Neuronally expressed anti-tau scFv prevents tauopathy-induced phenotypes in Drosophila models

Krishnaswamy, Senthilkumar; Huang, Huai-Wei; Marchal, Isabella S.; Ryoo, Hyung Don; Sigurdsson, Einar M.

doi:10.1016/j.nbd.2020.104770

Cited by 18 publications

(10 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our laboratory pioneered targeting pathological tau protein with active and passive immunotherapies for AD and other tauopathies (5,6), which have been validated and extended by other groups over the last several years and now entered clinical trials (1)(2)(3)(4). Antibody-mediated clearance of tau likely involves several mechanisms, which may include: (a) microglia activation and phagocytosis (7)(8)(9)(10), (b) neutralization of tau in the extracellular space (11,12), and (c) intracellular sequestration/degradation of tau within neurons (5,7,(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24). Our group and others have focused on elucidating these mechanisms in various cell culture, ex-vivo, and in-vivo mouse models.…”

Section: Introductionmentioning

confidence: 99%

Dynamics of Internalization and Intracellular Interaction of Tau Antibodies and Human Pathological Tau Protein in a Human Neuron-Like Model

Shamir

Deng

et al. 2020

Front. Neurol.

Self Cite

View full text Add to dashboard Cite

We and others have shown in various in vivo, ex vivo and cell culture models that several tau antibodies interact with pathological tau within neurons. To further clarify this interaction in a dynamic human model, we differentiated SH-SY5Y cells with retinoic acid and BDNF to create a neuron-like model. Therein, tau antibodies were primarily taken up by receptor-mediated endocytosis, and prevented toxicity of human brain-derived paired helical filament-enriched tau (PHF). Subsequently, we monitored in real-time the interaction of antibodies and PHF within endocytic cellular compartments. Cells were pre-treated with fluorescently-tagged PHF and then incubated with tau antibodies, 4E6, 6B2, or non-specific isotype control IgG1 labeled with a pH sensitive dye. The uptake and binding of the efficacious antibody, 4E6, to PHF occurred mainly within the soma, whereas the ineffective antibody, 6B2, and ineffective control IgG1, were visualized via the processes and showed limited colocalization with PHF within this period. In summary, we have developed a neuron-like model that clarifies the early intracellular dynamics of the interaction of tau antibodies with pathological tau, and identifies features associated with efficacy. Since the model is entirely human, it is suitable to verify the therapeutic potential of humanized antibodies prior to extensive clinical trials.

show abstract

Section: Introductionmentioning

confidence: 99%

Dynamics of Internalization and Intracellular Interaction of Tau Antibodies and Human Pathological Tau Protein in a Human Neuron-Like Model

Shamir

Deng

et al. 2020

Front. Neurol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…AAV based scFv expression can help overcome the short half-life of scFv due to their lack of an Fc region. Some studies have shown success in the use of AAV based scFv for Alzheimer’s disease, and Amyotrophic Lateral Sclerosis amongst others ( 61 , 67 , 68 ). In vivo data will be required to determine the antigenicity and bioavailability of scFv.…”

Section: Discussionmentioning

confidence: 99%

Combinations of Single Chain Variable Fragments From HIV Broadly Neutralizing Antibodies Demonstrate High Potency and Breadth

et al. 2021

View full text Add to dashboard Cite

Broadly neutralizing antibodies (bNAbs) are currently being assessed in clinical trials for their ability to prevent HIV infection. Single chain variable fragments (scFv) of bNAbs have advantages over full antibodies as their smaller size permits improved diffusion into mucosal tissues and facilitates vector-driven gene expression. We have previously shown that scFv of bNAbs individually retain significant breadth and potency. Here we tested combinations of five scFv derived from bNAbs CAP256-VRC26.25 (V2-apex), PGT121 (N332-supersite), 3BNC117 (CD4bs), 8ANC195 (gp120-gp41 interface) and 10E8v4 (MPER). Either two or three scFv were combined in equimolar amounts and tested in the TZM-bl neutralization assay against a multiclade panel of 17 viruses. Experimental IC50 and IC80 data were compared to predicted neutralization titers based on single scFv titers using the Loewe additive and the Bliss-Hill model. Like full-sized antibodies, combinations of scFv showed significantly improved potency and breadth compared to single scFv. Combinations of two or three scFv generally followed an independent action model for breadth and potency with no significant synergy or antagonism observed overall although some exceptions were noted. The Loewe model underestimated potency for some dual and triple combinations while the Bliss-Hill model was better at predicting IC80 titers of triple combinations. Given this, we used the Bliss-Hill model to predict the coverage of scFv against a 45-virus panel at concentrations that correlated with protection in the AMP trials. Using IC80 titers and concentrations of 1μg/mL, there was 93% coverage for one dual scFv combination (3BNC117+10E8v4), and 96% coverage for two of the triple combinations (CAP256.25+3BNC117+10E8v4 and PGT121+3BNC117+10E8v4). Combinations of scFv, therefore, show significantly improved breadth and potency over individual scFv and given their size advantage, have potential for use in passive immunization.

show abstract

“…The affinity could also be improved by re-cloning and re-selection of clones or by conversion of sdAbs into multivalent formats with higher avidity ( 4 , 42 ). Though antibody affinity can be significant for its potential for immunotherapy, most interestingly however, it was found that low, rather than high antibody affinity has been reported to essential for the passive antibody therapy of a drosophila based model of Alzheimer’s disease ( 43 , 44 ). This was interpreted on the grounds that the low affinity anti-tau antibody may loosen up intracellular tau aggregates allowing better access of lysosomal degrading enzymes, while high affinity antibody may make these aggregates more compact and therefore more difficult to degrade.…”

Section: Discussionmentioning

confidence: 99%

Selection of a Single Domain Antibody, Specific for an HLA-Bound Epitope of the Mycobacterial Ag85B Antigen

et al. 2020

View full text Add to dashboard Cite

T cells recognizing epitopes on the surface of mycobacteria-infected macrophages can impart protection, but with associated risk for reactivation to lung pathology. We aimed to identify antibodies specific to such epitopes, which carry potentials for development toward novel therapeutic constructs. Since epitopes presented in the context of major histocompatibility complex alleles are rarely recognized by naturally produced antibodies, we used a phage display library for the identification of monoclonal human single domain antibody producing clones. The selected 2C clone displayed T cell receptor-like recognition of an HLA-A*0201 bound 199 KLVANNTRL 207 peptide from the Ag85B antigen, which is known to be an immunodominant epitope for human T cells. The specificity of the selected domain antibody was demonstrated by solid phase immunoassay and by immunofluorescent surface staining of peptide loaded cells of the T2 cell line. The antibody affinity binding was determined by biolayer interferometry. Our results validated the used technologies as suitable for the generation of antibodies against epitopes on the surface of Mycobacterium tuberculosis infected cells. The potential approaches forward the development of antibody in immunotherapy of tuberculosis have been outlined in the discussion.

show abstract

Neuronally expressed anti-tau scFv prevents tauopathy-induced phenotypes in Drosophila models

Cited by 18 publications

References 47 publications

Dynamics of Internalization and Intracellular Interaction of Tau Antibodies and Human Pathological Tau Protein in a Human Neuron-Like Model

Dynamics of Internalization and Intracellular Interaction of Tau Antibodies and Human Pathological Tau Protein in a Human Neuron-Like Model

Combinations of Single Chain Variable Fragments From HIV Broadly Neutralizing Antibodies Demonstrate High Potency and Breadth

Selection of a Single Domain Antibody, Specific for an HLA-Bound Epitope of the Mycobacterial Ag85B Antigen

Contact Info

Product

Resources

About