Oligonucleotides in PET imaging

Nucleotides are the build blocks of DNA and RNA, consisting of a nucleobase (adenine, cytosine, guanine, thymine, or uracil), a 5-carbon sugar, and a phosphate group. Oligonucleotide can hybridize to another chain if their sequences can form G☰C and A⚌T or A⚌U pairs. Thus, oligonucleotides can be used as ligands for DNA and RNA (antisense oligonucleotides). Oligonucleotides can also be designed to specifically bind to other molecular targets (aptamer oligonucleotides). Techniques for generic labelling and protection from nucleases are available, but directing oligonucleotides into the cell cytoplasm and nucleus while still maintaining their functionality is still challenging.

In blood plasma, oligonucleotides are bound to albumin (Srinivasan et al., 1995), which is common to negatively charged molecules. Oligonucleotides tend to bind to other proteins, too, and while this may not be a problem for medicinal use of oligonucleotide, it reduces image quality by directly increasing the nonspecific binding and indirectly by limiting glomerular filtration and urinary excretion, increasing the halflife of the radiopharmaceutical in the plasma.

Antisense oligonucleotides

Antisense single-stranded oligonucleotides (ASOs) bind to its target RNA, and inhibit its translation. The secondary structures of RNAs (hairpins, stem-loops) may prevent or slow down the hybrid formation with the antisense oligonucleotide. The number of mRNAs copies in a cell may be too low for detection in vivo. Antisense imaging may therefore be most successful in detecting viral or bacterial mRNAs that are abundant in infected tissue.

Aptamer oligonucleotides

DNA and RNA aptamers (usually 20-100 bases) are usually single-stranded, and can form 3D-structures, enabling them to bind to a specific target (aptatope); targets can be anything from small ions such as Zn2+ to viruses and bacteria. Libraries of oligonucleotides can be synthesized randomly and simultaneously screened for their binding affinity to a target molecule. This procedure is easier and faster than the methods for production of monoclonal antibodies, and it works with targets that are toxic or non-immunogenic. Aptamers are an order of magnitude smaller than antibodies (10-20 kDa versus 150 kDa), and therefore have faster endothelial passage and clearance. Aptamers, like antibodies, are usually targeted against molecules that are abundant enough for successful detection with the excellent sensitivity of modern PET devices.

Escort aptamers can be used for targeted delivery of drugs, and diagnostic or therapeutic radionuclides.

Riboswitches are naturally occurring aptamers, mainly found in the untranslated regions of mRNA molecules; when bound to their target molecule, the 3D structural change in mRNA affects its translational activity.

Promising aptamer radiopharmaceuticals have been developed for example for tumour PET imaging, targeting EGFR (HER1) (Cheng et al., 2019) and PTK7 (Jacobson et al., 2015).


MicroRNAs (miRNAs) are endogenous single-stranded non-coding RNAs consisting of 16-23 nucleotides. MicroRNAs regulate gene expression by inhibiting the post-transcriptional process: miRNA pairs with messenger RNA (mRNA) to its untranslated regions, which leads to formation of miRNA-induced silencing complex (miRISC) that includes ribonucleases. If the sequences of miRNA and mRNA do not match well, the miRISC merely inhibits the translation of that mRNA, but when the match is good, then miRISC starts RNA degradation process (RNA interference, RNAi) (Hernandez et al., 2013). Thousands of miRNAs have been identified. Certain miRNA (for instance miR-21) are often over-expressed or down-regulated in certain tumour cells. miR-15b is important for tissue remodelling, especially in the bones. Short interfering RNAs (siRNAs) function similarly, but are exogenous double-stranded RNAs. Pre-miRNAs are transcribed in the nucleus, and digested into mature miRNAs in the cytoplasm (Mäkilä et al., 2019).

miRNAs and siRNAs are being tested as therapeutic agents, but their short in vivo halflife, low cellular uptake (via endocytosis), and off-target effects remain a problem. Pharmacokinetics, biodistribution, and specific tissue uptake of positron-emitting radionuclide labelled miRNAs with PET (Mäkilä et al., 2019).

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Updated at: 2019-03-18
Created at: 2017-10-28
Written by: Vesa Oikonen