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Aptamer-Functionalized Microbubbles Targeted to P-selectin for Ultrasound Molecular Imaging of Murine Bowel Inflammation

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Abstract

Purpose

Our objectives were to develop a targeted microbubble with an anti-P-selectin aptamer and assess its ability to detect bowel inflammation in two murine models of acute colitis.

Procedures.

Lipid-shelled microbubbles were prepared using mechanical agitation. A rapid copper-free click chemistry approach (azide-DBCO) was used to conjugate the fluorescent anti-P-selectin aptamer (Fluor-P-Ap) to the microbubble surface. Bowel inflammation was chemically induced using 2,4,6-trinitrobenzenesulfonic acid (TNBS) in both Balb/C and interleukin-10-deficient (IL-10 KO) mice. Mouse bowels were imaged using non-linear contrast mode following an i.v. bolus of 1 × 108 microbubbles. Each mouse received a bolus of aptamer-functionalized and non-targeted microbubbles. Mouse phenotypes and the presence of P-selectin were validated using histology and immunostaining, respectively.

Results

Microbubble labelling of Fluor-P-Ap was complete after 20 min at 37 ̊C. We estimate approximately 300,000 Fluor-P-Ap per microbubble and confirmed fluorescence using confocal microscopy. There was a significant increase in ultrasound molecular imaging signal from both Balb/C (p = 0.003) and IL-10 KO (p = 0.02) mice with inflamed bowels using aptamer-functionalized microbubbles in comparison to non-targeted microbubbles. There was no signal in healthy mice (p = 0.4051) using either microbubble.

Conclusions

We constructed an aptamer-functionalized microbubble specific for P-selectin using a clinically relevant azide-DBCO click reaction, which could detect bowel inflammation in vivo. Aptamers have potential as a next generation targeting agent for developing cost-efficient and clinically translatable targeted microbubbles.

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Acknowledgements

We would like to acknowledge that BioRender.com was used to create the Fig. 1.

Funding

This work was supported by a Saskatchewan Health Research Foundation Establishment Grant.

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Authors and Affiliations

  1. Department of Medical Imaging, University of Saskatchewan, Saskatoon, SK, S7N 5E5, Canada

    Una Goncin & Steven Machtaler

  2. Department of Electrical and Software Engineering, Schulich School of Engineering, University of Calgary, Calgary, AB, T2N 4V8, Canada

    Laura Curiel

  3. Department of Pathology and Laboratory Medicine, University of Saskatchewan, Saskatoon, SK, S7N 5E5, Canada

    C. Ronald Geyer

Authors
  1. Una Goncin
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  2. Laura Curiel
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  3. C. Ronald Geyer
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  4. Steven Machtaler
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Contributions

Una Goncin: resources, conception, investigation, formal analysis, interpretation, writing — original draft, writing — review & editing, approval. Laura Curiel: investigation, interpretation, writing, review. C Ronald Geyer: resources/acquisition, conception, writing — review & editing, approval. Steven Machtaler: resources, conception, interpretation, writing — review & editing, approval.

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Correspondence to Steven Machtaler.

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Goncin, U., Curiel, L., Geyer, C.R. et al. Aptamer-Functionalized Microbubbles Targeted to P-selectin for Ultrasound Molecular Imaging of Murine Bowel Inflammation. Mol Imaging Biol 25, 283–293 (2023). https://doi.org/10.1007/s11307-022-01755-9

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