PET imaging of distinct brain uptake of a nanobody and similarly-sized PAMAM dendrimers after intra-arterial administration
- 168 Downloads
We have recently shown that intracerebral delivery of an anti-VEGF monoclonal antibody bevacizumab using an intra-arterial (IA) infusion is more effective than intravenous administration. While antibodies are quickly emerging as therapeutics, their disadvantages such as large size, production logistics and immunogenicity motivate search for alternatives. Thus we have studied brain uptake of nanobodies and polyamidoamine (PAMAM) dendrimers.
Nanobodies were conjugated with deferoxamine (DFO) to generate NB(DFO)2. Generation-4 PAMAM dendrimers were conjugated with DFO, and subsequently primary amines were capped with butane-1,2-diol functionalities to generate G4(DFO)3(Bdiol)110. Resulting conjugates were radiolabeled with zirconium-89. Brain uptake of 89ZrNB(DFO)2 and 89ZrG4(DFO)3(Bdiol)110 upon carotid artery vs tail vein infusions with intact BBB or osmotic blood–brain barrier opening (OBBBO) with mannitol in mice was monitored by dynamic positron emission tomography (PET) over 30 min to assess brain uptake and clearance, followed by whole-body PET-CT (computed tomography) imaging at 1 h and 24 h post-infusion (pi). Imaging results were subsequently validated by ex-vivo biodistribution.
Intravenous administration of 89ZrNB(DFO)2 and 89ZrG4(DFO)3(Bdiol)110 resulted in their negligible brain accumulation regardless of BBB status and timing of OBBBO. Intra-arterial (IA) administration of 89ZrNB(DFO)2 dramatically increased its brain uptake, which was further potentiated with prior OBBBO. Half of the initial brain uptake was retained after 24 h. In contrast, IA infusion of 89ZrG4(DFO)3(Bdiol)110 resulted in poor initial accumulation in the brain, with complete clearance within 1 h of administration. Ex-vivo biodistribution results reflected those on PET-CT.
IA delivery of nanobodies might be an attractive therapeutic platform for CNS disorders where prolonged intracranial retention is necessary.
KeywordsPET Nanobody Dendrimer Intra-arterial Zirconium Brain
This work was funded by National Institutes of Health (NIH) R01NS091110, R21NS106436, P41 EB024495.
Compliance with ethical standards
Disclosure of potential conflict of interest
JAG has a financial and/or business interests in Gulliver Biomed BVBA, a company that licensed the tested nanobody; however, since the nanobody does not have a brain target there are no direct benefits to Gulliver Biomed. The remaining authors declared no conflict of interest related to the current work.
All animal procedures were carried out under protocols approved by the Johns Hopkins Animal Care and Use Committee. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. This article does not contain any studies with human participants performed by any of the authors.
- 5.Boockvar JA, Tsiouris AJ, Hofstetter CP, Kovanlikaya I, Fralin S, Kesavabhotla K, et al. Safety and maximum tolerated dose of superselective intraarterial cerebral infusion of bevacizumab after osmotic blood–brain barrier disruption for recurrent malignant glioma. Clinical article. J Neurosurg. 2011;114:624–32. https://doi.org/10.3171/2010.9.JNS101223.CrossRefGoogle Scholar
- 6.Lesniak WG, Chu C, Jablonska A, Du Y, Pomper MG, Walczak P, et al. A distinct advantage to intraarterial delivery of 89zr-bevacizumab in pet imaging of mice with and without osmotic opening of the blood–brain barrier. J Nucl Med. 2019;60(5):617–22. https://doi.org/10.2967/jnumed.118.218792.CrossRefGoogle Scholar
- 8.Janowski M, Walczak P, Pearl MS. Predicting and optimizing the territory of blood–brain barrier opening by superselective intra-arterial cerebral infusion under dynamic susceptibility contrast MRI guidance. J Cereb Blood Flow Metab. 2016;36:569–75. https://doi.org/10.1177/0271678X15615875.CrossRefGoogle Scholar
- 9.Lyczek A, Arnold A, Zhang J, Campanelli JT, Janowski M, Bulte JW, et al. Transplanted human glial-restricted progenitors can rescue the survival of dysmyelinated mice independent of the production of mature, compact myelin. Exp Neurol. 2017;291:74–86. https://doi.org/10.1016/j.expneurol.2017.02.005.CrossRefGoogle Scholar
- 11.Ingram JR, Schmidt FI, Ploegh HL. Exploiting Nanobodies’ singular traits. Annu Rev Immunol. 2018;36:695–715. https://doi.org/10.1146/annurev-immunol-042617-053327.CrossRefGoogle Scholar
- 17.Zhang F, Mastorakos P, Mishra MK, Mangraviti A, Hwang L, Zhou J, et al. Uniform brain tumor distribution and tumor associated macrophage targeting of systemically administered dendrimers. Biomaterials. 2015;52:507–16. https://doi.org/10.1016/j.biomaterials.2015.02.053.CrossRefGoogle Scholar
- 20.Vosjan MJ, Perk LR, Visser GW, Budde M, Jurek P, Kiefer GE, et al. Conjugation and radiolabeling of monoclonal antibodies with zirconium-89 for PET imaging using the bifunctional chelate p-isothiocyanatobenzyl-desferrioxamine. Nat Protoc. 2010;5:739–43. https://doi.org/10.1038/nprot.2010.13.CrossRefGoogle Scholar
- 25.Zhang Y, Sun Y, Xu X, Zhang X, Zhu H, Huang L, et al. Synthesis, biodistribution, and microsingle photon emission computed tomography (SPECT) imaging study of technetium-99m labeled PEGylated dendrimer poly(amidoamine) (PAMAM)-folic acid conjugates. J Med Chem. 2010;53:3262–72. https://doi.org/10.1021/jm901910j.CrossRefGoogle Scholar
- 28.Gomes JR, Cabrito I, Soares HR, Costelha S, Teixeira A, Wittelsberger A, et al. Delivery of an anti-transthyretin Nanobody to the brain through intranasal administration reveals transthyretin expression and secretion by motor neurons. J Neurochem. 2018;145:393–408. https://doi.org/10.1111/jnc.14332.CrossRefGoogle Scholar
- 32.Bala G, Blykers A, Xavier C, Descamps B, Broisat A, Ghezzi C, et al. Targeting of vascular cell adhesion molecule-1 by 18F-labelled nanobodies for PET/CT imaging of inflamed atherosclerotic plaques. Eur Heart J Cardiovasc Imaging. 2016;17:1001–8. https://doi.org/10.1093/ehjci/jev346.CrossRefGoogle Scholar