Targeting Transferrin Receptor to Transport Antisense Oligonucleotides Across the Blood-Brain Barrier

Barker, Scarlett J; Thayer, Mai B.; Kim, Chaeyoung; Tatarakis, David; Simon, Matthew; Dial, Rebekah; Nilewski, Christian; Wells, Robert C.; Zhou, Yinhan; Afetian, Megan E.; Chappell, Alfred E.; Chew, Kylie S.; Chow, Johann; Clemens, Allisa; Discenza, Claire B; Dugas, Jason C.; Dwyer, Chrissa A.; Earr, Timothy; Ha, Connie; Huynh, David; Jayaraman, Srini; Kwan, Wanda; Mahon, Cathal; Pizzo, Michelle E.; Roche, Elysia; Sanders, Laura; Stergioulis, Alexander; Tong, Raymond K.; Tran, Hai; Zuchero, Joy Yu; Estrada, Anthony A.; Gadkar, Kapil; Koth, Christopher MM; Sanchez, Pascal; Thorne, Robert G.; Watts, Ryan J.; Sandmann, Thomas; Kane, Lesley A.; Rigo, Frank; Dennis, Mark S.; Lewcock, Joseph W.; DeVos, Sarah L.

doi:10.1101/2023.04.25.538145

Cited by 10 publications

(7 citation statements)

References 69 publications

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“…The transport of oligonucleotide therapeutics such as ASOs across the blood–brain-barrier by Brainshuttle-mediated transcytosis has been confirmed as a promising approach for the delivery of ASOs to the CNS. As a recent example, Barker et al have demonstrated that brain concentrations of up to 30 nM of a Malat1 ASO after four IV injections in nonhuman primates (NHP) can be achieved …”

Section: Resultsmentioning

confidence: 99%

“…For example, the need to escape a lysosomal localization may reduce access to the target mRNA in the nucleus. While this could have general implications for the conjugate approach, in the in vivo setting, the presence of the BBB necessitates a Brainshuttle for targeted transport across this barrier as the ASO alone is not able to cross it in sufficient amounts …”

Section: Resultsmentioning

confidence: 99%

“…Enabling a peripheral route of administration for ASOs for the treatment of neurological disorders holds great promise to unlock the full potential of this modality. The principle of using an antibody that has the ability to cross the BBB as a transport carrier has been successfully demonstrated before . Nevertheless, the design space of such a conjugate is vast, and single variations such as the conjugation site or linker chemistries can make the difference between a mere tool molecule and a drug candidate.…”

Section: Discussionmentioning

confidence: 99%

“…The principle of using an antibody that has the ability to cross the BBB as a transport carrier has been successfully demonstrated before. 79 Nevertheless, the design space of such a conjugate is vast, and single variations such as the conjugation site or linker chemistries can make the difference between a mere tool molecule and a drug candidate. In this work, we sought to create antibody-ASO conjugates with optimized PK properties via a diligent approach with the aim of establishing a blueprint for future efforts in the field.…”

Section: ■ Conclusionmentioning

confidence: 99%

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Diligent Design Enables Antibody-ASO Conjugates with Optimal Pharmacokinetic Properties

Sela,

Mansø,

Siegel

et al. 2023

Bioconjugate Chem.

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Antisense-oligonucleotides (ASOs) are a promising drug modality for the treatment of neurological disorders, but the currently established route of administration via intrathecal delivery is a major limitation to its broader clinical application. An attractive alternative is the conjugation of the ASO to an antibody that facilitates access to the central nervous system (CNS) after peripheral application and target engagement at the blood–brain barrier, followed by transcytosis. Here, we show that the diligent conjugate design of Brainshuttle-ASO conjugates is the key to generating promising delivery vehicles and thereby establishing design principles to create optimized molecules with drug-like properties. An innovative site-specific transglutaminase-based conjugation technology was chosen and optimized in a stepwise process to identify the best-suited conjugation site, tags, reaction conditions, and linker design. The overall conjugation performance was found to be specifically governed by the choice of buffer conditions and the structure of the linker. The combination of the peptide tags YRYRQ and RYESK was chosen, showing high conjugation fidelity. Elaborate conjugate analysis revealed that one leading differentiating factor was hydrophobicity. The increase of hydrophobicity by the ASO payload could be mitigated by the appropriate choice of conjugation site and the heavy chain position 297 proved to be the most optimal. Evaluating the properties of the linker suggested a short bicyclo[6.1.0]nonyne (BCN) unit as best suited with regards to conjugation performance and potency. Promising in vitro activity and in vivo pharmacokinetic behavior of optimized Brainshuttle-ASO conjugates, based on a microtubule-associated protein tau (MAPT) targeting oligonucleotide, suggest that such designs have the potential to serve as a blueprint for peripherally delivered ASO-based drugs for the CNS in the future.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: ■ Conclusionmentioning

confidence: 99%

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Diligent Design Enables Antibody-ASO Conjugates with Optimal Pharmacokinetic Properties

Sela,

Mansø,

Siegel

et al. 2023

Bioconjugate Chem.

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“…However, there are many hurdles to overcome before DGAT inhibition becomes a viable therapeutic or preventative option. These include achieving delivery across the blood brain barrier (a challenge which has promising solutions in antibody-mediated delivery 55 ) and correct dosing to avoid toxicity due to inhibition of the key fat storage functions of DGAT enzymes. This latter challenge could perhaps be overcome by exploiting functional redundancy of DGAT1 and DGAT2.…”

Section: Discussionmentioning

confidence: 99%

Triglyceride metabolism controls inflammation andAPOE4-associated disease states in microglia

Stephenson,

Johnson,

Cheng

et al. 2024

Preprint

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Microglia modulate their cell state in response to various stimuli. Changes to cellular lipids often accompany shifts in microglial cell state, but the functional significance of these metabolic changes remains poorly understood. In human induced pluripotent stem cell-derived microglia, we observed that both extrinsic activation (by lipopolysaccharide treatment) and intrinsic triggers (the Alzheimers disease-associated APOE4 genotype) result in accumulation of triglyceride-rich lipid droplets. We demonstrate that lipid droplet accumulation is not simply concomitant with changes in cell state but rather necessary for microglial activation. We discovered that both triglyceride biosynthesis and catabolism are needed for the transcription and secretion of proinflammatory cytokines and chemokines in response to extrinsic stimuli. Additionally, we reveal that triglyceride biosynthesis and catabolism are necessary for the activation-associated phagocytosis of multiple substrates including the disease-associated amyloid-beta peptide. In microglia harboring the Alzheimers disease risk APOE4 genotype, triglyceride-rich lipid droplets accumulate even in the absence of any external stimuli. Inhibiting triglyceride biosynthesis in APOE4 microglia not only modifies the transcription of immune response genes but also attenuates disease-associated transcriptional states. This work establishes that triglyceride metabolism is necessary for microglia to respond to extrinsic activation. In APOE4 microglia, this metabolic process modulates both immune signaling and a disease-associated transcriptional state. Importantly, our work identifies metabolic pathways that can be used to tune microglial immunometabolism in APOE4-associated disease.

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Targeted transport of biotherapeutics at the blood-brain barrier

Moos,

Thomsen,

Burkhart

et al. 2023

Expert Opinion on Drug Delivery

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Targeting Transferrin Receptor to Transport Antisense Oligonucleotides Across the Blood-Brain Barrier

Cited by 10 publications

References 69 publications

Diligent Design Enables Antibody-ASO Conjugates with Optimal Pharmacokinetic Properties

Diligent Design Enables Antibody-ASO Conjugates with Optimal Pharmacokinetic Properties

Triglyceride metabolism controls inflammation andAPOE4-associated disease states in microglia

Targeted transport of biotherapeutics at the blood-brain barrier

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