Grabody B, an IGF1 receptor-based shuttle, mediates efficient delivery of biologics across the blood-brain barrier

Shin, Jung-Won; An, Sungwon; Kim, Dongin; Kim, Hyun‐Joo; Ahn, Jin-Hyung; Eom, Jaehyun; You, Weon‐Kyoo; Yun, Hyo-In; Lee, Bora; Sung, Byungje; Jung, Jinwon; Kim, Sehyun; Son, Yong-Gyu; Sung, Eun-Sil; Lee, Hanbyul; Lee, Su-Yeon; Song, Daehae; Pak, Youngdon; Sandhu, Jagdeep K.; Haqqani, Arsalan S.; Stanimirovic, Danica; Yoo, Jiseon; Kim, Dong Hwan; Maeng, Sungho; Lee, Jeonghun; Lee, Sang Hoon

doi:10.1016/j.crmeth.2022.100338

Cited by 13 publications

(21 citation statements)

References 70 publications

(116 reference statements)

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“…GO analysis of the genes in clusters showing higher expression levels with increasing culture time and clusters where gene expression in BBB was higher than in BMEC only revealed many genes related to cellular migration as well as ECM and vascular remodelling, indicating astrocytes and pericytes are involved in modulating the BBB. Additionally, insulin-like growth factor (IGF) binding protein, which is an important transport system utilized by drug development researchers to deliver biologics to the brain 37 – 39 , was also more highly expressed in BBB compared to BMEC only, suggesting that astrocytes and pericytes may be required for the expression of some transporter proteins in the BBB.…”

Section: Resultsmentioning

confidence: 99%

Modular tissue-in-a-CUBE platform to model blood-brain barrier (BBB) and brain interaction

Koh,

Hagiwara

2024

Commun Biol

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With the advent of increasingly sophisticated organoids, there is growing demand for technology to replicate the interactions between multiple tissues or organs. This is challenging to achieve, however, due to the varying culture conditions of the different cell types that make up each tissue. Current methods often require complicated microfluidic setups, but fragile tissue samples tend not to fare well with rough handling. Furthermore, the more complicated the human system to be replicated, the more difficult the model becomes to operate. Here, we present the development of a multi-tissue chip platform that takes advantage of the modularity and convenient handling ability of a CUBE device. We first developed a blood-brain barrier-in-a-CUBE by layering astrocytes, pericytes, and brain microvascular endothelial cells in the CUBE, and confirmed the expression and function of important tight junction and transporter proteins in the blood-brain barrier model. Then, we demonstrated the application of integrating Tissue-in-a-CUBE with a chip in simulating the in vitro testing of the permeability of a drug through the blood-brain barrier to the brain and its effect on treating the glioblastoma brain cancer model. We anticipate that this platform can be adapted for use with organoids to build complex human systems in vitro by the combination of multiple simple CUBE units.

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Section: Resultsmentioning

confidence: 99%

Modular tissue-in-a-CUBE platform to model blood-brain barrier (BBB) and brain interaction

Koh,

Hagiwara

2024

Commun Biol

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“…The nanobody constructs developed here have relatively fast kinetics and may be potentially useful for short-acting therapeutics and diagnostic applications. However, for applications where a long duration of action is favorable such as treatment of chronic brain diseases, solutions based on receptor mediated transcytosis using conventional immunoglobulins may be preferred [6] [8] [12] [13] [30] [31] [32]. It may be possible to further prolong the duration of action of nanobody-based constructs by making hybrid nanobody-IgG fusions, conjugation to polyethylene glycol, or other approaches, but these may negate many of the advantages of nanobody-based constructs noted elsewhere.…”

Section: Supplemental Discussionmentioning

confidence: 99%

“…The first clinically approved therapeutic employing a receptor mediated transcytosis BBB shuttle, pabinafusp alfa, consists of a TfR-binding monoclonal antibody fused to the enzyme iduronate-2-sulfatase, which is defective in mucopolysaccharidosis type II (Hunter syndrome) [6] [7]. Additional targets for BBB transcytosis shuttles include the insulin receptor [8] [9], IGF1 receptor [10] [11] [12], CD98hc [13] [14], TMEM30 [15], and others [1] [2] [16].…”

Section: Introductionmentioning

confidence: 99%

Enhancedin VivoBlood Brain Barrier Transcytosis of Macromolecular Cargo Using an Engineered pH-sensitive Mouse Transferrin Receptor Binding Nanobody

Esparza

Francescutti

et al. 2023

Preprint

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Background: The blood brain barrier limits entry of macromolecular diagnostic and therapeutic cargos. Blood brain barrier transcytosis via receptor mediated transport systems, such as the transferrin receptor, can be used to carry macromolecular cargos with variable efficiency. Transcytosis involves trafficking through acidified intracellular vesicles, but it is not known whether pH-dependent unbinding of transport shuttles can be used to improve blood brain barrier transport efficiency. Methods: A mouse transferrin receptor binding nanobody, NIH-mTfR-M1, was engineered to confer greater unbinding at pH 5.5 vs 7.4 by introducing multiple histidine mutations. The histidine mutant nanobodies were coupled to neurotensin for in vivo functional blood brain barrier transcytosis testing via central neurotensin-mediated hypothermia in wild-type mice. Multi-nanobody constructs including the mutant M1R56H, P96H, Y102H and two copies of the P2X7 receptor-binding 13A7 nanobody were produced to test proof-of-concept macromolecular cargo transport in vivo using quantitatively verified capillary depleted brain lysates and in situ histology. Results: The most effective histidine mutant, M1R56H, P96H, Y102H -neurotensin, caused >8 degrees C hypothermia after 25 nmol/kg intravenous injection. Levels of the heterotrimeric construct M1R56H, P96H, Y102H -13A7-13A7 in capillary depleted brain lysates peaked at 1 hour and were 60% retained at 8 hours. A control construct with no brain targets was only 15% retained at 8 hours. Addition of the albumin-binding Nb80 nanobody to make M1R56H, P96H, Y102H -13A7-13A7-Nb80 extended blood half-life from 21 minutes to 2.6 hours. At 30-60 minutes, biotinylated M1R56H, P96H, Y102H -13A7-13A7-Nb80 was visualized in capillaries using in situ histochemistry, whereas at 2-16 hours it was detected in diffuse hippocampal and cortical cellular structures. Levels of M1R56H, P96H, Y102H-13A7-13A7-Nb80 reached more than 3.5 percent injected dose/gram of brain tissue after 30 nmol/kg intravenous injection. However, higher injected concentrations did not result in higher brain levels, compatible with saturation and an apparent substrate inhibitory effect. Conclusion: The pH-sensitive mouse transferrin receptor binding nanobody M1R56H, P96H, Y102H may be a useful tool for rapid and efficient modular transport of diagnostic and therapeutic macromolecular cargos across the blood brain barrier in mouse models. Additional development will be required to determine whether this nanobody-based shuttle system will be useful for imaging and fast-acting therapeutic applications.

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“…In addition to the endogenous peptide, these BBB receptors enable the RMT of certain peptidomimetic MAbs, which bind exofacial epitopes on the receptor, and this binding causes RMT of the MAb across the BBB in parallel with the BBB transport of the endogenous peptide. The RMT of a receptor-specific MAb was demonstrated for a MAb against the TfR, designated a TfRMAb ( Pardridge et al, 1991 ), for a MAb against the human IR (HIR), designated a HIRMAb ( Pardridge et al, 1995 ), and a MAb against the IGFR, designated the IGFRMAb ( Shin et al, 2022 ; Yogi et al, 2022 ). Early work showed that biologics, which normally do not cross the BBB, were enabled to penetrate the BBB and induce in vivo CNS pharmacologic effects following intravenous (IV) administration providing the biologic was conjugated to a TfRMAb.…”

Section: Introductionmentioning

confidence: 99%

Treatment of Parkinson’s disease with biologics that penetrate the blood–brain barrier via receptor-mediated transport

Pardridge

2023

Front. Aging Neurosci.

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Parkinson’s disease (PD) is characterized by neurodegeneration of nigral-striatal neurons in parallel with the formation of intra-neuronal α-synuclein aggregates, and these processes are exacerbated by neuro-inflammation. All 3 components of PD pathology are potentially treatable with biologics. Neurotrophins, such as glial derived neurotrophic factor or erythropoietin, can promote neural repair. Therapeutic antibodies can lead to disaggregation of α-synuclein neuronal inclusions. Decoy receptors can block the activity of pro-inflammatory cytokines in brain. However, these biologic drugs do not cross the blood–brain barrier (BBB). Biologics can be made transportable through the BBB following the re-engineering of the biologic as an IgG fusion protein, where the IgG domain targets an endogenous receptor-mediated transcytosis (RMT) system within the BBB, such as the insulin receptor or transferrin receptor. The receptor-specific antibody domain of the fusion protein acts as a molecular Trojan horse to ferry the biologic into brain via the BBB RMT pathway. This review describes the re-engineering of all 3 classes of biologics (neurotrophins, decoy receptor, therapeutic antibodies) for BBB delivery and treatment of PD. Targeting the RMT pathway at the BBB also enables non-viral gene therapy of PD using lipid nanoparticles (LNP) encapsulated with plasmid DNA encoding therapeutic genes. The surface of the lipid nanoparticle is conjugated with a receptor-specific IgG that triggers RMT of the LNP across the BBB in vivo.

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Grabody B, an IGF1 receptor-based shuttle, mediates efficient delivery of biologics across the blood-brain barrier

Cited by 13 publications

References 70 publications

Modular tissue-in-a-CUBE platform to model blood-brain barrier (BBB) and brain interaction

Modular tissue-in-a-CUBE platform to model blood-brain barrier (BBB) and brain interaction

Enhancedin VivoBlood Brain Barrier Transcytosis of Macromolecular Cargo Using an Engineered pH-sensitive Mouse Transferrin Receptor Binding Nanobody

Treatment of Parkinson’s disease with biologics that penetrate the blood–brain barrier via receptor-mediated transport

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