Engineering Brain Parasites for Intracellular Delivery of Therapeutic Proteins
Engineering Brain Parasites for Intracellular Delivery of Therapeutic Proteins
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DOI：
10.1101/481192
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发表时间：
2018-12
期刊：
影响因子：
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通讯作者：
Shahar Bracha;Karoliina Hassi;P. Ross;S. Cobb;L. Sheiner;O. Rechavi
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文献类型：
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作者：
Shahar Bracha;Karoliina Hassi;P. Ross;S. Cobb;L. Sheiner;O. Rechavi
蛋白质疗法具有缓解许多神经系统疾病的潜力;然而，中枢神经系统（CNS）的递送机制是有限的，并且细胞内递送构成了额外的障碍。为了应对这些挑战，我们利用原生生物寄生虫弓形虫，它可以迁移到中枢神经系统并将蛋白质分泌到细胞中。使用融合蛋白的方法，我们改造了T。分泌治疗人类神经系统疾病的蛋白质。我们测试了两个分泌系统，产生了定位于T。的分泌细胞器，并评估其在各种哺乳动物细胞，包括神经元的细胞内靶向。我们证明了T.表达融合到Rett综合征蛋白MeCP 2的GRA 16的弓形虫递送模拟内源性MeCP 2的融合蛋白，结合神经元中的异染色质DNA。这表明T.弓形虫作为治疗性蛋白质载体，其可以提供瞬时或慢性的原位合成并将细胞内蛋白质递送至CNS。
Protein therapy has the potential to alleviate many neurological diseases; however, delivery mechanisms for the central nervous system (CNS) are limited, and intracellular delivery poses additional hurdles. To address these challenges, we harnessed the protist parasite Toxoplasma gondii, which can migrate into the CNS and secrete proteins into cells. Using a fusion protein approach, we engineered T. gondii to secrete therapeutic proteins for human neurological disorders. We tested two secretion systems, generated fusion proteins that localized to T. gondii’s secretory organelles and assessed their intracellular targeting in various mammalian cells including neurons. We show that T. gondii expressing GRA16 fused to the Rett syndrome protein MeCP2 deliver a fusion protein that mimics the endogenous MeCP2, binding heterochromatic DNA in neurons. This demonstrates the potential of T. gondii as a therapeutic protein vector, which could provide either transient or chronic, in situ synthesis and delivery of intracellular proteins to the CNS.