Local delivery of stabilized chondroitinase ABC degrades chondroitin sulfate proteoglycans in stroke-injured rat brains

Hettiaratchi, Marian H.; O’Meara, Matthew J.; Teal, Carter J.; Payne, Samantha L.; Pickering, Andrew; Shoichet, Molly S.

doi:10.1016/j.jconrel.2019.01.033

Cited by 45 publications

(43 citation statements)

References 47 publications

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“…To achieve sustained release of the ChABC-SH3 designs using an injectable, affinity-controlled hydrogel delivery system (3,27,28), we modified 5% (w/v) thiolated MC hydrogels cross-linked with PEG bis-maleimide with (or without) SH3-binding peptides (100:1 molar ratio of SH3-binding peptide to fusion protein) and separately incorporated ChABC-37-SH3 and ChABC-SH3 therein (Fig. 5A).…”

Section: Affinity-controlled Release Of Chabc-sh3mentioning

confidence: 99%

“…While this affinity-based delivery strategy only resulted in release of ~62% of the loaded ChABC-37-SH3 over 7 days in vitro, we expect full protein release to be achieved in vivo due to gradual dissolution of the hydrogel. A prolonged effect of a less stable form of ChABC-SH3, delivered using this strategy, was observed for up to 2 and 4 weeks in the spinal cord and brain, respectively (3,28). To facilitate complete protein release, we are further engineering the MC delivery vehicle to expedite degradation and resorption in vivo.…”

Section: Chondroitin Sulfate a Dermatan Sulfatementioning

confidence: 99%

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Reengineering biocatalysts: Computational redesign of chondroitinase ABC improves efficacy and stability

Hettiaratchi

O’Meara

et al. 2020

Sci. Adv.

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Maintaining biocatalyst stability and activity is a critical challenge. Chondroitinase ABC (ChABC) has shown promise in central nervous system (CNS) regeneration, yet its therapeutic utility is severely limited by instability. We computationally reengineered ChABC by introducing 37, 55, and 92 amino acid changes using consensus design and forcefield-based optimization. All mutants were more stable than wild-type ChABC with increased aggregation temperatures between 4° and 8°C. Only ChABC with 37 mutations (ChABC-37) was more active and had a 6.5 times greater half-life than wild-type ChABC, increasing to 106 hours (4.4 days) from only 16.8 hours. ChABC-37, expressed as a fusion protein with Src homology 3 (ChABC-37-SH3), was active for 7 days when released from a hydrogel modified with SH3-binding peptides. This study demonstrates the broad opportunity to improve biocatalysts through computational engineering and sets the stage for future testing of this substantially improved protein in the treatment of debilitating CNS injuries.

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Section: Affinity-controlled Release Of Chabc-sh3mentioning

confidence: 99%

Section: Chondroitin Sulfate a Dermatan Sulfatementioning

confidence: 99%

Reengineering biocatalysts: Computational redesign of chondroitinase ABC improves efficacy and stability

Hettiaratchi

O’Meara

et al. 2020

Sci. Adv.

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“…These findings combined with our findings of increased percentage of Arg1+ cells, suggests that microporous hydrogels formed using ~80 µm HMPs have pore sizes that can spatially confine the microglia and lower M1 polarization. Chondroitin sulfate proteoglycans (CSPGs) have been linked to decreased regenerative potential in the CNS 28,44,45 .…”

Section: Microglia Up-regulate Pro-repair Marker Arg1 In Ha-mapmentioning

confidence: 99%

“…Recently others [21][22][23] and we 24,25 have published a number of publications injecting hydrogels into the stroke cavity. These studies have demonstrated that the stroke core can accept a significant gel injection volume without damage and that these gels can vary widely in composition ranging from hydrogels formed from decellularized native tissue 26 , to peptide derived hydrogels 27 , to synthetic hydrogels 28 . Although biomaterial strategies for brain repair have investigated astrocytes after stroke in the context of the glial scar, no biomaterial has been described that can modulate the astrocyte phenotype from inflammatory to pro-regenerative.…”

Section: Introductionmentioning

confidence: 99%

Hyaluronic acid particle hydrogels decrease cerebral atrophy and promote pro-reparative astrocyte/axonal infiltration in the core after ischemic stroke

Sideris

Chen

et al. 2019

Preprint

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The death rate due to stroke is decreasing, resulting in more individuals living with stroke related disabilities. Following stroke, dying cells contribute to the large influx of highly reactive astrocytes and pro-inflammatory microglia that release cytokines and lead to a cytotoxic environment that causes further brain damage and prevents endogenous repair. Paradoxically, these same cells also activate pro-repair mechanisms that contribute to endogenous repair and brain plasticity. Here, we show that the direct injection of a hyaluronic acid based microporous annealed particle (HA-MAP) hydrogel into the stroke core reduces the percent of highly reactive astrocytes and increases the percent of alternatively activated microglia in and around the lesion. Further, we show that HA-MAP hydrogel promotes reparative astrocyte infiltration into the lesion, which directly coincides with axonal penetration into the lesion. Additionally, HA-MAP injection decreases cerebral atrophy and preserves nigrostriatal bundles after stroke. This work shows that the injection of a porous scaffold into the stroke core can lead to clinically relevant decrease in cerebral atrophy and modulates the phenotype of astrocytes and microglia towards a pro-repair phenotype.

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“…However, the effects of exogenous NP/SCs hinder the restoration of brain function, primarily due to the low survival rate of grafted cells in host tissues [7,8], which may be caused by mechanical damage, acute inflammation, immunological rejection, or a lack of trophic signals [9]. After brain injury, glial cells are activated and proliferate rapidly to become reactive glial cells in the injured cortex, forming a glial scar that prevents neuronal growth inside the brain [10,11]. In recent years, neuronal reprogramming by the conversion of non-neuronal cells into functional neurons has represented a substantial improvement in the field of repairing injured or diseased brains [12,13].…”

Section: Introductionmentioning

confidence: 99%

Neurogenic Niche Conversion Strategy Induces Migration and Functional Neuronal Differentiation of Neural Precursor Cells Following Brain Injury

Wang

Zheng

et al. 2020

Stem Cells and Development

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Glial scars formed after brain injuries provide permissive cues for endogenous neural precursor/stem cells (eNP/ SCs) to undergo astrogenesis rather than neurogenesis. Following brain injury, eNP/SCs from the subventricular zone leave their niche, migrate to the injured cortex, and differentiate into reactive astrocytes that contribute to glial scar formation. In vivo neuronal reprogramming, directly converting non-neuronal cells such as reactive astrocytes or NG2 glia into neurons, has greatly improved brain injury repair strategies. However, reprogramming carries a high risk of future clinical applications such as tumorigenicity, involving virus. In this study, we constructed a neural matrix to alter the adverse niche at the injured cortex, enabling eNP/SCs to differentiate into functional neurons. We found that the neural matrix functioned as a ''glial trap'' that largely concentrated and limited reactive astrocytes to the core of the lesion area, thus altering the adverse niche. The eNP/SCs migrated toward the injured cortex and differentiated into functional neurons. In addition, regenerated neurites extended across the boundary of the injured cortex. Mice treated with the neural matrix demonstrated significant behavioral recovery. For the first time, we induced eNP/SC-derived functional neurons in the cortex after brain injury without the use of viruses, microRNAs, or small molecules. Our novel strategy of applying this ''glial trap'' to obtain functional neurons in the injured cortex may provide a safer and more natural therapeutic alternative to reprogramming in future clinical applications.

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Local delivery of stabilized chondroitinase ABC degrades chondroitin sulfate proteoglycans in stroke-injured rat brains

Cited by 45 publications

References 47 publications

Reengineering biocatalysts: Computational redesign of chondroitinase ABC improves efficacy and stability

Reengineering biocatalysts: Computational redesign of chondroitinase ABC improves efficacy and stability

Hyaluronic acid particle hydrogels decrease cerebral atrophy and promote pro-reparative astrocyte/axonal infiltration in the core after ischemic stroke

Neurogenic Niche Conversion Strategy Induces Migration and Functional Neuronal Differentiation of Neural Precursor Cells Following Brain Injury

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