Endosomal trafficking regulates receptor-mediated transcytosis of antibodies across the blood brain barrier

Haqqani, Arsalan S.; Delaney, Christie E.; Brunette, Eric; Baumann, Ewa; Farrington, Graham K.; Sisk, William P.; Eldredge, John; Ding, Wen; Tremblay, Tammy-Lynn; Stanimirovic, Danica

doi:10.1177/0271678x17740031

Cited by 67 publications

(78 citation statements)

References 49 publications

(113 reference statements)

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“…Recently, we have described gradient fractionation and quantitative protein characterization of subcellular fractions of SV‐ARBEC using nanoLC‐SRM (Haqqani et al . ). The low‐density fractions (LDF 2‐4) contained markers of late endosomes/lysosomes (Lamp 1/2, rM6pr, Rab7a, Rab11a/b), whereas high‐density fractions (HDF 5‐7) were enriched in markers of early and recycling endosomes (Rab5a, Eaa1) [(Haqqani et al .…”

Section: Resultsmentioning

confidence: 97%

“…The low‐density fractions (LDF 2‐4) contained markers of late endosomes/lysosomes (Lamp 1/2, rM6pr, Rab7a, Rab11a/b), whereas high‐density fractions (HDF 5‐7) were enriched in markers of early and recycling endosomes (Rab5a, Eaa1) [(Haqqani et al . ); Table 3]; very high‐density fractions (VHDF 8‐10) exhibited unique profile of markers, similar to some subpopulations of microvesicular bodies (Haqqani et al . ).…”

Section: Resultsmentioning

confidence: 99%

“…Endosome isolation and characterization was performed as described recently (Haqqani et al . ). SV‐ARBEC were grown to confluency on rat tail collagen type I‐coated plastic dishes, as described previously (Garberg et al .…”

Section: Methodsmentioning

confidence: 97%

“…The enrichment of various molecular markers in isolated fractions was evaluated using western blot and mass spectrometry as recently described (Haqqani et al . ). These studies identified low ‐ density fractions 2‐4 (LDFs) as late endosomes and lysosomes, high‐density fractions 4‐7 (HDFs) as early and recycling endosomes, and very high‐density fractions 8‐10 (VHDFs) as a subset of multivesicular bodies.…”

Section: Methodsmentioning

confidence: 97%

“…OX26 affinity variants and control antibody levels in samples from in vitro BBB model and in endosomal cell fractions were trypsin digested (see above) and quantified using targeted nanoLC MS/ MS on nanoAcquity UPLC (Waters, Milford, MA, USA) coupled to ESI LTQ XL ETD mass spectrometer (ThermoFisher) as previously described (Haqqani et al . ). Briefly, the samples were injected onto a PepMap100, 5 μm 100 angstrom C18 trap (160454, ThermoFisher, Waltham, MA, USA) followed by eluting onto a 100 μm I.D.…”

Section: Methodsmentioning

confidence: 97%

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Intracellular sorting and transcytosis of the rat transferrin receptor antibody OX26 across the blood–brain barrier in vitro is dependent on its binding affinity

Haqqani

Thom²,

Burrell³

et al. 2018

Journal of Neurochemistry

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The blood–brain barrier (BBB) is a formidable obstacle to the delivery of therapeutics to the brain. Antibodies that bind transferrin receptor (TfR), which is enriched in brain endothelial cells, have been shown to cross the BBB and are being developed as fusion proteins to deliver therapeutic cargos to brain targets. Various antibodies have been developed for this purpose and their in vivo evaluation demonstrated that either low affinity or monovalent receptor binding re‐directs their transcellular trafficking away from lysosomal degradation and toward improved exocytosis on the abluminal side of the BBB. However, these studies have been performed with antibodies that recognize different TfR epitopes and have different binding characteristics, preventing inter‐study comparisons. In this study, the efficiency of transcytosis in vitro and intracellular trafficking in endosomal compartments were evaluated in an in vitro BBB model for affinity variants (K d from 5 to174 nM) of the rat TfR‐binding antibody, OX26. Distribution in subcellular fractions of the rat brain endothelial cells was determined using both targeted quantitative proteomics‐selected reaction monitoring and fluorescent imaging with markers of early‐ and late endosomes. The OX26 variants with affinities of 76 and 108 nM showed improved trancytosis (Papp values) across the in vitro BBB model compared with a 5 nM OX26. Although ~40% of the 5 nM OX26 and ~35% of TfR co‐localized with late‐endosome/lysosome compartment, 76 and 108 nM affinity variants showed lower amounts in lysosomes and a predominant co‐localization with early endosome markers. The study links bivalent TfR antibody affinity to mechanisms of sorting and trafficking away from late endosomes and lysosomes, resulting in improvement in their transcytosis efficiency.Open Practices Open Science: This manuscript was awarded with the Open Materials Badge.For more information see: https://cos.io/our-services/open-science-badges/ Cover Image for this issue: doi: 10.1111/jnc.14193.

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Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 97%

Section: Methodsmentioning

confidence: 97%

Section: Methodsmentioning

confidence: 97%

See 3 more Smart Citations

Intracellular sorting and transcytosis of the rat transferrin receptor antibody OX26 across the blood–brain barrier in vitro is dependent on its binding affinity

Haqqani

Thom²,

Burrell³

et al. 2018

Journal of Neurochemistry

Self Cite

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Multifunctional Nanotheranostics for Overcoming the Blood–Brain Barrier

Beygi,

Oroojalian,

Azizi‐Arani

et al. 2024

Adv Funct Materials

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The blood–brain barrier (BBB) is a tailored system of capillary endothelial cells intermixed with tight junctions and adherent junctions that regulates the transport of various materials and substances between the blood vasculature and the central nervous system (CNS). However, in cases of brain diseases, BBB's protective and regulatory effects hamper therapeutics from reaching affected sites in sufficient quantities. This has so far been a leading challenge in treating CNS diseases and disorders. For this problem to be overcome, recent research has sought to develop novel modalities to achieve efficient therapy and alleviate associated symptoms. Therefore, numerous strategies have operated in recent years to address the limitations of traditional and invasive methods, including poor brain penetration and serious side effects. As a desperately in‐demand technology, nanotheranostics has particularly shown promising results. Herein, this review reports recent advancements in CNS nanotheranostics and novel techniques and nanotechnology‐based strategies developed for treating neurodegenerative disorders. The study provides comprehensive data on the subject to be used for future studies for the therapy and management of CNS disorders and diseases.

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Overcoming the Blood–Brain Barrier: The Role of Nanomaterials in Treating Neurological Diseases

et al. 2018

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Therapies directed toward the central nervous system remain difficult to translate into improved clinical outcomes. This is largely due to the blood-brain barrier (BBB), arguably the most tightly regulated interface in the human body, which routinely excludes most therapeutics. Advances in the engineering of nanomaterials and their application in biomedicine (i.e., nanomedicine) are enabling new strategies that have the potential to help improve our understanding and treatment of neurological diseases. Herein, the various mechanisms by which therapeutics can be delivered to the brain are examined and key challenges facing translation of this research from benchtop to bedside are highlighted. Following a contextual overview of the BBB anatomy and physiology in both healthy and diseased states, relevant therapeutic strategies for bypassing and crossing the BBB are discussed. The focus here is especially on nanomaterial-based drug delivery systems and the potential of these to overcome the biological challenges imposed by the BBB. Finally, disease-targeting strategies and clearance mechanisms are explored. The objective is to provide the diverse range of researchers active in the field (e.g., material scientists, chemists, engineers, neuroscientists, and clinicians) with an easily accessible guide to the key opportunities and challenges currently facing the nanomaterial-mediated treatment of neurological diseases.

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Endosomal trafficking regulates receptor-mediated transcytosis of antibodies across the blood brain barrier

Cited by 67 publications

References 49 publications

Intracellular sorting and transcytosis of the rat transferrin receptor antibody OX26 across the blood–brain barrier in vitro is dependent on its binding affinity

Intracellular sorting and transcytosis of the rat transferrin receptor antibody OX26 across the blood–brain barrier in vitro is dependent on its binding affinity

Multifunctional Nanotheranostics for Overcoming the Blood–Brain Barrier

Overcoming the Blood–Brain Barrier: The Role of Nanomaterials in Treating Neurological Diseases

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