IN VIVO HYPERCEST DETECTION OF CUCURBIT[6]URIL IN RAT ABDOMEN
Francis Hane1, Tao Li1, Peter Smylie1, and Mitchell S Albert1

1Lakehead University, Thunder Bay, ON, Canada

Synopsis

We used the MRI HyperCEST technique to detect the presence of the xenon encapsulating cage molecule cucurbit[6]uril (CB6) in the abdomen of a rat. We believe that this is the first in vivo demonstration of a xenon based biosensor. We were able to observe a HyperCEST signal depletion of 53% within the intraperitoneal space of the rat. Our results demonstrate the feasibility of HyperCEST biosensors to move from in vitro to in vivo studies.

Audience & Purpose

Hyperpolarized (HP) Xenon-129 MRI biosensors have the potential to detect pathological molecules within the body at extremely low concentrations with MRI-like resolution and Positron Emission Tomography (PET)-like sensitivity. Combining the increased sensitivity of hyperpolarization with the Hyperpolarized Chemical Exchange Saturation Transfer (HyperCEST) technique can provide a sensitivity increase of a billion times over thermally polarized xenon. In this work we present the first in vivo HyperCEST spectra of cucurbit[6]uril (CB6) HyperCEST contrast agent in a live animal. This work is important for other researchers in the field in translating in vitro testing of HP gas biosensors to preclinical testing and thence to clinical trials.

Methods

A 10 mL 5.0 mM solution of CB6 was prepared in a PBS solvent. 83.6% isotopically enriched 129Xe was hyperpolarized to 30% using a Xemed Xebox polarizer (Durham, NH). 5 mL of HP 129Xe was introduced into the CB6 solution and shaken for 12 seconds. 5 mL of the CB6 solution containing HP 129Xe was injected into the intraperitoneal (IP) space of ~400 g male Sprague-Dawley rats. The rats were placed into a custom RF coil tuned to a Larmor frequency of 129Xe (35.33 MHz) at 3 T. A HyperCEST pulse sequence (16- 6 ms 3-lobe sinc pulse, 3 ms pulse interval) was applied at the CB6-Xe resonance frequency (+124.3 ppm). NMR spectra were acquired with both off-resonance control pulses and on-resonance saturation pulses. MR images were taken with 150 mm x 150 mm FOV, 32x32 voxels, 300 mm slice thickness. TR=197 ms, TE=0.77 ms.

Results & Discussion

We observed a 53% HyperCEST depletion of the CB6-xenon complex following the applied saturation pre-pulse. We observed a signal peak at +192.4 ppm with respect to the xenon gas phase peak corresponding to xenon dissolved in plasma [1]. We observed a shoulder down field from the peak (+124.3 ppm) which we assume to be caused by the CB6/Xe complex. However, given the complex interactions and exchange broadening in vivo, this peak was not independently resolvable.

Conclusions

We believe that these data are the first report of in vivo application of a HP Xe based MRI biosensor in a live animal model. We will continue our work by obtaining HyperCEST saturation maps in live animals to detect in vivo pathological molecules of interest.

Acknowledgements

FH would like to acknowledge the generous support of the donors of the BrightFocus Foundation.

References

1. Wang Y, Dmochowski I. Cucurbit[6]uril is an ultrasensitive 129Xe NMR contrast agent. Chem. Comm. 2015; 51:8982–8985.

Figures

(Left) Free Induction Decay (FID) spectrum centered on the intraperitoneal (IP) space of a Sprague-Dawley rat following injection of 5 mL of CB6 dissolved in PBS combined with HP Xe. An off-resonant HyperCEST pulse was applied at + 259 ppm. (Right) Similar to left but using an on-resonance (+124.3 ppm) HyperCEST saturation pulse.

A 1H MRI of the abdomen of a Sprague-Dawley rat overlaid with a HyperCEST saturation map showing a HyperCEST signal from 5 mL of 5 mM CB6 solution injected into the rat intraperitoneally.



Proc. Intl. Soc. Mag. Reson. Med. 24 (2016)
0814