc protein and no other Myc family members as shown by Immunofluorescence in IF in A549 NSCLC (lung cancer cell line).