Intrabody-mediated diverting of HP1β to the cytoplasm induces co-aggregation of H3–H4 histones and lamin-B receptor

Cardinale, Alessio; Filesi, Ilaria; Singh, Prim B.; Biocca, Silvia

doi:10.1016/j.yexcr.2015.09.006

Cited by 6 publications

(4 citation statements)

References 68 publications

(95 reference statements)

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“…Previous studies have documented the efficacy of PEST-fusion intrabodies against pathogenic proteins such as huntingtin 46 and α-synuclein 51 . It was also shown that intrabodies can disrupt the subcellular distribution of target proteins, including by inhibition of nuclear−cytoplasmic translocation 52 , 53 . Although cytoplasmic nonaggregated TDP-43 (TDP-43 mNLS ) was not degraded by 3B12A scFv in cultured cells (Fig.…”

Section: Discussionmentioning

confidence: 99%

Elimination of TDP-43 inclusions linked to amyotrophic lateral sclerosis by a misfolding-specific intrabody with dual proteolytic signals

Tamaki

Shodai

Morimura

et al. 2018

Sci Rep

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Aggregation of TAR DNA-binding protein of 43 kDa (TDP-43) is implicated in the pathogenesis of sporadic and certain familial forms of amyotrophic lateral sclerosis (ALS), suggesting elimination of TDP-43 aggregates as a possible therapeutic strategy. Here we generated and investigated a single-chain variable fragment (scFv) derived from the 3B12A monoclonal antibody (MAb) that recognises D247 of the TDP-43 nuclear export signal, an epitope masked in the physiological state. In transfected HEK293A cells, 3B12A scFv recapitulated the affinity of the full-length MAb to mislocalised TDP-43 with a defective nuclear localising signal and to a TDP-43 inclusion mimic with cysteine-to-serine substitution at RRM1. Moreover, 3B12A scFv accelerated proteasome-mediated degradation of aggregated TDP-43, likely due to an endogenous PEST-like proteolytic signal sequence in the VH domain CDR2 region. Addition of the chaperone-mediated autophagy (CMA)-related signal to 3B12A scFv induced HSP70 transcription, further enhancing TDP-43 aggregate clearance and cell viability. The 3B12A scFv also reduced TDP-43 aggregates in embryonic mouse brain following in utero electroporation while causing no overt postnatal brain pathology or developmental anomalies. These results suggest that a misfolding-specific intrabody prone to synergistic proteolysis by proteasomal and autophagic pathways is a promising strategy for mitigation of TDP-43 proteinopathy in ALS.

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Section: Discussionmentioning

confidence: 99%

Elimination of TDP-43 inclusions linked to amyotrophic lateral sclerosis by a misfolding-specific intrabody with dual proteolytic signals

Tamaki

Shodai

Morimura

et al. 2018

Sci Rep

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“…Protein relocation is an approach to interfere with target protein function through trapping it with binder molecule and thereby preventing the target protein transport to a defined cellular compartment. The feasibility of such strategy was demonstrated with intracellular antibodies, which inhibited heterochromatin protein 1β traffic to the nucleus ( Cardinale et al, 2015 ), resulting in altered nuclear morphology and apoptosis. Another group reported intracellular antibody binding to Sec61 and prevention of its transport from the ER toward endosomes ( Zehner et al, 2015 ).…”

Section: Strategies For Intracellular Targeting Of Antibodies Their mentioning

confidence: 99%

Targeted Intracellular Delivery of Antibodies: The State of the Art

et al. 2018

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A dominant area of antibody research is the extension of the use of this mighty experimental and therapeutic tool for the specific detection of molecules for diagnostics, visualization, and activity blocking. Despite the ability to raise antibodies against different proteins, numerous applications of antibodies in basic research fields, clinical practice, and biotechnology are restricted to permeabilized cells or extracellular antigens, such as membrane or secreted proteins. With the exception of small groups of autoantibodies, natural antibodies to intracellular targets cannot be used within living cells. This excludes the scope of a major class of intracellular targets, including some infamous cancer-associated molecules. Some of these targets are still not druggable via small molecules because of large flat contact areas and the absence of deep hydrophobic pockets in which small molecules can insert and perturb their activity. Thus, the development of technologies for the targeted intracellular delivery of antibodies, their fragments, or antibody-like molecules is extremely important. Various strategies for intracellular targeting of antibodies via protein-transduction domains or their mimics, liposomes, polymer vesicles, and viral envelopes, are reviewed in this article. The pitfalls, challenges, and perspectives of these technologies are discussed.

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“…The generation of the different types of intrabodies also requires very different amounts of effort. Although aggregating intrabodies have also been reported to cause a phenotype [12] , cytosolic intrabodies, ideally i. need to fold correctly in the cytosol in order to be functional and ii. need to bind to a selected epitope in a way allowing neutralization of the target function.…”

Section: Different Types Of Intrabodiesmentioning

confidence: 99%

“…Inhibition of one of the several functions of a target protein can furthermore be achieved by the ability of intrabodies fused to a signal sequence to relocate proteins, as has been demonstrated by the knockdown of Sec61, which was relocated intracellularly by an ER-intrabody to inhibit its function in the endosomes without disrupting its vital function for protein biosynthesis in the ER [53] . Another study reported the re-localization of a target protein to prevent its function in the nucleus [12] . A similar approach allowed to block the pathogenic polymerization of Z α1-antitrypsin while maintaining its antiproteinase activity without which patients would develop a lung disease [37] .…”

Section: Intrabodies For Therapy?mentioning

confidence: 99%

Antibodies inside of a cell can change its outside: Can intrabodies provide a new therapeutic paradigm?

Marschall

Dübel

2016

Computational and Structural Biotechnology Journal

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Challenges posed by complex diseases such as cancer, chronic viral infections, neurodegenerative disorders and many others have forced researchers to think beyond classic small molecule drugs, exploring new therapeutic strategies such as therapy with RNAi, CRISPR/Cas9 or antibody therapies as single or as combination therapies with existing drugs. While classic antibody therapies based on parenteral application can only reach extracellular targets, intracellular application of antibodies could provide specific advantages but is so far little recognized in translational research. Intrabodies allow high specificity and targeting of splice variants or post translational modifications. At the same time off target effects can be minimized by thorough biochemical characterization. Knockdown of cellular proteins by intrabodies has been reported for a significant number of disease-relevant targets, including ErbB-2, EGFR, VEGFR-2, Metalloproteinase MMP2 and MMP9, β-amyloid protein, α-synuclein, HIV gp120, HCV core and many others. This review outlines the recent advances in ER intrabody technology and their potential use in therapy.

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Intrabody-mediated diverting of HP1β to the cytoplasm induces co-aggregation of H3–H4 histones and lamin-B receptor

Cited by 6 publications

References 68 publications

Elimination of TDP-43 inclusions linked to amyotrophic lateral sclerosis by a misfolding-specific intrabody with dual proteolytic signals

Elimination of TDP-43 inclusions linked to amyotrophic lateral sclerosis by a misfolding-specific intrabody with dual proteolytic signals

Targeted Intracellular Delivery of Antibodies: The State of the Art

Antibodies inside of a cell can change its outside: Can intrabodies provide a new therapeutic paradigm?

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