Zhi Zuo1,2,3, Tatiana Syrovets4, Tao Wang5, Yu-zhou Wu5, Felicitas Genze4, Alireza Abaei3, Ina Vernikouskaya2, Wei-na Liu5, Gen-shan Ma1, Tanja Weil5, Thomas Simmet4, and Volker Rasche2,3
1Department of Cardiology, Zhongda Hospital, Medical School of Southeast University, Nanjing, China, People's Republic of, 2Department of Internal Medicine II, University Hospital Ulm, Ulm, Germany, 3Core Facility Small Animal MRI, Medical Faculty, Ulm University, Ulm, Germany, 4Institute of Pharmacology of Natural Products and Clinical Pharmacology, Ulm University, Ulm, Germany, 5Institute of Organic Chemistry III, Ulm University, Ulm, Germany
Synopsis
High-resolution MRI and chicken chorioallantoic membrane (CAM) cancer xenografts model are employed for the assessment of bio-distribution of polypeptide hybrid biomaterials cHSA-PEO (2000)16-Gd. The breast carcinomas were planted on the CAM, which were intravascular injected with cHSA-PEO (2000)16-Gd, gadofosveset (Vasovist®) and gadobenate dimeglumine (Multihance®) before MR scanning. The detailed internal structures of tiny cancer were clearly highlighted, which was proven by the immune-histological analysis. Assessment of the bio-distribution of contrast agents renders feasible. The cHSA-PEO (2000)16-Gd has exhibited an improved specific tissue uptake by cancer cell xenografts and long lasting enhancement compared to traditional contrast agents.Introduction
As immune-deficient in vivo model, the CAM
model is much cheaper, easier for tumor implantation compared to nude rodent
model and has no ethical or legal concerns referring to European and USA experimental
animal protection laws. It has gained great interest for quality testing of
novel contrast agent (CA) materials designed for the early stage cancer diagnosis
1. With the recent developments in immobilization of the chicken
embryo
2, non-invasive high-resolution imaging of the embryos and cancers
planted on the CAM has rendered feasible. In this contribution it is investigated
whether the CAM model could be used for the MRI assessment of a self-assembled
Gd-DOTA conjugated protein based polypeptide copolymer cHSA-PEO (2000)
16-Gd.
Methods and Materials
28 chicken embryos with planted mammalian carcinoma
were investigated. The carcinomas were planted at day 7 after embryo incubation.
Carcinomas grew for 9 days and imaging was performed at day 16. Prior to
imaging, the embryos were cooled at 4℃
for 110 minutes for immobilization. All scanning was performed by a T
1-weighted
three-dimensional FLASH sequence with acquisition parameters as follows: TR/TE
= 6/2 ms, matrix = 400×439×96, spatial resolution = 100×100×560 µm³ and NSA =
2. Eight chicken embryos were injected with different gadofosveset doses (5, 10,
20, 50, 60, 70, 80, and 90μl 0.5mmol/ml gadofosveset solution) for assessment
of the optimal contrast agent dosage. Scanning was performed right after
injection and the optimal dosage identified by analysis of the signal-to-noise
ratio (SNR) in the umbilical vein. After optimal gadofosveset dosage was
confirmed, 10 chick embryos were used for the assessment of the gadofosveset
bio-distribution systemically injection. All embryos were scanned prior to, 30min,
3h, 20h and 40h after intravenous administration of CA. SNR data of different
organs (vessels, allantoic fluid, liver, and brain) of chick embryos at
different time points after contrast agent injection were measured and
calculated. After all protocols were confirmed, novel MR contrast agent
material was assessed and compared with gadofosveset and gadobenate dimeglumine.
Results
MR images of post-injection embryos with different CA
dose and resulting dose-SNR curves are provided in
Figure 1. A dose of 80μl was
chosen as optimal gadofosveset dosage for day 16 chicken embryos. Example MR images
of gadofosveset bio-distribution and change of the mean SNR ± std. Dev in
different embryo organs are provided in
Figure 2. The bio-distribution of the
injected gadofosveset can be clearly visualized with high spatial fidelity in
ovo. From Figure 2 an early enhancement of the blood circulation can be
appreciated followed by enhancement the liver parenchyma. Finally the allantoic
fluid is enhanced. The enhancement variability in cancer cells xenograft over
40 hours after Vasovist intravenous injection and whole egg enhancement at 0.5
hour time point is shown in
Figure 3. The cancer cells xenograft was enhanced
almost from the same time with the blood pool. The correlation of immune-histological
analysis of tumor xenografts and MR image is shown in
Figure 4. A high
correlation between the MR images and immune-histological analysis can be
appreciated. The comparison of cancer xenograft enhancement by novel material
and two traditional commercial CA is shown in
Figure 5. Only in samples
injected with cHSA-PEO (2000)
16-Gd the enhancement is restricted to the cancer
cells xenograft with persistent enhancement of up to 20 hours after injection.
Discussion and Conclusion
To our knowledge, this is the first report on the MR assessment of novel CA in CAM model with cancer cell xenograft with intravenous administration. The novel CA material cHSA-PEO (2000)
16-Gd has exhibited an improved uptake by cancer cell xenografts and long lasting enhancement compared to traditional CA. With the protocol introduced, the CAM model presented great potential of initial testing of novel CA material thus maybe replacing some animal experiments.
Acknowledgements
No acknowledgement found.References
1. Vargas A, Zeisser-Labouèbe M, Lange N, et al. The chick embryo and its chorioallantoic membrane (CAM) for the in vivo evaluation of drug delivery systems. Adv Drug Deliv Rev 2007 Sep 30;59(11):1162-76.
2. Zuo Z, Syrovets T, Genze F, et al. High-resolution MRI analysis of breast cancer xenograft on the chick chorioallantoic membrane. NMR Biomed 2015 Apr;28(4):440-7.