G4 Oligonucleotide-Based Chaperones of Heterogeneous Nuclear Ribonucleoprotein A1

TL;DR

Scientists designed new drug-like molecules that bind to a protein called hnRNP A1 and prevent it from forming harmful clumps linked to neurodegenerative diseases.

Contribution

The first rational design of oligonucleotide chaperones targeting hnRNP A1 with high specificity and anti-aggregation activity.

Findings

The designed chaperone binds to hnRNP A1 at micromolar concentrations.

The chaperone inhibits hnRNP A1 condensate and fibril formation by over 90%.

Modifications to the oligonucleotide backbone improved its affinity and activity.

Abstract

Pharmacological chaperones of heterogeneous nuclear ribonucleoproteins (hnRNPs) show promise as potential neuroprotective drug candidates. They are expected to prevent the accumulation of neurotoxic hnRNP biocondensates and aggregates, which are hallmarks of severe degenerative diseases. Here, we present the first rational design of oligonucleotide chaperones of hnRNP A1. This design was inspired by previous studies on the specificity of the RNA recognition motif (RRM) and the RGG motif of hnRNP A1 for endogenous nucleic acids. To obtain robust and specific chaperones, we combined an RRM-binding sequence with an RGG-binding G-quadruplex oligonucleotide that inhibits hnRNP A1 aggregation and introduced various modifications into the sugar-phosphate backbone of the oligonucleotide. Modifications that locked the RRM-binding sequence in a conformational state characteristic of RNA improved chaperone affinity and activity. The former was assessed using microscale thermophoresis assays, while the latter was evaluated using fluorimetry and microscopy. The leading chaperone bound to hnRNP A1 at micromolar concentrations and inhibited the assembly of its condensates and amyloid-like aggregates (fibrils) by over 90%.

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