Machine-Assisted Discovery of Chondroitinase ABC Complexes toward Sustained Neural Regeneration.
Machine-Assisted Discovery of Chondroitinase ABC Complexes toward Sustained Neural Regeneration.
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DOI：
10.1002/adhm.202102101
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发表时间：
2022-05
影响因子：
10
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中科院分区：
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在许多导致胶质瘢痕形成的分子中，硫酸软骨素蛋白多糖（CSPGs）被认为是神经元再生的有效抑制剂。软骨素酶ABC （ChABC）是一种细菌裂解酶，已知可降解CSPGs的糖胺聚糖（GAG）侧链并促进组织再生。然而，ChABC是热不稳定的，在37°C的稀释条件下，几个小时内就会失去所有活性。为了克服这一限制，我们报告了一组定制的随机共聚物的发现，这些共聚物在生理温度下使ChABC复合并稳定。共聚物设计基于共聚物的链长和组成，使用主动机器学习范式进行识别，其中包括迭代共聚物合成，共聚物络合时的ChABC热稳定性测试，高斯过程回归建模和贝叶斯优化。共聚物由自动PET-RAFT合成，在37℃下放置24小时后，通过酶活性（REA）来评估ChABC的热稳定性。在三次主动学习迭代中，我们证明了REA的显著改进，同时确定了异常高性能的共聚物。最值得注意的是，一种设计的共聚物即使在一周后也能将剩余ChABC活性提高近30%，并且明显优于其他常见的ChABC稳定方法。总之，这些结果强调了一条有希望的持续组织再生途径。ChABC聚合物酶复合物。设计的合成共聚物在变性环境中保持了ChABC的功能。
Among the many molecules that contribute to glial scarring, chondroitin sulfate proteoglycans (CSPGs) are known to be potent inhibitors of neuronal regeneration. Chondroitinase ABC (ChABC), a bacterial lyase, is known to degrade the glycosaminoglycan (GAG) side chains of CSPGs and promote tissue regeneration. However, ChABC is thermally unstable and loses all activity within a few hours at 37°C under dilute conditions. To overcome this limitation, we report the discovery of a diverse set of tailor-made random copolymers that complex and stabilize ChABC at physiological temperature. The copolymer designs, which are based on chain length and composition of the copolymers, were identified using an active machine learning paradigm, which involves iterative copolymer synthesis, testing for ChABC thermostability upon copolymer complexation, Gaussian Process Regression modeling, and Bayesian optimization. Copolymers are synthesized by automated PET-RAFT and thermostability of ChABC is assessed by retained enzyme activity (REA) after 24 hrs at 37°C. We demonstrated significant improvements in REA in three iterations of active learning while identifying exceptionally high-performing copolymers. Most remarkably, one designed copolymer promoted residual ChABC activity near 30%, even after one week and notably outperformed other common stabilization methods for ChABC. Together, these results highlight a promising pathway towards sustained tissue regeneration. ChABC Polymer Enzyme Complex. Designed synthetic copolymers preserve ChABC function in denaturing environments.